接种红色荧光蛋白标记根瘤菌对苜蓿幼苗生长及结瘤能力的影响
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摘要
将含有红色荧光蛋白(RFP)基因质粒的荧光标记根瘤菌S.LZgn5-rfp2,S.12531-rfp4、S.LH343-rfp3接种于甘农5号和WL343HQ紫花苜蓿,分别测定2个苜蓿品种幼苗的生物量、生长指标、固氮酶活性等指标。结果表明:RFP标记根瘤菌与出发菌相比对苜蓿幼苗生长无显著影响,与不接菌的苜蓿幼苗相比有明显的促生效果,能稳定的在苜蓿幼苗体内表达,并具有与出发菌无差异的促生能力,2个苜蓿品种间也无特异性。
Red fluorescent protein marked rhizobias(S.LZgn5-rfp3、S.LZgn5-rfp4、S.LZgn5-rfp2,S.12531-rfp3、S.12531-rfp4、S.12531-rfp2、S.LH343-rfp3、S.LH343-rfp4、S.LH343-rfp2) that contained the plasmid with RFP(red fluorescent protein) gene were inoculated to Gannong No.5 alfalfa and alfalfa WL343 HQ.Biomass,nodule number,nitrogenase activity and the rate of effective nodules were measured.The results showed that the RFP fluorescence-marked rhizobias could promote plant growth and biomass accumulation of inoculated alfalfa.Compared with their corresponding original rhizobias,the RFP fluorescence-marked rhizobias did no injury the alfalfa seedling,could be expressed consistently in alfalfa plant,had no difference from the starting bacteria,had the similar growth promotion ability with original rhizobias.It could be stably expressed in alfalfa seedlings and had no difference from the starting bacteria.Considering ability,there is no specificity between the two alfalfa varieties.
Red fluorescent protein marked rhizobias(S.LZgn5-rfp3、S.LZgn5-rfp4、S.LZgn5-rfp2,S.12531-rfp3、S.12531-rfp4、S.12531-rfp2、S.LH343-rfp3、S.LH343-rfp4、S.LH343-rfp2) that contained the plasmid with RFP(red fluorescent protein) gene were inoculated to Gannong No.5 alfalfa and alfalfa WL343 HQ.Biomass,nodule number,nitrogenase activity and the rate of effective nodules were measured.The results showed that the RFP fluorescence-marked rhizobias could promote plant growth and biomass accumulation of inoculated alfalfa.Compared with their corresponding original rhizobias,the RFP fluorescence-marked rhizobias did no injury the alfalfa seedling,could be expressed consistently in alfalfa plant,had no difference from the starting bacteria,had the similar growth promotion ability with original rhizobias.It could be stably expressed in alfalfa seedlings and had no difference from the starting bacteria.Considering ability,there is no specificity between the two alfalfa varieties.
引文
[1] Matz M V,Fradkov,Labas Y A,et al.Fluorescent proteins from nonbioluminescentAnthozoa species[J].Nat Biotechnol,1999,17(10):969-973.
[2] 王飞,杨海涛,王泽方.红色荧光蛋白的研究进展[J].生物技术通报,2017,33(9):32-47.
[3] 杨晓玫.红、黄、绿荧光蛋白标记根瘤菌的构建[D].兰州:甘肃农业大学硕士论文,2017.
[4] 杨晓玫,师尚礼.红、黄、绿三种颜色荧光质粒导入大肠杆菌中的稳定性表达[J].甘肃农业大学学报,2018,53(3):193-198.
[5] 杨晓玫,周彤,阿芸,等.苜蓿根瘤菌GFP荧光标记株的构建及对结瘤固氮的影响[J].草原与草坪,2018,38(1):44-56.
[6] Tan G Y.Genetic variation for acetylene reduction rate and other character in alfalfa[J].Crop Science,1981,21:485-488.
[7] 张虎天,郭丽琢,柴强,等.接种根瘤菌对豌豆/玉米体系根际细菌数量及氮营养的影响[J].甘肃农业大学学报,2011,46(1):30-33.
