Article

Induction and Evaluation of Somaclonal Variation in

Sugarcane (Saccharum officinarum L.) var.  Isd-16

 

Mohashweta Roy1, Monzur Hossain2, Animesh Biswas1, Rafiul Islam2, Shipra Rani Sarker2

and Sharoni Akhter3  

1Department of Botany, Govt. B. L. College, Khulna, Bangladesh.

2Plant Breeding and Gene Engineering Laboratory, Department of Botany, University of

Rajshahi, Rajshahi-6205, Bangladesh.

3Division of Genome and Biodiversity Research, National Institute of Agrobiological

Sciences (NIAS), Tsukuba Ibaraki, Japan.

 

Received July 23, 2010 Accepted August 23, 2010  

Abstract

Plant regeneration was established through somatic embryogenesis for sugarcane

variety Isd-16 using leaf sheath explants. The explants were cultured onto MS medium

supplemented with different concentrations of 2,4-D for callus induction. The calli were

underwent embryogenesis and produced huge number of somatic embryos when they

were transferred onto MS medium fortified with L-proline. The embryos were

germinated and developed to plantlets on half-strength MS0 and successfully

transplanted into the experimental field and grown to maturity. A large number of

somatic embryos derived plants (SEDPs) were found morphologically different with

some distinct characters such as stool habit, tillering habit, tillering density, auricle,

legule, stalk colour and bud shape as compared to setts derived plants (SETDPs).

Significant differences were also noted between SEDPs and SETDPs in respect of stalk

height, tillers/plant, five internodes length/stem, single stalk weight, individual clump

weight, millable cane/clump, and stalk density. Biochemical properties of juice for

SEDPs and SETDPs were almost similar in SETDPs and SEDPs. Significant difference

between SEDPs and SETDPs was noted only in HR brix% of juice. Somaclonal

variations occurred among the plants regenerated through somatic embryogenesis

(SEDPs) could be used in breeding programme for the improvement of sugarcane

cultivars. 

Key words:  Embryogenesis, Somaclonal variation, Sugarcane.

 

INTRODUCTION

Sugarcane (Saccharum officinarum L.) is one of the economically important crops

widely cultivated in the tropics to subtropics and annually provides around 60 to 70 % of

the world’s sugar (Shah et al., 2009). It is an important food-cum cash crops and the

only source of sugar production in Bangladesh. It has long been observed that the

sugarcane varieties tend to run-out or decline in yield after a few years in a particular

area (Khan et al., 2009).The use of tissue culture for creation of somaclonal variation

can be used to increase the speed or efficiency of the breeding process to improve the

accessibility of existing germplasm of sugarcane and create new variation for crop

improvement. It has been recognized that all plants regenerated from tissue culture are

not exact replicas of a parental form and exhibit great variability in agronomic traits

(Heinz et al., 1977; Larkin and Scowcroft, 1983; Ramos Leal and Maribona, 1991). This

genetic alternation termed somaclonal variation (Larkin and Scowcroft, 1981), which is

being exploited to shorten the time needed to produce new breeding lines with

desirable traits. The somaclonal variation may be attributed to either (i) pre-existing

variation in the somatic cells of the explant (genetic) or (ii) variation generation during

tissue culture (epigenetic). Often both factors may contribute. Factors such as explants

source, explant age, duration of culture, number of sub-cultures, culture environment,

chemical additives or growth stimulants or regulators, media composition, the level of

ploidy and genetic mosaicism are capable of inducing in vitro variability (Silvarolla,

1992).  Plant regeneration from calli, cells, protoplast have been shown to exhibit great

variability in agronomic traits reported in potato (Shepard et al., 1980), rice (Oono,

1978), maize (Green, 1977) and even in sugarcane (Heinz et al., 1977; Larkin and

Scowcroft, 1993 and Ramos Leal and Maribona, 1991). In sugarcane somaclones have

been identified with increased resistance to both Fiji and downy mildew disease

(Krishnamurthi, 1974; Krishnamurthi and Tlaskal, 1974) and eyespot disease (Ramos

Leal et al., 1996). Salt tolerance somaclones have also been generated by a tissue

culture cycle (Samad and Begum, 2000 and Khan et al., 2004). The present study was

undertaken with a view to evaluate the somaclonal variation in somatic embryo derived

plants of sugarcane var. Isd-16 which can be used in local agronomic field for mass

production with efficient output.

