Abstract
Callogenesis is a helping tool in tissue culture for creating variation. The study was carried out to investigate the response of different sugarcane varieties to callogenesis in order to analyze variation for sugarcane mosaic virus resistance. Three sugarcane accessions viz HSF-242, S2003-US-623, and S2003-US-633 were used in this study. The genotypes showed high value of callus score ranging from 2.59 to 3.25. Genotype HSF-242 was highest callus producer with an average score of 3.25. Three different media were used for organogenesis. S2003-US-633 proved to be the most responsive to organogenesis with an average of 43.33%. Material was exposed to shooting and rooting media and somaclones were shifted to green house. The somaclones produced were screened for sugarcane mosaic virus by ELISA (enzyme linked immunosorbent assay). Among 10 parent plants and 32 somaclones of HSF-242, only four somaclones showed susceptibility to SCMV disease while seven (7) parents showed positive reaction to the disease. Somaclones produced were also analyzed by microsattelite molecular markers to check the variation between parents and somaclones produced at molecular level. Molecular markers are useful for assessment of variation and phylogenetic relationships. Out of thirty (30) fragments amplified with fifteen (15) primers used among the sugarcane somaclones and parents, eleven (11) bands were polymorphic while rest of the nineteen (19) bands were monomorphic. Therefore, it may be concluded from the present results that SSRs can be used for identification of somaclonal variation and the relationship between parents and Somaclones.
Highlights
Callogenesis is a helping tool in tissue culture for creating variation
At the age of three months, both parents and somaclones were mechanically inoculated with sugarcane mosaic virus.Double Antibody Sandwich ELISA was performed according to method given by Kemeny and Challacambe (1989) for the screening of Sugarcane mosaic virus disease.In artificially virus, inoculated plants 10 parent plants and 32 somaclones of HSF-242 were selected for screening of virus through Das-ELISA test
The present study was undertaken to check the response of three sugarcane accessions viz., HSF-242, S2003-US-623, and S2003-US-633 for callogenesis, organogenesis, and presence of sugarcane mosaic disease and the existence of somaclonal variations by simple sequence repeats (SSR) marker
Summary
Callogenesis is a helping tool in tissue culture for creating variation. The study was carried out to investigate the response of different sugarcane varieties to callogenesis in order to analyze variation for sugarcane mosaic virus resistance. The recovery of sugar can be increased from the current average of 8.32% to 10/11 % by better cane varieties It is a photo-thermal sensitive crop and flowering takes place at 5-23° latitude whereas Pakistan is situated at 24-37° latitude. Callus culture of sugarcane have been successfully established using shoot apices, young leaves and young inflorescences as explant on MS medium containing 2, 4- D and coconut milk These clones show variation for different traits like high yield, more sugar recovery, disease resistance, drought tolerance and early maturity etc (Niaz & Quraishi, 2002a). This technique is used to make disease free plants in sugarcane by somaclonal variation. The existence of diseases in micropropagated plants of sugarcane can be checked by ELISA technique of virus detection, so it is very important to make disease free plants
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