PROCEEDINGS OF SUGARCANE 2023

199 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry CO-PRODUCTS Figure 1 Three-dimensional response surface and contour plots of bacterial growth (a and b), biosurfactant concentration (c and d) and lignin concentration (e and f) s howing the effect of mixed sugarcane filter cake and yeast sludge.

200 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry CO-PRODUCTS 3.3 Surface activity of biosurfactant in cell-free broth Assessment of the contact angle reduction of cell-free broth with and without lignin showed contact angle reductions lower than 90 degrees on a glass slide, stainless steel, T. procumbens, B. oleracea, and sugarcane leaf, while higher than 90 degrees on a parafilm (Figure 2). Consequently, the application of the produced biosurfactant can be applied as bioactive enhancement in agriculture and bio cleaning in factories. The low contact angle reduction with parafilm might come from the low hydrophobicity of biosurfactant resulting from the use of non-oil substrates. Moreover, lignin in cell-free broth has no negative effect on contact angle reduction by biosurfactant. Therefore, cell-free broth can be used after biosurfactant production without the lignin removal process. The Surface tension reduction of cell-free broth wassimilarto that of commercialrhamnolipid but lessthan that of commercialsophorolipid and tween80. Figure 2 Contact angle reduction on a glass slide, stainless steel, parafilm, T. procumbens, B. oleracea, and sugarcane leaf of a produced biosurfactant in cell-free broth with and without lignin 4. CONCLUSIONS Our research demonstrates clearly that Brevibacterium casei NK8 produced a high amount of biosurfactant during submerged fermentation on substrates derived from mixing agro-industrial wastes. Notably, the biosurfactant was enabled its activities over various materials and plants. The findings suggest that the produced biosurfactant can readily be implemented for fulfilling the enormous demand of surfactant in cleaning and agriculture. 5. ACKNOWLEDGEMENT The first author and the corresponding authorthank the Faculty ofAgricultureNaturalResources andEnvironment at Naresuan University for providing funding to carry out the present research through capacity-building projects. 6. REFERENCES [1] Vandenberghe, L.P.S., Valladares-Diestra, K.K., Bittencourt, G.A., Torres, L.Z., Vieira, S., Karp, S.G., Sydney, E.B., de Carvalho, J.C., Soccol, V.T. and Soccol, C.R. (2022). Beyond sugar and ethanol: The future of sugarcane biorefineries in Brazil. Renewable and Sustainable Energy Reviews, 167, pp. 112721. [2] Cardoso, T.F., Watanabe, M.D., Souza, A., Chagas, M.F., Cavalett, O., Morais, E.R., Nogueira, L.A., Leal, M.R.L., Braunbeck, O.A., Cortez, L.A. and Bonomi, A. (2018). Economic, environmental, and social impacts of different sugarcane production systems. Biofuels, Bioproducts and Biorefining, 12(1), pp. 68-82. [3] Formann, S., Hahn, A., Janke, L., Stinner, W., Sträuber, H., Logroño, W. and Nikolausz, M. (2020). Beyond sugar and ethanol production: value generation opportunities through sugarcane residues. Frontiers in Energy Research, 8, pp. 579577. [4] Joy, S., Rahman, P.K., Khare, S.K., Soni, S.R. and Sharma, S. (2019). Statistical and sequential (fill-and-draw) approach to enhance rhamnolipid production using industrial lignocellulosic hydrolysate C6 stream from Achromobacter sp. (PS1). Bioresource technology, 288, pp. 121494. [5] Jimoh, A.A. and Lin, J. (2019). Biosurfactant: A new frontier for greener technology and environmental sustainability. Ecotoxicology and Environmental safety, 184, pp.109607. [6] Makkar, R.S., Cameotra, S.S. and Banat, I.M. (2011). Advances in utilization of renewable substrates for

201 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry CO-PRODUCTS biosurfactant production. AMB express, 1, pp.1-19. [7] Khondee, N., Ruamyat, N., Luepromchai, E., Sikhao, K., & Hawangchu, Y. (2022). Bioconversion of lignocellulosic wastes to zwitterionic biosurfactants by an alkaliphilic bacterium: Process development and product characterization. Biomass and Bioenergy, 165, pp. 106568. [8] Romaní, A., Tomaz, P.D., Garrote, G., Teixeira, J.A. and Domingues, L. (2016). Combined alkali and hydrothermal pretreatments for oat straw valorization within a biorefinery concept. Bioresource technology, 220, pp. 323-332. [9] Wang, G. and Chen, H. (2013). Fractionation of alkali-extracted lignin from steam-exploded stalk by gradient acid precipitation. Separation and Purification Technology, 105, pp. 98-105. [10] Khondee, N., Tathong, S., Pinyakong, O., Müller, R., Soonglerdsongpha, S., Ruangchainikom, C., Tongcumpou, C. and Luepromchai, E. (2015). Lipopeptide biosurfactant production by chitosan-immobilized Bacillus sp. GY19 and their recovery by foam fractionation. Biochemical Engineering Journal, 93, pp. 47-54. [11] Xia, W.J., Luo, Z.B., Dong, H.P., Yu, L., Cui, Q.F. and Bi, Y.Q. (2012). Synthesis, characterization, and oil recovery application of biosurfactant produced by indigenous Pseudomonas aeruginosa WJ-1 using waste vegetable oils. Applied biochemistry and biotechnology, 166, pp. 1148-1166. [12] Jain, R.M., Mody, K., Joshi, N., Mishra, A. and Jha, B. (2013). Effect of unconventional carbon sources on biosurfactant production and its application in bioremediation. International journal of biological macromolecules, 62, pp. 52-58.

202 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry CO-PRODUCTS Phatthilakorn Chamnanpuen and Kittiwut Kasemwong* Nano Agricultural Chemistry and Processing Research Team (ACP), National Nanotechnology Center (NANOTEC), National Science and Technology Development Agency (NSTDA), Pathumthani, 12120, Thailand *Correspondence to: Nano Agricultural Chemistry and Processing Research Team (ACP), National Nanotechnology Center (NANOTEC), National Science and Technology Development Agency (NSTDA), Pathumthani, 12120, Thailand. E-mail: [email protected] ABSTRACT: Policosanol is a naturalmixture oflong-chain alcoholswith carbon backbonesranging from24 to 34 carbons. These compounds can be isolated from waste (sugarcane filter cake) during sugar production that were a high value compounds to either food, cosmetic or pharmaceutical industries in the present. Up to date, there are several reports demonstrated that policosanol have been significant effect on many biological activities, especially, reduction of low-density lipoproteins cholesterol (LDL-C) and increasing biosynthesis of high-density lipoproteins cholesterol (HDL-C). However, policosanol have been known as a commercial dietary supplement product claimed as a natural lipid -lowering agent, there are still conflicting the finding about the efficacy of policosanol or its low bioavailability of policosanol after oral administration. Hence, there is a need to enhance its bioavailability, and nanoemulsion formulation is the interesting one. The aim of this study is to enhance the oral bioavailability of sugarcane wax policosanol. The sugarcane wax policosanol was applied in nanoemulsion formulation using low-energy emulsification method (phase inversion composition) to achieved a highly stable nanoemulsion with a minimum particle size. There were shown the suitable proportion of sugarcane wax policosanol, oil, emulsifier, and co-emulsifier can give a minimum particle size (less than 50 nm), it may be possible delivery the policosanol through intestinal cells. These data can be an advantage as guidance in development of a natural cholesterol-lowering drug or used as food supplements. Keyword: Sugarcane wax, Policosanol, Nanoemulsion, Oral bioavailability P-021 Oral Bioavailability Enhancement of Sugarcane Wax Policosanol Nanoemulsion

203 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry CO-PRODUCTS Yupadee Paopun1 *, Piyanan Thanomchat¹ and Donruedee Toyen² 1 Scientific Equipment and Research Division, Kasetsart University Research and Development Institute, Bangkok, 10900, Thailand ²Department of Materials Science, Faculty of Science, Kasetsart University, Bangkok, 10900, Thailand *Correspondence to: Scientific Equipment and Research Division, Kasetsart University Research and Development Institute, 50 Ngamwongwan Road Ladyao Chatuchak, Bangkok, 10900, Thailand. [email protected] ABSTRACT: Sugarcane is a plant that accumulates silica in the root, stem, leaf, and leaf sheath. The leaf sheath is a part of the agricultural waste of post-harvest in the field. The agriculturist prefers to burn them in their fields which causes air pollution problems. Therefore, it is important to find a solution to reduce pollution in the air. In addition, it can increase the value of the agricultural waste for agriculturists. The aim of this study was to investigate silica contents in dry, raw leaf sheaths of four most popular sugarcane cultivars grown in Thailand. They included KPS01-12, KK3, UT84-12 and LK92-11. Sugarcane leaf sheath surfaces were examined with field-emission scanning electron microscopy (FE-SEM) equipped with energy-dispersive X-ray spectroscopy (EDS). The SiO2 contents were also determined with energy-dispersive X-ray fluorescence (EDXRF). SEM imaging results show that the distribution of Si on the abaxial is higher than on the adaxial of the sugarcane leaf sheath surface. SEM/EDS and XRF show that each cultivar contains different amount of Si. The SEM/EDS mapping show that the KPS01-12 had the highest Si of 12.95±0.95 wt% on the lower epidermis of the leaf sheath surface and 0.66±0.14 wt% on the upper epidermis of the leaf sheath. The XRF analysis of 1 gram of leaf sheath powder shows that the KK3 and KPS01-12 contain the highest Si of 4.10±0.27 and 4.02±0.47 wt%, respectively. This study demonstrates that sugarcane leaf sheath, an agricultural waste, contains bio-silica which could be extracted to be used for other benefits and applications such as in composite materials for fertilizers, rubber products, paints, cosmetics, and others. Keywords: Sugarcane, Silica, Abaxial, Adaxial, Epidermis, Leaf sheath P-026 Determination of Bio Silica in Leaf Sheath of Sugarcane

204 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry CO-PRODUCTS 1. INTRODUCTION Sugarcane (Sasscharum officinaram L.) was classified as a family of Poaceae [1]. Sugarcane is an important Thai economic plant. There are approximately 1.75 million hectares of sugarcane plantations in Thailand [2]. This plant accumulates silica in the root, stem, leaf and leaf sheath. Silica appeared in epidermis cells more than chlorenchyma cells [3]. The leaf sheath of sugarcane is a part of the agricultural waste of post-harvest left in the field. The agriculturists usually burn them in their fields due to convenience and low cost which causes air pollution and health problems. Sornpoon et al. (2014) reported that the sugarcane field was a main source of air pollution. Poltam et al. (2018) reported that the burning of sugarcane leaves in the fields created pollution that causes allergies for humans and danger to other living organisms. Moreover, it also increases the amount of carbon dioxide and heat in the air, which causes global warming. In order to help with these issues, it is important to find an alternative solution to handle the waste. Sugarcane is one of the plants in the grass family, which accumulated silicon in several parts such as leaves, shoots, stalks and roots. Sahebi et al. (2015) reported that silicon is an important element in plants. Grasses accumulated silica in the leaves about 2-5% of dry leaf mass in order to fight against insects and herbivores [7]. Si uptake in plants is in silicic acid [Si (OH)4] form which could increase the strength of the cell wall [8]. Accumulated silica could also be in the form of amorphous silica [9]. The rigid structure of the cell made plants resistant to diseases and insects [8]. Prasad (2019) proposed that silicon could protect plants against herbivores, insects and diseases. Vandegeer et al. (2021) reported that silicon on stomata could protect Festuca arundinacea Schreb. cv. Fortuna from water stress. Matichenkov and Calvert (2002) found that an important function of silicon was promoted against abiotic and biotic stress in the plant such as against sugarcane rust, sugarcane ringspot, and leaf freckle. Kaufman et al. (1985) found that silica in the grass leaf was accumulated in silica cells, long epidermal cells and bulliform cells of rice. Paopun et al. (2021) reported that the leaf surface of the Suphanburi 50 cultivar of Thai sugarcane structure was composed of epidermal cells, guard cells, subsidiary cells, cork cells, and silica cells. EDS indicated that the leaf surface contained the highest Si in the apical part and the lower epidermis of the leaf sheath contained 8.96-16.08wt% Si. The leaf sheath is the part that connects between leaf blade with the stem. The function of the leaf sheath is to support and protect the young stem from the environment, especially from wind storms that may cause breaking. The anatomical characteristics of leaf sheath component with upper epidermis, air space, stacked bundle, and lower epidermis. Collenchyma bundles appear under the upper epidermis. The lower epidermis consists of sclerenchyma cells which are long cells and thick-walled cells. The outer epidermis cells with a wavy wall containing silica and connector cells, stomata, and hairs [15]. Paopun et al. (2022) reported that the silica accumulated in the sugarcane leaf of the UT84-12 cultivar was 6.34 wt% when analyzed by EDXRF. However, SEM/EDS showed that the silicon on the abaxial leaf surface was 19.77 wt%. Alves et al. (2017) found that the bio-silica extracted from sugarcane waste ash was in a mesoporous structure which could be made into a catalyst support and an adsorbent material. 2. MATERIALS AND METHODS Sugarcane leaf sheath material The sugarcane leaf sheaths were collected from the field in Song Phi Nong District, Suphanburi Province, Thailand. They were KPS01-12, KK3, UT84-12, and LK92-11 cultivars of Thai sugarcane. The samples were examined by an energy-dispersive X-ray fluorescence spectrometer (EDXRF), and a field emission scanning electron microscope & equipped with an energy-dispersive X-ray spectrometer (FE-SEM&EDS). EDXRF analysis The sugarcane leaf sheath was ground by a cutting mill (Retsch: SM100). One gram of sample powder was put into a sample cup. The sample was analyzed by the EDXRF (Rigaku: NEXDE), operated at 60kV for 12 min. A silicon drift detector (SDD) is an X-ray detector. FE-SEM analysis of Sugarcane leaf sheath The sugarcane leaf sheath samples were cleaned with distilled water and dried in a hot air oven at 45°C for 8 hrs. They were cut into 1x1 cm2 and pasted on an aluminum stub with carbon tape and kept in a desiccator. The samples were coated with a carbon thin film by carbon coater (Quorum: Q150R). The prepared samples were examined by a FE-SEM (Hitachi: SU8020) operated at 15 kV and elemental analysis was carried outs by EDS (EDAX: Apollo X) [18]. The quantitative analysis of elements and silicon distribution were determined using an area mode of energy-dispersive X-ray spectrometer, ten areas of the leaf sheath, about 400x510µm2 were analyzed by EDS. 3. RESULTS AND DISCUSSION Surfaces of four cultivars ofsugarcane leafsheath were examined by SEM, including KPS01-12, KK3, UT84-12, and LK92-11. The leaf sheath surface has to components: the upper epidermis (adaxial) and lower epidermis layers