Methods and Materials

Plant material

Young shoot apics of 3, 6 and 12 months field grown clones of sugarcane var Isd-16

were used as the source of explants. The explants were provided by the Bangladesh

Sugarcane Research Institute (BSRI), Ishurdi, Pubna and maintained at the

experimental field of Botanical Garden, Department of Botany, University of Rajshahi,

Bangladesh.

Culture conditions

The pH of all media adjusted to 5.7 and autoclaved at 15 psi for 20 min at 1210C. Initial

cultures i.e. serial slices were cultured on agar (8mg/L) solidified medium in test tubes.

After shoot induction the explants were transferred onto MS medium in jars containing

agent. All cultures were incubated in a culture room at 25±20C under cool white

fluorescent light (60 µE/m2/s) with a 16 h photoperiod. Sub-culturing was carried out

every fortnightly.

Callus induction and multiplication

Apical portion of healthy shoots were stripped to the terminal bud and attached

immature leaf rolls were surface sterilized with 0.1% HgCl2 for 9 min. Approximately 3

slices (5 x 3 mm) were taken from each cylindrical leaf roll and cultured on MS medium

(Murashige and Skoog, 1962) supplemented with 1-5 mg/L 2,4-D alone and incubated

in dark for callus induction. When the callus was abundant, it was transferred to the

regeneration medium.

Protocol for somatic embryogenesis and shoot proliferation

Plantlets were regenerated through somatic embryogenesis from the embryogenic

callus derived from leaf sheath explants of sugarcane var. Isd-16 using the protocol

developed by Roy (2006). The embryogenic calli with induced shoot were transferred to

MS liquid medium supplemented with various concentrations of BAP (0-2mg/L) for

shoot proliferation and multiplication.

Field evaluation of somaclones

The plantlets after proper acclimatization were transplanted in the experimental field of

Botanical Garden, Department of Botany, University of Rajshahi, during 2004.

Conventional setts derived plants (SETDPs) also planted side by side as control. The

experiment was laid out in randomized block design with three replications. Each

replication consisted 5 x 3 m plots. Row to row and plant to plant distance were 1 m

and 0.3 m respectively. To evaluate the incidence of somaclonal variation among

somatic embryo derived field grown plants (SEDPs) and their control (SETDPs), data

on different qualitative and quantitative parameters were recorded after 8-12 months of

planting.

Biometric parameters

The biometric parameters viz. shoot habit, tillering habit, tillering density, auricle, legule,

stalk colour, bud shape, stalk height, tillers/plant, internodes length/stem, single stalk

weight, individual clump weight, millable cane/clump, and stalk density of both the

somatic embryos derived plants (SEDPs) and setts derived plants (SETDPs) were

taken at the age of 12 months.

Biochemical parameters

The biochemical parameters viz. brix per cent juice, pol per cent juice, purity per cent

juice, pol per cent cane, recovery percent and RS (Reducing Sugar) percent were

estimated in the laboratory using CSR method. The analysis was performed on six stalk

samples which were collected at the age of 12 months. The stalks were shredded using

a cutter grinder and juice squeezed with a hydraulic press at 60 lbs/sq.inch.

Statistical analysis

Student ‘t’ test was performed to find out the difference between the individual sett

derived plants (SETDPs) compared to somatic embryo derived plants (SEDPs) for

biometric and biochemical parameters.

Results

Callus induction

Explants showed variation for callus development in MS medium with different

concentrations of 2,4-D. 3 month old immature leaf sheath explants produced highest

calli (51%) in MS medium supplemented with 4 mg/L 2,4-D. Explants of 6 and 12 month

old leaf sheath developed optimum calli (91 and 62%, respectively) in MS medium

fortified with 3mg/L 2,4-D.

