生物技术通报 ›› 2022, Vol. 38 ›› Issue (5): 64-73.doi: 10.13560/j.cnki.biotech.bull.1985.2021-1288

• 研究报告 • 上一篇    下一篇

草棉同源多倍体后代筛选及性状鉴定

杨亚杰(), 李昱樱, 申状状, 陈天, 荣二花, 吴玉香()   

  1. 山西农业大学农学院,太谷 030801
  • 收稿日期:2021-10-10 出版日期:2022-05-26 发布日期:2022-06-10
  • 作者简介:杨亚杰,女,硕士研究生,研究方向:种质创新与遗传工程;E-mail: 2876506015@qq.com
  • 基金资助:
    国家自然科学基金项目(31171599);山西省基础研究计划项目(自由探索类)(20210302124154)

Selection and Character Identification for Autopolyploid Progenies of Gossypium herbaceum

YANG Ya-jie(), LI Yu-ying, SHEN Zhuang-zhuang, CHEN Tian, RONG Er-hua, WU Yu-xiang()   

  1. College of Agriculture,Shanxi Agricultural University,Taigu 030801
  • Received:2021-10-10 Published:2022-05-26 Online:2022-06-10

摘要:

多倍化是物种形成及植物育种的重要途径。以实验室前期合成的草棉同源多倍体后代S1为材料,对其进行流式细胞术、形态学、细胞学和SRAP分子标记鉴定。流式细胞倍性鉴定表明,以二倍体的荧光峰值作为参照,8株后代中有5株三倍体和3株四倍体。形态学鉴定表明,二倍体表现为株高、茎细、叶薄、叶片较小、叶色翠绿、花朵较小,三倍体和四倍体则表现为植株矮小、茎秆粗壮、叶片增厚和皱缩、叶片较大、叶色浓绿、花朵较大。细胞学鉴定表明,随着倍性的增加,气孔密度呈下降趋势,而气孔长度、叶绿体数和花粉粒直径均呈上升趋势;二、三和四倍体草棉在花粉母细胞减数分裂过程中均出现正常和异常行为,包括单分体(0.17%,2.50%和2.17%)、二分体(0.33%,0.67%和0)、三分体(4.17%,10.00%和2.33%)、正常四分体(94.83%,54.00%和73.67%)、异常四分体(0.17%,0.33%和0.83%)和多分体(0.33%,32.50%和21.00%);异常多分体均不能形成正常花粉粒,所以三者的正常花粉粒所占比例分别为95.33%、58.83%和77.67%。SRAP分子标记鉴定表明,三倍体和四倍体不仅扩增出与二倍体完全一致的条带,还出现了条带的缺失与新的特异性条带,其中三倍体的遗传比例分别为80.81%、6.73%和12.46%,四倍体的遗传比例分别为76.59%、10.37%和13.04%,从分子水平证明了多倍体的真实性。

关键词: 草棉, 同源多倍体, 性状鉴定, 分子标记

Abstract:

Polyploidization is an important way of speciation and plant breeding. In this study,the autopolyploid of Gossypium herbaceum progenies S1 in the early stage were used as materials,and flow cytometry,morphological,cytological and SRAP molecular marker were used to identify them. The results of flow cytometry for ploidy identification indicated that the fluorescence peak value of diploids was used as a reference,among eight polyploid progenies,there were 5 triploid plants and 3 tetraploid plants,respectively. The results of morphological identification showed that compared with polyploids,diploid plants were taller with thinner stalks,as well as thinner,smaller and emerald green leaves,and flowers were smaller. Compared with diploids,triploids and tetraploids all presented shorter and thicker stalks,as well as thicker,shrunken,larger and dark green leaves,and flowers were larger. The results of cytological identification demonstrated that with the increase of ploidy level,it showed a downward trend for stomatal density,while it all showed an upward trend for the length of stomatal,the number of chloroplasts and the diameter of pollen grains. There were normal and abnormal behaviors during meiosis in pollen mother cell for all diploids,triploids and tetraploids,including monad(0.17%,2.50% and 2.17%),dyad(0.33%,0.67% and 0),triad(4.17%,10.00% and 2.33%),normal tetrad(94.83%,54.00% and 73.67%),abnormal tetrad(0.17%,0.33% and 0.83%)and polyad(0.33%,32.50% and 21.00%)respectively. The abnormal polyad could not produce normal pollen grains,thus the proportion of normal pollen grains in the diploids,triploids and tetraploids were 95.33%,58.83% and 77.67% respectively. The results of SRAP molecular marker identification revealed that there were three polymorphism bands amplified in triploids and tetraploids:including completely consistent with diploids bands,lost bands and new specific bands when comparing with diploids. The genetic proportion of three band types in the triploids were 80.81%,6.73% and 12.46%,and that of the tetraploids were 76.59%,10.37% and 13.04%,respectively. This confirmed the authenticity of polyploidy at the molecular level.

Key words: Gossypium herbaceum, autopolyploid, character identification, molecular marker