转TaSCL14基因小麦的遗传稳定性及农艺性状分析
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摘要
为了获得农艺性状优良的转TaSCL14基因小麦品系,以普通小麦品种小偃39为受体获得的转TaSCL14基因小麦T1～T3代株(系)为材料,对其中TaSCL14基因的遗传稳定性和主要的农艺特性进行分析。结果表明,转TaSCL14基因小麦的T1和T2代植株的阳性率分别达到55.9%和85.6%,穗粒数、冬前分蘖与对照存在显著差异;T3代4个转基因株系3-4、3-7、3-8和3-11的阳性率达到90%以上,且TaSCL14基因在各株系中的表达水平都高于对照。与野生型相比,转基因小麦T3代部分株系的冬前分蘖、冬后分蘖、有效分蘖和小穗数均较野生型呈显著或极显著提高,但千粒重都较野生型有所降低,其中株系3-4和3-7降低幅度较大,分别达到显著和极显著水平。以上结果说明,TaSCL14基因的过表达能够影响转基因小麦的部分农艺特性,并且在株(系)间有差异。
In order to obtain the optimal TaSCL14 transgenic lines with great agronomic traits,T1,T2,T3 of common wheat cultivar Xiaoyan 39 transformed with TaSCL14 were used as materials in this experiment.The positive transgenic plants were detected and screened by PCR,and the genetic stability characteristics and major agronomic traits of TaSCL14 gene in transgenic lines T1,T2 and T3were analyzed.The results showed that the positive rate of T1 and T2 plants of TaSCL14 transgenic wheat reached to 55.9% and 85.6%,respectively,and there were significant differences in the number of grains per spike,tillers before winter compared with those of control.And the positive rate of the four transgenic lines of the T3 generation transgenic lines reached to more than 90%,indicating that TaSCL14 gene can be stably inherited to offspring and overexpressed in each strain.The T3 lines showed significant differences from the control in tillering before winter,tillers in winter,effective tillers,grain number per spike and 1000-grain weight.Thousand grain weight of lines 3-4and 3-7was significant lower than that of the control.It was indicated that overexpression of TaSCL14 affected some agronomic traits in transgenic wheat and showed differences between strains(lines).
In order to obtain the optimal TaSCL14 transgenic lines with great agronomic traits,T1,T2,T3 of common wheat cultivar Xiaoyan 39 transformed with TaSCL14 were used as materials in this experiment.The positive transgenic plants were detected and screened by PCR,and the genetic stability characteristics and major agronomic traits of TaSCL14 gene in transgenic lines T1,T2 and T3were analyzed.The results showed that the positive rate of T1 and T2 plants of TaSCL14 transgenic wheat reached to 55.9% and 85.6%,respectively,and there were significant differences in the number of grains per spike,tillers before winter compared with those of control.And the positive rate of the four transgenic lines of the T3 generation transgenic lines reached to more than 90%,indicating that TaSCL14 gene can be stably inherited to offspring and overexpressed in each strain.The T3 lines showed significant differences from the control in tillering before winter,tillers in winter,effective tillers,grain number per spike and 1000-grain weight.Thousand grain weight of lines 3-4and 3-7was significant lower than that of the control.It was indicated that overexpression of TaSCL14 affected some agronomic traits in transgenic wheat and showed differences between strains(lines).
引文
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[16]陈立勇,陈丽华,柴丽娟,等.葡萄VvSCL9基因和启动子的克隆及初步研究[J].中国农业大学学报,2015,20(2):101.CHEN L Y,CHEN H L,CHAI L J,et al.Isolation and primary function study of VvSCL9 gene and its promoter[J].Journal of China Agricultural University,2015,20(2):101.
[17]MA H S,LIANG D,SHUAI P,et al.The salt-and droughtinducible poplar GRAS protein SCL7 confers salt and drought tolerance in Arabidopsis thaliana[J].Journal of Experimental Botany,2010,61(14):4011.
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[20]CZIKKEL B E,MAXWELL D P.NtGRAS1,a novel stressinduced member of the GRAS family in tobacco,localizes to the nucleus[J].Journal of Plant Physiology,2007,164(9):1220.
