小黑麦花药培养效果研讨
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摘要
为完善小黑麦花药培养技术体系,本研究在项目组前期研究基础上,研究了低温(4℃)预处理时间(15 d, 20 d, 25 d, 30 d, 35 d, 40 d)和高温(32℃)培养时间(0 d, 1 d, 2 d, 3 d)对小黑麦花药培养的影响,以及小黑麦不同基因型(石大1号小黑麦品种,甘农1号小黑麦品种,小黑麦品系P4, TZ3, TZ10, TZ12, TZ23, TZ25, TZ26)的花药培养效果。结果显示:小黑麦幼穗低温4℃预处理25 d时,花药的出愈率最高达17.27%。花药高温32℃培养2 d时,石大1号出愈率最高(56%),而P4不进行预处理时最高;小黑麦基因型间花药出愈率及绿苗分化率有显著差异,TZ26、TZ25和P4的出愈率及绿苗分化率显著高于其余材料,石大1号的出愈率较高,但绿苗分化率低。本研究结果为小黑麦单倍体育种提供科学依据。
In order to improve the anther culture techniques of triticale, this research studied the effects of low temperature(4℃) pretreatment time(15 d, 20 d, 25 d, 30 d, 35 d, 40 d) and high temperature(32℃) culture time(0 d, 1 d, 2 d, 3 d) on the anther culture of triticale based on the previous researches of project team. Also, the anther culture effect of triticale with different genotypes(Shida No.1 triticale, Gannong No.1 triticale, triticale line P4, TZ3, TZ10, TZ12, TZ23, TZ25, TZ26) was analyzed. The results showed that the callus induction rate of anther of triticale young ear reached 17.27% under 4℃ pretreatment for 25 d. When being treated under 32℃ for 2 d,the callus induction rate of Shida No.1 triticale was the highest(56%), while that of P4 was the highest without pretreatment. The callus induction rate of anther and plantlet differentiation rate of triticale with different genotypes were quite different. The callus induction rate of anther and plantlet differentiation rate of TZ26, TZ25 and P4 were significantly higher than other materials, and Shida No.1 had higher callus induction rate but lower plantlet differentiation rate. The result of this study could provide scientific reference for the haploid breeding of triticale.
In order to improve the anther culture techniques of triticale, this research studied the effects of low temperature(4℃) pretreatment time(15 d, 20 d, 25 d, 30 d, 35 d, 40 d) and high temperature(32℃) culture time(0 d, 1 d, 2 d, 3 d) on the anther culture of triticale based on the previous researches of project team. Also, the anther culture effect of triticale with different genotypes(Shida No.1 triticale, Gannong No.1 triticale, triticale line P4, TZ3, TZ10, TZ12, TZ23, TZ25, TZ26) was analyzed. The results showed that the callus induction rate of anther of triticale young ear reached 17.27% under 4℃ pretreatment for 25 d. When being treated under 32℃ for 2 d,the callus induction rate of Shida No.1 triticale was the highest(56%), while that of P4 was the highest without pretreatment. The callus induction rate of anther and plantlet differentiation rate of triticale with different genotypes were quite different. The callus induction rate of anther and plantlet differentiation rate of TZ26, TZ25 and P4 were significantly higher than other materials, and Shida No.1 had higher callus induction rate but lower plantlet differentiation rate. The result of this study could provide scientific reference for the haploid breeding of triticale.
