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Considering the environmental conditions of Pakistan where sugarcane breeding is constrained due to non viable fuzz (seeds) production. Somaclonal variation could prove to be a useful tool to overcome the difficulties in cane breeding. In the present
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   Pak. J. Bot ., 46(4): 1531-1535, 2014. SCREENING OF SUGARCANE SOMACLONES OF VARIETY BL4 FOR AGRONOMIC CHARACTERISTICS   SABOOHI RAZA* 1,2 , SYEDA QAMARUNNISA 1 , ISHRAT JAMIL 1 , BEENA NAQVI 3 , ABID AZHAR 1  AND JAVED A. QURESHI 1 1 The Karachi Institute of Biotechnology and Genetic Engineering (KIBGE), University of Karachi, Pakistan. 2  Nuclear Institute of Agriculture (NIA), TandoJam, Pakistan. 3 Pakistan Council of Scientific and Industrial Research (PCSIR), Karachi, Pakistan *Correspondence e-mail:; Tel # 092213013559827 Abstract Considering the environmental conditions of Pakistan where sugarcane breeding is constrained due to non viable fuzz (seeds) production. Somaclonal variation could prove to be a useful tool to overcome the difficulties in cane breeding. In the present study, 324 sugarcane somaclones regenerated from immature leaf roll callus of sugarcane variety BL4 were evaluated for their yield and yield contributing characters and the quality traits of cane. The field trial of somaclones showed variation in 160 somaclones from the mother plant in at least one character observed. Most of the somaclones showed variation in weight of stalks per plant; however, only twenty four out of 89 clones showed increase in the weight of the stalks per clump. The second highly variable trait was the number of stalks, 88 plants showed either increase or decrease in the number of stalks. It is noteworthy that the sucrose accumulation was not increased in any of the somaclones. Twenty one somaclones were selected for their increased yield potential. The comparative performance of these selected clones revealed that clones ‘K-250, K-265, K-251, K-109, K-106, K-300 and K-315 gave better sugar yield /plant as compared to BL4. Maximum sugar yield/plant was observed in Clone ‘K-250’ (2.5 Kg) followed by K-265 (2.44 Kg), whereas the average sugar yield of BL4 was 1.2 Kg/plant.   Introduction Sugarcane crop improvement in different countries relies on conventional breeding, mutation breeding, somaclonal variation and genetic engineering (Dalvi et al., 2012 ; Rajeswari et al ., 2009). Sugarcane improvement through conventional methods is time-consuming (Cox et al ., 2000), and is strictly dependent on the nature of flowering, viability of pollen, seed (Moore & Nuss, 1987; Khan et al.,  2008) and the genomic complexity of sugarcane crop (Ingelbrecht et al ., 1999). Considering the environmental conditions of Pakistan where sugarcane breeding is limited due to non viable fuzz (seeds), somaclonal variation presents an alternative solution to overcome many difficulties in cane breeding (Shahid et al ., 2011). An array of variations has been observed using tissue culture techniques in different crops (Nawaz et al ., 2013). This variation is termed ‘somaclonal variation’. Although somaclonal variation is undesirable for clonal propagation and genetic transformation efforts (Gao et al.,  2011, Pandey et al., 2012), it may serve as a useful tool in some crop improvement programmes (Evans et al ., 1984; Brown & Thorpe, 1995; Tiwari et al ., 2010). Thus far, for sugarcane, only a few improved variants have been released as cultivars after extensive efforts in different laboratories (Larkin & Scowcroft, 1983; Krishnamurthi & Tlaskal, 1974). As most of the agronomic important traits are quantitatively inherited in sugarcane, the frequency of  positive mutation in terms of high yield and increased sucrose content is very limited. Moreover, such variations are often unstable, which limits the ability of this  phenomenon to be used as a tool for crop improvement