Working group session:
Functional Genomics
Presentation type:
oral
Authors:
rashid, bushra; aftab, beenish; shahid, muhammad; hassan, Sameera; saddique, sajjad; husnain, tayyab
Presenter:
rashid, bushra
Correspondent:
rashid, bushra
Abstract:
CLCV is one of the most severe causes of cotton damage for which no adequate remedy is available. The possible solutions to this problem may be the transfer of resistant genes in the background of agronomically and technologically good varieties. In response to viral infection, several plants may show quantitative and qualitative differences in gene expression. In the present work differential display reverse transcriptase PCR (DDRT) was used to compare overall differences in gene expressions between infected and healthy plants. Screening through 12 primer-pair combinations resulted in up-regulation of 26 cDNA transcripts and 7 transcripts have significant homology with protein sequences of known genes i.e. ribosomal protein, Photosystem protein unit and a predicted hypothetical protein. The transcripts identified are first time reported against CLCV infection and submitted to GenBank database as novel ESTs.
The expression levels of 7 transcripts were evaluated by real-time RT-PCR. The GAPDH gene was used as the reference gene to normalize the expression levels. All transcripts showed different level of over-expression in healthy leaves as compared to CLCV diseased leaves. There is 1.6 fold more expression of predicted hypothetical protein, approx. 1 fold of ribosomal protein S14, 1 fold of photosystem reaction center subunit XI , 0.12 fold more expression of transcript A5B2, 0.62 fold more of uncharacterized membrane protein, 0.5 fold of transcript A5B1 as compared to the control.
Transcript A6B2 showed homology with Vitis vinefera protein that was supposed to be involved against CLCV infection, was selected and full length gene, GhLCVR (Accession Number: HQ338125) has been amplified. The gene comprises a single open reading frame of 142 amino acids with molecular mass of 16.1 kDa Predicted amino acid sequence shares 80-33% identities with Glycine max, Populus trichocarpa, Vitis vinifera, Zea mays, Ricinus communis, Arabidopsis thaliana, Picea sitchensis, Physcomitrella patens and Staphylococcus aureus respectively. This gene and other identified transcripts could provide a valuable source for the future improvement of cotton varieties against CLCV infection.