甘蓝型油菜LPAT4基因的克隆与表达

植物溶血磷脂酸酰基转移酶(lysophospholipid acid actyltransferase, LPAT)是三酰甘油生物合成过程中的一个关键酶, 在脂质的合成、种子的发育以及生物膜的流动性等方面有重要作用。本研究采用同源克隆的方法, 获得LPAT4基因的2条全长CDS序列, 长度分别为1143 bp和1140 bp。生物信息学分析表明, 它们均具有LPLAT_LCLAT1样结构域, 同属于LPLAT超基因家族, 并分别被命名为BnLPAT4-1和BnLPAT4-2。时空表达分析表明, 它们均为组成型表达基因, 其中BnPAT4-1在叶中的表达量最高, 而BnPAT4-2在胚中的表达量最高。逆境分析表明, BnLPAT4-1和BnLPAT4-2在NaCl、PEG-4000、水渍、6BA和ABA的胁迫下呈现出不同的表达模式。极差分析显示, ABA对BnLPAT4-1的表达影响较大, 而BnLPAT4-2的表达却对PEG4000更敏感。为进一步研究油菜BnLPAT4基因功能奠定了基础。

Cloning and Expression Analysis of LPAT4 Gene from Brassica napus

Lysophospholipid acid actyltransferase (LPAT) is a pivotal enzyme of triacylglycerol biosynthesis, which plays a key role in lipid synthesis, development of plant seeds and bio-membrane fluidity. Through the technology of homology-based cloning, two copies of LPAT4 full-length CDS sequences (1143 bp and 1140 bp) were cloned in this study, designated as BnLPAT4-1and BnLPAT4-2, respectively. Bioinformatics analysis revealed that they shared the LPLAT_LCLAT1 like domain and belonged to the LPLAT superfamily. Temporal and spatial expression results showed that BnLPAT4-1 and BnLPAT4-2 were constitutive expression genes. Among them, the highest expression of BnLPAT4-1 was in leaf, while that ofBnLPAT4-2 was in embryo. Stresses analysis indicated that BnLPAT4-1and BnLPAT4-2 presented different expression patterns under the treatments of NaCl, PEG4000, waterlogging, 6BA and ABA. Pole difference analysis displayed that ABA had a great effect on the expression of BnLPAT4-1, while BnLPAT4-2 was more sensitive to PEG-4000.The results provided a base for the research of regulation and function of BnLPAT4 in Brassica napus.