BJS vol. 35

Genetic Diversity Analysis and DNA Fingerprinting of Ten ..... Markers 43

Results of the present investigation revealed that the four SSR primers were able
to identify and classify ten sugarcane germplasm indicating genetic differences among
germplasm. Therefore, DNA fingerprinting and diversity analysis of all the germplasm as
well as entire germplasm collection should be done in order to determine their genetic
relationships among them.

Table 1. Parameters of primers sequences of four sugarcane microsatellite
primers from the International Sugarcane Microsatellite Consortium

Primer Code Sequence G+C Content
SMC687CS Forward: -AGC CAT GCA GGC AGG CAT- (%)
Reverse: -CGC ACA ATC TGC AAG TGC ATC A- 61.11
50.00

SMC334BS Forward: -CAA TTC TGA CCG TGC AAA GAT- 42.85
Reverse: -CGA TGA GCT TGA TTG CGA ATG- 47.61

SMC119CG Forward: -TTC ATC TCT AGC CTA CCC CAA- 47.61
Reverse: -AGC AGC CAT TTA CCC AGG A- 52.63

SMC766BS Forward: -TTA CTC GGC TGG GTT TTG TTC- 47.61
Reverse: -TAA GAA TCG TTC GCT CCA GC- 50.00

Table 2. Microsatellite primers with corresponding bands scored, their size range,
number of polymorphic bands, polymorphism and number of band per
germplasm together with germplasm distinguished in ten sugarcane
germplasm

Size Total Number of Polymorph Number of Germplasm
Primer codes ranges number of polymorphic ism band per distinguished
bands (%) germplasm (%)
(bp) bands
scored

SMC687CS 90-172 19 1 5.26 1.90 20.00

SMC334BS 80-184 23 9 39.13 2.30 100.00

SMC119CG 89-124 26 3 11.54 2.60 30.00

SMC766BS 82-166 34 2 5.88 3.40 20.00
Total - 102 170
Average - 25.5 15 - 42.5

3.75

44 Bangladesh J. Sugarcane, 35 : 37-47 June, 2014

Table 3. Microsatellite primers with corresponding average number of Alleles per
locus, average number of alleles per polymorphic locus, average number
of genotypes per locus together with Polymorphism Information Content

(PIC)

Primer Average Average number Average PIC Mean
codes number of of alleles per number of (Polymorphism PIC
SMC687CS alleles per polymorphic genotypes per 0.87
SMC334BS locus Information
SMC119CG locus 19.00 locus Content)
SMC766BS 4.75 2.55 4.75 0.85
1.64 8.67 1.64 0.90
2.17 17.00 2.17 0.88
4.85 4.85 0.88

Table 4. Summary of linkage distances for different pairs of sugarcane germplasm

Germplasm K84- K76-4 K88- K8887 K8892 PS851 PS862 PS863 PS86- PS81-
200 65 10029 362
10.0 10.0 12.0 12.0 14.0 10.0
K84-200 0.0 7.0 12.0 11.0 11.0 13.0 11.0 13.0 13.0 11.0
10.0 12.0 14.0 16.0 16.0 14.0 14.0
K76-4 7.0 0.0 11.0 0.0 4.0 8.0 12.0 14.0 14.0 13.0
4.0 0.0 8.0 12.0 16.0 14.0 11.0
K88-65 12.0 11.0 0.0 8.0 8.0 0.0 10.0 8.0 10.0 13.0
12.0 12.0 10.0 0.0 12.0 10.0 9.0
K8887 10.0 11.0 10.0 14.0 16.0 8.0 12.0 0.0 10.0 11.0
14.0 14.0 10.0 10.0 10.0 11.0
K8892 10.0 11.0 12.0 11.0 13.0 9.0 11.0 11.0 0.0 7.0
7.0 0.0
PS851 12.0 13.0 14.0

PS862 12.0 11.0 16.0

PS863 14.0 13.0 16.0

PS86-10029 10.0 13.0 14.0

PS81-362 11.0 14.0 13.0

M 1 2 3 4 5 6 7 8 9 10

124bp

104bp

89bp
80bp

Figure 1. DNA Fingerprinting of BSRI ten germplasm of sugarcane based on SSR primer
pair SMC119CG through PAGE (M = marker pBR322Haelll, Lane 1 = K84-200,
Lane 2 = K76-4, Lane 3 = K88-65, Lane 4 = K8887, Lane 5 = K8892, Lane 6 =
PS851, Lane 7 = PS862, Lane 8= PS863, Lane 9 = PS86¬-10029 and Lane 10
= PS81-362)

Genetic Diversity Analysis and DNA Fingerprinting of Ten ..... Markers 45

M 1 2 3 4 5 6 7 8 9 10

184bp
124bp

80bp

Figure 2. DNA Fingerprinting of BSRI ten germplasm of sugarcane based on SSR primer
pair SMC334BS through PAGE (M = marker pBR322HaeIII, Lane 1 = K84-200,
Lane 2 = K76-4, Lane 3 = K88-65, Lane 4 = K8887, Lane 5 = K8892, Lane 6 =
PS851, Lane 7 = PS862, Lane 8= PS863, Lane 9 = PS86¬-10029 and Lane 10
= PS81-362)

Tree Diagram for 10 Germplasm of Sugarcane
Unweighted pair-group average
Squared Euclidean distances

K84200
K764

K8887
K8892
PS851
K8865
PS862
PS8610029
PS81362
PS863

2 4 6 8 10 12 14
Linkage Distance

Figure 3. Cluster analysis by unweighted pair group method of arithmetic means (UPGMA)
of BSRI ten sugarcane germplasm based on four SSR markers

46 Bangladesh J. Sugarcane, 35 : 37-47 June, 2014

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Bangladesh J. Sugarcane, 35 : 48-59 June, 2014

Tolerance Mechanism of Some Sugarcane Genotypes
under Flood Stress

M.K. Begum*, M.S. Arefin and M.J. Islam
Physiology and Sugar Chemistry Division
Bangladesh Sugarcrop Research Institute, Ishurdi-6620, Pabna, Bangladesh

ABSTRACT
A pot study was conducted in an artificially created flood in a concrete
water tank at Bangladesh Sugarcane Research Institute during the cropping
seasons of 2013-2014 to screen flood tolerant genotypes. Biochemical analysis
of juice for determination of brix, pol, purity and reducing sugar were done.
Morphological observations on adventitious roots, growth rate and senesced
leaves, green leaves of seven sugarcane genotypes viz., Isd 20, Isd 34, Isd 37,
Isd 38, Isd 39, Isd 40 and I 25-04 were done under flood stress conditions. The
genotype Isd 34 showed the highest tolerance to flooding for 30, 60 and 90 days
under 30 cm depth above pot soil floods. The variety had >40% green leaf after
120 days stress periods. Genotypes Isd 34 and Isd 39 showed tolerant reactions
following 30, 60 and 90 days flood stress periods with > 33% green leaf after 120
days. Genotype Isd 34 produced the highest adventitious root (AR) (842.0
g/plant) followed by Isd 38 (836.0 g/plant). The genotypes Isd 38 and Isd 39
showed highest growth rate. Isd 38 and Isd 34 showed highest brix percent
(20.55, 20.35) and pol percent (14.48, 14.34) followed by Isd 20 (brix 19.05%
and pol 13.78%). Results of this experiment indicated that genotypes clone Isd
34 and Isd 38 performed better under flood stress conditions than other geno-
types for their better survival mechanism under stress condition.
Key words: Sugarcane, morphological observations, biochemical analysis,

anatomical view of adventitious root
INTRODUCTION

Among abiotic stresses, flood is an important stress for sugarcane cultivation in
Bangladesh. It is because of increased cultivation of sugarcane in low lying char areas
prone to periodic inundation by flood water (Begum et al., 2013). The effect of excess
water stress from temporary or continuous flooding has been studied extensively (Scott et
al., 1989; Jackson et al., 1978). Sugarcane root density is greatest near the soil surface
with 60% in the 0 to 30 cm depth, but roots may penetrate to 180 cm in well- drained soils
(Gascho and Shih, 1983; Paz-Vergara et al., 1980). Flooding tolerance depends on
physiological and anatomical changes of the genotypes. Plants develop a suite of
anatomical, morphological and physiological responses in order to deal with partial
submergence imposed by flooding (Colmer and Voesenek, 2009; Striker et al., 2005).
The most common anatomical response is the generation of aerenchyma in tissues

* Corresponding author: M.K. Begum,Principal Scientific officer
e-mail: [email protected]

Mechanism of Tolerance of Some Sugarcane Genotypes under ... Stress 49

(Seago et al., 2005), which facilitates the transport of oxygen from shoots to roots
(Colmer, 2003). At morphological level, usual responses to flooding include adventitious
rooting and increases in plant height and consequently, in the proportion of biomass
above water level (Naidoo and Mundree, 1993; Grimoldi et al., 1999). This also helps to
facilitate the oxygenation of submerged tissues through the aerenchyma tissue (Colmer,
2003) and at physiological level, flooding modifies water relations and plants carbon
fixation. Closing of stomata, with or without leaf dehydration, reduction of transpiration
and inhibition of photosynthesis, are responses that can occur in hours or days,
depending on the tolerance to flooding of each plant species (Striker et al., 2005; Insausti
et al., 2001; Mollard et al., 2008; Mollard et al., 2010). The following sections show the
main plant responses at those levels associated with tolerance to flooding.

One morphological change in sugarcane roots growing under high water tables is
a greater proportion of fibrous to thick roots in the soil layer above the water table (Eavis,
1972; Webster and Eavis, 1972). The reason is probably an adaptation to lower O2
levels. A thin root has a smaller path-length for O2 diffusion to respiring tissue than a
thicker root (Eavis, 1972). Presence of root aerenchyma is a key requisite for sustained
root activity in flooded soil. The roots of all of the 40 sugarcane genotypes examined
contained aerenchyma (Ray et al., 1996; Heyden et al., 1998). In species that are flood
tolerant, aerenchyma formation is usually constitutive, meaning that it requires no
external stimulus, such as flood (Drew, 1997). Glaze et al. (2002) grew nine sugarcane
cultivars under 15 and 38 cm water table depths. They reported a mean yield reduction of
8.3% at the 15 cm water table, but two cultivars had similar yields at both water tables,
and the yield of one cultivar was reduced by 25% at the 15 cm water table. Generally,
sugarcane is not considered a flood tolerant species, but when it exposed to flood
sugarcane produces adventitious roots which contains aerenchyma which helps
sugarcane genotypes to survive under flood stress. In previous studies, germination and
early seedling growth stages were found most susceptible to flood (Miah and Rahman,
2002). In Bangladesh floods occur at millable cane stage when they grown up much to
fight against stress. Mainly this experiment was conducted to observe the mechanism of
tolerance of some genotypes under flood stress condition.

MATERIALS AND METHODS
An experiment was carried out to observe mechanism of tolerance under flood
stress during the cropping seasons of 2013-2014. Sugarcane genotypes Isd 20, Isd 34,
Isd 37, Isd 38, Isd 39, Isd 40 and I 25-04 were grown in plastic pots (2 pots per clone).
One pre-germinated sett cutting was transplanted in each pot. Irrigations and other
cultural practices were done as and when required to all plant in pot for natural growth.
Six months after transplanting two pots of each genotype were placed in a concrete tank
and inundated in running water (30 cm deep above pot soil), while the remaining three
pots per clone were kept as non-flooded controls. Green and dry leaves counts were
taken after 60, 90 and 120 days of inundation. Data on fresh and dry weight of
adventitious roots as well as volume of adventitious roots (ARs) were taken at harvest.
ARs were collected and taken in paper bags of known weight and oven dried at 85°C until
constant weight. Sections of ARs were made with a sharpe blade and viewed under
microscope and photographed. Tolerance rating scale was recorded on greenness of
leaves and other factors recorded. Data were recorded on growth rate at 60, 90, 120

50 Bangladesh J. Sugarcane, 35 : 48-59 June, 2014

days floods. Laboratory analysis of cane juice was done after 11 months of growth. The
cane samples were crushed in a three-roller power crusher. Soluble solids (Brix %) was
determined by brix hydrometer standardized at 20°C and Horne’s dry lead method was
used for sucrose determination using an automatic polarimeter (Bellingham and Stanley
ADP-220®). Juice purity was calculated as the ratio of the sucrose content and corrected
brix reading. Reducing sugars were determined by the method described in Queensland
Laboratory Manual [Bureau Sugar Experiment Stations (BSES, 1970).

The collected data were compiled and analyzed statistically using the analysis of
variance (ANOVA) technique with the help of a computer package program Statistix 10
and the mean differences were compared by least significance difference test at 5% level
of probability.

RESULTS AND DISCUSSION

Morphological observations in different parameter
Green and dry leaves

Significant differences were observed on dry leaf, green leaf and growth rate by
different genotypes Table 1. Isd 34 produced the highest number of green leaves
(44.07%) followed by Isd 38 (40.0%). Highest growth rate was recorded in Isd 38 (1.290
cm day-1) followed by Isd 39 (1.2750 cm day-1). Dry leaf, green leaf and growth rate were
also affected significantly due to flood and control conditions. Dry and green leaves were
affected significantly and growth rate were unaffected due to different days after initiation
on stress condition. Interaction of factor A (variety) and factor B (Flood, Control), factor A
and factor C (Different days after initiation of flood), factor B and factor C, factor A, factor
B and factor C has significant effect on dry leaf, green leaf and growth rate. All the
genotypes under flood condition at different stress period produced higher no. of green
leaf, and showed higher growth rate than in control condition except in I 25-04 which are
in agreement with Tetsushi and Karim (2007) who found that plant height of the flooded
plants was noticeably higher than that of the control plants. It is possible because
sugarcane has constitutive aerenchyma. For this reason when it falls under stress it can
easily survive by using oxyzen which is preserved by aerenchyma cell (Begum et al.,
2013). Aerenchyma formation in the root cortex is the most studied plastic response to
flooding (Seago et al., 2005; Visser et al., 2000; McDonald et al., 2002; Evans, 2003;
Grimoldi et al., 2005; Striker et al., 2007). This aerenchyma tissue provides a continuous
system of interconnected aerial spaces (aerenchyma lacunae) of lower resistance for
oxygen transport from aerial shoots to submerged roots, allowing root growth and soil
exploration under anaerobic conditions (Colmer and Greenway, 2005). It is predictable
that stress from soil flooding on roots also alters shoot morphology because of the close
functional interdependence between both of them. In this way, flooded plants of tolerant
species are often taller than their non-flooded counterparts as a result of increases in the
insertion angles and length of their aerial organs. These responses were well
characterized in the dicotyledonous Rumex palustris by Cox et al. (2003 and 2004)
and Heydarian et al. (2010) among others.