Differentiation and somatic embryogenesis

In the present study, Optimum (42%) somatic embryo induction from immature calli was

obtained in MS medium supplemented with 0.5 mg/L 2,4-D. Callus underwent

embryogenesis producing huge number of somatic embryos when sub-cultured on MS

medium supplemented with 25 mg/L L-proline. Plant regeneration was achieved (98%)

by transferring them onto half-strength MS0. The highest shoot multiplication was

achieved when BAP was at a concentration of 1.5mg/L in MS medium. Plantlets with

well-developed roots were transferred into poly bags contained compost for primary

acclimatization and then transplanted to the field.

Field evaluation of somaclones

In the current investigation, the researchers found that a large number of somatic

embryo derived plants (SEDPs) were morphologically different with some distinct

characters as compared to setts derived plants (SETDPs) (Table1). Variations in almost

all qualitative characters were noticed between SEDPs and SETDPs. The highest 60%

of SEDPs was exhibited variation in stalk colour. Among them 33% had brownish green

stalk, 20% had yellowish green and 7% had reddish green. Profuse tillering density was

another exceptional character found in SEDPs and 53% plants possessed this

character. Similar percentage (33%) of SEDPs showed variations in terms of outer

auricle and bud shape. Plant habit, tillering and internodes habits of some SEDPs were

also different from SETDPs. Results also reveals that occurrence of different types of

somaclonal variants were more frequent among SEDPs populations. Occurrence of

dwarf off-types plant was very common. Other morphological characteristics such as

short leaves, deformed internodes and semi-erect stem were found to be linked with

dwarfism.

Present findings showed that SEDPs of sugarcane var. Isd-16 possessed significant

differences from setts derived plants (SETDPs) populations as control in the

characteristics of stalk height, tillers/plant, five internodes length/stem, individual clump

weight, millable cane/clump, and stalk density.

Biochemical properties of juice for SEDPs and SETDPs were almost similar in SETDPs

and SEDPs (Table 2 and 3). Significant difference between SEDPs and SETDPs was

noted only in HR brix% of juice. These results of occurrence of somaclonal variations in

SEDPs populations in sugarcane var. Isd-16 due to tissue culture are more or less

concomitant with the previous studies. Moreover, these results elucidated those genetic

variations of sugarcane var. Isd-16 could open an opportunity for selection of elite

clones.

Discussion

Callus induction

Immature sugarcane leaves are established good explant source for callus production

(Brisbe et al., 1994; Chengalrayan & Gallo-Meagher, 2001, Shah et al., 2009) and has

been used to induce callus from various sugarcane explants (Oropeza and Garcia,

1996; Gallo-Meagher et al., 2000, Shah et al., 2009). Callus induction in Ms medium

fortified with 2,4-D was reported by Chen et al (1988). Behera and Sahoo (2009)

reported the highest percentage of callus induction in MS medium supplemented with

2.5 mg/L 2,4-D. High amount of calli (100%) were produced using 2,4-D in the

concentration of 3 mg/L with MS medium by Ather et al (2009).

Differentiation and somatic embryogenesis

Induction of somatic embryogenesis was reported by many workers in MS medium

supplemented with 2,4-D and coconut water (Ahloowalia and Maretzki, 1983; Ho and

Vasil, 1983) and  in MS medium  fortified with proline, ABA and 2,4-D (Gosal et al,

2003). Higher shoot regeneration was achieved on a medium containing only BAP at

concentration of 0.5 mg/L (Gosal et al, 2003).

