甘蔗与河八王杂交后代染色体传递及河八王血缘在后代中的遗传
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摘要
为探明甘蔗野生基因在杂交过程中的遗传情况,提高甘蔗野生种质资源的利用效率,本研究利用染色体计数法及AFLP结合荧光毛细管电泳技术,开展了甘蔗与河八王杂交的染色体传递和亲本特异位点在杂交种中的遗传分析。结果表明,甘蔗×河八王组合的F1染色体传递行为有n+n和n+2n方式,BC1是n+n方式;利用AFLP标记结合毛细管电泳技术开展了河八王野生基因在F1、BC1后代中的遗传分析,并利用24对AFLP引物组合对4个河八王F1后代、9个BC1后代及其双亲进行了扩增,共检测到总条带数1 774条,其中多态性条带1 690条,多态性检出率为95.26%,各引物组合多态性变幅为87.04%～100%。进一步分析表明,母本桂糖05-3256的特异位点传递给F1后代的比例是63.92%～71.65%,父本广西河八王1特异位点传递给F1后代的比例是8.66%～12.85%。母本崖城94-46特异位点传递给BC1后代的比例是35.19%～48.50%,父本T6-3特异位点传递给BC1后代的比例是33.97%～59.33%;广西河八王1特异位点传递到BC1后代时,比例仅为1.12%～3.35%。另外,F1、BC1的特异位点保持了较高水平。本研究说明了河八王作为父本进行杂交利用过程中,其野生基因在后代中丢失严重,在BC1已处于很低水平,建议前期将河八王作为母本杂交利用,可能会聚合更多有益的河八王野生血缘,从而改良甘蔗的抗逆性。
In order to find out the inheritance of wild genes in sugarcane during hybridization and improve the utilization efficiency of wild sugarcane germplasm resources, genetic analysis of chromosome transmission and parent specific loci in hybrid between sugarcane and Narenga porphyrocoma were carried out by using chromosome counting and AFLP combined with fluorescence capillary electrophoresis technology. The results showed that thechromosomal transfer models were "n +n" and "n +2n" in F1 generation of Sugarcane ×Narenga porphyrocoma combination, and "n +n" mode in BC1population; 24 pairs of AFLP primer combinations were used for amplification for 4 individuals of F1 generation, 9 individuals of BC1 generation and their parents. A total of 1 774 bands were detected, of which 1 690 were polymorphic and the polymorphic rate was 95.26%. The polymorphism of each primer combination ranged from 87.04% to 100%. Further analysis showed that the proportion of the specific loci transferred from the maternal parent GT05-3256 to individuals in F1 generation ranged from 63.92%to 71.65%, and the ratio of the specific loci transferred from the paternal parent GXN1(Narenga porphyrocoma) to individuals in F1 generation ranged from 8.66% to 12.85%. The proportion of specific loci transferred from the maternal parental YC94-46 to individuals in BC1 generation ranged from 35.19% to 48.50%, and the proportion of specific loci transferred from the paternal parent T6-3 to individuals in BC1 generation ranged from 33.97% to59.33%. The ratio of specific loci in GXN1 inherited to individuals in BC1 population, the ratio ranged from 1.12%to 3.35% only. In addition, the specific loci of F1 and BC1remained relatively high. This study showed that the wild genes of Narenga porphyrocoma were seriously lost in the offspring and were at a very low level in BC1 generation during the process of crossbreeding which used Narenga porphyrocoma as the male parent. It is suggested that the crossbreeding of Narenga porphyrocoma as female parent in the early stage might gather more beneficial wild blood relationship, and might improve the stress resistance of sugarcane.
