Xiuxiu Zhao, Yangen Fan, Yueyue Tian, Hanyue Wang, Lixia Zhang, Min Li
{"title":"茶树叶绿体靶向铁氧化还原蛋白nadp +氧化还原酶基因的克隆与表达分析Huangjinya","authors":"Xiuxiu Zhao, Yangen Fan, Yueyue Tian, Hanyue Wang, Lixia Zhang, Min Li","doi":"10.5376/jtsr.2020.10.0001","DOIUrl":null,"url":null,"abstract":"The chloroplast-targeted ferredoxin-NADP+ oxidoreductase (LFNR) is a hydrophilic flavin protein located at the end of the photosynthesis electron delivery chain in higher plants. It’s a hub for linking photosynthesis electronic transportation with chloroplast redox metabolism. In order to explore the relationship between LFNR gene and yellowing of ‘Huangjinya’controlled by light, we took separately one bud two leaves of ‘Huangjinya’ ‘Shuchazao’(CK) as the material to clone CsLFNR gene and then we obtained the sequence with a similarity of 100%, named CsLFNR1.1 (GenBank accession no. MT311318). Throw bioinformatics analysis, we know its cDNA gene length is 1 095 bp, coding 364 amino acids, The protein molecular weight is 40.623KDa, the theoretical isoelectric point is 8.86.CsLFNR1.1 is a alkaline protein without transmembrane structure and signal peptide, it has a chloroplast transport peptide (cTP), the secondary structure contains 23.35% alpha helix, 5.49% beta-corner, 28.85% extension chain and 42.31% irregular curling, It locates in the chloroplast. Through the Nucleotide BLAST, Protein BLAST and DNAMAN software, we found that CsLFNR1.1 and its translated amino acid sequences had 4 bases and 3 amino acid differences with NCBI ‘Shuchazao’ Gene CsLFNR1 (XM_028233617.1) and Protein (XP_028089418.1), the similarity reached 99.63% and 99.18%. CsLFNR1.1 and CsLFNR1 have small differences in the physical and chemical properties and secondary structure, the tertiary structure prediction template of them is consistent. In addition, they have the same conserved domain and active site. We took different shade ‘Huangjinya’ blades as the material for qRT-PCR, the result showed that the expression of CsLFNR1.1 gene response to the light intensity, its expression increased with the increase of light intensity. This study provides a theoretical basis and scientific basis for further exploring the role of CsLFNR1.1 gene in the light regulation process of the new shoots and leaves of ‘Huangjinya’.","PeriodicalId":17156,"journal":{"name":"茶叶科学","volume":"360 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2020-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Cloning and Expression Analysis of Chloroplast-targeted Ferredoxin-NADP+ Oxidoreductase Gene of Tea Plant (Camellia sinensis) cv.Huangjinya\",\"authors\":\"Xiuxiu Zhao, Yangen Fan, Yueyue Tian, Hanyue Wang, Lixia Zhang, Min Li\",\"doi\":\"10.5376/jtsr.2020.10.0001\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"The chloroplast-targeted ferredoxin-NADP+ oxidoreductase (LFNR) is a hydrophilic flavin protein located at the end of the photosynthesis electron delivery chain in higher plants. It’s a hub for linking photosynthesis electronic transportation with chloroplast redox metabolism. In order to explore the relationship between LFNR gene and yellowing of ‘Huangjinya’controlled by light, we took separately one bud two leaves of ‘Huangjinya’ ‘Shuchazao’(CK) as the material to clone CsLFNR gene and then we obtained the sequence with a similarity of 100%, named CsLFNR1.1 (GenBank accession no. MT311318). Throw bioinformatics analysis, we know its cDNA gene length is 1 095 bp, coding 364 amino acids, The protein molecular weight is 40.623KDa, the theoretical isoelectric point is 8.86.CsLFNR1.1 is a alkaline protein without transmembrane structure and signal peptide, it has a chloroplast transport peptide (cTP), the secondary structure contains 23.35% alpha helix, 5.49% beta-corner, 28.85% extension chain and 42.31% irregular curling, It locates in the chloroplast. Through the Nucleotide BLAST, Protein BLAST and DNAMAN software, we found that CsLFNR1.1 and its translated amino acid sequences had 4 bases and 3 amino acid differences with NCBI ‘Shuchazao’ Gene CsLFNR1 (XM_028233617.1) and Protein (XP_028089418.1), the similarity reached 99.63% and 99.18%. CsLFNR1.1 and CsLFNR1 have small differences in the physical and chemical properties and secondary structure, the tertiary structure prediction template of them is consistent. In addition, they have the same conserved domain and active site. We took different shade ‘Huangjinya’ blades as the material for qRT-PCR, the result showed that the expression of CsLFNR1.1 gene response to the light intensity, its expression increased with the increase of light intensity. This study provides a theoretical basis and scientific basis for further exploring the role of CsLFNR1.1 gene in the light regulation process of the new shoots and leaves of ‘Huangjinya’.