Field evaluation of somaclones

Heinz and Mee (1971) found variations among callus-derived H 50-7209-14-24 plants

when compare to their control with respect to tops, dewlap, auricle shape, leaf sheath

colour and hair group.  Sood et al. (2006) demonstrated that tissue culture derived

sugarcane var. CoJ 64 plants attained better height, millable cane height, a greater

number of live buds, increased cane yield and sugar recovery % as compared to

conventionally propagated sugarcane plants under parallel agronomic practices in the

field. They also reported that high tillering is resulted in thinner canes because

thickness of the canes is directly proportional to the number of tillers per clump and is

also related to the cytokinin effect. The tissue-cultured plants in the first year were

thinner but the thickness was increased in first ratoon and subsequent generations. Lat

and Lantin (1976) reported that some of the somaclones of sugarcane cultivars CAC

57-13 showed significant differences from the parental variety in cane diameter stalk

length and weight. Liu and Chen (1976, 1978 a, b) found significant variation amongst

sugarcane somaclones from eight varieties in characters such as cane yield, sugar

yield, stalk number, length, diameter, volume, density and weight, per cent fiber, auricle

length, dewlap shape, hair group and attitude of top leaf. Some of this somaclones

showed significant improvements over the parental performance. Siddiqui et al. (1994)

compared the brix % of canes of somaclones with those of their parents and found the

somaclones were better than their parents in this character. On the other hand, Khan et

al. (2004) reported that brix % of canes of somaclones was less than those of their

parents. They also reported that the somaclones were found better in the characters of

tillers/plant, stalk height, number of nodes/stem and root band width but they found no

difference in the length of internodes of somaclones and source plants.

The original ploidy level of the plant or the plant organ from which the explants were

taken may play an important role in such somaclonal variations. Non-meristematic

explants generally produce genetic variability as compared to meristamatic explants.

However the cells in non-meristematic explants are the derivatives of meristematic part

of the plant and during their subsequent dedifferentiation, gross changes in their

genome may be arise including endopolyploidy, polyteny and amplification or

diminuition of DNA sequences (D’ Amato, 1989). Moreover, when the cells were

various genomic constitution of the initial explants are induced to divide in culture; the

cells may exhibit changes in chromosome number such as aneuploids and polyploids

but very often from these mixoploid callus cultures.

The culture environment specially the choice and particularly by the concentration of

growth regulators in the medium is influenced the somaclonal variation (Karp, 1992). It

is possible that growth regulators act as mutagens. The synthetic auxin (2,4-D) has

been shown to increase the frequency of blue to pink mutation in the Tradescantia

stamen hair system (Dolezel and Novak, 1984) and to induce significant increases in

the frequency of sister chromatid exchanges in root tip cells of Allium sativum (Dolezel

et al., 1987). However, there is a paucity of examples of this kind and most evidence

points to growth regulators influencing somaclonal variations during the culture phase

through their effects on cell division (Gould, 1984), the degree of disorganized growth

(Karp, 1992), and selective proliferation of specific cell types (Ghosh and Gadgil, 1979).

Conclusion

The cause of the morphological, agronomical and biochemical variations observed in

the present study, though not well defined but it may be linked with the use of synthetic

auxin (2,4-D) and inherent chromosomal instability in callus culture. Larkin et al. (1985)

evidenced that transposable elements play a vital role on somaclonal variation.

Whereas, Siddiqui et al. (1994) mentioned that one did not know the phase at which the

variation arise. According to them the variations are caused by a combination of

physical and chemical phenomenon. In the present study, somaclonal variations

occurred among the plants regenerated through somatic embryogenesis (SEDPs) also

could be used in breeding programme for the improvement of sugarcane cultivars.

Acknowledgements

We would like to extend thanks to the BSRI, Ishurdi, Pubna for providing researc

materials, laboratory facilities for conducting biochemical analysis. Special thanks to Dr.

Md. Amzad Hossain, Associate Cane Nutritionist, BSRI, Ishurdi, Pubna for giving

necessary information during the research work.

References

Ahloowalia BS and Maretzki (1983). Plant regeneration via somatic embryogenesis in

sugarcane. Plant cell rep., 2(1): 21-25.

Ather A, Khan S, Rehman A and Nazir M (2009). Optimization of the protocols for the

callus induction, regeneration and acclimatization of sugarcane cv. Thatta-10.Pak J

Bot., 41(2): 815-820.

