Cloning of vitellogenin gene of whitefly, Bemisia tabaci (Gennadius) into RNAi vector for genetic transformation of upland cotton
Loading...
![Thumbnail Image](assets/images/Item.jpg)
Date
2022
Authors
Journal Title
Journal ISSN
Volume Title
Publisher
Punjab Agricultural University, Ludhiana
Abstract
Cotton is India's important cash crop, and it plays a significant part in the country's industrial
and agricultural economy. Whitefly (Bemisia tabaci) has been one of the most threatening
polyphagous pests of cotton and no whitefly resistance germplasm is reported in India. Also,
there are no Bacillus thuringiensis (Bt) toxins known to be effective against whitefly.
Generating RNAi cotton plants expressing dsRNA of vital gene of whitefly can be a novel
approach to produce whitefly resistance in available Bt cotton variety. In present work,
208bp cDNA fragment of vitellogenin gene (VG) gene of whitefly was used to prepare an
RNAi construct. The cloning of RNAi cassette was confirmed through PCR with vector
specific primer sets. Agrobacterium harboring prepared RNAi construct was used to
transform cotton variety, PAU Bt3. The germinated cotton seeds containing
hypocotyledonary region of mature embryo was used as an explant (pricking and without
pricking) for Agro-inoculation for different incubation periods (90 and 120 minutes) and
thus, a cultivar-independent, in planta genetic transformation protocol has been
standardized. In transformation one cotyledon was removed followed by pricking with
needle on hypocotyledonary region resulted in 29 PCR (with RNAi cassette specific primers
in isolated DNA) positive RNAi cotton plants with overall cotton transformation efficiency
was 3.29%. PCR was also attempted in DNA isolated from all confirmed RNAi cotton plants
with Vir C gene specific primers which showed absence of Agro contamination in these
plants even after 5-6 months of transformation. These RNAi cotton plants will be further
characterized in future.
Description
Keywords
Citation
Ravinder Kaur (2022) Cloning of vitellogenin gene of whitefly, Bemisia tabaci (Gennadius) into RNAi vector for genetic transformation of upland cotton (Unpublished M.Sc. thesis). Punjab Agricultural University, Ludhiana, Punjab, India.