In order to find out the inheritance of wild genes in sugarcane during hybridization and improve the utilization efficiency of wild sugarcane germplasm resources, genetic analysis of chromosome transmission and parent specific loci in hybrid between sugarcane and Narenga porphyrocoma were carried out by using chromosome counting and AFLP combined with fluorescence capillary electrophoresis technology. The results showed that thechromosomal transfer models were "n +n" and "n +2n" in F1 generation of Sugarcane ×Narenga porphyrocoma combination, and "n +n" mode in BC1population; 24 pairs of AFLP primer combinations were used for amplification for 4 individuals of F1 generation, 9 individuals of BC1 generation and their parents. A total of 1 774 bands were detected, of which 1 690 were polymorphic and the polymorphic rate was 95.26%. The polymorphism of each primer combination ranged from 87.04% to 100%. Further analysis showed that the proportion of the specific loci transferred from the maternal parent GT05-3256 to individuals in F1 generation ranged from 63.92%to 71.65%, and the ratio of the specific loci transferred from the paternal parent GXN1(Narenga porphyrocoma) to individuals in F1 generation ranged from 8.66% to 12.85%. The proportion of specific loci transferred from the maternal parental YC94-46 to individuals in BC1 generation ranged from 35.19% to 48.50%, and the proportion of specific loci transferred from the paternal parent T6-3 to individuals in BC1 generation ranged from 33.97% to59.33%. The ratio of specific loci in GXN1 inherited to individuals in BC1 population, the ratio ranged from 1.12%to 3.35% only. In addition, the specific loci of F1 and BC1remained relatively high. This study showed that the wild genes of Narenga porphyrocoma were seriously lost in the offspring and were at a very low level in BC1 generation during the process of crossbreeding which used Narenga porphyrocoma as the male parent. It is suggested that the crossbreeding of Narenga porphyrocoma as female parent in the early stage might gather more beneficial wild blood relationship, and might improve the stress resistance of sugarcane.
引文
Bian X.,Dong L.H.,Sun Y.F.,Tao L.A.,Zhu J.R.,Zhou Q.M.,Yang L.H.,An R.D.,Lang R.B.,Yu H.X.,Feng W.,and Jing Y.F.,2014,Principal component analysis of drought resistance related traits of Saccharum spontaneum L.and its F1hybrids,Ganhan Diqu Nongye Yanjiu(Agricultural Research in the Acid Areas),32(3):56-61(边芯,董立华,孙有芳,桃联安,朱建荣,周清明,杨李和,安汝东,郎荣斌,俞华先,冯蔚,经艳芬,2014,云南割手密及其血缘F1代材料抗旱相关性状的主成分分析,干旱地区农业研究,32(3):56-61)
