萝卜地方优良品种高制种潜力雄性不育系转育方法研究
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摘要
不育系的制种潜力是决定制种产量的重要因素。在前期选育过程中对不育系制种潜力关注不够,往往导致所选育的品种制种产量低甚至无法制种,造成品种无法推广。我国拥有丰富的优良地方品种萝卜资源,但是存在着种质退化、栽培面积缩减、栽培区域小而分散的问题。培育杂交种是提升优良地方品种品质、维持栽培面积和扩大栽培地域的有效途径。本研究将Ogura胞质转育到26份优良地方品种萝卜资源,发现其中16份种质资源在Ogura胞质中的不育率达到或接近100%;建立了一种基于网纱隔离和壁蜂授粉的鉴定方法,用于分析这16份优良地方品种萝卜资源的保持系及其Ogura不育系的制种产量与潜力,发现练丝萝卜和五月红两份资源的不育系和保持系都具有较高的角果产量,有望转育成高制种潜力的不育系亲本,红灯笼、大青皮和心里美3份资源中都分离出具有较高种子产量的胞质雄性不育单株,但是其保持系种子产量很低,需要提高保持系的种子产量;小红袍水萝卜和扶沟萝卜的不育系和保持系的种子产量都处于中等水平,具有优化和应用的潜力;红水萝卜、青皮脆等5份资源不适合Ogura胞质不育系制种;狗头罐、绛县白、象牙白3份资源是自交不亲和材料。本研究发现,Ogura胞质的不同品种以及同一品种不同单株间制种产量差异巨大,说明从转育初代开始逐代监测和筛选制种潜力是非常必要的。本研究介绍一种简易高效鉴定及筛选萝卜胞质不育系制种潜力的方法,并用此方法初步筛选出一批高制种潜力的萝卜地方品种Ogura胞质雄性不育材料。
The seed production potential of male sterile lines is the main factor affecting hybrid seed productivity. The lack of early selection method always leads to low hybrid seed yield and unsuitable for application. A collection of radish landraces with high quality have been developed in China over decades,but nowadays these germplasms showed characterization declining,cultivation area reducing and scattering.Hybrid breeding is an effective way to improve the agronomic quality,maintain the cultivation area and expand the planting region of the landraces. By hybridization using the Ogura cytoplasm with 26 radish landraces,16 introgression lines were absolute or near absolute male sterile. A osmia pollination and nylon mesh isolation method were used to test the seed production potential of the male sterile and maintainer lines of these 16 landraces. The two accessions,Lian-si and May-Red,showed high silique yield in both male sterile and maintainer plants,with the potential of developing high-seed-yielding male sterile lines. The Red-lanterns,Biggreen-skin and Beautiful-in-heart contain high silique yield plants in male sterile lines while all maintainer plants showed low yield. Improving the seed productivity of maintainer lines are needed for these three landraces. The Little-red-robe summer radish and Fu-gou radish produced medium quantity of siliques from both male sterile and maintainer plants,have application potential if substantial selection can improve the seed yield. The Red summer radish,Green crisp and three other landraces were proved not suitable to produce hybrid seeds use Ogura-CMS due to low yield. The Dog-Canister,Jiang-xian-white and Ivory were identi?ed as self incompatibility materials.This study revealed that the seed yield are differing signi?cantly among varieties and single plants,indicating a necessary of selecting the high-yielding plant descendants with generations. Thus,this worked reported a simple and efficient method for screening the seed production potential,which finally resulted in generation of seven Ogura-CMS radishes with high seed yielding potential.
