甘蓝型油菜的花期与生育期QTL定位
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摘要
【目的】对甘蓝型油菜花期和生育期QTL进行定位,为精细定位和克隆早花基因及开展分子标记辅助早熟油菜品种选育提供理论依据。【方法】以极早熟甘蓝型油菜G28、甘蓝型油菜H008及以二者为亲本构建的175个F1DH株系为材料,利用甘蓝型油菜60K SNP芯片分型技术绘制高密度遗传连锁图谱,并采用完备复合区间作图法对2016─2017年度丽江和临沧2个生长环境下甘蓝型油菜的花期(FT)和生育期(MT)田间调查数据进行QTL扫描分析。【结果】F1DH株系花期与生育期具有较明显的超亲现象,表明双亲材料控制花期和生育期的位点不同。F1DH株系在丽江生长环境下花期与生育期相关系数为0.63,在临沧生长环境下二者相关系数为0.79,即花期与生育期呈较高的正相关。利用SNP芯片构建的高密度遗传连锁图谱共包含19条连锁群,7601个SNPs位点,总长3838.2 c M。在丽江和临沧2个生长环境下共检测到6个花期QTL和5个生育期QTL,分布于A02、A07、C02、C03、C06、C07和C09连锁群上,可分别解释2.96%～17.40%和4.98%～11.82%的遗传变异。花期QTL qFTA02-1和qFTC03-2在2生长个环境下均可检测到,加性效应值相反,其中qFTA02-1具有最高的LOD值(20.43)、贡献率(17.40%)和加性效应值(3.27 d),且与生育期QTL qMTA02-1置信区间重叠,是最主要的花期主效QTL;qFTC03-2为次要的花期主效QTL。在qFTA02-1置信区间内发现2个拟南芥花期调控关键基因FLC和FY的油菜同源基因拷贝BnaA02g00370D和BnaA02g01670D。【结论】花期主效QTL qFTA02-1和qFTC03-2可用于分子标记辅助选育早熟油菜品种。BnaA02g00370D和BnaA02g01670D可能为qFTA02-1置信区间内控制甘蓝型油菜花期性状的目标基因。
【Objective】The study aimed to explore QTL mapping of flower time(FT)and maturity time(MT)of Brassica napus L. to obtain available information for finely mapping and cloning the gene related with early blooming trait and breeding of early-maturing rape variety assisted by molecular markers.【Method】G28,a super premature B. napus inbred line and H008,a B. napus inbred line,and 175 F1 DH lines with the above lines as parents were used as research materials.B. napus 60 K SNP array were used to construct a high density linkage map. The field survey data of FT and MT were collected from Lijiang and Lincang in 2016-2017. QTL identification of FT and MT was conducted using the model of complete composite interval mapping.【Result】The FT and MT of F1 DH line population presented an obvious transgressive segregation. It indicated that the loci that controlling FT and MT traits were different between their parents. Correlation coefficient of F1 DH line population between FT and MT was 0.63 in Lijiang and 0.79 in Lincang,indicating high positive correlation between FT and MT. High density linkage map established by SNP array contained 19 linkages,a total of7601 SNP sites covering 3838.2 cM. Six and five QTLs were detected related with FT and MT traits respectively in Lijiang and Lincang,distributing in A02,A07,C02,C03,C06,C07 and C09 linkages,which could explain 2.96%-17.40%and 4.98%-11.82% of the genetic effects respectively. The FT QTL,qFTA02-1 and q FTC03-2 were both detected in two environments,having opposite additive effects. The LOD value,contribution rate and additive effects of q FTA02-1 were as high as 20.43,17.40% and 3.27 d,respectively,which were all higher than those of qFTC03-2. In addition,the confidential interval of FT QTL qFTA02-1 overlapped with MT QTL qMTA02-1. q FTA02-1 was the major QTL at FT,and qFTC03-2 was the secondary major QTL at FT. Further analysis indicated that there were two candidate genes,BnaA02 g00370 D and BnaA02 g01670 D in confidential interval of qFTA02-1,which were homologous copies of FLC and FY genes controlling FT in Arabidopsis.【Conclusion】The major QTL at FT,qFTA02-1 and qFTC03-2,can be used in molecular markerassisted selection of early mature rape breeding,and the candidate genes BnaA02 g00370 D and BnaA02 g01670 D can be the target genes controlling the FT trait of B. napus in confidence interval of qFTA02-1.
