[1]张古文,刘娜,冯志娟,等.大豆异黄酮还原酶基因的鉴定及生物信息学分析[J].大豆科学,2016,35(06):911-918.[doi:10.11861/j.issn.1000-9841.2016.06.0911]
　ZHANG Gu-wen,LIU Na,FENG Zhi-juan,et al.Identification and Bioinformatics Analysis of Isoflavone Reductase Genes in Soybean[J].Soybean Science,2016,35(06):911-918.[doi:10.11861/j.issn.1000-9841.2016.06.0911]
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大豆异黄酮还原酶基因的鉴定及生物信息学分析

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第35卷 期数: 2016年06期 页码: 911-918 栏目: 出版日期: 2016-11-20
Title:
Identification and Bioinformatics Analysis of Isoflavone Reductase Genes in Soybean
作者:
张古文; 刘娜; 冯志娟; 徐盛春; 胡齐赞; 龚亚明
（浙江省农业科学院 蔬菜研究所，浙江 杭州 310021）
Author(s):
ZHANG Gu-wen;  LIU Na;  FENG Zhi-juan;  XU Sheng-chun;  HU Qi-zan;  GONG Ya-ming
（Institute of vegetables, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China）
关键词:
大豆; 异黄酮; 异黄酮还原酶; 基因家族
Keywords:
Soybean;  Isoflavone;  Isoflavone reductase;  Gene family
DOI:
10.11861/j.issn.1000-9841.2016.06.0911
文献标志码:
A
摘要:
异黄酮是大豆生长过程中形成的一类次级代谢产物，是大豆主要的功能活性成分之一。异黄酮还原酶（isoflavone reductase，IFR）是异黄酮分解途径的关键酶之一，在调控异黄酮含量及成分方面起着重要作用。本研究基于大豆全基因组测序数据，利用生物信息学方法鉴定了大豆异黄酮还原酶基因家族成员，并对其基因的序列特征、结构特征和遗传多样性进行了分析。结果表明：大豆异黄酮还原酶（GmIFR）家族含有17个成员，蛋白序列介于191（GmIFR07）和376（GmIFR11）个氨基酸之间，序列相似性为19.59%（GmIFR07和GmIFR11）～98.43%（GmIFR12和GmIFR17），结构分析表明这些基因内含子数目差异较大，2～6个不等，不均匀的分布在大豆的1、4、6、9、11、12和16号染色体上。
Abstract:
Isoflavones are a class of secondary metabolites formed in soybean growth, which are the main functional active components of soybean.Isoflavone reductase is one of the key enzymes catalyzed isoflavone decomposition and plays an important role in the regulation of isoflavone content and composition. In this research, we investigated the isoflavone reductase (IFR) gene family based on the results of soybean whole genome sequencing and analyzed the conservation of the sequence, sequence characteristics, structural features, genetic diversity as well as their phylogenetic relationships. The result showed that there were 17 gene members at least existed in the soybean genome with protein sequence length varying from 191 to 376. Sequence identity analysis showed that the lowest value of sequence similarity was observed between GmIFR07 and GmIFR11(19.59%) and the highest value was observed between GmIFR12 and GmIFR17(98.43%).Gene structure analysis showed the number of introns among these genes was ranging from 2 to 6- Moreover, these IFR gene family members distribute in chromosome 1, 4, 6, 9, 11, 12 and 16 in soybean unequally.

参考文献/References:

［1］Xu B, Chang S K C. Characterization of phenolic substances and antioxidant properties of food soybeans grown in North DakotaMinnesota Region［J］. Journal of Agricultural and Food Chemistry, 2008, 56(19): 9102-9113．