Cao Z.Q.,Xiao F.R.,Chen S.Y.,He L.L.,and Li F.S.,2017,Identification of cold tolerance in seven wil d sugarcane and three offsprings at seedling stage,Zuowu Zazhi(Crops),(5):43-48(曹哲群,肖芙荣,陈疏影,何丽莲,李富生,2017,7个蔗茅野生种及其后代材料苗期耐寒性鉴定,作物杂志,(5):43-48)
Chen Y.,Wang Z.Q.,Fu Z.W.,Yang Y.T.,Guo J.L.,Su Y.C.,and Xu L.P.,2015,Cloning and expression analysis of metallothionein gene ScMT2-1-4 in sugarcane,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(2):357-364(陈云,王竹青,傅志伟,杨玉婷,郭晋隆,苏亚春,许莉萍,2015,甘蔗金属硫蛋白基因(ScMT2-1-4)的克隆及表达分析,基因组学与应用生物学,34(2):357-364)
Hang Y.J.,Fu C.,Lin W.L.,Liu S.M.,Gao J.H.,Deng Z.H.,Huang Z.X.,Lin Y.Q.,and Chen R.K.,2015,Genetic analysis of chromosome in 3 BC1clones from the distant crossing between Saccharum spp.and Erianthus arundinaceus,Redai Zuowu Xuebao(Chinese Journal of Tropical Crops),36(1):53-58(黄永吉,符成,林炜乐,刘少谋,高嘉慧,邓祖湖,黄忠兴,林彦铨,陈如凯,2015,3个甘蔗与斑茅远缘杂交后代BC1的染色体遗传分析,热带作物学报,36(1):53-58)
Huang Y.X.,Luo T.,Lin X.Q.,Zhang B.Q.,Zhou S.,Yang C.F.,Lei J.C.,Gao Y.J.,Duan W.X.,Zhang G.M.,and Li Y.R.,2017,GISH analysis of intergeneric complex between Erianthus arundinaceus and Saccharum spontaneum(GXAS07-6-1)and its F1hybrids from crosses with sugarcane(Saccharum spp.),Zhiwu Yichuan Ziyuan Xuebao(Journal of Plant Genetic Resources),18(3):461-466(黄玉新,罗霆,林秀琴,张保青,周珊,杨翠芳,雷敬超,高轶静,段维兴,张革民,李杨瑞,2017,斑茅割手密复合体(GXAS07-6-1)及其与甘蔗F1的GISH分析,植物遗传资源学报,18(3):461-466)
Hunag Y.X.,Luo T.,Gao Y.J.,Zhang B.Q.,Zhou S.,Yang C.F.,Duan W.X.,Lei J.C.,Wei J.J.,Zhang G.M.,and Li Y.R.,2016,Authenticity identification and chromosomes genetic analysis of backcross progeny between sugarcane(Saccharum officinarum)and intergeneric hybrid complex(Erianthus arundinaceus×Saccharum spontaneum),Nanfang Nongye Xuebao(Journal of Southern Agriculture),47(10):1642-1647(黄玉新,罗霆,高轶静,张保青,周珊,杨翠芳,段维兴,雷敬超,韦金菊,张革民,李杨瑞,2016,甘蔗与斑茅割手密复合体的回交后代真实性鉴定及染色体遗传分析,南方农业学报,47(10):1642-1647)
Li X.J.,Liu H.B.,Lin X.Q.,Wu Z.D.,Xu C.H.,and Liu X.L.,2015,In silico cloning and bioinformatics analysis of KNOXgene in sugarcane(Sckn1),Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(1):136-142(李旭娟,刘洪博,林秀琴,吴转娣,徐超华,刘新龙,2015,甘蔗KNOX基因(Sckn1)的电子克隆及生物信息学分析,基因组学与应用生物学,34(1):136-142)
Lin X.Q.,Lu X.,Liu X.L.,Mao J.,Zi Q.Y.,Liu H.B.,Li Z.L.,Li X.J.,Xu C.H.,Li C.J.,and Cai Q.,2017,Analysis of intergeneric hybrids(F1,BC1)of Saccharum×Erianthusrockii using morphology and GISH technique,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),15(6):2307-2316(林秀琴,陆鑫,刘新龙,毛钧,字秋艳,刘洪博,李忠磊,李旭娟,徐超华,李纯佳,蔡青,2017,甘蔗与滇蔗茅杂交后代F1,BC1表型和GISH分析,分子植物育种,15(6):2307-2316)