205 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry CO-PRODUCTS (abaxial). Each of the epidermis layers has different structural characters. The upper epidermis layer appeared the epidermal cells and stomata while, the lower epidermis layer has the epidermal cells, stomata, silica cells, unicellular trichome, and bicellular trichome (Figure 1). This is in good agreement with a study by Moore (1987) that the upper epidermis layer of sugarcane had large cells and uniformly rectangular cells and a few stomata. On the contrary, the lower epidermis layer composed of narrow and long epidermal cells, stomata, silica cells, cork cells and trichomes (hair) (Figure 1, Figure 2). Figures 1 A, C, E, and G show SEM images of the upper epidermis of leaf sheaths indicative of epidermis cells and a few stomata of KPS01-12, KK3, UT84-12 and LK92-11, respectively. Figures 1 B, D, F, and H show SEM images of the lower epidermis of leaf sheaths of the four cultivars indicative of epidermal cells, stomata, trichomes, lots of cork cells and numerous silica cells. Figure 1 SEM images of the leaf sheath surface of 4 Thai sugarcane cultivars. A: Upper epidermis of KPS01-12leaf sheath, B: lower epidermis of KPS01-12 leaf sheath, C: Upper epidermis of KK3 leaf sheath of, D: lower epidermis of KK3 leaf sheath, E: Upper epidermis of UT84-12 leaf sheath, F: lower epidermis of UT84-12 leaf sheath, G: Upper epidermis of LK92-11 leaf sheath, H: lower epidermis of LK92-11 leaf sheath (bt=bicellular trichome, ep= epidermis cell, sc= silica cell, st=stomata, ut=unicellular trichome)

206 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry CO-PRODUCTS Figure 2 SEM images of the lower epidermis layer of the leaf sheath surface of KK3 cultivars showed cork cells (cc), epidermal cells (ep), silica cells (sc), stomata (st) and unicellular trichome (ut). The EDS analysis of the upper and lower epidermis layers of the leaf sheath surface shows that the lower epidermis layer contains more silicon content than the upper epidermis layer. Moreover, the silicon content was also different in each cultivar as shown in Table 1. The element maps of silicon on the leaf sheath surface of the 4 cultivars were in the same trend. On the upper epidermis layer had silicon content less than 1 wt% and on the lower epidermis layer had silicon content more than 6 wt%, as shown in Table 1. The structural characteristics of the lower epidermis layer included numerous silica cells which was a source of silica. While the upper epidermis layer had only a few silicon contents. The distribution of silicon on the lower epidermal layer of the leaf sheath of 4 cultivars are shown in the X-ray mapping of EDS. Silicon appeared on the silica cells, surrounding stomata and some epidermal cells. Figure 3 A shows EDS spectrum indicating that the lower epidermal layer of leaf sheath of KPS01-12 contained silicon (Si), oxygen (O), and carbon (C). Figure 3 B, E, H and K are SEM images of the lower epidermal layer of KPS01-12, KK3, UT84-12 and LK92-11, respectively. Figure 3 C, F, I and L are X-ray maps of Si of KPS01-12, KK3, UT84-12 and LK92-11, respectively. Figure 3 D, G, J, M are the x-ray map of O of the four cultivars, respectively. The results indicated that Si and O had similar distributions and were likely to form SiO2 in silica cells, around stomata, and epidermal cells. The high intensity of Si on the X-ray map showed in the silica cell position, on the other hand, silicon was not found in the trichome area (Figure 3 C). Paopun et al. (2022) reported that the leaf surface of the Suphanburi 50 cultivar of sugarcane had silicon around stomata, bulliform cells and in silica cells. Kumar et al. (2017) reported that in grasses, silica deposited in epidermal cells of inflorescence bracts, epidermal cells of leaf and root endodermis. Mvondo-She et al. (2019) found that in citrus, silica was accumulated in the leaf and increased with leaf age. Table 1 Silicon content (wt%) in the upper epidermis and lower epidermis in leaf sheath of 4 Thai sugarcane cultivars, analyzed by EDS combined with FE-SEM Note: The numbers followed by the same letters in the column (a, b, c) are not significantly different with a confidence level of 0.05

207 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry CO-PRODUCTS Figure 3 Analysis of the lower epidermis of sugarcane leaf sheath surface. A.) EDS spectrum of the lower epidermis of KPS01-12 sugarcane leaf sheath surface. B), E), H), K) SEM images of the lower epidermis of KPS01-12, KK3, UT84-12, LK92-11 sugarcane leaf sheath surface, respectively. C), F), I), L) Silicon X-ray maps of the above samples. D), G), J), M) Oxygen X-ray maps of the above sample, respectively. EDXRF analysis of powder from the dry leaf sheath of the 4 cultivars shows the differences of silicon content, summarized in Table 2. Silicon content in KPS01-12 was similar to KK3 (about 4%wt). Which was higher than silicon in UT84-12 and LK92-11.

208 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry CO-PRODUCTS Table 2 Silicon content (wt%) in the leaf sheath of the 4 cultivars, analyzed by EDXRF Note: The numbers followed by the same letters in the column (a, b, c) are not significantly different with a confidence level of 0.05 Note that the XRF result of the same KPS01-12 cultivar indicates an average value of Si content in a large volume of interaction while SEM/EDS provides a higher spatial resolution (a specific area of the sample) with images. It is because XRF using a relatively large X-ray incident beam to analyze a sample, compared to SEM/ EDS which used a much smaller electron incident beam. EDXRF analysis shows that the accumulated silicon in the leaf sheath of the four cultivars were close to the silicon content in the sugarcane leaf of the Suphanburi 50 cultivar, reported by Paopun et al. (2022) it was reported that the silicon content in the leaves were about 4.44-6.34 wt% in the apical part of a leaf, 4.07-5.39 wt% in the middle and 2.65-3.61 wt% in the base. The accumulation of silicon in sugarcane may involve other factors such as the water solubility of silica, pH of the soil [22], silicon content in the soil, solubility of silicon compounds in water, and the ability of silicon absorption capacity of each plant. Tubana et al. (2016) reported that most of monocotyledons, such as sugarcane, barley, rice and wheat, can accumulate large amount of silicon. Moreover, plants can accumulate silicon in the epidermis cell and phytoliths, especially grasses and sedges accumulated high concentrations of silicon in the phytoliths of leaves [24]. Fox et al. (1969) reported that silicon was deposited in the leaves and leaf sheaths more than in roots. The amount of silica content in the sugarcane leaf sheath was higher than in the leaf blade, which is in good agreement with this study. We demonstrated that the leaf sheaths of sugarcane, which are agricultural wastes could be used to extract bio-silica. The bio-silica can be beneficial in several applications such as composite materials, pharmaceutical industries [26], and blended Portland cement [27]. This approach can increase the value of agricultural waste for agriculturists and reduce air pollution from burning waste in the field. Sugarcane leaves are another part of sugarcane that are interesting silica deposits to study for silica content. 4. CONCLUSIONS Silicon accumulated in the form of silica in the leaf sheath of four Thai sugarcane cultivars was investigated. There were differences in silicon content in each cultivar. KK3 had the highest silicon content of 4.10±0.27 wt% when analyzed by EDXRF, which was close to silicon in KPS01-12. These were followed by UT84-12 and LK92-11, respectively. SEM&EDS showed that the content of silicon in the lower epidermal layer was higher than in the upper epidermal layer because there were silica cells on the lower epidermal layer and not on the upper surface of the leaf sheath. EDXRF technique used a large X-ray incident beam and analyzed relatively much larger volume of interaction. It is good to provide an average Si content of the leaf sheath. While SEM &EDS analysis used a small incident electron beam which provided a higher spatial resolution of the leaf sheath (specificity of the area of analysis). This study can be used to guide the process of extracting bio-silica from the 4 cultivars of sugarcane leaf sheath in order to add value of the agricultural waste. 5. ACKNOWLEDGEMENT The authors wish to thank the Kasetsart University Research and Development Institute, Bangkok, Thailand and COAX Group Corporation Ltd., Bangkok, Thailand for the equipment support. We are grateful to Prof. S. Seraphin for the fruitful discussion on manuscript preparation. 6. REFERENCES [1] Kew royal botanical garden, plants of the world online, 1753, http://powo.science.kew.org/taxon/urn:lsid:ipni. orgnames:419977-1(cited online 1 November 2022). [2] Athipanyakul T., Choonhawong K. and Potchanasin C. (2020). The challenge for Thai sugarcane farmers. Food and Fertilizer Technology Center for the Asian and Pacific Region, https://ap.fftc.org.tw/article/1840 (cited online 15 May 2023). [3] Motomura, H., Hikosaka, K. and Suzuki, M. (2008). Relationships between photosynthetic activity and silica accumulation with ages of leaf in Sasa veitchii (Poaceae, Bambuoideae). Annals of Botany, 101, pp. 463-468, DOI: 10.1093/aob/mcm301.

209 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry CO-PRODUCTS [4] Sornpoon, W., Bonnet, S., Kasemsap, P., Prasertsak, P. and Garivait, S. (2014). Estimation of emission from sugarcane field burning in Thailand using bottom-up country-specific activity data. Atmosphere, 5, pp. 669-685, DOI: 10.3390/atmos5030669. [5] Poltam, S., Kaewrueng, S., Duangpatra, P., Weerathaworn, P. and Sanglestsawai, S. (2018). Assessment of biomass loss and air pollution caused by pre-harvest sugarcane burning using the closed loop combustion system model. Environment Asia, 11(2), pp. 1-8, DOI: 10.14456/ea.2018.18. [6] Sahebi, M., Hanafi, M.M., Akma,r A.S.N., Rafii, M.Y, Azizi, P., Tengoua, F.F., Aswa, J.N.M. and Shabanimofrad, M. (2015). BioMed Research International, 2015, pp. 1-16, DOI: 10.1155/2015/396010. [7] Massey, F.P., Enos, A.R. and Hartley, S.E. (2006). Silica in grasses as a defense against insect herbivores: contrasting effects on folivores and a phloem feeder. Journal of Animal Ecology, 75, pp. 595-603. [8] Hunsigi, S. (1993). Production of Sugarcane Theory and Practice. New York: Springer-Verlag. [9] Exley, C., Guerriero, G. and Lopez, X. (2020). How is silica acid transported in plants?. Silicon, 12, pp. 2641-2645, DOI: 10.1007/s12633-019-00360-w. [10] Prasad, R. (2019). Silicon in plant structure and inorganic c-sequestration. International Journal of Plant Environment, 5(2), pp. 67-77. [11] Vandegeer, R., Zhao, C., Cibils-Stewart, X., Wuhrer, R., Hall, C.R., Hartley, S.E., Tissue, D.T. and Johnson, S.N. (2021). Silicon deposition on guard cells increases stomatal sensitivity as mediated by K+ efflux and consequently reduces stomatal conductance. Physiologia Plantarum, 171(3), pp. 358-370, DOI: 10.1111/ ppl.13202. [12] Matichenkov, V.V. and Calvert, D.V. (2002). Silicon as a beneficial element for sugarcane. Journal American Society of Sugarcane Technologists, 22, pp. 21-29. [13] Kaufman, P.B., Dayanandan, P., Franklin, C.I. and Takeoka, Y. (1985). Structural and function of silica bodies in the epidermal system of grass shoots. Annals of Botany, 55, pp. 487-507. [14] Paopun, Y., Thanomchat, P., Toyen, D., Changjan, D. and Sinbuatong, N. (2021). Silica on leaf surface of supanburi50 cultivar sugarcane of Thailand. In the 38th MST International Conference, pp. 86-87. March 23-26, 2021, Chonburi, Thailand. [15] Clements, H.F. (1980). Sugarcane Crop Logging and Crop Control Principle and Practices. The United States of America: The University Press of Hawaii. [16] Paopun, Y., Thanomchat, P. and Toyen, D. (2022). Analysis of biosilica in sugarcane leaves. Microscopy and Microanalysis Research, 35(2), pp. 5-9. [17] Alves, R.H., Reis, T.V.S., Rovani, S. and Funfaro, D.A. (2017). Green synthesis and characterization of biosilica produced from sugarcane waste ash. Journal of Chemistry, 2017, pp. 1-9, DOI: 10.1155/2017/6129035. [18] Goldstein, J.I., Romig Jr., A.D., Newbury, D.E., Lyman, C.E., Echlin, P., Fiori, C., Joy, D.C. and Lifshin, E. (1992). Scanning Electron Microscopy and X-Ray Microanalysis: A Text for Biologists, Materials Scientists, and Geologists. New York: Plenum Press. [19] Moore, P.H. (1987). Anatomy and morphology. In D.I. Heinz, (Ed), Developments in Crop Science Book Series Volume11. ScienceDirect online: Elsevier B.V., 85-142. [20] Kumar, S., Soukup, M. and Elbaum, R. (2017). Silicification in grasses: variation between different cell types. Frontiers in Plant Science, 8(438), pp. 1-8, DOI: 10.3389/fpls.2017.00438. [21] Mvondo-She, M.A. and Marais, D. (2019). The investigation of silica localization and accumulation in Citrus. Plants, 8(200), pp. 1-12, DOI: 10.3390/plants8070200. [22] Barber, D.A. and Shone, M.G.T. (1965). The absorption of silica from aqueous solutions by plants. Journal of Experimental Botany, 17(52), pp. 569-578. [23] Tubana, B.S., Babu, T. and Datnoff, L.E. (2016). A review of silica in soils and plants and its role in US agriculture: history and future perspectives. Soil Science, 181(9/10), pp. 393-411. [24] Klancnik, K., Vogel-Mikus, K. and Gaberscik, A. (2014). Silicified structures affect leaf optical properties in grasses and sedge. Journal of Photochemistry and Photobiology B: Biology, 130, pp. 1-10. [25] Fox, R.L., Silva, J.A., Plucknett, D.L. and Teranishi, D.Y. (1969). Plant and Soil, 30(1), pp. 81-92. [26] Singh, J., Boddula, R. and Jirimali, H.D. (2020). Utilization of secondary agricultural products for the preparation of value added silica materials and their important applications: a review. Journal of Sol-Gel Science and Technology, DOI: 10.1007/s10971-020-05353-5. [27] Guzman, A., Gutierrez, C., Amigo, V., Mejia de Gutierrez, R. and Delvasto, S. (2011). Pozzolanic evaluation of the sugar can leaf. Materiales de Construccion, 61(302), pp. 213-225, DOI: 10.3989/mc.2011.54809.