Bahera K K and Sahoo S (2009). Rapid in vitro micropropagation of sugarcane

(Saccharum officinarum L. cv-Nayana) through callus culture. Nature and science, 7

(4):1-10.

Chen WH, Davey MR, Power JB and Cocking EC(1988). Control and maintenance of

plant regeneration in sugarcane callus cultures. J Exp. Bot., 39: 251-261.

D’Amato F (1989). Polyploidy in cell differentiation. Caryologia. 42: 183-211.

Dolezal J and Novak FJ (1984). Effect of plant tissue culture media on the frequency of

somatic mutations in Tradescantia stamen hairs. Zpflanzenphysiol. 114: 51-58.

Dolezal J, Lucretti S and Novak FJ (1987). The influence of 2,4- dichlorophenoxyacetic

acid on cell cycle kinetics and sister chromatid exchange frequency in garlic (Allium

sativum) meristems cells. Biologia Plantarum. 29: 253-257.

Ghosh A and Gadgil VN (1979). Shift in ploidy level of callus tissue: A function of

growth substances. Indian J. Exp. Biol. 17: 562-564.

Gill N K, Gill R and Gosal SS (2003). Factors enhancing somatic embryogenesis and

plant regenartion in sugarcane (Saccharum officinarum L.). Indian J Biotech., 3: 119-

123.

Gould AR (1984). Control of the cell cycle in cultured plant cells. C.R.C. Critical Rev.

Plant Sci. 1: 315-344.

Green GE (1977) Prospects for crop improvement in the field of cell culture. Hort. Sci.

12: 7-10.

Heinz D M, Krisshnamurti L, Nickel and Maretzki A (1977). Cell, tissue and organ

culture in sugarcane improvement. In Reinert J and Bajaj Y P S (eds). Applied and

Fundamental Aspect of Plant Tissue and Organ Culture. Springer- Verlag, New York.

pp. 13-17.

Heinz DJ and Mee GWP (1971). Morphologic, cytogenetic, and enzymatic variation in

Saccharum species hybrid clones derived from tissue culture. American J. Bot. 58 (3):

257-262.

Hendre R R, Iver R S and Kotwal M (1983). Rapid multiplication of sugarcane by tissue

culture. Sugarcane 1: 5-8.

Ho W-J and Vasil I K (1983). Somaic embryogenesis in sugarcane (Saccharum

officinarum L.): Growth and plant regeneration from embryogenic cell suspension

cultures.annals of BOt., 51:719-726.

Karp A (1992). The role of growth regulators in somaclonal variation. In Miflin B J (ed.)

Oxford Surveys of Plant Molecular and Cell Biol. Vol. 7. Oxford University Press. pp. 1-

58.

Khan S J, Khan M A, Ahmad H K, Khan R D and Zafar Y (2004). Somaclonal variation

in sugarcane through tissue culture and subsequent screening for salt tolerance. Asian

J. Plant Sci. 3 (3): 330-334.

Khan IA, Dahot M U, Seema N, Yasmin S, Bibi S, Raza S and Khatari A, (2009).

Genetic variability in sugarcane plantlets developed through in vitro mutagenesis. Pak J

Bot.,42(1): 153-166.

Krishnamurthi M and Tlaskal J (1974). Fijii disease resistant Saccharum officinarum

var. pindar subclones from tissue cultures. Proc. Int. Sco. Sugar Cane. Technol. 15:

130-137.

Larkin P J and Scowcroft W R (1983). Somaclonal variation and eye spot toxin

tolerance in sugarcane. Plant Cell Tiss. Org. Cult. 2: 111-121.

Larkin P J, Brettell R I S, Ryan S A, Davies P A, Pallotta M A and Scowcroft W R

(1985). Somaclonal variation: Impact on Plant Biology and Breeding Strategies. In.

Zaitlin M, Day P and Hollander A (Ed.). Biotechnology in Plant Science: Relevance to

Agriculture in the Eighties. Academic Press, New York. pp. 83-100.

Larkin PJ and Scowcroft WR (1981). Eye spot disease of sugarcane. Induction of host

specific toxin and its interaction with leaf cells. Plant Physiol. 67: 408-414.

