Induced systemic resistance and validation of post transcriptional gene silencing in tomato against tomato leaf curl virus
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UASD
Abstract
The efficacy of transgenic plants capable of PTGS was assessed in available T4 seeds
of four transgenic events viz., A, C, D and E carrying ihp-TRP subjected for PCR based
screening and gene segregation analysis. The bio-efficacy test of transgenic plants from three
events viz., ‘A’,’C’ and ‘E’ in T4 and T5 generation indicated moderate resistance against
ToLCV infection. The site of insertion of T-DNA carrying ihp-TRP genes was identified by
recovering the genomic sequences flanking right border of T-DNA obtained from TAILPCR,
identified the point of insertion on chromosome 1, chromosome 12 and chromosome 2
for ‘A’, ‘C’ and ‘E’ events respectively.
Thirty Pseudomonas and five actinobacteria isolates were functionally characterized
for ability of phosphate solubilization, nitrogen fixation, siderophore, indole-acetic acid and
gibberellic acid production. Out of thirty five, twenty isolates were prospected for control of
tomato leaf curl viral disease through seed priming, soil and foliar application to tomato
plants. Inoculation of five isolates, viz., AUDP326(4), AUDP360(2), AUDP139, AUDT217
and AUDT152 resulted in significant reduction in disease up to 60-80% and also could
significantly promote plant growth. Significant increase in the level of PAL, PO, chitinase
and phenolics were observed in inoculated plants compared to disease control. The SSH
studies at early (32-36 DAS) and late stage (40-46 DAS) in AUDT217 inoculated tomato
plants compared to uninoculated plants identified sixteen well annotated genes such as
protease inhibitor, threonine deaminase and other. However, SSH studies involving
inoculation of AUDT217 in the presence of ToLCV realized in the identification of twelve
well annotated transcripts of genes in the plant such as metallothionein-like protein,
ferredoxin-thioredoxin-reductase, metallocarboxypeptidase inhibitor and other which were
further validated through real time PCR studies.
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