茶叶科学
茶葉科學
다협과학
2015年
1期
55-63
,共9页
肖瑶%周天山%李佼%张佳欣%余有本
肖瑤%週天山%李佼%張佳訢%餘有本
초요%주천산%리교%장가흔%여유본
茶树%GDP-D-甘露糖焦磷酸化酶%基因克隆%表达分析
茶樹%GDP-D-甘露糖焦燐痠化酶%基因剋隆%錶達分析
다수%GDP-D-감로당초린산화매%기인극륭%표체분석
Camellia sinensis%GDP-D-mannosepyrophosphorylase(GMP)%geneclonging%expression analysis
抗坏血酸(Vc)是重要的抗氧化剂,在茶树体内的各种生理代谢及抵御非生物胁迫中起重要作用,Vc 含量关系到绿茶品质优劣。GDP-D-甘露糖焦磷酸化酶(GMP)是茶树 Vc 生物合成途径中的关键酶之一。本研究通过 RACE-PCR 方法从茶树中克隆了 GMP cDNA 全长序列,共计1510 bp,其中包含1086 bp 的开放阅读框(ORF),编码361个氨基酸,推测蛋白分子量为39.599 kDa。BLAST 分析表明,茶树 GMP 基因氨基酸序列与猕猴桃的亲缘性最高,达到96%。实时定量 PCR 分析表明,茶树新梢芽下第三叶中 GMP 基因表达量最高,嫩茎中最低,不同品种茶树新梢叶片中表达量存在明显差异。高温胁迫初期,GMP 基因表达量及抗坏血酸含量迅速升高,然后逐渐下降且均低于同期对照。
抗壞血痠(Vc)是重要的抗氧化劑,在茶樹體內的各種生理代謝及牴禦非生物脅迫中起重要作用,Vc 含量關繫到綠茶品質優劣。GDP-D-甘露糖焦燐痠化酶(GMP)是茶樹 Vc 生物閤成途徑中的關鍵酶之一。本研究通過 RACE-PCR 方法從茶樹中剋隆瞭 GMP cDNA 全長序列,共計1510 bp,其中包含1086 bp 的開放閱讀框(ORF),編碼361箇氨基痠,推測蛋白分子量為39.599 kDa。BLAST 分析錶明,茶樹 GMP 基因氨基痠序列與獼猴桃的親緣性最高,達到96%。實時定量 PCR 分析錶明,茶樹新梢芽下第三葉中 GMP 基因錶達量最高,嫩莖中最低,不同品種茶樹新梢葉片中錶達量存在明顯差異。高溫脅迫初期,GMP 基因錶達量及抗壞血痠含量迅速升高,然後逐漸下降且均低于同期對照。
항배혈산(Vc)시중요적항양화제,재다수체내적각충생리대사급저어비생물협박중기중요작용,Vc 함량관계도록다품질우렬。GDP-D-감로당초린산화매(GMP)시다수 Vc 생물합성도경중적관건매지일。본연구통과 RACE-PCR 방법종다수중극륭료 GMP cDNA 전장서렬,공계1510 bp,기중포함1086 bp 적개방열독광(ORF),편마361개안기산,추측단백분자량위39.599 kDa。BLAST 분석표명,다수 GMP 기인안기산서렬여미후도적친연성최고,체도96%。실시정량 PCR 분석표명,다수신소아하제삼협중 GMP 기인표체량최고,눈경중최저,불동품충다수신소협편중표체량존재명현차이。고온협박초기,GMP 기인표체량급항배혈산함량신속승고,연후축점하강차균저우동기대조。
Ascorbic acid(Vitamin C, Vc) is an important antioxidant in tea plant, playing important roles in metabolism and responses to abiotic stress, and tea qualityis positive correlation with the content ofVc.GDP-D-mannosepyrophosphorylase(GMP) is an important enzyme in the synthesis of ascorbic acid.The full-length cDNA sequence ofGMPgene was isolated from the shoots ofCamellia sinensisby RT-PCR and RACE. The entire GMPcDNAwas 1510bp, containing a 1086bp complete open reading frame which encoding a protein with 361 amino acidsand a calculated molecular weight of39.599kDa.Blastanalysis showedthatGMPgene in Camellia sinensiswas most closely toActinidiawith 96% amino acids similarity.Quantitative real-time PCR analysis showed that the expression levels ofGMPgene in the third leaves were highest while the stem is the lowest, and the different varieties also existed an obvious differences. High temperature stress stimulate the expression of GMPgene and the accumulation ofVcin incipient stage, then reduced rapidly.