亚洲棉与拟似棉远缘杂种的合成与鉴定

doi:10.13560/j.cnki.biotech.bull.1985.2019-1249

摘要/Abstract

摘要： 以亚洲棉为母本,野生种拟似棉为父本进行远缘杂交,人工辅助授粉合成杂种F₁,验证F₁杂种的真实性,以期进一步加倍成异源四倍体新种质。采用重复授粉和赤霉素保铃等措施提高杂交结铃率,对F₁进行形态学及SSR分子标记鉴定。结果表明,F₁杂种有典型的合子后生殖隔离现象,植株培养过程中伴随有杂种致死,F₁杂种幼苗叶型大部分介于双亲之间且整体偏向于父本拟似棉;SSR分子标记结果显示杂种F₁不仅扩增出双亲的互补带,还出现了双亲没有的新带;经过统计分析发现杂种F₁中总的遗传成分比例母本占45.91%,父本占40.98%,新出现的组合带占13.11%。从分子水平证明了杂种的真实性,同时伴随的杂交过程也发生了AD基因组互作及遗传重组。

关键词: 亚洲棉, 拟似棉, 远缘杂交, 生殖隔离, SSR

Abstract: Gossypium arboreum as a female parent had distant hybrid with Gossypium gossypioides as male parent. Artificial pollination was used to synthesize hybrid F₁,and the authenticity of the F₁ hybrids was verified,aiming to further doubling it into a new allotetraploid germplasm. Repeated pollination and gibberellin boll keeping measures were used to improve the boll setting rate of hybrid,and the F₁ was identified by morphology and SSR molecular markers. The results showed that the F₁ hybrids had typical post-zygotic reproductive isolation,and the hybrids died in the process of plant culture. Most leaf types of the F₁ hybrid seedlings were amid parents and tended to resemble male parents as a whole. The results of SSR molecular markers demonstrated that the hybrid F₁ not only amplified complementary bands of parents,but also new bands that parents did not have appeared. Statistical analysis revealed that the total genetic components of the hybrid F₁ accounted for 45.91% of the female parents,40.98% of the male parents and 13.11% of new combination bands. The study not only proves the authenticity of hybrids at the molecular level,but also shows that AD genome interaction and genetic recombination occur along with the hybridization process.

Key words: Gossypium arboreum, Gossypium gossypioides, distant hybridization, reproductive isolation, SSR

王一帆, 郑赟, 荣二花, 吴玉香. 亚洲棉与拟似棉远缘杂种的合成与鉴定[J]. 生物技术通报, 2020, 36(8): 1-7.

WANG Yi-fan, ZHENG Yun, RONG Er-hua, WU Yu-xiang. Synthesis and Identification of Distant Hybrids Between Gossypium arboreum and Gossypium gossypioides[J]. Biotechnology Bulletin, 2020, 36(8): 1-7.