210 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry CO-PRODUCTS Lalita Pooddeea , Mallika Boonmee Kongkeitkajornb,c, Koji Kobayashid , Shigeyuki Funadad , Katsushige Yamadad , Prawphan Yuvadetkuna , Atsushi Minaminoa , Tatsuya Matsunoa * a Cellulosic Biomass Technology Co., Ltd., Kumphawapi, Udon Thani, Thailand, 41110 b Department of Biotechnology, Khon Kaen University, Khon Kaen, Thailand c Fermentation Research Center for Value Added Agricultural Products, Khon Kaen University, Khon Kaen, Thailand, d Toray Industries, Inc., New Frontier Research Laboratories, Kamakura, Japan *Correspondence to: [email protected] ABSTRACT: Agricultural residue has the potential to be used as substrate for many industrial processes. Sugarcane bagasse is a one of agricultural residue from the sugar industry. In 2022/2023, 93.9 million metric tons of sugarcane was produced in Thailand [1]. The sugarcane bagasse residue from sugar factory is mainly used as fuel in boiler to produce electricity releasing waste gas contains PM 2.5 in case of incomplete combustion. This poster shows alternative way to increase the sugarcane bagasse value rather than only use in boiler by transforming into high value-added product from cellulose, hemicellulose and lignin to cellulosic sugar, oligosaccharide, polyphenol, and solid polyphenol. Cellulosic Biomass Technology Co., Ltd. (CBT) produces high purified cellulosic sugar from sugarcane bagasse by novel multi-membrane integrated system. This demonstration project entailed Toray Industries, Inc (Toray) verifying a process to separate, purify, and concentrate cellulose-derived sugars in non-edible biomass. It leveraged a membrane-based bioprocess that combines the Toray’s water treatment membrane technology and enzymes that employ biotechnology. Toray undertook this effort at a demonstration facility in Thailand as part of a project that the New Energy and Industrial Technology Development Organization (NEDO) and National Innovation Agency (NIA) are supporting. The company proved that carbon dioxide emissions from this process are less than half those of conventional production setups that concentrate sugar solutions by evaporating water. Cellulosic sugar’s performance was evaluated by using it as substrate for ethanol and lactic acid production. In the case of ethanol fermentation test by Saccharomyces cerevisiae, ethanol concentration increased when increasing glucose concentration of cellulosic sugar up to 200 g/L with the maximum concentration as 77 g/L(average)with the ethanol yield 0.43 g/g.Forlactic acid fermentation byLactobacillus delbreuckii, themaximumlactic acid produced is 67 g/Lwhen using initial glucose concentration of cellulosic sugar at 100 g/L (average) but the maximum yield is 0.77 g/g when using initial glucose concentration at 50 g/L (average). So, based on the result, cellulosic sugar has the potential to be used as substrate for ethanol and lactic acid production, besides, the production efficiency depends on microorganism strain. Moreover, cellulosic sugar has a possibility to be applied for another biofuel and biochemical production. Keyword: Cellulosic sugar, Sugarcane bagasse, Ethanol, Lactic acid, Membrane P-036 Evaluation of cellulosic sugar from sugarcane bagasse produced by novel multi-membrane integrated system for biofuel and biochemical production Reference [1] Office of The Cane and Sugar Board (2023). Cane Area and Yield 2022-2023. Retrieved 10 June 2023 from https://www.ocsb.go.th. gyp

211 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry CO-PRODUCTS Buchita Penjana , Massalin Nakphaichitb , Koji Kobayashic , Shigeyuki Funadac , Katsushige Yamadac , Metakarn Leartkiatratchataa , Atsushi Minaminoa , Tatsuya Matsunoa,* a Cellulosic Biomass Technology Co., Ltd., Udon Thani, Thailand b Department of Biotechnology, Faculty of Agro-Industry, Kasetsart University, Bangkok, Thailand. c Toray Industries, Inc., New Frontier Research Laboratories, Kamakura, Japan *Correspondence to: [email protected] ABSTRACT: Agricultural residues like sugarcane bagasse is low-value byproducts of sugar industry. Cellulosic Biomass Technology Co., Ltd. developed the technology for sugarcane bagasse as a raw material to produce value added products. Solid polyphenol is one of the products from sugarcane bagasse produced through pretreatment and solid-liquid separation. Solid Polyphenol contains diatomaceous earth and phenolic compounds with demonstrated antioxidant properties for supplement of livestock feed. In our study, we investigated the effect of solid polyphenol supplementation on the performance of swine and poultry. Solid Polyphenol showed the capability of post weaning piglets at 100 - 400 ppm added by increasing average daily gain (ADG) and improving feed conversion ratio (FCR) as well as diarrhea problem. As for poultry nutrition, Broiler chicken, analysis of data showed effects on Villi width in Gastrointestinal (GI). Moreover, supplemented solid polyphenol at 50 - 200 ppm in feed significantly improved ADG and FCR as well as carcass characteristic of broiler chicken by reducing abdominal fat contents. On Layer hens trial, supplemented solid polyphenol improve growth performance of laying hens under environmental stress condition by improving egg production and FCR at 50 - 200 ppm of solid polyphenol supplemented. These results indicated that Solid polyphenol can be considered as beneficial dietary supplement for improving growth parameters, and oxidative status in poultry and swine. P-037 Evaluation of Solid Polyphenol as a Livestock Feed

ECONOMIC AND MANAGEMENT 212 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry

213 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT Nuttapon Photchanaprasert1 * , Atchara Patoomnakul1 , Ravissa Suchato1 and Borworn Tanrattanaphong1 1 Department of Agricultural and Resource Economics Kasetsart University 50 Ngam Wong Wan Rd, Lat Yao Chatuchak Bangkok 10900 *Correspondence to: Department of Agricultural and Resource Economics Kasetsart University 50 Ngam Wong Wan Rd, Lat Yao Chatuchak Bangkok 10900, [email protected] ABSTRACT: Thailand ranks second in sugar exports after Brazil. Over the past two decades, sugar and by-product exports have increased annually by 7.66%. In 1998, raw sugar comprised 90.45% of sugar and by-product exports. Prior to 2002, Thai sugar exports contained less raw sugar. This has made refined white sugar and unprocessed sugar significant exports for Thailand. From 2004 to 2014, Thailand's sugar and product exports grew by 12.93% annually. Raw sugar had the highest annual growth rate at 19.29%, followed by white refined sugar, sugar confectionery, and bagasse. Thailand's main exports, raw sugar and white refined sugar, decreased after global agricultural commodity value chains and food shifted. Although exports of bagasse, other sugars, and sugar confections have increased, total exports of sugar and sugar products have decreased. The analysis of Thailand's sugar industry's global value chain competitiveness and upgrading strategy is important for sustainable development in the future. This study will employ Gereffi and Fernandez-Stark's (2016) Global Value Chain Framework, which addresses six critical issues: 1) the structure and performance factors of the value chain; 2) the geographic scope of the value chain; 3) the internal governance of the value chain; 4) upgrading strategies within the value chain; 5) the organizations and institutions within the country; and 6) the winners and losers in the domestic industry. Issues 1–3 will be examined in the context of global links, while issues 4–6 will be examined in the context of the country's activities. O-009 Thailand's sugar industry's global value chain competitiveness and upgrading strategy. 1. The potential of Thailand's sugar industry in the global value chain. 1) Competitiveness of Thailand's sugar industry Based on the percentage of raw sugar, refined white sugar, and molasses exported, it can be determined that Thailand's sugar industry is unable to meet the demand for sugar on the global market. This is in contrast to Brazil and India, whose industries can supply 65% and 11% of the demand, respectively. This demonstrates that, in comparison to its rivals, Thailand has a comparatively low capacity to supply the demand for raw sugar on the international market. Thailand's ability to satisfy market demand is lower than its rivals' in the case of the refined white sugar, as it can only supply 7.3% of the demand on the world market, compared to Brazil's 10.2% and India's 19.4%. Thailand's ability to satisfy the market need for the molasses industry is quite low when compared to India, as seen by the fact that it can only meet 1.9% of the worldwide market demand, while India can meet 28.9% of it and Brazil cannot. In conclusion, Thailand's ability to meet the demand of the global market is low in all three sectors of the country's sugarcane-sugar business due to the land area restriction that impacts its production capacity, making Thailand less competitive than its rivals. 2) The competitive situation in the global market. The Herfindahl-Hirschman Index (HHI) measures global competition in the sugar industry. Raw sugar, white sugar, and molasses sugar have HHI values of 0.32, 0.07, and 0.08, respectively. This suggests that Brazil, India, and Thailand dominate the worldwide raw sugar sector, which is highly concentrated or less competitive. Germany exports beet-based white sugar, and the UnitedArab Emirates'Al Khaleej Sugar and The Gulf Sugar produce 6,000 tons of white sugar per day, accounting for 3% of the world's production. Exporting refined white sugar from imported raw sugar. Exporters from India, Mozambique, Indonesia, Russia, Germany, Egypt, and others compete in

214 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT the low-concentration molasses sugar sector. In Thailand's main markets, Indonesia is the world's largest importer of raw sugar, and Thailand is the largest exporter, with an HHI of 0.26, indicating a low exporter concentration. India has the biggest export market share at 33.6%, while Thailand has 19.8%. Thailand's export prices are higher than those of competitors like Brazil, which has lower manufacturing costs and sells sugar at a lower price. Thailand's market share has declined. Italy, Sudan, and the US are the top importers of pure white sugar. The exporting countries' disruption levels in Italy and the US are 0.3, 0.66, and 0.18, indicating low competition in Italy and Sudan and moderate competition in the US. Germany exports the most to Italy, Mexico to the US, and India to Sudan at 80.4%. Thailand's limited part of the world pure white sugar market is due to Italy’s import of beet sugar, cheaper than sugarcane sugar, and Thailand's focus on selling sugar to Southeast Asia. The United States imports the most raw sugar, mostly from Guatemala. Thailand is the biggest net importer of refined sugar, mostly from India, due to a 75% drop in export growth in 2020–2021. Due to higher oil prices, ethanol production from imported raw sugar has increased. Imported raw sugar costs $185 per ton in the US. The UK imports raw sugar from India at $225 per ton, making it the third-largest importer. Thailand has no chance of competing in the global sugar market. Thailand has the potential to export 1.2 billion US dollars in raw sugar to Indonesia, the world's largest raw sugar import market, but only 878 million US dollars were exported in 2021. Thus, Thailand can export 294 million US dollars to Indonesia, where it has the greatest comparative advantage over Brazil and India. Brazil has the most export potential in China, with 973 million US dollars, but only 656 million were sold in 2021, followed by Cuba and Thailand. Brazil, Guatemala, and Thailand have the most export potential to the US. However, the Thai white sugar industry has the potential to export to Southeast Asian countries like Cambodia, Myanmar, and Vietnam due to geographical limitations and competition in the white sugar industry, which is made from beet sugar at a lower price than cane sugar, and because Thailand cannot export molasses due to ethanol demand. 3) Entry Barriers Indonesia charges an 8.66% MFN tax on raw sugar exports. Thailand's actual tax rate is 5%, compared to 8.66% for Brazil, India, and Australia. India exports cheaper than Thailand due to the government's backing of 6,000 rupees per ton, or 80 US dollars. Thailand pays 15% inside the quota (WTO, 19.45 million tons per year) and 50% beyond the quota in China, the same as its competitors. Thailand's 18.8% US tax rate is lower than Brazil's 56.39%. White sugar with a net weight that Thailand principally exports inside the ASEAN Trade in Goods Agreement (ATIGA) is tax-free, except for Indonesia (10%), the Philippines (5%), and Vietnam (5%). Thai pure raw sugar is taxed at 44.23% and supported at 4.65% in the Vietnamese market, whereas raw sugar is taxed at 29.23% and supported at 4.65%. Importing sugar cane bagasse costs competitors 85%. Japan and Indonesia both charge 8.38% for Thai sugar imports, while Taiwan charges 9.38%. Vietnam has a 5.1% tax rate due to its tax advantage over China. Raw sugar exported to Indonesia must meet 15 non-tariff conditions, including SPS and traceability. The US market must pass 41 measures, and China must pass 70. Indonesian raw sugar has the fewest non-customs duty measures. Cambodia requires four measures, Vietnam 46, and the Philippines 60 for refined white sugar exports. Cambodia has the fewest non-customs duties. Molasses must pass 15 measures for Japanese export, 15 for Malaysia, and 42 for Korea. The Most Favored Nation requires these standards for all nations. 2. Competitive factors 1) Characteristics of the production chain 1.1 Position in the production chain Thai sugarcane and sugar production are still at the bottom of the smiling curve. The country produces raw brown sugar from sugarcane. Some of it is turned into white sugar or ethanol (10% of sugarcane is utilized to make ethanol), but there is minimal research and development into new strains, growing methods, or higher-value goods. A low-value-added industry results. The biotechnology industry, which processes 1% of sugarcane yield, does not process into higher-value products. 1.2 Global value chain participation Thailand exports less raw sugar than Brazil. Thailand's second-ranked raw sugar business has deteriorated due to India's strong export expansion, supported by subsidies and stock clearance, resulting in lower production costs and prices. India's role in the global raw sugar chain is growing, while Thailand's participation in the valuable white sugar chain is low due to competition from European countries that produce white sugar from sugar beets and the world's largest sugar processing companies in Arab Emirates that import raw sugar for processing and export. Thai ethanol is made from molasses, a byproduct of raw sugar. 1.3 Length of value chain The value chain is growing.Technological advances have led to the production of bioproducts and high-value

