四川小麦优良受体基因型筛选及农杆菌转化因素优化
|
摘要
为了丰富栽培小麦转化受体基因型,优化农杆菌转化小麦幼胚的技术体系,本研究评价了21份四川优良小麦品种(系)的幼胚再生能力,并以筛选到的最佳受体为材料,利用正交设计研究了预培养时间、菌液浓度、侵染时间、抑菌剂浓度、共培养方式和农杆菌菌株等易互作的6个因素对幼胚抗性愈伤再生率的影响,并探讨了筛选剂卡那霉素(Kanamycin)的适宜浓度。结果表明,21份材料中‘5157’、‘川农16’、‘R364’、‘川麦42’‘、内麦8号’的绿苗率超过了30%,可作为优良转基因受体加以利用。以‘5157’为受体材料建立农杆菌转化体系,正交试验表明菌液浓度和农杆菌菌株对幼胚抗性愈伤再生率的影响达到显著水平,而其他因素影响不显著。经优化后的遗传转化体系为:农杆菌菌株用C58C1,幼胚不经预培养,在菌液OD值为0.4的农杆菌中侵染40 min,共培养介质选用培养基,抑菌剂为200 mg/L羧卞青霉素(Carbenicillin),适宜的卡那霉素筛选浓度为50 mg/L。本研究为四川小麦遗传转化提供了5个候选基因型,为建立和优化小麦幼胚转化技术体系提供了理论依据,同时初步建立了农杆菌转化小麦幼胚的技术体系。
In order to enrich the genotype of wheat transformation receptor and optimize the technical system of wheat immature embryo transformation by Agrobacterium tumefaciens, the regeneration ability of immature embryos of 21 Sichuan wheat varieties(lines) was evaluated, and the optimal receptors were selected as materials.The pre-culture time, concentration of bacterial solution, infection time and bacteriostasis were studied by orthogonal design. The effects of six factors, such as concentration, co-culture mode and Agrobacterium strains, on the regeneration rate of immature embryo-resistant callus were studied. The suitable concentration of kanamycin was also discussed. Results indicated that the green plant regeneration rate of varieties(lines) '5157', 'Chuannong 16', 'R364', 'Chuanmai 42', and 'Neimai 8' were more than 30%, which could be used as superior genotypes in immature embryos culture and transformation. The transformation system of Agrobacterium tumefaciens was established with '5157' as acceptor material. The orthogonal test showed that the concentration of bacterial solution and Agrobacterium tumefaciens strain had significant effect on the regeneration rate of immature embryo resistant callus, but other factors had no significant effect. The optimized genetic transformation system was as follows:Agrobacterium strain C58 C1, immature embryo without pre-culture, Agrobacterium strain with OD value of 0.4 was infected for 40 minutes, co-culture medium was used, inhibitor was 200 mg/L carbenicillin, and the suitable concentration of kanamycin was 50 mg/L. This study provided five candidate genotypes for Sichuan wheat genetic transformation. Also, it provided theoretical references for establishment and optimization of the transformation system of wheat immature embryo, and preliminarily established the technical system of Agrobacterium mediated transformation of wheat immature embryos.
In order to enrich the genotype of wheat transformation receptor and optimize the technical system of wheat immature embryo transformation by Agrobacterium tumefaciens, the regeneration ability of immature embryos of 21 Sichuan wheat varieties(lines) was evaluated, and the optimal receptors were selected as materials.The pre-culture time, concentration of bacterial solution, infection time and bacteriostasis were studied by orthogonal design. The effects of six factors, such as concentration, co-culture mode and Agrobacterium strains, on the regeneration rate of immature embryo-resistant callus were studied. The suitable concentration of kanamycin was also discussed. Results indicated that the green plant regeneration rate of varieties(lines) '5157', 'Chuannong 16', 'R364', 'Chuanmai 42', and 'Neimai 8' were more than 30%, which could be used as superior genotypes in immature embryos culture and transformation. The transformation system of Agrobacterium tumefaciens was established with '5157' as acceptor material. The orthogonal test showed that the concentration of bacterial solution and Agrobacterium tumefaciens strain had significant effect on the regeneration rate of immature embryo resistant callus, but other factors had no significant effect. The optimized genetic transformation system was as follows:Agrobacterium strain C58 C1, immature embryo without pre-culture, Agrobacterium strain with OD value of 0.4 was infected for 40 minutes, co-culture medium was used, inhibitor was 200 mg/L carbenicillin, and the suitable concentration of kanamycin was 50 mg/L. This study provided five candidate genotypes for Sichuan wheat genetic transformation. Also, it provided theoretical references for establishment and optimization of the transformation system of wheat immature embryo, and preliminarily established the technical system of Agrobacterium mediated transformation of wheat immature embryos.
