高通量转录组测序技术在植物雄性不育研究中的应用

植物雄性不育是指植物雄蕊发育受阻不能产生正常有功能花粉的现象。植物雄性不育不仅是生殖生理研究的宝贵材料,也是植物杂种优势利用的重要工具。由于高通量转录组测序技术几乎可以检测细胞内所有mRNA及非编码RNA的信息,已被广泛应用于生命科学研究的各项领域。在植物雄性不育相关研究中,高通量转录组测序技术在不同物种、不同败育类型中的应用已有报道,这为研究者在转录组水平综合了解植物雄性不育的分子机制及代谢网络提供了帮助。本文从测序文库构建策略、差异表达基因、非编码RNA的功能特征等方面综述了高通量转录组测序在植物雄性不育机理方面的研究进展,并探讨了转录组测序技术在花粉败育机制解析及育性相关基因定位中的应用价值,以期为植物雄性不育的相关研究提供参考。     

Fine mapping of the recessive genic male sterility gene (<Emphasis Type="Italic">Bnms3</Emphasis>) in <Emphasis Type="Italic">Brassica napus</Emphasis> L.

The Brassica napus oilseed rape line, 7-7365AB, is a recessive epistatic genic male sterile (RGMS) two-type line system. The sterility is controlled by two pairs of recessive duplicate genes (Bnms3 and Bnms4) and one pair of recessive epistatic inhibitor gene (Bnrf). Homozygosity at the Bnrf locus (Bnrfrf) inhibits the expression of the two recessive male sterility genes in homozygous Bnms3ms3ms4ms4 plants and produces a male fertile phenotype. This line has a good potential for heterosis utilization but it is difficult to breed heterotic hybrids without molecular markers. To develop markers linked to the BnMs3 gene, amplified fragment length polymorphism (AFLP) technology was applied to screen the bulks of sterile and fertile individuals selected randomly from a population of near-isogenic lines (NIL) consisting of 2,000 plants. From a survey of 1,024 primer combinations, we identified 17 AFLP markers linked to the BnMs3 gene. By integrating the previous markers linked to the BnMs3 gene into the genetic map of the NIL population, two markers, EA01MC12 and EA09P06, were located on either side of the BnMs3 gene at a distance of 0.1 and 0.3 cM, respectively. In order to use the markers for male sterile line breeding, five AFLP markers, P05MG05, P03MG04, P11MG02, P05MC11₂₅₀, and EA09P06, were successfully converted into sequence characterized amplified region (SCAR) markers. Two of these, P06MG04 and sR12384, were subsequently mapped on to linkage group N19 using two doubled-haploid mapping populations available at our laboratory derived from the crosses Tapidor × Ningyou7 and Quantum × No2127-17. The markers found in the present study should improve our knowledge of recessive genic male sterility (RGMS), and accelerate the development of male sterile line breeding and map-based cloning.