紫斑百合远缘杂交亲和性分析及杂交障碍克服
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摘要
【目的】探究野生紫斑百合远缘杂交亲和性及克服杂交障碍的措施,为野生紫斑百合资源开发利用及具有绿色、清香、抗性等优良性状的百合新品种选育提供理论参考。【方法】以云南地区野生紫斑百合及栽培品系东方百合(7个品种)和OT百合(5个品种)为材料,采用切割花柱(CS)和常规授粉(NP)两种方法进行远缘杂交,分析紫斑百合与东方百合和OT百合的远缘杂交亲和性,通过荧光显微技术观察花粉萌发及花粉管生长动态,并通过染色体法鉴定F1代的倍性。【结果】常规授粉方式下,紫斑百合作母本的12个杂交组合中仅紫斑百合×索邦(NP)、紫斑百合×西诺红(NP)和紫斑百合×布卡迪(NP)3个杂交组合的亲和性相对较好,其他9个组合(NP)均杂交不亲和,存在受精前障碍,但切割花柱授粉方式下,紫斑百合作母本的12个杂交组合均具有不同程度亲和性,其中,紫斑百合×罗宾娜(CS)杂交亲和性最佳,其子房膨大率、有胚率、杂种胚萌发率和亲和指数分别为40.00%、15.12%、43.08%和6.05。以紫斑百合为父本的6个杂交组合在两种授粉方式下的杂交亲和性整体较差,仅切割花柱授粉方式下的罗宾娜×紫斑百合(CS)的子房膨大率和有胚率分别为20.00%和10.71%,其他5个杂交组合的子房假膨大或授粉后不久枯萎衰败。紫斑百合×索邦(CS)授粉后其花粉管到达子房胚珠的时间比紫斑百合×索邦(NP)提前4 d,有利于减少花粉管伸长过程中的营养消耗及杂交障碍的克服。西诺红×紫斑百合(NP)和黄天霸×紫斑百合(NP)2个杂交组合授粉8 h后花粉均在雌蕊柱头上萌发,花粉管沿着花柱向下延伸,黄天霸×紫斑百合(NP)的花粉萌发量较西诺红×紫斑百合(NP)少,但二者均在花柱和胚囊两端出现不同程度的胼胝反应,花粉管扭曲缠绕,阻碍花粉管伸长及伸向胚珠完成受精。紫斑百合×白色眼睛(CS)的杂交后代组培苗染色体数为24条,属二倍体;但紫斑百合×斯特腾(CS)和紫斑百合×维拉布兰斯(CS)的杂交后代组培苗染色体数分别为26和28条,为非整倍体植株。【结论】切割花柱授粉法可较好地克服紫斑百合与东方百合和OT百合远缘杂交受精前杂障碍,提高各杂交组合的子房膨大率、有胚率和杂种胚萌发率,且紫斑百合正反交组合的杂交亲和性差异明显,紫斑百合作母本的杂交组合较其作父本的杂交组合亲和性好,说明其杂交亲和性具有单向性。
【Objective】The study aimed to explore compatibility and the way to overcome the difficulties from distant hybridization of wild Lilium nepalense to provide a theoretical reference for the development of wild lily resources and the breeding of new lily varieties with good characteristics such as green,fragrance and resistance.【Method】Wild L. nepalense in Yunnan and cultivated lines Oriental hybrids(seven varieties),OT hybrids(five varieties)were used as experimental materials,pollination was carried out by cut-style method(CS)and normal pollination(NP)to analyze the distant hybridization compatibility between L. nepalense and Oriental hybrids,OT hybrids. Pollen germination and pollen tube growth were observed by fluorescence microscopy method,ploidy of F_1 hybrid progeny were identified by chromosome method.【Result】Under conventional pollination style,in the 12 combinations of interspecific hybridization which wild L.nepalense was as female parent,only three combinations of L. nepalense×Sorbonne(NP),L. nepalense×Viviana(NP)and L. nepalense×Bacardi(NP)had relatively good cross compatibility,but the other nine combinations(NP)with L. nepalense as female parent were all incompatible and had pre-fertilization obstacles. The 12 combinations by cutting style pollination all had compatibility with different levels. Among them,hybridization between L. nepalense and Robina was the best one,the ovary enlargement rate,embryo rate,germination rate and compatibility index of hybrid embryos were 40.00%,15.12%,43.08% and 6.05,respectively. The compatibility of six crossing combinations of L. nepalense as the male parent was poor under two pollination methods. Among the crossing combinations,only the ovary of crossing combination between Robina×L. nepalense(CS)by cutting style pollination developed,and the rate of ovary swelling and the rate of embryo developing were 20.00% and 10.71% respectively,but the ovaries of other five crossing combinations were pseudo-enlarged or withered shortly after pollination. When hybridization between L. nepalense as female parent and Sorbonne was used cutting style(CS)for pollination,the time of the pollen tube reached the ovule of the ovary which was 4 d earlier than normal pollination(NP). So cutting style method could reduce the nutrient consumption during the elongation of the pollen tube,which was beneficial to overcome the hybridization obstacle.Two combinations of distant hybridization