引文
Cheng M.,Fry J.E.,Pang S.Z.,Zhou H.P.,Hironaka C.M.,Duncan D.R.,Conner T.W.,and Wan Y.C.,1997,Genetic transformation of wheat mediated by Agrobacterium tumefaciens,Plant Physiol.,115(3):971-980
Ding L.,Li S.,Gao J.,Wang Y.,Yang G.,and He G.,2009,Optimization of Agrobacterium-mediated transformation conditions in mature embryos of elite wheat,Mol.Biol.Rep.,36(1):29-36
Du W.P.,Yu G.R.,Song J.,Xu L.Y.,Zhang L.,and Jiang Y.,2012,Effects of genotypes and explants on in vitro culture of wheat,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),10(3):290-297(杜文平,余桂容,宋军,徐利远,张莲,蒋云,2012,基因型和外植体对小麦离体培养的影响,分子植物育种,10(3):290-297)
Gao J.S.,Xu L.,Chen W.,Shen Z.L.,Cai Y.P.,and Lin Y.,2017,Agrobacterium-mediated genetic transformation of stem apexes of brown fiber cotton Zongcaixuan 1,Henongxue Bao(Journal of Nuclear Agricultural Sciences),31(10):1881-1888(高俊山,许磊,陈文,沈知临,蔡永萍,林毅,2017,农杆菌介导陆地棉棕彩选1号茎尖遗传转化,核农学报,31(10):1881-1888)
Godwin I.,Todd G.,Fordlloyd B.,and Newbury H.J.,1991,The effects of acetosyringone and p H on Agrobacterium-mediated transformation vary according to plant species,Plant Cell Rep.,9(12):671-675
He Y.,Jones H.D.,Chen S.,Chen X.M.,Wang D.W.,Li K.X.,Wang D.S.,and Xia L.Q.,2010,Agrobacterium-mediated transformation of durum wheat(Triticum turgidum L.var.durum cv Stewart)with improved efficiency,J.Exp.Bot.,61(6):1567-1581
Hensel G.,Marthe C.,and Kumlehn J.,2017,Agrobacterium-mediated transformation of wheat using immature embryos,Planta,1679(1):129-138
Jia H.N.,Du L.P.,Hu L.,Wang K.,Liu H.W.,and Ye X.G.,2017,Evaluation of plant regeneration ability of immature embryos from eight new wheat varieties or lines developed in Henan Province,Zhiwu Yichuan Ziyuan Xuebao(Journal of Plant Genetic Resources),18(4):728-733(贾华岚,杜丽璞,胡琳,王轲,刘宏伟,叶兴国,2017,河南省新育成8个小麦品种(系)幼胚培养再生性能评价,植物遗传资源学报,18(4):728-733)
Krenek P.,Samajova O.,Luptovciak I.,Doskocilova A.,Komis G.,and Samaj J.,2015,Transient plant transformation mediated by Agrobacterium tumefaciens:principles,methods and applications,Biotechnol.Adv.,33(6):1024-1042
Lei M.Y.,Xu W.G.,Li X.B.,Zhang Q.C.,Wang H.W.,Zhang L.,Fang Y.H.,Li Y.,and Li C.X.,2017,Effect of maize C4-specific photosynthesis genes on photosynthesis and drought resistance of Arabidopsis thaliana,Mailei Zuowu Xuebao(Journal of Triticeae Crops),37(1):108-115(雷明月,许为钢,李小博,张庆琛,王会伟,张磊,方宇辉,李艳,李春鑫,2017,玉米C4光合酶基因导入对拟南芥光合特性及抗旱性的影响,麦类作物学报,37(1):108-115)
Li M.H.,Chen W.,Xing L.P.,Xiao J.,Wang H.Y.,Cao A.Z.,and Wang X.E.,2010,Optimization of an Agrobacterium-mediated transformation system for wheat,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),8(2):388-392(李明浩,陈炜,邢莉萍,肖进,王海燕,曹爱忠,王秀娥,2010,农杆菌介导小麦遗传转化条件的优化,分子植物育种,8(2):388-392)