Interaction effects of genotypes to situation and days after plantation on dry leaf,
green leaf and growth rate differed significantly (Table 7). Genotype Isd 34 produced
lowest dry leaves after 60 days under flood stress condition while the highest were
observed on Isd 39 after 120 days in control condition. Highest green leaves were
produced by Isd 34 after 60 days under flood stress condition followed by Isd 38 after 60

Mechanism of Tolerance of Some Sugarcane Genotypes under ... Stress 51

days under flood condition and Isd 39 after 60 days under flood condition while the lowest
were produced by Isd 39 after 120 days in control condition. Highest growth rate was
observed in Isd 39 after 60 days under flood condition followed by Isd 20 after 60 days
flood condition and Isd 37 after 60 days flood condition while the lowest were observed in
Isd 38 after 120 days in control condition.
Adventitious root

Amount of adventitious roots of various genotypes differed significantly due to
flood stress (Table 8). Among all genotype Isd 34 produced highest adventitious roots
(842.0 g plant-1) followed by Isd 38 (836.0 g plant-1). Flooding induces morphological
changes in roots and shoots. In the sugarcane, the formation of adventitious roots is
highlighted as a common response of flood-tolerant species. These adventitious roots,
which have high porosity, help plants to continue with water and nutrient uptake under
flooding conditions, replacing in some way the functions of older root system (Kozlowski
et al., 1984). It is frequent that these adventitious roots are positioned near the better-
aerated soil surface. Following the review by Jackson (2004), there are three
mechanisms for generating these ‘replacement’ root systems: (i) stimulation of the
outgrowth of pre-existing root primordia in the shoot base (Jackson et al., 1981), (ii)
induction of a new root system that involves initiation of root primordia and their
subsequent outgrowth (Jackson and Armstrong, 1999; Shimamura et al., 2007) and (iii)
placing roots at the soil surface involving the re- orientation of the root extension as seen
for woody species by Pereira and Kozlowski (1977) and for herbaceous species by
Gibberd et al. (2001). The two first mechanisms appear to be triggered by ethylene,
which is thought to increase the sensitivity of plant tissues to auxin (Bertell et al., 1990;
Liu and Reid, 1992) (Figures 2 and 3). Cross section of the adventitious roots showed a
lot of aerenchyma cells (Figure 3). These aerenchyma cell preserved a lot of air which
helps the plants to survive under stress condition. Aerenchyma formation in the root
cortex is the most studied plastic response to flooding (Seago et al., 2005; Visser et al.,
2000; McDonald et al., 2002; Evans, 2003; Grimoldi et al., 2005; Striker et al., 2007). This
aerenchyma tissue provides a continuous system of interconnected aerial spaces
(aerenchyma lacunae) of lower resistance for oxygen transport from aerial shoots to
submerged roots, allowing root growth and soil exploration under anaerobic conditions
(Colmer and Greenway, 2005).

Figure 1. Pictorial view of the experiment Figure 2. Formation of adventitious root
conducted under induced flood due to flood stress
stress in a concrete water tank

52 Bangladesh J. Sugarcane, 35 : 48-59 June, 2014

Isd 20 Isd 34 Isd 37

Isd 38 Isd 39 Isd 40

Figure 3. I 25-04
Cross section of adventitious root of some sugarcane genotypes
showing aerenchyma cell.

Table 1. Effects of genotypes on morphological parameter

Name of Genotypes Dry leaf percentage Green leaf percentage Growth rate
(cm day-1)
Isd 20 (V1) 61.700 38.300
Isd 34 (V2) 55.933 44.067 1.0244
Isd 37 (V3) 61.333 38.000 1.0689
Isd 38 (V4) 58.967 40.200 1.2000
Isd 39 (V5) 60.367 38.967 1.2900
Isd 40 (V6) 60.550 38.783 1.2750
I 25-04 (V7) 63.117 36.217 0.8989
LSD (0.05) 0.3950 0.4055 0.8900
0.0557

Mechanism of Tolerance of Some Sugarcane Genotypes under ... Stress 53

Table 2. Effects of situation on morphological parameter

Name of situation Dry leaf percentage Green leaf percentage Growth rate (cm day-1)
Flood (F) 59.800 39.200 1.2019

Control (C) 60.762 39.238 0.9830
LSD (0.05) 0.2111 NS 0.029

Table 3. Effects of days after plantation on morphological parameter

Name of situation Dry leaf percentage Green leaf percentage Growth rate (cm day-1)
60 days 52.655 46.869 1.1424
90 days 61.705 37.748 1.1014

120 days 66.483 33.040 1.0336
LSD (0.05) 0.2586 0.2655 0.0365

Table 4. Interaction Effects of Genotypes x situation on morphological parameter

Genotypes x situation Dry leaf percentage Green leaf percentage Growth rate (cm day-1)
V1F 60.567 39.433 0.8656
62.833 37.167 1.1833
V1C 59.167 40.833 1.2233
V2F 52.700 47.300 0.9144
V2C
60.133 38.533 1.4733
V3F 62.533 37.467 0.9267
V3C
58.133 40.200 1.4667
V4F 59.800 40.200 1.1133
V4C
59.133 39.533 1.4067
V5F 61.600 38.400 1.1433
V5C 59.433 39.233 0.9644
V6F
61.667 38.333 0.8333
V6C 62.033 36.633 1.0133
V7F
64.200 35.800 0.7667
V7C 0.5586 0.5735 0.0788
LSD (0.05)

Table 5. Interaction Effects of Genotypes x days after plantation on morphological
parameter

Genotypes x days after Dry leaf percentage Green leaf percentage Growth rate (cm day-1)
plantation
V1D1 53.717 45.617 1.1950
62.967 36.367 1.3900
V1D2 67.317 32.017 1.2400
V1D3 50.267 49.067 1.1750
V2D1 62.917 36.417 1.2950
67.917 31.417 1.1300
V2D2 53.100 46.900 1.0900
V2D3 62.850 37.150 1.0750
69.150 30.850 1.0417
V3D1 51.567 47.767 0.9850
V3D2 65.867 33.467 0.8500
V3D3 71.917 27.417 0.8350

V4D1 (Contd.)
V4D2

V4D3

54 Bangladesh J. Sugarcane, 35 : 48-59 June, 2014

Genotypes x days after Dry leaf percentage Green leaf percentage Growth rate (cm day-1)
plantation
51.917 47.417 1.4700
V5D1 59.067 39.767 1.2200
V5D2 65.917 33.417 1.1800
V5D3 52.917 46.417 0.9667
V6D1 61.767 37.567 0.8550
V6D2 66.967 32.367 0.8750
V6D3 55.100 44.900 1.1150
V7D1 56.200 43.800 0.9333
V7D2 56.500 43.500 1.0250
V7D3 0.6841 0.7024 0.0965
LSD (0.05)

Table 6. Interaction Effects of situation x days after plantation on morphological
parameter

situation x days after Dry leaf percentage Green leaf percentage Growth rate (cm day-1)
plantation
51.881 47.167 1.2333
FxD1 61.424 37.481 1.2300
FxD2 66.095 32.952 1.1424
FxD3 53.429 46.571 1.0514
CxD1 61.986 38.014 0.9729
CxD2 66.871 33.129 0.9248
CxD3 0.3657 0.3754 0.0516
LSD (0.05)

Table 7. Interaction Effects of Genotypes x situation x days after plantation on
morphological parameter

Genotypes x situation x Dry leaf percentage Green leaf percentage Growth rate (cm day-1)
days after plantation
V1FD1 52.033 46.633 1.6300
60.633 38.033 1.2400
V1FD2 65.633 33.033 1.3500
V1FD3 53.800 46.200 1.1500
62.900 37.100 1.1500
V1CD1 68.300 31.700 1.1300
V1CD2 48.433 50.233 1.0533
59.400 40.600 0.9200
V1CD3 60.700 39.300 0.9200
V2FD1 52.100 47.900 0.8800
V2FD2 63.700 36.300 0.7900
69.000 31.000 0.8300
V2FD3 52.433 46.233 1.4200
V2CD1 61.133 37.533 1.6300
66.833 31.833 1.3700
V2CD2 55.000 45.000 0.9300
V2CD3 64.800 35.200 0.9600
V3FD1 67.800 32.200 0.8900
51.100 48.900 1.2200
V3FD2 62.500 37.500 1.2800
V3FD3

V3CD1
V3CD2

V3CD3
V4FD1
V4FD2

(Contd.)

Mechanism of Tolerance of Some Sugarcane Genotypes under ... Stress 55

Genotypes x situation x Dry leaf percentage Green leaf percentage Growth rate (cm day-1)
days after plantation
68.100 31.900 1.1700
V4FD3 55.100 44.900 0.9600
V4CD1 63.200 36.800 0.8700
70.200 29.800 0.9133
V4CD2 50.533 48.133 1.1200
V4CD3 64.833 33.833 0.9300
70.733 27.933 0.9900
V5FD1 52.600 47.400 0.8500
V5FD2 66.900 33.100 0.7700
V5FD3 73.100 26.900 0.6800
51.233 47.433 1.8200
V5CD1 58.033 39.633 1.2900
V5CD2 65.133 33.533 1.2900
52.600 47.400 1.1200
V5CD3 60.100 39.900 1.1500
V6FD1 66.700 33.300 1.0700
V6FD2 57.400 42.600 0.7600
62.133 36.533 0.9300
V6FD3 66.833 31.833 0.9067
V6CD1 52.800 47.200 1.4700
52.300 47.700 1.1200
V6CD2 53.000 47.000 0.9600
V6CD3 0.9675 0.9933 0.1365

V7FD1
V7FD2
V7FD3

V7CD1
V7CD2

V7CD3
LSD (0.05)

Table 8. Adventitious roots (AR) of BSRI bred sugarcane clones under induced
flood stress condition (pot experiment).

Name of Genotypes Fresh wt. of adventitious root plant -1 (g)
Isd 20 62.0
Isd 34 842.0

Isd 37 54.8
Isd 38 836.0

Isd 39 380.0
Isd 40 315.7

I 25-04 24.5
LSD (0.05) 1.7512

Biochemical Analysis of Juice
All the parameters viz., brix percentage, purity percentage, pol percentage and

reducing sugar (RS) etc. indicating juice quality Table 9 showed significant difference
among different genotypes. Among them Isd 38 produced the highest brix % (20.55),
highest pol % (14.48) and the highest purity (89.54%) followed by Isd 34 (brix 20.25% , pol
14.34%, purity 89.39%). Flood and control condition showed significant difference on brix
and RS and insignificant effect on pol and purity percentage Table 10. All the genotypes
produced higher brix, pol, purity percentage and lower RS in flood condition than in the
control condition except I 25-04 which are in agreement with Hasan et al. (2003) who
grew some sugarcane genotypes under waterloged condition and found that all the
genotypes had higher brix, pol, purity percentage and lower RS in stress than in control
condition.

56 Bangladesh J. Sugarcane, 35 : 48-59 June, 2014

Interaction effects of genotypes to situation on juice quality differed significantly
Table 11. Genotype Isd 34 produced the highest brix % and pol % followed by genoypes
Isd 38 and Isd 20 under flood stress condition. Isd 38 produced comparatively higher brix
and pol percentage both in stress and control condition. Highest purity percentage were
obtained in genotypes Isd 34 and Isd 38 both stress and control conditions. Isd 20 also
showed better purity percentage in control condition. All the genotypes produced
comparatively lower reducing. Isd 40 produced the highest reducing sugar in control
condition.

Table 9. Effects of genotypes on juice quality

Name of Genotypes Brix (%) Pol (%) Purity (%) Reducing sugar (RS)
Isd 20 (V1) 19.450 13.780 88.460 0.4500

Isd 34 (V2) 20.250 14.340 89.390 0.3400
Isd 37 (V3) 17.500 12.50 87.515 0.4300

Isd 38 (V4) 20.550 14.480 89.540 0.4050
Isd 39 (V5) 19.600 13.590 87.020 0.7100
Isd 40 (V6) 18.800 12.825 83.730 0.7100

I 25-04 (V7) 18.850 12.530 85.910 0.2800
LSD (0.05) 0.6344 0.4395 0.9991 0.008619

Table 10. Effects of situation on juice quality

Name of Genotypes Brix (%) Pol (%) Purity (%) Reducing sugar (RS)
Flood (F) 19.443 13.536 87.881 0.5657

Control (C) 19.129 13.334 86.851 0.3843
LSD (0.05) 0.3391 0.2349 0.5341 0.004607

Table 11. Interaction effects of genotypes x situation on juice quality

Genotypes x situation Brix (%) Pol (%) Purity (%) Reducing sugar
(RS)
V1F 20.000 13.020 87.170 0.2500
V1C 18.900 14.540 89.750 0.6500
V2F 20.800 14.680 89.900 0.3300
V2C 19.700 14.000 88.880 0.3500
V3F 18.000 13.000 87.670 0.4100
V3C 17.000 12.000 87.360 0.4500
V4F 20.600 14.600 89.800 0.4000
V4C 20.500 14.360 89.280
V5F 19.800 13.840 89.040 0.4100
V5C 19.400 13.340 85.000 0.6700
V6F 19.000 13.000 85.650 0.7500
V6C 18.600 12.650 81.810 0.3800
V7F 18.700 12.450 85.880 1.0400
V7C 19.000 12.610 85.940 0.3100
LSD (0.05) 0.8971 0.6215 1.4130 0.2500
0.0122

Mechanism of Tolerance of Some Sugarcane Genotypes under ... Stress 57

Observing the tolerance mechanism of the genotypes which were used in this
study we can concluded that all the genotypes showed better performance under stress
than control condition which produced more adventitious roots with spread aerenchyma
cell are suitable for cultivation under flood stress considering their morphological
characters and biochemical analysis except the genotype I 25-04.