[21]DI L L,WYSOCKA-DILLE J,MALAMY J E,et al.The SCARECROW gene regulates an asymmetric cell division that is essential for generating the radial organization of the Arabidopsis root[J].Cell,1996,86(3):423.
[22]陈坤梅.小麦高光效相关基因功能分析及TaSCL14基因克隆与功能验证[D].杨凌:西北农林科技大学,2015:92.CHEN K M.Analysis of high photosynthetic efficiency genomic function in wheat and cloning and function verification of TaSCL14 gene[D].Yangling:Northwest A&F University,2015:92.
[23]叶兴国,徐惠君,杜丽璞,等.小麦规模化转基因技术体系构建及其应用[J].中国农业科学,2014,47(21):4155.YE X G,XU H J,DU L P,et al.Establishment and application of Large-Scale transformation systems in wheat[J].Scientia Agricultura Sinica,2014,47(21):4155.
[24]王军卫,张改生,张晓东.脱水应答转录因子CBF1的克隆与转基因小麦的分子检测[J].西北植物学报.2008,28(2):2223.WANG J W,ZHANG G S,ZHANG X D.Cloning of CBF1transcriptional factor gene and molecular analysis in transgenic wheat[J].Acta Botanica Boreantalia Sinica,2008,28(2):2223.
[25]李陈建,买寅生,付彦博,等.转sGNA基因小麦农艺性状评价与抗蚜效果分析[J].新疆农业科学,2015,52(3):503.LI C J,MAI Y S,FU Y B,et al.A preliminary analysis on agronomic aerformance and aphid-resistance of transgenic wheat with sGNA gene[J].Xinjiang Agricultural Sciences,2015,52(3):503.
[26]闫海生,雷闯,马凌霄,等.转OsCYP2基因水稻后代的遗传分析及农艺性状观察[J].湖北农业科学,2013,52(7):1510.YAN H S,LEI C,MA L X,et al.Genetic analysis and agronomic characters observation on transgenic OsCYP2 rice[J].Hubei Agricultural Sciences,2013,52(7):1510.
[27]秦保平,杨敏,王振林,等.外源Hpal10-42基因导入对小麦农艺性状和籽粒品质的影响[J].山东农业科学,2013,45(9):19.QIN B P,YANG M,WANG Z L,et al.Effects of foreign Hpal10-42 gene introduction on agronomic traits and kernel quality of wheat[J].Shandong Agricultural Sciences,2013,45(9):19.
[28]王瑞霞,高庆荣,崔德才,等.转反义PLDγ基因小麦的分子检测及农艺性状分析[J].作物学报,2005,31(10):1354.WANG R X,GAO Q R,CUI D C,et al.Molecular detection of transformed wheat with Anti-PLDγgene and its agronomic characters[J].Acta Agronomica Sinica,2005,31(10):1354.
[29]BREGITZER P,HALBERT S E,LEMAUX P G.Somaclonal variation in the progeny of transgenic barley[J].Theoretical and Applied Genetics,1998,96(3-4):421.
[30]LUEHRSEN K R,WALBOT V.Insertion of Mu1elements in the first intron of the Adh1-Sgene of maize results in novel RNA processing events[J].The Plant Cell,1990,2(12):1225.
[2]VASIL V,CASTILLO A M,FROMM M E,et al.Herbicide resistantfertile transgenic wheat plants obtained by microprojectile bombardment of regenerable embryogenic callus[J].Nature Biotechnology,1992(10):667.
[3]徐琼芳,田芳,陈孝,等.转基因抗虫小麦中sgna基因的遗传分析及抗虫性鉴定[J].作物学报,2004,30(5):475.XU Q F,TIAN F,CHEN X,et al.Inheritance of sgna gene and insect-resistant activity in transgenic wheat[J].Acta Agronomica Sinica,2004,30(5):475.
[4]茹京娜,于太飞,陈隽,等.小麦锌指转录因子TaDi19A对低温的响应及其互作蛋白的筛选[J].中国农业科学,2017,50(13):2411.RU J N,YU T F,CHEN J,et al.Response of wheat Zinc-Finger transcription factor TaDi19Ato cold and its screening of interacting proteins[J].Scientia Agricultura Sinica,2017,50(13):2411.