Liu S.M.,Fu C.,and Chen Y.S.,2007,Utilization of Erianthus arundinaceus progeny through backcross over the last decade at Hainan sugarcane breeding station,Ganzhe Tangye(Sugarcane and Canesugar),(2):1-6(刘少谋,符成,陈勇生,2007,近十年海南甘蔗育种场斑茅后代回交利用研究,甘蔗糖业,(2):1-6)
Liu X.H.,Fang F.X.,Gao Y.J.,Zhang R.H.,Song H.Z.,Yang R.Z.,Fang W.K.,Duan W.X.,Luo T.,Zhang G.M.,and Li Y.R.,2012a,Identification and genetic analysis of hybrid from cross between Erianthus arundinacius(Retz.)Jesws.and Saccharum spontaneum L.,Zuowu Xuebao(Acta Agronomica Sinica),38(5):914-920(刘昔辉,方锋学,高轶静,张荣华,宋焕忠,杨荣仲,方位宽,段维兴,罗霆,张革民,李杨瑞,2012a,斑茅割手密杂种后代真实性鉴定及遗传分析,作物学报,38(5):914-920)
Liu X.H.,Fang F.X.,Zhang R.H.,Song H.Z.,Yang R.Z.,Gao Y.J.,Lei J.C.,Luo T.,Duan W.X.,Zhang G.M.,and Li Y.R.,2012b,Identification of progenies from sugarcane×Narenga porphyrocoma(Hance)Bor by SSR marker,Xinan Nongye Xuebao(Southwest China Journal of Agricural Sciences),25(1):38-42(刘昔辉,方锋学,张荣华,宋焕忠,杨荣仲,高轶静,雷敬超,罗霆,段维兴,张革民,李杨瑞,2012b,甘蔗与河八王属间杂种的SSR标记鉴定,西南农业学报,25(1):38-42)
Liu X.H.,Song H.Z.,Zhang G.M.,Duan W.X.,Zhang R.H.,and Li Y.R.,2017,Phenotypic variation and genetic diversity in the collections of Erianthus arundinaceus(Retz.)Jesw.,Sugar Tech.,19(4):359-367
Lu X.,Mao J.,Liu H.B.,Liu X.L.,Ma L.,Su H.S.,and Cai Q.,2012,Innovation germplasm of Erianthus rockiiⅠ.Establishment of F1intergeneric distant hybridization of Saccharum officinarum L.and Erianthus rockii and the identification of F1by SSR molecular markers,Zhiwu Yichuan Ziyuan X-uebao(Journal of Plant Genetic Resources),13(2):321-324(陆鑫,毛钧,刘洪博,刘新龙,马丽,苏火生,蔡青,2012,甘蔗野生种滇蔗茅种质创新利用研究Ⅰ.甘蔗与滇蔗茅远缘杂交F1群体构建与SSR分子标记鉴定,植物遗传资源学报,13(2):321-324)
Wang X.H.,Li F.S.,He L.L.,Lou H.B.,Yang Q.H.,and He S.C.,2014,Chromosomal transmission of F2,BC1progenies between sugarcane and Erianthus fulvus,Zhongguo Nongxue Tongbao(Chinese Agricultural Science Bulletin),30(15):81-85(王先宏,李富生,何丽莲,娄红波,杨清辉,何顺长,2014,甘蔗与蔗茅杂交F2,BC1代的染色体遗传分析,中国农学通报,30(15):81-85)
Zheng X.F.,Zhang M.Q.,Li Q.W.,Lao F.Y.,Deng Z.H.,Chen R.K.,and Yang Y.H.,2004,Utilization and characterisation of the genuine intergeneric hybrids from the cross of Saccharum and E.arundinaceum,(2):molecular identification of genuine hybrids from the cross of Saccharum and E.arundinaceum,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),2(1):35-42(郑雪芳,张木清,李奇伟,劳方业,邓祖湖,陈如凯,杨业后,2004,甘蔗斑茅的杂交利用及其杂种后代鉴定系列研究(二):甘蔗斑茅远缘真实杂种的分子鉴定,分子植物育种,2(1):35-42)