The seed production potential of male sterile lines is the main factor affecting hybrid seed productivity. The lack of early selection method always leads to low hybrid seed yield and unsuitable for application. A collection of radish landraces with high quality have been developed in China over decades,but nowadays these germplasms showed characterization declining,cultivation area reducing and scattering.Hybrid breeding is an effective way to improve the agronomic quality,maintain the cultivation area and expand the planting region of the landraces. By hybridization using the Ogura cytoplasm with 26 radish landraces,16 introgression lines were absolute or near absolute male sterile. A osmia pollination and nylon mesh isolation method were used to test the seed production potential of the male sterile and maintainer lines of these 16 landraces. The two accessions,Lian-si and May-Red,showed high silique yield in both male sterile and maintainer plants,with the potential of developing high-seed-yielding male sterile lines. The Red-lanterns,Biggreen-skin and Beautiful-in-heart contain high silique yield plants in male sterile lines while all maintainer plants showed low yield. Improving the seed productivity of maintainer lines are needed for these three landraces. The Little-red-robe summer radish and Fu-gou radish produced medium quantity of siliques from both male sterile and maintainer plants,have application potential if substantial selection can improve the seed yield. The Red summer radish,Green crisp and three other landraces were proved not suitable to produce hybrid seeds use Ogura-CMS due to low yield. The Dog-Canister,Jiang-xian-white and Ivory were identi?ed as self incompatibility materials.This study revealed that the seed yield are differing signi?cantly among varieties and single plants,indicating a necessary of selecting the high-yielding plant descendants with generations. Thus,this worked reported a simple and efficient method for screening the seed production potential,which finally resulted in generation of seven Ogura-CMS radishes with high seed yielding potential.
引文
[1]王方艳·我国萝卜产业发展问题及对策·中国农机化学报,2016, 37(11):188-191Wang F Y. Problems and countermeasures of radish industry development in China-Journal of Chinese Agricultural Mechanization, 2016,37(11):188-191
[2]孔秋生,李锡香,向长萍,王海平,宋江萍·萝卜种质资源亲缘关系的RAPD分析.植物遗传资源学报,2004, 5(2):156-160,169Kong Q S, Li X X, Xiang C P, Wang H P, Song J P.Analysis on the phylogenetic relationship of radish(Raphanus sativus L-)germplasm resources with RAPD-Journal of Plant Genetic Resources, 2004, 5(2):156-160, 169
[3]孔秋生,李锡香,向长萍,邱杨,沈镝·栽培萝卜种质亲缘关系的AFLP分析·中国农业科学,2005,38(5):1017-1023Kong Q S, Li X X, Xiang C P, Qiu Y, Shen D-Phylogenetic analysis of cultivated radish(Raphanus sativus L-)germplasm with AFLP. Scientia Agricultura Sinica, 2005, 38(5):1017-1023
[4]邱杨,李锡香,李清霞,陈亦辰,沈镝,王海平,宋江萍·利用SSR标记构建萝卜种质资源分子身份证·植物遗传资源学报,2014, 15(3):648-654Qiu Y, Li X X, Li Q X, Chen Y C, Shen D, Wang H P, Song J P-Establishment of the molecular identification for radish germplasm using SSR markers. Journal of Plant Genetic Resources, 2014, 15(3):648-654