215 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT processed sugar and molasses, making the modern sugarcane and sugar value chain longer. Thailand's processing is limited due to its early growth. Traders sell raw and white sugar to end users who use it in food and beverages. Due to few middlemen and minimal earnings, Thai sugarcane and sugar have a short value chain. Value chain management is cheap and information flows well. Thai sugar producers are efficient and can meet demand and buyer demands. 1.4 Governance The sugarcane and sugar sector has market governance. Due to the mild transaction complexity, Thai sugar exporters can simply communicate with sugar importers and follow sugar's set criteria. Due to unpredictable production lots, there is some complication. New production rules that address labor and environmental issues make transactions difficult. Data processing according to each new standard involvessignificant expenditure, but consumers may place specific sugar production orders that can be easily met. Sugar trading follows worldwide norms, making it easy to share production processes. Product development and improvement are entrepreneurial skills, such as turning sugar into syrup for the food business, low-calorie sugar for health-conscious consumers, or biogas for factory energy savings. Export sugar must meet client requirements for quantity, delivery time, quality, and reactivity. 2) Technology and R&D 2.1 Innovation and technology Brazil uses automation for cultivation and harvesting, while Thailand still uses manual labor. Thailand produces raw sugar and white granulated sugar at a basic level, similar to its competitors. However, ethanol production from sugarcane bagasse and sugar transformation into chemicals, bioproducts, cosmetics, and medicine is possible. Thailand's technology industry is still young due to a lack of supportive markets and knowledge held by huge international research and development businesses like Corbion Purac Cargrill, NatureWorks, and Total Corbion NatureWorks. Thus, Thailand falls behind in the high-end downstream sugar industry. 2.2 Research and development Business-level research and development in the sugar industry involves improving production processes to reduce factory costs, such as converting molasses from sugar production into bioenergy to reduce factory electricity consumption, and developing new products to meet modern consumer needs, such as converting sugar into syrup for the food industry or low-calorie sugar for health-conscious consumers. Thailand's Sugarcane Research Center, Office of Sugarcane and Sugar Board, and Ministry of Industry develop sugarcane cultivars upstream. However, it has not been very successful, and there are still efforts to push the sugar industry to a new S-curve, such as cooperation to create a bioeconomy under the "Sustainable Economy" project, future industrial cluster development, and prototype ethanol plants from sugarcane bagasse, but production costs are still high. Unlike Europe, India, and Brazil, Thailand's research and development is scattered among many bodies, making budget and efficiency unclear. 3) Upgrading • Economic Upgrading Product and process upgrades will boostThailand'ssugarsector. Due to a lack ofmarket demand and research and development, product upgrading mostly involves changing packaging and product design to meet customer demands. Process upgrading involves improving the production process by replacing humans with machines in some stages, such as using automatic packaging machines, robotic product sorting, and using sugar cane by-products to generate biomass electricity. • Social Upgrading Major sugar importers are now considering labor and environmental issues when buying sugar. Coca-Cola, PepsiCo, Unilever, and Hershey exclusively buy sugar with Bonsucro criteria, which compels sugar producers to improve their wage rates, labor benefits, labor rights, and greenhouse gas emissions. Thai entrepreneurs can improve society. 4) Business Environment 4.1Available raw materials According to the table of production factors in the country, the sugarcane and sugar industry relied on domestic raw materials at 85.04% and imported raw materials at 14.96%, indicating that Thailand has ready raw materials for the industry due to its favorable geographical location. Thailand has doubled its sugarcane cultivation and production since 1990, although most of the land is owned by small farmers, resulting in higher average costs than other countries. 4.2 Logistics and Transportation Since most of the growing area is small farms that cannot use sugarcane harvesters or large tractors, most of

216 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT the labor force is needed to harvest and transport sugarcane to sugar factories in Thailand. Sugarcane harvesting and transportation, which vary by region, require a logistics management system. This causes low harvesting and logistics efficiency, which lowers sugar sweetness and raises sugarcane harvesting and transportation expenses in Thailand, which account for 40% of sugarcane production costs. Brazil has greater sugarcane crops and lower harvesting and shipping costs than Thailand. 4.3 Government policies Since 1984, the Sugarcane and Sugar Act has regulated the industry to promote factory and consumer fairness and economic stability. Sugarcane and sugar distribution are tightly regulated. In 2018, Thailand amended the law to change the sugarcane and sugar industry by lifting the domestic sugar price ceiling, abolishing the sugar quota system, and suspending contributions to the fund. However, the draft of the Sugarcane and Sugar Act amendment, which covers other products like sugarcane juice and bagasse, has failed and caused conflicts between sugarcane farmers and factories, potentially affecting Thailand's participation in the value chain. 4.4 Demand condition Historically, 30% (2.6 million tons) of sugar is consumed locally and 70% is exported. Due to industrial and food and beverage industry investment, domestic consumption is rising. Sugar for home usage is the same to exporting sugar. Thai raw sugar is mostly imported. Bioplastics, biopharmaceuticals, and nutraceuticals are still young sectors in Thailand. Producers are not motivated to invest in research and development due to low demand and expensive prices. The sugarcane and sugar sector suffers from the government's failure to motivate producers and consumers. 3. Thailand's sugarcane industry development guidelines. To minimize production costs and compete in the global value chain, sugarcane cultivars with high yields, disease resistance, and local climatic compatibility must be developed. Grouping farmers for harvesting machinery saves logistics costs. Factory sugarcane water systems increase quality and cut wait times. The government should create a central sugarcane research and development body, market the bio-industry to customers, and aid private sector research. Keywords: Sugarcane and sugar industry, Global value chain, participation , upgrading

217 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT A.K. Sah1 and R. Viswanathan2 ICAR-Indian Institute of Sugarcane Research, Lucknow, India *Correspondence to: [email protected], [email protected] ABSTRACT: The Indian sugar industry was hugely impacted and about 70000 crore rupees was stucked in unsold sugar and ethanol during nationwide lockdown during the months of April- June, 2020 as an effort to contain the spread of COVID-19. And this has created huge liquidity crisis in the sector. The country witnessed the return journey of migrant labours bare-footed to their respective native villages. Some of the reports says that about 150 lakhs of migrants returned during lockdown and out of this more than 36 lakhs migrants returned to Uttar Pradesh and about 25 lakh returned to Bihar. All these migrants returned to villages where farming is pre-dominant economic activity, the farm sector can play bigger role in providing jobs and livelihood to the migrants. Like wise the potential available in sugarcane sector may be leveraged for livelihood of migrants. The Uttar Pradesh is largest state in terms of sugracne cultivation, and has the good potential to absorb major chunk of in-migrants for their livelihoods. During COVID lockdown 36 lakh migrants returned to the Uttar Pradesh, out of this 20.37 lakh (54%) were skilled workers and 3.19 lakh (about 9%) were women. One-third of these returnee women may be engaged in production and marketing of sugarcane nursery with bud chip or single cane node method, through formation of 5 Self Help Groups (SHGs) of 10-20 women in each SHG at each of 125 selected Cane Development Council in the state. About50% of cane raea in the UP state (roughly 13 lakh ha) is being planted in september-march every year. If 10% of the planting to be done by nursery raised plants (90% with traditional method) requires 3900 million plants and this may create employment for about 3.25 lakh women for 30 days. By selling these plants @ Rs.3.0 each, they can collectively earn profit to tune of Rs.325 crore, thus giving opportunity to each women to earn Rs.10000 in just 30 days time. More than 20 lakh returnee in the UP state are skilled workers, they may be employed in creating value chain in diversified traditional food products like gur, khandsary and more over production of nutritionaly fortified gur, chikki, candy etc. in different shape, size, packaging. If we take into account the country as whole there are about 1.0 lakh jaggery units. Considering direct employement of 20 labour in each of the unit, 20 lakh jobs per day potential is there in this sector for at least six months (November to April) in a year. Jaggery manufacturer generally give wages @ Rs300-350 per day. This way each one of them earn Rs.9000-10500 per month and Rs.54000 to Rs.63000 in six months. And this sector in the state has the potential to absorb about 30-40 percent of 20.37 lakh skilled migrants returned during this pandemic. The employment for about 5.0 to 6.0 lakh un-skilled person can be created under intercropping system. In Uttar Pradesh, the green cane top availability lies between 136-182 lakh tonne, good proportion of green top is being used as green fodder for milch animals, and rest is spoiled or wastage. Returnee migrants may be engaged locally in Pitt digging for making silage from left over green cane top. Returnee migrants may be trained in making packaging material, serving plates, spoon, cup from sugarcane baggase. The investment in this sector under the MSMEs may be explored and mobilized. The Value-Chain for product diversification in sugarcane should be established in rural settings. O-015 Livelihood opportunity for migrants in sugarcane sector 1 Principal Scientist, 2 Director

218 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT Panatda Utaranakorn1,2 and Patcharee Suriya1,2* 1 Department of Agricultural Economics, Faculty of Agriculture, Khon Kaen University, Khon Kaen, 40002, Thailand 2 Northeast Thailand Cane and Sugar Research Center, Khon Kaen University, Khon Kaen, Thailand *Correspondence to: Department of Agricultural Economics, Faculty of Agriculture, Khon Kaen University, 123 Moo 16 Mittraphap Rd. Muang District, Khon Kaen, 40002, Thailand. E-mail: [email protected] O-034 An economic analysis of two baling systems for sugarcane straw collection: A case study in Khon Kaen province, Thailand 1. INTRODUCTION Khon Kaen province has the second highest production of sugarcane in the Northeast of Thailand (Office of the Cane and Sugar Board, 2022), and is a pilot province for implementing the BCG model of sugarcane production. The practice of green sugarcane harvesting and the reduction of burning sugarcane straw has been actively promoted in this area. Over the past few years, there has been a substantial surge in the demand for sugarcane straw used in biomass power generation in Khon Kaen. The demand has experienced a remarkable growth, rising from 8,000 tons in the 2019/20 production year to 51,500 tons in the 2021/22 production year. Consequently, local sugarcane farmers have invested in machines to collect sugarcane straw and supply it to biomass power plants for sale. However, these investments require a substantial amount of capital to acquire various machinery and equipment, including tractors, sweeping machines, balers, loaders, and trucks. Prior to making the investment decision, it is essential to conduct an economic analysis. This study aimed to analyze the costs and revenues, as well as assess the investment feasibility, of two baling systems used for collecting sugarcane straw. 2. MATERIALS AND METHODS This research relied on primary data on the production year 2021/22 obtained from in-depth interviews with 10 out of 12 farmers who owned a sugarcane straw baling machine in Khon Kaen province. The sample group represented 83.33% of the population. The survey was conducted through face-to-face interviews using a structured questionnaire. The data on sugarcane straw collection were analyzed using cost-benefit calculation, and financial feasibility assessment, considering six indicators: Payback Period (PB), Net Present Value (NPV), Benefit-Cost Ratio (BCR), Internal Rate of Return (IRR), Switching Value Test of Costs (SVTC), and Switching Value Test of Benefits (SVTB). 3. RESULTS AND DISCUSSION In Khon Kaen, two types of baling systems for collecting sugarcane straw have been found: the large square bale system and the round bale system. The process of collecting sugarcane straw involved several procedures, including sweeping, baling, loading, and transporting. To facilitate this process, farmers were required to make substantial investments in various machinery and equipment, such as tractors, sweeping machines, balers, loaders, and trucks. The findings showed that the total investment for sugarcane straw collection using the large square bale system amounted to approximately 203,326.51 USD, whereas the investment for the round bale system totaled around 134,434.90 USD. The analysis of costs, revenues, and investment feasibility for sugarcane straw collection was conducted based on the types of baling systems, and the results are as follows: 3.1 Costs and revenues of sugarcane straw collection The costs of sugarcane straw collection varied based on the types of baling systems. In the study area, it was found that the large square bale system had a lower cost per ton compared to the round bale system. Among the total costs, fuel and labor costs played a significant role and varied across different procedures of the collection process, including sweeping, baling, loading, and transporting as shown in Table 1. In the case of the large square bale system, the total fuel and labor costs amounted to 10.02 USD/ton. The baling procedure incurred the highest fuel and labor costs at 4.77 USD/ton, followed by transportation at 2.69 USD/