Larkin PJ and Scowcroft WR (1981). Somaclonal variation-a novel source of variability

from cell cultures for plant improvement. Theor. Appl. Genet. 60: 197-214.

Larkin PJ and Scowcroft WR (1983). Somaclonal variation and eyespot toxin tolerance

in sugarcane. Plant Cell Tiss. Org. Cult. 2: 111-121.

Lat J B and Lantin M M (1976) Agronomic performance of sugarcane clones derived

from callus tissue. Philippine J. Crop Sci. 1: 117-123.

Liu M C and Chen W H (1976). Tissue and cell culture as aid to sugarcane breeding .1.

Creation of genetic variation through callus culture. Euphytica 25: 393-402.

Liu M C and Chen W H (1978a). Tissue and cell culture as aids to sugarcane breeding.

2. Performance and yield potential of callus derived lines. Euphytica 27: 273-282.

Liu M C and Chen W H (1978b). Improvement in sugarcane by using tissue culture

methods. (Abst) Fourth Intl. Congr. Plant Tiss. Cell Cult., Calgary, Canada. pp. 163.

Murashige T and Skoog FA (1962). A revised medium for rapid growth and bioassays  

with tobacco tissue cultures. Physiol. Plant. 15: 473-497.

Oono K (1978). Test tube breeding of rice by tissue culture. Trop. Agric. Res. Series.

11: 109-123.

Ramos Leal M A, Ruiz I, Sandoval and Maribona R H (1991). Effect of DS toxin upon

the permeability of sugarcane leaf and callus tissue. Plant Breeding. 107: 242-247.

Ramos Leal MA, Maribona RH, Ruiz A, Korneva S, Canales E, Dinkova TD, Izquierdo

F, Coto O and Rizo D (1996). Somaclonal variation as a source of resistance to

eyespot disease of sugarcane. Plant Breeding. 115: 37-42.

Roy M (2006). Somatic embryogenesis of sugarcane (Saccharum officinarum L.) and

its transformation by Agrobacterium. Ph D Thesis, University of Rajshahi, Rajshahi.

Samad MA and Begum S (2000). Somaclonal variation of Nonirradiated and Irradiated

calli of sugarcane ((Saccharum officinarum L.). Plant Tiss. Cult. 10 (1): 25-29.

Shah A H, Rashid N, Haider M S, Saleem F, Tahir M and Iqbal J. (2009). An efficient,

short and cost-effective regeneration systemfor transformation studies of sugarcane

(Saccharum officinarum L.). Pak J. Bot.. 42(2): 609-614.

Sharp WR, Sohndahl MR, Evans AE, Caldas LA and Maraffa SB (1980). The

physiology of in vitro asexual embryogenesis. Horticultural reviews. 2: 268-310.

Siddiqui S H, Khatri A, Javed M A, Khan N A and Nazamani G S (1994). In vitro culture.

A source of genetic variability and an aid to sugarcane improvement. Pak. J. Agric.

Res. 15: 127-133.

Silvarolla M B (1992). Plant genomic alternations due to tissue culture. J. Brazil. Assoc.

Adv. Sci. 44: 329-335.

Sood N, Gupta P K, Srivastava R K and Gosal S S (2006). Comparative studies on field

performance of micropropagated and conventionally sugarcane plants. Plant Tiss. Cult.

& Biotech. 16 (1): 25-29.

Sreenivasan T S and Sreenivasan J (1992). Micropropagation of sugarcane varieties

for increasing cane yield. SISSTA Sugar. J. 18: 61-64.

Table 1. Morphological characters of conventional setts derived (SETDPs) and somatic

embryo derived plants (SEDPs) of sugarcane var. Isd-16. Data were recorded 7-8

months after planting.

 

- = No variation

Table 2.  Mean performance for stalk characters of sett derived (SETDPs) and somatic

embryo derived plants  (SEDPs) of sugarcane var. Isd-16 after 12 months of planting.