in sugarcane. (Kresovich et al. , 1986; Irvine et al. , 1991; Hoy et al.,  2003; Matsuoka & Giglioti, 2005). However, the frequency of phenotypic variation and the type of variation mostly depends on the genotype, explants used and the culture conditions. Pre-existing variability among the cells may play a major role in the frequency of somaclonal variation (Brown & Thorpe, 1995; Hoy et al.,  2003). Present study was conducted to evaluate the field  performance of regenerated somaclones for yield and yield contributing characters and the quality traits of cane in varity BL4. Materials and Methods R1 generation of 324 somaclones regenerated from immature leaf roll callus of sugarcane variety BL4 were evaluated for their yield and yield contributing characters and the quality traits of cane in the experimental field of Dr. A. Q. Khan Institute of Biotechnology and Genetic Engineering (KIBGE) University of Karachi using randomized complete block design (RCBD) with three replications. Plant height, cane diameter, number of internodes, length of internodes, leaf length, number of stalks/ stool and weight of stool were measured. Quality of the cane was estimated by measuring brix% (using hand refractometer, Alla France) and pol% (using  polarimeter, ATAGO, Japan) from extracted juice of cane. Fiber % was calculated as described by Thangavelu & Rao (1982). Commercial cane sugar percentage (CCS %) was calculated using Australian Commercial Cane Sugar (CCS) formula given by Meade & Chen (1977). Cane sugar recovery percent (CSR %) was calculated by the following formula: CSR % = Commercial Cane Sugar (CCS) % × 0.94 where, CCS is commercial cane sugar, and 0.94 is net titre (sugar losses) (Ghaffar et al. , 2011).  SABOOHI RAZA  ET AL ., 1532 One way analysis of variance (one way Anova) was done using SPSS version 17.0 and pair wise comparison of means of phenotypic traits of all somaclones with means of mother plant BL4 was done by calculating fisher’s least significant difference (LSD) at p< 0.05. Pearson’s correlation between all phenotypic traits and Duncan's multiple range test (DMRT) of the selected clones were done by SPSS version 17.0. Result and Discussion Out of all well grown somaclones in three replicates, 160 somaclones exhibited variations over its mother plant in at least one character observed in this study. Maximum variation was observed in weight of stalks per plant, 89 somaclones showed variation in this trait. Twenty four out of 89 clones showed increase in the weight of stalks per clump (Fig. 1). Reduced weight of stalk/plant was observed in 65 somaclones ranging from 21-97% decrease (Fig. 1). The second highly variable trait was the number of stalks, 88 plants showed either increase or decrease in number of stalks (Fig. 1). In 32 somaclones, substantial increase in the number of stalks was observed, and 56 clones showed decreased number of stalks. After weight of stalks/ stool and number of stalks/ stool, maximum variations were observed in brix% where 44 somaclones showed increase in the length of internodes and 43 somaclones showed decrease in the brix% ( Fig. 1). Decrease in the brix% observed was upto 35%. Khan et al ., (2004) and Roy et al ., (2010) also observed similar decrease in the brix% which was contradictory to the observation by Siddique et al ., (1994), where increased brix% was observed in some of the clones. Changes in the cane diameter were observed in 31 somaclones, where a minor (15-20%) increase in the diameter was observed in only 5 clones (K-26, K80, K178, K152 and K156) ranging from 3.34-3.5cm; where cane diameter of BL4 was 2.9±0.089 cm. Hoy et al.,  (2003) also observed smaller cane diameter and increased number of stalks in the plants regenerated from callus culture of immature leaf rolls. Higher number of internodes, greater length of internodes and smaller diameter was also reported  by Sood et al.,  (2006). Twenty eight clones showed variation in cane