219 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT ton, loading at 1.38 USD/ton, and sweeping at 1.19 USD/ton. This finding was consistent with a study conducted in Brazil on the cost of sugarcane straw recovery, which indicated that the cost of straw baling accounted for the highest proportion (Sampaio et al., 2019). Similarly, the round bale system had a total fuel and labor cost of 12.26 USD/ton. The baling procedure incurred the highest fuel and labor costs at 5.12 USD/ton, followed by transportation at 3.66 USD/ton, loading at 2.24 USD/ ton, and sweeping at 1.24 USD/ton. Table 1. Labor and fuel costs of sugarcane straw collection classiflied by different procedures of the collection process in the production year 2021/22 in Khon Kaen, Thailand. The total costs and revenues for sugarcane straw collection showed in Table 2. The large square bale system incurred a total cost of 19.28 USD/ton, which comprised a variable cost of 12.65 USD/ton and a fixed cost of 6.63 USD/ton. The sugarcane straw bale was priced at 28.52 USD/ton, resulting in a net profit of 9.23 USD/ton. Annually, farmers collected approximately 2,243.33 tons of sugarcane straw bales, generating an annual revenue of 63,975.88 USD. The annual cost totaled 43,260.91 USD, leading to an annual profit of 20,714.97 USD. In the case of the round bale system, the total cost of collecting sugarcane straw amounted to 22.78 USD/ton, which comprised a variable cost of 14.16 USD/ton and a fixed cost of 8.62 USD/ton. The sugarcane straw bale was priced at 28.52 USD/ton, resulting in a net profit of 5.74 USD/ton. Annually, farmers collected approximately 1,000 tons of sugarcane straw bales, generating an annual revenue of 28,518.22 USD. The annual cost totaled 22,780.31 USD, leading to an annual profit of 5,737.91 USD. Despite the higher total annual cost of sugarcane straw collection using the large square bale system compared to the round bale system, the net profit achieved by the large square bale system was higher. These findings were consistent with a study conducted in Sa Kaeo province by Wongwaiwiriya et al. (2018), which demonstrated that the large square bale system yielded higher net profits due to its faster operation and ability to handle a larger quantity of sugarcane baling. Similarly, Zekic et al. (2010) found that the cost of baling biomass, square bale showed up to 28% cheaper than round bale. On the other hand, a study conducted in Brazil revealed that the total cost of round bale production was lower compared to square bale production. This cost advantage was primarily attributed to a high harvested straw productivity in the round bale system (Lemos et al., 2014). In addition, Sampaio et al. (2019) stated that the cost of sugarcane straw collection with the bale system depended on the amount of straw per hectare. When there was a low amount of straw per hectare, it resulted in lower operational efficiency of machinery, leading to increased the straw collection cost.

220 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT Table 2. Costs and revenues of sugarcane straw collection in the production year 2021/22 in Khon Kaen, Thailand. 3.2 Investment feasibility analysis for sugarcane straw collection The cash flows of costs and revenues ofsugarcane straw collection for 10 years were estimated under assumptions that variable costs and revenues increased by 10% per year. The present value of costs and revenues were adjusted by the discount rate of 6.5%, which was the minimum retail rate (MRR) charged for high-class retail customers by the Bank for Agriculture and Agricultural Cooperatives (2022). The result indicated that investment for both types of baling systems in Khon Kaen, Thailand were acceptable and worthy. The investment of the large square baling system provided the best performance with a payback period of 5.41 years, a net present value of 202,404.25 USD, a benefit-cost ratio of 1.36, and an internal rate of return on investment of 19.45% (Table 3). Additionally, the switching value test of costs and benefits were performed as well. The cost can increase as high as 36.07%, and the revenue can reduce to up to 26.51%. While the round baling system had a payback period of 7.78 years. A net present value of the investment was 43,674.87 USD, a benefit-cost ratio was 1.14, and an internal rate of return on investment was 11.06%. The cost can increase as high as 14.08%, and the revenue can reduce to up to 12.34%. The findings of this study were in line with the research conducted by Wongwaiwiriya et al. (2018) on the feasibility of investing in sugarcane leaf collection in Sa Kaeo province. Their study indicated that the round bale, small square bale, and large square bale systems all showed positive net present values (NPVs). However, it is important for farmers to consider their investment budget and the annual amount of sugarcane straw collection when deciding on the most suitable system for their needs. Table 3. Feasibility investment of sugarcane straw collection in Khon Kaen, Thailand. 4. CONCLUSION The cost of collecting sugarcane straw varied by the bale shape. The round bale had a total cost of 22.78 USD per ton, including a variable cost of 14.16 USD per ton and a fixed cost of 8.62 USD per ton. The net profit per ton was 5.74 USD, based on a selling price of 28.52 USD per ton. The large square bale had a total cost of 19.28 USD per

221 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT ton, including a variable cost of 12.65 USD per ton and a fixed cost of 6.63 USD per ton. The net profit per ton was 9.23 USD, based on a selling price of 28.52 USD per ton. With a 6.5% discount rate, the feasibility study revealed that investing in the large square baling system for sugarcane straw collection was more worthwhile than the round baling system. The large square baling system had a net present value of 202,404.25 USD, a benefit cost ratio of 1.36, an internal rate of return of 19.45%, and a payback period of 5.41 years. On the other hand, the round baling system had a net present value of 43,674.87 USD, a benefit cost ratio of 1.14, an internal rate of return of 11.06%, and a payback period of 7.78 years. In order to choose the most appropriate baling system, farmers need to consider factors such as their investment budget and the annual quantity of sugarcane straw to be collected. 5. ACKNOWLEDGMENT This research was funded by the National Research Council of Thailand (2021). The authors would also like to thanks to the Northeast Thailand Cane and Sugar Research Center (NECS), Faculty of Agriculture, Khon Kaen University, Khon Kaen, Thailand, for their assistance in conducting the field survey. 6. REFERENCES Bank for Agriculture and Agricultural Cooperatives. Minimum Retail Rate (MRR) for farmers in 2022 [Internet]. 2022 [cited 2022 Dec 20]. Available from: https://www.baac.or.th/file-upload-manual/rate/2565/Loan_ rate13092565.pdf. Lemos. S.V., M.S. Denadai, S.P.S. Guerra, M.S.T. Esperancini, O.C. Bueno, and I.C. Takitane. 2014. Economic efficiency of two baling systems for sugarcane straw. Industrial Crops and Products., 55: 97-101. Office of the cane and sugar board. Report of sugarcane planting area, production year 2021/22 [Internet]. 2022 [cited 2022 May 15]. Available from: https://w2.ocsb.go.th/wp-content/uploads/2023/04/. Sampaio. I.L.M., T.F. Cardoso, N.R.D. Souza, M.D.B. Watanabe, D.J. Carvalho, A. Bonomi, and T.L. Junqueira. 2019. Electricity production from sugarcane straw recovered through bale system: assessment of retrofit projects. BioEnergy Research., 12: 865–877. Wongwaiwiriya. J., K. Kuldilok, and T. Athipanyakul. 2018. Investment analysis of sugarcane leaves collecting business for biomass fuel case study: Sa Kaeo province. Available: http://mab.eco.ku.ac.th/wp-content/ uploads/2018/11/jiraporn-wongwiwiriya.pdf. Accessed Oct. 15, 2021. Zekic, V., V. Rodic, M. Jovanovic. 2010. Potentials and economic viability of smallgrain residue use as a source of energy in Serbia. Biomass Bioenergy., 34(12): 1789–1795.

222 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT Patcharee Suriya1,2 and Panatda Utaranakorn1,2* 1 Department of Agricultural Economics, Faculty of Agriculture, Khon Kaen University, Khon Kaen, 40002, Thailand 2 Northeast Thailand Cane and Sugar Research Center, Khon Kaen University, Khon Kaen, Thailand *Correspondence to: Department of Agricultural Economics, Faculty of Agriculture, Khon Kaen University, 123 Moo 16 Mittraphap Rd. Muang District, Khon Kaen, 40002, Thailand. E-mail: [email protected] O-035 Sugarcane leaf management and utilizations of farmers in Khon Kaen Province, Thailand 1. INTRODUCTION Sugarcane leaves provide significant benefits in both agriculture and industry. For instance, maintaining sugarcane leaves on the fields as blanket can improve soil structural quality through preservation of soil moisture and potential increasing of soil carbon and nitrogen (Ferreira-Leitão et al., 2010; Marin et al., 2014; Menandro et al., 2017; Castioni et al., 2018). The mulching leaves/straws can provide high soil moisture (Awe et al., 2015). In term of industry, Alves et al. (2015) proposed that keeping about 10 percent and 50 percent of leaves/straws in the field can increase the surplus electricity power above 23 percent and 102 percent. Sampaio et al. (2019) stated that utilization of leaves/straws as a complementary for biomass generation could increase the surplus electricity by 22 percent when the sugarcane leaves/straws are recovering about 9 percent in the field. In addition, the leaves/straws can be utilized as a source of bioenergy generation, including solid biofuel, bioethanol, and bio-methane (Ferreira-Leitão et al., 2010; Go and Conag, 2019). The sugarcane leaves serve as valuable materials for various purposes in several countries, including Thailand. For example, Yoddamnern and Yotwinyuwong (2020) mentioned that charcoal briquette from sugarcane leaves and bagasse has the highest thermal efficiency up to about 46.50 percent; and it can offer an income of 2,455.25 USD per year (1 USD = 35.1899 baht). Jankaew and Ariyakuare (2021) also researched in the development of sustainable utilization processes of sugarcane leaves for the design of products to be eco-efficient. Although, the literature on sugarcane leaf utilization is available worldwide, a comprehensive understanding of the available quantity of sugarcane leaves and the farmers’ management practices in Thailand is limited. In contrast, the sugarcane leaves/ straws are still burnt in the field after harvesting, which causes fly ash, silicate-dominated particle matter, and raises environmental concerns (Chandel et al., 2012; Kaewpradit, 2021). To ensure the sustainability of sugarcane leaf utilizations, this research aims to carry out an estimating of the supply and demand of sugarcane leaves in Khon Kaen Province of Thailand and a study of the sugarcane leaf management and its utilizations by farmers in Khon Kean which is one of the largest sugarcane productions in the Northeastern Thailand, which produces about 13.27 percent (6,130,803 tons) of total sugarcane production in the Northeastern Thailand in the production year of 2021/22 (Office of Agricultural Economics, 2022). Moreover, three biomass power generations are located in Khon Kaen, of which requires the sugarcane leaves for biomass power generation initially. 2. MATERIALS AND METHODS Thisresearch was based on primary and secondary data with regard to the sugarcane production year of 2021/22. The data ofsupply and demand ofsugarcane leavesin Khon Kaen was gathered from the interviews of three industries of biomass power generation, including both closed- and opened-ended questions. Additionally, the data related to famers’ management practices and utilization of the sugarcane leaf were collected through face-to-face interviews with 70 farmers using a structured questionnaire. Purposive sampling method was applied to select the participants, who sold sugarcane and sugarcane leaves during the sugarcane production year of 2021/22. In all, 70 farmers had planted sugarcane in Khon Kaen in the Northeastern region of Thailand. Descriptive statisticssuch asmean,frequency and percentage were utilized to describe the farmers’characteristics as well as various operations performed in the sugarcane leaf management and its utilization.

223 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT 3. RESULTS AND DISCUSSION 3.1 Supply and demand of sugarcane leaves in the sugarcane production year of 2021/22 In production year of 2021/22, about 3,337,795.84 tons of fresh sugarcane were produced which was 63.10 percent of total sugarcane production in Khon Kaen (Table 1.). The estimated supply of sugarcane leaves for both agricultural and industrial purposes was 333,779.58 tons. In contrast, the total demand for the sugarcane leaves was only 51,500 tons with about 15.34 percent of the total supply sugarcane leaves in the city. As most of biomass power industries did not use the sugarcane leaves as a complementary of biomass power generation, this suggests that approximately 282,279.58 tons of sugarcane leaves were not utilized for commercial. Table 1. Quantity of fresh sugarcane, burnt sugarcane, total sugarcane production, and estimated sugarcane leaves in Khon Kaen, in the sugarcane production year of 2019/20 - 2021/22. Source: 1) Office of The Cane and Sugar Board (2021) The demand of sugarcane leaves had increased more than twofold in the production year of 2021/22, comparing to the year of 2020/21 (Table 2.). This indicates that the sugarcane leaves were accepted as valuable raw material from the biomass power industries. The commercial utilization was also enlarged. The prices of sugarcane leaves from supplier start from 19.89 USD per ton to 28.42 USD per ton. These prices encourage some sugarcane producer to supply sugarcane leaves to the industries without burning leaves. However, most of them still use for other utilizations, including burnt leaves. Table 2. Demand of sugarcane leaves, and prices of sugarcane leaf purchased by biomass power industries in Khon Kaen Province, in the sugarcane production year of 2020/2021- 2021/2022. 3.2 Sugarcane leaf management and utilization by farmers The results of sugarcane leaves utilization by farmers in Table 3 shows that most farmers, who used a machine in fresh harvesting, had sold sugarcane leaves for additional income, covering 4,608.75 rai (1 rai = 0.16 ha) of the harvesting area following the recovery of sugarcane leaves in the field for weed control (3,788.25 rai) and the burnt leaves (1,056 rai). For manual fresh harvesting, most farmers had utilized the leaves for recovery of 673 rai sugarcane field. In the overview, 43.10 percent of total sample farmers sold sugarcane leaves in order to earn additional income. Furthermore, 40.30 percent of them had used sugarcane leaves through recovery of the sugarcane field as blanket for weed control. In addition, 98.60 percent of the 70 farmers had sold sugarcane leaves to local entrepreneurs, who had collected sugarcane leaves and made bale shape in the local areas (Table 4.). For customer channel, the local entrepreneurs had communicated with the key person, who had a harvesting machine, so they knew the farmers who would like to sell the sugarcane leaves from their fields.

