Submitted to: National Cotton Council Beltwide Cotton Conference
Publication Type: Abstract Only
Publication Acceptance Date: 1/4/2000
Publication Date: N/A
Technical Abstract: We used a novel approach to identify and isolate transcripts specific to the naturally occurring Ligon Lintless -1 mutant (Li-1). We used a PCR-based method to construct cDNAs from TM-1, near isogenic Li-1 and mutant F1 plants and bulked samples of the mutant F2 plants. A novel PCR-based method, using fluorescent-labeled primers from the flanking sequences of simple short repeats (SSRs), was used to study differential gene expression between the cDNAs. We also assayed the SSR markers using genomic DNAs from a similar set of plants except we included 5 normal and 5 mutant individual F2 plants. All the SSR primer pairs amplified genomic cotton DNAs while 52% amplified cDNA templates. There is no report, as per our knowledge, on the use of SSR-specific primers in the identification of functional plant genes. However, SSR specific primers had specific advantages in screening cotton cDNAs because they were generally polymorphic, provided very few bands thus easy to analyze, and the procedure is efficient, reproducible and does not require restriction digestion. Thus, it is a very simple PCR-based technique. We identified two transcripts polymorphic between TM-1 and Li- 1, one specific to Li-1 and the other specific to TM-1. The monomorphic banding patterns in the genomic DNAs in contrast to cDNAs with a Li-1-specific primers could be due to putative transcriptional regulation.