[8] 陈力玉,张淑卿,李剑锋,等.接种荧光标记根瘤菌对苜蓿幼苗生长的影响[J].草原与草坪,2013,33(6):1-8.
[9] 袁全,杨梦华,郑会明,等.中慢生型华癸根瘤菌Mesorhizobiumhuakuii AS9 自体诱导物合成酶基因的筛选及其功能分析[J].土壤,2009,41(3):459-463.
[10] 殷爱华,韩素芬.豆科树种凝集素和根瘤菌胞外多糖结合反应与结瘤的关系[J].南京林业大学学报(自然科学版),2005,29(5):88-90.
[11] Hoagland D,Arnon D I.The water culture method for growing plants without soil[J].California Agricultural Experiment Station Bulletin,1938(1):1-39.
[12] 高永革,李黎,刘祥等.黄河滩区紫花苜蓿生产性能比较研究[J].草业科学,2008,25(7):59-64.
[13] Vertucci C W,Roosee,Crane J.The oretical basis of protocols foe seed storage optimummoisture contents for pea seeds stored at different temperature[J].Annalsof Botany,1994,74:531-540.
[14] 李剑峰,师尚礼,张淑卿.环境酸度对紫花苜蓿早期生长和生理的影响[J].草业学报,2010,19(2):47-54.
[15] Hara S,Hashidoko Y,Desyatkin R V,et al.High rate of N2 fixation by east Siberian cryophilic soil bacteria as determined by measuring acetylene reduction in nitrogen poor medium solid ified with gellan gum[J].Appliedand Environmental Microbiology,2009,75:2811-2819.
[16] 谭志远,彭桂香,徐培智,等.普通野生稻(Oryzarufipogen)内生固氮菌多样性及高固氮酶活性[J] .科学通报,2009,54:1885-1893.
[17] 陈力玉.基于三亲本杂交的荧光标记根瘤菌的构建及其稳定性检测研究[D].兰州:甘肃农业大学硕士论文,2013.
[18] 邹琦.植物生理生化实验指导[M].北京:中国农业出版社,1995.
[19] Muhammad Z,AsifT,Ata C Z,et al.Weed-crop competition effects on growth and yield of sugarcane planted using two methods[J].Pakistan Journal of Botany,2010,42(2):815-823.
[20] 霍尔特(著).刘复今(译).伯杰鉴定手册[M](第八版).济南:山东大学出版社,1988:93-94.
[21] 张淑卿.根瘤菌在苜蓿植株体内的运移及影响因素[D].兰州:甘肃农业大学,2012.
[22] 陈翠翠,马元武,冯永君,等.MADS-box 家族蛋白在植物开花、结实及根瘤形成中的多功能调节作用[J].华北农学报,2008,23(S2):80-83.
[23] DudejaSS.PersistenceofDradyrhizobiumSP.(caianus)inasandyloam[J].SoilBoilBiochem,1989,21(5):709-713.
[24] 凌瑶,张小平,周俊初,等.用荧光酶基因(cfp)标记法研究菜豆根瘤菌的竞争性和有效性[J].中国土壤与肥料,2008(1):73-77.
[25] 韩颖,赵寿经,杨瑜,等,玉米陈化关键酶基因Zmlox-2的RNA干扰载体的构建[J].华南理工大学学报(自然科学版),2016,44(3):136-140.
[26] 董伟欣,张月辰,张迎迎.DsRED红色荧光蛋白植物表达载体的构建和表达[J].河北农业大学学报,2010,5(3):33.
[2] 王飞,杨海涛,王泽方.红色荧光蛋白的研究进展[J].生物技术通报,2017,33(9):32-47.
[3] 杨晓玫.红、黄、绿荧光蛋白标记根瘤菌的构建[D].兰州:甘肃农业大学硕士论文,2017.
[4] 杨晓玫,师尚礼.红、黄、绿三种颜色荧光质粒导入大肠杆菌中的稳定性表达[J].甘肃农业大学学报,2018,53(3):193-198.