Viviana×L. nepalense(NP)and Manissa×L. nepalense(NP)were carried out,the pollens of L. nepalense germinated on the stigma after 8 h of pollination,pollen germination of Manissa×L. nepalense(NP)was less than Viviana×L. nepalense(NP). But there were some abnormal phenomena such as winding and transverse growth during the pollen tube extending toward the style,and strong callus reaction happened in pollen tube and ovule bead hole,which hindered the growth of the pollen tube and reaching to the ovule for fertilization by fluorescence microscopic observation. The results from chromosome observation showed that hybrid between L. nepalense×White eyes(CS)was diploid with 24 chromosomes;while hybrids between L. nepalense×Stentor(CS),L. nepalense×Villa Balance(CS)were aneuploid with 26 and 28 chromosomes respectively.【Conclusion】Style-cutting pollination can overcome the pre-fertilization obstacles from distant hybridization between wild L. nepalense and Oriental hybrids,OT hybrids,and improve the ovary enlargement rate,embryonic rate and germination rate of each hybrid combination. There are great differences of compatibility in reciprocal crossing combinations when L. nepalense was used for parents. The combination with L. nepalense as female parent has better compatibility than that with L. nepalense as male parent. The results show that the hybridization compatibility is unidirectional.
【Objective】The study aimed to explore compatibility and the way to overcome the difficulties from distant hybridization of wild Lilium nepalense to provide a theoretical reference for the development of wild lily resources and the breeding of new lily varieties with good characteristics such as green,fragrance and resistance.【Method】Wild L. nepalense in Yunnan and cultivated lines Oriental hybrids(seven varieties),OT hybrids(five varieties)were used as experimental materials,pollination was carried out by cut-style method(CS)and normal pollination(NP)to analyze the distant hybridization compatibility between L. nepalense and Oriental hybrids,OT hybrids. Pollen germination and pollen tube growth were observed by fluorescence microscopy method,ploidy of F_1 hybrid progeny were identified by chromosome method.【Result】Under conventional pollination style,in the 12 combinations of interspecific hybridization which wild L.nepalense was as female parent,only three combinations of L. nepalense×Sorbonne(NP),L. nepalense×Viviana(NP)and L. nepalense×Bacardi(NP)had relatively good cross compatibility,but the other nine combinations(NP)with L. nepalense as female parent were all incompatible and had pre-fertilization obstacles. The 12 combinations by cutting style pollination all had compatibility with different levels. Among them,hybridization between L. nepalense and Robina was the best one,the ovary enlargement rate,embryo rate,germination rate and compatibility index of hybrid embryos were 40.00%,15.12%,43.08% and 6.05,respectively. The compatibility of six crossing combinations of L. nepalense as the male parent was poor under two pollination methods. Among the crossing combinations,only the ovary of crossing combination between Robina×L. nepalense(CS)by cutting style pollination developed,and the rate of ovary swelling and the rate of embryo developing were 20.00% and 10.71% respectively,but the ovaries of other five crossing combinations were pseudo-enlarged or withered shortly after pollination. When hybridization between L. nepalense as female parent and