Li X.H.,Li L.,Song F.S.,Ni D.H.,Lu X.Z.,Li H.,Duan Y.B.,Wang X.F.,and Yang J.B.,2011,Condition optimization of Agrobacterium-mediated transgenic rice,Shengwuxue Zazhi(Journal of Biology),28(2):13-16(李笑寒,李莉,宋风顺,倪大虎,陆徐忠,李浩,段永波,汪秀峰,杨剑波,2011,根癌农杆菌介导的水稻转基因技术体系的优化,生物学杂志,28(2):13-16)
Ma J.H.,Zheng H.,Peng Y.,Liu Y.,and Chen J.,2011,A primary evaluation on Agrobacterium-mediated transformation efficiency of Chuannong wheat cultivars,Zhongzi(Seed),30(3):40-43(马建华,郑寒,彭姚,刘燕,陈静,2011,农杆菌介导的“川农”系列小麦品种遗传转化效率研究初探,种子,30(3):40-43)
Raziuddin,Bakht J.,Swati Z.A.,Shafi M.,Ullah F.,And F.U.,and Akmal M.,2010,Effect of cultivars and culture medium on callus formation and plant regeneration from mature embryos of wheat(Triticum aestivum L.),Pak.J.Bot.,42(1):639-652
Shi Z.Y.,Yin G.X.,Du L.P.,Tao L.L.,Xu H.J.,and Ye X.G.,2011,Plant regeneration and Agrobacterium-mediated transformation using large immature embryos of wheat,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),44(2):225-232(石珍源,殷桂香,杜丽璞,陶丽莉,徐惠君,叶兴国,2011,小麦大龄幼胚再生性能改进与农杆菌转化,中国农业科学,44(2):225-232)
Sparks C.A.,Doherty A.,and Jones H.D.,2014,Genetic transformation of wheat via Agrobacterium-mediated DNA delivery,Cereal Genomics,1099:235-250
Tang Y.,Wang Z.C.,Cui G.B.,Sun F.L.,Zhang C.,Liu S.D.,and Xi Y.J.,2016,Evaluation of regeneration and physiological characteristics of callus from mature embryo of wheat,Mailei Zuowu Xuebao(Journal of Triticeae Crops),36(10):1307-1314(汤益,王志成,崔桂宾,孙风丽,张超,刘曙东,奚亚军,2016,小麦成熟胚高效再生基因型筛选及愈伤组织生理特性评价,麦类作物学报,36(10):1307-1314)
Tao L.L.,Yin G.X.,Du L.P.,Shi Z.Y.,She M.Y.,Xu H.J.,and Ye X.G.,2011,Improvement of plant regeneration from immature embryos of wheat infected by Agrobacterium tumefaciens,Agricultural Sciences in China,10(3):317-326
Tester M.,and Langridge P.,2010,Breeding technologies to increase crop production in a changing world,Science,327(5967):818-822
Tian L.L.,Li J.,Zhang J.,Liu J.J.,Wang Y.Z.,Wang K.B.,Huang J.A.,and Liu Z.H.,2017,Studies on optimization of antibiotic species and concentration in genetic transformation of Camellia sinensis L.mediated by Agrobacterium,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),15(3):934-937(田丽丽,李娟,张静,刘建军,王英姿,王坤波,黄建安,刘仲华,2017,农杆菌介导茶树遗传转化中抗生素种类和浓度优化,分子植物育种,15(3):934-937)
Wang Y.Q.,Xiao X.G.,and Zhang A.M.,2002,Factors affecting Agrobacterium tumefaciens-mediated transformation of wheat(Triticum aestivum L.),Yichuan Xuebao(Acta Genetica Sinica),29(3):260-265(王永勤,肖兴国,张爱民,2002,农杆菌介导的小麦遗传转化几个影响因素的研究,遗传学报,29(3):260-265)
