橡胶草和枣红蒲公英杂交后代SSR分子鉴定及性状分析

SSR molecular identification and trait analysis of hybrid progeny between Taraxacum kok-saghyz Rodin and Taraxacum spadiceum

  • 摘要: 【目的】 开展自交不亲和橡胶草与无融合生殖枣红蒲公英的杂交试验,以期通过种间杂交产生无融合生殖的橡胶草,从而克服橡胶草因自交不亲和而无法产生基因型稳定种子的问题,为橡胶草遗传学研究及商业化品种培育提供理论参考。【方法】 以无融合生殖的三倍体枣红蒲公英作为父本、自交不亲和的二倍体橡胶草作为母本,采用人工授粉的方式获得杂交子代。设计并筛选出稳定且具有特异性的SSR分子标记引物,对子代进行杂交真实性鉴定,并对鉴定出的杂种F1代进行生殖方式鉴定、表型观察、根胶含量初步观测及倍性分析。【结果】 杂交试验共对3759朵花进行授粉,收获157个至少有1个饱满种子的种子球,最终萌发得到203株健康后代,结实率约为1‰,杂交成功率仅为0.03‰。共筛选出3对条带清晰且重复性好的特异性引物SSR48、SSR49和SSR51,其中SSR48、SSR49为共显性引物,能同时扩增出父、母本的特异性条带;SSR51只能扩增出父本特异性条带,3对SSR引物用于枣红蒲公英和橡胶草的真杂交子代鉴定。SSR分子标记检测结果显示,203株子代中有197株仅有母本特异性条带,为橡胶草自交后代;其余6株既有母本特异性条带又有父本特异性条带,为真杂交子代,真杂种率约为3.0%。杂种F1代中有5株真杂种植株的生殖方式为无融合生殖,均为三倍体,其叶片形状、大小和花莛长度更接近父本,几乎不产天然橡胶。【结论】 二倍体橡胶草与三倍体枣红蒲公英通过人工授粉可产生可育的杂交后代,但杂交效率极低。育成的橡胶草自交子代和少量无融合生殖杂交后代可用于后续橡胶草育种研究。

     

    Abstract: 【Objective】 This study aimed to conduct hybridization experiments between self-incompatible Taraxacum kok-saghyz Rodin (Tk) and apomictic Taraxacum spadiceum (Ts) to generate apomictic Tk through interspecific hybri-dization for overcoming the problem of inability of Tk to produce seeds with stable genotype due to self-incompatibility, so as to provide a theoretical reference for genetic research and commercial cultivar development in Tk. 【Method】 The apomictic triploid Ts was used as the male parent line, and the self-incompatible diploid Tk as the female parent line. Hybrid progeny were obtained through artificial pollination. Identification of reproductive mode, phenotypic observation, preliminary measurement of root rubber content,and ploidy analysis were conducted for the identified F1 hybrid. 【Result】 A total of 3759 flowers were pollinated, yielding 157 seed heads, each containing at least one plump seed. Ultimately, 203 healthy progeny germinated, with the seed setting rate was about 1‰, and the hybridization success rate was only 0.03‰. Three pairs of specific primers (SSR48, SSR49, and SSR51) with clear bands and good reproducibility were screened, and SSR48 and SSR49 were codominant primers, amplifying specific bands from male and female parent lines simultaneously. SSR51 could be used to amplify a male-parent-specific band; and three SSR primer pairs were used to identify true hybrids between Ts and Tk. SSR marker detection results showed that among the 203 progeny, 197 exhibited only the maternal specific band, identified as selfed progeny of Tk. The rest six plants possessed both maternal and paternal specific bands, confirming them as the F1 true hybrid, resulting in a true hybrid rate around 3.0%. Among the F1 hybrid, five were true hybrid plants exhibiting apomictic reproduction and were triploid. Their leaf shape, size, and scape length more closely resembled the male parent line, and they produced almost no natural rubber. 【Conclusion】 Fertile hybrid progeny can be generated between diploid Tk and triploid Ts through artificial pollination, albeit with extremely low efficiency. The selfed Tk progeny and the small number of apomictic hybrids obtained in this study can be used for subsequent Tk breeding research.

     

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