大豆<i>GmNAC011</i>基因的分离及对异黄酮合酶基因<i>GmIFS2</i>的调控分析

doi:10.7668/hbnxb.2015.05.004

华北农学报 ›› 2015, Vol. 30 ›› Issue (5): 25-29. doi: 10.7668/hbnxb.2015.05.004

所属专题： 油料作物；生物技术；

大豆GmNAC011基因的分离及对异黄酮合酶基因GmIFS2的调控分析

刘晓庆, 陈华涛, 张红梅, 袁星星, 崔晓艳, 顾和平, 陈新

江苏省农业科学院蔬菜研究所, 江苏南京 210014

收稿日期:2015-07-22 出版日期:2015-10-28
通讯作者: 陈新(1973-),男,江苏射阳人,研究员,博士,主要从事豆类育种研究。
作者简介:刘晓庆(1982-),女,江苏如皋人,副研究员,博士,主要从事植物抗逆机制研究。
基金资助:
江苏省自然科学基金项目(BK20130727);江苏省农业科技自主创新资金项目[CX(12)5021];江苏省大豆科技支撑项目(BE2013350);食用豆现代产业技术体系项目(CARS-09)

Isolation and Regulation Analysis of GmNAC011 Gene to Isoflavone Synthase Gene GmIFS2

LIU Xiao-qing, CHEN Hua-tao, ZHANG Hong-mei, YUAN Xing-xing, CUI Xiao-yan, GU He-ping, CHEN Xin

Institute of Vegetable Crops, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China

Received:2015-07-22 Published:2015-10-28

摘要/Abstract

摘要： 通过对不同异黄酮含量品种大豆的转录组测序分析,发现 GmNAC011 基因在不同品种间的表达量存在显著差异。为了研究该基因的功能,根据大豆基因组序列设计引物,从大豆根系中克隆到 GmNAC011 基因完整的开放性阅读框(ORF)(Opening reading frame)序列,通过生物信息学对该序列进行分析,结果显示:该基因编码268个氨基酸,蛋白质分子量为31 kDa,理论等电点为8.3。进化树分析发现该蛋白与GmNAC2亲缘关系较近,属于NAC转录因子ATAF1亚家族。通过RT-PCR的方法分析了 GmNAC011 基因与异黄酮合酶基因 GmIFS2 在不同组织中的表达模式,结果显示 GmNAC011 与 GmIFS2 呈现相似的表达趋势,均为在花中表达量相对较低,而在根、茎、叶和荚中的表达量较高;通过酵母单杂交实验发现 GmNAC011 可以与 GmIFS2 基因启动子中的“CGTG”基序结合,这些结果说明了 GmNAC011 基因参与调控 GmIFS2 基因的表达,进而可以调控异黄酮的合成。

关键词: 大豆, 转录因子, GmNAC011, 异黄酮合酶基因

Abstract: Based on the previous results of transcriptome sequencing,the expression of GmNAC011 was different in soybean cultivars that differed in isoflavonoid accumulation,thus we selected this gene for further analysis.Primers were designed according to the soybean genome sequences.The whole opening reading frame of GmNAC011 was isolated from soybean roots,encoding 268 amino acids with a calculated molecular mass of 31 kDa and a theoretical pI of 8.3. Phylogenetic analysis showed that GmNAC011 was close to GmNAC2,belonging to ATAF1-like NAC transcription factor subgroup.We investigated the expression pattern of GmNAC011 and GmIFS2 in various tissues(root,stem,leaf,flower and pod),the expression model of GmNAC011 was similar with GmIFS2. The transcript levels of GmNAC011 and GmIFS2 were found to be higher in roots,stems,leaves and pods than flowers.And we found that GmNAC011 could binding to the motif "CGTG" in the promoter of GmIFS2 by yeast one-hybrid.All results showed that GmNAC011 participate in regulating the expression of GmIFS2,subsequently the synthesis of isoflavonoid.

Key words: Soybean, Transcription factor, GmNAC011, GmIFS2

中图分类号:

S565.01
Q78

刘晓庆, 陈华涛, 张红梅, 袁星星, 崔晓艳, 顾和平, 陈新. 大豆GmNAC011基因的分离及对异黄酮合酶基因GmIFS2的调控分析[J]. 华北农学报, 2015, 30(5): 25-29. doi: 10.7668/hbnxb.2015.05.004.