 

 

Characters

Sett derived plants

(SETDPs)

Somatic embryo

derived plants

(SEDPs)

Somaclonal variants

Exceptional char. in

SEDPs

 

%

         

Plant habit Erect Erect and semi-erect Semi -erect 13

Tillering habit Compact Compact and

spreading

Spreading 20

Tillering

density

Intermediate Intermediate and

profuse

Profuse 53

Leaf blade Green, curved near

tip

 

do

 

-

 

-

Outer auricle Straight transitional Straight transitional

and deltoid

Deltoid 33

Legule Crescent with broad

lozenge

Crescent with broad

lozenge and Broad

crescent

Broad crescent 13

Stalk colour  

Green, greenish

yellow

Green, greenish

yellow, yellowish

green, brownish green,

reddish green

Brownish green,

yellowish green,

reddish green

 

60

Internode habit Cylindrical Cylindrical, bobbin and

conoidial

Bobbin and

conoidial

20

Bud Simple ovate Simple or narrow ovate Narrow ovate 33

 

 

 

Table 3. Mean performance for Juice characters of sett derived (SETDPs) and somatic

embryo derived plants (SEDPs) of sugarcane var. Isd-16. Data were recorded from 12-

months-old plants.

 

 

Characters

Setts derived plants

(SETDPs)

Somatic embryo derived

plants (SEDPs)

 

t value Range Mean ± SE Range Mean ± SE

           

Stalk

height

(cm)

198-250 221.05 ± 4.94 160-280 247.25 ± 6.50 **

Stalk girth

(cm)

2.0-2.5 2.15 ± 0.04 1.7-3.0 2.08 ± 0.74 ns

Nos. of

nodes/stem

20-28 24.75 ± 0.37 18-28 23.05 ± 0.63 ns

Five

internodes

length

(cm)/stem

40-60 51.49 ± 1.20 46.5-70 55.25 ± 1.37 *

Tillers/plant

(nos.)

5-8 6.25 ± 0.20 4-14 8.5 ± 0.55 **

Single

stalk

weight

(kg)

0.52-0.80 0.63 ± 0.02 0.53-0.80 0.61 ± 0.02 ns

Individual

clump

weight

(kg)

1.2-3.12 2.45 ± 0.12 1.74-4.0 2.80 ± 0.10 *

Millable

cane/clump

(nos.)

2-5 3.90 ± 0.19 3-5 4.55 ± 0.13 *

Stalk

density

(c.c)

0.55-1.12 0.73 ± 0.38 0.52-1.15 0.87 ± 0.40 *

 

Characters

Sett derived plants (SETDPs) Somatic embryo derived plants

(SEDPs)

 

t value Range Mean ± SE Range Mean ± SE

ns = non significant

* = significant at 5% level

** = significant at 1% level

 

ns = non significant

* = significant at 5% level

Figure legends

Figure 1. In vitro regenerated sugercane plant var. Isd-16 developed through somatic

embryogenesis.

Figure 2. Field grown micropropagated sugarcane var. Isd-16 after 2 months of

planting.

Figure3. Field grown micropropagated sugarcane var. Isd-16 after 12 months of

planting.

Figure 4. Internode, girth and colour differences among somatic embryo derived plants.

 

 

 

 

 

 

 

 

 

 

 

HR brix % juice 14.48-23.0 19.20 ± 0.49 19.0-23.0 20.55 ± 0.29 *

Pol % juice 11.88-17.87 14.97 ± 0.40 12.07-17.88 14.75 ± 0.42 ns

Purity % 57.41-89.30 74.92 ± 0.48 58.99-89.38 71.05 ± 2.58 ns

Recovery % 4.74-10.91 7.96 ± 0.47 4.74-10.91 7.25 ± 0.49 ns

Pole % cane 7.24-13.94 10.33 ± 0.56 7.24-13.94 9.91 ± 0.55 ns

R.S. % 0.86-3.0 1.83 ± 0.20 0.86-3.0 2.24 ± 0.12 ns

 

 

 

  

 

       

                                         Figures 1-4. Roy et. al.,2010