陆地棉与野生斯特提棉远缘杂种性状鉴定及遗传解析

doi:10.13560/j.cnki.biotech.bull.1985.2020-1359

摘要/Abstract

摘要：

棉花远缘杂交是种质创新和新品种选育的重要途径。以实验室前期合成的陆地棉与野生斯特提棉远缘杂种为材料,对其进行形态学、细胞遗传学及SSR分子标记鉴定。形态学分别从植株、叶片和花三方面进行了双亲和杂种的分析比较,结果表明杂种植株高于双亲,叶片大于双亲,表现出显著的杂种优势;叶色与父本相近,为翠绿色,杂种花基部花斑与父本相近,为深红色;细胞遗传学上对杂种进行了花粉母细胞减数分裂观察,结果表明不育杂种F₁在减数分裂过程中出现许多异常行为,其中二分体占24.00%,三分体占10.40%,四分体占40.60%,多分体占25.00%。在四分体中,正常四分体占72.91%,异常占27.09%,以至于最后形成很多异常花粉粒,所占比例为24.00%,正常花粉粒占76.00%,这是杂种不育的主要原因;最后通过SSR分子标记鉴定结果表明,杂种F1不仅扩增出双亲的互补带,还扩增出双亲没有的特异性条带,遗传成分比例分别为：父本占7.69%,母本占34.62%,双亲的互补带占23.07%,特异性新带占34.62%,既表明在杂交过程中发生了基因重组,也从分子水平证实了该杂种是陆地棉和斯特提棉的真杂种,同时为棉花遗传育种和种质创新提供了有价值的材料。

关键词: 斯特提棉, 远缘杂交, 性状鉴定, SSR

Abstract:

Distant hybridization of cotton is an important approach for germplasm innovation and breeding of new varieties. In this study,the distant hybrid of Gossypium hirsutum and Gossypium sturtianum synthesized in the laboratory were used as materials and was identified in terms of morphology,cytogenetics and SSR molecular markers. Morphological analysis and comparison of the parents and hybrids were carried out from three aspects of plants,leaves and flowers respectively. Results showed that the hybrid plants were higher than that of the parents and the leaves were larger than that of the parents,demonstrating obvious heterosis. The colors of the leaves were emerald green,similar to that of the male parent. And the colors of the basal spots of hybrids were dark red,similar to that of the male parent. As for cytogenetics,pollen mother cell meiosis was observed in hybrids,and the results showed that there were many abnormal behaviors in the process of meiosis in sterile hybrid F1,among which diad,triad,tetrad and polyad accounted for 24.00%,10.40%,40.60% and 25.00%,respectively. In the tetrad,72.91% of normal tetrad and 27.09% of abnormal tetrad resulted in the formation of 24.00% abnormal pollen grains and 76.00% of normal pollen grains,which were the main reasons for hybrid sterility. Finally,the identification of SSR molecular marker showed that F1 hybrids not only amplified the complementary bands of the parents,but also amplified the specific bands that the parents did not have. The proportions of genetic components were as follows：7.69% for the male parent,34.62% for the female parent,23.07% for the parental complementary bands,and 34.62% for specific new bands. It is not only suggested that the gene recombination occurred in the hybridization process,but also confirmed that the hybrid was the true hybrid of G. hirsutum and G. sturtianum at the molecular level. At the same time,this study provides valuable materials for the genetic breeding and germplasm innovation of cotton.

Key words: Gossypium sturtianum, distant hybridization, character identification, SSR

赵祝跃, 申状状, 王一帆, 杨亚杰, 荣二花, 吴玉香. 陆地棉与野生斯特提棉远缘杂种性状鉴定及遗传解析[J]. 生物技术通报, 2021, 37(5): 19-27.

ZHAO Zhu-yue, SHEN Zhuang-zhuang, WANG Yi-fan, YANG Ya-jie, RONG Er-hua, WU Yu-xiang. Character Identification and Genetic Analysis of Distant Hybrid Between Gossypium hirsutum and Gossypium sturtianum[J]. Biotechnology Bulletin, 2021, 37(5): 19-27.

图/表 9

表1 供试材料及其地理分布

Table 1 Materials and their geographical distribution

材料Materials	染色体组Genome	分布地Distribution
陆地棉G. hirsutum	AD₁	世界栽培种 World cultivated species
斯特提棉G. sturtianum	C₂	澳大利亚中部南部 Central and south Australia

图1 亲本与杂种的植株比较 A：母本陆地棉;B：杂种F1;C：父本斯特提棉

Fig.1 Plant comparison between parents and hybrid A: G. hirsutum. B: Hybrid F1. C: G. sturtianum

图2 亲本与杂种的叶片比较（标尺：20 mm） A：母本陆地棉;B：父本斯特提棉;C：杂种F1

Fig.2 Leaf comparison between parents and hybrid (Ruler: 20 mm) A: G. hirsutum. B: G. sturtianum. C: Hybrid F1

图3 亲本与杂种的花朵比较 A：母本陆地棉;B：杂种F1;C：父本斯特提棉

Fig.3 Flower comparison between parents and hybrid A: G. hirsutum. B: Hybrid F1. C: G. sturtianum

图4 杂种的正常和异常减数分裂行为（标尺：20 μm） A：异常二分体;B：三分体;C：异常四分体;D：正常四分体;E-H：多分体

Fig.4 Normal and abnormal meiosis behavior of G. hirsutum×G. sturtianum F1 hybrid (Ruler: 20 μm) A: Abnormal dyad. B: Triad. C: Abnormal tetrad. D: Normal tetrad. E-H: Polyad

表2 杂种减数第二次分裂末期的多分体数量及比例

Table 2 Number of multispores in telophase II of ADC hybrid meiosis

形式Shape	二分体Dyad	三分体Triad	四分体Tetrad	多分体Polyad	总数Total
No	120	52	203	125	500
%	24.00	10.40	40.60	25.00	100

图5 杂种减数分裂后的正常和异常花粉粒（标尺：20 μm） A：正常花粉粒;B：一个正常和一个发育不全的花粉粒;C：一个正常和两个未发育完全的花粉粒;D：畸形花粉粒;E：一个正常和一个未发育完全花粉粒;F：破裂的花粉粒;G：三个正常的花粉粒;H：一个正常和一个未发育完全花粉粒

Fig.5 Various normal and abnormal pollen grains after meiosis of F1 hybrid (Ruler: 20 μm) A: Normal pollen grain. B: One normal and one underdeveloped pollen grain. C: One normal and two underdeveloped pollen grains. D: A malformed pollen grain. E: One normal and one underdeveloped pollen grain. F: A cracked pollen grain. G: Three normal pollen grains. H: One normal and one underdeveloped pollen grain

图6 SSR引物在杂种及父母本中的扩增结果

Fig.6 Amplification results of SSR primers in hybrid and parents

表3 杂种SSR的多态性来源分布

Table 3 Source distribution of SSR polymorphisms in hybrid

材料Material	杂种Hybrid	母本Female parent	父本Male parent	双亲互补带Parental complementary band	特异性条带Specific band
扩增条带 Amplification bands	26	9	2	6	9
遗传比例 Genetic ratio/%	-	34.62	7.69	23.07	34.62

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