BANGLADESH JOURNAL OF SUGARCANE
Volume 35 June 2014
BANGLADESH SUGARCROP RESEARCH INSTITUTE
ISHURDI-6620, PABNA, BANGLADESH
BANGLADESH JOURNAL OF SUGARCANE
Volume 35 June 2014
EDITORIAL BOARD - Editor
- Associate Editor
M. Khalilur Rahman, M.S. & Ph.D. (USSR) - Assistant Editor
Md. Amzad Hossain, M. Sc. (Ag) (BAU), Ph.D. (Japan) - Member
Md. Anisur Rahman, M.S. (BAU), Ph.D. (China) - Member
A. S. M. Amanullah, M.S. & Ph.D. (BAU) - Member
Samajit Kumar Pal, M. Sc. (Ag) (BAU), Ph.D. (Philippines) - Member
Md. Shamsur Rahman, M. Sc. (Ag), Ph.D. (Thailand) - Member
Kuasha Mahmud, M.S. & Ph.D. (BAU) - Member
Md. Ataur Rahman, M.S. (BAU), Ph.D. (BSMRAU) - Member
Kohinur Begum, M.S. & Ph.D. (BAU) - Member
K. M. Rezaul Karim, M.S. (BAU) - Member
A. K. M. Rashadul Islam, M.S. (BAU) - Member
Md. Abu Taher Sohel, M.S. (BAU) - Member
Khandakar Mohiul Alam, M.S. (BAU), Ph.D. (China)
Md. Nur Alam Miah, M.S. (BAU)
ii
EDITOR’S NOTE
It is my pleasure to publish the 35th volume of Bangladesh Journal of
Sugarcane. Publication is one of the best ways to disseminate the research
findings around the globe. We are very happy to see our scientists are sharing
knowledge and communicating themselves the best possible way they can.
Research and development of ancillary sugarcrops along with sugarcane is a very
healthy sign for increasing sugar production in Bangladesh.
The present volume includes research results on performance of
sugarbeet varieties under agro-climatic conditions and fertilizer requirements. In
addition, results of different experiments conducted by our scientists like genetic
variability and diversity analysis, different intercrops, tissue culture, planting
materials, loss assessment of stem borer infestation, amendment of light textured
soil, phytoclarificant for goor production, flood stress, on farm evaluation and
screening against smut disease of sugarcane were furnished in this volume.
Scientists and related personnel from home and abroad will be able to get recent
information about sugarcrops research in Bangladesh from this issue.
The personnel of sugarcane and other ancillary crops will hopefully be
benefitted from the scientific articles published in this volume in context of
improving sugarcrops cultivation techniques as well as increase quality and yield.
The information contains in this issue will certainly bring some measurable
changes in upgrading the knowledge of those who are directly or indirectly
involved in the research and development of sugarcrops. Moreover, I firmly believe
that the articles of this volume would help to minimize field oriented problems
related to sugarcrops production and management. It is inevitable to find out ways
and means for ensuring time befitting quality research to face the future
challenges due to climatic change.
All those who spent their valuable time for the publication of this issue
deserve special appreciation and thanks.
Dr. M. Khalilur Rahman
Director General
BANGLADESH JOURNAL OF SUGARCANE
Volume 35
June 2014
Publication No. 205
Published by : Bangladesh Sugarcrop Research Institute
Ishurdi-6620, Pabna, Bangladesh
Web: www.bsri.gov.bd
Fax: 8807326-63888
iv
CONTENTS
Performance of Some Exotic Tropical Sugarbeet Genotypes under Agro-climatic
Conditions of Bangladesh
A.K.M.R. Islam, M.A. Razzak, M.J. Alam, M.A.T. Sohel, H.P. Roy, M.S. Rahman and M.K.
Rahman .......................................................................................................................... 1
Effect of Nitrogen and Potassium on Growth, Development and Sugar Accumulation
in Tropical Sugarbeet
M.N. Kashem, Q.A. Khaliq, M.A. Karim, A.J.M.S. Karim, S.M.R. Karim and
M.M. Hossain .................................................................................................................. 10
Germination and Yield of Tropical Sugarbeet as Influenced by Variety and Fertilizer
S. Islam, M.A.T. Sohel, M.A. Haque, M.A. Razzak, M.J. Islam, S.M.R. Karim, M.S. Islam
and K.M. Alam ............................................................................................................... 20
Genetic Variability, Correlation and Path Analysis of Some Yield Components of
Sugarcane Genotypes
R. Alam, M.A. Rahman, K.M.R. Karim, H.M. Tarique and A.C. Deb ............................. 28
Genetic Diversity Analysis and DNA Fingerprinting of Ten Sugarcane Germplasm
Using SSR Markers
A.K. Ghose, M.K. Hasan, K. Mahmud, N. Islam, M.A. Rahman, M.S. Arefin and M.A.
Hossain ......................................................................................................................... 37
Tolerance Mechanism of Some Sugarcane Genotypes under Flood Stress
M.K. Begum, M.S. Arefin and M.J. Islam ..................................................................... 48
Productivity and Profitability of Onion Seed Crop-Mungbean Sequential Intercrop-
ping with Sugarcane
M.J. Alam, M. M. Rahman, A.K.M.R. Islam, M.S. Hossain, M.A. Razzak, M.S Rahman,
H. P. Roy and S. Islam .................................................................................................. 60
Response of 2, 4-D on Callus Induction and Plantlet Regeneration in Sugarcane
K. Mahmud, K.M. Nasiruddin, M.A. Hossain. M.A. Rahman and A.K. Ghose ....... 73
Site Specific Fertilizer Requirement of Sugarcane and Potato as Intercrop under
Sugarcane Based Cropping System
S. Islam, M.A. Haque, M.S. Islam, M.A. Razzak, M.J. Islam, S.M.R. Karim and
K.M. Alam ...................................................................................................................... 79
(Continued)
v
CONTENTS
Effects of Phyto Clarificant as Powder Form on Goor Manufacture
M.S. Arefin, M.K. Begum, M.J. Islam, M.A. Rahman, A.K. Ghose and S. Islam ............ 88
Amelioration of Light Textured Soil through Addition of Ash and Press Mud for
Increasing Sugarcane Production
S.S. Tabriz, A.S.M. Amanullah, M.S. Hossen and M. A. Rahman ................................ 96
Paired Row System of Sugarcane Cultivation with Chilli as Intercrop under Tista
Meander Flood Plain Soils
S.M.R. Karim, M.A. Razzak, M.N. Kashem, S. Islam and M.H. Rahman ........................ 102
Loss Assessment of Sugarcane due to Attack of Stem Borer
M.A. Rahman, M.Z. Alam, M.R.U. Miah, M.E. Reza and M.N.A. Siddiquee ................ 108
Comparative Performance of Different Planting Materials of Sugarcane at Farmers
Field in High Ganges River Flood Plain Soils
M. A. Razzak, M.A.T. Sohel, A.K.M.R. Islam, M. Kamruzzaman, M.J. Alam, H. P. Roy and
M.S. Rahman ............................................................................................................... 118
On-farm Evaluation of Promising Sugarcane Clones in Different Agro-Ecological
Zones
M.L. Kabir, M.A. Rahman, M.H. Rahman, H.M. Al-Amin, H.P. Roy, M.I. Hossain, and
M.R. Islam .................................................................................................................... 124
An Empirical Study on Technical Efficiency of Sugarcane Production in Rajshahi
District of Bangladesh
S. Khatun, M. Kamruzzaman, M.S. Islam and M.H. Rahman ...................................... 131
SHORT COMMUNICATION
Screening of Some Sugarcane Genotypes Against Smut Caused by Ustilago
scitaminea Sydow
M.J. Uddin, M.S. Rahman, M.I. Hossain and M.O. Khaiyam ........................................ 138
vi
Bangladesh J. Sugarcane, 35 : 1-9 June, 2014
Performance of Some Exotic Tropical Sugarbeet
Genotypes under Agro-climatic Conditions of Bangladesh
A.K.M.R. Islam*, M.A. Razzak, M.J. Alam, M.A.T. Sohel, H.P. Roy, M.S. Rahman and
M.K. Rahman
Agronomy and Farming Systems Division
Bangladesh Sugarcrop Research Institute, Ishurdi-6620, Pabna, Bangladesh.
ABSTRACT
Sugarbeet is an important sugar producing crop in the world.
Generally sugarbeet is grown in temperate regions but now-a-days some
genotypes were introduced which can grow in tropical and subtropical regions
in the world which is called tropical sugarbeet variety. As sugarbeet is a short
duration (150-180 days) crop, so if we can introduce sugarbeet in Bangladesh,
sugar production will be increased with a great extent. Before introducing
sugarbeet we have to select suitable genotypes of sugarbeet which can grow
well in Bangladesh agro-climatic conditions. A comparative study was
conducted to assess the performance of 11 (eleven) exotic tropical sugarbeet
genotypes viz.,Cauvery, Shubrha, HI 0044, HI 0473, SZ 35, PAC 60008, SV1,
Danicia, Aranka, Sereneda, Natura at Bangladesh Sugarcrop Research
Institute (BSRI) farm, Pabna during the rabi season of 2013-2014 following
randomized complete block design with three replications. Sugarbeet
genotypes showed different behavior with respect to size, shape, beet yield
and brix % of beet. Among the genotypes of sugarbeet the highest yield (98.80
tha-1), brix % (18.48) and pol% (10.08) obtained from the genotype Aranka and
the lowest from PAC 60008 (72.94 tha-1). But in case of brix % and pol% of
beet all other cultivar were shown statistically similar. The overall results
revealed that the tropical sugarbeet genotypes Aranka, Cauvery, SZ 35,
Danicia, Sereneda, Natura can be grown successfully in Bangladesh.
Key words: Tropical sugarbeet, pol %, beet yield
INTRODUCTION
Sugarbeet (Beta vulgarisomit L.) a member of chenopodiaceous family is the
second important sugarcrop after sugarcane, producing about 40% of sugar annually all
over the world (Amr and Gaffer, 2010). It is a fleshy root crop processed for sugar
production. It is native to temperate countries and hence has been associated with the
temperate environment. The leading sugarbeet producing region includes the European
Union, the USA and Russia. Despite being a temperate crop, sugarbeet trials have been
going on in some selected tropical countries like India, Pakistan, Sudan and South Africa.
Kapur and Kanwar (1990) noted that sugarbeet can be grown successfully as a winter
crop in subtropical. Asadi (2007) reported that in some tropical and subtropical regions
like Sudan and Pakistan, sugarbeet processing can be done from 270 to 300 days per
year. Sugarbeet with its relatively short season can be accommodated in the crop
rotation of large agricultural schemes such as Gezira in Sudan. Proponents of tropical
sugarbeet production argue that this would be a viable solution to problems facing
* Corresponding author: A.K.M.R. Islam, Senior Scientific Officer
e-mail: [email protected]
2 Bangladesh J. Sugarcane, 35 : 1-9 June, 2014
sugarcane farmers in the tropics. According to David and Young (1981) sugarbeet
matures and is ready for harvest in 5-6 months. The short maturity period of sugarbeet
may result in a quick and reliable income for farmers like Bangladesh. Sugarbeet roots
contain high concentration of sucrose (Memon et al., 2004).
The yield and economic benefit from sugarbeet will be higher than any cereal
crop. Moreover, realizing its high sugar content, short growing season, sugarbeet as a
cash crop, will improve the income of farmers. For these reasons, intensive field tests on
sugarbeet in Bangladesh have been continued unabated at the Bangladesh Sugarcrop
Research Institute since early 2002 in an attempt to introduce sugarbeet in the sugar
industry in Bangladesh beside sugarcane. The sugarbeet is particularly well adapted in
irrigated agriculture (Follet et al., 1964). Sugarbeet has no self regulatory mechanisms to
promote sucrose accumulation but is dependent upon external stimuli from the climatic
factors such as light, temperature and day length which determine to a great extent, the
type of growth and the amount of sugar that gets stored in the root (Ulrich, 1952;
Petkeviciene, 2009). All tropical sugarbeet genotypes are preferable to evaluate them
under the agro-climatic conditions of Bangladesh to select the best suited ones. Osman
et al. (2003) found significant differences in the studied sugarbeet varieties in different
sources like root length, diameter, fresh weight, root and sugar yield (ton/fed), as well as
sucrose and purity percent etc. Bangladesh requires about 2 million tons of sugar per
year (FAO recommendation 13 kg sugar and 17 kg goor per person per annum) but the
present domestic production is about 0.6 million tons (0.1 million ton sugar and 0.5 million
tons jaggery) (Rahman et al., 2006). For fulfilling the demand of sugar, an alternate new
short duration sugar producing crop named sugarbeet has been introduced in Bangladsh.
Field trials of this crop have been continued at Bangladesh Sugarcrop Research Institute.
The results of these efforts were quite promising and in most cases very encouraging.
For this reason an attempt has been made to evaluate the performance of exotic tropical
sugarbeet genotypes under selected agro-climatic conditions of Bangladesh.
MATERIALS AND METHODS
The experiment was conducted at Bangladesh Sugarcrop Research Institute
farm, Ishurdi, Pabna during the rabi season of 2013-2014. The trial consisted of 11
(eleven) genotypes of sugarbeet viz., Cauvery, Shubrha, HI 0044, HI 0473, SZ 35, PAC
60008, SV-1, Danicia, Aranka, Sereneda, Natura which were supplied by Syngenta (BD)
Ltd.; SesvanderHave (Belgium) and KWS (Germany). The experiment was laid out in
Randomized Complete Block Design (RCBD) with three replications. Individual plot size
was 6m × 6m. The experiment was located in High Ganges River Flood Plain soils under
Agro-ecological Zone-11 with medium high land of typical sandy loam soil having pH 7.6.