引文
Cao Y.,Li P.F.,Gan L.Y.,Sun N.,and Wei L.J.,2007,The effect of genotype and medium of anther culture of winterness triticale,Xinjiang Nongye Xuebao(Xinjiang Agricultural Science),44(S3):97-100(曹禹,李鹏飞,甘玲玉,孙娜,魏凌基,2007,冬性小黑麦花药培养的基因型和培养基效应的研究,新疆农业科学,44(S3):97-100)
Fu W.T.,Liao F.F.,He J.W.,Xing D.,Su D.,Huang D.F.,and Zhan Y.F.,2015,Effects of different temperatures and additives on anther culture of pepper,Nanfang Nongye Xuebao(Journal of Southern Agriculture),46(4):635-640(付文婷,廖芳芳,何建文,邢丹,苏丹,黄冬福,詹永发,2015,不同温度和外源添加物对辣椒花药的培养效果,南方农业学报,46(4):635-640)
GermanàM.A.,2011,Anther culture for haploid and doubled haploid production,Plant Cell Tiss.Org.Cult.,104(3):283-300
Guo X.M.,Ye K.C.,Pei Y.H.,and Song X.Y.,2013,Main affecting factors on cultural characteristics of maize anther,Yumi Kexue(Journal of Maize Sciences),21(4):83-88(郭新梅,叶克苁,裴玉贺,宋希云,2013,影响玉米花药培养特性关键因素的研究,玉米科学,21(4):83-88)
Han X.F.,Tao L.L.,Yin G.X.,Liu X.L.,Du L.P.,Wei Y.Q.,Yan Y.M.,and Ye X.G.,2010,Effect of genotype and growing environment on anther culture in wheat,Zuowu Xuebao(Acta Agronomica Sinica),36(7):1209-1215(韩晓峰,陶丽莉殷桂香,刘晓蕾,杜丽璞,魏亦勤,晏月明,叶兴国,2010基因型和环境条件对小麦花药培养效果的影响,作物学报,36(7):1209-1215)
Hua W.,Shang Y.,Jia Q.J.,Wang J.M.,Zhu J.H.,Lin F.,Gu Y.K.and Yang J.M.,2013,Rapid culture method for barley anther and effect of genotype difference in anther culture response,Zhejiang Nongye Xuebao(Acta Agriculturae Zhejiangensis),25(3):425-430(华为,尚毅,贾巧君,汪军妹,朱靖环,林峰,顾玉坤,杨建明,2013,大麦花药快速培养法及不同基因型对花药培养的影响,浙江农业学报,25(3)425-430)
Kikuchi K.,Terauchi K.,Wada M.,and Hirano H.Y.,2003,The plant MITE m Ping is mobilized in anther culture,Nature,421(6919):167-170
Lv X.L.,Bai H.B.,Dong J.L.,Cai Z.Y.,He H.Y.,and Li S.H.2015,Effects of medium and the darkness culture time of high-temperature on wheat anther culture,Zhongzi(Seed)34(6):84-87(吕学莲,白海波,董建力,蔡正云,何海洋,李树华,2015,培养基及高温暗培养时间对小麦花药培养的影响,种子,34(6):84-87)
Ma J.L.,and Zhang B.,2007,Effects of cold pretreatment and low temperature pre-culture on the callus induction rate on anther culture of Alfalfa,Zhongguo Caodi Xuebao(Chinese Journal of Grassland),29(6):59-63(马菊兰,张博,2007,低温预处理和低温预培养对苜蓿新品系花药愈伤组织诱导的影响,中国草地学报,29(6):59-63)
Marciniak K.,Kaczmarek Z.,Adamski T.,and Surma M.,2003The anther-culture response of triticale line x tester progenies,Cell.Mol.Biol.Lett.,8(2):343
Ni S.L.,Li W.,Li S.,Chai S.X.,and Li Z.M.,2004,Factors influencing on high efficiency anthers culture of wheat in dryland,Gansu Nongye Daxue Xuebao(Journal of Gansu Agricultural University),39(2):141-145(倪胜利,李唯,李胜柴守玺,李志明,2004,影响旱作小麦高效花药培养的因素,甘肃农业大学学报,39(2):141-145)
Song Q.,Tian X.H.,and Du W.H.,2016,Studies on production performance of new triticale lines in alpine pastoral areas of Gansu,Caoye Kexue(Pratacultural Sciencce),33(7):1367-1374(宋谦,田新会,杜文华,2016,甘肃省高寒牧区小黑麦新品系的生产性能,草业科学,33(7):1367-1374)
Song Y.X.,Zhou S.Y.,Du X.L.,Wang J.,Yu R.G.,Li G.P.Wang Q.,and Chen Y.F.,2012,Effect of low temperature pretreatment on the variation of endogenous hormones in wheat anther,Henongxue Bao(Journal of Nuclear Agricultural Sciences),26(7):1064-1069(宋运贤,周素英,杜雪玲,王建,余如刚,李桂萍,王倩,陈耀锋,2012,低温预处理对小麦花药内源激素的影响,核农学报,26(7):1064-1069)