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Bangladesh J. Sugarcane, 35 : 60-72 June, 2014

Productivity and Profitability of Onion Seed Crop-
Mungbean Sequential Intercropping with Sugarcane

M.J. Alam1*, M. M. Rahman2, A.K.M.R. Islam1, M.S. Hossain3, M.A. Razzak1, M.S
Rahman1, H. P. Roy1 and S. Islam4
2 Professor, Agronomy Department, Bangladesh Agricultural University, Mymensingh
3 Regional Sugarcrop Research Station, Thakurgaon
1 Agronomy and Farming Systems Division, 4 Soils and Nutrition Division

Bangladesh Sugarcrop Research Institute, Ishurdi-6620, Pabna, Bangladesh

ABSTRACT
An experiment was conducted at the Bangladesh Sugarcane Research
Institute (BSRI) farm, Ishurdi, Pabna for consecutive two years from 2008-2009
to 2009-2010 to find out the growth, yield, quality and economics of onion seed
crop in onion seed crop-mungbean sequential intercropping with paired rows
transplanted sugarcane. Onion seed crop was grown in 40 cm, 30 cm, 24 cm
and 20 cm row spacing, respectively in between two paired rows of cane. In this
trial, onion bulb - mungbean and potato-mungbean treatments were used as
standard check. Results revealed that sequential intercropping practices did not
have any adverse effect on cane yield and juice quality. All the sequential
intercropping treatments showed higher benefit cost ratio (BCR) than the sole
cane crop. Among the treatment combinations, paired rows cane (PRC) + 20 cm
row spacing (6 rows) of onion seed crop- mungbean performed better in respect
of yield, yield contributing characters and economics. Onion seed crop-
mungbean sequential intercropping with paired rows transplanted sugarcane
could be considered as a technology for sustainable sugarcane farming main-
taining soil health.
Key words: Productivity, sugarcane, paired rows cane, intercropping,

onion seed crop
INTRODUCTION

Sugarcane (Saccharum officinarum L.) is one of the major food-cum-industrial
cash crops in Bangladesh and cultivated in more than 90 countries of the tropical and
sub-tropical regions of the world (FAO, 2011). Sugarcane occupies an area of 23.82
million hectares with a total production of 1685.44 million metric tons (FAO, 2012). In
Bangladesh, on an average 4.5 million metric tons of sugarcane is produced annually
from 0.12 million hectares of land (FAOSTAT, 2012). Sugarcane is mainly grown in high
and medium high land under rainfed conditions in the north-west and south-west regions
of the country. The world average yield of sugarcane is 70.76 tha-1 while in Bangladesh it
is about 44 tha-1 (BBS, 2010). Sugarcane cultivation fetches less benefit compared with
other short duration fruit and vegetable crops like papaya, banana, tomato, carrot,
cabbage and cauliflower. Because of this, sugarcane is being replaced by different

* Corresponding author: M.J. Alam, Principal Scientific Officer
e-mail: [email protected]

Productivity and Profitability of Onion Seed Crop-Mungbean ......... Sugarcane 61

profitable short duration crops. Conventionally sugarcane setts are planted in trench
made at 90-100 cm distances. In many cases, plant population establishment is
hampered in this system. To overcome this situation and ensure plant establishment the
system of settling planting at specified distances in the trenches has been developed.
The transplanting of cane ensures optimum plant establishment. The initial crop growth of
the crop is slow and thus inter row spaces is not covered by sugarcane leaf canopy for
the first 120-150 days. The vacant space can be used for intercrop cultivation. In early
stage of growth, some short duration crops viz., vegetables, pulses, oil seeds and spices
can be grown as intercrop in the vacant spaces between two rows of sugarcane. The
intercropping offers an opportunity of increasing land utilization. The traditional
intercropping with single row sugarcane is less remunerative as the growth and yield of
both the component crops are not satisfactory. Under this situation, paired rows system
of cane cultivation has recently been developed that unveils the potential of practicing
intercropping with sugarcane with high profit. In paired rows system of sugarcane
planting, two rows of cane are planted at 60 cm apart rows in a trench leaving 120-140
cm vacant space between two paired rows of cane (Alam et al., 2008). The paired rows
planting patterns are very easy to adopt and no modifications are necessary in the
existing implements (Umrani, 1981). More than one intercrops in sequence can be easily
cultivated with paired rows system to make sugarcane cultivation more profitable (Alam et
al., 2007). Successful intercropping of various crops with sugarcane has been reported
by many researchers (Alam et al., 2007 and Alam et al., 2008). It has been reported that
potato, onion, garlic and cabbage could be cultivated profitably as first intercrops with
sugarcane (Matin et al., 2001). Besides, after harvesting of first intercrop, the possibility
of growing some short duration crops such as leafy vegetables, mungbean and dhaincha
successfully as second intercrop with sugarcane has been explored. Many reports
suggest that mungbean can be used as second intercrop (Hossain et al., 1995). The
possibility of growing a second intercrop under single row system of cane plantation is
restricted due to canopy development of cane after the harvest of first intercrop. On the
other hand, sequential intercropping in paired rows sugarcane is feasible (De and Singh,
1979).

Among all the intercrops, potato, onion, mustard, lentil, cabbage, cauliflower,
carrot etc., are grown with sugarcane. Mustard ranks the first covering about 40-50% of
the total intercropped area in Bangladesh (Hossain, 1984). In another study, potato yield
was increased by 80-100% in paired rows system compared to the single row system.
The adverse effects of mustard intercrop on tiller, millabe cane and cane yield under
paired rows system increased by 80% compared to the single row intercrop (Ahmed et
al., 2007). The yield of onion as intercrop could be increased under paired rows system of
cane plantation by accommodating higher intercrop population compared to the single
row system. It has been reported that the yield of onion under paired rows systems is 4.4
tha-1 while it was 2.25 tha-1 under single row system (Imam et al., 1990 and Miah et al.,
1994). The intercropping onion with paired rows cane showed the highest potential for
increasing the net returns per unit area (562 US$ ha-1) under intercropped systems
(Imam et al., 1990). Compared to other crops, onion exerted least detrimental effect on
the emergence, tiller, millable cane and yield of sugarcane (Hossain, 1984). Higher yield
of cane due to intercropping with onion has also been reported by (Anon., 1979 and
Parashar et al., 1979). Singh and Rai (1996) reported that cane yield was increased by

62 Bangladesh J. Sugarcane, 35 : 60-72 June, 2014

2.28% when onion was intercropped. Rathi and Singh (1979) and Parashar et al. (1979)
reported that juice quality parameters of sugarcane (brix, purity and sucrose percentages)
were not affected by onion intercropping. Due to shallow rooting depth, onion extract
nutrient from an area on the ridges where the sugarcane shoot roots are not reached at
that stage. This minimizes competition for nutrients and water. Moreover, a lower rate of
pest incidence has been reported in intercropped cane with onion (Verma et al., 1981).
Increased net profits due to intercropping of onion have been reported by Behal and
Narwal (1977) and Verma et al. (1981). Khan et al. (1985) have successfully grown
summer mung with 13 tha-1 fresh biomass weight after harvesting potato and
incorporated the biomass of mungbean into the soil to improve soil fertility and
productivity. Intercropping makes the better use of sunlight and water. It shows some
beneficial effects on pest and diseases (Abdullah et al., 2006). In many cases it gives
higher total production, monetary returns and greater resource use efficiently and
increases the land productivity by almost 60 percent. Intercropping also increases
nutritional quality of diet for the farm family (Khan et al., 2005), allows better control of
weeds, increases land equivalent ratio (Imran et al., 2011). In an intercrop community,
the individual plant of one crop exposed to both intra and inter-specific competition
although causing a reduction in individual yield but total productivity per unit area
increases (Gani and Paul, 2005). Therefore, the present study was undertaken to find out
growth, yield, quality and economics of onion seed crop in onion seed crop-mungbean
sequential intercropping with paired rows transplanted sugarcane.

MATERIALS AND METHODS
The experiment was conducted at the Bangladesh Sugarcane Research Institute
(BSRI) farm, Ishurdi, pabna during 2008-2009 and 2009-2010 cropping seasons. The site
represents High Ganges River Flood Plain under Agro-Ecological Zone 11 with medium
high land of typical sandy loam soil. The experiment was set up in randomized complete
block design with three replications. The unit plot size was 8m × 8m. The treatments are
as follows:
T1 - Paired rows cane (PRC) only
T2 - PRC + onion (bulb) - BINA Mung 5 (standard)
T3 - PRC + potato - BINA Mung 5
T4 - PRC + 40 cm row spacing of onion seed crop - BINA Mung 5
T5 - PRC + 30 cm row spacing of onion seed crop - BINA Mung 5
T6 - PRC + 24 cm row spacing of onion seed crop - BINA Mung 5
T7 - PRC + 20 cm row spacing of onion seed crop - BINA Mung 5
Forty five days old sugarcane settlings of variety Isd 38 were transplanted on 20
November 2008 in 2008-2009 and 08 November 2009 in 2009-10 on well prepared
tranches at 60 cm apart paired rows maintaining 45 cm interplant spacing. The intercrops
were grown within vacant wide space of 140 cm between two paired rows of cane. Onion
bulb was planted at 40, 30, 24 and 20 cm apart rows to accommodate 3, 4, 5 and 6 lines
of onion seed crop in the space between two paired rows. The seed rates of onion seed
(for seed) were 500 kg and 1000 kg ha-1 for paired rows intercrop and sole crop. The first
intercrops (onion, potato and onion seed) were sown on 27 November 2008 and 20
November 2009 for 2008-2009 and 2009-2010 cropping seasons, respectively. The
second intercrop variety BINA Mung 5 was sown in 3 lines in between two paired rows of

Productivity and Profitability of Onion Seed Crop-Mungbean ......... Sugarcane 63

sugarcane on 18 March 2009 and 10 March 2010 in 2008-09 and 2009-10 cropping
seasons, respectively. Sole crop of all first and second intercrops were sown in one side
of the main field on the same days as of intercrops. For sugarcane and intercrops,
fertilizers were applied following the recommended rates (BARC, 2005). N, P, K, S and
Zn were applied in sugarcane @ 150, 50, 90, 34 and 3.5 kg ha-1, respectively while N, P,
K, S, Zn and B were applied @ 75, 30, 75, 30, 3 and 0.6 kg ha-1 for onion. Only N, P and
K were applied in mungbean @ 15, 18 and 14 kg ha-1, respectively. Sole crops received
100% of the recommended doses of fertilizers while 60 % of the recommended rate was
applied in intercrops. N, P, K, S, Zn and B were applied in the form of Urea, triple super
phosphate (TSP), muriate of potash (MoP), Gypsum, Zinc sulphate (ZnSO4) and Boric
acid, respectively. Pest management and necessary intercultural operations like weeding
(3 times), mulching (3 times), gap filling (1 time), irrigation (4 times), earthing up (2 times),
tying (1 time) etc. were done according to the recommendation. Data on number of tiller,
number of millable cane, plant height, diameter of cane, number of internodes stalk-1, unit
stalk weight, cane dry matter m-2 and cane yield were recorded. The plant height, weight
of 1000 grain/seeds, grain/seeds yield, straw yield and days to maturity were recorded for
intercrops. The plant height and bulb yield were recorded for onion. Brix (%), pol (%)
cane, purity (%), recoverable sucrose (%) and sugar yield were also recorded. pH value,
organic matter content, total nitrogen, available phosphorus, exchangeable potassium,
available sulphur and available zinc content of post-harvest soil were measured. Cane
equivalent yield of intercrops and the adjusted cane yield were calculated. Total
production cost, gross income, net return, benefit cost ratio and land equivalent ratio were
calculated for economic analysis.

RESULTS AND DISCUSSION

Growth parameters of sugarcane
Cane height, cane diameter and number of internode cane-1 did not differ
significantly by different row spacing of onion seed crop in onion seed crop - mungbeann
sequential intercropping under paired rows system of planting in 2008-09 and 2009-10
cropping seasons. The total biomass of sugarcane significantly differed due to these
treatment combinations in both the seasons. The highest cane biomass yield of 3080 g
m-2 was 2o4b1s0ergvemd -w2 iwthasT2fo(PunRdCw+ithonTio1n(P- aBirIeNdA Mung 5) and the lowest cane biomass
yield of In 2009-10, the highest cane biomass rows cane only) treatments in 2008-09
season. yield (3100 g m-2) was recorded with
mT7-2()PwRaCs + 20 cm row spacing of onion seed crop - BINA Mung 5) and the lowest (2510 g
found with T1 (Paired rows cane only) treatment (Table 1).

64 Bangladesh J. Sugarcane, 35 : 60-72 June, 2014

Table 1. Effect of row spacing on productivity of onion seed crop in onion -
mungbean sequential intercropping on growth parameters of sugarcane
in 2008-2009 and 2009-2010

Treatments Cane height Cane diameter No. of internodes Dry matter
(m) (cm) cane-1 yield (g m-2)
T1 2008-09 2009-10
T2 2008-09 2009-10 2008-09 2009-10 2008-09 2009-10 2410 c 2510 e
T3 2.58 2.12 2.15 2.02 23 22
T4 2.75 2.61 2.30 2.26 26 28 3080 a 3060 a
T5 2.72 2.25 2.28 2.20 25 24 3050 a 2975 d
T6 2.63 2.58 2.25 2.11 25 26 2440 bc 3022 bc
T7 2.67 2.48 1.97 2.12 23 25 2500 b 3000 cd
2.60 2.56 2.17 2.13 25 26 2450 bc 3050 b
Sx 2.67 2.60 2.10 2.16 24 27
2490 bc 3100 a
CV (%) 0.11 0.08 0.05 0.06 0.69 0.95
Level of 18.00 7.32
significance 9.71 8.55 5.91 6.62 6.28 9.31
NS NS NS NS NS NS 1.72 0.61
** **

In a column, figures with similar letters do not differ significantly at 5% level

T1 - Paired rows cane (PRC) only,
T2 - PRC + onion (bulb) - BINA Mung 5 (standard),
T3 - PRC + potato - BINA Mung 5,
T4 - PRC + 40 cm row spacing of onion seed crop - BINA Mung 5,
T5 - PRC + 30 cm row spacing of onion seed crop - BINA Mung 5,
T6 - PRC + 24 cm row spacing of onion seed crop - BINA Mung 5 and
T7 - PRC + 20 cm row spacing of onion seed crop - BINA Mung 5

Yield and yield attributes of sugarcane
Tiller population, number of millable cane, unit cane weight and cane yield were
not significantly differed among the different intercrops combinaton. The highest number
of tiller 222 × 103 ha-1 and 225 × 103 ha-1 were obtained from T2 (PRC + Onion - BINA
Mung 5) treatment in both the years, The highest number of millable cane of 99 × 103 ha-
1 and 108 × 103 ha-1 were obtained from T2 (PRC + Onion - BINA Mung 5) and T7
treatment in 2008-09 and 2009-10 season, respectively. The highest unit cane weight of
0.94 kg was found with T6 (PRC + 24 cm row spacing of onion seed crop - BINA Mung 5)
in 2008-2009 and that of 0.90 kg awnedre9o3b.1ta2inthead-1frwoemreT1obatnadinTed3 in 2009-10 seasons. The
highest cane yield of 91.54 tha-1 from T2 (PRC + Onion -
BINA Mung 5) treatment in 2008-09 and 2009-10 season, respectively (Table 2).

Productivity and Profitability of Onion Seed Crop-Mungbean ......... Sugarcane 65

Table 2. Effect of row spacing on productivity of onion seed crop in onion -
mungbean sequential intercropping on yield and yield attributes of
sugarcane in 2008-2009 and 2009-2010

Tiller Millable cane Unit cane weight Cane yield
Treatments (×103 ha-1) (×103 ha-1) (kg) (tha-1)
2008-09 2009-10
T1 210 212 2008-09 2009-10 2008-09 2009-10 2008-09 2009-10
T2 222 225 96 90 82.88 81.24
T3 218 215 99 106 0.86 0.90 91.54 93.12
T4 97 98 90.31 87.81
T5 220 224 0.92 0.88
T6 217 223 98 103 83.28 89.25
T7 200 219 96 100 0.93 0.90 86.82 88.54
205 223 89 105 83.69 89.63
Sx 91 108 0.85 0.87 84.08 90.10
5.53 2.91
CV (%) 6.40 3.25 3.32 3.13 0.90 0.89 1.32 2.40
Level of NS NS 8.60 7.60 3.81 6.72
significance NS NS 0.94 0.85 NS NS

0.92 0.83

0.04 0.01

11.98 4.48

NS NS

In a column, figures with similar letters do not differ significantly at 5% level

Growth attributes of intercrops
Plant height, biomass yield, straw yield and crop duration of first intercrops
differed significantly among the different treatment combinations except 1000-grain
weight. The highest plant height of 86.50 cm and 89.10 cm were found with the treatment
T7 (PRC + 20 cm row spacing of onion seed crop - BINA Mung 5) while the lowest plant
height of 42.33 cm and 48.00 cm were recorded with T2 (PRC + onion - BINA Mung 5)
tereatment in both the years. The highest biomass yield of 0.96 tha-1 and 1.08 tha-1 were
observed with the treatment T7 (PRC + 20 cm row spacing of onion seed crop - BINA
Mung 5) while the lowest biomass yield of 0.41 tha-1 ahnigdhe0s.5t 3sttrhaaw-1ywieeldreopf r1o.d2u0ctehda-b1 yanTd2
(PRC + onion - BINA Mung 5) in both the years. The
1.35 tha-1 were observed with the treatment T7 (PRC + 20 cm row spacing of onion seed
crop - BINA Mung 5) while the lowest straw yield of 0.51 tha-1 and 0.66 tha-1 were
produced by T3 (PRC + potato - BINA Mung 5) in both the years. The highest 1000-grain
weight of 3.3 g and 3.33 g were recorded in treatment T4 (PRC + 40 cm row spacing of
onion seed crop - BINA Mung 5) and the lowest 1000-grain weight of 3.27 g and 3.3 g
were observed in T7 (PRC + 20 cm row spacing of onion seed crop - BINA Mung 5)
treatment in both the years. The crop duration was lowest of 94 days and 100 days with
treatment T2 (PRC + onion - BINA Mung 5) and the highest duration of 140 days and 141
days were found with T4 (PRC + 40 cm row spacing of onion seed crop - BINA Mung
5),T5 (PRC + 30 cm row spacing of onion seed crop - BINA Mung 5),T6 (PRC + 24 cm
row spacing of onion seed crop - BINA Mung 5) and T7 (PRC + 20 cm row spacing of
onion seed crop - BINA Mung 5) treatment in both the years (Table 3).