The soil of the field was non-saline, non-sodic, and alkaline in reaction and had low
organic matter (OM), phosphorus (P) and nitrogen (N) contents. The experiment was set
in the month of November 2013. All recommended agronomic and cultural operations
including weeding, fertilization, irrigation and plant protection measures were followed
during the entire course of study on a standardized uniform pattern for all the plots. After
seed bed preparation Urea, TSP, MoP, Gypsum, Zinc sulphate and Boric acid were
applied at the rate of 260 Kg, 120 Kg, 225 Kg, 100 Kg, 10 Kg and 20 Kg per hectare,
respectively (Anonymous, 2005). Total TSP, Gypsum, Zinc sulphate, Boric acid, 1/3 Urea
and MoP was applied in the line before ridge preparation. Rest amount of Urea and MoP
Performance of Some Exotic Tropical Sugarbeet ......... Bangladesh 3
was applied in two installments at 30 and 60 days after sowing. Then the plots were
prepared and layout was made. Manual sowing of seed of sugarbeet varieties was carried
out and seeds were sown on light ridge soil at 3-4 cm depth. The distance between two
ridges was 50 cm and seed to seed distance was 20 cm. Management practices, like
thinning for once at 4-5 leaf stage, harrowing, weeding (3 times at 25, 50 and 75 DAS),
irrigation (5 times at 1, 15, 30, 60 and 90 DAS) and other intercultural operations were done
regularly. To control jute hairy caterpillar, soon after infestation, the larvae were destroyed
by hand. To control Spodoptera litura (a Lepidopterous insect), an integrated approach
was rendered. Nitro 505 EC (Chlorpyriphos + Cypermethrin) @ 0.1% was sprayed at 7
days interval. Sex pheromone trap (Spodoptera pheromone) was also placed @ 45 traps
per hectare in the field to male insect. Predator insect (Bracon hebetor) was also released
in the field to control Spodoptera litura. Ten plants were randomly harvested from each
replication at 165 (five and half months) days after sowing to determine yield and yield
attributes. Generated data were compiled and analyzed properly for interpretation of the
results. Data regarding germination percentage, plant height (cm), beet length (cm), beet
girth (cm), beet weight (gm), beet yield (tha-1) and beet brix % were collected. Germination
(%) was recorded at thirty days after planting while plant height (cm), beet length (cm), beet
girth (cm), beet weight (gm), beet yield (tha-1) and beet brix % were recorded at the time
of harvest. Brix was measured with the help of hand refractometer.
AB
CD
4 Bangladesh J. Sugarcane, 35 : 1-9 June, 2014
EF
GH
IJ
K
Figure 1. Picture of eleven exotic tropical sugarbeet genotypes (A = Cauvery, B = Shubrha,
C = HI 0044, D = HI 0473, E = SZ 35, F = PAC 60008, G = SV1, H = Danicia,
I = Aranka, J = Serenada and K = Natura)
Performance of Some Exotic Tropical Sugarbeet ......... Bangladesh 5
RESULTS AND DISCUSSION
A well adapted plant does not show any sign of adverse effect on its life cycle at
any stage of growth in new environment. Different yield attributing data such as
germination (%), plant height (cm), beet length (cm), beet girth (cm), beet weight (gm),
beet yield (tha-1) and beet brix % are presented in Table 1.
Germination percentage
All genotypes of exotic tropical sugarbeet showed a good germination
percentage. It was seen from the table 1 that germination percent among the genotypes
varied significantly. The highest germination percent was found in variety PAC 60008
(93.14%) which is statistically similar with SV1 and Cauvery (92.83 and 92.85%),
respectively which is similar with the genotypes SV1, Cauvery, Serenada, Natura, HI per
control 0473, Danicia. The lowest germination was found in Shubrha (81.52 %).
Table 1. Growth and yield performance of different exotic Sugarbeet genotypes
during rabi season of 2013-14 at BSRI farm, Ishurd, Pabna
Genotypes Germination Plant Beet length Beet girth Beet yield
(%) height (cm) (cm) (cm) (tha-1)
Cauvery 63.14 a-c 31.24
Shubrha 92.85 a 62.94 a-c 27.66 31.71 a-c 72.94 c-e
HI 0044 81.52 d 30.44 31.2 a-c 59.90 e
HI 0473 88.52 bc 67.18 a 33.48 ab 69.01 b-d
SZ 35 27.28
PAC 60008 91.98 ab 66.00 ab 30.30 30.08 bc 65.43 de
SV1 86.53 c 65.19 ab 27.67 29.70 c 76.61 cd
Danicia 93.14 a 62.04 bc 29.58 30.99 a-c 72.94 c-e
Aranka 92.83 a 69.79 a 29.01 31.54 a-c 82.64 a-c
Serenada 90.01 a-c 65.07 ab 27.20 32.64 a-c 94.26 ab
Natura 88.33 bc 66.37 ab 27.18 34.46 a 98.80 a
LSD (0.05) 90.77 ab 61.97 bc 30.54 32.0 a-c 85.76 a-c
91.72 ab 61.78 bc 30.89 a-c 85.91 a-c
NS
4.00 4.95 3.75 16.63
In a column, figures with similar letters do not differ significantly at 5% level
Plant height (cm)
Results in table 1 showed that the plant height varies significantly in different
tropical sugarbeet genotypes. The highest plant height was found in SV1 (69.79 cm) and
HI-0044 (67.18 cm) which was similar with the variety HI-0473, SZ-35, Danicia and
Aranka. Variety Natura, Serenada and Natura 61.78% showed lowest plant height. The
differences could be attributed to the genetic structure of the evaluated sugarbeet
varieties. These results were similar to those obtained by Aly (2006), El-Bakary (2006)
and Ismail et al. (2006).
6 Bangladesh J. Sugarcane, 35 : 1-9 June, 2014
Beet length (cm)
There were no significant differences in case of beet length among the tested
tropical sugarbeet genotypes (Table 1).
Beet girth (cm)
Beet girth was significantly affected in different tropical sugarbeet genotypes.
Maximum beet girth was found in variety Aranka (34.46 cm) which is similar with other
varieties except SZ-35 (Table 1). These results were similar to those obtained by Aly
(2006), El-Bakary (2006) and Ismail et al. (2006).
108 106 105
106
103.33 103.33
104 98 99
Number of leaves 102 101.67
100 98 99.33 99.67
98 95.67
96
94
92
90
Tropical sugarbeet genotypes
Figure 2. Number of leaves/plant of different exotic sugarbeet genotypes
Number of leaves per plant
Figure 2 indicated that there were significant differences in producing leaf during beet
growth periods among the genotypes. The highest number of leaves was found in
genotype PAC 60008 (106.00) and the lowest in SZ-35 (95.67). Rest of the genotypes
was produced more or less number of leaves.
Unit beet weight (g)
Unit beet weight is an important parameter of sugarbeet as it is directly related to
the yield per unit area. If unit beet weight is higher then yield will be higher. From figure 3
it was clear that unit beet weight is different among the tested tropical sugarbeet
genotypes. The highest unit beet weight (995.70 g) was found in the genotype Cauvery
and Aranka (992.76 g). The lowest beet weight was found in the genotype Shubrha
(835.95 g) followed by Natura (840.96 g).
Performance of Some Exotic Tropical Sugarbeet ......... Bangladesh 7
1050 995.7 992.76
1000
Unit beet weight (g) 954.26 968.05
950 933.64
900
850 852.38 870.7 877.9 893.17
840.96
835.95
800
750
Tropical sugarbeet genotypes
Figure 3. Unit beet weight of different exotic tropical sugarbeet genotypes
Beet yield (tha-1)
Results in table 1 indicated that tested tropical sugarbeet varieties showed
significant differences in beet yield. Among eleven genotypes of sugarbeet Aranka gave
the highest beet yield (98.80 tha-1) followed by Danicia (94.26 tha-1), Natura (85.91 tha-1),
SSheurebnharada(5(98.59.076thtah-1a)-.1)ThaendinScrVe1as(8e2i.n64roothtay-i1e)ldgewnaostyspterosn. gTlyherelloawteedsttoyrioeoldt was found in
performance,
i.e., root length, diameter and fresh weight. Similar results were obtained by Aly (2006),
El-Bakary (2006) and Ismail et al. (2006).
Sugar percent (Brix and pol % beet)
Regarding sugar contents, a significant difference was found among varieties for
brix and pol% (Figure 4 ). The highest brix percentage was observed in Subhra, Cauvery
Danicia and Aranka (18.48-18.87%) followed by SZ 35, PAC 60008, SV1, Sereneda and
Natura (17.88- 18.22 %), while the lowest in HI-0044 (17.39 %). Concerning pol percent
in sugarbeet extract, variety Aranka had the highest p ol%.
8 Bangladesh J. Sugarcane, 35 : 1-9 June, 2014
Sugar % 20 Beet Brix (%) Beet pol (%)
15
10
5
0
Tropical sugarbeet genotypes
Figure 4. Brix (%) and pol (%) of beet obtained by different exotic sugarbeet
genotypes
From the above results, it may be concluded that tropical sugarbeet cultivation is
possible in Bangladesh. Among the eleven genotypes of exotic tropical sugarbeet
Aranka, Cauvery, SZ 35, PAC 60008, Danicia, Sereneda, Natura performed better in
respect of beet yield, yield contributing characters and sugar recovery. Hence, these
genotypes can be successfully cultivated in Bangladesh.
Performance of Some Exotic Tropical Sugarbeet ......... Bangladesh 9
REFERENCES
Aly, E.F. 2006. Effect of environmental conditions on productivity and quality of some
sugarbeet varieties. Ph. D. Thesis. Fac. of Agric. Benha Univ. Egypt.
Amr, A.H.R. and Ghaffar M.S.A. 2010. The economic impact of sugarbeet cultivation in
new lands (Study of Al-Salam Canal Area Status). Aust. J. Basic & Appl. Sci., 4
(7): 1641-1649.
Anonymous. 2005. Tropical sugarbeet production technology, soil and crop
management studies, Tamil nadu Agricultural U niversity, Coimbatore, India. p.
10.
Asadi, M. 2007. Sugarbeet processing in Beet-Sugar Handbook. John Wiley & Sons Inc.,
New Jersey, USA. pp. 100-465.
El-Bakary, H.M.Y. 2006. Studies on yield and quality characters of some sugarbeet
varieties. M.Sc. Thesis Fac. of Agric Al-Azhar Univ. Egypt.
Follet, R.H.; Schmehl, E.R.; Powers L. and Payne, M.G. 1964. Effect of genetic
population and soil fertility level on the chemical composition of sugarbeet tops.
Tech. Bull. Colombia Agric. Expt.
Ismail, A.M.A.; Al-Labbody, A.H.S. and Shalaby, N.M.S. 2006. Variability and traits
relationship in nine sugarbeet varieties under three sowing dates. Egypt. J. Plant
Breed., 10 (1): 387-406.
Kapur, M.L. and Kanwar R.S. 1990. Phosphorus fertilization in subtrop ical India. J.
Sugarbeet.
Memon, Y.M.; Khan I. and Panhwar R.N. 2004. Adoptability performance of some exotic
sugarbeet varieties under agro-climatic conditions of Thatta. Pakistan Sugar J.,
19 (6): 42-46.
Osman, A.M.H.; El-Sayed G.S.; Osman, M.S.H. and El-Sogheir K.S. 2003. Soil
application of some microelements with relation to yield and quality of sugarbeet
varieties. Annals of Agric. Sc., Moshtohor, 41 (3): 1071-1088.
Petkeviciene, B. 2009. The effects of climate factors on sugarbeet early sowing timing.
Agron. Res., 7: 436–443.
Rahman, M.K.; Kabir, M.L.; Alam, M.J.; Hossain, M.S. and Islam, A.K.M.R. 2006.
Sugarbeet cultivation in Bangladesh. Bangladesh Sugarcane Research Institute
Ishurdi, Pabna, Bangladesh.
Urich, A. 1952. The influence of temperature and light factors on the growth and
development of sugarbeets in controlled conditions. Agron. J., 44: 66-73.
Bangladesh J. Sugarcane, 35 : 10-19 June, 2014
Effect of Nitrogen and Potassium on Growth, Development
and Sugar Accumulation in Tropical Sugarbeet
M.N. Kashem1*, Q.A. Khaliq2, M.A. Karim2, A.J.M.S. Karim2, S.M.R. Karim1 and
M.M. Hossain1
2 Professor, Agronomy Department, BSMRAU, Gazipur
1 Training and Technology Transfer Division
Bangladesh Sugarcrop Research Institute, Ishurdi-6620, Pabna, Bangladesh
ABSTRACT
A field experiment was conducted at the Agronomy R esearch Farm,
Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur,
Bangladesh from November 2012 to April 2013 with four levels of nitrogen viz.,
0, 50, 100 and 150 kg N ha-1 in combination with four levels of potassium viz.,
0, 60, 120 and 180 kg K ha-1 to findout the optimum doses of nitrogen and
potassium for optimum growth, development and sugar accumulation in
tropical sugarbeet in Bangladesh. Plant height, number of leaves per plant, LAI
and crop growth rate (CGR) significantly increased with the increase in
nitrogen level but not significant with increased potassium level. The
combination of 150 kg N and 180 kg K ha-1 resulted in the highest number of
leaves per plant and LAI. Increasing nitrogen levels led to decrease in TSS
content in juice and sucrose content in root. The TSS content increased with
increasing potassium level up to 120 kg K ha-1. The combination of nitrogen
and potassium levels did not show any significant effect on sucrose content in
root. Increasing nitrogen and potassium levels caused reduction in juice purity.
