Gene expression analysis of MaMADS2, a ripening control mads-box gene, in Cardaba (BBB, Musa balbisiana Colla)

Date

4-2014

Degree

Bachelor of Science in Agricultural Biotechnology

College

College of Agriculture and Food Science (CAFS)

Adviser/Committee Chair

Eureka Teresa M. Ocampo

Abstract

Abstract: The study was conducted to quantify and analyze the gene expression of the ripening specific MADS-box gene, MaMADS2, in Cardaba during ripening particularly at the green mature, climacteric, 70% yellow and 100% yellow stages. Fruit RNA was extracted from banana pulp and reverse transcribed to cDNA. Regular reverse transcription PCR was used to verify the expression of MaMADS2 gene using specific primers that spanned the open reading frame. Based on visual estimation of the band intensities, regular PCR showed that the expression of MaMADS2 gene was highest during the climacteric stage and was least when fruits where 70% yellow. Real-time PCR analysis or qPCR was employed to verify the observed gene expression. Real-time PCR conditions were optimized, with starting cDNA material normalized to 50ng/μL. In addition two genes, actin and elongation factor alpha were screened for possible use as reference genes for qPCR analysis. Several qPCR runs were performed and results showed that absolute gene expression levels were lowest during the green mature stage, increasing dramatically during the climacteric, followed by a decrease in expression at 70% yellow stage, then another increase at the 100% yellow stage. The trend in MaMADS2 gene expression is similar to the biphasic increase in ethylene biosynthesis observed in bananas but not in other fruits. In addition, the expression of MaMADS2 even during the green mature unripe stage shows that MaMADS2 is expressed before ethylene biosynthesis, and points to its regulatory role on ethylene biosynthesis and onset of fruit ripening.

Language

English

Location

UPLB Main Library Special Collections Section (USCS)

Call Number

LG 993.5 2014 A127 S87

Document Type

Thesis

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