箭竹DNA导入水稻后其遗传背景、农艺性状及产量分析
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摘要
【目的】将箭竹DNA导入水稻,并分析其遗传背景、农艺性状及产量,为水稻遗传育种提供技术参考。【方法】利用花粉管通道技术将箭竹DNA导入4个水稻恢复系受体材料(9311、华占、Q7和扬恢22)中,筛选出表型与受体材料存在差异的株系连续自交直至稳定,并利用筛选出的49对多态性SSR引物鉴定变异株系的遗传背景差异。以不同遗传背景的变异株系与不育系3S和6303S配制杂交组合,并比较分析其农艺性状差异。【结果】将箭竹DNA导入水稻后通过连续自交筛选出能稳定遗传的变异株系21个(T1~T21),其中9311、华占、Q7和扬恢22的变异率分别为4.85%、1.30%、2.35%和2.65%。利用SSR标记分析变异株系的遗传背景,发现其与受体材料在49对SSR标记在中存在差异的引物对数为2~24对,其中在24对标准引物中存在差异的对数范围为0~11对,表明通过花粉管通道技术产生的变异具有一定的随机性。不同遗传背景变异株系与不育系组配的杂交组合在农艺性状和产量方面存在明显差异,从中筛选出2个优势组合6303S×T10和6303S×T20,分别比我国长江中下游中籼迟熟组筛选试验对照品种丰两优4号增产10.9%和12.9%。【结论】将箭竹DNA导入水稻可产生稳定遗传的变异株系,从而获得与其组配的优势杂交组合,即通过花粉管通道技术导入外源DNA或基因是水稻种质资源创新及新品种选育的有效途径。
【Objective】DNA of Fargesia spathacea Franch. were introgressed into rice by pollen-tube pathway,then genetic background,agronomic traits and yield of rice were analyzed to provide reference for rice genetics and breeding.【Method】DNA of F. spathacea were introgressed into four rice varieties(9311,Huazhan,Q7 and Yanghui 22)by pollentube pathway. The lines whose phenotypes were different from receptor materials were selected to self-breed till they were stable. Then genetic background differences between variant lines and receptor materials were identified by the screened 49 pairs of SSR markers. Cross combinations were established by variant lines with different genetic background and two sterile lines(3S and 6303S),and their differences in agronomic traits were analyzed.【Result】The results showed that after DNA of F. spathacea were introgressed into rice receptor materials and let them self-bred,21 variant lines(T1-T21)that could transmit stably were obtained. Among them,variations of 9311,Huazhan,Q7 and Yanghui 22 were 4.85%,1.30%,2.35% and 2.65% respectively. Genetic background of the variant lines were analyzed by SSR markers. Among the 49 pairs of SSR markers,the variant lines were different from the receptor materials in 2-24 pairs of SSR markers;and among 24 pairs of standard primers,there were differences in 0-11 pairs. The results showed variation brought by pollen-tube pathway was random. The hybrid combinations by variant lines with various genetic backgrounds and sterile lines had various agronomic traits and yields. Two advantage hybrid combinations,6303S×T10 and 6303S×T20,from the cross combinations were selected. The yields of 6303S×T10 and 6303S×T20 increased by 10.9% and 12.9% than Fengliangyou 4,a control variety selected from middle-season late-mature indica rice combinations in the middle and lower reaches of Yangtze River.【Conclusion】Introgression of DNA of F. spathacea into rice can obtain variant lines with stable heredity,then obtain advantage hybrid combinations that crosses with them. Introgression of exogenous gene by pollen-tube pathway is an effective way in rice gerplasm resources innovation and variety breeding.
