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Cloning and expression analysis of phytoene desaturase and e-carotene desaturase genes in Carica papaya

机译:番木瓜中八氢番茄红素脱氢酶和e-胡萝卜素脱氢酶基因的克隆与表达分析

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摘要

The fruit flesh color of papaya is an important nutritional quality trait and is due to the accumulation of carotenoid. To elucidate the carotenoid biosynthesis pathway in Carica papaya, the phytoene desaturase (PDS) and the e-carotene desaturase (ZDS) genes were isolated from papaya (named CpPDS and CpZDS) using the rapid amplification of cDNA ends (RACE) approach, and their expression levels were investigated in red- and yellow-fleshed papaya varieties. CpPDS contains a 1749 bp open reading frame coding for 583 amino acids, while CpZDS contains a 1716 bp open reading frame coding for 572 amino acids. The deduced CpPDS and CpZDS proteins contain a conserved dinucleotide-binding site at the N-terminus and a carotenoid-binding domain at the C-terminus. Papaya genome sequence analysis revealed that CpPDS and CpZDS are single copy; the CpPDS was mapped to papaya chromosome LG6, and the CpZDS was mapped to chromosome LG3. Quantitative PCR showed that both CpPDS and CpZDS were expressed in all tissues examined with the highest expression in maturing fruits, and that the expression of CpPDS and CpZDS were higher in red-fleshed fruits than in yellow-fleshed fruits. These results indicated that the differential accumulation of carotenoids in red- and yellow-fleshed papaya varieties might be partly explained by the transcriptional level of CpPDS and CpZDS.
机译:木瓜的果肉颜色是重要的营养品质性状,归因于类胡萝卜素的积累。为了阐明番木瓜中的类胡萝卜素生物合成途径,使用cDNA末端快速扩增(RACE)方法从番木瓜中分离出了番茄红素去饱和酶(PDS)和电子胡萝卜素去饱和酶(ZDS)基因(分别称为CpPDS和CpZDS)。研究了红肉和黄肉木瓜品种的表达水平。 CpPDS包含一个1749 bp的开放阅读框,编码583个氨基酸,而CpZDS包含1716 bp的开放阅读框,编码572个氨基酸。推导的CpPDS和CpZDS蛋白在N端含有一个保守的二核苷酸结合位点,在C端含有一个类胡萝卜素结合域。木瓜基因组序列分析表明,CpPDS和CpZDS是单拷贝。 CpPDS定位于木瓜染色体LG6,CpZDS定位于染色体LG3。定量PCR显示CpPDS和CpZDS在所有检测的组织中均表达,在成熟果实中表达最高,并且在红肉果实中CpPDS和CpZDS的表达高于黄肉果实。这些结果表明,在红色和黄色肉木瓜品种中类胡萝卜素的差异积累可能部分由CpPDS和CpZDS的转录水平解释。

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