Sorbonne was used cutting style(CS)for pollination,the time of the pollen tube reached the ovule of the ovary which was 4 d earlier than normal pollination(NP). So cutting style method could reduce the nutrient consumption during the elongation of the pollen tube,which was beneficial to overcome the hybridization obstacle.Two combinations of distant hybridization Viviana×L. nepalense(NP)and Manissa×L. nepalense(NP)were carried out,the pollens of L. nepalense germinated on the stigma after 8 h of pollination,pollen germination of Manissa×L. nepalense(NP)was less than Viviana×L. nepalense(NP). But there were some abnormal phenomena such as winding and transverse growth during the pollen tube extending toward the style,and strong callus reaction happened in pollen tube and ovule bead hole,which hindered the growth of the pollen tube and reaching to the ovule for fertilization by fluorescence microscopic observation. The results from chromosome observation showed that hybrid between L. nepalense×White eyes(CS)was diploid with 24 chromosomes;while hybrids between L. nepalense×Stentor(CS),L. nepalense×Villa Balance(CS)were aneuploid with 26 and 28 chromosomes respectively.【Conclusion】Style-cutting pollination can overcome the pre-fertilization obstacles from distant hybridization between wild L. nepalense and Oriental hybrids,OT hybrids,and improve the ovary enlargement rate,embryonic rate and germination rate of each hybrid combination. There are great differences of compatibility in reciprocal crossing combinations when L. nepalense was used for parents. The combination with L. nepalense as female parent has better compatibility than that with L. nepalense as male parent. The results show that the hybridization compatibility is unidirectional.
引文
杜文文.2013.亚洲百合与铁炮百合杂交亲和性研究[D].武汉：华中农业大学.[Du W W.2013.Cross compatability between Asian Lily and Longiflorum Lily[D].Wuhan:Huazhong Agricultural University.]
傅立国，陈潭清，郎楷永，洪涛，林祁，李勇.2002.中国高等植物[M].青岛：青岛出版社.[Fu L G,Chen T Q,Lang KY,Hong T,Lin Q,Li Y.2002.Higher plants of China[M].Qingdao:Qingdao Publishing House.]
高雪，曹钦政，黄凤杰，贾桂霞.2015.亚洲百合与东方百合远缘杂交亲和性[J].东北林业大学学报，43(3):1-5.[Gao X,Cao Q Z,Huang F J,Jia G X.2015.Compatibility for lily distant hybridization between Asiatic hybrids and Oriental hybrids[J].Journal of Northeast Forestry University,43(3):1-5.]
何显静，李标，虞泓.2003.紫斑百合居群生物学研究[J].云南大学学报(自然科学版)，(S1):78-83.[He X J,Li B,Yu H.2003.The population study of Lilium nepalense[J].Journal of Agricultural University(Natural Science Edition),(S1):78-83.]
侯珺，罗建让，肖菲菲，张延龙，牛立新.2016.21种野生百合亲缘关系的ISSR分析[J].广西植物，36(9):1032-1038.[Hou J,Luo J R,Xiao F F,Zhang Y L,Niu L X.2016.Genetic relationship analysis of 21 wild species of Lilium by ISSR markers[J].Guihaia,36(9):1032-1038.]
雷家军，贺卫丽，赵艳.2014.不同授粉方式对亚洲百合与东方百合种间杂交结实的影响[J].东北农业大学学报，45(2):29-34.[Lei J J,He W L,Zhao Y.2014.Effect of different pollination methods on fruit setting of interspecific crosses between Asiatic and Oriental lily hybrids[J].Journal of Northeast Agricultural University,45(2):29-34.]
李标，虞泓，唐坤.2004.紫斑百合居群核型变异式样[J].重庆邮电学院学报(自然科学版)，16(1):98-102.[Li B,Yu H,Tang K.2004.Study on karyotypical variation in population of Lilium nepalense[J].Journal of Chongqing University(Natural Science Edition),16(1):98-102.]
李旦，罗一然，韩国伟，张雪，吴英英，何承忠.2017.野生紫斑百合多倍体诱导研究[J].云南农业大学学报(自然科学)，32(4):678-684.[Li D,Luo Y R,Han G W,Zhang X,Wu Y Y,He C Z.2017.Studies on polyploidy induction of wild Lilium nepalense D.Don[J].Journal of Yunnan Agricultural University(Natural Science),32(4):678-684.]