[5]李红双,李锡香,沈镝,杨宇红,邱杨,王海平,龚慧芝·萝卜优异种质对芜菁花叶病毒抗性的遗传分析·植物遗传资源学报,2010, 11(2):152-156LiHS,Li X X,Shen D,Yang Y H,Qiu Y,Wang H P,Gong H Z.Genetic analysis of the resistance to TuMV in elite radish germplasm.Journal of Plant Genetic Resources, 2010, 11(2):152-156
[6]杨好慧,邱杨,宋江萍,张晓辉,王鸿娜,王超楠,王海平,李锡香·萝卜种质资源对芜菁花叶病毒的抗性鉴定与评价·植物遗传资源学报,2017, 18(5):810-818Yang H H, Qiu Y, Song J P, Zhang X H, Wang H N, Wang C N, Wang H P, Li X X-Identification and evaluation for the resistance of radish(Raphanus sativus L.)germplasm resources to turnip mosaic virus. Journal of Plant Genetic Resources,2017, 18(5):810-818
[7]李红双,李锡香·萝卜抗源抗黑腐病性状的遗传分析·北京:中国园艺学会十字花科分会第七届学术研讨会,2009Li H S, Li X X.Genetic analysisi of black rot resistance in radish resistant resource-Beijing:The 7th Symposium of the Chinese Horticultural Society Crucifer Branch, 2009
[8]段韫丹,邱杨,汪精磊,王海平,张晓辉,沈镝,宋江萍,李锡香·萝卜不同抗源对黑腐病抗性的遗传分析·植物遗传资源学报,2015, 16(1):1-6Duan Y D, Qiu Y, Wang J L, Wang H P, Zhang X H, Shen D, Song J P, Li X X. Genetic analysis on the resistance of different radish germplasm to black rot. Journal of Plant Genetic Resources,2015, 16(1):1-6
[9]杨好慧,原玉香,张晓辉,魏小春,邱杨,宋江萍,吴春晖,段蒙蒙,李锡香·萝卜种质资源对根肿病抗性的鉴定·园艺学报,2017,44(S1):2534Yang H H, Yuan Y X, Zhang X H, Wei X C, Qiu Y, Song J P,Wu C H, Duan M M, Li X X. Identification of radish germplasm resources resistance to clubroot disease-Acta Horticulturae Sinica,2017, 44(S1):2534
[10]张素君,邱杨,宋江萍,王海平,张晓辉,沈镝,李锡香·萝卜种质资源耐抽薹性鉴定评价.植物遗传资源学报,2014, 15(2):262-269Zhang S J, Qiu Y, Song J P, Wang H P, Zhang X H, Shen D, Li X X. Identification for the tolerance to bolting of radish germplasm resources. Journal of Plant Genetic Resources,2014, 15(2):262-269
[11]华贝贝,邱杨,段韫丹,崔娜,张晓辉,沈镝,宋江萍,李锡香.萝卜(Raphanus sativus L.)种质莱菔子素含量分析与评价·植物遗传资源学报,2013, 14(6):1038-1044Hua B B, Qiu Y, Duan Y D, Cui N, Zhang X H, Shen D, Song J P, Li X X. Analysis and evaluation sulforaphene content in radish(Raphanus sativus L.)germplasm resources. Journal of Plant Genetic Resources, 2013, 14(6):1038-1044
[12]李晓琪,李锡香,罗越华,邱杨,沈镝,王海平·萝卜自交不亲和基因SLG6的CAPS标记的开发·植物遗传资源学报,2009, 10(1):118-125Li X Q, Li X X, Luo Y H, Qiu Y, Shen D, Wang H P. CAPS marker for self-incompatible gene SLG6 in Raphanus sativusJournal of Plant Genetic Resources, 2009, 10(1):118-125
[13] Bohra A,Jha U C,Adhimoolam P·Cytoplasmic male sterility(CMS)in hybrid breeding in field crops. Plant Cell Reports,2016, 35(5):967-993
[14] Ogura H-Studies on the new male-sterility in Japanese radish with special references to the utilization of this sterility towards the special raising of hybrid seeds·Mern Fac Agricultural Kagoshima Universily, 1968,6(2):39-78
[15] Iwabuchi M, Koizuka N, Fujimoto H, Sakai T, Imamura J.Identification and expression of the kosena radish(Raphanus sativus cv. Kosena)homologue of the ogura radish CMS-associated gene, orf138. Plant Molecular Biology, 1999,39(1):183-188
[16] Park J Y,Lee Y P,Lee J,Choi B S,Kim S,Yang T J. Complete mitochondrial genome sequence and identification of a candidate gene responsible for cytoplasmic male sterility in radish(Raphanus sativus L.)containing DCGMS cytoplasm. Theoretical and Applied Genetics, 2013, 126(7):1763-1774