For optimum growth, development and sugar accumulation in tropical
sugarbeet in Bangladesh the nitrogen and potassium requirement of the crop
is 150 kg N and 180 kg K ha-1.
Key words: Nitrogen, potassium, growth, development, sugar
accumulation, tropical sugarbeet
INTRODUCTION
Nitrogen is one of the most important nutrient elements of those supplied to
sugarbeet in fertilization. It causes desirable effect on sugarbeet growth and
developments. In New Zealand, sugar yields were optimal when nitrogen was applied at
the rate of 200 kg N ha-1, (Smit et al., 1995). The increase in root and sugar yield with N
fertilizer is attributed to increased size and number of leaves, which lead to increased leaf
area and photosynthetic activities. An adequate supply of N is essential for optimum yield
but excess of it may result in an increase in yield of roots with lower sucrose content and
juice purity. High rates of N fertilizer (214 kg and 285 kg N ha-1) significantly increase the Na
and α- amino-N content in root juice (Abdel-Motagally et al., 2009). The high rate of N
increases soluble non-sugar impurities in root juice and the percentage of sugar losses to
molasses and consequently reduces sugar recovery. El-Shahawy et al. (2002) and Attia,
* Corresponding author: M. N. Kashem, Principal Scientific Officer
e-mail: [email protected]
Effect of Nitrogen and Potassium on Growth ........... Sugar Beet 11
(2004) found significant decrease in Fe and Mn content in foliage and roots with high rate
of N fertilization. Sugar beet is a high potassium (K) requiring crop. Ibrahim et al. (1998)
found the highest sucrose percentage and juice purity with K application up to 228.5 kg
K2O ha-1. The beneficial effect of K fertilization on growth, yield and quality of sugarbeet
was emphasized by previous studies (Sobh et al., 1992; El-Shafai, 2000). It is commonly
observed that root enlargement is depressed more than leaf development, when K is in
short supply. El-Shafai (2000) and O'shea et al. (2009) found that nitrogen and potassium
fertilization significantly influences the sugarbeet quality. Hence, the present research
work was undertaken to findout the optimum doses of nitrogen and potassium fertilizer for
growth, development and sugar accumulation in tropical sugar beet under Bangladesh
condition.
MATERIALS AND METHODS
The experimental site was belongs to Madhupur Tract (AEZ 28) that
characterized by a sub-tropical climate. The experiment was carried out from November
2012 to April 2013. The experimental land was prepared thoroughly by plouphing. As
sugar beet prefers alkaline soil dolomite was applied @ 1500 kg ha-1 (Islam et al., 2010).
During final land preparation cow dung @ 15 t ha-1 was incorporated into the soil. A
fertilizer dose of 120 kg N, 105 kg P, 150 kg K, 18 kg S, 3.5 kg Zn and 1.2 kg B ha-1 was
applied in the form of urea, TSP, MOP, Gypsum, ZnSO4 and Boric acid, respectively. All
fertilizers and 1/3 of urea were applied during final land preparation. The remaining
amount of urea was applied as two top dressing 55 and 90 DAS. The experiment was laid
out in a strip plot with three replications. The unit plot size was 3m × 2m. The experiment
comprised of following treatments:
Factor A: Four levels of nitrogen viz., 0, 50, 100 and 150 kg N ha-1
Factor B: Four levels of potassium viz., 0, 60, 120 and 180 kg K ha-1
Seeds of tropical sugar beet genotype (Shubhra) were sown in lines on 01 November,
2012 with the spacing of 50 x 20 cm. Light irrigation was done immediately after sowing
to ensure uniform emergence. To ensure optimum soil moisture irrigation was done twice
in a week up to maturity till April. Intercultural operations were done uniformly in each plot
to ensure normal growth of the crop. Weeding and mulching were done simultaneously in
the experimental plots at 20, 40 and 60 DAS. Plant was thinned out keeping one plant per
hill during the second weeding. Earthing up was done at 55 and 90 DAS after top
dressing of nitrogen. Dithane M 45 @ 2.2 kg ha-1, Tilt 1ml/L of water and Score 250 EC
0.5 ml/L of water were used to control damping off, sclerotium root rot and cercospora
leaf spot diseases. Durshban @ 2.5 ml/L of water was applied for controlling cut warm,
tobacco caterpillar and army warm. Data regarding number of leaves, leaf area index,
crop growth rate, relative growth rate, net assimilation rate, total soluble solid, sucrose
content and sucrose yield were collected, analyzed and interpreted. Beets were
harvested calculated (kg m-2 and t ha-1) and statistically analyzed with the help of MSTAT-
C Program with LSD Test at 5% level of significance.
Following formulae were used to calculate different growth parameters
LAI = Total leaf area of three plants (cm2)
Ground area covered by three plants (cm2)
12 Bangladesh J. Sugarcane, 35 : 10-19 June, 2014
Crop Growth Rate W2 – W1 g m-2 day-1
1 ----------------
T2 – T1
CGR = ------- ×
GA
Relative Growth Rate
(Ln W2 – LnW1)
RGR = ----------------------------- (mg g-1day-1)
T2 – T1
Where,
W1 = dry weight at time oTr1L(agn),dWa2re=ad(rmy2w) eight at time T2 (g) and
GA = ground area (m2)
RESULTS AND DISCUSSION
Plant height, number of leaves per plant and leaf area index (LAI)
It was found that the plant height, number of leaves per plant and LAI increased
rapidly up to 120 DAE and thereafter followed a slower rate. The general trend was an
increase in plant height, number of leaves per plant and LAI with the increase in nitrogen
level. Plant height is an important morphological character that might affect yield.
Nitrogen is responsible for rapid growth of a plant. The height of a plant depends on
nutrient availability especially on nitrogen. The variations in plant height over time are
illustrated in Figure1 reveals that different levels of nitrogen fertilizer exerted significant
influence on plant height throughout the growth periods. Irrespective of treatments plant
height increased rapidly up to 120 DAE and thereafter a slower rate was found. During
the growth periods at 120 DAE the tallest plant (41.67 cm) was obtained at 150 kg N ha-1
and the shortest (29.08 cm) was at 0 kg N ha-1.
Combination effects of nitrogen and potassium levels on number of leaves per
plant in tropical sugar beet started functioning significantly from 90 DAE (Table1). The
highest number of leaves (36.00 plant-1) was recorded in combination of 150 kg N ha-1
and 180 kg K ha-1 at 150 DAE followed by the 150 kg N ha-1 and 0-120 kg K ha-1, 150 kg
N ha-1 and 0-120 kg K ha-1 and 150 kg N ha-1 and 100 kg K ha-1at same DAE which were
statistically identical, while the lowest number of leaves was recorded at 0 kg N ha-1 with
0 kg K ha-1 (31.67 plant-1). It was revealed that 150 DAE was the turning point of leaf
induction because older leaves dropped. However, Watson et al. (1972) found that the
number of leaves in sugarbeet is relatively unaffected by cultural practices or
environmental factors. At 165 DAE, the leaves counted were less than those at 150 DAE,
and the highest number of leaves retained in the treatment combination of 100-150 kg N
ha-1 with 120-180 kg K ha-1. Therefore, after assessing the data it may be concluded that
sugarbeet genotype shubhra performs the best with the treatment combination of 100-
150 kg N ha-1 × 120 kg K ha-1 to have the highest number of leaves per plant.
Leaf area index of tropical sugarbeet genotype shubhra over time was
significantly influenced by different levels of nitrogen (Figure 2). Irrespective of nitrogen
levels, the LAI increased sharply after emergence up to 90 DAE then gradually attained a
Effect of Nitrogen and Potassium on Growth ........... Sugar Beet 13
peak at 120 DAE and thereafter decreased gradually. This increase was due to the
production of higher number of functioning leaves, which increased total photosynthetic
area. Similar trend was also reported by Theurer (1979) in sugarbeet. Different nitrogen
levels strongly influenced the LAI at 60, 90, 120, 150 and 165 DAE. At 120 DAE, the
highest LAI (3.47) was obtained with 150 kg N ha-1 followed by 100 kg N ha-1 (2.75) and
50 kg N ha-1 (1.90) and the lowest LAI (1.53) was found when no nitrogen was applied.
Figure 1. Plant height in tropical sugar beet genotype shubhra over time as influenced by
different levels of nitrogen application. Vertical bar indicates LSD 0.05.
Table 1. Number of leaves per plant in tropical sugarbeet genotype shubhra over
time
Treatment Number of leaves per plant
(N×K)
30 DAE 60 DAE 90 DAE 120 DAE 150 DAE 165 DAE
N0 K0 6.33 29.67
N0 K60 6.33 13.00 19.33 30.33 31.67 30.33
N0 K120 6.33 30.00
N0 K180 6.33 13.67 19.33 31.00 32.00 30.00
N50 K0 6.66 31.67
N50 K60 6.33 14.33 19.67 31.67 32.00 31.67
N50 K120 6.33 32.00
N50 K180 14.00 19.33 31.00 32.00
N100 K0 6.66 32.67
N100 K60 6.33 14.33 20.67 32.00 33.33 32.00
6.33 32.33
14.33 20.67 32.00 33.67
14.33 21.00 32.67 34.00
14.00 21.00 32.67 34.33
14.33 21.33 33.33 34.67
14.67 21.67 33.00 35.00
(Contd.)
14 Bangladesh J. Sugarcane, 35 : 10-19 June, 2014
Treatment Number of leaves per plant
(N×K)
30 DAE 60 DAE 90 DAE 120 DAE 150 DAE 165 DAE
N100 K120 6.66 32.67
N100 K180 6.66 15.00 21.67 33.67 35.33 33.67
N150 K0 6.33 32.67
N150 K60 6.33 15.33 22.00 33.67 35.33 33.00
N150 K120 6.66 33.67
N150 K180 6.66 14.33 22.00 33.67 35.67 33.67
LSD (0.05) NS 1.75
CV (%) 9.00 15.00 22.33 33.33 35.67 3.29
15.00 22.33 33.67 35.67
15.33 22.67 33.67 36.00
NS 1.37 2.04 2.51
7.10 3.91 3.75 4.41
Figure 2. Leaf area index in tropical sugar beet genotype shubhra over time as influenced
by different levels of nitrogen application. Vertical bar indicates LSD 0.05.
Crop Growth Rate
Nitrogen and potassium exerted significant effects on crop growth rate. The
tropical sugarbeet showed the highest CGR (24.91 g m-2day-1) with the combination
levels of 150 kg N ha-1 and 180 kg K ha-1 followed by (24.83 g m-2day-1) 150 kg N ha-1
and 120 kg K ha-1 and 100 kg N ha-1 and 180 kg K ha-1 which are statistically identical.
Lower rate of fertilizer combination resulted in lower crop growth rate throughout the
growth period which ultimately lowered the yield of tropical sugarbeet. The CGR
gradually increased with each increase in nitrogen and potassium combination levels
over time. The increase in CGR due to raising nitrogen and potassium fertilizer levels
may be attributed to their role in increasing LAI and dry matter accumulation in root and
foliage. So that 100 - 150 kg N ha-1 and 0 -180 kg K ha-1 will be the suitable combination
for sugarbeet cultivation.
Effect of Nitrogen and Potassium on Growth ........... Sugar Beet 15
Table 2. Combination effect of nitrogen and potassium levels on crop growth rate
in tropical sugarbeet genotype shubhra over time
Treatment Crop growth rate (g m-2 day-1)
(N×K)
30 -60 DAE 60 -90 DAE 90-120 DAE 120-150 DAE 150-165 DAE
N0 K0 3.26 2.71
N0 K60 3.31 8.28 12.64 10.99 2.73
N0 K120 3.50 2.65
N0 K180 3.47 8.48 12.29 11.84 3.18
N50 K0 4.99 3.38
N50 K60 5.07 8.75 13.61 11.95 3.59
N50 K120 5.46 3.74
N50 K180 5.52 8.68 13.81 12.23 3.13
N100 K0 6.29 3.02
N100 K60 6.41 9.56 17.43 21.11 3.22
N100 K120 6.57 4.15
N100 K180 6.66 9.63 17.57 21.80 4.38
N150 K0 6.97 3.74
N150 K60 7.08 9.86 18.28 22.25 3.94
N150 K120 7.14 4.36
N150 K180 7.16 9.93 18.48 22.29 4.82
LSD (0.05) 1.38 1.43
CV (%) 14.94 11.11 22.09 22.84 14.19
11.44 22.45 23.29
11.49 22.84 23.77
11.50 22.89 23.99
13.54 24.29 24.32
13.94 24.33 24.36
14.93 24.83 24.38
15.14 24.91 24.87
2.24 3.23 2.94
12.22 9.90 8.64
Relative Growth Rate (RGR)
Relative growth rate of tropical sugarbeet over time did not significantly differ
either by nitrogen levels nor by potassium levels. Irrespective of nitrogen levels, the RGR
was high in early growth period and showed similar decreasing trend as the crop
advanced in age. The decline in RGR towards maturity might be due to lower
photosynthetic efficiency of older leaves as well as leaf senescence. The decline in RGR
was also due to the decrease in net assimilation rate. Rao and Mitra (1988) and Singh et
al. (1998) also reported the similar results in groundnut and in mungbean, respectively.
16 Bangladesh J. Sugarcane, 35 : 10-19 June, 2014
Figure 3. Relative growth rate (mg g-1day-1) in tropical sugarbeet genotype shubhra over
time as influenced by nitrogen levels.