224 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT Table 3. The results of sugarcane leaf utilization by farmers, classifying by harvesting methods. Table 4. Distribution channel for selling sugarcane leaves by farmers The selling price of sugarcane leaves that the farmers received was 2.84 USD per rai (Table 5.). According to the participants, the estimated selling price that they satisfy was 5.33 USD per rai. Without these prices, farmers could get the highest price of 28.42 USD per ton (in bale shape). To make the bale shape, they have to pay for sweeping and making square bale shape which cost around 11.65 USD per ton, including the cost of moving the bale shape to a truck of about 1.85 USD per ton. Then, they may also need to pay for transportation for 4.26 – 5.97 USD per ton, depending on the distance from a sugarcane field to a biomass power industry. Finally, selling sugarcane leaves per ton could offer net return for farmers about 8.95 to 10.66 USD per ton (Table 6.). Table 5. Average selling price of sugarcane leaves by farmers Table 6. Cost and return of sugarcane leaf management for selling in square bale shape with

225 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT The main reasons the famers sell the leaves were the easy sugarcane planted land management such as land preparation and improving soil fertility, the avoidance of burning the leaves, and following the government policy of no burning sugarcane leaves and straws” (Table 7.). Table 7. Reasons for selling sugarcane leaves of farmers. 4. CONCLUSION Sugarcane leaves serve as valuable materials for various purposes such as mulching, organic fertilizer, biomass power generation, biochar, and textile fiber. To ensure the sustainability of sugarcane leave utilizations, the research objectives are to estimate of the supply and demand of sugarcane leaves in Khon Kaen Province of Thailand, and to study the sugarcane leaf management and its utilizations by farmers. To achieve the objectives, three industries of biomass power generation and seventy farmers were interviewed face-to-face using the structured questionnaire. The findings conclude that only 51,500 tons of sugarcane leaves were supplied to the biomass power industry. There is still surplus supply of 282,279.58 tons in Khon Kaen. This suggests that consulting with the biomass power industry to enlarge demand is crucial. At the same time, encouraging farmers to utilize the sugarcane leaves is necessary to the concern. Without burning, recovery of and mulching the sugarcane leaves are alternative approaches for famer management. Additionally, selling sugarcane leaves also offers additional income for farmers. 5. ACKNOWLEDGMENT The authors would like to express appreciation for the support of the research fund from the National Research Council of Thailand. The authors also would like to acknowledge the Northeast Thailand Cane and Sugar Research Center for consultation. Research facility and office were supported by the Department of Agricultural Economics, Faculty of Agriculture, Khon Kaen University, Thailand. 6. REFERENCES Alves, M., G.H.S.F. Ponce, M.A. Silva and A.V. Ensinas. 2015. Surplus electricity production in sugarcane mills using rsidual bagasse and straw as fuel. Energy., 91: 751-757. Awe, G.O., J.M. Reichert and O.O. Wendroth. 2015. Temporal variability and covariance structure structures of soil temperature in a sugarcane field under different management practices in Southern Brazil. Soil & Tilage Reserch., 150: 93-106. Chandel, A.K., S.S. da Silva, W. Carvalho and O.V. Singh. 2012. Sugarcane bagasse and leaves: foreseeable biomass of bioful and bio-products. J. Chem. Technol. Bioechnol., 87: 11-20. Castioni, G.A., M.R. Cherubin, L.M.S. Menandro, G.M. Sanches, R.de O. Bordonal, L.C. Barbosa, H.C.J. Franco and J.L.N. Carvalho. 2018. Soil physical quality response to sugarcane straw removal in Brazil: a multi-approach assessment. Soil & Tillage Research., 184: 301-309. Ferreira-Leitão, V., L.M.F. Gottschalk, M.A. Ferrara, A.L. Nepomuceno, H.B.C. Molinari and E.P.S. Bon. 2010. Biomass residues in Brazil: availability and potential uses. Waste Biomass Valor., 1: 65-76. Go, A.W. and A.T. Conag. 2019. Utilizing sugarcane leaves/straws as source of bioenergy in Philippines: a case in the Visayas region. Renewable Energy., 132: 1230-1237. Jankaew, S. and K. Ariyakuare. 2021. Development of sustainable utilization processes from sugarcane leaves for the design of products design to the eco-efficiency. Retrieved from http:www.thai-explore.net/file_upload/ submitter/file_upload/2e4dacd0fcc48a8a7f8489da69a809c36e664178b2c69bde.pdf. Marin, F.R., P.J. Thorburn, L.G. da Costa and R. Otto. 2014. Simulating long-term effect of trash management on sugarcane yield for Brazilian cropping systems. Sugar Tech., 16(20): 164-173. Menandro, L.M.S., H. Cantarella, H.C.J. Franco, O.T. Kölln, M.T.B. Pimenta, G.M. Sanches, S.C. Rabelo and J.L.N. Carvalho. 2017. Comprehensive assessment of sugarcane straw: implications for biomass and bioenergy

226 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT production. Biofuels, Bioprod. Bioref., 11(2): 1-17. Office of Agricultural Economics. 2022. Agricultural Statistics of Thailand 2022. Retrieved from https://www.oae. go.th/assets/portals/1/ebookcategory/95_yearbook2565/. Office of The Cane and Sugar Board. 2021. Annual Report 2021. BangKok. Sampaio, I.L.M., T.F. Cardoso, N.R.D. Souza, M.D.B. Watanabe, D.J. Carvalho, A. Bonomi and T.L. Junqueira. 2019. Electricity production from sugarcane straw recovered through bale system: assessment of retrofit projects. BioEnergy Research., 12: 865-877. Kaewpradit, W. 2021. Sugarcane straw management to mitigate particulate matter and encourage sustainable sugarcane production. Khon Kaen Agriculture Journal., 49(1): 76-86. Yoddamnern, T. And S. Yotwinyuwong. 2020. Development of charcoal briquette from the cane leaves and bagasse to community enterprise. Journal of Energy and Environment Technology., 7(2): 12-24.

227 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT Chakrit Potchanasin*1 , Thanaporn Athipanyakul1 and Kuntonrat Davivongs1 1 Faculty of Economics, Kasetsart University, Bangkok 10900 Thailand. *Correspondence to: [email protected] ABSTRACT: This study aimed at investigating potential of bio-plastic as a co-products from cane and sugar industry. The study included (1) to reveal consumers’perception and willingnessto pay for bio-plastic, and (2) to investigate market value and potential of the bio-plastic. The study collected the data from interviewing the consumers in 4 main area of Thailand including Bangkok Metropolis, Chiang Mai, Khon Khean, and Songkhla during the year 2020 to 2021. Also, the study used descriptive analysis to describe consumers’ perception and attitude, production costs and returns, market value and commercial potential of the bio-plastic. Further, the study applied Contingent valuation double-bounded method to estimate willingness to pay of the consumers for the bio-plastic. The results had revealed that the consumers had less perception concerning the co-product which was presented by bio-plastic food box. However, the consumers attitude and awareness were in high level and also they had willingness to pay at double price for bio-plastic food box comparing to conventional plastic. In addition, the results showed positive potential of the bio-plastic indicated by increasing of market value from 11,548.81 billion baht in 2021 to 27,592.79 billion baht in 2025. In addition, global and domestic demand and willingness to pay for bio-plastic was growing. Also, the industry gained advantages from plenty of raw material and the special economic investment area to promote its production. Furthermore, producing bio-plastic would gain positive margin and profit and there would be supported in research and development from the government including various policiesto enhance the uses and production investment in bio-plastic industry. Based on the results, recommendations could be drawn that the government should provide more information of bio-plastic in orderto stimulate the people use.Also,supportsin investment and technological development from the government should be continually performed while incentive policies to enhance the use of bio-plastic was also needed. Moreover, improvement of regulations regarding especially definition of Granulated sugar and Factory would provide more processing alternatives instead of only sugar production which would benefit to bio-plastic industry. O-037 Potential of bio-plastic as an alternative co-product in sugarcane industry

228 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT 1. INTRODUCTION Changes in economic, social, and environmental factors can impact the sugarcane industry, leading to a decline in demand and economic value. To counteract this, the concept of developing sugarcane co-products aligns with the National 20-Year Strategy, which aims to drive the industry towards the future (New S curve). Bioplastics are one of the co-products that offer value-added potential for the sugarcane industry. Nevertheless, questions remain regarding the bioplastic industry's commercial potential and consumer reception. Thus, this research aims to study consumer awareness and willingness to pay for bioplastics from the sugarcane industry while evaluating the economic value and potential of bioplastic trade. The study results can guide the development of the bioplastic industry, serving as an alternative for co-production in the sugarcane industry's future development. Analysis used secondary and primary data collected by interviewing 2. RESEARCH METHODOLOGY DATA COLLECTION This research focuses specifically on bio-based plastic products of the PLA type because they are not yet widely used in the general market and have the highest usage proportion. The research explores consumer perception and willingness to pay for bio-plastic products in terms of PLA packaging for food products. Data were collected in four important areas - Bangkok, Chiang Mai, Khon Kaen, and Songkhla provinces - using a purposive sampling technique in 2020-2021. EMPIRICAL MODEL Tobit regression model was applied to investigate factors affecting the willingness to pay for a single-use transparent biodegradable plastic box with a size of 750 ml. The empirical model was explained asfollowing equation. WTPi = β0 + β1Gender_f + β2Age + β3eEdu + β4Occu_p + β5Inc + β6Reuse + β7Class + β8Search + β9Importance + β10Attempt + β11After_burn + β12Attd_1 + β13Attd_2 + β14Aware_1 + β15Aware_2 + β16BK + β17CM+ β18KK + ɛi WTPi = The willingness to pay for a single-use transparent biodegradable plastic box with a size of 750 ml. Gender_f = Gender (1 = male; 0 = otherwise) Age = Age of respondent (years) Edu = Number of years in school (years) Occu_p = Occupation of respondent (1 = working in private sector; 0 = otherwise) Inc = Income of respondent (THB/month) Reuse = Reusing plastic food containers (1 = reusing plastic food containers; 0 = otherwise) Class = Separate waste before disposal disposing of plastic food containers (1 = Separate waste before disposal; 0 = otherwise) Search = Choose to purchase biodegradable plastic food containers (1 = Choose; 0 = otherwise) Importance = Assessing the importance level of plastic types for plastic food containers (Rating on a scale of 1 to 10) Attempt = The level of effort in selecting and using biodegradable plastic for food containers (Rating on a scale of 1 to 10) After_burn = Behavior after using a food container (1 = burning; 0 = otherwise) Attd_1 = Attitude rating in the topic: The government should increase public awareness about biodegradable packaging. (Rating on a scale of 1 to 5) Attd_2 = Attitude rating in the topic: The government should create incentives for product manufacturers to use biodegradable plastic packaging (Rating on a scale of 1 to 5) Aware_1 = Awareness rating in the topic: In daily life, consumers should help each other reduce the amount of plastic usage. (Rating on a scale of 1 to 5) Aware_2 = Awareness rating in the topic: The level of the problem regarding the quantity of plastic packaging overall (Rating on a scale of 1 to 5) BK = Interview location in Bangkok (1 = Bangkok; 0 = otherwise) CM = Interview location in Chiang Mai province (1 = Chiang Mai province; 0 = otherwise) KK = Interview location in Bangkok (1 = Khon Kaen Province; 0 = otherwise) ɛi = Error term

229 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT The content analysis and quantitative analysis were conducted to calculate the product cost and return for each stakeholder along the value chain. Moreover, assessment of the trade values of the bioplastic industry was investigated. Estimation of the bioplastic value is based on primary data obtained from interviewing the bioplastic production company and other stakeholders regarding to its current and future market needs and prices. Furthermore, the analysis includes an analysis of the trade potential, which is conducted through a model known as Diamond's Model. The analysis considers four dimensions: (1) Objective conditions, (2) Factor conditions, (3) Related and supporting industries, and (4) Strategy, structure, and rivalry. Furthermore, it considers the role of the government and unpredictable or uncontrollable opportunities that impact competitiveness. 3. RESULTS Most of the sample were female (72.93%) with an average age of 34.29 years and a bachelor's degree as the highest level of education. Most were employed in private companies with an average monthly income of 26,816.27 baht, and an average household income of 73,315.07 baht per month. Consumers used clear single-use plastic food containers an average of 2.94 times per week, with an average of 1.87 boxes used per time. The average price of food per box was 55.43 baht. Most consumers (61.35%) purchased food in bioplastic containers from restaurants. Most of the food purchased was single-serve dishes, ready-to-eat meals, and snacks. After using the containers, 77.73% of consumers disposed of them, with 48.69% separating them for recycling. 66.16% of consumers did not consider the type of plastic used, whether it was conventional or biodegradable plastic, when making their food purchases. Consumers rated the importance of the type of plastic used in food packaging as moderately important, with an average score of 5.45 out of 10 points. The results revealed that important factors influencing the willingness to pay for a single-use transparent biodegradable plastic box include the occupation of consumers related to the private sector, consumer behavior regarding the reuse of plastic food containers, behavior in separating waste when disposing of plastic food containers, behavior in choosing to purchase biodegradable plastic food containers, behavior after using plastic containers, the level of awareness regarding the overall plastic packaging problem, interview location in Bangkok, and interview location in Khon Kaen. The willingness-to-pay value for biodegradable plastic containers is equal to 6,697 Baht per box, which is higher than the price of regular plastic containers by approximately 2 times, closely comparable to the price level comparison between biodegradable plastic and conventional plastic currently. It was found that the biodegradable plastics industry uses raw materials derived from raw sugar. The research results revealed that the production cost of PLA bioplastic pellets is 81.87 Baht per kilogram, while PBS bioplastic pellets have a production cost of 154.94 Baht per kilogram. In terms of profit, PBS bioplastic pellets generate a profit of 38.73 Baht per kilogram, which is higher than the profit of 20.47 Baht per kilogram for PLA bioplastic pellets. These prices are approximately 2.06 and 3.91 times higher than regular plastics, respectively. The analysis of the bioplastic production supply chain showed that the majority of profit, accounting for 97% of the total profit, comes from the factories that convert bioplastic pellets into bioplastic products. In contrast, farmers and sugar factories contribute only 3% to the total profit. Regarding market value, the market value of bioplastic pellets in 2021 amounts to 11,548.81 million Baht, with PLA bioplastic pellets accounting for 7,675.36 million Baht and PBS bioplastic pellets accounting for 3,873.44 million Baht. If the bioplastic pellets are further processed into bioplastic products, the market value increases to 24,826.88 million Baht. Trend analysis indicates that the market value of bioplastic products in 2024 is expected to increase to 27,592.79 million Baht, with the market value of PLA bioplastic products estimated at 17,909.18 million Baht and PBS bioplastic products estimated at 9,683.61 million Baht. If the bioplastic pellets are further processed into bioplastic products, the market value will reach 59,317.20 million Baht. In terms of trade potential, it has been observed that there is a growing demand for bioplastics both domestically and globally. Consumers show a willingness to pay a higher price for bioplastics. Thailand possesses favorable conditions for bioplastic production, including access to raw materials, investment incentives, and attractive profits for manufacturers. Furthermore, efforts are being made to develop biotechnology industries to support research and development. The bioplastic industry in Thailand experiences limited competition and operates within a market structure characterized by a small number of competitors, with 6-7 major players in the industry. The Thai government places significant importance on the development of the bio-based chemical industry and actively promotes the use of bioplastics while reducing and eliminating the use of single-use plastics. Measures such as tax incentives for companies that purchase and use bioplastic products and legislation to encourage investment in the bioplastic industry have been implemented. Additionally, the industry benefits from opportunities arising from the increased demand for food packaging due to the COVID-19 pandemic, as well as the growing trends in resource conservation and environmental preservation.

