National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, 430070, China.
Mol Biol Rep. 2011 Jan;38(1):1-8. doi: 10.1007/s11033-010-0070-x. Epub 2010 Mar 21.
'Zigui shatian' pummelo (Citrus grandis Osbeck) is one nature mutant from 'Shatian' pummelo, which showed self-compatibility, because self-pollen tubes were not arrested in the style, moreover abnormal post-zygotic development in ovary caused seed abortion in the cultivar. Herein we constructed a cDNA library from flowers of 'Zigui shatian' pummelo and identified one RNase gene fragment. The full length of cDNA sequence of this gene, with an open reading frame of 834 bp, was isolated by 5'-RACE method. The gene, named as CgSL2, contained five conserved regions and two histidine residues essential for RNase activity. Phylogenetic analysis indicated that CgSL2 was mostly similar to AhSL28, an S-like RNase from Antirrhinum. Southern hybridization verified CgSL2 existed in the genome as multiple copies. qRT-PCR and RT-PCR analysis showed that the expression of CgSL2 was not tissue-specific. The expression of CgSL2 was down-regulated during senescence of stem, petal, style and stamen, whereas up-regulated during ovary senescence. Further in situ hybridization of CgSL2 in the ovary during the balloon stage to anthesis stage also showed that it dramatically increased in mature flower, consistent with qRT-PCR and RT-PCR results. These findings suggested that CgSL2 might play an important role during ovary senescence.
“秭归白沙甜柚”是“沙糖柚”的自然变异株,表现为自交亲和,自花花粉管在花柱中未被阻止,并且胚后发育异常导致种子败育。本研究以‘秭归白沙甜柚’为试材,构建了柚花花蕾 cDNA 文库,克隆了一个 RNase 基因片段。通过 5’-RACE 法分离得到该基因全长 cDNA,其开放阅读框为 834bp。该基因被命名为 CgSL2,包含 5 个保守结构域和两个对 RNase 活性至关重要的组氨酸残基。系统进化分析表明,CgSL2 与金鱼草 AhSL28 具有较高的同源性,AhSL28 是一种 S 型 RNase。Southern 杂交验证 CgSL2 以多个拷贝的形式存在于基因组中。qRT-PCR 和 RT-PCR 分析表明 CgSL2 的表达无组织特异性。CgSL2 在茎、花瓣、花柱和雄蕊衰老过程中表达下调,而在子房衰老过程中表达上调。进一步对花囊中 CgSL2 的原位杂交分析表明,它在成熟花中显著增加,与 qRT-PCR 和 RT-PCR 结果一致。这些结果表明 CgSL2 可能在子房衰老过程中发挥重要作用。