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首次将PMC-FISH(花粉母细胞荧光原位杂交)技术应用于四倍体栽培棉与二倍体野生棉杂交所形成的三倍体杂种棉F1中,成功的鉴定了目标染色体中的单价体、双价体、多价体,还发现在非目标染色体上有星状和片段状的杂交信号。在以A组棉基因组DNA作为探针的PMC-FISH中,分别对三倍体杂种棉F1及其母本四倍体栽培棉的减数分裂中期I的染色体构型进行了鉴定。结果显示,四个三倍体杂种棉F1的减数分裂时期染色体的构型分别是:陆地棉(AD)1×雷蒙德氏棉(D5)为1IIIaad+1IIaa+4IIad+7IIdd+5Ⅰa+7Ⅰd;海岛棉(AD)2×旱地棉(D4)为1Ⅴaaaad+1IIIadd+2IIad+8IIdd+6Ⅰa+5Ⅰd;陆地棉(AD)1×澳洲棉(C3)为2IIIadd(adc or acc)+1IIaa+9Ⅰa+4IIcc(dc or dd)+14Ⅰd(c);陆地棉(AD)1×南岱华棉(C1-n)为:1IIaa+1IIad(ac)+10Ⅰa+6IIcc(dc or dd)+13Ⅰd(c)。然而,两个四倍体棉染色体的构型都是13IIaa+13IIdd。上述结果还显示,AD×D的两个杂种组合中,二价体多,单价体少,AD×C的两个杂种组合中,二价体少,单价体多;并且AD×D型杂种中A亚组染色体单体的数目比其在AD×C型杂种中的更少。这说明,与C染色体组相比较,D染色体组与四倍体棉种的亲缘关系更近。同时,我们的结果也证明PMC-FISH技术在分析三倍体杂种棉F1的亲缘关系中起着不可取代的作用。
The first application of PMC-FISH (pollen in situ hybridization) technique to triploid hybrid cotton F1 formed by crossing tetraploid cotton with diploid wild cotton successfully identified monovalent , Bivalents, polyvalents, also found in non-target chromosomes and star-shaped cross-like hybridization signal. The chromosome configuration of metaphase I in triploid hybrid cotton F1 and its female tetraploid cotton was identified in PMC-FISH using A group of cotton genomic DNA as probe. The results showed that the chromosome configurations at the meiosis stage of four triploid F1 hybrids were: 1IIIaad + 1IIaa + 4IIad + 7IIdd + 5Ⅰa + 7Ⅰd (AD) 2 × dry cotton (D4) were 1 ± aaaaad + 1IIIadd + 2IIad + 8IIdd + 6Ⅰa + 5Ⅰd; 1 × Australia cotton (C3) was 2IIIadd (adc or acc) + 1IIaa + 9Ⅰa + 4IIcc (dc or dd) + 14Id (c); cotton (AD) 1 × Nan Daihua cotton (C1-n): 1IIaa + 1IIad (ac) + 10Ⅰa + 6IIcc (dc or dd) . However, the configuration of both tetraploid cotton chromosomes is 13IIaa + 13IIdd. The above results also show that the AD × D two hybrid combinations, more bivalent, less monovalent, AD × C two hybrid combinations, fewer bivalent, monovalent more; and AD × D-type hybrids The number of chromosomal monomers in the A subgroup is less than that in the AD x C type hybrids. This shows that, compared with the C genome, D genome and tetraploid cotton closer genetic relationship. At the same time, our results also prove that PMC-FISH technology plays an irreplaceable role in the analysis of the genetic relationship of triploid hybrid cotton F1.