TRANSFORMATION OF POMEGRANATE (Punica granatum L.) BY AMP (ANTIMICROBIAL PEPTIDE) GENE TO CONFER RESISTANCE AGAINST BACTERIAL BLIGHT OF POMEGRANATE
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Date
2009-07-15
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University of Agricultural Sciences, Bangalore
Abstract
Pomegranate is a popular fruit crop and is of considerable
economic importance. The bacterial blight of pomegranate is becoming a
serious problem in major pomegranate growing area in India. All
conventional ways and means of controlling this disease have failed. In
pomegranate, existing protocols have shown slow response by
regeneration and less work has been reported on transformation. Hence,
in the present study, a systematic investigation was carried to
standardize an efficient in vitro regeneration protocol from different
explants of pomegranate cv. Bhagwa, find out the best treatment for
faster regeneration and transforming it with AMP gene. As a result, we
could able to achieve a reliable and efficient regeneration protocol and
standardize an efficient transformation protocol. Agrobacterium
tumefaciens carrying gene pCAMBIA construct with the constitutive
CaMV35S promoter, AMP gene terminator and nptII selectable marker
(Kanamycin resistance), was used for transformation of explants.
Putative transformants were identified on selection medium containing
kanamycin at different concentration. Transgene insertion and
expression at various levels were confirmed using PCR. Out of four
putative transformants analyzed, three transgenic plants showed PCR
+ve with AMP gene specific primer and further screening is going on
using different molecular techniques.
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