【Objective】The study aimed to explore QTL mapping of flower time(FT)and maturity time(MT)of Brassica napus L. to obtain available information for finely mapping and cloning the gene related with early blooming trait and breeding of early-maturing rape variety assisted by molecular markers.【Method】G28,a super premature B. napus inbred line and H008,a B. napus inbred line,and 175 F1 DH lines with the above lines as parents were used as research materials.B. napus 60 K SNP array were used to construct a high density linkage map. The field survey data of FT and MT were collected from Lijiang and Lincang in 2016-2017. QTL identification of FT and MT was conducted using the model of complete composite interval mapping.【Result】The FT and MT of F1 DH line population presented an obvious transgressive segregation. It indicated that the loci that controlling FT and MT traits were different between their parents. Correlation coefficient of F1 DH line population between FT and MT was 0.63 in Lijiang and 0.79 in Lincang,indicating high positive correlation between FT and MT. High density linkage map established by SNP array contained 19 linkages,a total of7601 SNP sites covering 3838.2 cM. Six and five QTLs were detected related with FT and MT traits respectively in Lijiang and Lincang,distributing in A02,A07,C02,C03,C06,C07 and C09 linkages,which could explain 2.96%-17.40%and 4.98%-11.82% of the genetic effects respectively. The FT QTL,qFTA02-1 and q FTC03-2 were both detected in two environments,having opposite additive effects. The LOD value,contribution rate and additive effects of q FTA02-1 were as high as 20.43,17.40% and 3.27 d,respectively,which were all higher than those of qFTC03-2. In addition,the confidential interval of FT QTL qFTA02-1 overlapped with MT QTL qMTA02-1. q FTA02-1 was the major QTL at FT,and qFTC03-2 was the secondary major QTL at FT. Further analysis indicated that there were two candidate genes,BnaA02 g00370 D and BnaA02 g01670 D in confidential interval of qFTA02-1,which were homologous copies of FLC and FY genes controlling FT in Arabidopsis.【Conclusion】The major QTL at FT,qFTA02-1 and qFTC03-2,can be used in molecular markerassisted selection of early mature rape breeding,and the candidate genes BnaA02 g00370 D and BnaA02 g01670 D can be the target genes controlling the FT trait of B. napus in confidence interval of qFTA02-1.
引文
蔡东芳,张书芬,何俊平,朱家成,王建平,文雁成,曹金华,赵磊,王东国.2017.甘蓝型油菜抗旱机制及育种研究进展[J].河南农业科学,46(3):8-12.[Cai D F,Zhang S F,He J P,Zhu J C,Wang J P,Wen Y C,Cao J H,Zhao L,Wang D G.2017.Research progress on drought resistance mechanism and breeding in rapeseed(Brassica napus L.)[J].Journal of Henan Agricultural Sciences,46(3):8-12.]
陈盛,王宁宁,王玉康,陈松,曹维,李明,朱克明,彭琦,王政,谭小力.2017.一种快速高效筛选甘蓝型油菜转化植株的方法[J].江苏农业学报,33(5):982-985.[Chen S,Wang N N,Wang Y K,Chen S,Cao W,Li M,Zhu K M,Peng Q,Wang Z,Tan X L.2017.A rapid and efficient approach to screening transformed plants of Brassica napus[J].Jiangsu Journal of Agricultural Sciences,33(5):982-985.]
关周博,董育红,赵小光,韦世豪,郑磊,张耀文,田建华,赵卫国.2016.甘蓝型油菜生育期性状的遗传研究[J].上海农业学报,32(5):28-32.[Guan Z B,Dong Y H,Zhao XG,Wei S H,Zheng L,Zhang Y W,Tian J H,Zhao W G.2016.Genetic investigation on traits related to growth petiod in Brassica napus L[J].Acta Agriculturae Shanghai,32(5):28-32.]
黄吉祥,熊化鑫,潘兵,倪西源,张晓玉,赵坚义.2016.油菜开花期QTL定位及与粒重的遗传关联性[J].中国农业科学,49(16):3073-3083.[Huang J X,Xiong H X,Pan B,Ni X Y,Zhang X Y,Zhao J Y.2016.Mapping QTL of flowering time and their genetic relationships with seed weight in Brassica napus[J].Scientia Agricultura Sinica,49(16):3073-3083.]
姜成红,耿鑫鑫,魏文辉,姜慧芳.2017.甘蓝型油菜株高QTL定位及主效QTL区间候选基因预测[J].河南农业科学,46(8):27-31.[Jiang C H,Geng X X,Wei W H,Jiang HF.2017.QTL mapping of plant height and prediction of candidate genes in major QTL interval of Brassica napus[J].Journal of Henan Agricultural Sciences,46(8):27-31.]