Wu B.H.,Zheng Y.L.,Zhou Y.H.,and Hu C.,2001,Variability of somatic embryogenesis capability of immature embryo calli from Sichuan wheat cultivars,Xinan Nongye Xuebao(Southwest China Journal of Agricultural Sciences),14(2):9-13(伍碧华,郑有良,周永红,胡超,2001,四川栽培小麦幼胚愈伤组织胚状体形成能力变异性研究,西南农业学报,14(2):9-13)
Wu H.N.,Zhang Y.S.,Wang X.L.,Zhao Y.H.,Wen S.M.,and Liu G.R.,2012,Optimization of wheat immature embryos genetic transformation system mediated by Agrobacterium tumefaciens,Mailei Zuowu Xuebao(Journal of Triticeae Crops),32(3):421-426(武海娜,张永升,王小龙,赵艳红,温树敏,刘桂茹,2012,农杆菌介导的小麦幼胚遗传转化体系的优化,麦类作物学报,32(3):421-426)
Xi Y.J.,Gao H.Z.,LüX.Y.,Lu M.,and Liu S.D.,2009,Genetic transformation system of wheat xiaoyan 22 mediated by Agrobacterium tumefaciens,Henongxue Bao(Journal of Nuclear Agricultural Sciences),23(2):185-192(奚亚军,高海战,吕晓依,路明,刘曙东,2009,黄淮麦区推广品种小偃22农杆菌遗传转化体系的建立,核农学报,23(2):185-192)
Ye X.G.,Wang X.M.,Wang K.,Du L.P.,Lin Z.S.,and Xu H.J.,2012,A review of some assisted strategies for improving the efficiency of Agrobacterium-mediated plant transformation,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),45(15):3007-3019(叶兴国,王新敏,王轲,杜丽璞,林志珊,徐惠君,2012,提高植物农杆菌转化效率辅助策略研究进展,中国农业科学,45(15):3007-3019)
Ye X.G.,Wang Y.L.,Kang L.,Du L.P.,and Xu H.J.,2005,Screening of wheat genotypes sensitive to Agrobacterium tumefaciens infection and transformation,Acta Agronomica Sinica,31(12):1552-1556
Ye X.G.,Xu H.J.,Du L.P.,He G.Y.,Wang K.,and Lin Z.S.,2014,Establishment and application of large-scale transformation systems in wheat,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),47(21):4155-4171(叶兴国,徐惠君,杜丽璞,何光源,王轲,林志珊,2014,小麦规模化转基因技术体系构建及其应用,中国农业科学,47(21):4155-4171)
Zhang C.,Wang M.,Li S.N.,Ma Y.L.,Wang L.S.,Jin Y.M.,Wang Z.,Li W.B.,Su A.Y.,and Wu X.X.,2015,Research progress on regeneration gene of ESR1 in plant,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),35(10):2270-2276(张超,王敏,李思楠,马彦龙,王玲爽,金杨媚,王智,李文滨,苏安玉,武小霞,2015,植物再生基因ESR1研究进展,基因组学与应用生物学,35(10):2270-2276)
Zhao P.,Wang K.,Zhang W.,Du L.P.,and Ye X.G.,2014,Review and inspiration of plant proteins involved in the transformation processing of T-DNA initiated by Agrobacterium,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),47(13):2504-2518(赵佩,王轲,张伟,杜丽璞,叶兴国,2014,参与农杆菌侵染及T-DNA转运过程植物蛋白的研究进展和思考,中国农业科学,47(13):2504-2518)
Zhou X.H.,Wang K.,Lv D.W.,Wu C.J.,Li J.R.,Zhao P.,Lin Z.S.,Du L.P.,Yan Y.M.,and Ye X.G.,2013,Global analysis of differentially expressed genes and proteins in the wheat callus infected by Agrobacterium tumefaciens,PLoS One,8(11):e79390
Zhu J.Y.,Xu Y.J.,Yang L.P.,Kang X.L.,Sun Y.,and Du J.Z.,2008,Study of wheat tissue culture regeneration system and genetic transformation mediated by Agrobacterium tumefaciens,Zhiwu Yichuan Ziyuan Xuebao(Journal of Plant Genetic Resources),9(1):84-89(朱晋云,许玉娟,杨丽萍,亢秀丽,孙毅,杜建忠,2008,小麦组织培养再生系统及农杆菌介导的转基因技术研究,植物遗传资源学报,9(1):84-89)