230 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT Furthermore, various international laws promote the use of bioplastics in different countries. 4. CONCLUSION AND RECOMMENDATION The conclusion and recommendation can be drawn from this research are five issues. The first recommendation is that consumers have limited knowledge and understanding of bioplastics. Therefore, the government should prioritize disseminating knowledge about bioplastics and their environmental benefits. This will contribute to increasing the demand for bioplastics, aligning with the potential of consumers who have a high level of awareness and consciousness regarding bioplastics. Secondly, it can be observed that many types of bioplastic products are still at the research stage, with limitations in terms of commercial development in the industry. Therefore, Thai government should promote the development of bioplastic products at the industrial level. Additionally, Thailand has technological limitations in production processes. Therefore, besides promoting research advancements to enable industrial production, the government should foster collaborations and partnerships with foreign technology owners through investment or joint ventures. This will help stimulate the development of the bioplastic industry within the country. Finally, In the case of promoting the use of sugarcane as a raw material, the government may need to consider amending various laws, including expanding the definitions of "sugarcane" and "factory" in Section 4, as well as modifying provisions and specifications in the Sugarcane and Sugar Act of 1984 related to the interpretation of products derived from sugarcane. 5. ACKNOWLEDGMENT I would like to express my gratitude to the National Research Office for their support in research funding. 6. REFERENCE National Strategy (BE 2561-2580). 2561. Royal Gazette: October 13, 2561.

231 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT Jiraporn Promkunthod1, Nattha Nitwatthanakul1, Sophon Wongkaew1 and Arak Tira-umphon1,2* 1School of Crop Production Technology, Institute of Agricultural Technology, Nakhon Ratchasima, 30000, Thailand 2Innovation of Quality Enhancement of Agricultural Products for Agro-industry-Research Center, Suranaree University of Technology, Nakhon Ratchasima, 30000, Thailand. *Correspondence to: 1 School of Crop Production Technology, Institute of Agricultural Technology, Nakhon Ratchasima, 30000, Thailand. [email protected] ABSTRACT: The developed medium and technique were further studied for economic factors affecting growth and development of sugarcane tissue under the low-cost tissue culture techniques including 1) culturing condition, 2) light source, 3) tissue culture medium grades, and 4) culturing containers, compared to that of the traditional system. The comparison was made at 2 growth stages; seedling multiplication and root induction. At the seedling multiplication stage, the open-system (un-autoclaved medium, room temperature with LED light source, lab-grade MS medium supplemented with sucrose and 15 mg/ml Sodium hypochlorite (NaOCl), and PE plastic containers) yielded 28.4 shoots/ container, 3.6 times higher than that of the traditional system which gave only 9 shoots/ container when assessed at 30 days after culturing. At the root induction stage; the number of roots, leaves, and stem height were not significantly different among plants cultured in the two systems; but the number of shoots developed under the open system (43 shoots/bag). When the production cost was estimated based on 10,000 seedlings, it amounted to 3.90 THB per seedling for the traditional system, whereas the cost for the open system was only 2.88 THB per seedling. This indicates a potential cost reduction of up to 1.35 times, save costs by 26.15%. Keywords: sugarcane, low-cost tissue culture, open tissue culture, NaOCl P-015 Study of economic factors affecting growth and development of sugarcane tissue under the low-cost tissue culture techniques

232 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT 1. INTRODUCTION Sugarcane (Saccharum officinarum L.) is an economically important plant in the sugarcane and sugar industry of Thailand. The major problems in sugarcane production include diseases transmitted through the planting material, such as sugarcane white leaf disease caused by sugarcane white leaf phytoplasma, sugarcane grassy shoot disease caused by sugarcane grassy shoot phytoplasma, red rot disease caused by Glomerella tucumanensis Raid, and smut disease caused by Ustilago scitaminea. Methods of preventing and controlling diseases carried by sugarcane planting materials include soaking the planting materials in hot water and using chemical agents for plant disease prevention and control. However, these methods have not been able to completely eliminate the pathogens that reside within the planting materials[1] In addition, it also causes a decrease in the germination percentage of the planting materials[2]. Currently, there is no available treatment and no sugarcane variety that is resistant to white leaf disease. Using planting materials free from disease pathogens is the most effective way to reduce the initial source of plant disease outbreaks in cultivation areas. Currently, sugarcane production using tissue culture technology to produce disease-free sugarcane varieties is an alternative option. However, it involves relatively high initial costs, especially for tools and equipment. There are expenses associated with maintaining and servicing the relevant equipment, as well as the need for skilled labor to carry out the operations [3]When considering it from a business or industrial perspective, the investment required is high compared to conventionalseedling propagation. The cost per unit may be too high to be practical[4]. Furthermore, there have been reports on the use of antimicrobial compounds (chemical, botanical, or a combination of both) in plant open tissue culture techniques. These reports suggest that using antimicrobial compounds instead of autoclaving to sterilize the media can reduce the cost per plantlet. This approach not only saves on electricity expenses but also allows the operation to be conducted in an open condition, thus preventing microbial contamination [5,6]. Plant open tissue culture technology refers to a method of plant tissue culture where the culture process is carried out in an open or semi-open environment, as opposed to a closed and sterile laboratory setting. It involves using antimicrobial compounds, both chemical and botanical, to control microbial contamination without relying on autoclaving for media sterilization. This approach offers potential benefits such as cost savings, increased efficiency, and reduced equipment requirements. By utilizing suitable concentrations of antimicrobials, plant open tissue culture technology aims to establish and maintain healthy cultures while minimizing the risk of contamination[5,7,8]. In order to produce a large quantity of sugarcane seedlings for the industry using open tissue culture technology, it is necessary to study certain factors to reduce production costs and lower the cost per seedling. Therefore, this research investigates the developed medium and technique were further studied for economic factors affecting growth and development of sugarcane tissue under the open-culturing conditions including 1) culturing condition, 2) light source, 3) tissue culture medium grades, and 4) culturing containers, compared to that of the traditional system. This approach can contribute to reducing the production costs of disease-free sugarcane seedlings. 2. MATERIALS AND METHODS Preparation of disease-free sugarcane seedlings To select the Khon Kaen 3 sugarcane variety cuttings, vigor and disease-free aged 8-10 months from the Khon Kaen Field Crops Research Center. Cut them into segments, each segment containing one bud. Soak the sugarcane segments in hot water at a temperature of 50°C for 2 hours. Then, plant them in a sand tray that has been sterilized with sprayed steam. Place the tray in a laboratory with fluorescent light. Close the lid of the tray and spray it with mist every 3 days to maintain moisture in the tray. After approximately 7-10 days, the sugarcane segments will sprout. When the sugarcane shoots reach 3-4 inches in length, cut the shoot tips containing actively growing tissue. These shoot tips will be used for closed system tissue culture to produce disease-free sugarcane seedlings as initial materials for experiments. Traditional system tissue culture, there are three stages of growth 1) Shoot Culture: This stage involves the initial development of shoots from the explants. Cut the youngest shoot tightly against the stem. Wash it thoroughly with running water. Peel off the outer sheath of the young shoot. Disinfect it by soaking in 70% ethanol for 1 minute, followed by 2% Clorox for 8 minutes. Transfer it to a culture cabinet. Rinse it three times with autoclaved distilled water to eliminate contaminants. Cut the growing tissue beneath the shoot apex under a stereo microscope. Culture the growing tissue in liquid medium, (Formula 1) on a shaker at a speed of 100 rotations per minute. Cultured for 6 weeks. 2) Shoot Multiplication: When the growing tissue develops into new shoots, it is transferred to a second medium (Formula 2) for further proliferation. This promotes the growth of new shoots (10 shoots per original tissue). The medium is changed every 2-3 weeks to increase the number of plantlets. 3) Root Induction: When a sufficient quantity of young shoots is obtained, they are transferred to a third medium (Formula 3) to induce root formation. They are kept for 4-5 weeks until sugarcane seedlings with well-developed roots are obtained. These seedlings are then ready to be transferred from the containers to the greenhouse for further cultivation. There are three stages of

233 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT growth-controlled environment with a temperature of 25+2 °C and a light intensity of 80-100 μmol/m2/s for 16 hours per day and 8 hours of darkness. Formula 1: MS (Murashige and Skoog, 1962) medium is prepared by adding growth regulators IBA at a concentration of 1 mg/L and GA at a concentration of 2 mg/L. Formula 2: MS medium is prepared by adding growth regulators KN at a concentration of 1 mg/L. Formula 3: MS medium without the addition of growth regulators. There are three formulas added at a concentration of Citric acid at 150 mg/L and sucrose at 20 g/L. The pH of the medium is adjusted to 5.6-5.8. The medium is then autoclaved at a temperature of 121 °C and a pressure of 15 pounds per square inch for 15 minutes. Method in the low-cost tissue culture techniques Extract the sugarcane tissue from the shoot multiplication stage of the traditional tissue culture system for further cultivation as an explant. The culture medium was prepared under open-culturing conditions outside an aseptic chamber. The medium for the shoot multiplication stage was prepared by adding growth regulators, such as 1 mg/L KN, 150 mg/Lcitric acid, 20 g/Lsucrose, and certain concentrations of 15 mg/ml NaOCl(replacing autoclaving).The tissue culture medium is then classified into separate grades: lab-grade MS medium and hydroponic-grade medium. After adjusting the pH to 5.6-5.8, both are packaged and sealed in two types of culturing containers: glass containers and PE plastic containers. The culturing containers are kept in a controlled environment with a temperature of 25+2°C and a non-controlled environment at room temperature. In the cultivation room, there are two types of lighting systems for tissue culture shelves: LED and fluorescent lights. These lighting systems provide a light intensity of 80-100 μmol/ m2/s for 16 hours per day, followed by 8 hours of darkness. After culturing at 30 days are transferred to root induction without the addition of growth regulators, 150 mg/L citric acid, 20 g/L sucrose, and certain concentrations of 15 mg/ml NaOCl (replacing autoclaving). In the sugarcane tissue cutting step, it is performed under open-culturing conditions outside an aseptic chamber. Begin by cleaning the bench with 70% alcohol. Dip the various tools in alcohol and then flame them. Set up an alcohol burner in the tissue transfer area. Experimental design The experiment is planned in a split-plot design. The main plot consists of FactorA: Room culturing environment with 2 levels: controlled room temperature at 25 °C and non-controlled room temperature. The sub-plot has a factorial arrangement of sub-plots involving three factors as follows: Factor B: light source with 2 levels: fluorescent light bulbs and LED light bulbs, Factor C: medium grade for tissue culture with 2 levels: lab-grade MS medium [9] and hydroponic-grade: SUT NS#5 medium [10], and Factor D: type of culturing bottle with 2 levels: glass bottles and plastic containers. It is divided into 16 treatments, each repeated 5 times. Divide it into two stages: the stage of increasing the number of shoot and the stage of root induction. Once suitable factors for open tissue culture are obtained, compare them with closed tissue culture. Data recording: Record the percentage of microbial contamination, browning, survival rate, and tiller emergence at 15, 30, and 45 days of culture. Data analysis: Analyze the ANOVA variance for each experiment and compare the means of each method using Duncan's New Multiple Range Test (DMRT) at a confidence level of 95%. 3. RESULTS AND DISCUSSION The developedmediumand techniquewere furtherstudied for economic factors affecting growth and development of sugarcane tissue under the low-cost tissue culture techniques, it was found that in the stage of shoot multiplication, there were no statistically significant differences (p>0.05) in the percentage of browning, contamination, and survival rate of sugarcane seedlings. The trends of browning percentage, contamination, and survival rate were consistent. However, in terms of number of shoots, there were statistically significant differences (p<0.01) among the different treatment combinations. After 15 and 30 days of tissue culture, the treatment combination of T10 (non-controlled temperature room, LED lights, lab-grade medium, plastic container) had the highest average number of shoots, which was 8.60 and 28.40 shoot per plastic container, respectively. (Table 1) In the root induction stage, each treatment combination had a statistically significant effect (p<0.01) on the number of roots and shoots of sugarcane seedlings. After 15 days of tissue culture, treatment combination T2 (controlled temperature room, LED lights, lab-grade medium, plastic container) had the highest average number of roots, which was 12.40. However, the average number of shoots was only 11.00 shoots per glass container, which was statistically different from treatment combination T10 (non-controlled temperature room, LED lights, lab-grade medium, plastic container) with an average of 6.00 roots but the highest average number of shoots of 43.00 shoots per bag. From the above experiment, it can be observed that the temperature of the culture room significantly influenced