李慧慧,张鲁燕,王建康.2010.数量性状基因定位研究中若干常见问题的分析与解答[J].作物学报,36(6):918-931.[Li H H,Zhang L Y,Wang J K.2010.Analysis and answers to frequently asked questions in quantitative trait locous mapping[J].Acta Agronomica Sinica,36(6):918-931.]
倪正斌,王陈燕,孙雪辉,吴昌庚,孙红芹,万林生,严国红.2018.甘蓝型油菜主要农艺性状相关性及主成分分析[J].江西农业学报,30(3):7-10.[Ni Z B,Wang C Y,Sun X H,Wu C G,Sun H Q,Wan L S,Yan G H.2018.Correlation and principal component analyses of main agronomic traits in Brassica napus[J].Acta Agriculturae Jiangxi,30(3):7-10.]
任照彬,王会涛,豆丹丹,任真真,柳华峰,库丽霞,陈彦惠.2018.玉米叶高点长等叶型性状QTL定位及上位性效应分析[J].河南农业大学学报,52(2):157-162.[Ren Z B,Wang H T,Dou D D,Ren Z Z,Liu H F,Ku L X,Chen Y H.2018.QTL mapping and epistatic effect analysis of high point long and other leaf shape related traits in maize[J].Journal of Henan Agricultural University,52(2):157-162.]
王钊辉,石超男,杨天啸,薛亚东,孙虎威,唐贵良,张战辉.2017.AtPHO2的CRISPR/Cas9靶向敲除及其在拟南芥中功能分析[J].河南农业大学学报,51(4):554-560.[Wang Z H,Shi C N,Yang T X,Xue Y D,Sun H W,Tang G L,Zhang Z H.2017.Gene knock out and functional analysis of AtPHO2 by CRISPR/Cas9-induced mutagenesis[J].Journal of Henan Agricultural University,51(4):554-560.]
徐文,安素妨,王艳,贾琳琳,鲁丹丹,张莹莹,刘建丰,李保全.2018.基于RNA-seq技术的甘蓝型油菜新转录本的鉴定[J].河南农业科学,47(4):38-42.[Xu W,An S F,Wang Y,Jia L L,Lu D D,Zhang Y Y,Liu J F,Li B Q.2018.Identification of novel transcripts of Brasscia napus by RNA-seq[J].Journal of Henan Agricultural Sciences,47(4):38-42.]
Feng W,Michaels S D.2011.Dual roles for FY in the regulation of FLC[J].Plant Signaling&Behavior,6(5):703-705.
Ferreira M E,Satagopan J,Yandell B S,Williams P H,Osborn T C.1995.Mapping loci controlling vernalization requirement and flowering time in Brassica napus[J].Theoretical Applied Genetics,90(5):727-732.
Helliwell C A,Wood C C,Robertson M,James Peacock W,Dennis E S.2006.The Arabidopsis FLC protein interacts directly in vivo with SOC1 and FT chromatin and is part of a high-molecular-weight protein complex[J].Plant Journal,46(2):183-192.
Hou J N,Long Y,Raman H,Zou X X,Wang J,Dai S T,Xiao Q Q,Li C,Fan L J,Liu B,Meng J L.2012.A touristlike MITE insertion in the upstream region of the BnFLC.A10 gene is associated with vernalization requirement in rapeseed(Brassica napus L.)[J].BMC Plant Biology,12:238.
Kim S Y,Park B S,Kwon S J,Kim J,Lim M H,Park Y D,Kim D Y,Suh S C,Jin Y M,Ahn J H,Lee Y H.2007.Delayed flowering time in Arabidopsis and Brassica rapa by the overexpression of FLOWERING LOCUS C(FLC)homologs isolated from Chinese cabbage(Brassica rapa L.ssp.pekinensis)[J].Plant Cell Reports,26(3):327-336.
Long Y,Shi J,Qiu D,Li R,Zhang C,Wang J,Hou J,Zhao J,Shi L,Park B S,Choi S R,Lim Y P,Meng J.2007.Flowering time quantitative trait Loci analysis of oilseed brassica in multiple environments and genomewide alignment with Arabidopsis[J].Genetics,177(4):2433-2444.
Meng L,Li H,Zhang L,Wang J.2015.QTL IciMapping.Integrated software for genetic linkage map construction and quantitative trait locus mapping in biparental populations[J].The Crop Journal,3(3):269-283.