66 Bangladesh J. Sugarcane, 35 : 60-72 June, 2014

Table 3. Effect of row spacing on productivity of onion seed crop in onion -
mungbean sequential intercropping on growth and yield attributes of first
intercrops in 2008-09 and 2009-10

Treatments Plant height Biomass yield Straw yield 1000-grain Crop duration
(cm) (tha-1 ) (tha-1) wt. (g) (days)

2008-09 2009-10 2008-09 2009-10 2008-09 2009-10 2008-09 2009-10 2008-09 2009-10

T1 - - - - - - - - - -

T2 42.33 c 48.00 e 0.41 d 0.53 d 0.83 c 0.95 c - - 94 b 100 b

T3 48.00 c 49.67 e 0.66 c 0.75 c 0.51 d 0.66 d - - 96 b 101 b

T4 85.00 b 88.15 b 0.88 b 0.92 b 1.10 ab 1.15 ab 3.30 3.33 140 a 141 a

T5 84.66 b 86.50 c 0.80 b 0.90 b 1.00 ab 1.12 ab 3.28 3.31 140 a 141 a

T6 84.25 b 85.00 d 0.88 b 0.92 b 1.10 ab 1.15 ab 3.29 3.31 140 a 141 a

T7 86.50 a 89.10 a 0.96 a 1.08 a 1.20 a 1.35 a 3.27 3.30 140 a 141 a

Sx 0.30 0.09 0.04 0.01 0.03 0.01 0.17 0.01 0.88 0.86

CV (%) 1.15 0.34 2.47 0.77 7.43 3.08 19.02 0.75 1.97 1.89

Level of ** ** ** ** ** ** NS NS ** **
significance

In a column, figures with similar letters do not differ significantly at 5% level

Biomass yield of second intercrops was not affected significantly due to onion seed crop
in onion - mungbean sequential intercropping with paired rows transplanted sugarcane
system but crop duration was not affected significantly. The crop duration was 70 days in
2008-09 and 72 days in 2009-10 from sowing to harvesting.

Intercrop yield and adjusted cane yield
The overall yield performance of the first intercrops (Onion, Potato, Onion seed)
under paired rows system was found satisfactory. The yield of mungbean as second
intercrops was significantly influenced by the different treatment combinations. Seed yield
of mungbean was satisfactory in both the years. Among the treatment combination the
highest green biomass yield was obtained of 6.7 tha-1 in 2008-09 and 7.50 tha-1 in 2009-
10 with same treatment while the lowest yield was recorded of 6.20 tha-1 in 2008-09 and
7.10 tha-1 with the same treatment T1(Paired rows cane only). Adjusted cane yield is an
important parameter for determining the total yield potentials of the intercropped plot
(cane + intercrop) over the sole cane plot. In the present experiment, the highest adjusted
cane yield of 290.91 tha-1 and 281.07 tha-1 were obtained from the lTo6wteresat tomf e8n2t .(6P6RtCh+a-61
lines onion seed-BINA Mung 5) in 2008-09 and 2009-10 while the
and 81.24 tha-1 were from the T1 treatment (sole cane ), respectively (Table 4).

Productivity and Profitability of Onion Seed Crop-Mungbean ......... Sugarcane 67

Table 4. Effect of row spacing on productivity of onion seed crop in onion -
mungbean sequential intercropping on intercrops, cane equivalent yield
of intercrops and total adjusted cane yield in 2008-2009 and 2009-2010

Intercrop yield (tha-1) Cane equivalent Adjusted cane yield
yield of intercrop (tha-1)
Treatments First intercrop Second intercrop
(tha-1)
T1 2008-09 2009-10 2008-09 2009-10 2008-09 2009-10 2008-09 2009-10
T2 -- - - 82.88 f 81.24 f
T3 82.88 81.24 237.91 c 163.48 c
T4 7.30 8.00 0.48 0.64 146.37 70.36
T5 55.57 51.30 145.88 e 139.11 e
T6 7.63 8.03 0.43 0.61 145.30 129.75
T7 159.00 140.92
0.21 0.23 - - 179.49 179.67 128.58 d 219.00 d
Sx 0.23 0.25 - - 206.83 190.97 245.82 c 229.46 c
0.26 0.32 - - 263.18 b 269.30 b
CV (%) 0.30 0.34 - - -- 290.91 a 281.07 a
Level of
significance -- - - -- 1.32 2.40
--
-- - - 1.56 2.93
** **
-- --

In a column, figures with similar letters do not differ significantly at 5% level

Price of crops (2008-09) Sugarcane: 1760 Tk. t-1, Potato: 10 Tk. kg-1, Onion (bulb): 32 Tk. kg-1,
Onion seed : 1200 Tk.kg-1, Mungbean : 50 Tk. kg-1 and
Mungbean (GM) : 600 Tk. t-1

Price of crops (2009-10) Sugarcane 2160 Tk. t-1, Potato: 10 Tk. kg-1, Onion (bulb): 15 Tk. kg-1,
Onion seed : 1200 Tk. kg-1, Mungbean : 50 Tk. kg-1 and
Mungbean (GM) : 600 Tk. t-1

Cane juice quality and sugar yield
Sugarcane juice quality parameters like brix (%), pol (%) cane, purity (%),

recoverable sucrose (%) and sugar yield (t ha-1) were not significantly in 2008-09 and
2009-10 (Table 5). The brix (%) ranged from 21.50 to 23.03 and 16.33 to 18.30 in 2008-
09 and 2009-2010 seasons. In case of pol (%) cane ranged from 15.34 to 16.07 and
11.10 to 12.88 in 2008-09 and 2009-10 seasons. The purity (%) ranged from 88.62 to
91.68 and 85.89 to 88.95 in 2008-09 and 2009-10 seasons. The recoverable sucrose (%)
ranged from 12.55 to 13.57 and 9.10 to 10.38 in 2008-09 and 2009-10 seasons. Sugar
yield (tha-1) ranged from 9.90 to 11.91 and 7.39 to 8.92 in 2008-09 and 2009-10 seasons.

68 Bangladesh J. Sugarcane, 35 : 60-72 June, 2014

Table 5. Effect of row spacing on productivity of onion seed crop in onion -
mungbean sequential intercropping on juice quality and sugar yield (tha-1)
in 2008 -09 and 2009-10

Treatments Brix (%) Pol (%) cane Purity (%) Recoverable Sugar yield
sucrose (%) (tha-1)

2008-09 2009-10 2008-09 2009-10 2008-09 2009-10 2008-09 2009-10 2008-09 2009-10

T1 21.60 18.30 15.37 12.88 90.71 88.90 12.87 10.38 9.90 7.39

T2 22.10 17.22 15.75 11.96 91.50 88.95 12.55 9.10 11.49 8.47

T3 21.80 16.33 15.69 11.10 91.68 85.89 13.19 8.59 11.91 7.55

T4 23.03 17.80 16.01 12.50 88.62 88.85 13.51 9.99 11.25 8.92

T5 21.50 17.07 15.34 11.81 90.90 87.49 12.84 9.31 11.14 8.24

T6 22.50 16.37 16.07 11.46 91.03 86.69 13.57 8.96 10.82 7.80

T7 22.30 18.23 15.84 12.62 90.53 87.60 13.34 10.12 11.20 8.12

Sx 0.42 0.57 0.20 0.49 0.78 0.78 0.20 0.49 0.23 0.40

CV (%) 4.59 8.03 3.04 9.93 2.11 2.22 3.61 12.53 4.94 11.77

Level of NS NS NS NS NS NS NS NS NS NS
significance

In a column, figures with similar letters do not differ significantly at 5% level

Economic analyses

Cost of cultivation
The cost of cultivation was estimated for different sequential intercropping
treatment. The highest costs of cultivation 1,62,000 Tk.ha-1 was required for the
Tt2r0ke0.ah8tam--01e9nwtaaTns2dr(e1Pq,R6u1Cir,e0+d0o0snoTiloekn.hcbaaun-1lebini-nTBb7INotdAhuMrthinuegnsg2e05a0)s9oa-nn1sd0 T3 (PRC + potato - BINA Mung 5) in
seasons. The lowest cost of 80,000
(Table 6). Other treatments required
1,47,000 Tk.ha-1 and 1,55,000 Tk.ha-1 ainndT14,5tr9e,a0t0m0eTnkt;.h1a,-41 9in,0T060anTdk.1h,a5-13,a0n0d0 1,57,000
Tk.ha-1 in T5 treatment; 1,51,000 Tk.ha-1 Tk.ha-1 in
2008-09 and 2009-10 seasons, respectively.

Gross income and net return
Under intercrop condition, the highest gross income of Tk. 5,12,001 ha-1 and Tk.

l16o0,w07e(T,s1kt1.g6r1o,hs7as5-,1i4n7wc8oamsheao-wb1)taassinefearodsmofnrtoshm,ereTths1petereTcat7itvmtereleyna.ttmSiniem2ni0tla0irn8-r0e29s0u0(lT8t -kh0.a91s,4aa5nl,ds8o629b0e0he9an--11)0obawtnahdinil2ee0d0th9ine-
case of net return in 2008-09 and 2009-10 (Table 6).
Benefit cost ratio (BCR) and land equivalent ratio (LER)

The highest BCR of 3.35 and 3.77 were found from T7 (PRC + 24 cm row spacing
of onion seed crop - BINA Mung 5) treatment in 2008-09 and 2009-10, respectively. The
lowest BCR of 1.58 and 2.07 were observed from T3 (PRC + potato - BINA Mung 5)
treatment 2008-09 and 2009-10, respectively. The highest LER of 2.31 in 2008-09 and
2.59 in 2009-10 from T6 (PRC + 24 cm row spacing of onion seed crop - BINA Mung 5)
treatment while the lowest LER of 1.95 in 2008-09 from T4 (PRC + 40 cm row spacing of
onion seed crop - BINA Mung 5) and 2.27 in 2009-10 was obtained from T3 (PRC +
potato - BINA Mung 5) treatments (Table 6).

Productivity and Profitability of Onion Seed Crop-Mungbean ......... Sugarcane 69

Table 6. Effect of row spacing on productivity of onion seed crop in onion -
mungbean sequential intercropping on cost of production, benefit cost
ratio (BCR) and land equivalent ratio (LER) in 2008-09 and 2009-10

Treatments Total production Gross income Net return BCR LER
cost (Tk. ha-1) (Tk. ha-1)
T1 2008-09 2009-10 2008-09 2009-10
T2 (Tk. ha-1) 2008-09 2009-10 2008-09 2009-10 1.82 2.19 1.00 1.00
T3 145869 175478 65868 95478 2.58 2.44 2.29 2.38
T4 2008-09 2009-10 418721 353116 256721 208116 1.58 2.07 2.20 2.27
T5 80000 80000 256746 300470 94745 155469 2.74 3.05 1.95 2.34
T6 162000 145000 402293 473040 255292 318040 2.90 3.16 2.12 2.39
T7 432643 495626 283643 338626 3.07 3.66 2.31 2.59
162000 145000 463194 581681 312194 422680 3.35 3.77 2.30 2.49
147000 155000 512001 607116 359000 446116
149000 157000
151000 159000
153000 161000

Soil characteristics
Effects of row spacing on productivity of onion seed crop in onion-mungbean

sequential intercropping in paired row transplanted sugarcane on post harvest soil pH,
total N content (%), and available Zn (mg kg-1) were not significant while that on organic
matter (%), available P (mg kg-1), exchangeable K (cmol kg-1) and available S (mg kg-1)
were significant in both the years. The highest organic matter was obtained in T2 (1.07 %
hmAi&2Ani0vvgg10aathr.9iikeell0aa-gas61bb-tt10mll)ee%wweiiSPn)tnhhtwiww2letTh0arae3i0tsslhea9(tett1-hhmt13heel0oe.e2whhnw0iitleggohsThhawitlee5neewssd(stt0tathw1w.s1ew4ii6itt.nahhl2osc0ttwTrrmeei6enmaoas(tl1gtttmmr7kewkeg.ea8gan-nt10-smt)t1)TmTieinni12nngt(2(t0r2kb0eT.g6o01a1-.t1804thm(-0&1a0te.n9hm10nde90gta.0ny0Tka.ed0g11na-2d1m(irn1)scg80i.mTn..kE592o2g07x&l0-c1k0a%)hgT8naw-)7-d1n0h)(ig9n0i2ilene.1a1ab.btn82bohod0lteechthimnmloKttohhwgTlewe4ekkas(yygg2tsee--511waa))t.hrr2assiie0nns..
both the years (Table 7 and 8).