[5] 杨晓玫,周彤,阿芸,等.苜蓿根瘤菌GFP荧光标记株的构建及对结瘤固氮的影响[J].草原与草坪,2018,38(1):44-56.
[6] Tan G Y.Genetic variation for acetylene reduction rate and other character in alfalfa[J].Crop Science,1981,21:485-488.
[7] 张虎天,郭丽琢,柴强,等.接种根瘤菌对豌豆/玉米体系根际细菌数量及氮营养的影响[J].甘肃农业大学学报,2011,46(1):30-33.
[8] 陈力玉,张淑卿,李剑锋,等.接种荧光标记根瘤菌对苜蓿幼苗生长的影响[J].草原与草坪,2013,33(6):1-8.
[9] 袁全,杨梦华,郑会明,等.中慢生型华癸根瘤菌Mesorhizobiumhuakuii AS9 自体诱导物合成酶基因的筛选及其功能分析[J].土壤,2009,41(3):459-463.
[10] 殷爱华,韩素芬.豆科树种凝集素和根瘤菌胞外多糖结合反应与结瘤的关系[J].南京林业大学学报(自然科学版),2005,29(5):88-90.
[11] Hoagland D,Arnon D I.The water culture method for growing plants without soil[J].California Agricultural Experiment Station Bulletin,1938(1):1-39.
[12] 高永革,李黎,刘祥等.黄河滩区紫花苜蓿生产性能比较研究[J].草业科学,2008,25(7):59-64.
[13] Vertucci C W,Roosee,Crane J.The oretical basis of protocols foe seed storage optimummoisture contents for pea seeds stored at different temperature[J].Annalsof Botany,1994,74:531-540.
[14] 李剑峰,师尚礼,张淑卿.环境酸度对紫花苜蓿早期生长和生理的影响[J].草业学报,2010,19(2):47-54.
[15] Hara S,Hashidoko Y,Desyatkin R V,et al.High rate of N2 fixation by east Siberian cryophilic soil bacteria as determined by measuring acetylene reduction in nitrogen poor medium solid ified with gellan gum[J].Appliedand Environmental Microbiology,2009,75:2811-2819.
[16] 谭志远,彭桂香,徐培智,等.普通野生稻(Oryzarufipogen)内生固氮菌多样性及高固氮酶活性[J] .科学通报,2009,54:1885-1893.
[17] 陈力玉.基于三亲本杂交的荧光标记根瘤菌的构建及其稳定性检测研究[D].兰州:甘肃农业大学硕士论文,2013.
[18] 邹琦.植物生理生化实验指导[M].北京:中国农业出版社,1995.
[19] Muhammad Z,AsifT,Ata C Z,et al.Weed-crop competition effects on growth and yield of sugarcane planted using two methods[J].Pakistan Journal of Botany,2010,42(2):815-823.
[20] 霍尔特(著).刘复今(译).伯杰鉴定手册[M](第八版).济南:山东大学出版社,1988:93-94.
[21] 张淑卿.根瘤菌在苜蓿植株体内的运移及影响因素[D].兰州:甘肃农业大学,2012.
[22] 陈翠翠,马元武,冯永君,等.MADS-box 家族蛋白在植物开花、结实及根瘤形成中的多功能调节作用[J].华北农学报,2008,23(S2):80-83.
[23] DudejaSS.PersistenceofDradyrhizobiumSP.(caianus)inasandyloam[J].SoilBoilBiochem,1989,21(5):709-713.
[24] 凌瑶,张小平,周俊初,等.用荧光酶基因(cfp)标记法研究菜豆根瘤菌的竞争性和有效性[J].中国土壤与肥料,2008(1):73-77.
[25] 韩颖,赵寿经,杨瑜,等,玉米陈化关键酶基因Zmlox-2的RNA干扰载体的构建[J].华南理工大学学报(自然科学版),2016,44(3):136-140.
[26] 董伟欣,张月辰,张迎迎.DsRED红色荧光蛋白植物表达载体的构建和表达[J].河北农业大学学报,2010,5(3):33.
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