Table 3. Combination effect of nitrogen and potassium levels on TSS %,
sucrose%, apparent purity % and sugar yield in tropical sugarbeet
genotype shubhra at maturity
Treatment TSS % Sucrose % Apparent Sugar Yield
(N×K) purity % (t ha-1)
23.12 15.57 6.90
N0 K0 22.94 15.60 67.23 7.63
N0 K60 23.15 15.68 67.61 7.52
N0 K120 23.27 15.76 67.57 7.52
N0 K180 23.02 15.42 67.46 8.53
N50 K0 23.08 15.45 66.76 9.47
N50 K60 22.75 15.40 66.92 9.45
N50 K120 22.67 15.30 67.42 9.44
N50 K180 22.30 15.03 67.53 10.58
N100 K0 22.31 15.18 67.31 11.14
N100 K60 22.35 15.20 67.32 12.49
N100 K120 22.44 15.21 67.42 12.55
N100 K180 22.10 14.83 67.42 11.43
N150 K0 66.92
22.12 14.95 12.17
N150 K60 22.33 15.00 66.76 12.74
N150 K120 22.23 14.98 66.14 13.07
N150 K180 0.52 66.39 1.60
LSD (0.05) ns 3.07 9.44
CV (%) 2.99 ns
1.62
Effect of Nitrogen and Potassium on Growth ........... Sugar Beet 17
Total soluble solids (TSS)
Nitrogen and potassium levels did not exert significantly differed on TSS at
maturity Table 3. The results showed that the higher percentages of total soluble solids
were obtained at lower levels of nitrogen and potassium combinations. The TSS was
reduced with highest nitrogen and potassium combination levels. The decrease in TSS
content due to excessive nitrogen or potassium application can be ascribed to their role in
increasing root weight, root diameter, tissue water content. This result confirms the
findings of Sobh et al. (1992) and Sultan et al. (1999).
Sucrose content
Nitrogen and potassium combination levels significantly influenced the sucrose
content in roots of tropical beet. It was observed that each increase in nitrogen and
potassium combination levels from 0 kg N ha-1 and 0 kg K ha-1 to 150 kg N ha-1 and 180
kg K ha-1 was associated with gradual decrease in sucrose content. The maximum
sucrose content in beet roots was recorded from low levels of nitrogen and potassium
combination. On the contrary, the lowest one was recorded from application of 150 kg N
ha-1 and 180 kg K ha-1. The decrease in sucrose content owing to increasing nitrogen
fertilizer levels can be attributed to its role in increasing non-sucrose substances such as
proteins and alpha amino acid, and hence decreasing sucrose content in roots. Similar
results were obtained by Sultan et al. (1999).
Sugar yield
Sucrose yield is the most important parameter in sugarbeet. Nitrogen and
potassium levels had significant effect on sugar yield in tropical sugar beet. Remarkable
increase in sugar yield was noticed as a result of increased nitrogen and potassium
combination levels from 0 kg N and 0 kg K ha-1to 150 kg N and 180 kg K ha-1. The
nitrogen and potassium combination, which produced the highest sugar yield (13.07 t ha-1)
was 150 kg N and 180 kg K ha-1 followed by 150 kg N and 120 kg K ha-1 (12.74 t ha-1),
100 kg N and 180 kg K ha-1 (12.55 t ha-1) and 100 kg N and 120 kg K ha-1 (12.49 t ha-1).
However, excess nitrogen application might not be desirable, because it reduces the
most quality parameters and sugar yield. The increase in gross sugar yield per unit area
due to application of nitrogen and potassium fertilizers can be explained through the fact
that nitrogen and potassium play a vital role in improving all growth attributes and root
weight per plant as well as sucrose content in root, consequently increasing gross sugar
yield per unit area. These results agree with those stated by EL-Hawary (1999), Sultan et
al. (1999) and EL-Zayat (2000).
Apparent purity percentage
Apparent purity percentage of tropical sugarbeet was influenced by nitrogen,
potassium and their combination levels. Increasing nitrogen fertilizer levels from 0 to 50,
100 and 150 kg N ha-1 tended to decrease juice purity from 67.18 to 67.15, 67.15 and
67.08 %, respectively. Similar results were obtained by Abd EL-Moneim (2000), Kandil et
al. (2002b) and Ramadan et al. (2003). Increasing potassium fertilizer levels from 0 to 60,
120 and 180 kg K ha-1 tended to increase juice purity from 66.58 to 67.49, 67.78 and
67.98%, respectively. Similar trend was obtained by Kandil et al. (2002a). Increasing
nitrogen and potassium combination levels from 0 kg N ha-1 and 0 kg K ha-1 to 150 kg N
ha-1 and 180 kg K ha-1 caused reduction in juice purity in all cases. The reduction in juice
18 Bangladesh J. Sugarcane, 35 : 10-19 June, 2014
purity owing to increased nitrogen fertilizer levels can be attributed to its role in increasing
impurity substances such as proteins and alpha amino acid, and hence decreasing juice
purity. Similar results were reported by Abou-Amou et al. (1996).
From the experimental results in the field, it may be concluded that for optimum growth,
development and sugar accumulation in tropical sugarbeet in Bangladesh. The nitrogen
and potassium requirement of the crop seems to more than 150kg N and 180kg K ha-1.
Hence, excess nitrogen application not desirable, because it reduces the most quality
parameters and sugar yield. The increase in gross sugar yield per unit area due to
application of nitrogen and potassium play a vital role in improving all growth attributes
and root weight per plant as well as sucrose content in root, consequently increasing
gross sugar yield per unit area.
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and quality of some sugarbeet varieties grown in Fayoum region. Ph. D. Thesis,
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to levels and times of potassium fertilization under salinity conditions at northern
Delta. Egypt. J. Agri. Sci., 27(11): 7237-7246.
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O'shea, C.J.; Lynch, B.; Lynch, M.B.; Callan J.J. and O'Doherty, J.V. 2009. Ammonia
emissions and dry matter of separated pig manure fractions as affected by crude
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Bangladesh J. Sugarcane, 35 : 20-27 June, 2014
Germination and Yield of Tropical Sugarbeet as Influenced
by Variety and Fertilizer
S. Islam1*, M.A.T. Sohel2, M.A. Haque1, M.A. Razzak2, M.J. Islam3, S.M.R. Karim4,
M.S. Islam1 and K.M. Alam1
1 Soils and Nutrition Division, 2Agronomy and Farming Systems Division
3 Physiology and Sugar Chemistry Division, 4Training and Technology Transfer Division
Bangladesh Sugarcrop Research Institute, Ishurdi-6620, Pabna, Bangladesh
ABSTRACT
An experiment was conducted at Bangladesh Sugarcrop
Research Institute farm, Ishurdi during 2013-14 cropping season to
investigate the effects of varieties and fertilizer levels on seed germination and
yield of tropical sugarbeet. Significant differences were observed in percent
germination, yield and brix% of beet. It was afnodunbdeethtayt ievladrie(6ty0.8V2th(Ca-a1)uvoevreyr)
produced the highest germination (72.75%)
the variety V1 (Shubrha) (with 58.53 % and 57.6 tha-1) respectively. In case of
fertilizer, the treatment T3 (MOC625 N150 P25 K140 S22 Zn4 Bth1a.5-1k)gamhao-1n)gparolldoutcheedr
the highest germination (72.79%) and beet yield (101.0
treatments. Results from the interaction of variety and fertilizer revealed that,
vkagrhieaty-1)Vt2re(aCtamuevnetryo)bwtaiitnhetdhethfeerhtiilgizheerslet vbeeleTt 3yi(eMldO(C160245.N67150thPa2-51)Kw14h0eSre22 Zn4 B1.5
in every
cases Control plot obtained (thMeOClo6w25eNst150yPie2ld5 .K1T40hSer2e2 fZonre4,B1t.h5ekgvhaari-e1)tymaVy2
(Cauvery) and fertilizer level T3
be recommended for cultivation of tropical sugarbeet at Ishurdi under AEZ -11
of Bangladesh.
Key words: Sugarbeet, germination, fertilizer dose, brix %, beet yield
INTRODUCTION
Sugarbeet (Beta vulgaris L.) ranks the second after sugarcane in terms of world’s
sugar production. Sugarbeet is a temperate crop, generally grown in Europe, North
America and temperate zones of Asia. France, Germany, Russia, USA and Ukraine are
the most sugarbeet producing countries of the world (FAO, 2010). Although sugarbeet is
a temperate crop, the international company Syngenta developed and successfully
introduced a new sugarbeet that can be grown under tropical climatic conditions and
hence, the beet is known as tropical sugarbeet. Substantial yield increase of sugarbeet is
essential which may vary from variety to variety (Abo-Salama and EL-Sayiad, 2000). So it
is crucial to find out a specific variety to obtain maximum beet yield from tropical
sugarbeet in Bangladesh. Fertilizer was considered as a limiting factor for obtaining
higher yield and superior quality in sugarbeet (Ouda, 2002). Thus, application of nitrogen,
phosphorus and potassium fertilizers is immense importance for the production of
sugarbeet. O'shea et al. (2009) reported that increased the yield and quality of sugarbeet
mainly depended on fertilizer, especially in N and K. The increase in beet yield with N
fertilizer may be attributed to increased size and number of leaves, which led to increased
*Corresponding author: S. Islam, Scientific Officer
e-mail: [email protected]
Germination and Yield of Tropical Sugarbeet as .......... Fertilizer 21
leaf area and photosynthetic activities. An adequate supply of N is essential for optimum
yield, but excess of it may result in an increase in yield of roots with lower sucrose
content and juice purity. Sugarbeet was found as high potassium (K) requiring crop
(Johanson et al., 1971). The role of K fertilization on growth, yield and quality of
sugarbeet was emphasized by previous studies conducted by El-Maghraby et al. (1998)
and El-Shafai (2000). There is a scanty of research in developing a fertilizer
recommendation for sugarbeet production in Bangladesh. Considering the above facts,
the present study was undertaken to evaluate the suitable variety and estimate optimum
doses of fertilization for maximum yield of tropical sugarbeet at Ishurdi in Bangladesh.
MATERIALS AND METHODS
The experiment was conducted at BSRI farm, Ishurdi under High Ganges River
Flood Plain Soils (AEZ 11) during 2013-14 cropping season. The experiment was laid out
randomized complete block in factorial design with three replications. There were four
fTk(eNg3r-)ht1ialM2iz0-1Oe,PrCihnl6oe2scv5poeNhmls1o5br0vuiniPsza.2,t(5iPoTK)n112-400wCPSitooh2nt2atZtrswonsol4iu(BmNs1uo.5g(Kafke)rg1rbht1ei2lai.e5z-1teSravu)na,lpdrTihe2Tu-ti4re-M(sMSuv)sO1itz8aC.Zr,3di7nV5Oc1Ni(9-lZ0CSnPa)h13k5u.5ebKBr8(ho4Ma.r3oO8, nSCV12()3B5.05-)01ZC.2Nna2iktu.6rgvo3hegBare0-y1n.9.,
Sugarbeet seeds were sown on 05 November, 2013 at 50cm × 20cm spacing in 5m × 4m
sized plots. Grined MOC was incorporated in the plots of the respective treatments at 6
days before seeds sowing. Full amount of PKSZnB and one third of N were applied in soil
as basal and thoroughly mixed with soil. The rest amount of N was applied in two equal
splits at 30 and 60 DAS. The cultural operations such as irrigation, weeding, earthing up
and pest control were done as and when necessary. The crop was harvested on 03 May,
2014. Data on seed germination, beet yield and brix % were collected and analyzed by
statistix-10 software.
RESULTS AND DISCUSSION
Germination percent of tropical sugarbeet
Effect of variety
Figure 1 represents percent germination of sugarbeet varieties. The highest
germination percent was found in Cauvery (72.75%) over V1 Shubhra (58.53%). This
result was supported by Arshadi and Asgharipour (2011) where they reported that there
had a significant difference between the germination percent in different genotypes of
sugarbeet.
22 Bangladesh J. Sugarcane, 35 : 20-27 June, 2014
Figure 1. Effect of variety on seed germination of tropical sugarbeet varieties
Effect of fertilizer
Results revealed that the germination percentages were significantly influenced
by different fertilization levels Table 1. The treatment T3 (MOC625 N150 P25 K140 S22 Zn4 B1.5
kgha-1) produced the highest germination (72.79%), but the effect was statistically similar
with T2 (MOC500 N120 P20 K112.5 S18 Zn3.5 B1.2 kgha-1), and the lowest percentage was
obtained by the T1 (Control) treatment (56.83%). Stevens et al. (2011) reported that
fertilizer in band placement between 7.5 and 12.5 cm deep (5 to 10 cm below the seed)
resulted with the best combination of N uptake and seedling emergence.
Interaction effect of variety and fertilizer
Interaction effect of variety and fertilizer had significantly influenced on
germination percentages Table 1. The highest germination (80.92%) was found from the
variety V2 (Cauvery) with fertilizer level T3 (MOC625 N150 P25 K140 S22 Zn4 B1.5 kgha-1), which
was statistically similar to the same variety with T2 (MOC500 N120 P20 K112.5 S18 Zn3.5 B1.2
kgha-1) treatment, and the lowest germination (51.08%) was produced by the variety V1
(Shubhra) with T1 (Control) treatment.
