摘要
氨基酸通透酶(amino acid permease, AAP)参与植物氨基酸的转运、吸收、氮代谢等途径。为了解橄榄果实AAP基因在不同品种果实成熟发育过程的表达模式,本研究以呈味差异显著的两个品种普通橄榄‘长营’和清橄榄‘梅埔2号’为试验材料,通过RT-PCR方法克隆了3个APP基因的cDNA序列全长,并对其进行了生物学信息分析与亚细胞定位分析,同时利用实时荧光定量PCR技术分析了两个不同品种果实在不同发育时期的CaAAPs表达情况。结果表明:3个基因的序列长度分别为1 254 bp (CaAAP1)、405 bp (CaAAP2)、633 bp (CaAAP4),其编码的蛋白均为不稳定疏水性蛋白,分别含有7、1、5个跨膜区,保守基序差异较大;亚细胞定位结果表明,CaAAP1、CaAAP2主要在细胞质膜上表达。荧光定量结果表明随着果实的发育,CaAAP1、CaAAP2和CaAAP4在‘长营’果实的表达量均呈显著上升趋势,在‘梅埔2号’的表达量变化趋势较平缓,且在170 d时显著下降,两个品种CaAAPs家族基因表达量差异大,表明CaAAP1、CaAAP2、CaAAP4基因可能对橄榄果实游离氨基酸转运和吸收起着重要作用。该研究结果为进一步研究CaAAPs在橄榄果实氨基酸合成代谢中的功能提供理论依据。
Amino acid permease(AAP)is involved in the transportation,absorption,and nitrogen metabolism of plant amino acids.In order to understand the expression pattern of AAP gene in Canarium album(Lour.)Raeusch fruit during the ripening and development of different varieties,this study used two varieties Chinese olive with significant differences in taste,ordinary olives‘Changying'and sweet olive Meipu2'as experimental materials,and passed RT-PCR method cloned the full-length cDNA sequence of 3 APP genes,and carried out biological information analysis and subcellular localization analysis on them.At the same time,real-time fluorescent quantitative PCR technology was used to analyze the CaAAPs expression of two different varieties of fruits at different developmental stages.The results showed that the sequence lengths of the three genes were 1254 bp(CaAAPI),405 bp(CaAA P2),and 633 bp(CaAA P4),respectively,and the proteins encoded by them were all unstable hydrophobic proteins.Each of the three genes contained 7,1,and 5 transmembrane regions.In the membrane region,the conservative motifs were quite different.The results of subcellular localization indicated that CaAAPl and CaAA P2 were mainly expressed on the plasma membrane.Fluorescence quantitative results showed that with the development of the fruit,the expression of CaAA P1,CaAA P2 and CaAA P4 in the‘Changying'fruits showed a significant upward trend,and the expression changes in the‘Meipu2'showed a relatively flat trend,and at 170 d significant decline.The two varieties of CaAAPs family gene expression levels were quite different,indicating that CaAAP1,CaAA P2,CaAA P4 genes may play an important role in the free amino acid transport and absorption of olive fruits.The results of this study provide a theoretical basis for further research on the function of CaAA Ps in the amino acid synthesis and metabolism of olive fruits.
作者
邱娜娜
谢倩
叶清华
王威
樊玥
罗培润
陈清西
Qiu Nana;Xie Qian;Ye Qinghua;Wang Wei;Fan Yue;Luo Peirun;Chen Qingxi(College of Horticulture,Fujian Agriculture and Forestry University,Fuzhou,350002)
出处
《分子植物育种》
CAS
北大核心
2023年第8期2517-2526,共10页
Molecular Plant Breeding
基金
福建农林大学横向科技创新基金项目(102-KHF200005)和福建农林大学科技创新专项基金项目(CXZX-2018087,CXZX2017186)共同资助。
关键词
橄榄
AAPs
克隆
亚细胞定位
表达分析
Canarium album(Lour.)Raeusch
AAPs
Cloning
Subcellular localization
Expression analysis
