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薄壁细胞次生壁木质化是石细胞形成的关键,石细胞团聚合主要受木质素单体组分及官能团含量的影响。阿魏酸5-羟化酶(F5H)是木质素合成途径中的一种关键酶,主要调控紫丁香基木质素(S木质素)单体的生物合成。本研究以砀山酥梨果实为材料,利用RT-PCR和RACE技术,得到砀山酥梨F5H全长c DNA,命名为Pb F5H(Gen Bank登录号为KC852907)。Pb F5H全长1 978 bp,其开放阅读框为1 551 bp,编码516个氨基酸,推测蛋白分子量是58.4 k Da,等电点为6.49。进化树分析表明,砀山酥梨F5H蛋白与其他物种具有很高同源性,与苹果的相似性达到96%。相关性分析表明,砀山酥梨木质素含量与石细胞含量和Pb F5H表达量高度正相关。且砀山酥梨Pb F5H参与调控梨果实S木质素单体的合成,影响木质素G/S比值。
The lignification of parenchyma cell secondary wall is the key to the formation of stone cells. The agglomeration of stone cells is mainly affected by the content of lignin monomer and functional groups. Ferulic acid 5-hydroxylase (F5H) is a key enzyme in the lignin biosynthesis pathway and mainly regulates the biosynthesis of lilac-based lignin (S-lignin) monomers. In this study, the fruit of Dangshan Suli pear was used as material. The full-length cDNA of F5H from Dangshan pear was obtained by RT-PCR and RACE, and named as Pb F5H (Gen Bank Accession No. KC852907). The full length of Pb F5H was 1 978 bp with an open reading frame of 1 551 bp encoding 516 amino acids. The deduced protein had a molecular weight of 58.4 kDa and an isoelectric point of 6.49. Phylogenetic tree analysis showed that the F5H protein of Dangshan Pear had high homology with other species, and its similarity with apple reached 96%. Correlation analysis showed that the lignin content of Dangshan Pear was highly positively correlated with the content of fossilcell and the expression of Pb F5H. And Dangshan Pear Pb F5H involved in the regulation of pear fruit S lignin monomer synthesis, affecting the lignin G / S ratio.