[17] Duroc Y, Hiard S, Vrielynck N, Ragu S, Budar F. The Ogura sterility-inducing protein forms a large complex without interfering with the oxidative phosphorylation components in rapeseed mitochondria. Plant Molecular Biology, 2009, 70:123-137
[18]Desloire S,Gherbi H,Laloui W,Marhadour S,Clouet V,Cattolico L,Falentin C,Giancola S,Renard M,Budar F,Small,Caboche M,Delourme R, Bendahmane A. Identification of the fertility restoration locus,Rfo, in radish, as a member of the pentatricopeptide-repeat protein family. EMBO Reports, 2003,4(6):588-594
[19] Brown G G, Formanova N, Jin H, Wargachuk R, Dendy C,Patil P, Laforest M, Zhang J, Cheung W Y, Landry B S. The radish Rfo restorer gene of Ogura cytoplasmic male sterility encodes a protein with multiple pentatricopeptide repeats. Plant Journal for Cell and Molecular Biology, 2003,35(2):262-272
[20]Koizuka N, Imai R, Fujimoto H, Hayakawa T, Kimura Y,Kohno-Murase J, Sakai T, Kawasaki S, Imamura J. Genetic characterization of a pentatricopeptide repeat protein gene,orf687, that restores fertility in the cytoplasmic male-sterile Kosena radish. Plant Journal, 2003,34(4):407-415
[21]Wang Z W,Wang C,Gao L,Lei G,Shi Y,Yuan Z,Chang P X,Ting W. Heterozygous alleles restore male fertility to cytoplasmic male-sterile radish(Raphanus sativus L.):a case of overdominance.Journal of Experimental Botany, 2013,64(7):2041
[22] Menczel L, Morgan A, Brown S, Maliga P. Fusion-mediated combination of Ogura-type cytoplasmic male sterility with Brassica napus plastids using X-irradiated CMS protoplasts.Plant Cell Reports, 1987,6(2):98-101
[23]Jarl C I,van Grinsven M Q,van den Mark F. Correction of chlorophyll-defective male-sterile winter oilseed rape(Brassica napus)through organelle exchange:molecular analysis of the cytoplasm of parental lines and corrected progeny. Theoretical and Applied Genetics, 1989, 77(1):135-141
[24] Kirti P B, Banga S S, Prakash S, Chopra V L. Transfer of Ogu cytoplasmic male sterility to Brassica juncea and improvement of the male sterile line through somatic cell fusion. Theoretical Applied Genetic, 1995,91(3):517-521
[25]徐立功,韩太利.十字花科蔬菜亲本制种采用壁蜂授粉的实用技术.中国蔬菜,2014(9):74-76Xu L G, Han T L. Practical techniques for pollination of cruciferous vegetable parents using wall bee pollination. China Vegetables, 2014(9):74-76
[26]张丽·王庆彪·萝卜新品种京红5号的选育.中国蔬菜,2017(7):73-75Zhang L, Wang Q B-A New rasish F1 hybrid-'Jinghong No-5'·China Vegetables, 2017(7):73-75
[27]张丽,王庆彪.水果萝卜新品种“京脆2号”的选育.北方园艺,2017(10):156-158Zhang L, Wang Q B. Breeding of a new fruit-type radish hybrid'Jingcui No.2'Northern Horticulture,2017(10):156-158
[28]郎丰庆,王施慧,刘淑梅,侯丽霞·秋萝卜天正秋红1号及其栽培技术要点·中国蔬菜,2017(4):102-103Lang F Q, Wang S H, Li S M, Hou L X, Autumn radish Tianzheng qiuhong No. 1 and its cultivation techniques. China Vegetables,2017(4):102-103
[2]孔秋生,李锡香,向长萍,王海平,宋江萍·萝卜种质资源亲缘关系的RAPD分析.植物遗传资源学报,2004, 5(2):156-160,169Kong Q S, Li X X, Xiang C P, Wang H P, Song J P.Analysis on the phylogenetic relationship of radish(Raphanus sativus L-)germplasm resources with RAPD-Journal of Plant Genetic Resources, 2004, 5(2):156-160, 169