Sun N.,Shi P.J.,Wei L.J.,Cao Y.,Huang R.,Li S.M.,and Liang W.,2009a,Genetic control analysis of triticale anther culture response,Mailei Zuowu Xuebao(Journal of Triticeae Crops),29(3):374-379(孙娜,石培春,魏凌基,曹禹,黄润,李守明,梁唯,2009a,小黑麦花药培养力的遗传模式分析,麦类作物学报,29(3):374-379)
Sun N.,Shi P J.,Wei L.J.,Cao Y.,Gan L.Y.,and Li P.F.,2009b,Study on factors affecting the induction of anther callus in winter and spring triticale,Shihezi Daxue Xuebao(Journal of Shihezi University(Natural Science)),27(1):1-5(孙娜,石培春,魏凌基,曹禹,甘铃玉,李鹏飞,2009b,冬春性小黑麦花药愈伤组织诱导的影响因素,石河子大学学报(自然科学版),27(1):1-5)
Torp A.M.,Hansen A.L.,and Andersen S.B.,2001,Chromosomal regions associated with green plant regeneration in wheat(Triticum aestivum L.)anther culture,Euphytica,119(3):377-387,
Wang W.,Chen C.,Ou Q.M.,Ye C.L.,and Luo J.J.,2016,Research and application of wheat anther culture,Henongxue Bao(Journal of Nuclear Agricultural Sciences),30(12):2343-235(王炜,陈琛,欧巧明,叶春雷,罗俊杰,2016,小麦花药培养的研究和应用,核农学报,30(12):2343-2354)
Wang Y.,Fan Q.Q.,Zhang L.,Sui X.X.,Li G.Y.,Chu X.S.,Zhang X.S.,and Huang C.Y.,2007,Quantitative genetic analysis of anther culture ability in wheat,Mailei Zuowu Xuebao(Journal of Triticeae Crops),27(5):755-760(王羽,樊庆琦,张利,隋新霞,李根英,楚秀生,张宪省,黄承彦,2007,小麦花药培养特性的数量遗传分析,麦类作物学报,27(5):755-760)
Yang X.,Liu C.L.,Wang D.,Zhang L.Q.,Wang S.J.,Chang H.P.,Xing W.H.,and Fu R.M.,2015,Effects of low temperature pretreatment and PEG on callus induction rate of wheat anther,Henan Nongye Xuebao(Journal of Henan Agricultural Sciences),44(4):49-51(杨雪,刘春雷,王丁,张丽琴,王世杰,常慧萍,邢文会,付瑞敏,2015,低温预处理和聚乙二醇处理对小麦花药出愈率的影响,河南农业科学,44(4):49-51)
Yao J.L.,Guo B.,and Xie H.,2015,Expression analysis of key enzyme gene involved in Chlorophyll biosynthesis by semi-quantitative RT-PCR,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(3):593-598(姚嘉龙,郭蓓,谢皓,2015,叶绿素合成关键酶基因表达的半定量RT-PCR分析,基因组学与应用生物学,34(3):593-598)
Zhang F.Y.,2017,Evaluations on drought resistance and studies on the production of new triticale lines in arid and semi-arid irrigated area,Thesis for M.S.,Gansu Agricultural University,Supervisor:Du W.H.,pp.1-25(赵方媛,2017,饲料型小黑麦新品系的抗旱性评价及在干旱半干旱灌区的生产性能研究,硕士学位论文,甘肃农业大学,导师:杜文华,pp.1-25)
Zhang G.L.,Zhang S.T.,Xiao L.T.,Wu X.J.,Xiao Y.H.,and Chen L.Y.,2013,Effect of high temperature stress on physiological characteristics of anther and pollen traits of rice a flowering stage,Zuowu Xuebao(Acta Agronomica Sinica)39(1):177-183(张桂莲,张顺堂,肖浪涛,武小金,肖应辉陈立云,2013,花期高温胁迫对水稻花药生理特性及花粉性状的影响,作物学报,39(1):177-183)
Zhang G.L.,Chen L.Y.,Zhang S.T.,Liu G.H.,Tang W.B.,Li MH.,Lei D.Y.,and Chen X.B.,2008,Effects of high temperature stress on pollen characters and anther microstructure of rice,Shengtai Xuebao(Acta Ecologica Sinica),28(3):1089-1097(张桂莲,陈立云,张顺堂,刘国华,唐文邦,李梅华,雷东阳,陈信波,2008,高温胁迫对水稻花粉粒性状及花药显微结构的影响,生态学报,28(3):1089-1097)
Zhao L.S.,Liu L.X.,Guo H.J.,Gu J.Y.,Zhao S.R.,Li J.H.,and Xie Y.D.,2012,Combining ability analysis on anther culture traits of three wheat genotypes with high regeneration ability,Mailei Zuowu Xuebao(Journal of Triticeae Crops),32(3):427-430(赵林姝,刘录祥,郭会君,古佳玉,赵世荣,李军辉,谢永盾,2012,三个高花药培养力小麦材料培养力性状的配合力分析,麦类作物学报,32(3):427-430)