Ding L.,Li S.,Gao J.,Wang Y.,Yang G.,and He G.,2009,Optimization of Agrobacterium-mediated transformation conditions in mature embryos of elite wheat,Mol.Biol.Rep.,36(1):29-36
Du W.P.,Yu G.R.,Song J.,Xu L.Y.,Zhang L.,and Jiang Y.,2012,Effects of genotypes and explants on in vitro culture of wheat,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),10(3):290-297(杜文平,余桂容,宋军,徐利远,张莲,蒋云,2012,基因型和外植体对小麦离体培养的影响,分子植物育种,10(3):290-297)
Gao J.S.,Xu L.,Chen W.,Shen Z.L.,Cai Y.P.,and Lin Y.,2017,Agrobacterium-mediated genetic transformation of stem apexes of brown fiber cotton Zongcaixuan 1,Henongxue Bao(Journal of Nuclear Agricultural Sciences),31(10):1881-1888(高俊山,许磊,陈文,沈知临,蔡永萍,林毅,2017,农杆菌介导陆地棉棕彩选1号茎尖遗传转化,核农学报,31(10):1881-1888)
Godwin I.,Todd G.,Fordlloyd B.,and Newbury H.J.,1991,The effects of acetosyringone and p H on Agrobacterium-mediated transformation vary according to plant species,Plant Cell Rep.,9(12):671-675
He Y.,Jones H.D.,Chen S.,Chen X.M.,Wang D.W.,Li K.X.,Wang D.S.,and Xia L.Q.,2010,Agrobacterium-mediated transformation of durum wheat(Triticum turgidum L.var.durum cv Stewart)with improved efficiency,J.Exp.Bot.,61(6):1567-1581
Hensel G.,Marthe C.,and Kumlehn J.,2017,Agrobacterium-mediated transformation of wheat using immature embryos,Planta,1679(1):129-138
Jia H.N.,Du L.P.,Hu L.,Wang K.,Liu H.W.,and Ye X.G.,2017,Evaluation of plant regeneration ability of immature embryos from eight new wheat varieties or lines developed in Henan Province,Zhiwu Yichuan Ziyuan Xuebao(Journal of Plant Genetic Resources),18(4):728-733(贾华岚,杜丽璞,胡琳,王轲,刘宏伟,叶兴国,2017,河南省新育成8个小麦品种(系)幼胚培养再生性能评价,植物遗传资源学报,18(4):728-733)
Krenek P.,Samajova O.,Luptovciak I.,Doskocilova A.,Komis G.,and Samaj J.,2015,Transient plant transformation mediated by Agrobacterium tumefaciens:principles,methods and applications,Biotechnol.Adv.,33(6):1024-1042
Lei M.Y.,Xu W.G.,Li X.B.,Zhang Q.C.,Wang H.W.,Zhang L.,Fang Y.H.,Li Y.,and Li C.X.,2017,Effect of maize C4-specific photosynthesis genes on photosynthesis and drought resistance of Arabidopsis thaliana,Mailei Zuowu Xuebao(Journal of Triticeae Crops),37(1):108-115(雷明月,许为钢,李小博,张庆琛,王会伟,张磊,方宇辉,李艳,李春鑫,2017,玉米C4光合酶基因导入对拟南芥光合特性及抗旱性的影响,麦类作物学报,37(1):108-115)
Li M.H.,Chen W.,Xing L.P.,Xiao J.,Wang H.Y.,Cao A.Z.,and Wang X.E.,2010,Optimization of an Agrobacterium-mediated transformation system for wheat,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),8(2):388-392(李明浩,陈炜,邢莉萍,肖进,王海燕,曹爱忠,王秀娥,2010,农杆菌介导小麦遗传转化条件的优化,分子植物育种,8(2):388-392)