[3]孔秋生,李锡香,向长萍,邱杨,沈镝·栽培萝卜种质亲缘关系的AFLP分析·中国农业科学,2005,38(5):1017-1023Kong Q S, Li X X, Xiang C P, Qiu Y, Shen D-Phylogenetic analysis of cultivated radish(Raphanus sativus L-)germplasm with AFLP. Scientia Agricultura Sinica, 2005, 38(5):1017-1023
[4]邱杨,李锡香,李清霞,陈亦辰,沈镝,王海平,宋江萍·利用SSR标记构建萝卜种质资源分子身份证·植物遗传资源学报,2014, 15(3):648-654Qiu Y, Li X X, Li Q X, Chen Y C, Shen D, Wang H P, Song J P-Establishment of the molecular identification for radish germplasm using SSR markers. Journal of Plant Genetic Resources, 2014, 15(3):648-654
[5]李红双,李锡香,沈镝,杨宇红,邱杨,王海平,龚慧芝·萝卜优异种质对芜菁花叶病毒抗性的遗传分析·植物遗传资源学报,2010, 11(2):152-156LiHS,Li X X,Shen D,Yang Y H,Qiu Y,Wang H P,Gong H Z.Genetic analysis of the resistance to TuMV in elite radish germplasm.Journal of Plant Genetic Resources, 2010, 11(2):152-156
[6]杨好慧,邱杨,宋江萍,张晓辉,王鸿娜,王超楠,王海平,李锡香·萝卜种质资源对芜菁花叶病毒的抗性鉴定与评价·植物遗传资源学报,2017, 18(5):810-818Yang H H, Qiu Y, Song J P, Zhang X H, Wang H N, Wang C N, Wang H P, Li X X-Identification and evaluation for the resistance of radish(Raphanus sativus L.)germplasm resources to turnip mosaic virus. Journal of Plant Genetic Resources,2017, 18(5):810-818
[7]李红双,李锡香·萝卜抗源抗黑腐病性状的遗传分析·北京:中国园艺学会十字花科分会第七届学术研讨会,2009Li H S, Li X X.Genetic analysisi of black rot resistance in radish resistant resource-Beijing:The 7th Symposium of the Chinese Horticultural Society Crucifer Branch, 2009
[8]段韫丹,邱杨,汪精磊,王海平,张晓辉,沈镝,宋江萍,李锡香·萝卜不同抗源对黑腐病抗性的遗传分析·植物遗传资源学报,2015, 16(1):1-6Duan Y D, Qiu Y, Wang J L, Wang H P, Zhang X H, Shen D, Song J P, Li X X. Genetic analysis on the resistance of different radish germplasm to black rot. Journal of Plant Genetic Resources,2015, 16(1):1-6
[9]杨好慧,原玉香,张晓辉,魏小春,邱杨,宋江萍,吴春晖,段蒙蒙,李锡香·萝卜种质资源对根肿病抗性的鉴定·园艺学报,2017,44(S1):2534Yang H H, Yuan Y X, Zhang X H, Wei X C, Qiu Y, Song J P,Wu C H, Duan M M, Li X X. Identification of radish germplasm resources resistance to clubroot disease-Acta Horticulturae Sinica,2017, 44(S1):2534
[10]张素君,邱杨,宋江萍,王海平,张晓辉,沈镝,李锡香·萝卜种质资源耐抽薹性鉴定评价.植物遗传资源学报,2014, 15(2):262-269Zhang S J, Qiu Y, Song J P, Wang H P, Zhang X H, Shen D, Li X X. Identification for the tolerance to bolting of radish germplasm resources. Journal of Plant Genetic Resources,2014, 15(2):262-269
[11]华贝贝,邱杨,段韫丹,崔娜,张晓辉,沈镝,宋江萍,李锡香.萝卜(Raphanus sativus L.)种质莱菔子素含量分析与评价·植物遗传资源学报,2013, 14(6):1038-1044Hua B B, Qiu Y, Duan Y D, Cui N, Zhang X H, Shen D, Song J P, Li X X. Analysis and evaluation sulforaphene content in radish(Raphanus sativus L.)germplasm resources. Journal of Plant Genetic Resources, 2013, 14(6):1038-1044
[12]李晓琪,李锡香,罗越华,邱杨,沈镝,王海平·萝卜自交不亲和基因SLG6的CAPS标记的开发·植物遗传资源学报,2009, 10(1):118-125Li X Q, Li X X, Luo Y H, Qiu Y, Shen D, Wang H P. CAPS marker for self-incompatible gene SLG6 in Raphanus sativusJournal of Plant Genetic Resources, 2009, 10(1):118-125
[13] Bohra A,Jha U C,Adhimoolam P·Cytoplasmic male sterility(CMS)in hybrid breeding in field crops. Plant Cell Reports,2016, 35(5):967-993
[14] Ogura H-Studies on the new male-sterility in Japanese radish with special references to the utilization of this sterility towards the special raising of hybrid seeds·Mern Fac Agricultural Kagoshima Universily, 1968,6(2):39-78