【Objective】DNA of Fargesia spathacea Franch. were introgressed into rice by pollen-tube pathway,then genetic background,agronomic traits and yield of rice were analyzed to provide reference for rice genetics and breeding.【Method】DNA of F. spathacea were introgressed into four rice varieties(9311,Huazhan,Q7 and Yanghui 22)by pollentube pathway. The lines whose phenotypes were different from receptor materials were selected to self-breed till they were stable. Then genetic background differences between variant lines and receptor materials were identified by the screened 49 pairs of SSR markers. Cross combinations were established by variant lines with different genetic background and two sterile lines(3S and 6303S),and their differences in agronomic traits were analyzed.【Result】The results showed that after DNA of F. spathacea were introgressed into rice receptor materials and let them self-bred,21 variant lines(T1-T21)that could transmit stably were obtained. Among them,variations of 9311,Huazhan,Q7 and Yanghui 22 were 4.85%,1.30%,2.35% and 2.65% respectively. Genetic background of the variant lines were analyzed by SSR markers. Among the 49 pairs of SSR markers,the variant lines were different from the receptor materials in 2-24 pairs of SSR markers;and among 24 pairs of standard primers,there were differences in 0-11 pairs. The results showed variation brought by pollen-tube pathway was random. The hybrid combinations by variant lines with various genetic backgrounds and sterile lines had various agronomic traits and yields. Two advantage hybrid combinations,6303S×T10 and 6303S×T20,from the cross combinations were selected. The yields of 6303S×T10 and 6303S×T20 increased by 10.9% and 12.9% than Fengliangyou 4,a control variety selected from middle-season late-mature indica rice combinations in the middle and lower reaches of Yangtze River.【Conclusion】Introgression of DNA of F. spathacea into rice can obtain variant lines with stable heredity,then obtain advantage hybrid combinations that crosses with them. Introgression of exogenous gene by pollen-tube pathway is an effective way in rice gerplasm resources innovation and variety breeding.
引文
陈洪伟.2008.白杨花粉管通道导入外源胡杨DNA技术研究[D].北京:北京林业大学.[Chen H W.2008.Introduction of exogenous DNA of Populus euphrtica into white poplars via pollen-tube pathway[D].Beijing:Beijing Forestry University.]
丁建庭,申家恒,李伟,杨虹.2009.水稻双受精过程的细胞形态学及时间进程的观察[J].植物学报,44(4):473-483.[Ding J T,Shen J H,Li W,Yang H.2009.Cytological observation of double fertilization and its duration in Oryza sativa[J].Chinese Bulletin of Botany,44(4):473-483.]
董春林,张明义,林忠平,张彦芹,杨丽莉,梁改梅,孙洁.2011.用花粉管通道法将抗旱耐盐基因导入玉米自交系的研究[J].山西农业科学,39(5):392-394.[Dong C L,Zhang M Y,Lin Z P,Zhang Y Q,Yang L L,Liang G M,Sun J.2011.Study on transferring gene of salt tolerance and drought resistance into corn inbred line by pollen tube path[J].Journal of Shanxi Agricultural Sciences,39(5):392-394.]
高树仁,王霞,孙丽芳,钱海霞,赵伟,刘芳.2016.玉米导入碱蓬DNA后代主要农艺性状和品质性状的变异分析[J].黑龙江八一农垦大学学报,28(5):9-13.[Gao S R,Wang X,Sun L F,Qian H X,Zhao W,Liu F.2016.Analysis of variation on main agronomic traits and quality traits of offspring of leading Suaeda DNA into maize[J].Journal of Heilongjiang Bayi Agricultural University,28(5):9-13.]
贾影影,刘凤楼,王掌军,刘生祥,陈丽萍,郭丽静.2016.大豆DNA导入春小麦后的农艺性状和品质性状分析及RAPD标记验证[J].麦类作物学报,36(7):833-840.[Jia Y Y,Liu F L,Wang Z J,Liu S X,Chen L P,Guo L J.2016.Study on agronomic and quality traits and RAPD marker analysis of transgenic spring wheat with soybean DNA[J].Journal of Triticeae Crops,36(7):833-840.]
刘冬梅,武芝霞,刘传亮,李成伟,李付广.2007.花粉管通道法获得棉花转基因株系主要农艺性状变异分析[J].棉花学报,19(6):450-454.[Liu D M,Wu Z X,Liu C L,Li C W,Li F G.2007.Variations of main agronomic traits in transgenic cotton(Gossypium hirsutum L.)lines transformed by pollen-tube pathway method[J].Cotton Science,19(6):450-454.]