Germination and Yield of Tropical Sugarbeet as .......... Fertilizer 23
Table 1. Seed germination influenced by fertilizer and interaction effect of variety
and fertilizer
Treatment Germination %
Fertilizer levels:
56.83 c
T1 68.75 ab
T2 72.79 a
T3 64.18 b
T4
LSD (0.05) 5.07
Fertilizer ×Variety: 51.08 d
T1V1 59.92 c
T2V1 64.67 bc
T3V1 58.43 c
T4V1 62.58 c
T1V2 77.58 a
T2V2 80.92 a
T3V2 69.92 b
T4V2
7.18
LSD (0.05)
T1- Control (No fertilizer), T2- MOC500 N120 P20 K112.5 S18 Zn3.5 B1.2 kg ha ,
T3- MOC625 N150 P25 K140 S22 Zn4 B1.5 kg ha-1, T4- MOC375 N90 P15 K84.38 S13.5 Zn2.63
B0.9 kg ha-1, V1 - Shubrha, V2 - Cauvery
Beet yield
Effect of variety
Data on beet yield showed that, there were significant differences among the
varieties Table 2. The highest beet yield (60.80 t ha-1) was obtained in Cauvery and the
lower yield (57.6 t ha-1) produced in Shubrha. Nenadić et al. (2003) reported that the
Swedish cultivar Dorotea (tolerant to both rhizoctonia and cercospora) was found as the
highest yielding crop. The lowest yielding crop (susceptible to both rhizoctionia and
cercospora) was the domestic cultivar Dana. However, planting sugarbeet V2 (Cauvery)
was observed more favorable for higher sugar yield at the study location at Ishurdi in
Bangladesh.
Effect of Fertilizer
Sugarbeet yield was significantly influenced by fertilizer levels was observed in
Table 2. The highest beet yield (101.0 t ha-1) was found from the treatment T3 (MOC625
N150 P25 K140 S22 Zn4 B1.5 kgha-1) and the lowest was obtained from T1: treatment (3.1 t ha-1).
Kashem (2014) stated that increasing nitrogen levels from 0 to 50 kg or from 50 to100
kg N ha-1 caused increase in beet yield by 26.80 and 29.03%, respectively. Similar
findings were reported by Abd EL-Moneim (2000); EL-Shahawy et al. (2002) and
Ramadan et al. (2003) in their study. In New Zealand, sugarbeet root and sugar yields
24 Bangladesh J. Sugarcane, 35 : 20-27 June, 2014
were optimal when nitrogen was applied @ 240 and 200 kg ha-1, respectively (Smit et al.,
1995). They further reported that fresh top weight increased with increasing nitrogen
availability, while sugar content was reduced.
Table 2. Effect of variety and fertilizers on yield and brix% of tropical sugarbeet
Treatment Yield (t ha-1) Brix (%)
Variety:
57.6 b 17.15 b
V1 60.8 a 18.35 a
V2 2.0257
LSD (0.05) 0.30
Fertilizer levels: 3.1d
T1 72.6 b 18.57 a
T2 101.0 a 17.53 b
T3 60.5 c 16.97 c
T4 2.8648 17.92 b
LSD (0.05)
0.42
T1- Control (No fertilizer), T2- MOC500 N120 P20 K112.5 S18 Zn3.5 B1.2 kg ha ,
T3- MOC625 N150 P25 K140 S22 Zn4 B1.5 kg ha-1, T4- MOC375 N90 P15 K84.38 S13.5 Zn2.63
B0.9 kg ha-1, V1 - Shubrha, V2 - Cauvery
Interaction effect of variety and fertilizer
Beet yield was significantly influenced by interaction between variety and fertilizer
Table 3. The highest beet yield (104.67 t ha-1) was produced in interaction between the
variety Cauvery tanhda-1M) OwCa6s25rNec15o0rdPe25dKw14i0thS2t2hZeni4ntBe1r.a5 cktgiohna-1bfeetrwtielizeenr dose, and the lowest
beet yield (2.9 Cauvery variety and
Control (No fertilizer) treatment. These results indicated that sugar beet variety Cauvery
should be grown with the fertilizer dose MOC625 N150 P25 K140 S22 Zn4 B1.5 kgha-1 in High
Ganges River Flood Plain Soils (AEZ 11) for achieving higher sugar yield.
Brix % of beet
Effect of variety
Significant differences were recorded for brix % among the varieties Table 2. The
highest brix % (18.35%) was recorded from V2 (Cauvery) and the lowest (17.15) from the
V1 (Shubrha) treatment.
Effect of Fertilizer
Fertilizer levels significantly influenced brix % in sugarbeet Table 2. The highest
brix % (18.57 %) was found from the treatment T1: Control plot and the lowest (16.97 %)
was obtained from the treatment T3 (MOC625 N150 P25 K140 S22 Zn4 Bh1a.5-1kglehda-t1o).dKeacrsehaesme
(2014) reported that increasing nitrogen levels from 100 to 150 kg N
in brix % content by 2.25 %. Similar findings were reported by Attia et al. (1999);
Mahasen (1999) and Nemeat Alla et al. (2002).
Germination and Yield of Tropical Sugarbeet as .......... Fertilizer 25
Table 3. Interaction effect of variety and fertilizers on yield and brix % of tropical
sugarbeet
Variety × Fertilizer Yield (t ha-1) Brix (%)
T1V1 3.23 f 17.95 c
T2V1 69.50 d 17.00 d
T3V1 97.33 b 16.36 e
T4V1 60.13 e 17.29 d
T1V2
T2V2 2.90 f 19.19 a
T3V2 76.00 c 18.06 bc
T4V2 104.67 a 17.58 cd
61.20 e 18.56 b
LSD (0.05) 4.0514
0.59
T1- Control (No fertilizer), T2- MOC500 N120 P20 K112.5 S18 Zn3.5 B1.2 kg ha-1,
T3- MOC625 N150 P25 K140 S22 Zn4 B1.5 kg ha-1, T4- MOC375 N90 P15 K84.38 S13.5 Zn2.63
B0.9 kg ha-1, V1 - Shubrha , V2 - Cauvery
Interaction effect of variety and fertilizer
Brix percentage was significantly influenced by interaction between variety and
fertilizer Table 3. The highest brix (19.19 %) in sugarbeet was produced by interaction
between Cauvery variety and Control (No fertilizer) fertilizer dose, and the lowest brix %
(16.36 %) of sugarbeet was recorded with the interaction between Shubrha variety and T3
(MOC625 N150 P25 K140 S22 Zn4 B1.5kgha-1) fertilizer dose.
Economic performance
The economic performance of different levels of variety and fertilizer doses on
sugarbeet (AEZ 11) is presented in Table 4. The highest Marginal Benefit Cost Ratio
(SNM2125B0ZCPnR425)BK611..5420k7Sghw22aaZ-s1nf4eorbBtti1lai.z5inekergdihnainc-1uiflnetitrvetairlitaziocentriodonof ssbeue.gtwaHreebenenceett,hveaaprvipealtirycieaCttyiaounCvaoeufryvMeaOrtyCIsa6h2n5udNrdT1i520(AMPE2OZ5 CK11614205)
was found the most economically profitable.
Table 4. Economic performance of sugarbeet varieties at different fertilizers level
at BSRI, Ishurdi.
Treatment Fertilizer Yield Gross Net Benefit MBCR (1)
T1V1 cost (tha-1) return
T2V1 (Tk.ha-1) -
T3V1 (Tk. ha-1) 3.23 5.03
T4V1 69.50 9,690 9,690 5.76
T1V2 0 97.33 5.96
T2V2 34,548 60.13 2,08,500 1,73,952
T3V2 43,175 -
T4V2 25,911 2.90 2,91,990 2,48,815 5.59
76.00 6.27
0 104.67 1,80,390 1,54,479 6.08
34,548 61.20
43,175 8,700 8,700
25,911
2,28,000 1,93,452
3,14,010 2,70,835
1,83,600 1,57,689
26 Bangladesh J. Sugarcane, 35 : 20-27 June, 2014
Input Price: Urea= Tk. 16.00 kg-1. TSP= Tk. 22.00 kg-1, MOP= Tk. 15.00 kg-1, Gypsum= Tk. 15.00
kg-1, Zinc Sulphate Mono: Tk. 120 kg-1, Boric Acid= 240 kg-1, Mustard Oil Cake: Tk. 40
kg-1.
Variable cost includes fertilizer cost only
Output Price: Sugarbeet: Tk. 3000.00 t-1
From the present findings, it may be concluded that the variety Cauvery and the fertilizer
rate of MOC625 N150 P25 K140 S22 Zn4 B1.5 kgha-1 were found appropriate for achieving
higher yield of tropical sugarbeet at the study location of BSRI, Ishurdi under High
Ganges River Flood Plain Soils (AEZ 11).
REFERENCES
Abd EL-Moneim, S.A. 2000. Effect of preceding crop and nitrogen fertilizer levels on yield
and quality of some sugarbeet varieties grown in Fayoum region. Ph. D. Thesis,
Fac. of Agric., EL-Fayoum, Cairo Univ.
Abo-Salama, A. M. and EL-Sayiad, S. I. 2000. Studies on some sugarbeet cultivars under
middle Egypt conditions: Response to sowing and harvesting dates. Assiut. J.
Agric. Sci., 31(1): 137 - 159.
Arshadi, J. and Asgharipour, M. R. 2011. The effects of seed size on germination and
early seedling growth of pelleted seeds of sugarbeet. Journal of applied
sciences research, 7(8): 1257-1260.
Attia, A.N.; EL-Kassaby A.T.; Badawi M.A. and Seaadh. S.E.E. 1999. Yield, yield
components and quality of sugarbeet as affected by growth regulators, nitrogen
fertilization and foliar nutrition treatments. Proc. 1st Intern. Conf. on Sugar and
Integrated Industries “Present & Future”, 15-18 Feb. 1999, Luxor, Egypt, I: 236-
256.
El-Maghraby, S.; Samia, M.M.; Yusreya, H.T. 1998. Effect of foliar application of nitrogen
and potassium fertilization on sugarbeet. Egypt J. Agric. Res., 76: 665-678.
EL-Shafai, A.M.A. 2000. Effect of nitrogen and potassium fertilization on yield and quality
of sugarbeet in Sohag. Egypt. J. Agric. Res., 78(2): 759-767.
EL-Shahawy, M.I.; EL-Wahab, S.A.A.; Sobh, M.M. and Nemeat, A.E.A.E. 2002.
Productivity and NPK uptake of sugarbeet as influenced by N, B and Mn
fertilization. J. Agric. Sci. Mansoura Univ., 27(3): 1955-1964.
FAO (Food and Agriculture Organization) 2010. Production Year Book. Food and
Agriculture Organization, Roam, 54: 71-79.
Johanson, R.T.; John, T. A.; George, E. R. and George, R. H. 1971. Advances in sugar
beet production, principles and practices. The low State University Press, Ames,
Iowa, USA.
Kashem, M. N. 2014. Effect of sowing date, spacing, nitrogen and potassium on growth,
yield and quality of tropical sugarbeet. Ph.D. thesis, Dept. Agron. BSMRAU.
Gazipur, Bangladesh. pp. 168-176.
Mahasen Fahmi, M. M. 1999. Effect of levels and times of nitrogen application on growth
and yield of sugarbeet. Ph. D. Thesis, Fac. of Agric., Mansoura Univ.
Germination and Yield of Tropical Sugarbeet as .......... Fertilizer 27
Nemeat Alla, E. A. E.; Mohamed, A. A. E. and Zalat, S. S. 2002. Effect of soil and foliar
application of nitrogen fertilization on sugarbeet. J. Agric. Sci. Mansoura Univ.,
27(3): 1343-1351.
Nenadić, N.; Nedić, M.; Živanović, Lj.; Kolarić, Lj. and Gujaničić, T. 2003. Effect of
genotype on sugarbeet yield and quality. Journal of Agricultural Sciences, 48 (1):
1-9.
Ouda, M.M.S. 2002. Effect of nitrogen and sulpher fertilization levels on sugarbeet in
newly cultivated sandy soil. Zagazig J. Agric. Res., 29(1): 33-50.
O'shea, C.J.; Lynch, B.; Lynch, M.B.; Callan, J. J. and O'Doherty, J. V. 2009. Ammonia
emissions and dry matter of separated pig manure fractions as affected by crude
protein concentration and sugarbeet pulp inclusion of finishing pig diets. Agric.
Ecosyst. Environ., 131:154-160.
Ramadan, B.S.H.; Hassan, H.R. and Abdou, A.F. 2003. Effect of mineral and biofertilizers
on photosynthetic pigments, root quality, yield components and anatomical
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Agric. Sci. Mansoura Univ., 28(7): 5139-5160.
Smit, A.B.; Struik, P.C.; Niejenhuis, J.H. and Niejenhuis, V.J.H. 1995. Nitrogen effects in
sugarbeet growing: A module for decision support. Nether lands J. Agric. Sci.,
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as influenced by fertilizer band depth and nitrogen rate in strip tillage . Journal
of Sugarbeet Research, 48 (3 & 4): 137-154.