Cao Z.Q.,Xiao F.R.,Chen S.Y.,He L.L.,and Li F.S.,2017,Identification of cold tolerance in seven wil d sugarcane and three offsprings at seedling stage,Zuowu Zazhi(Crops),(5):43-48(曹哲群,肖芙荣,陈疏影,何丽莲,李富生,2017,7个蔗茅野生种及其后代材料苗期耐寒性鉴定,作物杂志,(5):43-48)
Chen Y.,Wang Z.Q.,Fu Z.W.,Yang Y.T.,Guo J.L.,Su Y.C.,and Xu L.P.,2015,Cloning and expression analysis of metallothionein gene ScMT2-1-4 in sugarcane,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(2):357-364(陈云,王竹青,傅志伟,杨玉婷,郭晋隆,苏亚春,许莉萍,2015,甘蔗金属硫蛋白基因(ScMT2-1-4)的克隆及表达分析,基因组学与应用生物学,34(2):357-364)
Hang Y.J.,Fu C.,Lin W.L.,Liu S.M.,Gao J.H.,Deng Z.H.,Huang Z.X.,Lin Y.Q.,and Chen R.K.,2015,Genetic analysis of chromosome in 3 BC1clones from the distant crossing between Saccharum spp.and Erianthus arundinaceus,Redai Zuowu Xuebao(Chinese Journal of Tropical Crops),36(1):53-58(黄永吉,符成,林炜乐,刘少谋,高嘉慧,邓祖湖,黄忠兴,林彦铨,陈如凯,2015,3个甘蔗与斑茅远缘杂交后代BC1的染色体遗传分析,热带作物学报,36(1):53-58)
Huang Y.X.,Luo T.,Lin X.Q.,Zhang B.Q.,Zhou S.,Yang C.F.,Lei J.C.,Gao Y.J.,Duan W.X.,Zhang G.M.,and Li Y.R.,2017,GISH analysis of intergeneric complex between Erianthus arundinaceus and Saccharum spontaneum(GXAS07-6-1)and its F1hybrids from crosses with sugarcane(Saccharum spp.),Zhiwu Yichuan Ziyuan Xuebao(Journal of Plant Genetic Resources),18(3):461-466(黄玉新,罗霆,林秀琴,张保青,周珊,杨翠芳,雷敬超,高轶静,段维兴,张革民,李杨瑞,2017,斑茅割手密复合体(GXAS07-6-1)及其与甘蔗F1的GISH分析,植物遗传资源学报,18(3):461-466)
Hunag Y.X.,Luo T.,Gao Y.J.,Zhang B.Q.,Zhou S.,Yang C.F.,Duan W.X.,Lei J.C.,Wei J.J.,Zhang G.M.,and Li Y.R.,2016,Authenticity identification and chromosomes genetic analysis of backcross progeny between sugarcane(Saccharum officinarum)and intergeneric hybrid complex(Erianthus arundinaceus×Saccharum spontaneum),Nanfang Nongye Xuebao(Journal of Southern Agriculture),47(10):1642-1647(黄玉新,罗霆,高轶静,张保青,周珊,杨翠芳,段维兴,雷敬超,韦金菊,张革民,李杨瑞,2016,甘蔗与斑茅割手密复合体的回交后代真实性鉴定及染色体遗传分析,南方农业学报,47(10):1642-1647)
Li X.J.,Liu H.B.,Lin X.Q.,Wu Z.D.,Xu C.H.,and Liu X.L.,2015,In silico cloning and bioinformatics analysis of KNOXgene in sugarcane(Sckn1),Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(1):136-142(李旭娟,刘洪博,林秀琴,吴转娣,徐超华,刘新龙,2015,甘蔗KNOX基因(Sckn1)的电子克隆及生物信息学分析,基因组学与应用生物学,34(1):136-142)
Lin X.Q.,Lu X.,Liu X.L.,Mao J.,Zi Q.Y.,Liu H.B.,Li Z.L.,Li X.J.,Xu C.H.,Li C.J.,and Cai Q.,2017,Analysis of intergeneric hybrids(F1,BC1)of Saccharum×Erianthusrockii using morphology and GISH technique,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),15(6):2307-2316(林秀琴,陆鑫,刘新龙,毛钧,字秋艳,刘洪博,李忠磊,李旭娟,徐超华,李纯佳,蔡青,2017,甘蔗与滇蔗茅杂交后代F1,BC1表型和GISH分析,分子植物育种,15(6):2307-2316)
