转AtSnRK2C基因杨树田间试验及生物安全性分析
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摘要
为探讨转AtSnRK2C基因"南林895"杨对周围生态环境的影响,本研究从转基因植株树高、外源基因稳定性、外源基因对土壤微生物数量的影响、外源基因水平转移以及转基因植株叶片化感作用效应等5个方面,对转AtSnRK2C基因"南林895"杨进行了田间试验及生物安全性分析。分析结果显示,两个转AtSnRK2C基因"南林895"杨株系树高的平均值均高于非转基因杨树,但树高差异不显著;在转AtSnRK2C基因"南林895"杨进入田间试验一年后外源基因仍稳定表达;转AtSnRK2C基因"南林895"杨对土壤微生物数量无显著性影响;外源基因AtSnRK2C未发生水平转移;转AtSnRK2C基因"南林895"杨叶片对生菜种子的生长无显著性影响。本研究为转基因杨树抗旱耐盐新品种培育及应用提供了理论依据。
In order to explore the effects of transgenic "Nanlin895" poplar with AtSnRK2 C gene on the surrounding ecological environment, field trials and the biosafety assessment of transgenic "Nanlin 895" poplar with AtSnRK2 C gene were carried out from five aspects in this study: the height of transgenic plants, the stability of exogenous genes, the influence of exogenous genes on soil microbial populations, the horizontal transfer of exogenous genes, and the allelopathy effects of transgenic plants. The results showed that the average tree height of the two transgenic lines was higher than that of the non-transgenic plants, but the difference did not reach a significant difference. The expression of exogenous genes was still stable one year after the transfer of AtSnRK2 C gene "Nanlin895" poplar into the field trial. There was no significant effect on the quantity of soil microbes and no horizontal transfer of exogenous genes. The leaves of transgenic plants had no significant effect on lettuce seed growth. The results of this study would provide theoretical basis for breeding and application of drought-tolerance and salt-tolerance varieties of transgenic poplar.
In order to explore the effects of transgenic "Nanlin895" poplar with AtSnRK2 C gene on the surrounding ecological environment, field trials and the biosafety assessment of transgenic "Nanlin 895" poplar with AtSnRK2 C gene were carried out from five aspects in this study: the height of transgenic plants, the stability of exogenous genes, the influence of exogenous genes on soil microbial populations, the horizontal transfer of exogenous genes, and the allelopathy effects of transgenic plants. The results showed that the average tree height of the two transgenic lines was higher than that of the non-transgenic plants, but the difference did not reach a significant difference. The expression of exogenous genes was still stable one year after the transfer of AtSnRK2 C gene "Nanlin895" poplar into the field trial. There was no significant effect on the quantity of soil microbes and no horizontal transfer of exogenous genes. The leaves of transgenic plants had no significant effect on lettuce seed growth. The results of this study would provide theoretical basis for breeding and application of drought-tolerance and salt-tolerance varieties of transgenic poplar.
引文
Brodersen P.,and Voinnet O.,2006,The diversity of RNA silencing pathways in plants,Trends Genet.,22(5):268-280
Chen X.W.,Lin S.,You M.S.,Yang G.,and Wang F.,2011,Effects of transgenic rice on the structure and function of soil microbial community,Shengwu Anquan Xuebao(Journal of Biosafety),20(2):151-159(陈晓雯,林胜,尤民生,杨广,王锋,2011,转基因水稻对土壤微生物群落结构及功能的影响,生物安全学报,20(2):151-159)
