[1]廖春梅,陈丽玉,杨涔,等.大豆NIN和NLP基因生物信息学分析及敲除载体构建[J].大豆科学,2023,42(01):1-11.[doi:10.11861/j.issn.1000-9841.2023.01.0001]
点击复制

大豆NIN和NLP基因生物信息学分析及敲除载体构建

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第42卷 期数: 2023年01期 页码: 1-11 栏目: 出版日期: 2023-01-20
作者:
廖春梅; 陈丽玉; 杨涔; 刘宝辉; 孔凡江
广州大学 生命科学学院/分子遗传与进化创新研究中心， 广东 广州 510030
关键词:
大豆; 根瘤; NLP基因; 生物信息学分析; CRISPR/Cas 9基因编辑技术; 基因敲除载体; GmNLP4a/b; GmNLP5a/b
DOI:
10.11861/j.issn.1000-9841.2023.01.0001
摘要:
为促进大豆NLP家族基因突变大豆材料的获得及大豆NIN和NLP基因功能的研究，本研究对大豆NIN和NLP基因进行生物信息学分析，通过CRISPR/Cas 9基因编辑技术构建基因敲除载体，并且通过大豆毛根转化实验验证靶点的有效性。结果表明：大豆基因组中一共存在4个NIN基因和10个NLP基因家族成员，这些基因都具有RWP-RK和PB1两个保守结构域。亚细胞定位预测表明所有成员都定位在细胞核中，此外GmNIN1b和GmNIN2a还定位于叶绿体。GmNIN1a/b、GmNIN2a/b、GmNLP2a/b及GmNLP3b在根瘤中的表达量相对较高，推测这些基因可能对结瘤过程具有重要的调控功能。成功构建了NLP4和NLP5两个敲除载体，得到可敲除GmNLP4a/b的3个有效靶点和敲除GmNLP5a/b的2个有效靶点。本研究获得了大豆NIN和NLP基因家族的生物信息学依据和创制GmNLP4a/b和GmNLP5a/b基因突变体的技术依据。

相似文献/References:

[1]刘章雄,李卫东,孙石,等.1983～2010年北京大豆育成品种的亲本地理来源及其遗传贡献[J].大豆科学,2013,32(01):1.[doi:10.3969/j.issn.1000-9841.2013.01.002]
　LIU Zhang-xiong,LI Wei-dong,SUN Shi,et al.Geographical Sources of Germplasm and Their Nuclear Contribution to Soybean Cultivars Released during 1983 to 2010 in Beijing[J].Soybean Science,2013,32(01):1.[doi:10.3969/j.issn.1000-9841.2013.01.002]
[2]李彩云,余永亮,杨红旗,等.大豆脂质转运蛋白基因GmLTP3的特征分析[J].大豆科学,2013,32(01):8.[doi:10.3969/j.issn.1000-9841.2013.01.003]
　LI Cai-yun,YU Yong-liang,YANG Hong-qi,et al.Characteristics of a Lipid-transfer Protein Gene GmLTP3 in Glycine max[J].Soybean Science,2013,32(01):8.[doi:10.3969/j.issn.1000-9841.2013.01.003]
[3]王明霞,崔晓霞,薛晨晨,等.大豆耐盐基因GmHAL3a的克隆及RNAi载体的构建[J].大豆科学,2013,32(01):12.[doi:10.3969/j.issn.1000-9841.2013.01.004]
　WANG Ming-xia,CUI Xiao-xia,XUE Chen-chen,et al.Cloning of Halotolerance 3 Gene and Construction of Its RNAi Vector in Soybean (Glycine max)[J].Soybean Science,2013,32(01):12.[doi:10.3969/j.issn.1000-9841.2013.01.004]
[4]张春宝,李玉秋,彭宝,等.线粒体ISSR与SCAR标记鉴定大豆细胞质雄性不育系与保持系[J].大豆科学,2013,32(01):19.[doi:10.3969/j.issn.1000-9841.2013.01.005]
　ZHANG Chun-bao,LI Yu-qiu,PENG Bao,et al.Identification of Soybean Cytoplasmic Male Sterile Line and Maintainer Line with Mitochondrial ISSR and SCAR Markers[J].Soybean Science,2013,32(01):19.[doi:10.3969/j.issn.1000-9841.2013.01.005]
[5]卢清瑶,赵琳,李冬梅,等.RAV基因对拟南芥和大豆不定芽再生的影响[J].大豆科学,2013,32(01):23.[doi:10.3969/j.issn.1000-9841.2013.01.006]
　LU Qing-yao,ZHAO Lin,LI Dong-mei,et al.Effects of RAV gene on Shoot Regeneration of Arabidopsis and Soybean[J].Soybean Science,2013,32(01):23.[doi:10.3969/j.issn.1000-9841.2013.01.006]
[6]杜景红,刘丽君.大豆fad3c基因沉默载体的构建[J].大豆科学,2013,32(01):28.[doi:10.3969/j.issn.1000-9841.2013.01.007]
　DU Jing-hong,LIU Li-jun.Construction of fad3c Gene Silencing Vector in Soybean[J].Soybean Science,2013,32(01):28.[doi:10.3969/j.issn.1000-9841.2013.01.007]
[7]张力伟,樊颖伦,牛腾飞,等.大豆“冀黄13”突变体筛选及突变体库的建立[J].大豆科学,2013,32(01):33.[doi:10.3969/j.issn.1000-9841.2013.01.008]
　ZHANG Li-wei,FAN Ying-lun,NIU Teng-fei?,et al.Screening of Mutants and Construction of Mutant Population for Soybean Cultivar "Jihuang13”[J].Soybean Science,2013,32(01):33.[doi:10.3969/j.issn.1000-9841.2013.01.008]
[8]盖江南,张彬彬,吴瑶,等.大豆不定胚悬浮培养基因型筛选及基因枪遗传转化的研究[J].大豆科学,2013,32(01):38.[doi:10.3969/j.issn.1000-9841.2013.01.009]
　GAI Jiang-nan,ZHANG Bin-bin,WU Yao,et al.Screening of Soybean Genotypes Suitable for Suspension Culture with Adventitious Embryos and Genetic Transformation by Particle Bombardment[J].Soybean Science,2013,32(01):38.[doi:10.3969/j.issn.1000-9841.2013.01.009]
[9]王鹏飞,刘丽君,唐晓飞,等.适于体细胞胚发生的大豆基因型筛选[J].大豆科学,2013,32(01):43.[doi:10.3969/j.issn.1000-9841.2013.01.010]
　WANG Peng-fei,LIU Li-jun,TANG Xiao-fei,et al.Screening of Soybean Genotypes Suitable for Somatic Embryogenesis[J].Soybean Science,2013,32(01):43.[doi:10.3969/j.issn.1000-9841.2013.01.010]
[10]刘德兴,年海,杨存义,等.耐酸铝大豆品种资源的筛选与鉴定[J].大豆科学,2013,32(01):46.[doi:10.3969/j.issn.1000-9841.2013.01.011]
　LIU De-xing,NIAN Hai,YANG Cun-yi,et al.Screening and Identifying Soybean Germplasm Tolerant to Acid Aluminum[J].Soybean Science,2013,32(01):46.[doi:10.3969/j.issn.1000-9841.2013.01.011]
[11]张立军,孙旭刚,王昌陵,等.盆栽条件下水肥调控对大豆生长和产量的影响[J].大豆科学,2014,33(03):398.[doi:10.11861/j.issn.1000-9841.2014.03.0398]
