Sandhu, Jagdeep SinghSharma, Amandeep Sahil2022-12-032022-12-032022Sharma, Amandeep Sahil (2022). Agrobacterium mediated transformation of sugarcane variety CO 0238 with chitinase gene for resistance against red rot (Unpublished M.Sc. thesis). Punjab Agricultural University, Ludhiana, Punjab, India.https://krishikosh.egranth.ac.in/handle/1/5810190287Endochitinase gene cloned from Trichoderma spp. was introduced into the sugarcane variety CO 0238 using Agrobacterium-mediated genetic transformation. The gene construct carrying endochitinase which was earlier mobilised into two Agrobacterium tumefaciens strains, LBA4404 and EHA105 independently, were used to agro-infect sugarcane spindle leaf roll segments. When spindle leaf rolls were agro-infected with LBA4404 strain a total of 45 plants were obtained from 326 agro-infected spindles and similarly 534 plantlets were obtained from 1911 agro-infected spindles with EHA105. The presence of transgene was confirmed by PCR, revealing amplicon size of 1275 bp corresponding to the endochitinase with transformation efficiency of 0.52% only with EHA105. RT-PCR revealed the differential expression of transgene in all PCR positive plants followed by quantitative relative expression analysis using qRT-PCR revealing up to 6-fold increase in endochitinase expression in seven sugarcane plants as compared to non-transgenic controls. This suggested that plant expressing higher expression can impart resistance against fungal pathogens, indicating the role of endochitinase in providing resistance against red rot.EnglishThesis