倪建福,令利军,欧巧明,叶春雷,武禄光.2005.外源DNA导入小麦的分子育种实践[J].麦类作物学报,25(5):27-31.[Ni J F,Ling L J,Ou Q M,Ye C L,Wu L G.2005.Molecular breeding by introducing exogenous DNA into wheat[J].Journal of Triticeae Crops,25(5):27-31.]
孙一丹,胡远艺,彭彦,毛毕刚,王津津,赵炳然.2013.水稻外源DNA导入系变异系与受体的农艺性状差异及基因变异分析[J].杂交水稻,28(2):62-67.[Sun Y D,Hu Y Y,Peng Y,Mao B G,Wang J J,Zhao B R.2013.Phenotypic difference and gene variation in agronomic traits between a rice variant with introduced exogenous genomic DNA and its receptor[J].Hybrid Rice,28(2):62-67.]
唐清杰,云勇,张吉贞,严小微,林尤珍,徐靖.2010.海南疣粒野生稻DNA导入栽培稻及其后代的SSR验证[J].亚热带植物科学,39(4):7-9.[Tang Q J,Yun Y,Zhang J J,Yan X W,Lin Y Z,Xu J.2010.Rice transformation by injecting Hainan Oryza meyeriana DNA and SSR detection[J].Subtropical Plant Science,39(4):7-9.]
王丰,柳武革,李金华,刘振荣,李曙光,廖亦龙,彭惠香,毛兴学.2004.水稻外源DNA导入系的创建及主要农艺性状分析[J].分子植物育种,2(4):489-494.[Wang F,Liu W G,Li J H,Liu Z R,Li S G,Liao Y L,Peng H X,Mao X X.2004.Construction of exogenous DNA introgression lines and analysis of its main traits in rice[J].Molecular Plant Breeding,2(4):489-494.]
王永斌,赵远玲,闫丽娟,猛庆用,王广金.2011.玉米DNA导入水稻变异后代的研究[J].中国农学通报,27(7):63-68.[Wang Y B,Zhao Y L,Yan L J,Meng Q Y,Wang GJ.2011.Study on variation offspring from different rice varieties introduced DNA of maize[J].Chinese Agricultural Science Bulletin,27(7):63-68.]
魏霞,李云峰,向佳,凌英华,杨正林,何光华,赵芳明.2015.干旱胁迫对导入高粱DNA的水稻变异系R21生理的影响[J].西南农业学报,28(3):930-934.[Wei X,Li Y F,Xiang J,Ling Y H,Yang Z L,He G H,Zhao F M.2015.Effect of drought stress on physiological characters of rice variant line R21 by introgression sorghum DNA[J].Southwest China Journal of Agricultural Sciences,28(3):930-934.]
赵鑫闻.2016.利用花粉管通道技术将外源银白杨DNA导入黑杨[J].植物学报,51(4):533-541.[Zhao X W.2016.Introduction of exogenous DNA of Populus alba into Sect.Aigeiros via pollen-tube pathway[J].Plant Biology,51(4):533-541.]
肇莹,肖军,杨镇,杨涛.2015.转导芦苇总DNA耐盐变异水稻SSH文库的构建及部分基因特异表达[J].江苏农业科学,43(3):13-16.[Zhao Y,Xiao J,Yang Z,Yang T.2015.Construction of SSH libraries and partial gene specific expression in variant rice with salt tolerance transduced from total DNA of reed[J].Jiangsu Agricultural Sciences,43(3):13-16.]
周光宇,翁坚,龚蓁蓁,曾以申,杨晚霞,沈慰芳,王自芬,陶全洲,黄骏麒,钱思颖,刘桂玲,应苗成,薛达元,洪爱华,徐英俊,陈善葆,段晓岚.1988.农业分子育种授粉后外源DNA导入植物的技术[J].中国农业科学,21(3):1-6.[Zhou G Y,Weng J,Gong Z Z,Zeng Y S,Yang W X,Shen W F,Wang Z F,Tao Q Z,Huang J Q,Qian S Y,Liu G L,Ying M C,Xue D Y,Hong A H,Xu Y J,Chen S B,Duan X L.1988.Molecular breeding of agriculture a technique for introducing exogenous DNA into plants after self pollination[J].Scientia Agricultura Sinica,21(3):1-6.]