李婕，高亦珂，张启翔.2013.切割花柱法对百合杂交亲和性的影响[J].东北林业大学学报，41(5):98-101.[Li J,Gao Y K,Zhang Q X.2013.The effect of cut style methods on cross compatibility of lilies breeding[J].Journal of Northeast Forestry University,41(5):98-101.]
李守丽，石雷，张金政，龙雅宜.2006.百合育种研究进展[J].园艺学报，33(1):203-210.[Li S L,Shi L,Zhang J Z,Long Y Y.2006.Recent advances in lily breeding[J].Acta Horticulture Sinica,33(1):203-210.]
栗艳龄，梁建萍，罗凤霞.2012.切花百合杂种胚抢救方法[J].山西农业科学，40(6):566-570.[Li Y L,Liang J P,Luo F X.2012.Embryo rescue technology in the cut flower lily[J].Journal of Shanxi Agricultural Sciences,40(6):566-570.]
梁振旭.2014.川、陕及其毗邻地区野生百合种质资源调查与评价[D].杨凌：西北农林科技大学.[Liang Z X.2014.Investigation and evaluation on wild lilium native to Sichuan,Shanxi Province and its adjacent area[D].Yangling:Northwest A&F University.]
潘云兵，吴景芝，欧琳钧，车伟光，吴红芝.2018.野生百合与栽培百合远缘杂交亲和性的研究[J].云南农业大学学报(自然科学)，33(2):286-293.[Pan Y P,Wu J Z,Ou L J,Che W G,Wu H Z.2018.Studies on the compatibility of interspecific hybridization between wild lilies and cultivated lilies[J].Journal of Yunnan Agricultural University(Natural Science),33(2):286-293.]
王文和，王树栋，赵祥云，李小英，于建军，陈之欢.2007.百合远缘杂交花粉萌发及花粉管生长过程观察[J].西北植物学报，27(9):1790-1794.[Wang W H,Wang S D,Zhao X Y,Li X Y,Yu J J,Chen Z H.2007.Process of pollen sprouting and pollen tube growth on the lily distant hybridization[J].Acta Botanica Boreali-Occidentalia Sinica,27(9):1790-1794.]
王元军.2005.植物体内的胼胝质[J].生物学通报，40(1):18-19.[Wang Y J.2005.Callose in plants[J].Bulletin of Biology,40(1):18-19.]
吴英英，许昌慧，唐军荣，赵雁鸣，张雪娟，辛培尧，何承忠.2013.野生紫斑百合高效离体再生体系的建立[J].云南农业大学学报(自然科学)，28(2):242-246.[Wu Y Y,Xu C H,Tang J R,Zhao Y M,Zhang X J,Xin P Y,He CZ.2013.Establishment of an efficient micro-propagation system for wild Lilium nepalense D.Don[J].Journal of Yunnan Agricultural University(Natural Science),28(2):242-246.]
谢松林，王仙芝，牛立新，张延龙.2010.百合杂种系间杂交障碍的克服及3种幼胚离体培养方法的比较研究[J].西北植物学报，30(8):1573-1578.[Xie S L,Wang X Z,Niu L X,Zhang Y L.2010.Overcoming cross barriers and obtaining crossing hybrids between different hybrid groups of lily(Lilium)[J].Acta Botanica Boreali-Occidentalia Sinica,30(8):1573-1578.]
Barba-Gonzalez R,Lokker B H,Lim K B,Ramanna M S,van Tuyl J M.2004.Use of 2n gametes for the production of sexual polyploids from sterile Oriental×Asiatic hybrids of lilies(Lilium)[J].Theoretical and Applied Gene-tics,109(6):1125-1132.
Lim K B,van Tuyl J M.2006.Lily[M].Dordrecht:Springer Netherlands.
Okazaki K,Asano Y,Oosawa K.1994.Interspecifc hybrids between Lilium-Oriental hybrid and L-Asiatic hybrid produced by embryo culture with revised media[J].Breed Science 44:59-64.