Zhuang J.P.,Gong Z.H.,and Su J.,2001,The effects of temperature treatment on callus induction of Capsicum Anuum L.,Xibei Nongye Xuebao(Acta Agriculturae Boreali-Occidentalis Sinica),10(2):49-51(庄军平,巩振辉,苏菁,2001,温度对辣椒花药愈伤组织形成的影响,西北农业学报,10(2):49-51)
Fu W.T.,Liao F.F.,He J.W.,Xing D.,Su D.,Huang D.F.,and Zhan Y.F.,2015,Effects of different temperatures and additives on anther culture of pepper,Nanfang Nongye Xuebao(Journal of Southern Agriculture),46(4):635-640(付文婷,廖芳芳,何建文,邢丹,苏丹,黄冬福,詹永发,2015,不同温度和外源添加物对辣椒花药的培养效果,南方农业学报,46(4):635-640)
GermanàM.A.,2011,Anther culture for haploid and doubled haploid production,Plant Cell Tiss.Org.Cult.,104(3):283-300
Guo X.M.,Ye K.C.,Pei Y.H.,and Song X.Y.,2013,Main affecting factors on cultural characteristics of maize anther,Yumi Kexue(Journal of Maize Sciences),21(4):83-88(郭新梅,叶克苁,裴玉贺,宋希云,2013,影响玉米花药培养特性关键因素的研究,玉米科学,21(4):83-88)
Han X.F.,Tao L.L.,Yin G.X.,Liu X.L.,Du L.P.,Wei Y.Q.,Yan Y.M.,and Ye X.G.,2010,Effect of genotype and growing environment on anther culture in wheat,Zuowu Xuebao(Acta Agronomica Sinica),36(7):1209-1215(韩晓峰,陶丽莉殷桂香,刘晓蕾,杜丽璞,魏亦勤,晏月明,叶兴国,2010基因型和环境条件对小麦花药培养效果的影响,作物学报,36(7):1209-1215)
Hua W.,Shang Y.,Jia Q.J.,Wang J.M.,Zhu J.H.,Lin F.,Gu Y.K.and Yang J.M.,2013,Rapid culture method for barley anther and effect of genotype difference in anther culture response,Zhejiang Nongye Xuebao(Acta Agriculturae Zhejiangensis),25(3):425-430(华为,尚毅,贾巧君,汪军妹,朱靖环,林峰,顾玉坤,杨建明,2013,大麦花药快速培养法及不同基因型对花药培养的影响,浙江农业学报,25(3)425-430)
Kikuchi K.,Terauchi K.,Wada M.,and Hirano H.Y.,2003,The plant MITE m Ping is mobilized in anther culture,Nature,421(6919):167-170
Lv X.L.,Bai H.B.,Dong J.L.,Cai Z.Y.,He H.Y.,and Li S.H.2015,Effects of medium and the darkness culture time of high-temperature on wheat anther culture,Zhongzi(Seed)34(6):84-87(吕学莲,白海波,董建力,蔡正云,何海洋,李树华,2015,培养基及高温暗培养时间对小麦花药培养的影响,种子,34(6):84-87)
Ma J.L.,and Zhang B.,2007,Effects of cold pretreatment and low temperature pre-culture on the callus induction rate on anther culture of Alfalfa,Zhongguo Caodi Xuebao(Chinese Journal of Grassland),29(6):59-63(马菊兰,张博,2007,低温预处理和低温预培养对苜蓿新品系花药愈伤组织诱导的影响,中国草地学报,29(6):59-63)
Marciniak K.,Kaczmarek Z.,Adamski T.,and Surma M.,2003The anther-culture response of triticale line x tester progenies,Cell.Mol.Biol.Lett.,8(2):343
Ni S.L.,Li W.,Li S.,Chai S.X.,and Li Z.M.,2004,Factors influencing on high efficiency anthers culture of wheat in dryland,Gansu Nongye Daxue Xuebao(Journal of Gansu Agricultural University),39(2):141-145(倪胜利,李唯,李胜柴守玺,李志明,2004,影响旱作小麦高效花药培养的因素,甘肃农业大学学报,39(2):141-145)
Song Q.,Tian X.H.,and Du W.H.,2016,Studies on production performance of new triticale lines in alpine pastoral areas of Gansu,Caoye Kexue(Pratacultural Sciencce),33(7):1367-1374(宋谦,田新会,杜文华,2016,甘肃省高寒牧区小黑麦新品系的生产性能,草业科学,33(7):1367-1374)