70 Bangladesh J. Sugarcane, 35 : 60-72 June, 2014

Table 7. Effect of row spacing on productivity of onion seed crop in onion -
mungbean sequential intercropping on soil pH, organic matter (%), total
N content (%) and available P (mg kg-1) content of post harvest soil in
2008-09 and 2009-10

Treatments pH Organic Total N Available
matter (%) content (%) P (mg kg-1)
2008-09 2009-10
2008-09 2009-10 2008-09 2009-10 2008-09 2009-10 16.00 15.70
1.00 0.97 0.04 0.04 16.70 18.25
T1 7.6 7.6 1.07 1.06 0.05 0.06 13.20 14.20
T2 7.6 7.7 16.50 18.40
T3 7.5 7.6 1.02 1.02 0.06 0.05 16.50 16.00
T4 7.6 7.7 1.05 1.05 0.05 0.06 17.80 19.00
T5 7.5 7.7 1.06 1.05 0.06 0.06 16.40 15.80
T6 7.5 7.6
T7 7.5 7.7 1.00 1.05 0.05 0.05 0.45 0.47
1.02 1.04 0.06 0.05
6.93 7.07
Sx 0.05 0.05 0.01 0.01 0.01 0.01 ** **

CV (%) 1.55 1.64 0.87 1.80 11.72 19.37
NS NS NS NS
Level of NS NS
significance

In a column, figures with similar letters do not differ significantly at 5% level

Table 8. Effect of row spacing on productivity of onion seed crop in onion -
mungbean sequential intercropping on exchangeable K (cmol kg-1),
available S (mg kg-1) and available Zn (mg kg-1) content of post harvest
soil in 2008-09 and 2009-10 cropping seasons

Treatments Exchangeable K (cmol kg-1) Available S (mg kg-1) Available Zn (mg kg-1)

2008-09 2009-10 2008-09 2009-10 2008-09 2009-10

T1 0.10 0.12 18.50 21.20 0.54 0.60

T2 0.15 0.18 26.40 24.00 0.65 0.66

T3 0.12 0.15 23.30 24.50 0.57 0.65

T4 0.14 0.13 19.50 25.20 0.57 0.64

T5 0.16 0.17 20.00 22.00 0.66 0.62

T6 0.13 0.16 19.50 23.50 0.64 0.66

T7 0.14 0.18 25.50 23.20 0.61 0.65

Sx 0.14 0.01 0.25 0.49 0.01 0.01
2.98 5.31 2.74 2.36
CV (%) 48.94 15.56 NS NS
** **
Levelof ** **

significance

In a column, figures with similar letters do not differ significantly at 5% level

From the experiment it can be concluded that intercropping of onion seed crop with
sugarcane does not have any adverse effect on growth, yield and juice quality of
sugarcane. Onion seed crop intercropping was economically more profitable than sole
cane and the highest economic benefit was obtained from the onion seed crop-

Productivity and Profitability of Onion Seed Crop-Mungbean ......... Sugarcane 71

mungbean sequential intercropping using six rows onion seed crop maintaining 20 cm
row spacing in between two paired rows of transplanted sugarcane. Therefore, this
intercropping sequence can be practiced for sustaining sugarcane farming and
maintaining soil health.

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Bangladesh J. Sugarcane, 35 : 73-78 June, 2014

Response of 2, 4-D on Callus Induction and Plantlet
Regeneration in Sugarcane

K. Mahmud1*, K.M. Nasiruddin2, M.A. Hossain3 M.A. Rahman4 and A.K. Ghose1
2Prof. Dept. of Biotechnology, Bangladesh Agricultural University, Mymensingh
1Biotechnology Division, 4On-farm research Division, 3Director (Research)
Bangladesh Sugarcrop Research Institute, Ishurdi-6620, Pabna, Bangladesh

ABSTRACT
The experiment was conducted at the Biotechnology Laboratory,
Bangladesh Sugarcane Research Institute (BSRI), Ishurdi, Pabna,
Bangladesh. The effects of only MS medium, MS medium with green coconut
water (10%, 20%, 30%, 40% and 50%) and MS medium with green coconut
water containing various concentration (1.0, 2.0, 3.0, 4.0 and 5.0 mg l-1) of 2,
4-D on callus induction, shoot and root formation of sugarcane variety Isd 40
were investigated. No callus, shoot and root were produced in only MS
medium and MS medium with green coconut water (10%, 20%, 30%, 40% and
50%). But MS medium containing green coconut water supplemented with
different concentrations of 2, 4-D (1.0, 2.0, 3.0, 4.0 and 5.0 mg l-1) produced
callus, shoot and also root. The frequency of callus induction varied from 76.67
to 100 % depending on the different concentrations of 2, 4-D. Among the callus
induction medium containing 3.0 mg l-1 2, 4-D was found to be the best for
callusing (100%). After callus formation a few number of shoots and roots were
regenerated directly from some callus. The highest shoots (7.0 per callus) and
roots (4.0 per callus) were produced from MS medium containing green
coconut water supplemented with 1 mg l-1 2, 4-D. On the other hand, the
lowest shoots (1.33 per callus) were produced from MS medium with green
coconut water containing 5 mg l-1 2, 4-D. Besides, no roots were produced
from MS medium with green coconut water containing 4 and 5 mg l-1 2, 4-D.
The lowest day’s requirement for induction of callus (12.00) and shoot (21.33)
were obtained from MS medium with green coconut water containing 1 mg l-1
2, 4-D. Therefore, it may be recommended that 3.0 mg l-1 2, 4-D may be used
for only callus induction but 1.0 to 3.0 mg l-1 2, 4-D for callusing with shooting
and rooting in sugarcane variety Isd 40.
Key words: Callus, 2, 4 - D, root and shoot regeneration, sugarcane

INTRODUCTION
Sugarcane is globally the main source of raw material for the production of sugar.
It is a principal cash crop in north western and south western low rainfall belt of the
country and the main raw material for sugar and goor industries. The genus Saccharum
contains three cultivated (S. officinarum, S. sinensis and S. barbari L) and two wild (S.
robustum and S. spontanium) species (Kochhar, 1998). The geographical origin of
modern cultivated sugarcane (S. officinarum) is New Guinea and distributed throughout
the tropics and sub-tropics are generally agreed. Sugarcane plant regenerated from
* Corresponding author: K. Mahmud, Principal Scientific Officer
e-mail: [email protected]

74 Bangladesh J. Sugarcane, 35 : 73-78 June, 2014

tissue and cell culture show heritable variation for both qualitative and quantitative trait
and considered as a powerful tool for crop improvement within limited time period. For the
purpose development of regeneration protocols is prerequisite. Leaf tissue of modern
hybrids of sugarcane clones is usually more responsive to tissue culture than those of
traditional varieties or wild relatives (Chen et al., 1998; Taylor et al., 1997). The
sugarcane breeding programme has been under serious problem due to lack of suitable
multiplication procedure (Lal and Sing, 1994). Conventional breeding method usually
required 10-15 years of work to complete a section cycle for release of a variety and an
important variety can be planted commercially several years later when enough seed
canes will have been produced. Time required and continued contaminations by systemic
diseases are serious problem to multiply a-elite genotype of sugarcane in the open field
(Lal and Sing, 1994). The technique of plant tissue culture is being routinely used for
producing large number of clonal plants by in vitro culture of explants from wide range of
species throughout the world. It has become now a viable alternative to the conventional
breeding and the clonal propagation methods. Due to increased demand of sugar and goor
for local consumption, sugarcane is being cultivated years together without adopting
modern technologies. To meet the future requirement of sugar it is essential to develop
some improved varieties by tissue culture. Although, sugarcane is one of the most
important industrial crops, very limited effort have been made on tissue culture and in
vitro propagation for variety development and multiplication of Bangladesh. Hence tissue
culture research using modern sugarcane varieties of Bangladesh deserves due
attention. Therefore, this experiment was conducted to find out direct callusing with
shooting and rooting potentiality of sugarcane variety Isd 40 by applying growth regulator
hormone 2, 4-D.

MATERIALS AND METHODS
The experiment was conducted at the Biotechnology Laboratory, Bangladesh
Sugarcane Research Institute (BSRI), Ishurdi, Pabna, Bangladesh during the period from
September, 2010 to January, 2011 to obtain in vitro plant regeneration potentiality of
BSRI released variety Isd 40 (Figure 1). The experimental materials was the young leaf
sheath of BSRI released variety Isd 40. The explants were collected from 8-10 months
old field grown sugarcane from BSRI experimental field. Only MS medium, MS medium
with coconut water (10%, 20%, 30%, 40% and 50%) and MS medium with green coconut
water (10%) containing five concentrations of 2, 4-D (1, 2, 3, 4 and 5 mg l-1) were used as
treatment for callus induction, shoot and root formation. Mediums were adjusted to pH
(5.8). Agar (0.6%) was added medium. All media were sterilized by autoclaving at 1.2 Kg
cm-2 pressure at 121o C for 30 minutes. Mercuric Chloride (HgCl2) was used as sterilizing
agent while savlon was used as antiseptic, detergent and surfactant. The explants were
taken in a beaker and treated with 1% (w/v) savlon for 5-6 minutes with constant shaking
and washed thoroughly with distilled water for 3-4 minutes. The explants were transferred
in autoclaved conical flask (500 ml) treated with 0.1% (HgCl2) for 10 minutes
and washed by 3-4 times rinsing with sterile distilled water to remove traces of HgCl2 from
outer surface of leaf sheath segments. Explants (approximately 1 cm x 0.5 cm) were
prepared in laminar air flow cabinet from sterilized leaf sheath segments and cultured on
only MS medium, MS medium with coconut water and MS medium with green coconut
water containing five concentrations of 2, 4-D (1, 2, 3, 4 and 5 mg l-1). Cultures were
incubated at 25±2oC and kept 16 hour under fluorescent tube light. The experiment was laid
out in Completely Randomized Design (CRD). Three replications and ten test tubes of

Response of 2, 4-D on Callus Induction and Plantlet ........ Sugarcane 75

of each treatment were maintained for observation. The data for the characters under the
present study were statistically analyzed following CRD. The analysis of variance was
performed by Least Significant Difference (LSD) test at 5% level of probability for
interpretation of result (Gomez and Gomez, 1984).

RESULTS AND DISCUSSION
Different concentrations of 2, 4-D had significant effects on callus, shoot, root
induction etc., (Figure 1 and 2, Table 1 and 2). When explants were inoculated on MS
medium, MS medium with green coconut water (10%, 20%, 30%, 40% and 50%) and MS
medium with green coconut water (10%) supplemented with 2, 4- D (1.0, 2.0, 3.0, 4.0 and
5.0 mg l-1), no callus was produced on both only MS medium and MS medium with green
coconut water (Figure 1). On the other hand, the highest callus (100%) was produced
from MS medium containing green coconut water supplemented with 2, 4 – D 3 mg l-1
followed by MS medium containing green coconut water supplemented with 2, 4–D 4 mg
l-1 (96.67%) (Table 1 & Figure 1). This finding is in agreement with the results of Alam et
al. (2003); Karim et al. (2002); Hossain et al. (1996); Ali et al. (2008); Gopitha et al.
(2010) and Mahmud et al. (2013). The lowest days (12.0) requirement of callus induction
were obtained from MS medium containing green coconut water supplemented with 2, 4–
D 1 mg l-1 after explants inoculation while the highest days (13.67) requirement of callus
induction from MS medium containing green coconut water supplemented with 2, 4–D (4
& 5) mg l-1. Besides, the highest weight (1.77 gm) callus-1 was recorded from MS medium
containing green coconut water supplemented with 2, 4–D 3 mg l-1 where the lowest
weight (1.37 gm) callus-1 from MS medium containing green coconut water supplemented
with 2, 4–D 5 mg l-1. It is revealed that shoots and roots were produced directly from
callus derived from MS medium with green coconut water supplemented 2, 4-D (1.0, 2.0,
3.0, 4.0 and 5.0 mg l-1) (Figure 1). The highest callus % (96.67) produced shoots from MS
medium with green coconut water supplemented 2, 4-D 1.0 mg l-1 followed by MS
medium with green coconut water supplemented 2, 4-D (2.0 & 3.0) mg l-1 (76.67%) while
the lowest (23.33%) from MS medium with green coconut water supplemented with 2, 4-
D 5.0 mg l-1 (Table 2). The highest number of shoots per callus (7.0) were found from MS
medium containing green coconut water supplemented with 2, 4–D (1.0 & 2.0) mg l-1
while the lowest (1.33 shoots per callus) from MS medium with green coconut water
supplemented 2, 4-D 5.0 mg l-1. The lowest days (21.33) requirement of shoot induction
were obtained from MS medium containing green coconut water supplemented with 2, 4–
D 1.0 mg l-1 after callus formation while the highest days (35.67) requirement of shoot
induction from MS medium containing green coconut water supplemented with 2, 4–D 5.0
mg l-1. The highest number of roots callus-1(4.0) were obtained from MS medium
containing green coconut water supplemented with 2, 4–D 1.0 mg l-1 followed by MS
medium containing green coconut water supplemented with 2, 4–D 2.0 mg l-1 (2.0 roots
per callus). On the contrary, no root was produced by MS medium with green coconut
water supplemented with 2, 4-D (4.0 & 5.0) mg l-1 (Table 2). Finally regenerated plantlets
were transplanted in the polybag and then field after hardening (Figure 2).

76 Bangladesh J. Sugarcane, 35 : 73-78 June, 2014

Figure 1. A. Sugarcane variety Isd 40, B. No callus produced in MS medium with
green coconut water (GCW), C. Callus induced from sugarcane variety Isd
40 under concentration of 3 mg/l 2, 4–D, D. Regeneration of shoots
produced from leaf sheath derived calli of sugarcane variety Isd 40 under
concentration of 3 mg/l 2, 4–D supplemented with 10% green coconut water

Response of 2, 4-D on Callus Induction and Plantlet ........ Sugarcane 77

Figure 2. A. Regenerated plantlets were transplanted in the polybag and then
hardening shade, B. Somaclone in the field

Table 1. Effects of 2, 4-D on callus induction in Sugarcane

Treatments Callus induction Days required of Weight (gm)/Callus
1 mg l-1 2, 4-D (%) callus initiation 1.43 c
2 mg l-1 2, 4-D 1.63 b
3 mg l-1 2, 4-D 76.67 b 12.00 1.77 a
4 mg l-1 2, 4-D 80.00 b 12.33 1.63 b
5 mg l-1 2, 4-D 100.00 a 12.67 1.37 c
LSD (0.05) 96.67 a 13.67 0.1238
76.67 b 13.67
5.538
-

Figure in the column with same letter do not differ significantly at 5% level of probability
as per DMRT
Table 2. Effects of 2, 4-D on shoots and roots formation in sugarcane

Treatments Callus % Number of Days required Number of
1 mg l-1 2, 4-D produced shoot/callus of shoot root/callus
2 mg l-1 2, 4-D initiation
3 mg l-1 2, 4-D shoot 7.00 a 21.33 d 4.00 a
4 mg l-1 2, 4-D 96.67 a 7.00 a 21.67 d 2.00 b
5 mg l-1 2, 4-D 76.67 b 5.67 b 23.33 c 1.33 c
LSD (0.05) 76.67 b 4.00 c 24.33 b 0.00 d
53.33 c 1.33 d 35.67 a 0.00 d
23.33 d 0.599 0.5538 0.4533
9.592

Figure in the column with same letter do not differ significantly at 5% level of probability
as per DMRT

78 Bangladesh J. Sugarcane, 35 : 73-78 June, 2014

REFERENCES
Alam, R.; Mannan, S.K.; Karim, Z. and Amin, M.N. 2003. Regeneration of sugarcane

(Saccharum officinarum) plantlet for callus. Pak. Sugar J., 18(1): 15-19.
Ali, A.; Naz, S.; Siddique, F. A. and Iqbal, J. 2008. Rapid clonal multiplication of

sugarcane (Saccharum officinarum) through callogenesis and organogenesis.
Pak. J. Bot., 40(1): 123-138.
Bhansali, R. R. and Kishan, S. 1982. Callus and shoot formation from leaf of sugarcane
in tissue culture. Phytomorphology, pp.167-170.
Burner, D. M. and Grishan, M. P. 1995. Induction and stability of phenotypic variation in
sugarcane as affected by propagation procedure. Crop Science, 35: pp. 875-880.
Chen, W. H.; Davey, M. R.; Power, J. B. and Cocking, E. C. 1998. Control and
maintenance of plant regeneration in sugarcane callus culture. J. Expt. Bot., 39:
pp. 251-261.
Evans, D. A.; Sharp, W. R. and Medmi-Filho, H. P. 1984. Somaclonal and gametoclonal
variation. American Journal of Botany, 71: 759-774.
Hossain, M. A.; Begum, S.; Miah, M. A. S. Uddin, M. J. and Miah, A. J. 1996. Plant
Tissue Culture. Oxford and IBH Publishing Co. Pvt. Ltd. New Delhi, pp. 76-79.
Karim, M. Z.; Amin, M. N.; Hossain, M. A.; Islam, S.; Hossain, F. and Alam, R. 2002.
Micropropagation of two sugarcane (Saccharum officinarum) varieties from callus
culture. On line J.Biological Sci., 2(10): 682-685.
Kochhar, S. L. 1998. Economic Botany in the tropics. Second Edition Macmillan. India
Ltd., p. 476.
Mahmud, K.; Nasiruddin, K.M.; Hossain, M.A. and Hossain, L., 2013. In vitro regeneration
in sugarcane variety Isd 40. Bangladesh J. Sugarcane, 33 & 34: 95-103.
Gopitha, K.; Bhavani, A. L. and Santhilmanickam J. 2010. Effects of the different auxins
and citokinins in callus induction, shoot, root regeneration in sugarcane.
International Journal of Pharma and Bio Science, 1(3): 1-7.
Lal, N. and Singh H. N. 1994. Rapid clonal multiplication of sugarcane through tissue
culture. Plant Tissue Cult., 4(1): 1-7.
Taylor, P. W.J. 1997. Micropropagation of sugarcane (Saccharum spp. Hybrid).
Biotechnology in Agriculture and Forestry, 39: pp. 256-268.