[15] Iwabuchi M, Koizuka N, Fujimoto H, Sakai T, Imamura J.Identification and expression of the kosena radish(Raphanus sativus cv. Kosena)homologue of the ogura radish CMS-associated gene, orf138. Plant Molecular Biology, 1999,39(1):183-188
[16] Park J Y,Lee Y P,Lee J,Choi B S,Kim S,Yang T J. Complete mitochondrial genome sequence and identification of a candidate gene responsible for cytoplasmic male sterility in radish(Raphanus sativus L.)containing DCGMS cytoplasm. Theoretical and Applied Genetics, 2013, 126(7):1763-1774
[17] Duroc Y, Hiard S, Vrielynck N, Ragu S, Budar F. The Ogura sterility-inducing protein forms a large complex without interfering with the oxidative phosphorylation components in rapeseed mitochondria. Plant Molecular Biology, 2009, 70:123-137
[18]Desloire S,Gherbi H,Laloui W,Marhadour S,Clouet V,Cattolico L,Falentin C,Giancola S,Renard M,Budar F,Small,Caboche M,Delourme R, Bendahmane A. Identification of the fertility restoration locus,Rfo, in radish, as a member of the pentatricopeptide-repeat protein family. EMBO Reports, 2003,4(6):588-594
[19] Brown G G, Formanova N, Jin H, Wargachuk R, Dendy C,Patil P, Laforest M, Zhang J, Cheung W Y, Landry B S. The radish Rfo restorer gene of Ogura cytoplasmic male sterility encodes a protein with multiple pentatricopeptide repeats. Plant Journal for Cell and Molecular Biology, 2003,35(2):262-272
[20]Koizuka N, Imai R, Fujimoto H, Hayakawa T, Kimura Y,Kohno-Murase J, Sakai T, Kawasaki S, Imamura J. Genetic characterization of a pentatricopeptide repeat protein gene,orf687, that restores fertility in the cytoplasmic male-sterile Kosena radish. Plant Journal, 2003,34(4):407-415
[21]Wang Z W,Wang C,Gao L,Lei G,Shi Y,Yuan Z,Chang P X,Ting W. Heterozygous alleles restore male fertility to cytoplasmic male-sterile radish(Raphanus sativus L.):a case of overdominance.Journal of Experimental Botany, 2013,64(7):2041
[22] Menczel L, Morgan A, Brown S, Maliga P. Fusion-mediated combination of Ogura-type cytoplasmic male sterility with Brassica napus plastids using X-irradiated CMS protoplasts.Plant Cell Reports, 1987,6(2):98-101
[23]Jarl C I,van Grinsven M Q,van den Mark F. Correction of chlorophyll-defective male-sterile winter oilseed rape(Brassica napus)through organelle exchange:molecular analysis of the cytoplasm of parental lines and corrected progeny. Theoretical and Applied Genetics, 1989, 77(1):135-141
[24] Kirti P B, Banga S S, Prakash S, Chopra V L. Transfer of Ogu cytoplasmic male sterility to Brassica juncea and improvement of the male sterile line through somatic cell fusion. Theoretical Applied Genetic, 1995,91(3):517-521
[25]徐立功,韩太利.十字花科蔬菜亲本制种采用壁蜂授粉的实用技术.中国蔬菜,2014(9):74-76Xu L G, Han T L. Practical techniques for pollination of cruciferous vegetable parents using wall bee pollination. China Vegetables, 2014(9):74-76
[26]张丽·王庆彪·萝卜新品种京红5号的选育.中国蔬菜,2017(7):73-75Zhang L, Wang Q B-A New rasish F1 hybrid-'Jinghong No-5'·China Vegetables, 2017(7):73-75
[27]张丽,王庆彪.水果萝卜新品种“京脆2号”的选育.北方园艺,2017(10):156-158Zhang L, Wang Q B. Breeding of a new fruit-type radish hybrid'Jingcui No.2'Northern Horticulture,2017(10):156-158
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