234 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT the number of roots and tiller emergence. Additionally, the light source and type of container also had a significant effect on the number of roots and number of shoots of sugarcane seedlings. As for the medium grade, it did not affect the number of roots but had a statistically significant effect on the number of shoots of sugarcane seedlings (Table 2). From the above experiment, it can be observed that the temperature of the culture room significantly affects the number of roots and number of shoots, which is statistically significant. Sugarcane is classified as a C4 plant, and the optimal temperature for germination and sprouting of sugarcane stalks is between 32 and 38 °C. Growth slows down when the temperature drops below 25 °C, and it ceases when the temperature exceeds 38 °C. Temperatures above 38°C lead to a reduction in the rate of photosynthesis, but the respiration of sugarcane increases. [11] Therefore, from the experimental results, it can be observed that the cultivation of sugarcane tissue in a non-controlled temperature room (average temperature of 28-30 °C) resulted in the highest average number of shoots of 43.00 shoots. Consequently, it can be concluded that it is possible to cultivate sugarcane tissue in a non-controlled temperature room. According to the experiment, it can be seen that the light source and type of container have a statistically significant effect on the number of roots and number of shoots of sugarcane seedlings. When cultivating sugarcane tissue using white LED tubes with a light intensity of 80-100 µmol/m2/s, it wasfound to be more effective and resulted in a higher quantity of sugarcane compared to using fluorescent light sources. This finding is consistent with what researchers have reported regarding the effects of different intensities of white light from LED light sources on the growth of Khon Kaen 3 sugarcane tissue[12]. Therefore, for tissue culture propagation of sugarcane, it is recommended to use artificial light sources from LED light sources with a light intensity of 128 µmol/m2/s, as it resulted in the highest number of shoots. Furthermore, the type of container used for cultivation also influenced the number of shoots of sugarcane seedlings.It was observed that using plastic potsinstead of glass bottlesled to increased number ofshoots. Specifically, using transparent PP plastic pots, which are heat-resistant, durable, flexible, and have good impact resistance, proved to be more cost-effective compared to glass bottles. This makes them suitable for small-scale farmers or those with limited investment budgets starting their tissue culture propagation. Additionally, it was found that the grade of the culture medium did not affect the number of roots of sugarcane seedlings but had a statistically significant effect on number of shoots. The use of a higher-grade medium resulted in a greater quantity of sugarcane shoots compared to using a hydroponic-grade medium. This may be due to the fact that the commercially available hydroponic nutrient formulas are not yet suitable for sugarcane tissue culture. However, if the nutrient composition is adjusted to be similar to the higher-grade medium used in the experiment, it may be applicable for tissue culture propagation. This adjustment would further reduce the cost of tissue culture propagation. Table 1 The effect of economic factors on the growth and development of sugarcane tissue during the shoot multiplication stage under low-cost tissue culture techniques at 15 and 30 days. 1/ (**) The mean difference is highly significant at the 0.01 level., (ns)The mean non-significant. Note: TR = Temperature control room, NTR = non-temperature-controlled room, L = LED, F = Fluorescent, MS = MS medium, HP = Hydroponic grade, GB = Glass bottle, PP = PP plastic bag

235 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT From the tissue culture propagation experiment conducted using a traditional system (using autoclaving, temperature-controlled room, LED lights with MS-grade culture medium supplemented with sucrose, and tissue culture plants in glass bottles) compared to an open system (without using autoclaving, non-temperature-controlled room, LED lights with MS-grade culture medium supplemented with sucrose, and tissue culture plants in plastic pots with 15 mg/l sodium hypochlorite solution), as shown in Table 2, it was found that during the shoot multiplication stage, there was no statistically significant difference (p>0.05) in the percentage of browning, contamination, and survival rate between the closed and open system. However, during the shoot multiplication stage of the sugarcane seedlings after 15 days of propagation, a statistically significant difference was observed (p<0.05). Furthermore, after 30 days of propagation, the statistical significance became even more pronounced (p<0.01) in Table 3. In the open system, there were 28.4 shoot per pot, while in the traditional system, there were only 9 shoot per bottle after 30 days of propagation. In terms of root induction, number of leaves, plant height, and contamination, there was no statistically significant difference between the open and closed system. However, a higher number of shoot was observed in the open system, with 43 shoot per bag (Table 4 and Fig.1) Table 2 The effect of economic factors on the growth and development of sugarcane tissue during the root induction stage under low-cost tissue culture techniques at 15 days. 1/ (**) The mean difference is highly significant at the 0.01 level., (ns)The mean non significant. Note: TR = Temperature control room, NTR = non-temperature-controlled room, L = LED, F = Fluorescent, MS = MS medium, HP = Hydroponic grade, GB = Glass bottle, PP = PP plastic bag Table 3 The effect of the plant tissue culture method on the growth and development of sugarcane tissue during the shoot multiplication stage at 15 and 30 days. 1/ (*) The mean difference is significant at the .05 level., (**) The mean difference is highly significant at the 0.01 level., (ns)The mean non-significant.

236 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT Table 4 The effect of the plant tissue culture method on the growth and development of sugarcane tissue during the root induction stage at 15 days. 1/ (*) The mean difference is significant at the .05 level., (**) The mean difference is highly significant at the 0.01 level., (ns)The mean non-significant. Figure 1 Sugarcane plantlets were produced using conventional tissue culture methods (A, B) and low-cost open tissue culture techniques (C, D). A and C represent the shoot multiplication stage at 30 days, while B and D represent the root induction stage at 15 days. Comparing production cost of tissue culture propagation between traditional system and open system, low-cost tissue culture techniques. The cultivation of sugarcane tissue culture using an open system approach incurs lower costs due to various factors. These factorsinclude the use of non-temperature-controlled cultivation rooms, PE plastic containersinstead of glass bottles, LED lightsinstead of fluorescent lights, and the utilization of sodium hypochlorite or hydrogen peroxide instead of high-pressure steam sterilization, among others. When calculating the cost of tissue culture propagation, it was found that for the cultivation of 10,000 tissue culture samples, the closed-system approach (8-ounce glass bottles) resulted in a high cost of 38,969.25 Baht. In contrast, the open-system approach (12-ounce plastic containers) incurred a lower cost of only 28,793.59 Baht (Table 5). When calculating the cost per plantlet, it was determined that the closed-system approach had a cost of 3.9 Baht per plantlet, whereas the open-system approach had a lower cost of 2.88 Baht per plantlet. This translates to a cost reduction of up to 1.02 Baht per sample, saved 26.15% of the cost for 10,000 sugarcane plantlets. In summary, the open-system approach can reduce production costs by up to 1.35 times compared to the closed-system approach. Our results were consistent with those reported in studies indicating that the presence of a Qianxing No.1 in non-autoclaved media for sugarcane micropropagation it was considerably reduced by 0.4 Yuan per plantlet, saving cost 40% compared to that of the traditional method, 1.0 Yuan per plantlet if the open tissue culture technology was used in the production of 10,000 sugarcane plantlets. [13] However, the research team believes that it might be possible to further reduce the production costs throughout the sugarcane tissue culture production chain for commercial seedling production (costs in terms of labor and time). This could include using hydroponic-grade tissue culture media instead of laboratory-grade media and implementing a vertical seedling nursery system, among other strategies.

237 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT Table 5 The cost of production of plantlets by two tissue culture techniques. 1/Based on recurring expenses incurred on various items for producing a batch of 10,000 sugarcane plantlets in 2 month. 2/Electricity bills are not included. 4. CONCLUSIONS A study of certain factors to reduce production costs that affect the growth and development of sugarcane tissue culture in an open system found that during the seedling multiplication phase, the open system had 28.4 shoot per pot, while the closed system had only 9 shoot per bottle. The efficiency of tiller emergence during the seedling multiplication phase in the open system was 3.16 times higher than the closed system. After 30 days of propagation, there was no statistically significant difference in the number of roots, number of leaf, plant height, and contamination between the open and closed systems. However, there was a significant difference in tiller emergence, with the open system having 43 shoot per bag, while the closed system had only 19.4 shoot per bottle. The efficiency of tiller emergence during the root induction phase in the open system was 2.22 times higher than the closed system. When the production cost was estimated, based on 10,000 seedlings, it was 3.90 THB/ seedling for the traditional system while that of the open system was only 2.88 THB/seedling. Low-cost tissue culture techniques save costs by 26.15% compared to the traditional method. 5. ACKNOWLEDGEMENT This research is supported by National Research Office, Khon Kaen Field Crops Research Center, Innovation of Quality Enhancement of Agricultural Products for Agro-industry-Research Center form Suranaree University of Technology, Thailand for their support in providing personnel, experimental facilities, and related scientific instruments. 6. REFERENCES [1] Kaewpratumrussamee, C., Koohapitakthum, R., Chatchawankanpanich, O., Chachiyo, H. and Wongcharoen A. (2015). A suitable culture medium for micropropagation of sugarcane (Saccharum officinarum L.) in vitro. Khon Kaen agr. J. 43 (suppl.1), pp. 170-175. https://ag2.kku.ac.th/kaj/PDF.cfm?filename=O028 Pat_08.pdf&id=1835&keeptrack=10 [2] Kaewmenee, C., and Y. Hanboonsong. (2011). Evaluation of the efficiency of various treatments used for sugarcane white leaf phytoplasma control. Bulletin of insectology. 64, pp. 197-198. http://www. bulletinofinsectology.org/pdfarticles/vol64-2011-S197-S198kaewmanee.pdf [3] Ahloowalia, B and Savangikar, V. (2002). Low cost options for energy and labour. In Proceedings of a Technical Meeting organized by the Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture and held, pp 41-44. August 26–30, 2002, Vienna, Austria. http://www-pub.iaea.org/MTCD/publications/PDF/te_1384_web.pdf [4] George, P and Manuel, J. (2013). Low cost tissue culture technology for the regeneration of some economically important plants for developing countries. International Journal of Agriculture, Environment and Biotechnology. 6(suppl.), pp. 703-711. https://www.researchgate.net/publication/305688749_Low_cost_tissue_ culture_technology_for_the_regeneration_of_some_economically_important_plants_for_developing_countries [5] Sawant, R. A and Tawar, P. N. (2011). Use of sodium hypochlorite as media sterilant in sugarcane micropropagation at commercial scale. Sugar Technology. 13 (1), pp. 27-35. DOI:10.1007/s12355-011-0072-6

238 The 2nd International Conference on Cane and Sugar 2023 Towards BCG Economy; Smart Farm to Bio Industry ECONOMIC AND MANAGEMENT [6] Lu, J., Tira-umphon A., Zhang, Z., Lei, S. and Yu, L. (2014). Effect of bacteriostat (Qianxing No.1) tissue culture of sugarcane. Molecular Biology and Tissue Culture. 15(9), pp. 1478-1481. https://www. researchgate.net/profile/Arak_Tira-Umphon/publication/295812663_Effect_of_BacteriostatQianxing_No1_ on_Open_Tissue_Culture_of_Sugarcane/links/56cd8a6e08ae059e37531b9f/Effect-of-BacteriostatQianxing-No1-on-Open-Tissue-Culture-of-Sugarcane.pdf [7] Cui, G., Wenxiu, D., Xu, Q and Zhongxu, S. (2004). The preliminary study on plant open tissue culture. Journal of Shandong Agricultural University (Natural Science). 35(4), pp. 529-533. [8] Lu, J., Ali, A., He, E., Yan, G., Tira-umphon A. and Gao, S. (2019). Establishment of an open, sugar-free tissue culture system for sugarcane micropropagation. Sugar Tech. 22(8). DOI:10.1007/s12355-019-00758-1 [9] Murashige, T. and Skoog, F. (1962). A revised medium for rapid growth and bioassays with tobacco cultures. Physiol. Plant. 15, pp. 473-497. [10] Tira-umphon A. (2015). Hydroponics. Nakhon Ratchasima: School of Crop Production Technology, Institute of Agricultural Technology, Suranaree university of technology. Retrieved June 13, 2023, from http://sutir.sut. ac.th:8080/jspui/handle/123456789/7650 [11] Ashok k. Srivastava1, A. and Rai, M. (2012). Review: Sugarcane production: Impact of climate change and its mitigation. Biodiversitas. 13(4), pp. 214-227. DOI: 10.13057/biodiv/d130408 [12] Thanyasiriwat, T., Praneetpolkrang, T. and Kawicha, P. (2020). Effect of led light intensities on growth of sugarcane cv. Khon Kaen 3 shoots cultured intemporary immersion bioreactor system. Thai Journal of Science and Technology. 9(6), pp. 821-831. https://www.researchgate.net/publication/343569555_Effect_of_LED_ Light_Intensities_on_Growth_of_Sugarcane_cv_Khon_Kaen_3_Shoots_Cultured_in_Temporary_Immersion_ Bioreactor_System [13] Lu, J. (2014). Development of low-cost open tissue culture technology for sugarcane. Degree of Doctor of Philosophy in Crop Science. Suranaree University of Technology. http://sutir.sut.ac.th:8080/jspui/ handle/123456789/8120

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