ISHS Acta Horticulturae 618: XXVI International Horticultural Congress: Environmental Stress and Horticulture Crops
TOWARDS THE IDENTIFICATION OF CANDIDATE GENES INVOLVED IN THE SIT TOMATO (LYCOPERSICON ESCULENTUM MILL.) MUTANT BY USING CDNA-AFLP ANALYSIS IN ISOGENIC LINES |
| Authors: | A. Sciancalopore, G. Martelli, I. Greco, F. Cellini, F. Sunseri, M. Gallitelli |
| Keywords: | Lycopersicon esculentum Mill., TDFs, transcript-derived fragments, ABA-deficient mutant, sitiens, GenEST |
| Abstract:
Amplified fragment length polymorphism (AFLP) analysis, a PCR based technique, is capable of detecting more than 50 independent loci in a single PCR reaction. The objective of the present study was to assess the polymorphic AFLP fragments between two isogenic lines (IL) of tomato (Lycopersicon esculentum Mill.), obtained from a backcross of an ABA-deficient mutant in a genetic background cv. “Money Maker”, both at genomic (gDNA) and cDNA level. Eighty primer pairs were tested on cDNA detecting a total of ~ 8.300 AFLP fragments in the IL, with an average of 100 fragments obtained per primer pair, 164 fragments (1.7% of the total observed) were polymorphic, between the genotypes tested. Thirty primer pairs tested on genomic DNA resulted in ~ 3.200 AFLP fragments (an average of 103 fragments per primer pair), only 9 fragments (0.28%) were polymorphic between the wild type and the isogenic ABA-deficient mutant sitiens. The AFLP primer pairs that show differential profiles between the ILs were used to isolate transcript-derived fragments (TDFs) in order to characterize cDNA clones potentially involved in the ABA pathway. Generation of ESTs as differentially expressed sequences, between IL, is proposed as a way of developing ESTs around specific trait for an integrated approach to isolate genes of interest. Starting from the 164 transcript-derived fragments (TDFs) detected, 23 fragments were isolated and sequenced. The obtained sequences, compared against public databases, showed similarity with several EST tomato sequences of unknown functions. | |
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