［2］Ward H A, Kuhnle G G, Mulligan A A, et a1. Breast, colorectal, and prostate cancer risk in the European Prospective Investigation into cancer and nutrition-norfolk in relation to phytoestregen intake derived from an improved database［J］.The American Journal of Clinical Nutrition, 2010, 91(2): 440-448.
［3］Li S H, Liu X X, Bai Y Y, et a1.Effect of oral isoflavone supplementation on vascular endothelial function in postmenopausal women: A meta analysis of randomized placebocontrolled trials［J］. The American Journal of Clinical Nutrition, 2010, 91(2): 480-486.
［4］Cheng H, Yu O, Yu D Y. Polymorphisms of IFS1 and IFS2 gene are associated with isoflavone concentrations in soybean seeds［J］.Plant Science, 2008, 175(4): 505-512.
［5］Lozovaya V V, Lygin A V, Zernova O V, et al. Modification of phenolic metabolism in soybean hairy roots through down regulation of chalcone synthase or isoflavone synthase［J］. Planta, 2007, 225(3): 665-679.
［6］Subramanian S, Hu X, Lu G H, et al. The promoters of two isoflavone synthase genes respond differentially to nodulation and defense signals in transgenic soybean roots［J］. Plant Molecular Biology, 2004, 54(5): 623-639.
［7］Chiari L, Piovesan N D, Naoe L K, et al. Genetic parameters relating isoflavone and protein content in soybean seeds［J］.Euphytica, 2004, 138(1): 55-60.
［8］Dixon R A, Steele C L. Flavonoids and isoflavonoids-a gold mine for metabolic engineering［J］. Trends in Plant Science, 1999, 4(10): 394-400.
［9］乞永燕, Sofia M, Sjef B, 等. 大豆中Glycinol的分离与鉴定［J］. 色谱, 2005, 23(4): 353-357. (Qi Y Y, Sofia M, Sjef B, et a1. Isolation and identification of Glycinol from Glycine max［L］Merri［J］. Chinese Journal o f Chromatography, 2005, 23(4): 353-357.)
［10］李炜, 来永才, 毕远林, 等. 黑龙江省野生大豆高异黄酮新种质创新利用研究Ⅱ异黄酮含量与大豆品质相关性的分析［J］. 大豆科学, 2007, 26(3): 319-321. (Li W, Lai Y C, Bi Y L, et al.Innovation and utilization of new high isoflavone resource of wild soybean in Heilongjiang Ⅱ Analysis of relationship between isoflavone content and quality of soybean［J］. Soybean Science, 2007, 26(3): 319-321.)
［11］Cheng H, Li L L, Xu F, et al.Expression patterns of an isoflavone reductase-like gene and its possible roles in secondary metabolism in Ginkgo biloba［J］. Plant Cell Reports, 2013, 32(5): 637-650.
［12］谢晓娜, 张小秋, 邵敏, 等.甘蔗类异黄酮还原酶(IRL)基因的克隆与表达分析［J］. 生物技术通报, 2014, 11: 130-137.(Xie X N, Zhang X Q, Shao M, et al. Cloning and expression analysis of sugarcane isoflavone reductase-like(IRL) gene［J］. Biotechnology Bulletin, 2014, 11: 130-137.)
［13］Babaoglu M, Davey M R, Power J B, et al.Transformed roots of Lupinus mutabilis: Induction, culture and isoflavone biosynthesis［J］. Plant Cell, Tissue and Organ Culture, 2004, 78(1): 29-36.［14］Schmutz J, Cannon S B, Schlueter J, et a1. Genome sequence of the palaeopolyploid soybean［J］.Nature, 2010, 463(7278): 178-183.
［15］王荣青, 万红建, 李志邈, 等. 番茄Hsp70基因鉴定及系统发育关系分析［J］. 核农学报, 2014, 28(3): 378-385. (Wang R Q, Wan H J, Li Z M, et al. Identification and phylogenetic relationships of Hsp70 genes in tomato［J］. Journal of Nuclear Agricultural Sciences, 2014, 28(3): 378- 385.)
