Agrobacterium mediated genetic transformation of Citrus reticulata cv. Khasi mandarin

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dc.contributor.advisorSingh, Salvinder
dc.contributor.authorBhandari, Sangeeta
dc.date.accessioned2022-07-12T07:40:31Z
dc.date.available2022-07-12T07:40:31Z
dc.date.issued2020-02
dc.description.abstractCitrus is number one fruit of the world on accounts of its high nutritional value. India is the fourth largest producer of Citrus in the world. The north-eastern region of India is a rich treasure of various Citrus species. Khasi mandarin is the most economically important one and plays a vital role in the socio-economic development of the people in this region. Khasi mandarins are declining at a very high rate due to its vulnerability to different pathogen and insect/ pest. Conventional breeding for overcoming these problems are limited in Citrus and are directly associated with the reproductive biology of Citrus. Recent advances in genetic engineering have made it possible to incorporate desirable genes from elite genotype mainly through Agrobacterium-mediated genetic transformation. Citrus species showed varied response to in vitro regeneration and genetic transformation. Cultivar specific optimization of in vitro regeneration and transformation protocol is very important. In the present investigation, in vitro regeneration and Agrobacterium mediated genetic transformation protocol for Khasi Mandarin was optimized using different explants like epicotyl, hypocotyl, nodal and inter nodal segment obtained from six-week-old in vitro grown zygotic seedling. Explants were transformed wih Agrobacterium strain LBA4404, harbouring plasmid pBI121-AtSUC-GUS containing nptII as a selectable marker and gus as a reporter gene. Hypocotyl was found to be the best explants for khasi mandarin transformation and regeneration. MS medium supplemented with BAP (2mg/L), NAA (0.5 mg/L), 2, 4-D (1mg/L), MES (0.5g/L), sucrose (30g/L) and acetosyringone (100μM) was found to be best medium for co-cultivation. Modified MS medium containing BAP (4mg/L), MES (0.5g/L), sucrose (30g/L), phytagel (4g/L), kanamycin (50mg/L) and timentin (150mg/L) showed highest regeneration efficiency (18%). Modified MS medium containing BAP (4mg/L), GA3 (0.5mg/L), MES (0.5g/L), sucrose (30g/L), phytagel (4g/L), kanamycin (50mg/L) and timentin (150mg/L) showed highest multiple shoot induction (6%). In vitro regenerated shoots that survived up to 3rd selection cycle were subjected to GUS assay for confirmation of GUS expression in the phloem tissues. Present investigation is a preliminary study for optimization of an in vitro regeneration and genetic transformation protocol in Khasi Mandarin.en_US
dc.identifier.urihttps://krishikosh.egranth.ac.in/handle/1/5810185398
dc.language.isoEnglishen_US
dc.pages53en_US
dc.subAgricultural Biotechnologyen_US
dc.themeAgrobacterium mediated genetic transformation of Citrus reticulata cv. Khasi mandarinen_US
dc.these.typePh.Den_US
dc.titleAgrobacterium mediated genetic transformation of Citrus reticulata cv. Khasi mandarinen_US
dc.typeThesisen_US
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