Cloning and Expression Analysis of OsNADPH1 Gene from Rice in Drought Stress Response

专利名称：Cloning, expression and use of acidphospholipases发明人：Khanh Quoc Nguyen,Kornelia Titze,Tatiana Schwarz,Silvia Paladino,VolkerMarschner,Patrick Lorenz申请号：US13502945申请日：20101027公开号：US09045713B2公开日：20150602专利内容由知识产权出版社提供摘要：The invention relates to a DNA sequence, which codes for a polypeptide having phospholipase activity essentially without lipase activity, characterized in that the DNA sequence is selected from a) DNA sequences that comprise a nucleotide sequence according to SEQ ID NO: 1, b) DNA sequences that comprise the coding sequence according to SEQ ID NO: 1, c) DNA sequences that code for the protein sequence according to SEQ ID NO: 2, d) DNA sequences that are coded for by the plasmid pPL3940-Topo2.5 with the restriction map according to FIG. , which is deposited under accession number DSM 22741, e) DNA sequences that hybridize under stringent conditions with one of the DNA sequences according to a), b), c) or d), f) DNA sequences that are related to the DNA sequences according to a), b), c), d) or e) due to the degeneration of the genetic code, and g) complementary strands to the sequences according to a) to f), wherein the DNA sequence is preferably derived from , and more preferably from , and a polypeptide having phospholipase activity essentially without lipase activity selected from a) a polypeptide which is coded for by the coding part of a DNA sequence as defined above,b) a polypeptide having the sequence according to SEQ ID NO: 2 or a sequence derived therefrom, which may be obtained by substitution, addition, deletion of one or more amino acid(s), c) a polypeptide having a sequence that has at least 83% identity with the amino acids 1 to 299 of SEQ ID NO: 2, d) a polypeptide which is coded for by a nucleic acid sequence which hybridizes under stringent conditions with (i) nucleotides 55 to 1106 of SEQ ID NO: 1, (ii) the cDNA sequence contained in nucleotides 55 to 1106 of SEQ ID NO: 1, (iii) a partial sequence of (i) or (ii) composed of at least 100 nucleotides, or (iv) a complementary strand of (i), (ii) or (iii), e) a variant of the polypeptide having SEQ ID NO: 2, comprising a substitution, deletion and/or insertion of one or more amino acid(s), f) allelic variants to amino acid sequences a) to e).申请人：Khanh Quoc Nguyen,Kornelia Titze,Tatiana Schwarz,Silvia Paladino,Volker Marschner,Patrick Lorenz地址：Reichelsheim DE,Muehltal DE,Erzhausen DE,Heppenheim DE,BickenbachDE,Lorsch DE国籍：DE,DE,DE,DE,DE,DE代理机构：Mintz Levin Cohn Ferris Glovsky and Popeo, P.C.代理人：Peter F. Corless更多信息请下载全文后查看。

专利名称：Cloning, expression and use of acidphospholipases发明人：Khanh Quoc Nguyen,Kornelia Titze,TatianaSchwarz,Silvia Paladino,VolkerMarschner,Patrick Lorenz申请号：US14692230申请日：20150421公开号：US09322003B2公开日：20160426专利内容由知识产权出版社提供专利附图：摘要：The invention relates to a DNA sequence, which codes for a polypeptide havingphospholipase activity essentially without lipase activity, characterized in that the DNA sequence is selected from a) DNA sequences that comprise a nucleotide sequence according to SEQ ID NO: 1, b) DNA sequences that comprise the coding sequence according to SEQ ID NO: 1, c) DNA sequences that code for the protein sequence according to SEQ ID NO: 2, d) DNA sequences that are coded for by the plasmid pPL3940-Topo2.5 with the restriction map according to FIG. , which is deposited under accession number DSM 22741, e) DNA sequences that hybridize under stringent conditions with one of the DNA sequences according to a), b), c) or d), f) DNA sequences that are related to the DNA sequences according to a), b), c), d) or e) due to the degeneration of the genetic code, and g) complementary strands to the sequences according to a) to f), wherein the DNA sequence is preferably derived from , and more preferably from , and a polypeptide having phospholipase activity essentially without lipase activity selected from a) a polypeptide which is coded for by the coding part of a DNA sequence as defined above, b) a polypeptide having the sequence according to SEQ ID NO: 2 or a sequence derived therefrom, which may be obtained by substitution, addition, deletion of one or more amino acid(s), c) a polypeptide having a sequence that has at least 83% identity with the amino acids 1 to 299 of SEQ ID NO: 2, d) a polypeptide which is coded for by a nucleic acid sequence which hybridizes under stringent conditions with (i) nucleotides 55 to 1106 of SEQ ID NO: 1, (ii) the cDNA sequence contained in nucleotides 55 to 1106 of SEQ ID NO: 1, (iii) a partial sequence of (i) or (ii) composed of at least 100 nucleotides, or (iv) a complementary strand of (i), (ii) or (iii), e) a variant of the polypeptide having SEQ ID NO: 2, comprising a substitution, deletion and/or insertion of one or more amino acid(s), f) allelic variants to amino acid sequences a) to e).申请人：AB Enzymes GMBH地址：Darmstadt DE国籍：DE代理机构：Mintz Levin Cohn Ferris Glovsky and Popeo, P.C.代理人：Peter F. Corless, Esq.,Daniel W. Clarke更多信息请下载全文后查看。

