Liu Chuanxiang, Yuan Daojun, Zhang Xianlong, Lin Zhongxu
National Key Laboratory of Crop Genetic Improvement and National Centre of Plant Gene Research, Huazhong Agricultural University, Wuhan 430070, Hubei, People's Republic of China.
J Genet. 2013;92(2):175-81.
Gossypium hirsutum and G. barbadense are two cultivated tetraploid cotton species with differences in fibre quality. The fibre of G. barbadense is longer, stronger and finer than that of G. hirsutum. To isolate genes expressed differently between the two species during fibre development, cDNA-SRAP (sequence-related amplified polymorphism) was applied. This technique was used to analyse genes at different stages of fibre development in G. hirsutum cv. Emian22 and G. barbadense acc. 3-79, the parents of our interspecific mapping population. A total of 4096 SRAP primer combinations were used to screen polymorphism between the DNA of the parents, and 275 highly polymorphic primers were picked out to analyse DNA and RNA from leaves and fibres at different developmental stages of the parents. A total of 168 DNA fragments were isolated from gels and sequenced: 54, 30, 38 and 41 from fibres of 5, 10, 15 and 20 days post-anthesis, respectively, and five from multi stages. To genetically map these sequences, 104 sequence-specific primers were developed and were used to screened polymorphism between the mapping parents. Finally, six markers were mapped on six chromosomes of our backbone interspecific genetic map. This work can give us a primary knowledge of differences in mechanism of fibre development between G. hirsutum and G. barbadense.
陆地棉和海岛棉是两个栽培四倍体棉花物种，在纤维品质上存在差异。海岛棉的纤维比陆地棉更长、更强且更细。为了分离这两个物种在纤维发育过程中差异表达的基因，应用了cDNA-SRAP（序列相关扩增多态性）技术。该技术用于分析陆地棉品种鄂棉22和海岛棉品系3-79（我们种间作图群体的亲本）纤维发育不同阶段的基因。总共使用了4096对SRAP引物组合来筛选亲本DNA之间的多态性，并挑选出275对高度多态性引物来分析亲本不同发育阶段叶片和纤维的DNA和RNA。从凝胶中分离出总共168个DNA片段并进行测序：分别从开花后5天、10天、15天和20天的纤维中分离出54个、30个、38个和41个，从多个阶段分离出5个。为了对这些序列进行遗传定位，开发了104个序列特异性引物，并用于筛选作图亲本之间的多态性。最后，6个标记被定位在我们的骨干种间遗传图谱的6条染色体上。这项工作可以让我们初步了解陆地棉和海岛棉纤维发育机制的差异。