height and twenty somaclones showed variation in the number of internodes respectively (Fig. 1). Fig. 1. Number of significant variations (p<0.05) observed in eight different characters. Cane yield and cane quality are major parameters for evaluating the commercial sugarcane hybrids in sugarcane improvement programmes. The yield of sugarcane is a quantitative character dependent upon various traits (Ahmed et al ., 2010). Correlation studies in sugarcane are helpful in selecting for improved clones (Kadian et al ., 2006). Correlation studies reveal that the cane yield /  plant was positively correlated with number of stalks per  plant, cane diameter, cane height and numbers of internodes at p<0.01 (Table 1). Numbers of stalks/stool seems to be an independent factor that greatly influenced the weight/stool (Table 1); while Sood et al ., (2006) found a strong negative relationship between the number of stalks with its diameter in the somaclones of sugarcane var. CoJ 64, Roy et al ., (2010) reported that the thinner canes were more prominent in the first generation (R  0 ) which turned to thicker canes in subsequent generations. In this study, cane diameter was positively correlated with all traits studied except for the number of stalks. However, the cane height, and number of internodes had a  positive contribution in the cane yield but had a negative impact over the sugar yield. Brix% was only positively correlated with cane diameter at p<0.01 (Table 1), which in turn correlated with all other yield contributing factors except for number of stalks. As no significant improvement in the brix% was observed, the criterion for selecting superior clone compared to the mother plant was to find the clones that had the high yield potential but not at the cost of brix. It was found that the two traits, the number of stalks/stool and the cane diameter were two of the major yield contributory factors that did not negatively correlate with the brix% (Table 1). That is the reason why all the clones showing substantially greater number of stalks were selected except for clones ‘K-17, K-27, K-88, K-121, K-198, K-226, K-241, K-253, K-255, K-262, K-269, K285, K-296, where most of them showed no significant improvement either in the yield or the brix% , while clone ‘K-27, K-241 and K-253 had significant decrease in the Brix% and clone ‘K-226 and K-285 had overall significant decrease in the yield i-e weight /stool. Significant increase in the cane diameter was observed only in five clones as mentioned above out of which clone ‘K-80, K-178 and K-152’ had no significant difference in the weight/stool and in the brix% (Data not shown). For this reason these clones were not selected, while clone’K-293 had more promising yield potential due to increased height and slight increase in the cane diameter with no significant loss in Brix% was included in the selected clones. The evaluation of comparative performance of the selected clones through Duncan's multiple range test (DMRT) is given in Table 2. The biochemical assessment for the Commercial Cane Sugar and sugar recovery % by analysing pol % and fiber % reveals that K-106, K- 250, K- 251, K- 265, K-287,K- 293, K-300 and K-315 is comparable with the CCS % of BL4 (Table 2). Howerver, the clones ‘K-250, K-265, K-251, K-109, K-106, K-300 and K315 gave better sugar yield /plant as compared to BL4. Maximum sugar yield/plant was observed in Clone ‘K-250’ (2.5 Kg) followed by K-265 (2.44 Kg). The average sugar yield of BL4 was 1.2 Kg.  