参考文献

[1] Fryxell PA.A revised taxonomic interpretation of Gosspium L. (Malvaceae)[J]. Pheedea, 1992, 2(2):108-165.
[2] Wendel JF, Grover CE.Taxonomy and evolution of the cotton genus, Gossypium[M]. Fang DD, Percy RG. Cotton American Society of Agronomy Inc., Madison:2015.
[3] 冀丽霞, 郭宝德, 雷伟, 等. 棉花特异种质材料的选育研究[J]. 种子科技, 2017, 35(8):126-128.
Ji LX,Guo BD, Lei W, et al.Study on breeding of cotton specific cermplasm materials[J]. Seed Scicence, 2017, 35(8):126-128.
[4] 郭旺珍, 王凯, 张天真. 利用SSR标记技术研究棉属A、D染色体组的进化[J]. 遗传学报, 2003(2):183-188.
Guo WZ, Wang K, Zhang TZ.A and D genome evolution in gossypium revealed using SSR molecular markers[J]. Journal of Genetics and Genomics, 2003(2):183-188.
[5] 蔡小彦, 王玉红, 许艳超, 等. 棉花种间远缘杂交及F₁真假杂种的SSR鉴定方法研究[J]. 中国棉花, 2018, 45(11):6-9, 44.
Cai XY, Wang YH, Xu YC, et al.Identification of cotton interspecific hybrids with SSR markers[J]. China Cotton, 2018, 45(11):6-9, 44.
[6] 冯文林, 陈全家, 曲延英. 棉花海陆棉杂交后代农艺与纤维品质性状相关性研究[J]. 新疆农业科学, 2015, 52(3):393-401.
Feng WL, Chen QJ, Qu YY.Relativity of agronomic traits and fiber quality of sea-land cotton hybrid progeny[J]. Xinjiang Agricultural Sciences, 2015, 52(3):393-401.
[7] 申状状, 李昱樱, 荣二花, 等. 陆地棉和野生斯特提棉种间异源六倍体的合成与性状鉴定[J]. 作物学报, 2019, 45(4):628-634.
Shen ZZ, Li YY, Rong EH, et al.Allohexaploid synthesis and its characteristic identification between cotton species Gossypium hirsutum and G. sturtianum[J]. Acta Agronomica Sinica, 2019,45(4):628-634.
[8] Konan NO, Mergeai G.Relationship between meiotic behaviour and fertility in backcross-1 derivatives of the[(Gossypium hirsutum × G. thurberi)2 × G. longicalyx]trispecies hybrid[J]. Comparative Cytogenetics, 2020, 14(1):75-95.
[9] 申状状, 杨娜, 荣二花, 等. 亚洲棉与瑟伯氏棉种间远缘杂种的合成及其性状鉴定[J]. 分子植物育种, 2017, 15(6):2291-2297.
Shen ZZ, Yang N, Rong EH, et al.Synthesis and identification of interspecific hybrid between Gossypium arboreum L. and G. thurberi T.[J]. Molecular Plant Breeding, 2017, 15(6):2291-2297.
[10] 崔淑芳, 钱玉源, 王广恩, 等. 远缘杂交种冀棉25的选育及其育种利用[J]. 中国棉花, 2018, 45(3):21-25.
Cui SF, Qian YY, Wang GE, et al.Breeding and application of intergeneric hybridization cotton variety Jimian 25[J]. China Cotton, 2018, 45(3):21-25.
[11] 肖松华, 刘剑光, 赵君, 等. 棉花远缘杂交创制抗黄萎病新种质[J]. 棉花学报, 2015, 27(6):524-533.
Xiao SH, Liu JG, Zhao J, et al.Creation of a new resistant germplasm to verticillium wilt by distant hybridization in upland cotton[J]. Cotton Science, 2015, 27(6):524-533.
[12] 郭宝生, 韩泽林, 耿军义, 等. 陆、海、瑟棉花远缘杂交后代的遗传改良[J]. 华北农学报, 2007(S2):85-88.
Guo BS, Han ZL, Geng JY, et al.Genetic improvement of distant hybridization offsprings come from G.hirsutum and G.barbadense and G.thurberi[J]. Acta Agriculturae Boreali-Sinica, 2007(S2):85-88.
[13] 陈煜. 棉属(Gossypium)种间杂交研究及陆地棉(G. hirsutum)-澳洲棉(G. australe)异附加系的培育[D]. 南京:南京农业大学, 2014.
Chen Y.Study on the interspecific hybridization in Gossypium and development of alien addition lines from Gossypium australe to Gossypium hirsutum[D]. Nanjing: Nanjing Agricultural University, 2014.
[14] 吕昊敏, 周仁超, 施苏华. 生态物种形成及其研究进展[J]. 生物多样性, 2015, 23(3):398-407.