Bangladesh J. Sugarcane, 35 : 28-36 June, 2014
Genetic Variability, Correlation and Path Analysis of Some
Yield Components of Sugarcane Genotypes
R. Alam1*, M.A. Rahman1, K.M.R. Karim1, H.M. Tarique1 and A.C. Deb2
2 Department of Genetic Engineering and Biotechnology, Rajshahi University,
Bangladesh
1 Breeding Division, Bangladesh Sugarcrop Research Institute
Ishurdi-6620, Pabna, Bangladesh
ABSTRACT
Genetic variability, correlation coefficient, direct and indirect effects
among cane yield and its component traits were estimated for 10 sugarcane
genotypes grown under three environments in two consecutive years. The
genotypes were developed from the crosses of North Carolina Design-I at
Bangladesh Sugarcrop Research Institute. The mean squares for genotypes
were highly significant for all the characters measured. Maximum phenotypic
variation was recorded for cane stalk height (431.01) followed by leaf length
(54.04). Phenotypic and genotypic coefficient of variation was highest for cane
stalk height and germination percentage with value of 164.33 and 13.55,
respectively. The error coefficient of variation was lower than phenotypic but
higher than genotypic coefficient of variation for all the traits. The studied
characters showed low heritability (h2b) ranged from 1.11 to 18.48 and most of
them were positively correlated at genotypic level. The high direct effect of
number of millable canes per clump, leaf breadth both at phenotypic and
genotypic level and germination percentage at phenotypic, cane stalk girth at
genotypic level suggested that these traits are good yield enhancing indices.
Key words: Correlation coefficient, direct and indirect effects, phenotype,
sugarcane
INTRODUCTION
Yield in a crop is one of the most important and complex trait, improvement of
yield remains the top priority in most breeding programme Cox et al. (1994). The
sugarcane varieties trend to decrease its yield after a few years in a particular area. This
necessitates a continuous flow of new varieties from the breeders to maintain productive
genotypes in the field. For a successful sugarcane breeding programme it is important to
know which traits give the highest estimates of heritability and which are the most
repeatable over a number of seasons. Various traits are accounted for variations in cane
and sugar yields. Many of these character components are quantitatively inherited and
interrelated with each other. Stevenson (1965) pointed out that there may not be specific
genes controlling the complex characters, but sum total effect of its component might be
influencing the two most important characters, namely, cane yield and sucrose in
sugarcane. Knowledge of interrelationship among the various characters is considered to
* Corresponding author: R. Alam, Senior Scientific Officer
e-mail: [email protected]
Genetic Variability, Correlation and Path Analysis of Some ........... Genotypes 29
be important in devising proper selection strategies in sugarcane breeding (De Sousa
and Scott, 2005). In the context of yield enhancement, in order to have a good choice of
character for selection of desirable genotypes under planned breeding programme. The
association of component characters with yield and their exact contribution through direct
and indirect effects are very important. Barber (1916) made the first systematic effort in
correlating some of the morphological characters with juice quality in sugarcane.
Correlation coefficient analysis has been extensively used by plant breeders to obtain
precise information on interrelationship among plant traits to better assess outcome of
selecting one or more trait (Furtado et al., 2002). It is a handy technique which provides
information that selection for one character results in progress for other positively
correlated characters. Path coefficient analysis has been widely used in crop breeding to
determine the nature of relationships between grain yield and its contributing
components, and to identify those components with significant effect on yield for potential
use as selection criteria (Samonte et al., 1998). Keeping these views in mind the present
study was aimed with the following objectives:
1. Determine the relationship between cane yield and related characters
2. Find out the direct and indirect effects of morphological traits on cane yield
MATERIALS AND METHODS
Fifteen sugarcane varieties/ genotypes including five males (Isd 40, Isd 35, I
101-66, Co 642 and I 17-01) and 10 females (Isd 31, Isd 29, Isd 25, I 4-71, I 157-94, I
216-92, I 34-95, I 324-86, Co 1148 and CPI 85-80) were mated as North Carolina Design
I (NCD I) of Comstock and Robinson (1952). Following this NCD I each male mated with
two different females and produces 10 progeny families. Each family consists of randomly
selected five F1. The mean values of selected F1 of a family was considered as a
genotype’s value and calling them genotype1 (G1) to genotype10 (G10) for 10 families.
These ten F1 genotypes were subjected in this study as materials.
Experimental site and Design
The whole crossing work was done at Breeding Division of Bangladesh
Sugarcrop Research Institute (BSRI), Ishurdi, Pabna, Bangladesh in the cropping year of
2006 -2007 following North Carolina Design I (NCD I) under a higher study programme.
This crossing design produces 10 progeny families. Each family consists of randomly
selected five F1’s. The mean values of selected F1 of a family was considered as a
genotypic value and calling them genotype 1 (G1) to genotype 10 (G10) for 10 families.
The field trials of these genotypes (G1 - G10) were conducted under three different
locations in two consecutive years 2008-2009 and 2009-2010 following RCB Design with
three replications. The locations were 1. BSRI, Ishurdi, Pabna, 2. Horian, Rajshahi and 3.
Regional Sugarcrop Research Station, Thakurgoan districts of Bangladesh. These
locations are geographically situated at 24°, 24.37° and 26.03°N latitude, 89.25°, 88.6°
and 88.47° E longitude and 30, 24.37 and 37 m above sea level, respectively. The plot
size of the experiment was 4 m × 4 m having a 1 m row-to-row distance. Fertilizers were
applied according to the fertilizers recommended guides of Bangladesh Agricultural
Research Council (BARC), 2005.
30 Bangladesh J. Sugarcane, 35 : 28-36 June, 2014
Data collection and analysis
Data were collected following nine agronomical characters such as germination
percentage (G%), number of tillers/clump (NT/C), number of millable canes/clump
(NMC/C), cane stalk height (CSH), cane stalk girth (CSG), leaf length (LL), leaf breadth
(LB), field brix percentage (Brix%) and cane yield per clump (CY/C). Data of G% and
NT/C were collected 60 and 150 days after plantation, respectively and rest of the data
were collected at the time of harvesting of cane. Data for all the variables measured were
subjected to Analysis of Variance (ANOVA), to estimate the level of variability among the
sugarcane genotypes following the biometrical techniques of analysis as developed by
Mather (1949) based on the mathematical models of Fisher (1932). Genotypic correlation
coefficients (rg) and phenotypic correlation coefficients (rp) were performed following
Singh and Choudhury (1985). Path coefficients analysis was estimated according to the
method suggested by Dewey and Lu (1959).
Genetic variability RESULTS AND DISCUSSION
The mean squares and estimated genetic parameters of the studied sugarcane
genotypes over two growing seasons under three locations are presented in Table 1. It is
clear that the mean squares for the genotypes were highly significant for all the
vcahrairaanccteers(σ2mg)eaansdureedn.virTohnemepnhteanl ovtayrpiaicncvear(iσa2nec)eco(mσ2pp)onweansts. partitioned into genotypic
Phenotypic variation was
greater than those of genotypic variance for all the characters. The highest phenotypic
variation was observed for CSH with a value of 431.01 and the lowest 0.04 by CSG. The
mhoawgenviteurdehigohf eerntvhiraonnmtheentgaelnvoatyripaincceva(rσia2ne)cefor(σa2gll) the yield contributing characters were
Table 1. This result is in accordance
with the report of Podder (1993), Devagiri et al. (1997) in sugarcane and Hossain (2004)
in soyabean. The highest environmental variance (σ2e) with value of 276.69 was found for
CSH followed by LL and G%, while the lowest value of 0.02 for CSG. The highest PCV
and GCV were recorded for CSH and G% with the value of 164.33 and 13.55,
respectively. Whereas, lowest for CSG with the value of 1.77 and 0.13. This result is in
conformity with Mian and Awal (1979) in sugarcane. The environmental coefficient of
variation (ECV) was lower than PCV but higher than GCV for all the traits. The lowest
genotypic value of the characters NMC/C, CSG, LB and CY/C indicating difficulties in
improving these traits through selection as these are under polygenic control.
In this study all the characters showed greater phenotypic coefficient of
variability than genotypic. Similar result is reported by Samad (1991), Hossain et al.
(2000) and Khan (2009). The difference between PCV and GCV was greater in
magnitude for G%, NT/C, NMC/C, CSH, Brix% and CY/C indicated that environment had
considerable effect on these characters. These results are in conformity with the findings
of Podder (1993) and Chubbey and Richharia (1993). The low variability recorded in the
present study for LB and CSG indicating difficulties in improving of these traits through
selection, it should be based on genetic differences. The low variability in cane length,
cane thickness, brix percent and sucrose percent of sugarcane was reported by Singh et
al. (2002); Venkatachalam et al.(2002); Lourdusamy and Selvan (2009) and Anbanandan
and Saravanan (2010). Broad sense heritability (h2b) was low for all the characters
assessed and ranged from 1.11 to 18.48 (Table 1). The low GCV and broad sense
Genetic Variability, Correlation and Path Analysis of Some ........... Genotypes 31
heritability coupled with low genetic gain were observed for all of the characters under
study indicating predominance of non-additive gene under polygenic control and difficulty
in selection. These results are in conformity with Mukopadhya et al. (1986) and Geeta
and Prabhakaran (1987).
Table 1. Estimates of mean squares, variance components and heritability of ten
sugarcane genotypes average over two cropping seasons at three
different locations
Traits Mean MS σ2p σ2g σ2e PCV GCV ECV h2b
118.77** 24.45 4.52 10.99
G% 33.36 73.29 13.55 32.97 18.48
NT/C 5.36 13.61** 0.83 0.009 0.50 15.57 0.17 9.33 1.11
NMC/C 3.61 4.86** 0.48 -0.01 0.29 13.49 -0.34 8.22 -2.54
CSH 262.27 5000.25** 431.01 24.10 276.69 164.33 9.19 105.49 5.59
CSG 2.61 0.56** 0.04 0.003 0.02 1.77 0.13 1.04 7.68
124.29 407.39** 54.04 -8.56 28.28 43.47 -6.89 22.75 -15.85
LL 3.34 0.57** 2.58 -0.01 0.05 2.58 -0.01 1.64 -0.56
LB 18.49 17.74** 1.13 0.56 6.15 0.36 3.05 5.96
Brix% 2.70 4.76** 0.21 0.06 0.11 7.99 0.45 4.10 5.70
CY/C 0.01
MS = mean squares, 2p = Phenotypic variance, 2g = Genotypic variance
2e = Environmental variance
PCV = Phenotypic coefficient of variation, GCV = Genotypic coefficient of variation
ECV = Environmental coefficient of variation and h2b = Heritability in broad sense
Correlation
The association between any two characters is dependent upon their inheritance.
If they are inherited together, the relationship between them may be observed. Genes
governing two or more characters, that is, location of genes on the same chromosome
pair is the cause for association between characters at phenotypic and genotypic levels.
In the present research nine characters were studied and obtained 36 pairs of correlation
coefficients combinations in each case of phenotypic and genotypic level. The phenotypic
and genotypic correlation coefficients among the various characters are presented in
Table 2. The results from the table revealed that cane yield per clump (CY/C) positively
correlated with G% (0.89 and 0.10), NT/C (0.17 and 0.12), NMC/C (0.67 and 0.08), CSH
(0.15 and 0.09) and CSG (0.06 and 0.34) both at phenotypic and genotypic level,
respectively. It suggests that when G%, NMC/C and CSH is increased cane yield will be
increased. Similar result was reported by Kundu and Gupta (1997) and Kadian et al.
(2006). Conversely, CY/C was negatively correlated with LL at phenotypic level and LB
and Brix% at both the levels. The results from the Table 2 revealed that most of the
characters showed positively higher correlation at genotypic level than phenotype.
Highly significant correlation was observed between Brix% and LL and LB at
phenotypic levels indicating Brix% will be increased with an increased leaf length (LL)
and leaf breadth (LB). Long and wide leaf can absorb more sunlight which increases the
photosynthetic rate of the plant to produce more energy that stored in stem as sucrose.
This result is in agreement with Tyagi and singh (2000). Where they observed that
increased sucrose % is related with an increased number of green leaves and top weight,
however they found significant and positive association between pol% cane and
32 Bangladesh J. Sugarcane, 35 : 28-36 June, 2014
sucrose% and top weight. Kumar and Kumar (2014) also reported similar result in
sugarcane.
Table 2. Phenotypic and genotypic correlation coefficients between yield and yield
contributing characters of randomly selected ten sugarcane genotypes
Traits NT/C NMC/C CSH CSG LL LB Brix% CY/C rp
G% 0.25 -0.18 -0.14 -0.89** 0.22 -0.13 0.89** rg
NT/C 0.70 0.59 0.34 0.69 1.58* 0.10 rp
NMC/C 0.91** 0.33 0.36 0.39 -0.93** 0.11 0.17
CSH 0.24 0.06 -0.12 -0.66* 0.94** 1.26** 0.12 rg
CSG 0.22 0.50 -0.14 -0.29 0.67* rp
LL 1.60** 1.03** 0.55
LB 0.18 0.10 -1.03 -0.67* 0.08* rg
Brix% 1.17 0.46 -0.02 0.15 rp
-0.86* 0.41 -0.66* 1.26* 0.09** rg
-0.27 -1.19 -0.41 0.15 0.06
0.88* -1.69* 0.88** 0.70** 0.34 rp
0.06 0.86 1.12** -0.09 rg
1.01** -1.15 0.68**
0.46** -0.04* rp
-0.90 -0.63 rg
-0.21**
-1.05 rp
rg
rp
rg
Here, r>1.0, may be due to subtraction effects arising from sampling error
* and ** indicates significant at 5% and 1% level, respectively
Path analysis
Path coefficient analysis among all characters related to CY/C derived from F1
crosses was estimated separately at phenotypic and genotypic levels. The direct, indirect
and total effects of eight characters on average CY/C over two seasons are presented in
Table 3.