SCREENING OF SUGARCANE SOMACLONES OF VARIETY FOR AGRONOMIC CHARACTERISTICS 1533    SABOOHI RAZA  ET AL ., 1534 Table 1. Pearson correlation coefficients between the different agronomical traits of somaclones grown in the experimental field of KIBGE Weight of stalks/stool Length of internodesCane Height Number of internodesBrix% Number of stalks/stool Cane diameter Pearson Correlation 1 Weight of stalks/stool Sig. Pearson Correlation 0.189 **  1 Length of internodes Sig. 0.000 Pearson Correlation 0.198 **  0.359 **  1 Cane height Sig. 0.000 0.000 Pearson Correlation 0.120 **  0.134 **  0.685 **  1  Number of internodes Sig. 0.005 .001 0.000 Pearson Correlation -0.019 -0.156 **  -0.104 *  -0.027 1 Brix% Sig. 0.660 0.000 0.014 0.520 Pearson Correlation 0.775 **  0.062 0.069 -0.026 -0.004 1  Number of stalks/stool Sig. 0.000 0.148 0.109 0.542 0.926 Pearson Correlation 0.163 **  0.117 **  0.258 **  0.247 **  0.263 **  0.071 1 Cane diameter Sig. 0.000 0.005 0.000 0.000 0.000 0.098 ** Correlation is significant at 0.01 level * Correlation is significant at 0.05 level As discussed earlier, a number of phenotypic variations were observed in in vitro  regenerated plants for important agronomic traits an observation shared by other scientists (Heinz & Mee, 1969; Liu et al ., 1983; Chen, 1986) and has been utilized in the sugarcane crop improvement programmes (Nickell & Maretzki, 1969; Heinz, 1973; Krishnamurthi & Tlaskal, 1974). It is noteworthy that sugar recovery is the major output of the sugarcane, which was not improved in the experiments conducted, but some clones showed sugar recovery comparable to BL4 with increased yield potential through which the sugar yield/ hectare could be improved. Some of the clones (clones ‘K-250, K-265, K-251, K-109, K-106, K-300 and K315) produced during this study are worth pursuing in successive generations as they may hold the potential for enhanced yield. References  Ahmed, A.O., A. Obeid and B. Dafallah. 2010.   The influence of characters association on behavior of sugarcane genotypes ( Saccharum Spp) for cane yield and juice quality. World J.  Agric. Sci ., 6: 207-211. Brown, D.C.W. and T.A. Thorpe. 1995. Crop improvement through tissue culture. World J Microbiol Biotechnol. , 11: 409-415. Chen, Z.Y. 1986. A clone of sugarcane with thick stalk and high sucrose content regenerated from tissue culture. Scientia  Agriculturae Sinica ., 3: 90. Cox, M., M. Hogarth and G. Smith. 2000. Cane Breeding and improvement. In: (Eds.): M. hogath, P. Allsopp. Manual of cane growing.  Bureau of Sugar Experimental Station  Indooroopilly . Australia, pp. 91-108. Dalvi, S.G., V.C. Vasckar, A. Yadav, P.N. Tawar, G.V. Dixit, T. Prsad and R.B. Deshmukh. 2012. Screening of promising sugarcane somaclones for agronomic traits and smut resistance using PCR amplification of intertranscrip region (ITS) of Sporisorium scitaminae.   Sugar Tech ., 14: 68-75. Evans, D.A., W.R. Sharp and H.P. Medina-Filho.1984. Somaclonal and gametoclonal variation.  Am. J. Bot.,  71: 759-774. Gao, D., B. He, Y. Zhou and L. Sun. 2011. Genetic and molecular analysis of a purple sheath somaclonal mutant in  japonica rice. Plant Cell. Rep ., 30: 901-911. Ghaffar, A., Ehsanullah,. Akbar, N. and Khan, S. H. 2011. Influence of zinc and iron on yield and quality of sugarcane  planted under various trench spacings. Pak J Agri Sci. 48: 25-33. Heinz, D.J. 1973. Sugarcane improvement through induced mutations using vegetative propagules and cell culture techniques. In:  Induced mutations in Vegetatively Propagated Plants . IAEA, Vienna. Austria. S11/PUB/, pp. 399: 53-59. Heinz, D.J. and G.W.P. Mee. 1969. Plant differentiation from callus tissue of Saccharum  species. Crop Sci ., 9: 346-348. Hoy, J.W., K.P. Bischoff, S.B. Milligan and K.A. Gravois. 2003. Effect of tissue culture explant source on sugarcane yield components.  Euphytica , 129: 237-240. Ingelbrecht, I.L., J.E. Irvine and T.E. Mirkov. 1999. Post transcriptional gene silencing in transgenic sugarcane. Dissection of homology-dependent virus resistance in a monocot that has a complex polyploid genome. Plant Physiol ., 119: 1187-1198. Irvine, J.E., G.T.A. Benda, B.L. Legendre and G.R. Machado. 1991. The frequency of marker changes in sugarcane plants regenerated from callus culture. Plant Cell. Tiss. Org. Cult  ., 26: 115-125. Kadian, S.P., R. Pal and Y.S. Lather. 2006. Correlation and path coefficient analysis in sugarcane.  