\",\"PeriodicalId\":17156,\"journal\":{\"name\":\"茶叶科学\",\"volume\":\"360 1\",\"pages\":\"\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2020-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"茶叶科学\",\"FirstCategoryId\":\"1087\",\"ListUrlMain\":\"https://doi.org/10.5376/jtsr.2020.10.0001\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"茶叶科学","FirstCategoryId":"1087","ListUrlMain":"https://doi.org/10.5376/jtsr.2020.10.0001","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Cloning and Expression Analysis of Chloroplast-targeted Ferredoxin-NADP+ Oxidoreductase Gene of Tea Plant (Camellia sinensis) cv.Huangjinya
The chloroplast-targeted ferredoxin-NADP+ oxidoreductase (LFNR) is a hydrophilic flavin protein located at the end of the photosynthesis electron delivery chain in higher plants. It’s a hub for linking photosynthesis electronic transportation with chloroplast redox metabolism. In order to explore the relationship between LFNR gene and yellowing of ‘Huangjinya’controlled by light, we took separately one bud two leaves of ‘Huangjinya’ ‘Shuchazao’(CK) as the material to clone CsLFNR gene and then we obtained the sequence with a similarity of 100%, named CsLFNR1.1 (GenBank accession no. MT311318). Throw bioinformatics analysis, we know its cDNA gene length is 1 095 bp, coding 364 amino acids, The protein molecular weight is 40.623KDa, the theoretical isoelectric point is 8.86.CsLFNR1.1 is a alkaline protein without transmembrane structure and signal peptide, it has a chloroplast transport peptide (cTP), the secondary structure contains 23.35% alpha helix, 5.49% beta-corner, 28.85% extension chain and 42.31% irregular curling, It locates in the chloroplast. Through the Nucleotide BLAST, Protein BLAST and DNAMAN software, we found that CsLFNR1.1 and its translated amino acid sequences had 4 bases and 3 amino acid differences with NCBI ‘Shuchazao’ Gene CsLFNR1 (XM_028233617.1) and Protein (XP_028089418.1), the similarity reached 99.63% and 99.18%. CsLFNR1.1 and CsLFNR1 have small differences in the physical and chemical properties and secondary structure, the tertiary structure prediction template of them is consistent. In addition, they have the same conserved domain and active site. We took different shade ‘Huangjinya’ blades as the material for qRT-PCR, the result showed that the expression of CsLFNR1.1 gene response to the light intensity, its expression increased with the increase of light intensity. This study provides a theoretical basis and scientific basis for further exploring the role of CsLFNR1.1 gene in the light regulation process of the new shoots and leaves of ‘Huangjinya’.
期刊介绍:
The Journal of Tea Science was established in August 1964, approved by the Publicity Department, CCCPC. Its title was inscribed by Zhu De, the chairman of CCCPC. It was discontinued during the Cultural Revolution in 1966, and it was reissued in August 1984, approved by the State Scientific and Technological Commission.Academicians Chen Zongmao and Liu Zhonghuaof the Chinese Academy of Engineering served as the directors of the editorial board. The Journal of Tea Science is managed by the China Association for Science and Technology,sponsored by the China Tea Science Society and the Tea Research Institute of the Chinese Academy of Agricultural Sciences, and edited and published by the editorial office of the Journal of Tea Science. It is the only one of Chinese core journals in the field of tea science that is included in the core library of the Chinese Science Citation Database.Its Domestic Unified Serial Number is CN 33-1115/S, its International Standard Serial Number is ISSN 1000-369X and its International publication name code is CODEN-CHKEF4. At present, the Journal of Tea Science is a bimonthly publication, published in the middle of the month, with a book size of 16.