[5]陈凡国,夏光敏.新型HMW-GS基因H1By16改良转基因小麦面粉品质[J].植物生理学报,2013,49(7):664.CHEN F G,XIA G M.Transformation of wheat with a novel high-molecular-weight glutenin subunit gene H1By16 improving functional properties of wheat flour[J].Plant Physiology Journal,2013,49(7):664.
[6]张彬,丁在松,张桂芳,等.根癌农杆菌介导获得稗草Ecppc转基因小麦的研究[J].作物学报,2007,33(3):356.ZHANG B,DING Z S,ZHANG G F,et al.Introduction of phosphoenolpyruvate carboxylase gene from echinochloa crusgalli into wheat mediated by Agrobacterium tumefaciens[J].Acta Agronomica Sinica,2007,33(3):356.
[7]喻修道,徐兆师,陈明,等.小麦转基因技术研究及其应用[J].中国农业科学,2010,43(8):1539.YU X D,XU Z S,CHEN M,et al.The progress and application of wheat transformation technology[J].Scientia Agricultura Sinica,2010,43(8):1539.
[8]KU M S,AGARIE S,NOMURA M,et al.High-level expression of maize phosphoenolpyruvate carboxylase in transgenic rice plants[J].Nature Biotechnology,1999,17(1):76.
[9]张建红,许为钢,王会伟,等.转玉米C4型高光效pepc基因小麦的分子特征及光合特性研究[J].麦类作物学报,2012,32(6):1043.ZHANG J H,XU W G,WANG H W,et al.Molecular characteristics and photosynthetic property of the transgenic wheat expressing a maize C4-type pepc gene[J].Journal of Triticeae Crops,2012,32(6):1043.
[10]张艳,满为群,南相日,等.高粱C4型pepc基因转入大豆可改善大豆光合特性[J].分子植物育种,2015,13(2):294.ZHANG Y,MAN W Q,NAN X Y,et al.Sorghum C4-Specific pepc gene transformed into soybean can improve the photosynthetic characteristics of soybean[J].Molecular Plant Breeding,2015,13(2):294.
[11]张建福,Datta Swapan K,王国英,等.玉米高光效基因PPDK在籼稻IR64中的整合及其与光合作用相关的特性分析[J].分子植物育种.2006,4(6):797.ZHANG J F,Swapan K.Datta,WANG G Y,et al.Integration of C4-specific PPDKgene of maize to C3rice IR64and its photosynthesis characteristics analysis[J].Molecular Plant Breeding,2006,4(6):797.
[12]Pysh L D,Wysocka-Diller J W,Camilleri C,et al.The GRAS gene family in Arabidopsis:sequence characterization and basic expression analysis of the SCARECROW-LIKE genes[J].The Plant Journal,2010,18(1):111.
[13]TIAN C G,WAN P,SUN S H,et al.Genome-wide analysis of the GRASgene family in rice and Arabidopsis[J].Plant Molecular Biology,2004,54(4):519.
[14]李桂英,田玉富,杨成君.植物GRAS家族转录因子的研究现状[J].安徽农业科学,2014(14):4207.LI G Y,TIAN Y F,YANG C J.Research situation of GRAS family trarscription factor in plants[J].Journal of Anhui Agricultural Sciences,2014(14):4207.
[15]XU K,CHEN S,LI T,et al.OsGRAS23,a rice GRAS transcription factor gene,is involved in drought stress response through regulating expression of stress-responsive genes[J].BMC Plant Biology,2015,7(13):15.
[16]陈立勇,陈丽华,柴丽娟,等.葡萄VvSCL9基因和启动子的克隆及初步研究[J].中国农业大学学报,2015,20(2):101.CHEN L Y,CHEN H L,CHAI L J,et al.Isolation and primary function study of VvSCL9 gene and its promoter[J].Journal of China Agricultural University,2015,20(2):101.
[17]MA H S,LIANG D,SHUAI P,et al.The salt-and droughtinducible poplar GRAS protein SCL7 confers salt and drought tolerance in Arabidopsis thaliana[J].Journal of Experimental Botany,2010,61(14):4011.