Lv HM, Zhou RC, Shi SH.Recent advances in the study of ecological speciation[J]. Biodiversity Science, 2015, 23(3):398-407.
[15] 吕锐玲, 周强, 王欢, 等. 植物远缘杂交中生殖隔离研究进展[J]. 湖北农业科学, 2016, 55(24):6337-6341.
Lv RL, Zhou Q, Wang H, et al.Research progress of reproductive isolation in distant hybridization of plants[J]. Hubei Agricultural Sciences, 2016, 55(24):6337-6341.
[16] 王志龙, 姜茹琴, 何鉴星, 等. 棉属海岛棉×拟似棉F₁不育性研究[J]. 遗传学报, 1997(4):368-372.
Wang ZL, Jiang RQ, He JX, et al.Study on sterility of hybrid between G. barbadense and G. gossypioides[J]. Journal of Genetics and Genomics, 1997(4):368-372.
[17] Kwiatek MT, Kurasiak-Popowska D, Mikolajczyk S, et al.Cytological markers used for identification and transfer of Aegilops spp. chromatin carrying valuable genes into cultivated forms of Triticum[J]. Comparative Cytogenetics, 2019, 13(1):41-59.
[18] 管敏. SSR分子标记在棉花遗传育种中的应用分析[J]. 生物技术世界, 2014(10):25.
Guan M.Analysis on the application of SSR molecular markers in cotton genetics and breeding[J]. Biotechworld, 2014(10):25.
[19] 付小琼, 叶武威. 亲本未知的棉花F₁杂种及其F₂单株的SSR分子鉴定[J]. 分子植物育种, 2012, 10(5):600-606.
Fu XQ, Ye WW.Identification of F1 hybrid and F2 individual in cotton with unknown parents[J]. Mol Plant Breed, 2012, 10(5):600-606.
[20] 石建斌, 周红, 王宁, 等. 棉花SSR标记种质资源纯度鉴定及遗传多样性分析[J]. 生物技术通报, 2018, 34(7):138-146.
Shi JB, Zhou H, Wang N, et al.Purity identification and genetic diversity analysis of cotton germplasm resources using SSR markers[J]. Biotechnology Bulletin, 2018, 34(7):138-146.
[21] 薛玉前, 庄木, 方智远, 等. 植物杂种致死研究进展[J]. 中国农业科技导报, 2015, 17(2):1-7.
Xue YQ, Zhuang M, Fang ZY, et al.Research progress in plant hybrid lethality[J]. Journal of Agricultural Science and Technology, 2015,17(2):1-7.
[22] 戴宝生. 棉花海陆种间偏分离与杂种衰败遗传机理及效应研究[D]. 武汉:华中农业大学, 2016.
Dai BS.The phenotypic effection and genetic mechanism of segreation distortion and hybrid breakdown in the interspecific poplations of G.Hirsutum L and G.Barbadense L[D]. Wuhan: Huazhong Agricultural University, 2016.
[23] Phillips LL, Reid RK.Interspecific incompatibility in Gosspium:II. light and electron microscopic studies of cell necrosis and tumorigenesis in hybrid of G. klotzschianum[J]. American Journal of Botany, 1975, 62:790-796.
[24] Wendel JF, Cronn RC.Polyploidy and the evolutionary history of cotton[J]. Advances in Agronomy, 2003, 78:129-186.
[25] Li F, Fan G, Lu C.et al.Genome sequence of cultivated upland cotton(Gossypium hirsutum TM-1)provides insights into genome evolution[J]. Nat Biotechnol, 2015, 33:524-530.
[26] Wang K, Wang Z, Li F, et al.The draft genome of a diploid cotton Gossypium raimondii[J]. Nature Genetics, 2012, 44(10):1098-1103.
[27] 梅磊, 李玲, 肖钦之, 等. 陆地棉植物络合素合酶基因的鉴定与功能预测[J]. 棉花学报, 2018, 30(3):215-223.
Mei L, Li L, Xiao QZ, et al.Genome-wide identification and functional prediction of phytochelatin synthase gene in upland cotton[J]. Cotton Science, 2018, 30(3):215-223.
[28] 吴玉香. 棉属遗传多样性与栽培棉种间杂交研究[D]. 杭州:浙江大学, 2007.
Wu YX.Study on genetic diversity of gossypium and interspecific hybridization of cultivated cotton[D]. Hangzhou:Zhejiang University, 2007.