LIU Xiao-qing, CHEN Hua-tao, ZHANG Hong-mei, YUAN Xing-xing, CUI Xiao-yan, GU He-ping, CHEN Xin. Isolation and Regulation Analysis of GmNAC011 Gene to Isoflavone Synthase Gene GmIFS2[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(5): 25-29. doi: 10.7668/hbnxb.2015.05.004.

http://www.hbnxb.net/CN/Y2015/V30/I5/25

参考文献

[1] Duffy C,Perez K,Partridge A.Implications of phytoestrogen intake for breast cancer[J].CA-A Cancer Journal for Clinicians,2007,57(5):260-277.
[2] Nestel P.Isoflavones:their effects on cardiovascular risk and function[J].Current Opinion in Lipidology,2003,14(1):3-8.
[3] Morris P F,Bone E,Tyler B M.Chemotropic and contact responses of Phytophthora sojae hyphae to soybean isoflavonoids and artificial substrates[J].Plant Physiology,1998,117(4):1171-1178.
[4] Pueppke J L.The genetics and biochemical basis for nodulation of legumes by rhizobia[J].Critical Reviews in Biotechnology,1996,16(1):1-51.
[5] Akashi T,Aoki T,Ayabe S.Cloning and functional expression of a cytochrome P450 cDNA encoding 2-hydroxyisoflavanone synthase involved in biosynthesis of the isoflavonoid skeleton in licorice[J].Plant Physiology,1999,121(3):821-828.
[6] Steele C L,Gijzen M,Qutob D,et al.Molecular characterization of the enzyme catalyzing the aryl migration reaction of isoflavonoid biosynthesis in soybean[J].Archives of Biochemistry and Biophysics,1999,367(1):146-150.
[7] Jung W,Yu O,Lau S C,et al.Identification and expression of isoflavone synthase,the key enzyme for biosynthesis of isoflavones in legumes[J].Nat Biotech,2000,18(2):208-212.
[8] Liu X Q,Yuan L L,Xu L,et al.Over-expression of GmMYB39 leads to an inhibition of the isoflavonoid biosynthesis in soybean(Glycine max L)[J].Plant Biotechnol Rep,2013,7(4):445-455.
[9] Aida M,Ishida T,Fukaki H,et al.Genes involved in organ separation in Arabidopsis:an analysis of the cup-shaped cotyledon mutant[J].The Plant Cell,1997,9(6):841-857.
[10] Xie Q,Frugis G,Colgan D,et al.Arabidopsis NAC1 transduces auxin signal downstream of TIR1to promote lateral root development[J].Genes & Development,2000,14(23):3024-3036.
[11] Duval M,Hsieh T F,Kim S Y,et al.Molecular characterization of AtNAM:a member of the Arabidopsis NAC domain superfamily[J].Plant Molecular Biology,2002,50(2):237-248.
[12] Ernst H A,Olsen A N,Larsen S,et al.Structure of the conserved domain of ANAC,a member of the NAC family of transcription factors[J].EMBO Reports,2004,5(3):297-303.
[13] Ooka H,Satoh K,Doi K,et al.Comprehensive analysis of NAC family genes in Oryza sativa and Arabidopsis thaliana[J].DNA Research,2003,10(6):239-247.
[14] Jin H X,Huang F,Cheng H,et al.Overexpression of the GmNAC2 gene,an NAC transcription factor,reduces abiotic stress tolerance in Tobacco[J].Plant Molecular Biology Reporter,2013,31(2):435-442.
[15] Olsen A N,Ernst H A,Leggio L L,et al.NAC transcription factors:structurally distinct,functionally diverse[J].Trends in Plant Science,2005,10(2):79-87.
[16] 才华,朱延明,李勇,等.野生大豆转录因子 GsNAC20 基因的分离及胁迫耐性分析[J].作物学报,2011,37(8):1351-1359.
[17] Meng Q,Zhang C,Gai J,et al.Molecular cloning,sequence characterization and tissue-specific expression of six NAC-like genes in soybean(Glycine max L.Merr.)[J].Journal of Plant Physiology,2007,164(8):1002-1012.
[18] Hegedus D,Yu M,Baldwin D,et al.Molecular characterization of Brassica napus NAC domain transcriptional activators induced in response to biotic and abiotic stress[J].Plant Molecular Biology,2003,53(3):383-397.
[19] Fujita M,Fujita Y,Maruyama K,et al.A dehydration-induced NAC protein,RD26,is involved in a novel ABA-dependent stress-signaling pathway[J].Plant Journal,2004,39(6):863-876.
[20] Tran L S,Nakashima K,Sakuma Y,et al.Isolation and functional analysis of Arabidopsis stress-inducible NAC transcription factors that bind to a drought-responsive cis-element in the early responsive to dehydration stress 1 promoter[J].Plant Cell,2004,16(9):2481-2498.