Li X.H.,Li L.,Song F.S.,Ni D.H.,Lu X.Z.,Li H.,Duan Y.B.,Wang X.F.,and Yang J.B.,2011,Condition optimization of Agrobacterium-mediated transgenic rice,Shengwuxue Zazhi(Journal of Biology),28(2):13-16(李笑寒,李莉,宋风顺,倪大虎,陆徐忠,李浩,段永波,汪秀峰,杨剑波,2011,根癌农杆菌介导的水稻转基因技术体系的优化,生物学杂志,28(2):13-16)
Ma J.H.,Zheng H.,Peng Y.,Liu Y.,and Chen J.,2011,A primary evaluation on Agrobacterium-mediated transformation efficiency of Chuannong wheat cultivars,Zhongzi(Seed),30(3):40-43(马建华,郑寒,彭姚,刘燕,陈静,2011,农杆菌介导的“川农”系列小麦品种遗传转化效率研究初探,种子,30(3):40-43)
Raziuddin,Bakht J.,Swati Z.A.,Shafi M.,Ullah F.,And F.U.,and Akmal M.,2010,Effect of cultivars and culture medium on callus formation and plant regeneration from mature embryos of wheat(Triticum aestivum L.),Pak.J.Bot.,42(1):639-652
Shi Z.Y.,Yin G.X.,Du L.P.,Tao L.L.,Xu H.J.,and Ye X.G.,2011,Plant regeneration and Agrobacterium-mediated transformation using large immature embryos of wheat,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),44(2):225-232(石珍源,殷桂香,杜丽璞,陶丽莉,徐惠君,叶兴国,2011,小麦大龄幼胚再生性能改进与农杆菌转化,中国农业科学,44(2):225-232)
Sparks C.A.,Doherty A.,and Jones H.D.,2014,Genetic transformation of wheat via Agrobacterium-mediated DNA delivery,Cereal Genomics,1099:235-250
Tang Y.,Wang Z.C.,Cui G.B.,Sun F.L.,Zhang C.,Liu S.D.,and Xi Y.J.,2016,Evaluation of regeneration and physiological characteristics of callus from mature embryo of wheat,Mailei Zuowu Xuebao(Journal of Triticeae Crops),36(10):1307-1314(汤益,王志成,崔桂宾,孙风丽,张超,刘曙东,奚亚军,2016,小麦成熟胚高效再生基因型筛选及愈伤组织生理特性评价,麦类作物学报,36(10):1307-1314)
Tao L.L.,Yin G.X.,Du L.P.,Shi Z.Y.,She M.Y.,Xu H.J.,and Ye X.G.,2011,Improvement of plant regeneration from immature embryos of wheat infected by Agrobacterium tumefaciens,Agricultural Sciences in China,10(3):317-326
Tester M.,and Langridge P.,2010,Breeding technologies to increase crop production in a changing world,Science,327(5967):818-822
Tian L.L.,Li J.,Zhang J.,Liu J.J.,Wang Y.Z.,Wang K.B.,Huang J.A.,and Liu Z.H.,2017,Studies on optimization of antibiotic species and concentration in genetic transformation of Camellia sinensis L.mediated by Agrobacterium,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),15(3):934-937(田丽丽,李娟,张静,刘建军,王英姿,王坤波,黄建安,刘仲华,2017,农杆菌介导茶树遗传转化中抗生素种类和浓度优化,分子植物育种,15(3):934-937)
Wang Y.Q.,Xiao X.G.,and Zhang A.M.,2002,Factors affecting Agrobacterium tumefaciens-mediated transformation of wheat(Triticum aestivum L.),Yichuan Xuebao(Acta Genetica Sinica),29(3):260-265(王永勤,肖兴国,张爱民,2002,农杆菌介导的小麦遗传转化几个影响因素的研究,遗传学报,29(3):260-265)
Wu B.H.,Zheng Y.L.,Zhou Y.H.,and Hu C.,2001,Variability of somatic embryogenesis capability of immature embryo calli from Sichuan wheat cultivars,Xinan Nongye Xuebao(Southwest China Journal of Agricultural Sciences),14(2):9-13(伍碧华,郑有良,周永红,胡超,2001,四川栽培小麦幼胚愈伤组织胚状体形成能力变异性研究,西南农业学报,14(2):9-13)
