[1]秦玉涛,王瑞,吕金东,等.大豆炸荚基因GmAGL8的转基因鉴定与功能初探[J].大豆科学,2017,36(03):360-364.[doi:10.11861/j.issn.1000-9841.2017.03.0360]
　QIN Yu-tao,WANG Rui,LYU Jin-dong,et al.Transgenic Identification of Pod-shattering Gene GmAGL8 in Soybean[J].Soybean Science,2017,36(03):360-364.[doi:10.11861/j.issn.1000-9841.2017.03.0360]
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大豆炸荚基因GmAGL8的转基因鉴定与功能初探

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第36卷 期数: 2017年03期 页码: 360-364 栏目: 出版日期: 2017-05-20
Title:
Transgenic Identification of Pod-shattering Gene GmAGL8 in Soybean
作者:
秦玉涛1; 王瑞2; 吕金东1; 田源2; 黄天珑2; 谢皓1; 郭蓓2; 3
（1.北京农学院 植物科学技术学院，北京 102206； 2.北京农学院 生物科学与工程学院，北京 102206； 3.中华人民共和国农业部华北都市农业重点实验室，北京 102206）
Author(s):
QIN Yu-tao1;  WANG Rui2;  LYU Jin-dong1;  TIAN Yuan2;  HUANG Tian-long2;  XIE Hao1;  GUO Bei2; 3
(1.College of Plant Science and technology, Beijing University of Agriculture, Beijing 102206, China; 2.College of Biological Science and Engineering, Beijing University of Agriculture, Beijing 102206, China; 3.Key Laboratory of Urban Agriculture (North China), Ministry of Agriculture, P R, Beijing 102206, China)
关键词:
大豆; 炸荚; GmAGL8; 转基因
Keywords:
Soybean;  Pod-shattering;  GmAGL8;  Transgene
DOI:
10.11861/j.issn.1000-9841.2017.03.0360
文献标志码:
A
摘要:
炸荚是大豆的一种自然特征属性，是影响大豆产量的重要因素之一。本研究以前期获得的转大豆炸荚相关基因GmAGL8的T1代植株为材料，继续繁育获得T2和T3代；采用PCR和RT-qPCR检测方法对转基因植株进行基因遗传稳定性和表达情况分析；并以野生型中黄10号为对照，对大豆炸荚性状进行了鉴定分析。结果表明：转基因植株阳性率T1代为87.5％，T2和T3代均达到100%，说明GmAGL8基因已基本能够在转基因后代中稳定遗传。RT-qPCR检测结果显示，转基因植株中GmAGL8基因的相对表达量都明显高于非转基因植株，且各转基因植株之间表达量具有差异性。对T1、T2和T3代植株炸荚率进行了统计，不同世代转基因大豆的平均炸荚率为9.09％，而非转基因大豆炸荚率为83.3％，转基因与非转基因大豆之间炸荚率存在显著差异。综上所述，GmAGL8基因已基本实现在大豆转基因后代中稳定遗传并正常表达，表型鉴定结果初步证明了GmAGL8基因与大豆炸荚性状相关。
Abstract:
Pod shattering is a natural characteristic of soybean, which is one of the critical factors influencing production. In our previous study, we had transformed the gene GmAGL8(which putative related to soybean pod-shattering) into Glycine max and obtained the T1generation.In this study, the T 2 and T3 generation were multiplied by the T 1 generation carrying the GmAGL8-gene.The genetic stability and expression of transgenic plants were analyzed by using PCR and RT-qPCR methods. As a control group, wild-type Zhonghuang 10 was used to analyze and identify the traits of soybean pods.The results showed the positive rate of the GmAGL8 gene in T1, T2 and T 3 generation were 87.5%, 100% and 100% respectively.It demonstrated that the GmAGL8 gene can be inherited stably.The results of RT-qPCR analyzing showed the relative expression amount of GmAGL8-gene in transgenic plants was significantly higher than that in non-transgenic plants, and the amounts were various among the transgenic plants. The statistical results for pod-shattering among all plants indicated that the rates of pod-shattering were different among transgenic plants and non-transgenic plants, the average rate of pod shattering among transgenic plants of all generations was 9.09%, the rate of pod shattering among non-transgenic plants was 83.3%.In conclusion, GmAGL8 gene can be inherited stably and expressed normally among transgenic soybean.Furthermore, the phenotypic identification results proved the GmAGL8 gene is related to pod-shattering.