Germination percentage (G%)
The direct effect at phenotypic level of this character was positive (0.63) and
exhibited highly significant positive association with CY/C. In this level the contribution of
other characters indirectly on cane yield through G% appeared to be positive value in
respect of NT (0.09), CSH (0.04), CSG (0.02), LL (0.11) and LB (0.13). The contribution
had negative indirect effects via NMC (-0.16) and Brix% (-0.008). In case of genotypic
level the direct effect of G% (0.17) and other characters those are indirectly involved
on yield character via G% except NT/C (-0.80) and Brix% (-2.55) were positive. In this
level the total effects of G% was positive (0.89 and 0.10) at phenotypic and genotypic
levels, respectively. The total effect of G% at phenotypic level was higher than others but
at genotypic level it was lower than LL and CSG.
Number of tillers per clump (NT/C)
The NT/C expressed a considerably positive direct effect on cane yield (0.37) at
phenotypic level but it was negative (-1.12) at genotypic level. This character had positive
correlation with CY/C at phenotypic and genotypic levels. The indirect effect of NT/C was
positive through NMC/C (0.22) and LL (0.08) and negative via CSH (-0.10), CSG
Genetic Variability, Correlation and Path Analysis of Some ..........Genotypes 33
(-0.007), LB (-0.55) and Brix% (-0.007) at phenotypic level. The association recorded
positive indirect effects via NMC/C (1.56), CSH (0.16), CSG (0.42), LL (0.17), LB (0.80)
and negative via Brix% (-2.04) at genotypic level.
Number of millable canes per clump (NMC/C)
This character appeared to influence sugarcane yield directly as positive value of
0.89 and 0.97 at phenotypic and genotypic level, respectively. NMC/C character recorded
significant positive correlation with CY/C at both the levels. The contribution of other
characters indirectly on cane yield through NMC/C to be positive value in respect of CSH
(0.22 and 0.18) and LL (0.55 and 1.17) and negative via LB (-0.14 and -1.03) and Brix%
(-0.29 and -0.67) at phenotypic and genotypic level, respectively. Therefore this trait
appears to be most important in influencing cane yield and selection should be oriented
towards higher number of millable cane.
Cane Stalk Height (CSH)
The direct influence of cane stalk height towards cane yield (CY/C) was positive
(2.55) at genotypic level but at phenotypic level it was negative (-0.29). The CSH
recorded positive correlation with CY/C at both the levels; in case of genotypic level it
was highly significant. The contribution of other characters indirectly on CY/C through this
character showed positive value in respect of NT (0.12), NMC/C (0.20), LB (0.27) and
Brix% (0.001) at phenotypic level and G% (0.10), NMC/C (0.18) and CSG (0.36) at
genotypic. The contribution showed negative indirect effects via G% (-0.09) and LL (-
0.05) at phenotypic level and NT (-0.07), LL (-0.41), LB (-0.57) and Brix% (-2.04) at
genotypic level.
Cane Stalk Girth (CSG)
It was observed that this character appeared to influence cane yield (CY/C)
directly as positive value 0.06 and 0.41 at phenotypic and genotypic level, respectively.
CSG recorded non significant positive correlation with CY/C at both the levels. The
contribution of other characters indirectly on CY/C through CSG as positive in respect of
G% (0.21), NMC/C (0.08) and CSH (0.08) at phenotypic and G% (0.06), CSH (2.26),
CSG(0.41), LL (0.02) and LB (0.75) at genotypic level. The association exhibited negative
indirect effects via NT (-0.04), LL (-0.08), LB (-0.24) and Brix % (-0.01) at phenotypic and
NT (-1.16), NMC/C (-0.84) and Brix% (-1.15) at genotypic level.
Leaf length (LL)
The direct effect of LL on cane yield (CY/C) was positive (0.34) at genotypic level
and negative (-0.12) at phenotypic level. LL character recorded highly significant positive
correlation with cane yield at genotypic level but it was negatively non-significant at
phenotypic level. The indirect contribution to CY/C through LL was positive in respect of
CSG (0.04) and LB (0.51) at phenotypic level, in case of genotypic levels G% (0.07),
NMC/C (1.15), CSG (0.02), LB (0.86) and Brix% (1.85) were positive. The association
exhibited negative indirect effects via G% (-0.56), NT/C (-0.25), (NMC/C (-0.59), CSH (-
0.12), LL (-0.12) and Brix % (-0.07) at phenotypic level, while CSH ( -3.06) at genotypic
level.
34 Bangladesh J. Sugarcane, 35 : 28-36 June, 2014
Leaf breadth (LB)
The direct contribution of leaf breadth (LB) on cane yield (CY/C) was positive
0.59 and 0.85 at phenotypic and genotypic level, respectively, although LB showed
negative association with cane yield. The contribution of other characters indirectly on LB
appeared to be positive value in respect of G% (0.14) at phenotypic, and G%, CSG, LL
and Brix% at genotypic level. The association exhibited negative indirect effects via NT/C,
NMC/C, and CSH at both the levels, otherwise, CSG, LL and Brix% at phenotypic level.
Table 3. Path-coefficient analysis showing direct and indirect effects of yield
components on yield of sugarcane genotypes at phenotypic and
genotypic levels
Characters G% NT/C NMC/C CSH CSG LL LB Brix% Total
effect
G% 0.63 0.09 -0.16 0.04 0.02 0.11 0.13 -0.008 0.89 P
NT/C 0.17 -0.80 0.89 1.51 0.14 0.13 0.59 -2.55 0.10 G
NMC/C 0.16 0.37 0.22 -0.10 -0.007 0.08 -0.55 -0.007 0.17 P
CSH 0.12 -1.12 1.56 0.16 0.42 0.17 0.80 -2.04 0.10 G
CSG -0.11 0.09 0.89 -0.06 0.006 -0.07 -0.08 0.01 0.67 P
0.16 -1.80 0.97 0.48 -0.35 0.41 -0.88 1.09 0.08 G
LL -0.09 0.12 0.20 -0.29 -0.01 -0.05 0.27 0.001 0.15 P
LB 0.10 -0.07 0.18 2.55 0.36 -0.41 -0.57 -2.04 0.09 G
Brix% 0.21 -0.04 0.08 0.08 0.06 -0.08 -0.24 -0.01 0.06 P
0.06 -1.16 -0.84 2.26 0.41 0.02 0.75 -1.15 0.34 G
-0.56 -0.25 -0.59 -0.12 0.04 -0.12 0.51 -0.07 -1.18 P
0.07 -0.56 1.15 -3.06 0.02 0.34 0.86 1.85 0.68 G
0.14 -0.35 -0.12 -0.13 -0.02 -0.24 0.59 -0.03 -0.17 P
0.12 -1.06 -1.01 -1.70 0.36 0.86 0.85 1.45 -0.12 G
-0.08 0.04 -0.26 0.006 0.009 -0.14 0.27 -0.06 -0.21 P
0.28 -1.42 -0.66 3.24 0.29 -0.40 -0.76 -1.61 -1.05 G
Phenotypic Residual effect= 0.44 and Genotypic Residual effect= 0.87
Field brix percentage (Brix%)
It was observed that this character has no direct effect on cane yield as it shows
negative value (-0.06 and -1.61) both at phenotypic and genotypic levels. At phenotypic
level Brix% recorded significant negative correlation with cane yield, whereas, it was
negatively non significant at genotypic level. The contribution of other characters
indirectly on cane yield through Brix% showed positive value in respect of NT/C (0.04),
CSH (0.006), CSG (0.009) and LB (0.27) at phenotypic level and G% (0.28), CSH (3.24)
and CSG (0.29) at genotypic level. The association exhibited negative indirect effects via
G% (-0.08), NMC/C (-0.26) and LL (-0.14) at phenotypic and NT/C (-1.42), NMC/C (-
0.66), LL (0.40) and LB (-0.76) at genotypic level.
Genetic Variability, Correlation and Path Analysis of Some ..........Genotypes 35
In the present study, the path coefficient analysis was performed for cane yield
as a dependent variable. The high direct effect of NMC/C, LB both at phenotypic and
genotypic level and G% at phenotypic, CSG at genotypic level suggested that these traits
are good yield enhancing indices. However, the indirect effect of LB on cane yield was
negative at both phenotypic and genotypic levels. According to Izge et al. (2006) higher
indirect values could most likely be neutralized in most cases by negative indirect effects
via other characters and this can lead to their low and non-significant genotypic
correlations with total yield. Leaf length (LL) was found negative direct effect on cane
yield at phenotypic level. This negative direct effect was counter balanced through
characters like G%, NT/C, NMC/C, CSH, CSG and Brix% was found negative direct
effect on cane yield both at phenotypic and genotypic levels. The positive indirect effect
of brix was found through NT/C, CSH, CSG and LB at phenotypic level and G%, CSH
and CSG at genotypic level.
Thus, on the basis of correlation and path coefficient analysis it can be concluded
that number of millable canes and cane stalk height are most important characters for
cane yield. Correlation and path analysis revealed that while selecting the clones from F1
and subsequent clonal generation, the characters viz., germination percentage, number of
millable canes, cane height, leaf length and brix% must be taken into consideration for
ameliorating overall sucrose and cane yield.
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Bangladesh J. Sugarcane, 35 : 37-47 June, 2014
Genetic Diversity Analysis and DNA Fingerprinting of Ten
Sugarcane Germplasm Using SSR Markers
A.K. Ghose1*, M.K. Hasan2, K. Mahmud1, N. Islam1, M.A. Rahman3, M.S. Arefin4 and
M.A. Hossain5
2Agriculture Development Officer, CSISA BD, CIMMYT- Bangladesh.
1Biotechnology Division, 3On-farm Research Division, 4Physiology and Sugar Chemistry
Division,5 Director (Research)
Bangladesh Sugarcrop Research Institute, Ishurdi-6620, Pabna, Bangladesh
ABSTRACT
The investigation was conducted for genetic diversity analysis and
DNA fingerprinting of ten sugarcane germplasm viz., K84-200, K76-4, K88-65,
K8887, K8892, PS851, PS862, PS863, PS86-10029 and PS81-362 using four
Simple Sequence Repeats (SSR) primers such as SMC687CS, SMC334BS,
SMC119CG and SMC766BS. The four microsatellite primer pairs amplified a
total number of 102 bands from ten germplasm visualized through 8.0%
polyacrylamide gel electrophoresis. The sizes of the amplified bands obtained
from ten germplasm ranged from 80 to 184bp. The highest number of allele
per locus (4.85) and number of genotypes per locus (4.85) were obtained from
the primer SMC766BS followed by SMC687CS (4.75), SMC119CG (2.17) and
SMC334BS (1.64) respectively. Genetic diversity or polymorphism information
content (PIC) per primer pair ranged from 0.85 to 0.90 with a mean of 0.87 for
all loci across the germplasm evaluated. The primer pairs SMC334BS showed
the highest PIC value 0.90 followed by SMC119CG (0.88), SMC766BS (0.88)
and SMC687CS (0.85). The most polymorphic SSR marker was associated
with the highest number of bands detected. The primer pairs SMC334BS were
the most polymorphic marker for ten germplasm with PIC values of 0.90. The
number of germplasm with unique banding patterns distinguished by the four
SSR markers ranged from 20% to 100%. The four SSR markers were able to
discriminate 42.5% of all the germplasm evaluated with unique banding
patterns. The highest linkage distance was recorded from the germplasm K88-
65, K8892, PS862 and PS863compare with other germplasm investigated.
Genetic relationships among the germplasm at the average distance of 11.8
separated the germplasm K88-65 from the others. Germplasm K88-65 and
PS863 were shown to be outliers and distantly related with the rest of the
germplasm. The four primers were able to identify and classify ten sugarcane
germplasm indicating genetic differences among them.
Key words: Germplasm, fingerprinting, SSR markers, genetic diversity,
sugarcane
*Corresponding author: A. K. Ghose, Scientific Officer
e-mail: [email protected]
38 Bangladesh J. Sugarcane, 35 : 37-47 June, 2014
INTRODUCTION
Sugarcane is one of the major sugar producing crops of the world. It is major
cash-cum-industrial crop in tropical and subtropical regions of the world and important
export product in many developing countries (Heinz et al., 1987). It provides cheap food
in the form of “Sugar” and “Goor (Zaggary)” that lend itself for the production of energy and
many byproducts, all of which are of great economic value to both the developing and
developed countries. Sugarcane is the second most important cash crop, especially north
and southern part of Bangladesh.
Sugarcane breeding programs throughout the world have limited genetic diversity
(Berding and Roach, 1987). Modern sugarcane varieties (Saccharum spp., 2n=100-130)
derived from the interspecific crosses between the sugar-producing species Saccharum
officinarum (2n=80) and wild relatives mainly S. spontaneum (2n=40-128) involved only a
few parental clones (Arceneaux, 1965; Price, 1965). Data from various studies on
chloroplasts (cpDNA) DNA and mitochondria (mtDNA) DNA in the clones of Saccharum
spp. and hybrids indicated a narrow cytoplasmic genome for sugarcane (Mangelsdorf,
1983; D'Hont et al., 1994; Al-Janabi et al., 1994). Such a narrow genetic base has failed
to regenerate new higher yielding varieties. Therefore, there is a need to introduce new
genetic resources in the sugarcane breeding program (Chang, 1996). Genetic base
broadening and enhancement of germplasm are major concerns in variety development.
Potential for maximum gains in the breeding program are only realized when a broad
genetic base is present. Thus, it is needed that DNA fingerprinting of sugarcane
germplasm should be done as short-term strategies to determine the level of genetic
diversity in the collection. The information gained can be used in subsequent activities to
broaden the genetic base of the sugarcane germplasm.