[18]石瑞,曹诣斌,陈文荣,等.佛手GRAS基因的克隆及表达分析[J].浙江师范大学学报(自然科学版),2011,34(4):446.SHI R,CAO Y B,CHEN W R,et al.On cDNA cloning and expression analysis of GRAS gene in fingered citron[J].Journal of Zhejiang Normal University(Natural Sciences),2011,34(4):446.
[19]李阿英,刘洪,李晓颖,等.枳两个GRAS基因cDNA全长的克隆及其亚细胞定位分析[J].基因组学与应用生物学,2012,31(3):240.LI A Y,LIU H,LI X Y,et al.Cloning and subcellular localization analysis of two GRASgenes from Poncirus trifoliata[J].Genomics and Applied Biology,2012,31(3):240.
[20]CZIKKEL B E,MAXWELL D P.NtGRAS1,a novel stressinduced member of the GRAS family in tobacco,localizes to the nucleus[J].Journal of Plant Physiology,2007,164(9):1220.
[21]DI L L,WYSOCKA-DILLE J,MALAMY J E,et al.The SCARECROW gene regulates an asymmetric cell division that is essential for generating the radial organization of the Arabidopsis root[J].Cell,1996,86(3):423.
[22]陈坤梅.小麦高光效相关基因功能分析及TaSCL14基因克隆与功能验证[D].杨凌:西北农林科技大学,2015:92.CHEN K M.Analysis of high photosynthetic efficiency genomic function in wheat and cloning and function verification of TaSCL14 gene[D].Yangling:Northwest A&F University,2015:92.
[23]叶兴国,徐惠君,杜丽璞,等.小麦规模化转基因技术体系构建及其应用[J].中国农业科学,2014,47(21):4155.YE X G,XU H J,DU L P,et al.Establishment and application of Large-Scale transformation systems in wheat[J].Scientia Agricultura Sinica,2014,47(21):4155.
[24]王军卫,张改生,张晓东.脱水应答转录因子CBF1的克隆与转基因小麦的分子检测[J].西北植物学报.2008,28(2):2223.WANG J W,ZHANG G S,ZHANG X D.Cloning of CBF1transcriptional factor gene and molecular analysis in transgenic wheat[J].Acta Botanica Boreantalia Sinica,2008,28(2):2223.
[25]李陈建,买寅生,付彦博,等.转sGNA基因小麦农艺性状评价与抗蚜效果分析[J].新疆农业科学,2015,52(3):503.LI C J,MAI Y S,FU Y B,et al.A preliminary analysis on agronomic aerformance and aphid-resistance of transgenic wheat with sGNA gene[J].Xinjiang Agricultural Sciences,2015,52(3):503.
[26]闫海生,雷闯,马凌霄,等.转OsCYP2基因水稻后代的遗传分析及农艺性状观察[J].湖北农业科学,2013,52(7):1510.YAN H S,LEI C,MA L X,et al.Genetic analysis and agronomic characters observation on transgenic OsCYP2 rice[J].Hubei Agricultural Sciences,2013,52(7):1510.
[27]秦保平,杨敏,王振林,等.外源Hpal10-42基因导入对小麦农艺性状和籽粒品质的影响[J].山东农业科学,2013,45(9):19.QIN B P,YANG M,WANG Z L,et al.Effects of foreign Hpal10-42 gene introduction on agronomic traits and kernel quality of wheat[J].Shandong Agricultural Sciences,2013,45(9):19.
[28]王瑞霞,高庆荣,崔德才,等.转反义PLDγ基因小麦的分子检测及农艺性状分析[J].作物学报,2005,31(10):1354.WANG R X,GAO Q R,CUI D C,et al.Molecular detection of transformed wheat with Anti-PLDγgene and its agronomic characters[J].Acta Agronomica Sinica,2005,31(10):1354.
[29]BREGITZER P,HALBERT S E,LEMAUX P G.Somaclonal variation in the progeny of transgenic barley[J].Theoretical and Applied Genetics,1998,96(3-4):421.
[30]LUEHRSEN K R,WALBOT V.Insertion of Mu1elements in the first intron of the Adh1-Sgene of maize results in novel RNA processing events[J].The Plant Cell,1990,2(12):1225.