Wu H.N.,Zhang Y.S.,Wang X.L.,Zhao Y.H.,Wen S.M.,and Liu G.R.,2012,Optimization of wheat immature embryos genetic transformation system mediated by Agrobacterium tumefaciens,Mailei Zuowu Xuebao(Journal of Triticeae Crops),32(3):421-426(武海娜,张永升,王小龙,赵艳红,温树敏,刘桂茹,2012,农杆菌介导的小麦幼胚遗传转化体系的优化,麦类作物学报,32(3):421-426)
Xi Y.J.,Gao H.Z.,LüX.Y.,Lu M.,and Liu S.D.,2009,Genetic transformation system of wheat xiaoyan 22 mediated by Agrobacterium tumefaciens,Henongxue Bao(Journal of Nuclear Agricultural Sciences),23(2):185-192(奚亚军,高海战,吕晓依,路明,刘曙东,2009,黄淮麦区推广品种小偃22农杆菌遗传转化体系的建立,核农学报,23(2):185-192)
Ye X.G.,Wang X.M.,Wang K.,Du L.P.,Lin Z.S.,and Xu H.J.,2012,A review of some assisted strategies for improving the efficiency of Agrobacterium-mediated plant transformation,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),45(15):3007-3019(叶兴国,王新敏,王轲,杜丽璞,林志珊,徐惠君,2012,提高植物农杆菌转化效率辅助策略研究进展,中国农业科学,45(15):3007-3019)
Ye X.G.,Wang Y.L.,Kang L.,Du L.P.,and Xu H.J.,2005,Screening of wheat genotypes sensitive to Agrobacterium tumefaciens infection and transformation,Acta Agronomica Sinica,31(12):1552-1556
Ye X.G.,Xu H.J.,Du L.P.,He G.Y.,Wang K.,and Lin Z.S.,2014,Establishment and application of large-scale transformation systems in wheat,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),47(21):4155-4171(叶兴国,徐惠君,杜丽璞,何光源,王轲,林志珊,2014,小麦规模化转基因技术体系构建及其应用,中国农业科学,47(21):4155-4171)
Zhang C.,Wang M.,Li S.N.,Ma Y.L.,Wang L.S.,Jin Y.M.,Wang Z.,Li W.B.,Su A.Y.,and Wu X.X.,2015,Research progress on regeneration gene of ESR1 in plant,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),35(10):2270-2276(张超,王敏,李思楠,马彦龙,王玲爽,金杨媚,王智,李文滨,苏安玉,武小霞,2015,植物再生基因ESR1研究进展,基因组学与应用生物学,35(10):2270-2276)
Zhao P.,Wang K.,Zhang W.,Du L.P.,and Ye X.G.,2014,Review and inspiration of plant proteins involved in the transformation processing of T-DNA initiated by Agrobacterium,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),47(13):2504-2518(赵佩,王轲,张伟,杜丽璞,叶兴国,2014,参与农杆菌侵染及T-DNA转运过程植物蛋白的研究进展和思考,中国农业科学,47(13):2504-2518)
Zhou X.H.,Wang K.,Lv D.W.,Wu C.J.,Li J.R.,Zhao P.,Lin Z.S.,Du L.P.,Yan Y.M.,and Ye X.G.,2013,Global analysis of differentially expressed genes and proteins in the wheat callus infected by Agrobacterium tumefaciens,PLoS One,8(11):e79390
Zhu J.Y.,Xu Y.J.,Yang L.P.,Kang X.L.,Sun Y.,and Du J.Z.,2008,Study of wheat tissue culture regeneration system and genetic transformation mediated by Agrobacterium tumefaciens,Zhiwu Yichuan Ziyuan Xuebao(Journal of Plant Genetic Resources),9(1):84-89(朱晋云,许玉娟,杨丽萍,亢秀丽,孙毅,杜建忠,2008,小麦组织培养再生系统及农杆菌介导的转基因技术研究,植物遗传资源学报,9(1):84-89)