Genetic transformation of guava (Psidium guajava L.) with RNAi construct for fruit fly [Bactrocera dorsalis (Hendel)] resistance
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Date
2020
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Punjab Agricultural University, Ludhiana
Abstract
Guava (Psidium guajava L.) is one of the most nutritious fruit due to its abundance in
antioxidants, tannins, vitamins A, C and minerals. But its production is severely affected by
fruit fly [Bactrocera dorsalis (Hendel)] infestation. There is no fruit fly resistant guava
germplasm reported so far globally. RNAi approach against fruit fly provides an attractive
approach to overcome this devastating problem. Here we report generation of marker-free
RNAi guava lines of cv. Allahabad Safeda targeting ecdysone receptor (ECR) gene of fruit fly
through in planta genetic transformation. For floral dip method, completely developed closed
bud and bud with calyx split were used and effects of different dipping time in infiltration
medium were observed. Dipping time for more than 10 seconds led to necrosis of floral buds
and pre-mature flower drop. For floral drop method, different amount of dropping inoculum
on bud with calyx split and open flower were used. Dropping 4-6 drops of Agrobacterium
inoculum led to necrosis and flower drop, while dropping 1-2 drops of inoculum found
optimum led to development of subsequent guava fruits. RNAi cassette integration was
detected in transformed guava lines through PCR using gene specific and vector specific
primers. No transformants were obtained with floral dip method in guava, but floral drop
transformation showed a higher transformation efficiency of 4.95% with bud with calyx split
stage of guava used as explant. The present optimized, efficient floral drop transformation
method will be very useful for genetic manipulation of guava.
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Gursimran Kaur (2020). Genetic transformation of guava (Psidium guajava L.) with RNAi construct for fruit fly [Bactrocera dorsalis (Hendel)] resistance (Unpublished M.Sc. thesis). Punjab Agricultural University, Ludhiana, Punjab, India.