Parkin I A,Gulden S M,Sharpe A G,Lukens L,Trick M,Osborn T C,Lydiate D J.2005.Segmental structure of the Brassica napus genome based on comparative analysis with Arabidopsis thaliana[J].Genetics,171(2):765-781.
Raman H,Dalton-Morgan J,Diffey S,Raman R,Alamery S,Edwards D,Batley J.2014.SNP markers-based map construction and genome-wide linkage analysis in Brassica napus[J].Plant Biotechnology Journal,12(7):851-860.
Sanda S L,Amasino R M.1996.Interaction of FLC and lateflowering mutations in Arabidopsis thaliana[J].Molecular and General Genetics,251(1):69-74.
Xu L P,Hu K N,Zhang Z Q,Guan C Y,Chen S,Hua W,Li J N,Wen J,Yi B,Shen J X,Ma C Z,Tu J X,Fu T D.2016.Genome-wide association study reveals the genetic architecture of flowering time in rapeseed(Brassica napus L.)[J].DNA Research,23(1):43-52.
Zhao J Y,Becker H C,Ding H D,Zhang Y F,Zhang D Q,Ecke W.2005.QTL of three agronomically important traits and their interactions with environment in a European×Chinese rapeseed population[J].Acta Genetica Sinica,32(9):969-978.
陈盛,王宁宁,王玉康,陈松,曹维,李明,朱克明,彭琦,王政,谭小力.2017.一种快速高效筛选甘蓝型油菜转化植株的方法[J].江苏农业学报,33(5):982-985.[Chen S,Wang N N,Wang Y K,Chen S,Cao W,Li M,Zhu K M,Peng Q,Wang Z,Tan X L.2017.A rapid and efficient approach to screening transformed plants of Brassica napus[J].Jiangsu Journal of Agricultural Sciences,33(5):982-985.]
关周博,董育红,赵小光,韦世豪,郑磊,张耀文,田建华,赵卫国.2016.甘蓝型油菜生育期性状的遗传研究[J].上海农业学报,32(5):28-32.[Guan Z B,Dong Y H,Zhao XG,Wei S H,Zheng L,Zhang Y W,Tian J H,Zhao W G.2016.Genetic investigation on traits related to growth petiod in Brassica napus L[J].Acta Agriculturae Shanghai,32(5):28-32.]
黄吉祥,熊化鑫,潘兵,倪西源,张晓玉,赵坚义.2016.油菜开花期QTL定位及与粒重的遗传关联性[J].中国农业科学,49(16):3073-3083.[Huang J X,Xiong H X,Pan B,Ni X Y,Zhang X Y,Zhao J Y.2016.Mapping QTL of flowering time and their genetic relationships with seed weight in Brassica napus[J].Scientia Agricultura Sinica,49(16):3073-3083.]
姜成红,耿鑫鑫,魏文辉,姜慧芳.2017.甘蓝型油菜株高QTL定位及主效QTL区间候选基因预测[J].河南农业科学,46(8):27-31.[Jiang C H,Geng X X,Wei W H,Jiang HF.2017.QTL mapping of plant height and prediction of candidate genes in major QTL interval of Brassica napus[J].Journal of Henan Agricultural Sciences,46(8):27-31.]
李慧慧,张鲁燕,王建康.2010.数量性状基因定位研究中若干常见问题的分析与解答[J].作物学报,36(6):918-931.[Li H H,Zhang L Y,Wang J K.2010.Analysis and answers to frequently asked questions in quantitative trait locous mapping[J].Acta Agronomica Sinica,36(6):918-931.]
倪正斌,王陈燕,孙雪辉,吴昌庚,孙红芹,万林生,严国红.2018.甘蓝型油菜主要农艺性状相关性及主成分分析[J].江西农业学报,30(3):7-10.[Ni Z B,Wang C Y,Sun X H,Wu C G,Sun H Q,Wan L S,Yan G H.2018.Correlation and principal component analyses of main agronomic traits in Brassica napus[J].Acta Agriculturae Jiangxi,30(3):7-10.]
任照彬,王会涛,豆丹丹,任真真,柳华峰,库丽霞,陈彦惠.2018.玉米叶高点长等叶型性状QTL定位及上位性效应分析[J].河南农业大学学报,52(2):157-162.[Ren Z B,Wang H T,Dou D D,Ren Z Z,Liu H F,Ku L X,Chen Y H.2018.QTL mapping and epistatic effect analysis of high point long and other leaf shape related traits in maize[J].Journal of Henan Agricultural University,52(2):157-162.]