Molecular markers are valuable tools in plant breeding programs as well as for
evolutionary and conservation studies. They are accurate, abundant and less affected by
the environment. Simple sequence repeats (SSRs) also known as mircrosatellites
markers based on tandem repeats of short (2-6 bp) DNA sequences (Litt and Lutty,
1989). These DNA sequences are highly polymorphic even among closely related
cultivars due to mutation causing variation in the number of repeating units (Saghai-
Maroof et al., 1994). SSRs can be analyzed by a rapid, technically simple and
inexpensive polymerase chain reaction (PCR) based assay that requires only small
quantities of DNA. Through PCR, different alleles at a locus can be detected by using
conserved DNA sequences flanking the SSR as primers. SSR markers are co-dominant
and can be transmitted in simple Mendelian segregation. Lastly, SSRs are abundant and
uniformly distributed in plant genomes (Lagererantz et al., 1993; Wang et al., 1994,
Akkaya et al., 1995).
In Bangladesh, characterization of sugarcane germplasm based on agronomic
and morphological traits is being practiced. DNA fingerprinting using RAPD markers has
been initiated by Shahnawaz, 2006. This study was undertaken to identify germplasm
using DNA fingerprinting and determine the genetic diversity of ten sugarcane germplasm
of BSRI germplasm bank using microsatellite markers. Microsatellite is important
particularly for germplasm identification, germplasm selection, hybridization, evaluation
Genetic Diversity Analysis and DNA Fingerprinting of Ten ..... Markers 39
and conservation core germplasm collection. The study was undertaken to identify DNA
fingerprints of sugarcane germplasm and determine the genetic diversity and relationship
among the germplasm by clusters analysis.
MATERIALS AND METHODS
The experiment was carried out at the Biotechnology Laboratory, Bangladesh
Sugarcrop Research Institute (BSRI), Ishurdi-6620, Pabna, Bangladesh.
Plant materials
Ten Sugarcane germplasm viz., K84-200, K76-4, K88- 65, K8887, K8892, PS851,
PS862, PS863, PS86-10029 and PS81-362 collected from BSRI germplasm bank were
used as plant materials for DNA isolation.
Sample collection
Meristem cylinder of sugarcane was used as sample. Top of the field grown 8
month old sugarcane was cut and placed vertically in a bucket containing. Then it was
taken in the laboratory. The outer leaf sheaths were removed, leaving inner spindle. Then
the spindle base was cut down into small pieces with sterile scissors and 0.2g was taken
for DNA isolation.
DNA isolation
In the present investigation, Modified Method of Al-janbi et al. (1999) reported by
Hossain et al. (2006), mini-prep method adopted from Shahnawaz (2006) has been
combined and used to isolate total genomic DNA from sugarcane. Isolated DNA was
stored at –200C for future use.
Primers used
Four selected sugarcane microsatellite primers developed by International
Sugarcane Microsatellites Consortium were used to amplify Simple Sequence Repeats of
genomic DNA of ten sugarcane germplasm (Muyco, 2002). The primers were
SMC687CS, SMC334BS, SMC119CG and SMC766BS (Table 1). Primers were
evaluated on the basis of intensity or resolution of bands, repeatability of markers and
consistency within individual and potential to differentiate varieties (polymorphism).
PCR amplification and electrophoresis
PCR amplification was done in an oil-free thermal cycler (Genius, Techne,
Cambridge Limited) following the PCR profile of 94oC for 5 minutes (initial denaturation)
followed by 35 cycles of 1 minutes denaturation at 94oC, 1 minutes annealing at 55oC
and extension at 72oC for 2 minutes. After the last cycle, a final step of 7 minutes at 72oC
was added to allow the complete extension of all amplified fragments. After completion of
cycling programme, the reactions were held at 40C. PCR reactions were performed on
each DNA sample in a 10µl reaction mixture containing 1.0µl of 10 x (Ampli Taq
polymerase PCR buffer), 1µl of 2.5mM dNTPs, 2.25µl each of forward and reverse primer
from 1.0µM working solution, 0.2µl of 5U/µl Ampli Taq DNA polymerase (Bangalore
Genei Pvt. Ltd., India), 3.0µl of 25ng/µl genomic DNA and a suitable amount (0.3µl) of
sterile de-ionized distilled water.
40 Bangladesh J. Sugarcane, 35 : 37-47 June, 2014
After amplification, 2.0µl loading dye was added to the PCR amplified product
and for separation using polyacrylamide gel electrophoresis. In each well, 3.0µl of PCR
amplified product of each DNA sample for each primer was loaded in 8% polyacrylamide
gel (acrylamide and biacrylamide of SRL, India). Electrophoresis was performed at 50
Volts (5-8V/cm) for 2.5 hours. DNA ladder pBR322HaeIII (Bangalore Genei Pvt. Ltd.,
India) for primer pairs SMC687CS, SMC334BS, SMC119CG and SMC766BS was run
along sides the reactions. After electrophoresis, the gel was silver stained and dried at
room temperature. DNA bands were observed on white light box and photographed by
digital camera.
SSR data analysis
Percentage of polymorphic loci (p)
P = (k/n) x 100%, where k is the number of polymorphic loci and n is the total
number of loci investigated.
Average number of alleles per locus (A )
A = ΣAi/n, where Ai is the number of alleles at the Ith locus and n is the total
number of loci investigated.
Average number of alleles per polymorphic loci (Ap )
Ap=ΣApi/np, where Api is the number of alleles at a certain polymorphic locus and
np is the total number of polymorphic loci investigated.
Average number of genotypes per locus (G )
G=Σg/n, where g is the number of genotypes at a certain locus and n is the
number of loci investigated.
Gene diversity (polymorphic information content)
According to Weir (1990) gene diversity = 1 - ΣP2ij, where P2ij is the frequency of
the jth pattern for the marker i and is summed across n patterns. Anderson et al., (1993)
suggest that gene diversity is the same as the polymorphism information content.
Cluster analysis and dendrogram construction
Following electrophoresis, the size of amplification products were estimated by
comparing the migration of each amplified fragment with that of a known size fragments
of molecular weight marker: 100bp DNA ladder and pBR322HaeIII. All distinct bands or
fragments (SSR marker) were thereby given identification numbers according to their
position on the gel and scored visually on the basis of their presence (1) or absence (0),
separately for each individual germplasm and each primer. The scores obtained using all
primers in the SSR analysis were then combined to create a single data matrix. This was
used for estimating linkage distance (D) and constructing a UPGMA (Unweighted Pair
Group Method of Arithmetic Means) Dendrogram among the varieties using computer
program “Statistica”. Linkage distances were computed from frequencies of polymorphic
markers to estimate genetic relationship between the studied ten sugarcane germplasm
using the Unweighted Pair-Group Method of Arithmetic Means (UPGMA) (Sneath and
Sokal, 1973). The Dendrogram was constructed using the “Statistica” computer package.
Genetic Diversity Analysis and DNA Fingerprinting of Ten ..... Markers 41
RESULTS AND DISCUSSION
DNA fingerprinting followed by genetic diversity was assessed in the investigation
of ten sugarcane germplasm (K84-200, K76-4, K88-65, K8887, K8892, PS851, PS862,
PS863, PS86-10029 and PS81-362) using four SSR primers (SMC687CS, SMC334BS,
SMC119CG and SMC766BS).
The sizes of the amplified bands in the ten sugarcane germplasm ranged from 80
to 184bp (Table 2). SSR primer pair SMC687CS revealed band sizes that ranged from
90bp to 172bp; from 80bp to 184bp for primer SMC334BS; from 89bp to 124bp for primer
SMC119CG and band sizes ranging from 82bp to 166bp for SMC766BS. However, the
primer pair SMC119CG identified band sizes that ranged from 102bp to 152bp (Muyco,
2002). This was perhaps due to the sample differences from this investigation. Yang et al.
(1994) pointed out that the range in allele sizes can be influenced by the large number of
samples screened.
All the four SSR primer pairs amplified a total of 102 bands from the ten
germplasm of sugarcane after PCR amplification and 8% polyacrylamide gel
electrophoresis. For each primer pairs, the number of bands varied from 19 to 34 (Figure
1 and 2). The primer pair SMC766BS amplified the highest number of bands (34)
followed by SMC119CG (26), SMC334BS (23) and SMC687CS (19). Taylor and Cordeiro
(2000) showed that the SSR primer pairs SMC36BUQ and SMC334BS produced
identical fingerprints. The highest number of bands (3.40) per variety was amplified from
the primer pair SMC766BS followed by SMC119CG (2.60), SMC334BS (2.30) and the
lowest number of bands per variety was recorded from the primer SMC687CS (1.90).
Due to the polyploidy nature of sugarcane, the SSR markers revealed multiple bands per
locus. At South Africa Sugar Association Experiment Station (SASEX), Natal, South
Africa work on the application of 35 sugarcane microsatellites identified from 1 to 18
alleles per marker across four varieties (Bester, 2000) (Table 2).
The average number of allele per locus and average number of genotypes per
locus for four primers were identical. The highest number of allele per locus and number
of genotypes per locus (4.85) was obtained from the primer SMC766BS followed by
SMC687CS (4.75), SMC119CG (2.17) and SMC334BS (1.64) respectively. The average
number of allele per polymorphic locus was recorded the highest (19.0) from primer
SMC687CS followed by primer SMC766BS (17.0), SMC119CG (8.67) and the lowest
(2.55) was recorded from primer SMC334BS (Table 3).
Genetic diversity or polymorphism information content (PIC) per primer pair
ranged from 0.85 to 0.90 with a mean of 0.87 for all loci across the ten germplasm
evaluated. The Primer pairs SMC334BS showed the highest PIC value (0.90) followed by
SMC119CG (0.88), SMC766BS (0.88) and the lowest PIC value was 0.85 obtained from
the primer pairs SMC687CS (Table 3).
The most polymorphic SSR marker was associated with the highest number of
bands detected. The primer pairs SMC334BS was the most polymorphic marker across
the ten germplasm with PIC value of 0.90 (Table 3). The PIC values are dependent on
42 Bangladesh J. Sugarcane, 35 : 37-47 June, 2014
the genetic diversity of the germplasm under study. A high proportion of closely related
genotypes would have the effect of lowering the PIC values (Garland et al., 1999).
The number of germplasm with unique banding patterns distinguished by the four
SSR markers ranged from 20% to 100%. The most polymorphic markers were the most
discriminant (SMC334BS) and discriminated 100% of the germplasm evaluated. The four
SSR markers were able to discriminate 42% of all the germplasm evaluated with unique
banding patterns (Table 2).
The highest linkage distance (16.0) was recorded between the germplasm pairs
K88-65 and PS862, K88-65 and PS863, K889 and PS863 than other germplasm
investigated. The linkage distance (7.0) was recorded between germplasm pairs K84-200
and K76-4, PS86-10029 and PS81-362. The same linkage distance (8.0) was recorded
between germplasm pairs PS851 and K8887, PS851 and K8892, PS851 and PS863. The
identical linkage distance (9.0) was recorded between germplasm pairs PS851 and
PS81-362. The germplasm pairs K84-200 and K8887, K84-200 and K8892, K84-200 and
PS86-10029, K88-65 and K8887, PS851 and PS862, PS851 and PS86-10029, PS862
and PS86-10029, PS863 and PS-10029 showed similar linkage distance (10.0). The
linkage distance (11.0) was recorded between germplasm pairs K84-200 and PS81-362,
K76-4 and K88-65, K76-4 and K8887, K76-4 and K8892, K76-4 and PS862, K8887 and
PS81-362, PS862 and PS81-362, PS863 and PS81-362. The germplasm pairs K84-200
and K88-65, K84-200 and PS851, K84-200 and PS862, K8865 and K8892, K8887 and
PS862, K8892 and PS862, PS862 and PS863 showed linkage distance (12.0). The
linkage distance (13.0) was recorded between germplasm pairs K76-4 and PS851, K76-4
and PS863, K76-4 and PS86-10029, K8865 and PS81-362, K8892 and PS81-362. The
linkage distance (14.0) was recorded between germplasm pairs K84-200 and PS863,
K76-4 and PS81-362, K88-65 and PS851, K88-65 and PS86-10029, K8887 and PS863,
K8887 and PS86-10029, K8892 and PS86-10029. The lowest linkage distance (4.0) was
observed between germplasm pairs K8887 and K8892 (Table 4).
Genetic relationships among the germplasm at the average distance of 12.5
showed two major clusters (C1 and C2). At the linkage distance of 11.8 the major cluster
C1 produced sub-cluster SC1 and SC2. Sub-cluster SC2 produced sub-cluster SC3 and
SC4 at the linkage distance of 11.2. At the linkage distance of 11.0 the major cluster C2
produced sub-cluster SC5 and SC6. Sub-cluster SC6 produced sub-cluster SC7 and SC8 at
same linkage distance 10.5. At the linkage distance (8.0) dividing Sub-cluster SC3
produced sub-cluster SC9 and SC10. Similarly, sub-cluster SC4 produced sub-cluster SC11
and SC12 (7.0) and sub-cluster SC8 produced SC13 and SC14 (7.0). Finally, sub-cluster
SC10 divided into two sub-cluster of germplasm K8887 and K8892 at the linkage distance
of 4.0. The sub-cluster SC1 separated the germplasm K88-65 from the other germplasm.
The results of DNA polymorphism of the ten germplasm bear the genetic diversity of
germplasm K88-65 from other germlasms investigated. Germplasm K88-65 and PS863
were shown to be outliers in the dendrogram and distantly related with the rest of the
germplasm based on their genetic distances. The germplasm PS863 lies in the sub-
cluster SC5 and separated from the other germplasm at the linkage distance of 11.0
(Figure 3).
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BJS vol. 35
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