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An integrative overview on the function and regulation of the PITP gene family in sugarcane

Huaying Mao  Weihua Su  Yongjuan Ren  苏亚春  Dongjiao Wang  Hanchen Tang  Liping Xu  Youxiong Que  
【摘要】:【Research background】 Sec14 p, one of the phosphatidylinositol transfer proteins(PITPs), is originally found in Saccharomyces cerevisiae that has the functions of transporting phosphatidylinositol(phosphatidylinositol, PI) and phosphatidylcholine(phosphatidylcholine, PC) and is widely present in eukaryotes. Plant Sec14-like phosphatidylinositol transfer protein has high sequence homology with yeast Sec14 p. In recent years, with the development of molecular biology and lipid genomics, more and more plant PITPs have been excavated. However, to date, the investigation on the PITP genes and their functions has not been conducted in sugarcane. 【Materials and methods】 In this study, six sugarcane Sec14-like PITP genes were mined from a sugarcane transcriptome database. The full-length c DNA sequences of ScSEC14, ScSEC14-1 and Sc SEC14 p were isolated and characterized by prokaryotic expression, spot assay, subcellular localization, transient expression, and ScSEC14 p genetic transformation in Nicotiana benthamiana. 【Result and analysis】 Phylogenetic analysis showed that six ScPITPs proteins were divided into four groups, including group I(ScSFH1 and ScSFH2), Group II(ScPATL1 and ScPATL2), Group III(ScSEC14 p) and Group V(ScSEC14 and ScSEC14-1). In the abiotic stresses of PEG, NaCl, CuCl2, CdCl2 and 4℃ treatment, most of these genes responded to PEG and NaCl stresses. ScSEC14 was located in the plasma membrane and nucleus, ScSEC14-1 and ScSEC14 p were located in the plasma membrane and cytoplasm according to subcellular localization. The spot assay showed that the recombinant proteins of ScSEC14-1 and ScSEC14 p could enhance the tolerance of Escherichia coli to drought and salt stresses. The transient expression results suggested that ScSEC14, ScSEC14-1 and ScSEC14 p genes may involve in plant immunity and exhibited an antimicrobial action to Ralstonia solanacearum and Fusarium solani var. coeruleum. The genetic transformation results showed that overexpression of ScSEC14 p increases the germination rate of transgenic N. benthamiana seeds under mannitol and NaCl stresses. The enhanced tolerance to drought and salt stresses of N. benthamiana plants overexpressing ScSEC14 p may be mainly attributed to enhanced antioxidant enzyme activity and upregulation of ROS scavenging enzymes encoding genes, osmotic stress-defense genes, and PLC and PLD genes. 【Conclusion】 These results improve our understanding of the functions of the PITP gene family in sugarcane complexes as well as identify the other members of the Sec14-like PITP gene family in sugarcane.

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