PJB-2019-272
Cloning, sequence characterization and expression analyses of ACO, ORFB and ORF25 genes from pepper cytoplasmic male sterility (CMS)
GUIFANG SHAO
Abstract
ORFB and ORF25, part of the F0 component of the F1F0-ATP synthase, always associated with energy metabolism. Aconitase is a vital enzyme for carbohydrate and energy metabolism, it is responsible for the interconversion of citrate and isocitrate in the tricarboxylic acid (TCA) cycle. Three genes complete coding sequence (CDS) were amplified using a reverse transcriptase PCR based on Online Software BLAST (https://blast.ncbi.nlm.nih.gov/Blast.cgi). The amplified fragment of ORFB is 750bp, but 471 bases is encoded in maintainer line of pepper, 462 bases in male sterile line; the amplified fragment of ORF25 is 835bp, but 588 bases is encoded in maintainer line and sterile line; the amplified fragments of the ACO is 2988bp in sterile line 9704A and maintainer line 9704B. Comparing to the species sequence downloaded from NCBI, the nucleotide sequence analysis showed that the amino acids were highly homologous with these species. The qRT-PCR showed that three genes have a different expression level between tissues in 9704B and flower in different developmental stages in two lines. These differences may cause pepper cytoplasmic male sterility.
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