Indian J. Agric. Res ., 40: 135-138.  SCREENING OF SUGARCANE SOMACLONES OF VARIETY FOR AGRONOMIC CHARACTERISTICS 1535 Khan, I.A., M.U. Dahot, N. Seema, S. Bibi and A. Khatri. 2008. Genetic variability in plantlets derived from callus culture in sugarcane. Pak. J. Bot  ., 40: 547-564. Khan, S.J., M.A. Khan, H.K. Ahmad, R.D. Khan and Y. Zafar. 2004. Somaclonal variation in sugarcane through tissue culture and subsequent screening for salt tolerance.  Asian J. Plant Sci ., 3: 330-334. Kresovich, S., R.E. McGee, H.J. Drawe and J.L. Rivera. 1986. Variability of agronomic characteristics in populations of tissue culture-derived and vegetatively propagated sugarcane. Proc Int Soc Sugar Cane Technol ., 19: 528-532. Krishnamurthi, M. and J. Tlaskal. 1974. Fiji disease resistant Saccharumoffinarum var. Pindar subclones from tissueculture. Proc Int Soc Sugarcane Technol . 15: 130-137. Larkin, P.J. and W.R. Scowcroft. 1983. Somacional variation and eyespot toxin tolerance in sugarcane. Plant Cell. Tiss. Org. Cult.,  2: 111-121. Liu, L.J., E.R. Marquez and M.C. Biascoechia. 1983. Variation in degree of rust resistance among plantlets derived from callus cultures of sugarcane in Puerto Rico. Phytopathology  Abstr  ., 73: 797. Matsuoka, S. and E.A. Giglioti. 2005. Breeding Strategies for sugarcane in 21st century: The challenges ahead. In: (Eds.): S. Solomon, S.S. Grewal, Y. Rui Li R.C. Magarey and G.P. Rao. Sugarcane: production, Management and Agroo-Industrial imperatives, pp. 485-506. IBDC. India. Meade, C. P. and Chen, J. C. P. 1977. Cane sugar hand book, 10thEdn. John Wiley and Sons, Inc. New York : 947. Moore, P.H. and K.J. Nuss. 1987. Flowering and flower synchronization. In: Sugarcane Improvement Through  Breeding , (Ed.): D.J. Heinz, pp. 273-311. Elsevier Amsterdam.  Nawaz, S., N. Ahmed, A. Iqbal and I. Khaliq. 2013. Optimization of regeneration protocols for wheat under drought and salt stress. Pak. J. Agri. Sci ., 50: 663-670.  Nickell, L.G. and A. Maretzki. 1969. Growth of suspension cultures of sugarcane cells in chemically defined media. Physiol Plantarum , 22: 117-125. Pandey, R.N., S.P. Singh, J. Rastogi, M.L. Sharma and R.K. Singh. 2012. Early assessment of genetic fidelity in sugarcane ( Saccharum officinarum ) plantlets regenerated through direct organogenesis with RAPD and SSR markers.  Australian Journal of Crop Sci., 6: 618-624 Rajeswari, S., S. Thirugnanakumar, A. Anandan and M. Krishnamurthi. 2009. Somaclonal variation in sugarcane through tissue culture and evaluation for quantitative and quality traits.  Euphytica ., 168: 71-80. Roy, M., M. Hossain, A. Biswas, R. Islam, S. R. Sarker and S. Akhter. 2010. Induction and evaluation of somaclonal variation in sugarcane ( Saccharum officinarum  L.) var. Isd-16. Gene Conserve , 9: 201-221. Shahid, M.T.H., F.A. Khan, A. Saeed and I. Fareed. 2011. Variability of red rotresistant somaclones of sugarcane genotype S97US297 assessed by RAPD and SSR. Genet  Mol Res ., 10: 1831-1849. Siddiqui, S.H., A. Khatri, M.A. Javed, I.A. Khan and G.S.  Nizamani. 1994.  In vitro  culture: A source of genetic variability and an aid to sugarcane improvement. Pak J.  Agric. Res ., 15: 127-13. Sood, N., P.K. Gupta, R.K. Srivastava and S.S. Gosal. 2006. Comparative studies on field performance of micropropagated and conventionally sugarcane plants. Plant Tiss. Cult Biotech ., 16: 25-29. Thangavelu, S. and K.C. Rao. 1982. Comparison of Rapi pol extractor and Cutex cane shredder methods for direct determination of fibre in Saccharum  clones. Proc. Ann. Conv. Sug. Tech. Assoc. India , 46: 15-21. Tiwari, A.K., Y.P. Bharti, S. Tripathi, N. Mishra, M. Lal, G.P. Rao, P.K. Sharma and M.L. Sharma. 2010. Review Article: Biotechnological approaches to improve sugarcane crop with special reference to disease resistance.  Acta Phytopathol Hun ., 45: 235-249. (Received for publication 5 March 2013)
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