Bangladesh J. Sugarcane, 35 : 79-87 June, 2014

Site Specific Fertilizer Requirement of Sugarcane and
Potato as Intercrop under Sugarcane Based Cropping

System

S. Islam1*, M.A. Haque1, M.S. Islam1 , M.A. Razzak2, M.J. Islam3, S.M.R. Karim4 and
K.M. Alam1
1Soils and Nutrition Division, 2Agronomy and Farming Systems Division
3Physiology and Sugar Chemistry Division, 4Training and Technology Transfer Division
Bangladesh Sugarcrop Research Institute, Ishurdi-6620, Pabna, Bangladesh

ABSTRACT
The field study was conducted to determine the optimum and
economic doses of fertilizers for sugarcane and potato as intercrop at three
different locations of Jamalpur (AEZ 8), Sara, Ishurdi (AEZ 11) and Joypurhat
(AEZ 3) during cropping year 2011-12 and 2012-13. It revealed that sugarcane
yield was increased when it was intercropped with potato for the application of
additional doses of fertilizer, cultural management and its residual effects of
applied fertilizer to the intercrop. Yield data on sugarcane and potato at AEZ 8,
AEZ 11, and AEZ 3, it showed that the highest yield (of two years) of cane of
128.50, 124.00 and 127.00 tha-1 and potato of 12.18, 9.53, 15.82 tha-1,
respectively were obtained from the T6 treatment. Consequently, from the
economic analyses it indicated that the highest economic benefit was also
found from T6 treatment, where soil test based of 100% NPKSZn with 25%
additional dose of NPK for sugarcane (AEZ 8- N226 P29 K216 S18 Zn5, AEZ 11-
N214 P56 K113 S27 Zn3.5 and AEZ 3-N224 P27 K159 S20 Zn3.5) and potato (AEZ 8-N92
P8 K81 S4 Zn2, AEZ 11- N78 P21 K38 S6 Zn1.5 and AEZ 3-N102 P7.5 K60 S5 Zn2) were
applied. Considering the yield and soil fertility status it revealed that the T6
treatment was the best combination for higher sugarcane and potato yield.
Key words: Site specific, intercrop, brix %, yield

INTRODUCTION
Advantage in crop yield from intercropping system is achieved compared to sole
cane due to synergistic effects of companion crops on each other by making better use of
agronomic management and residual advantage of applied fertilizer for intercrops.
Sugarcane crop depletes a considerable amount of nutrients from the soil. As a
consequence soils lose its inherent ability to supply nutrient for sustainable production.
Yadav et al. (1987) reported that the organic matter content of sugarcane soil increased
due to companion crops. The combined return of cane and potato was always higher
when both the crops were planted together and received their recommended doses of
fertilizer (Bokhtiar et al., 2002). Intercropping in sugarcane with potato, garlic, onion and
vegetable is a common practice in cane growing countries. Intercropping has been
recognized as a potential system for augmenting the productivity over space and time in
*Corresponding author: S. Islam, Scientific Officer
e-mail: [email protected]

80 Bangladesh J. Sugarcane, 35 : 79-87 June, 2014

subsistence farming situations. It increases total yield, higher monetary return, greater
resource utilization and fulfil the diversified need of the farmers. Sing et al. (1986) and
Rathi et al. (1974) observed that all combination of mastard potato, onion, fodder and
sugarbeet with sugarcane proved more profitable than growing autum planted cane only.
Rahman et al. (1994) conducted an experiment with mustard, lentil, potato, onion,
tomato, garlic, chickpea and coriander as intercrop with sugarcane and observed that
economic performance of all intercrops shows much higher benefit than sugarcane sole
cropping. Sugarcane is a long duration and widely spaced crop and up to 120 days the
canopy does not cover the vacant space in between the rows, as such there is ample
scope to grow short- duration intercrops. Due to complementary effect of different crops
when grown together, making better use of resources ultimately helps in productivity of
sugarcane. The production of intercrop in sugarcane in Bangladesh is localized
depending on the soil, environment and market. However, information regarding proper
fertilizer management with potato, onion and garlic with sugarcane in intercropping
system is scanty. Hence, the study was undertaken to find out optimum and economic
doses of fertilizers for sugarcane and potato as intercrop under sugarcane based farming
system.

MATERIALS AND METHODS

The experiment was conducted in 2011-12 and 2012-13 cropping seasons in
three locations viz. Jamalpur (AEZ 8), Sara, Ishurdi (AEZ 11) and Joypurhat (AEZ 3) with
eight treatments viz.,T1 = STB 100% NPKSZn, T2 = T1 + 25% N, T3 = T1 + 25% NP, T4 =
T1 + 25% NK, T5 = T1 + 25% PK, T6 = T1 + 25% NPK, T7 = 75% of T1, T8 = Control with
three replications. The experiment was laid out following Randomized Complete Block
Design (RCBD). The plot size was 8m × 6m. The sugarcane variety was Isd 39 and;
Potato variety was Cardinal. From the experimental plot, soil samples collection, sample
processing (drying, sieving, storing) and follow up chemical analysis were done for
determining the fertilizer doses. Nine soil samples at 0-15 cm depths were collected from
each experimental plot. The experiment was set up in the month of November planting of
potato and three eyed bud of sugarcane. One-third of nitrogen and potassium and the
entire quantity of phosphorus, sulphur and zinc were applied in trenches and thoroughly
mixed with the soil prior to planting of sugarcane and potato. One-third potassium and
nitrogen were applied as top dressing at tillering stage (130 days). The remaining quantity
of potassium and nitrogen were top dressed at late-tillering stage (about 180 days).
Intercrop potato was harvested in March and sugarcane in December. Millable cane,
yield of cane and Brix % were recorded from sugarcane, and in respect of potato only the
yield data ware taken.

Yield and yield attributes RESULTS AND DISCUSSION

At Jamalpur site, location 1, from table 1, it was found that in 2011-12 cropping
season, the highest sugarcane and potato yield of 131.0 tha-1 and 14.0 tha-1 respectively
were obtained from T6 treatment which received 25% more NPK than recommended
dose. In 2012-13 cropping season, treatment pTo6thaatovin(1g2T.41+t2h5a%-1).NRPeKsualltsofropmrodthuecesdtuthdey
hieghest yield of sugarcane (118 tha-1) and
indicated that raising of intercrops influenced the sugarcane yield. This result supported
by Paul et al. (2008) where they reported that the potato intercropping increased the

Site Specific Fertilizer Requirement of Sugarcane and Potato as ...... System 81

sugarcane yield. Application of soil test based of 100% NPKSZn with 25% surplus NPK
produded better yield for sugarcane and potato, it might be due to complementary effect
of growing intercrops and residual effect of plant nutrients on sugarcane crop that
ultimately enhanced more cane yield compared to sole cane.
In 2011-2012 cropping season, at Jamalpur site location 2 data on the highest
millable cane (121 × 103 ha-1), yield (135 tha-1), Brix (19.9%) and patato yield (10.8 tha-1)
were found in T6 treatment. In 2012-2013 cropping season recorded data was different
only in respect of millable cane. The highest millable cane (117× 103 ha-1) was found in T3
treatment which was received soil test based of 100% NPKSZn with 25% surplus NP but
the highest yield of sugarcane (130 tha-1) and potato (12.4 tha-1) was found in T6
treatment which was received 25% more NPK than recommended dose. It is indicated
that intercropping with sugarcane had no adverse effect on sugarcane yield. Moreover
residual effect of added fertiltizer and cultural management for the intercrops resulted
more sugarcane yield. Adhikari and Karki (2006) tested potato experiment for three years
in Agricultural Research Station(ARS), Napal and observed that increasing rate of
potassium (0-100 kg ha-1) significantly increased tuber yield (24.75 t ha-1) Table 2.

At Ishurdi site location Sara
Data on millable cane (97×103 ha-1), sugarcane yield (111 tha-1) and potato yield
(9.3 tha-1) was higher in T6 treatment in 2011-12 but in respect of millalble cane potato
yield there were no significant different among the treatment T1, T2, T3, T4, and T5
treatments. It seemed that combination of soil test based of 100% NPKSZn different
packages miantained soil fertility status for different growing season. In 2012-2013
cropping season, the highest millable cane (108 ×103 ha-1), cane yield (137 tha-1) and potao
yield (9.8 tha-1) was found from T6 treatment where soil test based of 100% NPKSZn with
25% surplus NPK was applied. Soomro et al., (2014) observed that tillers plant-1, stem
girth, internodes plant-1, millable canes, and dry matter were higher with the application of
25% more than recommended fertilizer at 281.25-140-210 NPK kg ha-1 Table 3.

At Joypurhat site
In 2011-2012 cropping season, data on millable cane (117×103 ha-1), cane yield
(115 tha-1) and potato yield (17.0 tha-1) was the higher in T6 treatment. In 2012-2013
NcrPoKppwinagssreeacsoordne, dT6htereigahttmmenilltahbalevincagnseoi(l9t8e×s1t 0b3ashead-1)o,fc1a0n0e%yiNelPdK(1S3Z9n with 25% surplus
tha-1) and potato
yield (14.6 tha-1). This result supported by Dashora (2012) and reported that increasing
rate of NPK up to 200:60:40 kg/ha significantly increased cane yield Table 4.

Economics of fertilizer use
The economic analysis of different fertilizer packages was done considering the

total cost and gross return of the treatments. The economic benefit was greatly varied at
different sites. At Jamalpur site location 1, in sugarcane-potato intercroping system the
BCR was found 6.24. At Jamalpur site location 2, the highest economic benefit of 7.76
was found in T6 which was followed by T5 of 7.21. At Ishurdi site location Sara, the
highest economic benefit of 6.50 was obtained from T6 treatment which was closely
related to T5 of 5.47. At Joypurhat site, the highest economic benefit of 7.42 was found in
T6 which was followed by T5 of 6.88 (Table 5-8).

82 Bangladesh J. Sugarcane, 35 : 79-87 June, 2014

Therefore, from the economic analyses of four locations it was found that the
highest MBCR/economic benefit was found from T6 treatment, where soil test based of
100% NPKSZn with 25% additional dose of NPK for sugarcane (AEZ 8- N226
P29K216S18Zn5, AEZ 11- N214 P56K113S27Zn3.5 and AEZ 3-N224 P27K159S20Zn3.5) and potato
(AEZ 8-N92P8K81S4Zn2, AEZ 11- N78P21K38S6Zn1.5 and AEZ 3-N102P7.5K60S5Zn2) were
applied. Similarly at AEZ 8, AEZ 11, and AEZ 3 the highest yield of cane of 128.40,
124.15 and 127.00 tha-1 and potato of 12.18, 9.53, 15.82 tha-1 , respectively were also
obtained from the T6 treatment. Considering the yield and soil fertility status it revealed
that the T6 treatment was the best combination for higher sugarcane and potato yield.
Soil fertility status

The initial and post harvest nutrient status of soil pH, organic carbon, total N,
available P, K, S and Zn for all sites are presented in Table 9-12. There are considerable
decrease in organic matter in soil of Jamalpur site location 1, Ishurdi site location Sara
and Joypurhat site but organic matter content of Jamalpur site location 2 was more or
less same. The changes of total N, available P, K, S and Zn were not conspicous due to
single year practice of using different ertilizer doses in experimental plots under study.

Table 1. Performances of sugarcane with potato as intercrop in different nutrient
management packages at Jamalpur (location 1, AEZ 8)

2011-12 2012-13
Sugarcane
Sugarcane Potato Potato
Yield Yield
Treatment Millable Yield Brix (%) (tha-1) Millable cane Yield Brix (%) (tha-1)
cane (tha-1) (× 103 ha-1) (tha-1)
T1= STB 100% 11.5abc 9.9 bc
NPKSZn (× 103 ha-1)
T2 = T1 + 25% N 11.2bc 9.9 bc
117a 123ab 17.0c 11.6abc 79bc 100bc 17.8abc 10.8 b
12.1ab 10.4 b
115a 121ab 17.2bc 12.9ab 67cd 94cd 18.6 a 11.7 a
94ab 112a 16.9cd 12.4 a
T3 = T1 + 25% NP 119a 123ab 16.7c 14.0a 93ab 109ab 18.5ab 9.4 c
9.7c 96a 98cd 17.4bcd 6.1 d
T4 = T1 + 25% NK 119a 125a 18.2abc 6.6d 93.3ab 118a 17.9abc 0.9049
2.12 66.9cd 89de 16.6d
T5 = T1 + 25% PK 119a 125a 17.0c 56.6d 80e 17.3cd
15.36 9.713 1.036
T6 = T1 + 25% NPK 114a 131a 19.7a

T7 = 75% of T1 113a 108b 18.9ab

T8 = Control 91b 64c 17.8bc

LSD (0.05) 15.66 15.11 1.53

T1 = N 180P38 K170 S20 Zn5.6 (sugarcane) and N73 P11 K65 S4 Zn2 (potato) for 2011-12
T1 = N190 P18 K180 S18 Zn5.6 (sugarcane) and N78 P5 K65 S6 Zn1 (potato) for 2012-13

Site Specific Fertilizer Requirement of Sugarcane and Potato as ...... System 83

Table 2. Performances of sugarcane with potato as intercrop in different nutrient
management packages at Jamalpur site (location 2, AEZ 8)