Rhee H K,Kim K S.2008.Interspecific hybridization and polyploidization in lily breeding[J].Acta Horticulturae,766:441-446.
van Tuyl J M,Arens P,Ramanna M S,Shahin A,Khan N,Xie S,Marasek-Ciolakowska A,Lim K B,Barba-Gonzalez R.2011.Lilium[M].Berlin,Heidelberg:Springer:161-183.
van Tuyl J M,Arens P,Shahin A,Marasek-Cio?akowska A,Barba-Gonzalez R,Kim H T,Lim K B.2018.Lilium[M].Cham:Springer International Publishing:481-512.
van Tuyl J M,van Di?n M P,Van Creij M G M,van Kleinwee T C M,Franken J,Bino R J.1991.Application of in vitro pollination,ovary culture,ovule culture and embryo rescue for overcoming incongruity barriers in interspecific Lilium crosses[J].Plant Science,74(1):115-126.
van Tuyl J M,van Dijken H S,Chi H S,Lim K B.2000.Breakthroughs in interspecific hybridization of lily[J].Acta Horticulturae,508:83-88.
傅立国，陈潭清，郎楷永，洪涛，林祁，李勇.2002.中国高等植物[M].青岛：青岛出版社.[Fu L G,Chen T Q,Lang KY,Hong T,Lin Q,Li Y.2002.Higher plants of China[M].Qingdao:Qingdao Publishing House.]
高雪，曹钦政，黄凤杰，贾桂霞.2015.亚洲百合与东方百合远缘杂交亲和性[J].东北林业大学学报，43(3):1-5.[Gao X,Cao Q Z,Huang F J,Jia G X.2015.Compatibility for lily distant hybridization between Asiatic hybrids and Oriental hybrids[J].Journal of Northeast Forestry University,43(3):1-5.]
何显静，李标，虞泓.2003.紫斑百合居群生物学研究[J].云南大学学报(自然科学版)，(S1):78-83.[He X J,Li B,Yu H.2003.The population study of Lilium nepalense[J].Journal of Agricultural University(Natural Science Edition),(S1):78-83.]
侯珺，罗建让，肖菲菲，张延龙，牛立新.2016.21种野生百合亲缘关系的ISSR分析[J].广西植物，36(9):1032-1038.[Hou J,Luo J R,Xiao F F,Zhang Y L,Niu L X.2016.Genetic relationship analysis of 21 wild species of Lilium by ISSR markers[J].Guihaia,36(9):1032-1038.]
雷家军，贺卫丽，赵艳.2014.不同授粉方式对亚洲百合与东方百合种间杂交结实的影响[J].东北农业大学学报，45(2):29-34.[Lei J J,He W L,Zhao Y.2014.Effect of different pollination methods on fruit setting of interspecific crosses between Asiatic and Oriental lily hybrids[J].Journal of Northeast Agricultural University,45(2):29-34.]
李标，虞泓，唐坤.2004.紫斑百合居群核型变异式样[J].重庆邮电学院学报(自然科学版)，16(1):98-102.[Li B,Yu H,Tang K.2004.Study on karyotypical variation in population of Lilium nepalense[J].Journal of Chongqing University(Natural Science Edition),16(1):98-102.]
李旦，罗一然，韩国伟，张雪，吴英英，何承忠.2017.野生紫斑百合多倍体诱导研究[J].云南农业大学学报(自然科学)，32(4):678-684.[Li D,Luo Y R,Han G W,Zhang X,Wu Y Y,He C Z.2017.Studies on polyploidy induction of wild Lilium nepalense D.Don[J].Journal of Yunnan Agricultural University(Natural Science),32(4):678-684.]
李婕，高亦珂，张启翔.2013.切割花柱法对百合杂交亲和性的影响[J].东北林业大学学报，41(5):98-101.[Li J,Gao Y K,Zhang Q X.2013.The effect of cut style methods on cross compatibility of lilies breeding[J].Journal of Northeast Forestry University,41(5):98-101.]
李守丽，石雷，张金政，龙雅宜.2006.百合育种研究进展[J].园艺学报，33(1):203-210.[Li S L,Shi L,Zhang J Z,Long Y Y.2006.Recent advances in lily breeding[J].Acta Horticulture Sinica,33(1):203-210.]
栗艳龄，梁建萍，罗凤霞.2012.切花百合杂种胚抢救方法[J].山西农业科学，40(6):566-570.[Li Y L,Liang J P,Luo F X.2012.Embryo rescue technology in the cut flower lily[J].Journal of Shanxi Agricultural Sciences,40(6):566-570.]