Song Y.X.,Zhou S.Y.,Du X.L.,Wang J.,Yu R.G.,Li G.P.Wang Q.,and Chen Y.F.,2012,Effect of low temperature pretreatment on the variation of endogenous hormones in wheat anther,Henongxue Bao(Journal of Nuclear Agricultural Sciences),26(7):1064-1069(宋运贤,周素英,杜雪玲,王建,余如刚,李桂萍,王倩,陈耀锋,2012,低温预处理对小麦花药内源激素的影响,核农学报,26(7):1064-1069)
Sun N.,Shi P.J.,Wei L.J.,Cao Y.,Huang R.,Li S.M.,and Liang W.,2009a,Genetic control analysis of triticale anther culture response,Mailei Zuowu Xuebao(Journal of Triticeae Crops),29(3):374-379(孙娜,石培春,魏凌基,曹禹,黄润,李守明,梁唯,2009a,小黑麦花药培养力的遗传模式分析,麦类作物学报,29(3):374-379)
Sun N.,Shi P J.,Wei L.J.,Cao Y.,Gan L.Y.,and Li P.F.,2009b,Study on factors affecting the induction of anther callus in winter and spring triticale,Shihezi Daxue Xuebao(Journal of Shihezi University(Natural Science)),27(1):1-5(孙娜,石培春,魏凌基,曹禹,甘铃玉,李鹏飞,2009b,冬春性小黑麦花药愈伤组织诱导的影响因素,石河子大学学报(自然科学版),27(1):1-5)
Torp A.M.,Hansen A.L.,and Andersen S.B.,2001,Chromosomal regions associated with green plant regeneration in wheat(Triticum aestivum L.)anther culture,Euphytica,119(3):377-387,
Wang W.,Chen C.,Ou Q.M.,Ye C.L.,and Luo J.J.,2016,Research and application of wheat anther culture,Henongxue Bao(Journal of Nuclear Agricultural Sciences),30(12):2343-235(王炜,陈琛,欧巧明,叶春雷,罗俊杰,2016,小麦花药培养的研究和应用,核农学报,30(12):2343-2354)
Wang Y.,Fan Q.Q.,Zhang L.,Sui X.X.,Li G.Y.,Chu X.S.,Zhang X.S.,and Huang C.Y.,2007,Quantitative genetic analysis of anther culture ability in wheat,Mailei Zuowu Xuebao(Journal of Triticeae Crops),27(5):755-760(王羽,樊庆琦,张利,隋新霞,李根英,楚秀生,张宪省,黄承彦,2007,小麦花药培养特性的数量遗传分析,麦类作物学报,27(5):755-760)
Yang X.,Liu C.L.,Wang D.,Zhang L.Q.,Wang S.J.,Chang H.P.,Xing W.H.,and Fu R.M.,2015,Effects of low temperature pretreatment and PEG on callus induction rate of wheat anther,Henan Nongye Xuebao(Journal of Henan Agricultural Sciences),44(4):49-51(杨雪,刘春雷,王丁,张丽琴,王世杰,常慧萍,邢文会,付瑞敏,2015,低温预处理和聚乙二醇处理对小麦花药出愈率的影响,河南农业科学,44(4):49-51)
Yao J.L.,Guo B.,and Xie H.,2015,Expression analysis of key enzyme gene involved in Chlorophyll biosynthesis by semi-quantitative RT-PCR,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(3):593-598(姚嘉龙,郭蓓,谢皓,2015,叶绿素合成关键酶基因表达的半定量RT-PCR分析,基因组学与应用生物学,34(3):593-598)
Zhang F.Y.,2017,Evaluations on drought resistance and studies on the production of new triticale lines in arid and semi-arid irrigated area,Thesis for M.S.,Gansu Agricultural University,Supervisor:Du W.H.,pp.1-25(赵方媛,2017,饲料型小黑麦新品系的抗旱性评价及在干旱半干旱灌区的生产性能研究,硕士学位论文,甘肃农业大学,导师:杜文华,pp.1-25)
Zhang G.L.,Zhang S.T.,Xiao L.T.,Wu X.J.,Xiao Y.H.,and Chen L.Y.,2013,Effect of high temperature stress on physiological characteristics of anther and pollen traits of rice a flowering stage,Zuowu Xuebao(Acta Agronomica Sinica)39(1):177-183(张桂莲,张顺堂,肖浪涛,武小金,肖应辉陈立云,2013,花期高温胁迫对水稻花药生理特性及花粉性状的影响,作物学报,39(1):177-183)
Zhang G.L.,Chen L.Y.,Zhang S.T.,Liu G.H.,Tang W.B.,Li MH.,Lei D.Y.,and Chen X.B.,2008,Effects of high temperature stress on pollen characters and anther microstructure of rice,Shengtai Xuebao(Acta Ecologica Sinica),28(3):1089-1097(张桂莲,陈立云,张顺堂,刘国华,唐文邦,李梅华,雷东阳,陈信波,2008,高温胁迫对水稻花粉粒性状及花药显微结构的影响,生态学报,28(3):1089-1097)
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