园艺学报，2016，43 (5)：975–982.Acta Horticulturae Sinicadoi：10.16420/j.issn.0513-353x.2016-0045；http：//www. ahs. ac. cn 975森林草莓独脚金内酯合成关键基因D27的克隆与表达分析赵晨晨1，范雅丽2，秦岭1,3，邢宇1,3，房克凤1,3，张卿1，曹庆芹2,3,*（1北京农学院植物科学与技术学院，农业应用新技术北京市重点实验室，北京 102206；2北京农学院生物科学与工程学院，北京 102206；3北京林果业生态环境功能提升协同创新中心，北京 102206）摘要：根据森林草莓中独角金内酯合成过程中的关键基因D27的序列信息设计引物，获得了D27基因的完整开放阅读框架。

采用MEGA5.0软件，对森林草莓与其他物种D27基因编码的氨基酸序列进行聚类分析，发现森林草莓D27基因与其他物种D27基因具有较高的同源性。

设置正常供磷、缺磷和在缺磷条件下接种丛枝状菌根真菌处理，利用电感耦合等离子发射光谱仪测定草莓植株的磷含量，结果表明缺磷条件下接种丛枝状菌根真菌，草莓植株中磷含量极显著提高，而缺磷处理的磷含量极显著降低。

实时荧光定量PCR结果表明，与正常供磷处理相比，缺磷和接种丛枝状菌根真菌处理条件下，D27基因的表达量分别上调7倍和11倍，这说明D27基因的表达既受到磷水平调控，又受到菌根形成的调控。

关键词：森林草莓；D27基因；聚类树；磷含量；表达分析中图分类号：S 668.4文献标志码：A 文章编号：0513-353X（2016）05-0975-08The Cloning and Expression Analysis of D27 Gene of Strigolactone Biosynthesis Pathway in Fragaria vescaZHAO Chen-chen1，FAN Ya-li2，QIN Ling1,3，XING Yu1,3，FANG Ke-feng1,3，ZHANG Qing1，and CAO Qing-qin2,3,*（1College of Plant Science and Technology，Beijing University of Agriculture，Beijing Key Laboratory for Agricultural Application and New Technique，Beijing 102206，China；2College of Biological Sciences and Engineering，Beijing University of Agriculture，Beijing 102206，China；3Beijing Collaborative Innovation Center for Eco-environmental Improvement with Forestry and Fruit Trees，Beijing 102206，China）Abstract：The complete coding sequence of D27 gene was cloned according to the sequence information of D27 in woodland strawberry genome，which is the key gene in biosynthesis pathway of phytohormone strigolactone. Phylogenetic tree was constructed by MEGA5.0 based on amino acid sequences of D27 genes derived from woodland strawberry and other plants. The results revealed that D27 gene in woodland strawberry possessed high sequence homology with other known D27 genes. Three different treatments were carried out in current study，which consisted of normal phosphorus application收稿日期：2016–03–25；修回日期：2016–05–05基金项目：北京市自然科学基金项目（6132005）；北京市属高等学校创新团队建设与教师职业发展计划项目（IDHT20140509）；国家自然科学基金项目（31201610）；科技创新服务能力建设—协同创新中心—林果业生态环境功能提升协同创新中心项目（PXM2016_014207_ 000038）* 通信作者Author for correspondence（E-mail：caoqingqin@）Zhao Chen-chen，Fan Ya-li，Qin Ling，Xing Yu，Fang Ke-feng，Zhang Qing，Cao Qing-qin.The cloning and expression analysis of D27 gene of strigolactone biosynthesis pathway in Fragaria vesca. 976Acta Horticulturae Sinica，2016，43 (5)：975–982.（control），phosphorus starvation and mycorrhiza fungi symbiosis under phosphorus starvation. Using plasma-atomic emission spectrometry（ICP-AES），the phosphorus content in Fragaria vesca under three treatments was detected. The results showed that the phosphorus content under mycorrhiza fungi symbiosis treatment was significantly higher，while the phosphorus content under phosphorus starvation was significantly lower when compared with control. Real-time quantitative PCR analysis showed that D27 was upregulated by 11 times in mycorrhizal roots and by 7 times in phosphorus starvation roots in comparison with that of the control roots，respectively. It revealed that the expression of D27 was regulated by both phosphorus level and mycorrhiza formation.Key words：Fragaria vesca；D27；phylogenetic analysis；phosphorus content；expression analysis独脚金内酯（Strigolactone）是近几年发现的一种植物激素（Umehara et al.，2008；王玫等，2014）。

