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为了挖掘花生品质性状相关功能基因,本研究以花生品种花育17号为试材,根据c DNA文库中已知的FUSCA3基因全长序列设计引物,通过RT-PCR克隆到该基因,命名为Ah FUSCA3。Ah FUSCA3全长为1 532 bp,开放阅读框ORF为1 134 bp,对应的基因组序列3 892 bp,由6个外显子和5个内含子组成,内含子剪接方式符合GT-AG剪接规则。根据编码区预测Ah FUSCA3编码一条378个氨基酸组成的多肽,预测分子量为42.3 k D,等电点为5.58。预测该基因编码的蛋白含有B3保守结构域,可能定位于细胞核中。蛋白序列比对和进化树分析表明,花生与大豆(Glycine max)、菜豆(Phaseolus vulgaris)、鹰嘴豆(Cicer arietinum)和苜蓿(Medicago truncatula)等豆科植物中的FUSCA3序列相似性最高,亲缘关系最近。荧光定量PCR结果显示,Ah FUSCA3基因表达量在花生荚果、种子发育的过程中呈现先增后降的趋势,推测Ah FUSCA3基因在花生胚胎形成和发育过程中发挥重要作用。本研究为阐明花生胚胎发育过程和油脂代谢机制提供了理论基础,为花生品质改良育种提供了新的基因资源。
In order to excavate the functional genes related to the quality traits of peanut, we used the peanut variety Huayu 17 as the test material and designed the primer based on the full-length sequence of the FUSCA3 gene known in the c DNA library. The gene was cloned by RT-PCR and named as Ah FUSCA3. The full length of Ah FUSCA3 was 1 532 bp, the open reading frame ORF was 1 134 bp, and the corresponding genomic sequence was 3 892 bp, which consisted of 6 exons and 5 introns. Intron splicing was in line with GT-AG splicing rule. According to the coding region predicted Ah FUSCA3 encoding a polypeptide of 378 amino acids, the predicted molecular weight of 42.3 kD, isoelectric point of 5.58. The protein encoded by this gene is predicted to contain a B3 conserved domain that may be localized in the nucleus. Protein sequence alignment and phylogenetic tree analysis showed that peanut had the highest similarity to FUSCA3 in legumes such as Glycine max, Phaseolus vulgaris, Cicer arietinum and Medicago truncatula, Recent relationship. Fluorescence quantitative PCR results showed that Ah FUSCA3 gene expression increased first and then decreased in peanut pod and seed development, suggesting that Ah FUSCA3 gene plays an important role in peanut embryogenesis and development. This study provides a theoretical basis for elucidating the embryonic development and the mechanism of lipid metabolism in peanut and provides a new genetic resource for improved quality breeding of peanut.