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采用RT-PCR和RACE技术,从刺梨(Rosa roxburghii Tratt)果实中扩增出GDP-L–半乳糖磷酸酶(GDP-L-galactose pyrophosphatase,GGP)基因的全长cDNA序列,并分析其在不同组织和果实不同发育阶段的表达特点及其与AsA积累的关系。结果表明:刺梨GGP基因cDNA(RrGGP,GenBank登录号:HM998753)包含1个长度为1 341 bp的开放阅读框(ORF),编码445个氨基酸;其氨基酸序列与苹果、猕猴桃等的相似性均在80%以上。RT-PCR分析表明,该基因在刺梨不同器官中的表达差异明显:在果实中的表达量最强,叶中次之,而花和茎中只能检测到极微弱的表达。在果实不同发育时期都能检测到RrGGP的表达,在发育初期(花后50 d内)表达较弱,花后60~100 d呈高水平表达,而后随着果实成熟而下调。刺梨中RrGGP的表达特点与AsA积累高度一致,意味着该基因可能是果实发育过程中AsA合成的关键基因。
The full-length cDNA sequence of GDP-L-galactose pyrophosphatase (GGP) gene was amplified from the fruit of Rosa roxburghii Tratt by RT-PCR and RACE, Expression Characteristics of Different Tissues and Fruits at Different Developmental Stages and Their Relationship with AsA Accumulation. The results showed that the cDNA of RgGGP (GenBank Accession No. HM998753) contained a 1 341 bp open reading frame (ORF) encoding a protein of 445 amino acids. The amino acid sequence of the cDNA was similar to that of apple and kiwifruit Above 80%. RT-PCR analysis showed that the expression of this gene was significantly different in different organs of Rosa roxburghii. The expression was the strongest in the fruit, followed by the leaf, while only very weak expression was detected in the flower and stem. The expression of RrGGP was detected at different developmental stages of fruit, and was weakly expressed at the early stage of development (within 50 days after anthesis). The expression of RrGGP was highly expressed at 60 to 100 days after anthesis, and then decreased as the fruit matured. The expression of RrGGP in Rosa roxburghii was highly consistent with the accumulation of AsA, suggesting that this gene may be the key gene for AsA synthesis during fruit development.