选择文件类型/文献管理软件名称

选择包含的内容

大豆GmNAC011基因的分离及对异黄酮合酶基因GmIFS2的调控分析

Isolation and Regulation Analysis of GmNAC011 Gene to Isoflavone Synthase Gene GmIFS2

PDF

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	邹晓悦, 刘佳, 李志勇, 马继芳, 王永芳, 全建章, 刘磊, 白辉, 董志平. *谷子SibHLH19基因的克隆及表达分析*[J]. 华北农学报, 2022, 37(5): 16-24.
[2]	贾建平, 李金鸿, 于敬文, 余曦玥, 彭德良, 李惠霞, 黄文坤. 氮肥对控制大豆孢囊线虫病危害的影响[J]. 华北农学报, 2022, 37(4): 198-205.
[3]	张斌. *大豆GmPP2C89基因在盐胁迫中的功能分析*[J]. 华北农学报, 2022, 37(4): 20-27.
[4]	李艳燕, 赵彩桐, 齐阳阳, 刘明, 张书瑞, 刘志华, 李文滨, 姜振峰. 大豆主茎木质素积累规律分析[J]. 华北农学报, 2022, 37(3): 77-85.
[5]	张晋玉, 徐新娟, 晁毛妮, 张晓红, 吴向远, 高际涛, 黄中文. *大豆GmZAT12基因的克隆和表达分析*[J]. 华北农学报, 2022, 37(3): 1-7.
[6]	吉颖, 孙枫, 吴淑华, 李硕, 涂丽琴, 高丹娜, 崔晓艳, 陈新, 季英华, 郭青云. 江苏大豆上分离的豇豆轻斑驳病毒基因组序列克隆及特征分析[J]. 华北农学报, 2022, 37(3): 200-205.
[7]	王晓丽, 王敏, 岳爱琴, 郭数进, 王鹏, 王利祥, 杨婷婷, 张海生, 张永坡, 高春艳, 张武霞, 牛景萍, 杜维俊, 赵晋忠. 氮素营养和根瘤菌接种对大豆结瘤固氮和生长的影响[J]. 华北农学报, 2022, 37(1): 95-102.
[8]	李曼, 史晓蕾, 邸锐, 刘志芳, 孟庆民, 付才, 杨春燕, 王冬梅, 张孟臣, 张洁, 闫龙. 大豆长片段插入/缺失标记的开发与应用[J]. 华北农学报, 2022, 37(1): 18-26.
[9]	吴红红, 段学粉, 郭仰东, 张喜春. *转录因子SlMYB-related 2对番茄耐冷性的影响*[J]. 华北农学报, 2022, 37(1): 35-41.
[10]	韦锦坚, 覃潇敏, 农玉琴, 骆妍妃, 陆金梅, 陈远权, 韦持章. 茶与大豆间作对土壤微生物群落代谢功能多样性的影响[J]. 华北农学报, 2021, 36(S1): 289-296.
[11]	王大刚, 陈圣男, 黄志平, 李杰坤, 吴倩, 胡国玉, 王维虎, 杨永庆. 大豆新品系抗SMV鉴定及分析[J]. 华北农学报, 2021, 36(S1): 326-332.
[12]	高倩, 冯燕, 杨雅华, 赵青松, 雷雅坤, 刘兵强, 张孟臣, 史晓蕾, 杨春燕. 基于全基因组关联分析挖掘野生大豆蛋白含量QTL[J]. 华北农学报, 2021, 36(S1): 23-30.
[13]	雷其冬, 孙旭东, 徐慧妮. 转录因子TCP4参与植物生长发育和抗逆调节研究进展[J]. 华北农学报, 2021, 36(S1): 210-214.
[14]	翟玲侠, 于崧, 侯玉龙, 秦猛, 朱雪天, 王小琴, 于立河. 芸豆盐碱响应NAC转录因子的分子特性分析[J]. 华北农学报, 2021, 36(6): 19-27.
[15]	范会芬, 孙天杰, 苏伟华, 肖付明, 张洁, 王冬梅. 大豆GmWRKY50的抗体制备及其蛋白水平表达分析[J]. 华北农学报, 2021, 36(6): 28-34.

模态框（Modal）标题

选择文件类型/文献管理软件名称

选择包含的内容

大豆GmNAC011基因的分离及对异黄酮合酶基因GmIFS2的调控分析

Isolation and Regulation Analysis of GmNAC011 Gene to Isoflavone Synthase Gene GmIFS2

PDF

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价