梁振旭.2014.川、陕及其毗邻地区野生百合种质资源调查与评价[D].杨凌：西北农林科技大学.[Liang Z X.2014.Investigation and evaluation on wild lilium native to Sichuan,Shanxi Province and its adjacent area[D].Yangling:Northwest A&F University.]
潘云兵，吴景芝，欧琳钧，车伟光，吴红芝.2018.野生百合与栽培百合远缘杂交亲和性的研究[J].云南农业大学学报(自然科学)，33(2):286-293.[Pan Y P,Wu J Z,Ou L J,Che W G,Wu H Z.2018.Studies on the compatibility of interspecific hybridization between wild lilies and cultivated lilies[J].Journal of Yunnan Agricultural University(Natural Science),33(2):286-293.]
王文和，王树栋，赵祥云，李小英，于建军，陈之欢.2007.百合远缘杂交花粉萌发及花粉管生长过程观察[J].西北植物学报，27(9):1790-1794.[Wang W H,Wang S D,Zhao X Y,Li X Y,Yu J J,Chen Z H.2007.Process of pollen sprouting and pollen tube growth on the lily distant hybridization[J].Acta Botanica Boreali-Occidentalia Sinica,27(9):1790-1794.]
王元军.2005.植物体内的胼胝质[J].生物学通报，40(1):18-19.[Wang Y J.2005.Callose in plants[J].Bulletin of Biology,40(1):18-19.]
吴英英，许昌慧，唐军荣，赵雁鸣，张雪娟，辛培尧，何承忠.2013.野生紫斑百合高效离体再生体系的建立[J].云南农业大学学报(自然科学)，28(2):242-246.[Wu Y Y,Xu C H,Tang J R,Zhao Y M,Zhang X J,Xin P Y,He CZ.2013.Establishment of an efficient micro-propagation system for wild Lilium nepalense D.Don[J].Journal of Yunnan Agricultural University(Natural Science),28(2):242-246.]
谢松林，王仙芝，牛立新，张延龙.2010.百合杂种系间杂交障碍的克服及3种幼胚离体培养方法的比较研究[J].西北植物学报，30(8):1573-1578.[Xie S L,Wang X Z,Niu L X,Zhang Y L.2010.Overcoming cross barriers and obtaining crossing hybrids between different hybrid groups of lily(Lilium)[J].Acta Botanica Boreali-Occidentalia Sinica,30(8):1573-1578.]
Barba-Gonzalez R,Lokker B H,Lim K B,Ramanna M S,van Tuyl J M.2004.Use of 2n gametes for the production of sexual polyploids from sterile Oriental×Asiatic hybrids of lilies(Lilium)[J].Theoretical and Applied Gene-tics,109(6):1125-1132.
Lim K B,van Tuyl J M.2006.Lily[M].Dordrecht:Springer Netherlands.
Okazaki K,Asano Y,Oosawa K.1994.Interspecifc hybrids between Lilium-Oriental hybrid and L-Asiatic hybrid produced by embryo culture with revised media[J].Breed Science 44:59-64.
Rhee H K,Kim K S.2008.Interspecific hybridization and polyploidization in lily breeding[J].Acta Horticulturae,766:441-446.
van Tuyl J M,Arens P,Ramanna M S,Shahin A,Khan N,Xie S,Marasek-Ciolakowska A,Lim K B,Barba-Gonzalez R.2011.Lilium[M].Berlin,Heidelberg:Springer:161-183.
van Tuyl J M,Arens P,Shahin A,Marasek-Cio?akowska A,Barba-Gonzalez R,Kim H T,Lim K B.2018.Lilium[M].Cham:Springer International Publishing:481-512.
van Tuyl J M,van Di?n M P,Van Creij M G M,van Kleinwee T C M,Franken J,Bino R J.1991.Application of in vitro pollination,ovary culture,ovule culture and embryo rescue for overcoming incongruity barriers in interspecific Lilium crosses[J].Plant Science,74(1):115-126.
van Tuyl J M,van Dijken H S,Chi H S,Lim K B.2000.Breakthroughs in interspecific hybridization of lily[J].Acta Horticulturae,508:83-88.
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