周岩,游建,魏琦超,薛晓锋,何男男,马俊.2011.花粉管通道法转基因对小麦主要农艺性状的影响[J].河南科技学院学报,39(4):5-8.[Zhou Y,You J,Wei Q C,Xue X F,He N N,Ma J.2011.Study on the influence of agronomic traits of transformed wheat by pollen tube pathway[J].Journal of Henan Institute of Science and Technology,39(4):5-8.]
Luo Z X,Wu R.1989.A simple method for the transformation of rice via the pollen-tube pathway[J].Plant Moecular Bio1ogy Reporter,7(3):240.
Zhou G Y,Weng J,Zeng Y S,Huang J G,Qiao S Y,Liu GL.1983.Introduction of exogenous DNA into cotton embryos[J].Methods in Enzymology,101:433-481.
丁建庭,申家恒,李伟,杨虹.2009.水稻双受精过程的细胞形态学及时间进程的观察[J].植物学报,44(4):473-483.[Ding J T,Shen J H,Li W,Yang H.2009.Cytological observation of double fertilization and its duration in Oryza sativa[J].Chinese Bulletin of Botany,44(4):473-483.]
董春林,张明义,林忠平,张彦芹,杨丽莉,梁改梅,孙洁.2011.用花粉管通道法将抗旱耐盐基因导入玉米自交系的研究[J].山西农业科学,39(5):392-394.[Dong C L,Zhang M Y,Lin Z P,Zhang Y Q,Yang L L,Liang G M,Sun J.2011.Study on transferring gene of salt tolerance and drought resistance into corn inbred line by pollen tube path[J].Journal of Shanxi Agricultural Sciences,39(5):392-394.]
高树仁,王霞,孙丽芳,钱海霞,赵伟,刘芳.2016.玉米导入碱蓬DNA后代主要农艺性状和品质性状的变异分析[J].黑龙江八一农垦大学学报,28(5):9-13.[Gao S R,Wang X,Sun L F,Qian H X,Zhao W,Liu F.2016.Analysis of variation on main agronomic traits and quality traits of offspring of leading Suaeda DNA into maize[J].Journal of Heilongjiang Bayi Agricultural University,28(5):9-13.]
贾影影,刘凤楼,王掌军,刘生祥,陈丽萍,郭丽静.2016.大豆DNA导入春小麦后的农艺性状和品质性状分析及RAPD标记验证[J].麦类作物学报,36(7):833-840.[Jia Y Y,Liu F L,Wang Z J,Liu S X,Chen L P,Guo L J.2016.Study on agronomic and quality traits and RAPD marker analysis of transgenic spring wheat with soybean DNA[J].Journal of Triticeae Crops,36(7):833-840.]
刘冬梅,武芝霞,刘传亮,李成伟,李付广.2007.花粉管通道法获得棉花转基因株系主要农艺性状变异分析[J].棉花学报,19(6):450-454.[Liu D M,Wu Z X,Liu C L,Li C W,Li F G.2007.Variations of main agronomic traits in transgenic cotton(Gossypium hirsutum L.)lines transformed by pollen-tube pathway method[J].Cotton Science,19(6):450-454.]
倪建福,令利军,欧巧明,叶春雷,武禄光.2005.外源DNA导入小麦的分子育种实践[J].麦类作物学报,25(5):27-31.[Ni J F,Ling L J,Ou Q M,Ye C L,Wu L G.2005.Molecular breeding by introducing exogenous DNA into wheat[J].Journal of Triticeae Crops,25(5):27-31.]
孙一丹,胡远艺,彭彦,毛毕刚,王津津,赵炳然.2013.水稻外源DNA导入系变异系与受体的农艺性状差异及基因变异分析[J].杂交水稻,28(2):62-67.[Sun Y D,Hu Y Y,Peng Y,Mao B G,Wang J J,Zhao B R.2013.Phenotypic difference and gene variation in agronomic traits between a rice variant with introduced exogenous genomic DNA and its receptor[J].Hybrid Rice,28(2):62-67.]