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Fujii Y.,Parvez S.S.,Parvez M.M.,Ohmae Y.,and Lida O.,2003,Screening of 239 medicinal plant species for allelopathic activity using the sandwich method,Weed Biol.Manag.,3(4):233-241
Kiran S.,Pooja B.M.,and Trevor T.,2005,Genetic transformation technology:status and problems,In Vitro Cell Dev-Pl.,41(2):102-112
Kohli A.,Twyman R.M.,Abranches R.,Wegel E.,Stoger E.,and Christou P.,2003,Transgene integration,organization and interaction in plants,Plant Mol.Biol.,52(2):247-258
Li H.F.,Liu Y.,Kan Y.,and Wang Q.Y.,2014,Influence of transgenic Populus simonii×P.nigra on soil microbial community,Nanjing Linye Daxue Xuebao(Journal of Nanjing Forestry University(Natural Science Edition)),38(2):75-80(李海峰,刘岩,康颖,王秋玉,2014,转基因小黑杨对土壤微生物群落结构的影响,南京林业大学学报(自然科学版),38(2):75-80)
Li X.,Xue Q.H.,Fan J.F.,Zhou Y.X.,and Gao J.S.,2011,Effects of one-year-old Bt transgenic Populus nigra on soil microorganism,Xibei Linyueyuan Xuebao(Journal of Northwest Forestry University),26(4):123-127(李霞,薛泉宏,樊军锋,周永学,高建设,2011,一年生转Bt基因欧洲黑杨对土壤微生物的影响,西北林学院学报,26(4):123-127)
Mao X.,Zhang H.,Tian S.,Chang X.,and Jing R.,2010,Ta S-n RK2.4,an SNF1-type serine/threonine protein kinase of wheat(Triticum aestivum L.),confers enhanced multistress tolerance in Arabidopsis,J.Exp.Bot.,61(3):683-696
Oguchi T.,Kashimura Y.,Mimura M.,Yu X.,Matsunaga E.,Nanto K.,Shimada T.,Kikuchi A.,and Watanabe K.N.,2014,A multi-year assessment of the environmental impact of transgenic Eucalyptus trees harboring a bacterial choline oxidase gene on biomass,precinct vegetation and the microbial community,Transgenic Res.,23(5):767-777
Perry J.N.,Ter Braak C.J.,Dixon P.M.,Duan J.J.,Hails R.S.,Huesken A.,Lavielle M.,Marvier M.,Scardi M.,Schmidt K.,Tothmeresz B.,Schaarschmidt F.,and van der Voet H.,2009,A statistical aspects of environmental risk assessment of GM plants for effects on non-target organisms,Environ.Biosafety Res.,8(2):65-78
Prescott V.E.,and Hogan S.P.,2006,Genetically modified plants and food hypersensitivity diseases:usage and implications of experimental models for risk assessment,Pharmacol.Ther.,111(2):374-383
Richter B.,and Smalla K.,2007,Screening of rhizosphere and soil bacteria for transformability,Environ.Biosafety Res.,6(1):91-99
Song X.Q.,and Zhuge Q.,2016,Progress in a plant gene family of sucrose non-fermentiing 1-related protein kinase 2,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),14(4):870-877(宋雪晴,诸葛强,2016,植物Sn RK2基因家族研究进展,分子植物育种,14(4):870-877)
Song X.Q.,Yu X.,Hori C.,Demura T.,Ohtani M.,and Zhuge Q.,2016,Heterologous overexpression of poplar Sn RK2 genes enhanced salt stress tolerance in Arabidopsis thaliana,Front.Plant Sci.,7:612
Teng Y.,and Huang C.Y.,2002,Ecological effect of heavy metals on soil microbes and research advances on the mechanisms of bioremediation,Huanjing Yu Turang(Soil&Environmental Sciences),11(1):85-89(滕应,黄昌勇,2002,重金属污染土壤的微生物生态效应及其修复研究进展,土壤与环境,11(1):85-89)
Umezawa T.,Yoshida R.,Maruyama K.,Yamaguchi-Shinozaki K.,and Shinozaki K.,2004,SRK2C,a SNF1-related protein kinase 2,improves drought tolerance by controlling stressresponsive gene expression in Arabidopsis thaliana,Proc.Natl.Acad.Sci.USA,101(49):17306-17311