参考文献/References:

［1］董钻.大豆产量生理［M］. 北京: 中国农业出版社, 2000: 17-18. (Dong Z.Soybean yield physiology［M］. Beijing: Agriculture Press, 2000, 17-18)

［2］王连铮, 郭庆元. 现代中国大豆［M］. 北京: 金盾出版社, 2007: 199-200. (Wang L Z, Guo Q Y. Modern Chinese soybean［M］. Beijing: Jindun Press, 2007: 199-200.)
［3］Philbrook B D, Oplinger E S. Soybean field losses as influenced by harvest delays［J］. Agronomy Journal, 1989, 81(2): 251-258.
［4］张跃进, 马赛斐, 高启云, 等. 黄淮流域主栽大豆品种炸荚性研究［J］. 河南农业科学, 2006(6): 56-59. (Zhang J Y, Ma S F, Gao Q Y, et al.Study on the podshattering of main soybean varieties of Huanghuai area［J］.Journal of Henan Agricultural Sciences, 2006(6): 56-59.)
［5］Theissen G, Becker A, Di Rosa A, et al.Ａ short history of MADS-box genes in［J］. Plant Molecular Biology, 2000, 42(1): 115-149.
［6］Becker A, Winter K U, Meyer B, et al.MADS-box gene diversity in seed plants 300 million years ago［J］. Molecular Biology and Evolution, 2000, 17(10): 1425-1434.
［7］黄方, 迟英俊, 喻得跃. 植物MADS-box基因研究进展［J］. 南京农业大学学报, 2012, 35(5): 9-18. (Huang F, Chi Y J, Yu D Y. Research advances of MADS-box genes in plants［J］. Journal of Nanjing Agricultural University, 2012, 35(5): 9-18.)
［8］Fang H, Guang L X, Ying J H, et al.A soybean MADS-box protein modulates floral organ numbers, petal identity and sterility［J］. BMC Plant Biology, 2014,14: 89
［9］褚婷婷, 谢华, 徐勇, 等. 植物MADS-box基因FRUITFULL(FUL)研究进展［J］. 中国生物工程杂志, 2010, 30(9):98-104. (Zhe T T, Xie H, Xu Y, et al. Research advances of plant MADS-box gene FRUITFULL(FUL)［J］. China Biotechnology, 2010, 30(9): 98-104.)
［10］徐勇, 张林, 马荣才. 桃(Prunus persica)中2个MADSbox基因功能的初步鉴定和遗传作图［J］. 科学通报, 2008, 53(5) : 537-543.(Xu Y, Zhang L, Ma R C.Function preliminary evaluation and genetic mapping of two MADSbox gene in peach(Prunus persica)［J］. Chinese Science Bulletin, 2008, 53(5) : 537-543.)
［11］Hempel F D, Weigel D, Mandel M A, et al. Floral determination and expression of floral regulatory genes in Arabidopsis［J］. Development, 1997, 124: 3845-3853.
［12］Ferrandiz C, Gu Q, Martienssen R, et al.Redundant regulation of meristem identity and plant architecture by FRUITFULL, APETALA1and CAULIFLOWER［J］. Development, 2000, 127: 725-734.
［13］李冉阳, 秦玉涛, 张倩, 等. 大豆GmAGL8基因的克隆及生物信息学分析［J］. 安徽农业科学, 2015, 43(18): 45-48.(Li R Y, Qin Y T, Zhang Q, et al. Cloning and bioinfirmatice analysis of gene GmAGL8 in soybean［J］. Journal of Anhui Agricultural Sciences, 2015, 43(18): 45-48.)
［14］Lerat S, England L S, Vincent M L, et al. Real-time polymerase chain reaction quantification of the transgenes for roundup ready corn and roundup ready soybean in soil samples［J］. Journal of Agricultural and Food Chemistry, 2005, 53(5): 1337-1342.
［15］王海慧, 田源, 白国辉, 等. 转基因植物遗传稳定性影响因素的研究现状［J］. 贵州农业科学, 2013(9): 5-8.(Wang H H, Tian Y, Bai G H, et al.Adances in infulencing factors of genetic stability of transgenic plants［J］. Guizhou Agricultural Sciences, 2013(9): 5-8.)
［16］彭玉华, 袁建中, 杨国保. 黄淮流域大豆炸荚性初步分析［J］. 大豆科学, 1991, 10(4): 285-230. (Peng Y H, Yuan Z J, Yang B G. Primary analysis of soybean shattering in Chineses Yellow River soybean adaptaion region［J］. Soybean Science, 1991, 10(4): 285-230.)
［17］马赛斐, 南翠梅, 张凤彩, 等. 黄淮流域主栽大豆品种炸荚性研究初报［J］. 大豆通报, 2006 (2): 21-23. (Ma S F, Nan C M, Zhang F C, et al. Preliminary investigation on pod-shattering of main soybean cultivars in Huanghuai area［J］. Soybean Bulletin, 2006(2): 21-23.)
［18］Kang S T, Kwak M, Kim H K, et al. Population-specific QTL sand their different epistatic interactions for pod dehiscencein soybean［Glycine max(L) Merr］［J］. Euphytica, 2009,166(1): 15-24.

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备注/Memo

基金项目：国家高技术研究发展计划“863计划”（2012AA101106）。

第一作者简介：秦玉涛（1990-），男，硕士，主要从事大豆分子遗传育种研究。E-mail: 372766399@qq.com。
通讯作者：郭蓓（1962-），女，博士，教授，主要从事植物分子遗传育种研究。E-mail: guobeibac@sohu.com。

更新日期/Last Update: 2017-06-24