唐清杰,云勇,张吉贞,严小微,林尤珍,徐靖.2010.海南疣粒野生稻DNA导入栽培稻及其后代的SSR验证[J].亚热带植物科学,39(4):7-9.[Tang Q J,Yun Y,Zhang J J,Yan X W,Lin Y Z,Xu J.2010.Rice transformation by injecting Hainan Oryza meyeriana DNA and SSR detection[J].Subtropical Plant Science,39(4):7-9.]
王丰,柳武革,李金华,刘振荣,李曙光,廖亦龙,彭惠香,毛兴学.2004.水稻外源DNA导入系的创建及主要农艺性状分析[J].分子植物育种,2(4):489-494.[Wang F,Liu W G,Li J H,Liu Z R,Li S G,Liao Y L,Peng H X,Mao X X.2004.Construction of exogenous DNA introgression lines and analysis of its main traits in rice[J].Molecular Plant Breeding,2(4):489-494.]
王永斌,赵远玲,闫丽娟,猛庆用,王广金.2011.玉米DNA导入水稻变异后代的研究[J].中国农学通报,27(7):63-68.[Wang Y B,Zhao Y L,Yan L J,Meng Q Y,Wang GJ.2011.Study on variation offspring from different rice varieties introduced DNA of maize[J].Chinese Agricultural Science Bulletin,27(7):63-68.]
魏霞,李云峰,向佳,凌英华,杨正林,何光华,赵芳明.2015.干旱胁迫对导入高粱DNA的水稻变异系R21生理的影响[J].西南农业学报,28(3):930-934.[Wei X,Li Y F,Xiang J,Ling Y H,Yang Z L,He G H,Zhao F M.2015.Effect of drought stress on physiological characters of rice variant line R21 by introgression sorghum DNA[J].Southwest China Journal of Agricultural Sciences,28(3):930-934.]
赵鑫闻.2016.利用花粉管通道技术将外源银白杨DNA导入黑杨[J].植物学报,51(4):533-541.[Zhao X W.2016.Introduction of exogenous DNA of Populus alba into Sect.Aigeiros via pollen-tube pathway[J].Plant Biology,51(4):533-541.]
肇莹,肖军,杨镇,杨涛.2015.转导芦苇总DNA耐盐变异水稻SSH文库的构建及部分基因特异表达[J].江苏农业科学,43(3):13-16.[Zhao Y,Xiao J,Yang Z,Yang T.2015.Construction of SSH libraries and partial gene specific expression in variant rice with salt tolerance transduced from total DNA of reed[J].Jiangsu Agricultural Sciences,43(3):13-16.]
周光宇,翁坚,龚蓁蓁,曾以申,杨晚霞,沈慰芳,王自芬,陶全洲,黄骏麒,钱思颖,刘桂玲,应苗成,薛达元,洪爱华,徐英俊,陈善葆,段晓岚.1988.农业分子育种授粉后外源DNA导入植物的技术[J].中国农业科学,21(3):1-6.[Zhou G Y,Weng J,Gong Z Z,Zeng Y S,Yang W X,Shen W F,Wang Z F,Tao Q Z,Huang J Q,Qian S Y,Liu G L,Ying M C,Xue D Y,Hong A H,Xu Y J,Chen S B,Duan X L.1988.Molecular breeding of agriculture a technique for introducing exogenous DNA into plants after self pollination[J].Scientia Agricultura Sinica,21(3):1-6.]
周岩,游建,魏琦超,薛晓锋,何男男,马俊.2011.花粉管通道法转基因对小麦主要农艺性状的影响[J].河南科技学院学报,39(4):5-8.[Zhou Y,You J,Wei Q C,Xue X F,He N N,Ma J.2011.Study on the influence of agronomic traits of transformed wheat by pollen tube pathway[J].Journal of Henan Institute of Science and Technology,39(4):5-8.]
Luo Z X,Wu R.1989.A simple method for the transformation of rice via the pollen-tube pathway[J].Plant Moecular Bio1ogy Reporter,7(3):240.
Zhou G Y,Weng J,Zeng Y S,Huang J G,Qiao S Y,Liu GL.1983.Introduction of exogenous DNA into cotton embryos[J].Methods in Enzymology,101:433-481.