2011-12 2012-13
Sugarcane
Sugarcane Potato Potato

Treatment Millable Yield Brix (%) Yield Millable cane Yield Brix (%) Yield
cane (tha-1) (tha-1) (× 103 ha-1) (tha-1) (tha-1)
T1= STB 100% NPKSZn
T2 = T1 + 25% N (× 103 ha-1) 7.2c 9.9b
T3 = T1 + 25% NP 9.5b 10.6b
T4 = T1 + 25% NK 116a 121b 17.8bcd 7.0c 108ab 103b 17.2bcd 10.4b
T5 = T1 + 25% PK 7.5c 111a 112b 17.0cde 11.0b
T6 = T1 + 25% NPK 112a 116b 17.6cd 9.0b 117a 109b 17.9ab 11.0b
T7 = 75% of T1 10.8a 108ab 114b 12.4a
T8= Control 110a 115b 17.3d 7.0c 104ab 124a 16.2e 8.3c
LSD (0.05) 5.2d 108ab 130a 18.0ab 6.0d
115a 121b 18.4bc 1.00 102ab 104b 17.7bc 0.979
93b 60c 18.7a
119a 125ab 18.3bc 16.8de

121a 135a 19.9a

114a 102c 18.6b

88b 62d 17.8bcd

9.912 10.14 0.76 14.19 9.98 0.812

T1= N183 P18 K170 S20 Zn4.5 (sugarcane) and N75P5K65S4Zn1.8 (potato) for 2011-12
T1= N171P18K174 S15Zn4.5 (sugarcane) and N70 P5 K65 S3 Zn2 (potato) for 2012-13

Table 3. Performances of sugarcane with potato as intercrop in different nutrient
management packages at Ishurdi site (location Sara, AEZ 11)

2011-12 2012-13

Treatment Sugarcane Potato Sugarcane Potato
Yield
T1= STB 100% Millable Yield Brix (%) (tha-1) Millable Yield Brix Yield
NPKSZn cane (tha-1) 8.6 ab cane (tha-1) (%) (tha-1)
T2 = T1 + 25% N
T3 = T1 + 25% NP (× 103 ha-1) (× 103 ha-1) 8.6 c
T4 = T1 + 25% NK
T5 = T1 + 25% PK 91 a 99 ab 19.2 95 b 110 b 18.2 8.7 c
T6 = T1 + 25% NPK 8.2 d
T7 = 75% of T1 89 a 90 b 18.3 8.4 ab 95 b 114 b 18.1 8.6 c
T8= Control 91 a 88 b 9.2 b
LSD (0.05) 89 a 89 b 18.7 7.8 abc 92 b 109 b 18.6 9.8 a
92 a 97 ab 7.0 e
97 a 111 a 18.5 8.6 ab 98 b 108 b 17.9 5.7 f
77 b 82 b 0.19
73 b 64 c 18.7 9.0 ab 98 b 115 b 18.4
8.34 16.22
18.6 9.3 a 108 a 137 a 17.9

19.0 7.2 bc 95 b 96 c 18.3

18.9 6.2 c 53 c 55 d 18.5

NS 1.81 8.63 11.63 NS

T1= N183 P18 K170 S20 Zn4.5 (sugarcane) and N75P5K65S4Zn1.8 (potato) for 2011-12
T1= N171P18K174 S15Zn4.5 (sugarcane) and N70 P5 K65 S3 Zn2 (potato) for 2012-13

84 Bangladesh J. Sugarcane, 35 : 79-87 June, 2014

Table 4. Performances of sugarcane with potato as intercrop in different nutrient
management packages at Joypurhat site (AEZ 3)

2011-12 2012-13

Treatment Sugarcane Potato Sugarcane Potato

T1 = STB 100% PKSZn Millable Yield Brix Yield Millable Yield Brix Yield
T2 = T1 + 25% N cane (tha-1) (%) (tha-1) cane (tha-1) (%) (tha-1)
T3 = T1 + 25% NP
T4 = T1 + 25% NK (× 103 ha-1) 15.6 a (× 103 ha-1) 13.4 ab
T5 = T1 + 25% PK 14.5 a 13.3 ab
T6 = T1 + 25% NPK 115 a 105 bcd 18.3 16.7 a 87 b 112 c 17.5 13.4 ab
T7 = 75% of T1 16.8 a 12.9 b
T8 = Control 93 b 97 d 19.2 15.8 a 91 ab 123 b 16.5 13.1 b
LSD (.05) 17.0 a 14.6 a
93 b 100 cd 19.0 15.2 a 90 ab 122 b 16.6 10.5 c
11.1 b
105 ab 109 abc 18.8 2.51 87 b 112 c 16.3 8.9 d
1.22
109 ab 110 ab 18.6 91 ab 127 b 16.6

117 a 115 a 18.8 98 a 139 a 16.5

105 ab 86 e 19.3 83 b 102 d 16.8

91 b 66 f 18.6 71 c 68 e 16.9

17.78 8.21 NS 8.06 9.17 NS

T1= N171P25 K127 S16 Zn3.5 (sugarcane) and N85 P7 K48 S6 Zn1.7 (potato) for 2011-12
T1= N188 P18 K127 S24 Zn3.5 (sugarcane) and N78 P5 K48 S4 Zn2 (potato) for 2012-13

Table 5. Economic analysis of different treatment of Jamalpur site (location 1,
AEZ 8)

Treatment Yield (tha-1) Adjusted Cost of fertilizer Total Gross Additional
Cane Potato cane yield (Tk.ha-1) cost of Return profit MBCR
T1 fertilizer (Tk.ha-1) (Tk.ha-1)
T2 (tha-1) Cane Potato ((TTkk..hhaa--11))
T3 28359.22
T4 111.49 10.7 160.87 18582.04 9777.18 31614.12 418262 153686 5.42
T5 107.68 10.55 156.37 20600.14 11013.98 406562 154986 4.49
T6 117.88 11.18 169.48 21425.14 11343.98 32769.12 440648 176072 5.37
T7 117.3 11.25 169.22 21905.14 11733.98 33639.12 439972 175396 5.21
T8 12.29 168.72 20712.04 10827.18 31539.22 438672 174096 5.52
112 13.21 185.27 22730.14 12063.98 481702 217126 6.24
124.3 9.55 143.03 13982.02 34794.12 371878 107302 4.87
98.95 6.33 101.76 8036.6 22018.62 264576
72.54 - - - -
-

TPTk1K=.1;S5TTk6Bg=-11T,01G0+%yp2sN5u%PmKNS: PTZKkn.1;; 0TT72kg==-17T,51Z%+in2co5fs%Tu1lpN;hTa;8tTe=3:=CToTkn.11t+r5o02l.5kU%gr-e1N,aPP:rTi;ckTe.4o2=f0sTku1gg+-a1,2rcT5aS%nPeN::KTTkk;..T22562=0k0Tgt1-1-1,+,M2o5P%:
potato : 12000 t-1.

Site Specific Fertilizer Requirement of Sugarcane and Potato as ...... System 85

Table 6. Economic analysis of different treatment of Jamalpur site (location 2,
AEZ 8)

Treatment Yield (tha-1) Adjusted Cost of fertilizer Total Gross Additional MBCR
cane (Tk.ha-1) fertilizer Return profit
T1 yield cost (Tk.ha-1) (Tk.ha-1) 6.02
T2 Cane Potato (tha-1) Cane Potato (Tk.ha-1) 6.28
T3 5.48
T4 112.04 8.25 150.09 17674.53 9803.63 27478.15 390244 165392 5.84
T5 113.94 10.06 416891 192039 7.21
T6 112.28 8.72 160.34 19582.93 10976.63 30559.55 396555 171703 7.76
T7 117.05 9.26 415377 190525 6.50
T8 124.32 152.52 20132.93 11196.63 31329.55 443232 218380
132.5 10 483640 258788 -
103.04 11.6 159.76 20872.93 11766.63 32639.55 359931 135079
60.52 7.62 224852
5.63 170.47 19511.53 10759.63 30271.15 -

186.02 21422.93 11931.63 33354.55

138.44 13395.11 7396.34 20791.45

86.48 - - -

T1= STB 100% NPKSZn ; T2 = T1 + 25% N ; T3 = T1 + 25% NP ; T4 = T1 + 25% NK ; T5 = T1 + 25%
PK ; T6 = T1 + 25% NPK ; T7 = 75% of T1 ; T8= Control

Table 7. Economic analysis of different treatment of Ishurdi site location Sara
(AEZ 11)

Treatment Yield (tha-1) Adjusted Cost of fertilizer Total Gross Additional
cane (Tk.ha-1) variable Return profit MBCR
T1 yield (Tk.ha-1) (Tk.ha-1)
T2 Cane Potato (tha-1) Cane Potato cost
T3 141.19 (Tk.ha-1)
T4 104.5 8.6 17892.39 9174.27 27066.66 367094 143494 5.3
T5 101.9 8.54 141.32
T6 98.7 135.62 19736.59 10220.07 29956.66 367432 143832 4.8
T7 98.4 8 138.23 21001.59 10811.57 31813.16 352612 129012 4.06
T8 106.05 8.63 20426.59 10551.07 30977.66 359398 135798 4.38
124.15 9.31 149.02 19847.39 10095.27 387452 163852 5.47
88.9 9.53 168.13 21691.59 11136.57 29942.66 437138 213536 6.5
59.11 7.1 13630.17 6999.13 32828.16 316342 92742 4.5
5.96 121.67 223600
86 - - 20629.3 - -
-

T1= STB 100% NPKSZn ; T2 = T1 + 25% N ; T3 = T1 + 25% NP ; T4 = T1 + 25% NK ; T5 = T1 + 25%
PK ; T6 = T1 + 25% NPK ; T7 = 75% of T1 ; T8= Control

Table 8. Economic analysis of different treatment of Joypurhat site (AEZ 3)

Treatment Yield (tha-1) Adjusted Cost of fertilizer Total Gross Additional
cane yield (Tk.ha-1) variable Return profit MBCR
T1 (Tk.ha-1) (Tk.ha-1)
T2 Cane Potato (tha-1) Cane Potato cost
T3 (Tkha-1)
T4 108.45 14.52 175.46 15800.82 9028.24 24829.06 456196 162006 6.52
T5 110 13.93 174.39 17748.22 10313.84 453414 159224 5.67
T6 15.05 180.51 18366.72 10575.34 28062.06 469326 175136 6.05
T7 111.05 14.85 179.04 18704.22 10853.84 28942.06 462904 168714 5.75
T8 110.5 14.45 185.14 17375.32 9829.74 29558.06 481364 187174 6.88
118.45 15.82 19322.72 11115.34 520000 225810 7.42
127 12.84 200 11874.24 6894.38 27205.06 398476 104286 5.56
153.26 30438.06 294190
94 10 113.15 - - - -
67 18768.62
-

T1= STB 100% NPKSZn ; T2 = T1 + 25% N ; T3 = T1 + 25% NP ; T4 = T1 + 25% NK ; T5 = T1 + 25%
PK ; T6 = T1 + 25% NPK ; T7 = 75% of T1 ; T8= Control

86 Bangladesh J. Sugarcane, 35 : 79-87 June, 2014

Table 9. Nutrient status of initial and post harvest soil at Jamalpur site (location 1,
AEZ 8)

Treatment pH OC (%) (N %) P K (meq%) S (mg kg-1) Zn
(mg kg-1) (mg kg-1)
Initial soil
6.12 0.33 0.065 14 0.10 20 0.41
T1= STB 100% NPKSZn
T2 = T1 + 25% N Post harvest soil 0.40
T3 = T1 + 25% NP 0.41
T4 = T1 + 25% NK 6.20 0.35 0.070 15 0.12 19 0.39
T5 = T1 + 25% PK 0.38
T6 = T1 + 25% NPK 6.15 0.33 0.070 14 0.11 18 0.39
T7 = 75% of T1 0.40
T8 = Control 6.12 0.34 0.065 15 0.12 18 0.39
0.38
6.15 0.35 0.070 15 0.12 19

6.20 0.35 0.065 14 0.13 18

6.20 0.34 0.070 16 0.13 21

6.15 0.33 0.060 14 0.11 19

6.10 0.30 0.060 13 0.10 18

Table 10. Nutrient status of initial and post harvest soil at Jamalpur site (location
2, AEZ 8)

Treatment pH OC (%) N (%) P (mg kg-1) K (meq%) S (mg kg-1) Zn (mg kg-1)
Initial soil 5.95 0.42 0.06 25 0.10 20 0.57
Post harvest soil
T1= STB 100% NPKSZn 6.00 0.40 0.06 22 0.11 18 0.60
T2 = T1 + 25% N 6.10 0.41 0.07 21 0.12 19 0.58
T3 = T1 + 25% NP 6.05 0.39 0.12 18 0.58
T4 = T1 + 25% NK 6.10 0.39 0.07 22 0.13 19 0.59
T5 = T1 + 25% PK 5.95 0.40 0.12 19 0.58
T6 = T1 + 25% NPK 6.10 0.41 0.07 21 0.12 20 0.58
T7 = 75% of T1 6.00 0.40 0.11 18 0.57
T8= Control 5.98 0.38 0.07 22 0.09 18 0.55

0.07 23

0.06 22

0.05 21

Table 11. Nutrient status of initial and post harvest soil at Ishurdi site (location
Sara, AEZ 11)

Treatment pH OC (%) N (%) P (mg kg-1) K (meq%) S (mg kg-1) Zn
Initial soil 7.50 (mg kg-1)
0.58 0.070 7.0 0.20 31
0.72
Post harvest soil 0.19 25
0.20 26 0.70
T1= STB 100% NPKSZn 7.55 0.55 0.065 8.0 0.21 25 0.68
T2 = T1 + 25% N 7.60 0.20 26 0.68
0.54 0.065 7.5 0.20 25 0.65
T3 = T1 + 25% NP 7.60 0.21 26 0.66
T4 = T1 + 25% NK 7.58 0.55 0.060 7.5 0.19 25 0.68
T5 = T1 + 25% PK 7.6 0.18 24 0.65
T6 = T1 + 25% NPK 7.65 0.58 0.060 7.0 0.65
T7 = 75% of T1 7.60 0.56 0.065 7.5
0.57 0.070 8.0
T8= Control 7.50
0.56 0.065 7.0

0.55 0.055 6.5

Site Specific Fertilizer Requirement of Sugarcane and Potato as ...... System 87

Table 12. Nutrient status of initial and post harvest soil at Joypurhat site (AEZ 3)

Treatment pH OC (%) N (%) P (mg kg-1) K (meq%) S (mg kg-1) Zn
Initial soil 6.20 0.17 22 (mg kg-1)
0.42 0.080 20.0
0.74
0.40 Post harvest soil
0.41 0.65
T1 = STB 100% NPKSZn 6.25 0.40 0.070 19.0 0.16 20 0.6
T2 = T1 + 25% N 6.22 0.41 0.070 19.0 0.65
T3 = T1 + 25% NP 6.20 0.41 0.065 19.5 0.16 19 0.67
T4 = T1 + 25% NK 6.18 0.42 0.070 19.0 0.68
T5 = T1 + 25% PK 6.20 0.40 0.15 20 0.70
T6 = T1 + 25% NPK 6.25 0.40 0.68
T7 = 75% of T1 6.18 0.17 20 0.65
T8 = Control 6.15
0.070 19.5 0.18 19
0.075 19.5
0.060 19.0 0.18 20

0.16 19

0.065 18.0 0.15 17

REFERENCES
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potato plants to potassium fertilizer rates and soil moisture deficit. Adv. Appl. Sci.
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sugarcane-based croping systems. Pak.J.Biol. Sci., 5:165-168.
Bokhtiar, S.M.; Hossain, M.S.; Mahmud, K. and Paul, G.C. 2003. Site specific nutritient
management for sugarcane-potato and sugarcane-onion in intercropping
systems. Asian Journal of Plant Sciences, 2:1205-1208.
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genotypes under various planting seasons and fertility levels in South-East
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fertilizer requirement of sugarcane and intercrop (potato) under sugarcane based
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Rathi, K.S.; Tripathi, H.N. and Singh, D. 1974. Studies on intercropping rabi crop in
autum planted sugarcane. Indian Sug. J., 24: 201-205.
Singh, V.S.; Kothari, K. and Tripathi, H.N. 1986. Studies on intercropping in sugaracane
in central uttar pradesh. Indian Sug. J., 35: 559-562.
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of inorganic NPK fertilizers under different proportions on growth, yield and juice
quality of sugarcane (Saccharum officinarum L). Pure Appl. Bio., 3(1), 10-18.
Tahman, M.S.; Haq, M.F.; Islam, M.S.; Bashar, M.K.; Ara, N. and Sardar, P.K. 1994.
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32: 80-84.