Ying S.,Zhang D.F.,Li H.Y.,Liu Y.H.,Shi Y.S.,Song Y.C.,Wang T.Y.,and Li Y.,2011,Cloning and characterization of a maize Sn RK2 protein kinase gene confers enhanced salt tolerance in transgenic Arabidopsis,Plant Cell Rep.,30(9):1683-1699
Yu X.,Kikuchi A.,Shimazaki T.,Yamada A.,Ozeki Y.,Matsunaga E.,Ebinuma H.,and Watanabe K.N.,2013,Assessment of the salt tolerance and environmental biosafety of Eucalyptus camaldulensis harboring a mangrin transgene,J.Plant Res.,126(1):141-150
Zhang Y.,Guo T.B.,Pan H.X.,Huang M.R.,Wang M.X.,and Zhuge Q.,2012,Analysis on insecticidal activity of Bt transgenic Populus deltoides×P.euramericana cv'Nanlin895'and its effects on soil microorganism,Linye Kexue Yanjiu(Journal of Forestry Research),25(3):346-350(张雁,郭同斌,潘惠新,黄敏仁,王明庥,诸葛强,2012,转Bt基因南林895杨抗虫性及对土壤微生物影响分析,林业科学研究,25(3):346-350)
Zhu W.X.,Ding C.J.,Zhang W.X.,Zhang B.Y.,Huang Q.J.,Chu Y.G.,and Su X.H.,2017,Exogenous gene transformation of8-year-old multi-gene transgenic Populus×euramericana'Guariento'and its influence on soil microbial quantity,Linye Kexue Yanjiu(Journal of Forestry Research),30(2):349-353(朱文旭,丁昌俊,张伟溪,张冰玉,黄秦军,褚延广,苏晓华,2017,8年生转基因库安托杨外源基因转移及对土壤微生物数量影响的检测,林业科学研究,30(2):349-353)
Zhu Z.,and Liu X.,2000,Genetically modified crops-frankenstein or savior,Nongye Shengwu Jishu Xuebao(Journal of Agricultural Biotechnology),8(1):1-5(朱祯,刘翔,2000,转基因作物-恶魔还是救星,农业生物技术学报,8(1):1-5)
Chen X.W.,Lin S.,You M.S.,Yang G.,and Wang F.,2011,Effects of transgenic rice on the structure and function of soil microbial community,Shengwu Anquan Xuebao(Journal of Biosafety),20(2):151-159(陈晓雯,林胜,尤民生,杨广,王锋,2011,转基因水稻对土壤微生物群落结构及功能的影响,生物安全学报,20(2):151-159)
Diedhiou C.J.,Popova O.V.,Dietz K.J.,and Golldack D.,2008,The SNF1-type serine-threonine protein kinase SAPK4 regulates stress-responsive gene expression in rice,BMC Plant Biol.,8(1):49
Fujii Y.,Parvez S.S.,Parvez M.M.,Ohmae Y.,and Lida O.,2003,Screening of 239 medicinal plant species for allelopathic activity using the sandwich method,Weed Biol.Manag.,3(4):233-241
Kiran S.,Pooja B.M.,and Trevor T.,2005,Genetic transformation technology:status and problems,In Vitro Cell Dev-Pl.,41(2):102-112
Kohli A.,Twyman R.M.,Abranches R.,Wegel E.,Stoger E.,and Christou P.,2003,Transgene integration,organization and interaction in plants,Plant Mol.Biol.,52(2):247-258
Li H.F.,Liu Y.,Kan Y.,and Wang Q.Y.,2014,Influence of transgenic Populus simonii×P.nigra on soil microbial community,Nanjing Linye Daxue Xuebao(Journal of Nanjing Forestry University(Natural Science Edition)),38(2):75-80(李海峰,刘岩,康颖,王秋玉,2014,转基因小黑杨对土壤微生物群落结构的影响,南京林业大学学报(自然科学版),38(2):75-80)
Li X.,Xue Q.H.,Fan J.F.,Zhou Y.X.,and Gao J.S.,2011,Effects of one-year-old Bt transgenic Populus nigra on soil microorganism,Xibei Linyueyuan Xuebao(Journal of Northwest Forestry University),26(4):123-127(李霞,薛泉宏,樊军锋,周永学,高建设,2011,一年生转Bt基因欧洲黑杨对土壤微生物的影响,西北林学院学报,26(4):123-127)
Mao X.,Zhang H.,Tian S.,Chang X.,and Jing R.,2010,Ta S-n RK2.4,an SNF1-type serine/threonine protein kinase of wheat(Triticum aestivum L.),confers enhanced multistress tolerance in Arabidopsis,J.Exp.Bot.,61(3):683-696