　ZHANG Li-jun,SUN Xu-gang,WANG Chang-ling,et al.Regulation Effect of Water and Fertilizer on Growth and Yield in Soybean under Pot Experiment Condition[J].Soybean Science,2014,33(01):398.[doi:10.11861/j.issn.1000-9841.2014.03.0398]
[12]闫春娟,王文斌,孙旭刚,等.水肥互作对大豆生理特性、根瘤固氮及产量的影响[J].大豆科学,2011,30(02):229.[doi:10.11861/j.issn.1000-9841.2011.02.0229]
　YAN Chun-juan,WANG Wen-bin,SUN Xu-gang,et al.Effect of Water-Fertilizer Interaction on Physiological Characteristics，Nitrogen Fixation and Yield of Soybean[J].Soybean Science,2011,30(01):229.[doi:10.11861/j.issn.1000-9841.2011.02.0229]
[13]乔云发,韩晓增.长期定量施肥对大豆根系形态和根瘤性状的影响[J].大豆科学,2011,30(01):119.[doi:10.11861/j.issn.1000-9841.2011.01.0119]
　QIAO Yun-fa,HAN Xiao-zeng.Effects of Long-term Fertilization on Root Phenotype and Nodulation of Soybean[J].Soybean Science,2011,30(01):119.[doi:10.11861/j.issn.1000-9841.2011.01.0119]
[14]于晓波,向达兵,杨文钰,等.结瘤性状不同基因型大豆对接种花生根瘤菌Spr2-9的响应[J].大豆科学,2010,29(01):41.[doi:10.11861/j.issn.1000-9841.2010.01.0041]
　YU Xiao-bo,XIANG Da-bing,YANG Wen-yu,et al.Response of Soybean Genotypes with Different Nodulation Traits to Peanut Rhizobia Spr2-9 Inoculation[J].Soybean Science,2010,29(01):41.[doi:10.11861/j.issn.1000-9841.2010.01.0041]
[15]严君,韩晓增,王守宇,等.不同形态氮对大豆根瘤生长及固氮的影响[J].大豆科学,2009,28(04):674.[doi:10.11861/j.issn.1000-9841.2009.04.0674]
　YAN Jun,HAN Xiao-zeng,WANG Shou-yu,et al.Effect of Different Forms Nitrogen on Nodule Growth and Nitrogen Fixation in Soybean (Glycine max L.)[J].Soybean Science,2009,28(01):674.[doi:10.11861/j.issn.1000-9841.2009.04.0674]
[16]蒋跃林 　张庆国 　张仕定 　岳 　伟 　陈庭甫 　樊丽莉.大气 CO 2 浓度升高对大豆根瘤量及其固氮活性的影响＊[J].大豆科学,2006,25(01):53.[doi:10.11861/j.issn.1000-9841.2006.01.0053]
　Jiang Yuelin Zhang Qingguo Zhang Shiding Yue Wei Chen Tinfu Fan Lili.EFFECTOF ELEVATED ATMOSPHERIC CO 2 CONCENTRATION ONROOT NODULES AND NITROGENASE ACTIVITY IN SOYBEAN[J].Soybean Science,2006,25(01):53.[doi:10.11861/j.issn.1000-9841.2006.01.0053]
[17]於丙军　刘友良.大豆耐盐性研究进展[J].大豆科学,2000,19(02):154.[doi:10.11861/j.issn.1000-9841.2000.02.0154]
　Yu BingjunLiu Yo uliang.RECENT PROGRESS IN STUDY ON SALT TOLERANCE IN SOYBEAN[J].Soybean Science,2000,19(01):154.[doi:10.11861/j.issn.1000-9841.2000.02.0154]
[18]张明才　何钟佩　田晓莉　王保民　段留生　翟志席　李召虎.植物生长调节剂BR 和SHK -6 对大豆生物产量和根瘤固氮活性的激素调控研究1[J].大豆科学,2004,23(02):96.[doi:10.11861/j.issn.1000-9841.2004.02.0096]
　Zhang Ming cai He Zhongpei Tian Xiaoli Wang Baomin Duan liusheng Zhai zhixi Li Zhaohu.HORMONAL REGULATION OF PLANT GROWTH REGULATOR BR ANDSHK-6 ON SOYBEAN BIOMASS AND NITROGENASE ACTIVITY[J].Soybean Science,2004,23(01):96.[doi:10.11861/j.issn.1000-9841.2004.02.0096]
[19]甘银波　陈静　Ineke Stulen.大豆不同生长阶段施用氮肥对生长、结瘤及产量的影响[J].大豆科学,1997,16(02):125.[doi:10.11861/j.issn.1000-9841.1997.02.0125]
　Gan Yinbo Chen Jing Ineke Stulen.EFFECTS OF N APPLTCATION AT DTFFERENT GROWTH STAGES ON GROWTHNODULATIONAND YTELD OF SOYBEANS[J].Soybean Science,1997,16(01):125.[doi:10.11861/j.issn.1000-9841.1997.02.0125]
[20]姬月梅,罗瑞萍,赵志刚,等.不同施氮量及施氮方式对大豆根瘤生长及产量的影响[J].大豆科学,2017,36(06):887.[doi:10.11861/j.issn.1000-9841.2017.06.0887]
　JI Yue-mei,LUO Rui-ping,ZHAO Zhi-gang,et al.Effects of Different Nitrogen Application and Methods on Nodule Growth and Yield of Soybean (Glycine max L.)[J].Soybean Science,2017,36(01):887.[doi:10.11861/j.issn.1000-9841.2017.06.0887]

备注/Memo

广东省基础与应用基础研究重大项目（2019B030302006）。

更新日期/Last Update: 2023-01-20