Cloning and Analysis of cDNAs Encoding Plastid-Lipid Associated Proteins in Bitter Gourd (Momordica charantia L.)
Date Issued
2008
Date
2008
Author(s)
Jiang, Siao-Huei
Abstract
Bitter gourd (Momordica charantia L.) is a monoecious species in Cucurbitaceae. Sex expression of bitter gourd is altered by exogenous application of plant growth regulators. Using 2- dimensional protein gel electrophoresis to identify sex expression-related proteins in bitter gourd, the results indicated that the plastid-lipid associated protein (PAP) expressed specifically during female flower development. To understand the mechanism of sex determination, plaque hybridization was performed to screen the PAP cDNA from bitter gourd library using Arabidopsis thaliana PAP gene as probe. Thirty-six cDNA clones were obtained. cDNA clone pMcPAP76 is 1,226 bp in length and possesses an open reading frame of 984 bp, enconding a polypeptide of 327 amino acids with an isoelectric point of 4.89 and a molecular weight of 35.53 kDa. The corresponding gene of pMcPAP76 cDNA is named McPAP1. The deduced amino acid sequence of McPAP1 shared high homology with other PAP around 62.6% to 79.9%, especially cucumber chromoplast protein C (CHRC) with 79.9% identity. Southern blot analysis indicated that McPAP1 belongs to a single-copy gene. Northern blot and RT-PCR analysis reflected that the expression of McPAP1 was specific in flower and fruit. The McPAP1 gene expressed most abundantly in male flowers at full blussom, but more abundantly at early stage in female flowers. The McPAP1 gene expression was consisted in the fruit development stage. The McPAP1 gene expression in fruit was found largest at 24-day after pollination which was 1.5 times as much as did that at 18-day after pollination. McPAP1 expression was induced in fruit tissues treated with ethylene, but inhibited by IAA. To clarify the McPAP1 localization and the relation to sex expression, GFP-fusion protein was constructed for transient analysis.
Subjects
Plastid-Lipid Associated Protein
Type
thesis
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