热带作物学报2021, 42(5): 1209 1215Chinese Journal of Tropical Crops菠萝MYB基因AcoMYB1的克隆与表达分析李穆1，蔡元保1，杨祥燕1*，黄思婕1，李季东1，谭秦亮1，邱文武2，方位宽2 1. 广西农业科学院广西亚热带作物研究所，广西南宁 530001；2. 广西农业科学院园艺研究所，广西南宁 530007摘要：根据菠萝转录组的测序结果克隆到1个MYB转录因子基因，命名为AcoMYB1，GenBank登录号为XM_020230319。

该基因cDNA全长1221 bp，开放阅读框（ORF）为747 bp，编码一个含有248个氨基酸的蛋白。

序列分析表明，AcoMYB1氨基酸序列N端具有2个保守的SANT结构域，属于R2R3类MYB转录因子。

生物信息学分析表明，AcoMYB1是不稳定的亲水蛋白，不具有跨膜结构和信号肽，可能定位于细胞质，二级结构以α-螺旋和无规则卷曲为主。

实时荧光定量PCR分析表明，AcoMYB1在菠萝干旱、低温逆境胁迫处理下受诱导表达，整体上表现出“先升后降”的趋势；在早熟品种和晚熟品种的果实发育过程中也被诱导表达，表现为“升-降-升”的趋势，特别是在果实发育早期和果实成熟后期受诱导表达的强度较为突出。

由此推测AcoMYB1作为正调控因子参与菠萝冷害、干旱逆境胁迫的响应过程，并在菠萝果实早期发育及后期成熟发挥调控作用。

关键词：菠萝；MYB基因；基因表达；果实发育；逆境胁迫中图分类号：S668.3 文献标识码：ACloning and Expression Analysis of MYB Gene AcoMYB1 in Pineapple (Ananas comosus)LI Mu1, CAI Yuanbao1, YANG Xiangyan1*, HUANG Sijie1, LI Jidong1, TAN Qinliang1, QIU Wenwu2, FANG Weikuan21. Guangxi Subtropical Crops Research Institute, Guangxi Academy of Agricultural Sciences, Nanning, Guangxi 530001, China;2. Horticultural Research Institute, Guangxi Academy of Agricultural Sciences, Nanning, Guangxi 530007, ChinaAbstract: A MYB transcription factor gene, belonging to R2R3, was cloned by the results of the transcriptome se-quencing of pineapple (Ananas comosus), which was named AcoMYB1 and the GenBank accession number was XM_020230319. The full length cDNA and coding region (ORF) was 1221 bp and 747 bp, encoding 248 amino acids with two conserved SANT domains at N-terminal. By bioinformatics analysis, AcoMYB1 was an unstable hydrophilic protein without transmembrane structure and signal peptide, which may be located in the cytoplasm, and the secondary structure mainly had alpha helixes and random coils. Real-time fluorescence quantitative PCR analysis showed that AcoMYB1 was induced by drought and low temperature stress, and showed a trend of “up first then down”, and the ex-pression was also induced in the fruit development of early and late maturing varieties, which showed a tendency of “up-down-up”, especially in the early stage of fruit development and the later stage of fruit maturation. Therefore, it is speculated that AcoMYB1 as a positive regulatory factor is involved in the response process of chilling injury and drought stress, and plays a regulatory role in the early fruit development and late fruit maturation of pineapple (Ananas comosus).Keywords: Ananas comosus; MYB genes; gene expression; fruit development; adversity stressDOI: 10.3969/j.issn.1000-2561.2021.05.002收稿日期 2020-06-30；修回日期 2020-08-15基金项目 广西自然科学基金项目（No. 2019GXNSFAA245010）；广西农业科学院基本科研业务专项（桂农科2020YM55）；广西直属公益性科研院所基本科研业务费专项（桂热研201903）。