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《Molecular Plant Breeding》 2018-10
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Cloning RCCR Gene from Mango(Mangifera indica) and the Construction of Gene Silencing Vector

Liu Kuanliang;Zhao Zhichang;Gao Aiping;Huang Jianfeng;Dang Zhiguo;Luo Ruixiong;Chen Yeyuan;Institute of Tropical Agriculture and Forestry, Hainan University;Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern China, Ministry of Agriculture, Tropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agricultural Sciences;National Center for Tropical Fruit Tree Variety Improvement;  
Red chlorophyll catabolite reductase(RCCR) is the key enzyme of chlorophyll degradation, it can decompose the red chl catabolit which is produced during the degradation of chlorophyll into primary fluorescent chl catabolite. In this study, degenerate primers were designed according to RCCR partial gene sequences by transcriptome sequencing to clone RCCR cDNA sequence with 3' and 5' RACE-PCR, three different colored mango varieties(red‘Guifei'Variety, yellow‘Jinhuang'and green‘Guiqi'varieties) were selected to clone of amplified full-length cDNA sequence of RCCR. It was found that only a few bases differences in the cDNA sequences of three different colored mango varieties. Take the yellow ‘Jinhuang'variety for example, the full-length cDNA of RCCR was 1 134 bp long. Its open reading frame(ORF) is 975 bp, encoding a protein of 324 amino acids with a predicted molecular weight of 36.21 k D. The isoelectric point of the predicted protein was 5.13. Three different protein structre was obtained by biological software prediction. It was found that the gene encoding for the protein had closerelationship with orange, Citrus clementina by other plants through phylogenetic analysis. In recent years,virus-induced gene silencing(VIGS) was a powerful tool for the study of gene function in plants. In order to investigate the role of RCCR gene in chlorophyll degradation, this study successfully constructed RCCR-p TRV2 gene silencing vector, to provides a theoretical basis for study the mechanism of RCCR gene in mango fruit ripening and pericarp coloration for the future.
【Fund】: 国家自然科学基金(31471850);; 农业部热带作物种质资源保护(15RZZY-07);; 948项目(2011-G13);; 非营利性科研机构改革启动经费(CATAS PZS-201225;CATAS-TCGRI;1630032013003);; 2017年度海南省研究生创新科研课题(Hys2017-74)共同资助
【CateGory Index】: Q943.2;S667.7
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