Bangladesh J. Sugarcane, 35 : 88-95 June, 2014

Effects of Phyto Clarificant as Powder Form on Goor
Manufacture

M.S. Arefin1*, M.K. Begum1, M.J. Islam1, M.A. Rahman2, A.K. Ghose3 and S. Islam4
1 Physiology and Sugar Chemistry Division, 2 On-farm Research Division
3 Biotechnology Division, 4 Soils and Nutrition Division

Bangladesh Sugarcrop Research Institute, Ishurdi-6620, Pabna, Bangladesh

ABSTRACT
In goor manufacturing sodium hydrosulphite (hydrose) is
indiscriminately used for cane juice clarification beyond recommended limits
(35 g hydrose/1000 L juice) to impart light golden yellow colour to goor. Often
level of SO2 in goor exceeds beyond 70 ppm which is not suitable for human
consumption. It is reported that prolonged intake of hydrose added goor can
create bowel irritation, chronic dysentery and gastro urinal troubles. Many
phyto clarificants have been studied and extracts of wild okra use. Amongst
the phyto clarificants found to be quite effective juice clarificants for goor
making. Since Wild Okra plant is not available during peak period of goor
manufacturing, so technique has been developed to convert effective phyto
clarificant as powder form for availability throughout the year for goor
processing. An experiment was conducted in Physiology and Sugar Chemistry
Division of the Bangladesh Sugarcane Research Institute (BSRI), Ishurdi,
Pabna during 2011-2012 and 2012-2013 crushing year to determine the dose
of phyto clarificants as powder form for preparation of goor through cane juice
clarification. The experiment comprised of five treatment combinations
following Complete Block (CBD) Design with three replications. Goor were
prepared by simple open pan boiling method using 0 g, 20 g, 30 g, 40 g and 50
g of Wild Okra powder per 100 liter cane juice at different months (Oct, Dec
and Feb). Superior quality of goor associated with higher sucrose content,
higher colour transmittance, lower reducing sugars content, low moisture
content and high goor recovery. The presence of this parameter in right
proportion in juice is essential for producing good quality goor with standard
keeping quality. On the basis of physical and chemical properties of goor, it
was concluded that the treatment T2 (30 g Wild Okra powder per 100 L cane
juice) was used for better juice clarification and produce lucrative colour and
better quality goor.
Key words: Sugarcane, phyto clarificant, powder, goor manufacture

INTRODUCTION
Sugarcane (Saccharum officinarum) is the main source of raw material for
production of goor in Bangladesh (Arefin et al., 2011). Goor is manufactured almost all
parts of Bangladesh where sugarcane is grown in abundance. Selection of sugarcane
varieties/clones suitable for quality goor production is important. The varieties suitable for

* Corresponding author: M.S. Arefin, Senior Scientific Officer
e-mail: [email protected]

Effects of Phyto Clarificant as Powder Form on Goor Manufacture 89

goor making are found to be equally good for sugar production. However, a variety
suitable for sugar production may not be equally suitable for goor making (Singh et. al.,
1975). Goor quality is considered by its sucrose content, reducing sugars content, colour
etc. (Gopal and Chiranjivi, 1959). Though the quality may be further modified by method
of boiling and clarificants used for preparation of goor. Chemical composition of goor and
its quality are dependent on composition of juice from which goor is made. Goor is high
calorie sweetener and as it contains minerals, protein, glucose and fructose, it is known
to be healthier in comparison to white sugar. A good quality goor contains more than 70%
sucrose, less than 10% of glucose and fructose, less than 5% minerals and less than 3%
moisture (Nath et al., 2015). Goor production involves extraction of juice and its
clarification. The clarification of juice depends on the composition of juice that affects the
quality of goor. Besides sugars, it contains suspended impurities in the form of coarse
particles and colloids. Soil particles, wax, fat, protein, gum, pectin, tannins, and coloring
matters are extracted from the cane during juice extraction and they remain in colloid
form (Rao, 1984).

In contrast, the plantation white sugar contains only 99.5% sucrose without any
minerals. Commercial sugar has been implicated as a causative factor in certain heart
diseases and also causing as a primary factor in dental problems. Therefore, the
nutritional potential of goor is considerable value for majority of the population living in
rural Bangladesh. Compared to goor the white sugar takes away calcium and potassium
from the body during digestion apart from difficulty in digestion of white sugar. A quality
goor is one having golden yellow colour, hard in texture, crystalline in nature, sweet in
taste, less in impurities and low in moisture. The quality of goor is influenced by the
variety of cane grown, quantity of fertilizers used and quality of irrigation water and the
method of processing adopted. The common man is often misguided by the impression
that the brightly coloured goor is the best quality goor. This is mostly wrong since in the
present day for obtaining better colour most of the goor is produced by the use of harmful
chemical clarificants. Many a times in order to get solidification of goor from bad quality
juice and to get bright colour the goor making artisans add several unrecompensed
chemicals and substances.

In the present day goor is prepared mostly from sugarcane grown by using
chemical fertilizers, herbicides and pesticides and also preparation of goor involves
mostly chemical clarificants. The quality of goor prepared with the commonly used
chemical clarificants such as hydrose, sodium carbonate, sodium bi-carbonate, sajji,
super phosphate, alum etc., not only has temporary improvement in colour, salty taste
and poor storability but also excess use of them may result in harmful residues such as
sulphur dioxide beyond prescribed limit. Many times the market goor has been found to
contain excess quantities of harmful chemicals like sulphur dioxide. Due to use of such
chemicals the taste and storability of such goor is also affected. An analysis results of
available goor in the market has revealed that most of the goor are prepared by the use
of chemicals which contain more than 80-120 ppm of sulphur di-oxide, which are well
above the prescribed norms of 50 ppm by Indian standard (IS 12923): 1990. This high
amount of sulphur di-oxide is detrimental to the beneficial intestinal microflora leading to
digestive disorders and gastrointestinal problems etc., and also can cause breathing
problems in asthmatic patients. It can also cause colon/rectal cancer and can also
destroy the formation of vitamin A and vitamin B1.

90 Bangladesh J. Sugarcane, 35 : 88-95 June, 2014

In this context, growing of sugarcane only with organics without chemical
fertilizers, weedicides and pesticides and also preparation of goor with use of organic
clarificants assumes importance in order to produce quality goor. There is a growing
demand for organically produced goor both within the country and in the export market. In
general term clarification means the extraction or separation of desired material and
discarding the rest in a particular system either by means of chemical treatment or by
mechanical operation. At times both may be applied for ultimate degree of separation
requirement. In the instant proposition the cane juice which is a colloidal suspension of
inorganic and organic non-sugars along with dissolved impurities needs dual operation
followed by thickening, crystallization and centrifugation for the manufacture of sugar.
Clarification of sugarcane juice can be classified into two categories such as chemical
clarification and mechanical clarification. Commonly vegetative clarificants are used in
boiling pan. Some chemicals such as hydrose (sodium hydro-sulphite), lime (calcium
oxide), sodium bicarbonate, sodium carbonate, super phosphate and alum have been
used in combination with the vegetative clarificants. A good flocculant should increase the
settling rate of insoluble solids, decrease the mud volume, produce good clarity of
clarified juice with the least turbidity and should produce good filterability of mud, with
good clarity of filtrate (Panda et al., 2008).

The synthetic clarificant like Bhendi powder SNi @ 2 ppm with herbal clarificant
bhendi plant @ 2 kg/1000 liter were found effective in improving non reducing sugar
(NRS), Colour, goor recovery and maximum removal of scum, showing better effect on
quality of goor and also helped in maintaining higher NRS and better colour goor during
storage (Patil et al., 2005). In goor manufacturing sodium hydrosulphite (hydrose) is
indiscriminately used for cane juice clarification beyond recommended limits (35 g
hydrose/1000 L juice) to impart light golden yellow colour of goor. Often level of SO2 in
goor exceeds beyond 70 ppm which is not suitable for human consumption (Bureau of
Indian Standard I.S.12923, 1990). It is reported that prolonged intake of hydrose added
goor can create bowel irritation, chronic dysentery and gastro urinal troubles. Chemical
clarificants adversely affect the human health beings since traces of chemicals remain in
the final product (Anjal, Tagare 1972). All these chemicals (except lime) brighten colour of
goor initially, but the colour of the goor becomes dull during storage. So alternative
methods of clarification is being explored to alleviate the use of chemical clarificants
(Vishal, 2003). Many plant clarificants have been reported during the last 60 years.
Amongst the plant clarificants frsesh Wild Okra plant was found to be quite effective juice
clarificant for goor making. Since Wild Okra plant is not available during peak period of
goor manufacturing, so technique has been developed to convert effective phyto
clarificants as powder form for availability throughout the year for goor processing. The
present experiment was undertaken with objectives to determine the dose of phyto
clarificants as powder form for cane juice clarification

MATERIALS AND METHODS

The experiment was conducted in Physiology and Sugar Chemistry Division of
the Bangladesh Sugarcane Research Institute (BSRI), Ishurdi, Pabna during two crusing
seasons 2011-2012 and 2012-2013. The experiment was laid out in complete block
design (CBD) with five treatments combination and replicated three times. Goor was
prepared by simple open pan boiling method using 0 g, 20 g, 30 g, 40 g and 50 g of wild

Effects of Phyto Clarificant as Powder Form on Goor Manufacture 91

okra powder per 100 liter cane juice at different months (Oct, Dec and Feb). The powder
clarificant was prepared from Wild Okra stem collected at seed formation and maturity
growth stages of the plant. Wild Okra stem was thoroughly washed by clean water. After
air drying stem and branches were separated out and then scrapped by using hand sharp
knife. The scrapped sample material (bark) was dried by air under shade for several
days. The shade dried stem was subjected to grinding machine for preparation of
powder. After grinding, powder was sieved out from fibrous stem material. Different doses
0 g, 20 g, 30 g, 40 g and 50 g of Wild Okra powder was suspended in 2 liter clean water
and mixed thoroughly. After 1 hour mucilaginous extract was filtered. Mucilaginous filtrate
was used for clarification of 100 liter of boiling cane juice. The commercial variety was
used as test crop. Goor was prepared with and without powder phyto clarificant. Physical
and chemical properties of prepared goor viz., texture, crystalline nature, colour in solid
state, taste, sucrose%, colour transmittance (at 0.25N solution), pH, reducing sugars per
cent etc. were determined by following method of Roy (1954). Data on sucrose%, colour
transmittance, moisture% and reducing sugars% of prepared goor were recorded.
Recorded data have been presented graphically for discussion.

RESULTS AND DISCUSSION
Physical and chemical properties of goor

The physical and chemical properties of goor had been studied immediately after
its preparation. Goor is normally priced in the market for its colour. The higher colour unit
indicates better is the quality. In case of physical properties of goor, treatment T1 (20 g
Wild Okra powder per 100 L cane juice), T2 (30 g Wild Okra powder per 100 L cane juice),
T3 (40 g Wild Okra powder per 100 L cane juice) and T4 (50 g Wild Okra powder per 100 L
cane juice) showed superior quality goor due to golden colour, good crystalline nature
and taste compared to T0 (control) (Table 1).

The colour transmittance of goor is shown in the Figure 3. It is seen that the
highest colour transmittance of goor was obtained from the treatment T2 and the value is
67.0% followed by T1 (57.0%) and T3 (50.0%). The lowest colour transmittance was
obtained in the treatment T0 (45.0). It is evident from the data that colour transmittance of
goor prepared from the treatment T2 (powder phyto clarificant 30 g/100 L cane juice) was
comparatively highest value. Similar findings were also reported by Sarker, et al. (1985)
that light coloured goor is always preferred by consumers for eating purpose and good
quality goor is characterized by light colour.

The highest sucrose% of goor was recorded from the treatment T2 (75.2%). The
second highest value of sucrose% goor was observed from the treatment T3 (71.29%)
which was statistical similar with the treatment T1 (69.05%). The lowest sucrose% goor
was obtained in the treatment T4 (Figure 1). It is universal that sucrose% being the main
sweetening factor of goor. It was seen from the figoore 2, the lowest reducing sugars% of
goor was found from the treatment T2 (5.65%). The highest reducing sugars% of goor
was observed from the treatment T4 (6.67%) which is statistically similar to the treatment
T3 (6.32%). Results agree with results obtained by Jabber et al. (2005) who reported that
a good quality goor should have higher sucrose% and lower reducing sugars%. Goor
containing higher percentage of reducing sugars are generally hygroscopic, low shelf-life
and ultimately poor quality in nature.

92 Bangladesh J. Sugarcane, 35 : 88-95 June, 2014

From the figoore 4, we observed that there was no significant difference in pH of
prepared goor from all the treatments. Lower pH indicates acidic in nature. Figure 4
shows that the highest pH value of goor prepared from the treatment T2 (5.79) followed
by the treatment T3 (5.61), respectively and prepared goor from these treatments are
suitable for consumption. This result agreed with Arefin et al. (2010). Figure 5
mentioned that there was no significant difference in moisture% of prepared goor from all
the treatments. The highest moisture% and the lowest moisture% obtained from the
treatment T3 (7.91%) and the treatment T2 (7.15%). Higher percentage of moisture
containing goor is not suitable for long term preservation (Gaur et. al., 1979). Figure 6
stated that the highest recovery% of goor was obtained from the treatment T2 (10.15%)
which is statistically similar to the other treatments.

On the basis of above results, it may be stated that superior quality of goor
associated with higher sucrose, high colour transmittance, lower reducing sugars content,
low moisture content and high goor recovery. The correct proportion of this parameter is
essential for producing good quality goor with standard keeping quality. From the
observed physical and chemical properties of goor, it may be concluded that the the
treatment T2 (Powder phyto clarificant 30 g/100 L cane juice) was found better result for
juice clarification and produce lucrative colour and better quality goor (Figure 7).

Table 1. Physical properties of goor under different treatments

Variety/Clone Texture Crystalline Colour in Solid Taste
Nature State
T0 = 0 g WOP/100 L juice Hard Good Sour
T1 = 20 g WOP/100 L juice Hard Good Dark Brown Good
T2 = 30 g WOP/100 L juice Hard Good Golden Good
T3 = 40 g WOP/100 L juice Hard Golden Good
T4 = 50 g WOP/100 L juice Hard Good Good
Good Golden
WOP = Wild Okra Powder Golden