［16］张古文, 刘莉莉, 王显瑞, 等. 谷子HSP70基因家族的全基因组鉴定及生物信息学分析［J］. 浙江农业学报, 2015, 27(7): 1127-1133. (Zhang G W, Liu L L, Wang X R, et al. Genome-wide identification and bioinformatics analysis of HSP70 genes in foxtail millet［J］. Acta Agriculturae Zhejiangensis, 2015, 27(7): 1127- 1133.)
［17］Tsukamoto C, Shimada S, Igita K, et al. Factors affecting isoflavone content in soybean seeds: Changes in isoflavones, saponins and composition of fatty acids at different temperarure during seed development［J］. Journal of Agricultural & Food Chemistry, 1995, 43(5): 1184-1192.
［18］罗燕, 郭万柱, 刘艳丽, 等.猪细小病毒SC-1株VP2基因的克隆及生物信息学分析［J］.动物医学进展, 2005, 26(8): 87-91. (Luo Y, Guo W Z, Liu Y L, et al. The cloning and bioinformation analysis of VP2 gene of porcine parvovirus strain SC-1［J］Progress in Veterinary Medicine, 2005, 26(8): 87-91.)
［19］赵海霞, 李成磊, 白悦辰, 等. 苦荞类异黄酮还原酶基因(FtIRL)的克隆及序列分析［J］.食品科学, 2012, 33(11): 210-214.(Zhao H X, Li C L, Bai Y C, et al. Cloning and sequence analysis of the isoflavone reductase-like (FtIRL) gene from .Fagopyrum tatarium［J］. Food Science, 2012, 33(11): 210-214.)
［20］陈伟, 林叶春, 高维常, 等. 烟草类异黄酮还原酶基因家族的分子克隆［J］. 中国农业大学学报, 2014, 19(6): 23-34. (Chen W, Lin Y C, Gao W C, et al. Molecular cloning of isoflavone reductase-like gene family from tobacco［J］. Journal of China Agricultural University, 2014, 19(6): 23-34.)
［21］李乐, 许红亮, 杨兴露, 等.大豆-LEA-基因家族全基因组鉴定、分类和表达［J］. 中国农业科学, 2011, 44(19): 3945-3954.(Li L, Xu H L, Yang X L, et al. Genome-wide identification, classification and expression analysis of LEA gene family in soybean［J］. Scientia Agricultura Sinica, 2011, 44(19): 3945-3954.)
［22］Li Y H, Zhou G, Ma J, et al. De novo assembly of soybean wild relatives for pan-genome analysis of diversity and agronomic traits［J］.Nature Biotechnology, 2014, 32(10): 1045-1052.
［23］Zhu Q L, Guo T Y, Sui S Z, et al.Molecular cloning and characterization of a novel isoflavone reductase-like gene (FcIRL) from high flavonoids-producing callus of Fagopyrum cymosum［J］.Acta Pharmaceutica Sinica, 2009, 44(7): 809-819.
［24］Cheng H, Li L, Xu F, et al. Expression patterns of an isoflavone reductase-like gene and its possible roles in secondary metabolism in Ginkgo biloba［ J］. Plant Cell Reports, 2013, 32(5): 637-650.
［25］Nasiri J, Naghavi M R, Alizadeh H, et al. Seasonal-based temporal changes fluctuate expression patterns of TXS, DBAT, BAPT and DBTNBT genes alongside production of associated taxanes in .Taxus baccata［J］. Plant Cell Reports, 2016, 35(5): 1103-1119.
［26］Shoji T, Winz R, Iwase T, et al. Expression patterns of two tobacco isoflavone reductase-like genes and their possible roles in secondary metabolism in tobacco［J］. Plant Molecular Biology, 2002, 50(3): 427-440.

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备注/Memo

基金项目：浙江省重大科技专项（2012C12903-4-1）；浙江省植物有害生物防控重点实验室—省部共建国家重点实验室培育基地项目（2010DS700124-KF1611）；浙江省公益技术应用研究计划项目（2015C32050）。

第一作者简介：张古文(1981-)，男，博士，主要从事菜用大豆遗传育种和分子标记开发研究。E-mail: zhangguwen@126.com。通讯作者：龚亚明（1969-），男，研究员，主要从事菜用豆类育种和推广工作。 E-mail: gongym07@126.com

更新日期/Last Update: 2016-12-08