Yasumoto Keita, Terachi Toru, Yamagishi Hiroshi
Department of Biotechnology, Faculty of Engineering, Kyoto Sangyo University, Motoyama, Kamigamo, Kita-ku, Kyoto 603-8555, Japan.
Genome. 2009 Jun;52(6):495-504. doi: 10.1139/g09-026.
To reveal the molecular and genetic mechanism of fertility restoration in Ogura male sterility in Japanese wild radish (Raphanus sativus var. hortensis f. raphanistroides), we investigated fertility restoration of a plant that lacks the dominant type of orf687, a previously identified fertility restorer gene. A total of 100 F2 plants were made from the cross between a male-sterile strain with the Ogura cytoplasm, 'MS-Gensuke', and a Japanese wild radish plant. Segregation of pollen fertility in the F2 plants led us to assume that 2 dominant complementary genes controlled the fertility restoration of the plants. However, the fertility of 27 of 59 male-fertile plants was not completely restored, resulting in a group of plants with partial male fertility. Northern blot analysis of the CMS-associated gene orf138 indicated that one restorer allele (termed Rft) was involved in the processing of orf138 RNA. Rapid amplification of cDNA ends (RACE) and subsequent Northern blot analysis confirmed that the orf138 transcript lost a 5' part of the coding region of the orf138 gene in the restored plants. The accumulation of ORF138 protein was significantly reduced by Rft, but trace amounts of the protein were recognized in both partially male-fertile and male-sterile plants with Rft. The relationship of pollen fertility and segregation of co-dominant sequence tagged site (STS) markers in the F2 generation suggested that the penetrance of Rft was so low that Rft needs suitable conditions to function sufficiently for the complete restoration of fertility.
为揭示日本野生萝卜(Raphanus sativus var. hortensis f. raphanistroides)中萝卜细胞质雄性不育育性恢复的分子和遗传机制,我们研究了一株缺乏之前鉴定出的育性恢复基因orf687显性类型的植株的育性恢复情况。用具有萝卜细胞质的雄性不育系‘MS - Gensuke’与一株日本野生萝卜杂交,共获得了100株F2植株。F2植株花粉育性的分离情况使我们推测有2个显性互补基因控制着植株的育性恢复。然而,59株雄性可育植株中有27株的育性未完全恢复,形成了一组具有部分雄性育性的植株。对CMS相关基因orf138的Northern杂交分析表明,一个恢复基因等位基因(命名为Rft)参与了orf138 RNA的加工过程。cDNA末端快速扩增(RACE)及随后的Northern杂交分析证实,在育性恢复的植株中,orf138转录本缺失了orf138基因编码区的5′部分。Rft显著降低了ORF138蛋白的积累,但在具有Rft的部分雄性可育和雄性不育植株中都检测到了微量的该蛋白。F2代中花粉育性与共显性序列标签位点(STS)标记分离的关系表明,Rft的外显率很低,以至于Rft需要合适的条件才能充分发挥作用以实现育性的完全恢复。