百合 ATP 合成酶家族基因 ATPase3 的 克隆和生物信息学分析.

【Objective】The paper aimed to effectively solve the problem of pollen pollution and cultivate male sterile Lilium using molecular biology technology. The role of ATPase3 gene, a member of the ATPase family, in the male sterility Lilium was studied, which would provide a theoretical basis for the application of this gene in the pollen-free molecular breeding of Lilium.【Method】Anthers of fertile lines and sterile lines of Liliums were used as experimental materials, and a significantly up-regulated ATPase3 gene was screened from the results of transcriptome sequencing of fertile lines. Relative expression of ATPase3 gene in different parts of Lilium was detected by fluorescence quantitative qRT-PCR during the period of microsporocyte. Full-length cDNA sequence of anther ATPase3 gene of Lilium was amplified by qRT-PCR and Race technology, and the structure and physicochemical properties of Lilium ATPase3 were analyzed by various online bioinformatics software（Prot Param, Prot Scale, NCBI Conserved Domain, SWISS-MODEL）. 【Result】The expression of ATPase3 gene in Lilium was the highest in flower organs, and the expression in anthers of fertile lines was 7 times higher than that of sterile lines. Full-length ORF of Lilium ATPase3 gene was 327 bp, encoding 109 amino acids and containing a typical conserved ATPase3 domain; Relative molecular weight of the protein was 12.14 KD and theoretical isoelectric point was 7.71. It was a hydrophilic protein with transmembrane structure, and the secondary and tertiary structures of the protein were predicted. Bioinformatics analysis showed that the predicted protein had typical characteristics of ATP synthase. Homologous sequence analysis showed that the protein had the closest homology with Arabidopsis thaliana plasma membrane ATPase3 and H[+]ATPase1, H[+]ATPase2, H[+]ATPase3, H[+]ATPase8. This protein was a plasma membrane ion transporter that provided energy in plants. It was speculated that the expression level of this protein was down-regulated in the anthers of Lilium sterile lines, which led to the male sterility of Lilium.【Conclusion】In the study, ATPase3 gene was verified to be related to male sterility in Lilium, and ATPase3 gene was successfully cloned and analyzed by bioinformatics. It provided a theoretical basis for the occurrence of male sterility in Lilium caused by the loss of plasma membrane ion transporter function which was encoded by ATPase3 gene. [ABSTRACT FROM AUTHOR]