Oguchi T.,Kashimura Y.,Mimura M.,Yu X.,Matsunaga E.,Nanto K.,Shimada T.,Kikuchi A.,and Watanabe K.N.,2014,A multi-year assessment of the environmental impact of transgenic Eucalyptus trees harboring a bacterial choline oxidase gene on biomass,precinct vegetation and the microbial community,Transgenic Res.,23(5):767-777
Perry J.N.,Ter Braak C.J.,Dixon P.M.,Duan J.J.,Hails R.S.,Huesken A.,Lavielle M.,Marvier M.,Scardi M.,Schmidt K.,Tothmeresz B.,Schaarschmidt F.,and van der Voet H.,2009,A statistical aspects of environmental risk assessment of GM plants for effects on non-target organisms,Environ.Biosafety Res.,8(2):65-78
Prescott V.E.,and Hogan S.P.,2006,Genetically modified plants and food hypersensitivity diseases:usage and implications of experimental models for risk assessment,Pharmacol.Ther.,111(2):374-383
Richter B.,and Smalla K.,2007,Screening of rhizosphere and soil bacteria for transformability,Environ.Biosafety Res.,6(1):91-99
Song X.Q.,and Zhuge Q.,2016,Progress in a plant gene family of sucrose non-fermentiing 1-related protein kinase 2,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),14(4):870-877(宋雪晴,诸葛强,2016,植物Sn RK2基因家族研究进展,分子植物育种,14(4):870-877)
Song X.Q.,Yu X.,Hori C.,Demura T.,Ohtani M.,and Zhuge Q.,2016,Heterologous overexpression of poplar Sn RK2 genes enhanced salt stress tolerance in Arabidopsis thaliana,Front.Plant Sci.,7:612
Teng Y.,and Huang C.Y.,2002,Ecological effect of heavy metals on soil microbes and research advances on the mechanisms of bioremediation,Huanjing Yu Turang(Soil&Environmental Sciences),11(1):85-89(滕应,黄昌勇,2002,重金属污染土壤的微生物生态效应及其修复研究进展,土壤与环境,11(1):85-89)
Umezawa T.,Yoshida R.,Maruyama K.,Yamaguchi-Shinozaki K.,and Shinozaki K.,2004,SRK2C,a SNF1-related protein kinase 2,improves drought tolerance by controlling stressresponsive gene expression in Arabidopsis thaliana,Proc.Natl.Acad.Sci.USA,101(49):17306-17311
Ying S.,Zhang D.F.,Li H.Y.,Liu Y.H.,Shi Y.S.,Song Y.C.,Wang T.Y.,and Li Y.,2011,Cloning and characterization of a maize Sn RK2 protein kinase gene confers enhanced salt tolerance in transgenic Arabidopsis,Plant Cell Rep.,30(9):1683-1699
Yu X.,Kikuchi A.,Shimazaki T.,Yamada A.,Ozeki Y.,Matsunaga E.,Ebinuma H.,and Watanabe K.N.,2013,Assessment of the salt tolerance and environmental biosafety of Eucalyptus camaldulensis harboring a mangrin transgene,J.Plant Res.,126(1):141-150
Zhang Y.,Guo T.B.,Pan H.X.,Huang M.R.,Wang M.X.,and Zhuge Q.,2012,Analysis on insecticidal activity of Bt transgenic Populus deltoides×P.euramericana cv'Nanlin895'and its effects on soil microorganism,Linye Kexue Yanjiu(Journal of Forestry Research),25(3):346-350(张雁,郭同斌,潘惠新,黄敏仁,王明庥,诸葛强,2012,转Bt基因南林895杨抗虫性及对土壤微生物影响分析,林业科学研究,25(3):346-350)
Zhu W.X.,Ding C.J.,Zhang W.X.,Zhang B.Y.,Huang Q.J.,Chu Y.G.,and Su X.H.,2017,Exogenous gene transformation of8-year-old multi-gene transgenic Populus×euramericana'Guariento'and its influence on soil microbial quantity,Linye Kexue Yanjiu(Journal of Forestry Research),30(2):349-353(朱文旭,丁昌俊,张伟溪,张冰玉,黄秦军,褚延广,苏晓华,2017,8年生转基因库安托杨外源基因转移及对土壤微生物数量影响的检测,林业科学研究,30(2):349-353)
Zhu Z.,and Liu X.,2000,Genetically modified crops-frankenstein or savior,Nongye Shengwu Jishu Xuebao(Journal of Agricultural Biotechnology),8(1):1-5(朱祯,刘翔,2000,转基因作物-恶魔还是救星,农业生物技术学报,8(1):1-5)
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