Liu S.M.,Fu C.,and Chen Y.S.,2007,Utilization of Erianthus arundinaceus progeny through backcross over the last decade at Hainan sugarcane breeding station,Ganzhe Tangye(Sugarcane and Canesugar),(2):1-6(刘少谋,符成,陈勇生,2007,近十年海南甘蔗育种场斑茅后代回交利用研究,甘蔗糖业,(2):1-6)
Liu X.H.,Fang F.X.,Gao Y.J.,Zhang R.H.,Song H.Z.,Yang R.Z.,Fang W.K.,Duan W.X.,Luo T.,Zhang G.M.,and Li Y.R.,2012a,Identification and genetic analysis of hybrid from cross between Erianthus arundinacius(Retz.)Jesws.and Saccharum spontaneum L.,Zuowu Xuebao(Acta Agronomica Sinica),38(5):914-920(刘昔辉,方锋学,高轶静,张荣华,宋焕忠,杨荣仲,方位宽,段维兴,罗霆,张革民,李杨瑞,2012a,斑茅割手密杂种后代真实性鉴定及遗传分析,作物学报,38(5):914-920)
Liu X.H.,Fang F.X.,Zhang R.H.,Song H.Z.,Yang R.Z.,Gao Y.J.,Lei J.C.,Luo T.,Duan W.X.,Zhang G.M.,and Li Y.R.,2012b,Identification of progenies from sugarcane×Narenga porphyrocoma(Hance)Bor by SSR marker,Xinan Nongye Xuebao(Southwest China Journal of Agricural Sciences),25(1):38-42(刘昔辉,方锋学,张荣华,宋焕忠,杨荣仲,高轶静,雷敬超,罗霆,段维兴,张革民,李杨瑞,2012b,甘蔗与河八王属间杂种的SSR标记鉴定,西南农业学报,25(1):38-42)
Liu X.H.,Song H.Z.,Zhang G.M.,Duan W.X.,Zhang R.H.,and Li Y.R.,2017,Phenotypic variation and genetic diversity in the collections of Erianthus arundinaceus(Retz.)Jesw.,Sugar Tech.,19(4):359-367
Lu X.,Mao J.,Liu H.B.,Liu X.L.,Ma L.,Su H.S.,and Cai Q.,2012,Innovation germplasm of Erianthus rockiiⅠ.Establishment of F1intergeneric distant hybridization of Saccharum officinarum L.and Erianthus rockii and the identification of F1by SSR molecular markers,Zhiwu Yichuan Ziyuan X-uebao(Journal of Plant Genetic Resources),13(2):321-324(陆鑫,毛钧,刘洪博,刘新龙,马丽,苏火生,蔡青,2012,甘蔗野生种滇蔗茅种质创新利用研究Ⅰ.甘蔗与滇蔗茅远缘杂交F1群体构建与SSR分子标记鉴定,植物遗传资源学报,13(2):321-324)
Wang X.H.,Li F.S.,He L.L.,Lou H.B.,Yang Q.H.,and He S.C.,2014,Chromosomal transmission of F2,BC1progenies between sugarcane and Erianthus fulvus,Zhongguo Nongxue Tongbao(Chinese Agricultural Science Bulletin),30(15):81-85(王先宏,李富生,何丽莲,娄红波,杨清辉,何顺长,2014,甘蔗与蔗茅杂交F2,BC1代的染色体遗传分析,中国农学通报,30(15):81-85)
Zheng X.F.,Zhang M.Q.,Li Q.W.,Lao F.Y.,Deng Z.H.,Chen R.K.,and Yang Y.H.,2004,Utilization and characterisation of the genuine intergeneric hybrids from the cross of Saccharum and E.arundinaceum,(2):molecular identification of genuine hybrids from the cross of Saccharum and E.arundinaceum,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),2(1):35-42(郑雪芳,张木清,李奇伟,劳方业,邓祖湖,陈如凯,杨业后,2004,甘蔗斑茅的杂交利用及其杂种后代鉴定系列研究(二):甘蔗斑茅远缘真实杂种的分子鉴定,分子植物育种,2(1):35-42)