棉属AADD异源四倍体后代筛选及性状鉴定  

Screening and characterization of AADD allotetraploid in genus Gossypium

作　　者：陈天 李昱樱 杨亚杰 候林慧 常永春 荣二花[1] 吴玉香[1] Chen Tian;Li Yuying;Yang Yajie;Hou Linhui;Chang Yongchun;Rong Erhua;Wu Yuxiang(College of Agronomy,Shanxi Agricultural University,Jinzhong 030801,China)

机构地区：[1]山西农业大学农学院,山西晋中030801

出　　处：《山西农业大学学报（自然科学版）》2022年第6期72-80,共9页Journal of Shanxi Agricultural University(Natural Science Edition)

基　　金：山西省基础研究计划项目(自由探索类)(20210302124154);国家自然科学基金(31171599)。

摘　　要：[目的]棉属异源四倍体棉种的起源是棉属进化研究的热点,本研究模拟自然界异源四倍体棉种形成过程,以草棉作母本,野生雷蒙德氏棉作父本进行远缘杂交,获得杂种F1,然后采用秋水仙碱对杂种进行染色体加倍,合成人工异源四倍体,旨在探究人工异源四倍体物种形成后的遗传规律。[方法]进行形态性状鉴定和流式细胞检测,以及细胞遗传学和分子水平鉴定。[结果]形态性状分析表明:多倍体植株叶片形状与母本相似,叶片大小介于两亲本之间。流式细胞检测结果表明:对照陆地棉的荧光峰值为215.94,所选5株多倍体植株荧光峰值在195.52与258.46之间,与陆地棉接近。细胞遗传学鉴定表明:人工异源四倍体S2代正常四分体比例为23.4%,S3代正常四分体比例为44.8%;S2代正常花粉粒比例为55.4%,S3代正常花粉粒比例为64.4%。SRAP分子标记分析说明:人工异源四倍体不仅扩增出与陆地棉和草棉相同的条带,还扩增出特异性条带,其遗传比例分别为46%、31%和23%。[结论]本研究结果证明了人工合成多倍体的真实性以及多倍体育性在逐代恢复,并且表明多倍体在分子水平上出现了染色体重排,和陆地棉具有一定的同源性。[Objective]The origin of allotetraploid species in Gossypium is a hot topic in Gossypium evolution research. In the present study,we simulated the formation process of tetraploid cotton species in nature using Gossypium arboreum as the maternal parent and Gossypium raimondii as the paternal parent in crossbreeding to obtain the hybrid F1. Then, colchicine was used to double the chromosomes of the hybrid to synthesize an artificial tetraploid species in order to explore the genetic laws of the species after its formation.[Methods]Morphological trait identification, flow cytometry, cytogenetics, and molecular analyses were carried out to explore the genetic regular pattern of allotetraploid speciation.[Results]The morphological character analysis showed that the leaf shape of the polyploid plant was similar to that of G. herbaceun, and the leaf size was between its parents of G. herbaceun and G. raimondii. Flow cytometric analysis results showed that the fluorescence peak value of G. hirsutum was 215. 94, and the peak values of five selected polyploid plants were between 195. 52 and 258. 46, which were similar to that of G. hirsutum. Cell genetic identification indicated that the proportion of normal tetraploid cells in the S2 generation of the artificial allotetraploid was 23. 4%, and in the S3 generation was 44. 8%;the proportion of normal pollen grains in the S2 generation was 55. 4%, and in the S3 generation was 64. 4%. Sequence-related amplified polymorphism(SRAP)molecular marker analysis indicated that the artificial allotetraploid not only amplified bands that were the same as those of G. hirsutum and G.arboreum, but also amplified specific bands, with a genetic proportion of 46%, 31%, and 23%.[Conclusion]The results of this study confirmed the authenticity of artificially synthesized polyploidy and the recovery of polyploidy in successive generations, and have also shown that there is chromosomal reorganization at the molecular level in polyploids, and a certain degree of homology with G. hirsutum.

关 键 词：人工异源四倍体 性状鉴定 减数分裂 流式细胞 SRAP分子标记 

分 类 号：S335.1[农业科学—作物遗传育种]