王钊辉,石超男,杨天啸,薛亚东,孙虎威,唐贵良,张战辉.2017.AtPHO2的CRISPR/Cas9靶向敲除及其在拟南芥中功能分析[J].河南农业大学学报,51(4):554-560.[Wang Z H,Shi C N,Yang T X,Xue Y D,Sun H W,Tang G L,Zhang Z H.2017.Gene knock out and functional analysis of AtPHO2 by CRISPR/Cas9-induced mutagenesis[J].Journal of Henan Agricultural University,51(4):554-560.]
徐文,安素妨,王艳,贾琳琳,鲁丹丹,张莹莹,刘建丰,李保全.2018.基于RNA-seq技术的甘蓝型油菜新转录本的鉴定[J].河南农业科学,47(4):38-42.[Xu W,An S F,Wang Y,Jia L L,Lu D D,Zhang Y Y,Liu J F,Li B Q.2018.Identification of novel transcripts of Brasscia napus by RNA-seq[J].Journal of Henan Agricultural Sciences,47(4):38-42.]
Feng W,Michaels S D.2011.Dual roles for FY in the regulation of FLC[J].Plant Signaling&Behavior,6(5):703-705.
Ferreira M E,Satagopan J,Yandell B S,Williams P H,Osborn T C.1995.Mapping loci controlling vernalization requirement and flowering time in Brassica napus[J].Theoretical Applied Genetics,90(5):727-732.
Helliwell C A,Wood C C,Robertson M,James Peacock W,Dennis E S.2006.The Arabidopsis FLC protein interacts directly in vivo with SOC1 and FT chromatin and is part of a high-molecular-weight protein complex[J].Plant Journal,46(2):183-192.
Hou J N,Long Y,Raman H,Zou X X,Wang J,Dai S T,Xiao Q Q,Li C,Fan L J,Liu B,Meng J L.2012.A touristlike MITE insertion in the upstream region of the BnFLC.A10 gene is associated with vernalization requirement in rapeseed(Brassica napus L.)[J].BMC Plant Biology,12:238.
Kim S Y,Park B S,Kwon S J,Kim J,Lim M H,Park Y D,Kim D Y,Suh S C,Jin Y M,Ahn J H,Lee Y H.2007.Delayed flowering time in Arabidopsis and Brassica rapa by the overexpression of FLOWERING LOCUS C(FLC)homologs isolated from Chinese cabbage(Brassica rapa L.ssp.pekinensis)[J].Plant Cell Reports,26(3):327-336.
Long Y,Shi J,Qiu D,Li R,Zhang C,Wang J,Hou J,Zhao J,Shi L,Park B S,Choi S R,Lim Y P,Meng J.2007.Flowering time quantitative trait Loci analysis of oilseed brassica in multiple environments and genomewide alignment with Arabidopsis[J].Genetics,177(4):2433-2444.
Meng L,Li H,Zhang L,Wang J.2015.QTL IciMapping.Integrated software for genetic linkage map construction and quantitative trait locus mapping in biparental populations[J].The Crop Journal,3(3):269-283.
Parkin I A,Gulden S M,Sharpe A G,Lukens L,Trick M,Osborn T C,Lydiate D J.2005.Segmental structure of the Brassica napus genome based on comparative analysis with Arabidopsis thaliana[J].Genetics,171(2):765-781.
Raman H,Dalton-Morgan J,Diffey S,Raman R,Alamery S,Edwards D,Batley J.2014.SNP markers-based map construction and genome-wide linkage analysis in Brassica napus[J].Plant Biotechnology Journal,12(7):851-860.
Sanda S L,Amasino R M.1996.Interaction of FLC and lateflowering mutations in Arabidopsis thaliana[J].Molecular and General Genetics,251(1):69-74.
Xu L P,Hu K N,Zhang Z Q,Guan C Y,Chen S,Hua W,Li J N,Wen J,Yi B,Shen J X,Ma C Z,Tu J X,Fu T D.2016.Genome-wide association study reveals the genetic architecture of flowering time in rapeseed(Brassica napus L.)[J].DNA Research,23(1):43-52.
Zhao J Y,Becker H C,Ding H D,Zhang Y F,Zhang D Q,Ecke W.2005.QTL of three agronomically important traits and their interactions with environment in a European×Chinese rapeseed population[J].Acta Genetica Sinica,32(9):969-978.