甘蓝型油菜LPAT4基因的克隆与表达

Cloning and Expression of LPAT4 Genes from Brassica napus

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,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BnLPAT4-2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的表达对PEG-4000更敏感。本试验为进一步研究油菜BnLPAT4基因功能奠定了基础。

Lysophospholipid acid actyltransferase （LPAT） is a pivotal enzyme of triacylglycerol biosynthesis and plays a key role in lipid synthesis, development of plant seeds and bio-membrane fluidity. Two LPAT4 full-length coding sequences （1143 bp and 1140 bp） were obtained by homology-based cloning, designated BnLPAT4-1 and BnLPAT4-2, respectively. Bioinformatics analysis revealed that they share the LPLAT_LCLAT1 like domain and belong to the LPLAT super family. Temporal and spatial expression results showed that BnLPAT4-1 and BnLPAT4-2 are genes of constitutive expression, with the highest expression in leaf and em-bryo, respectively. BnLPAT4-1 and BnLPAT4-2 showed different expression patterns under NaCl, PEG-4000, waterlogging, 6BA and ABA stresses. Pole difference analysis indicated that BnLPAT4-1 was regulated greatly by ABA, while BnLPAT4-2 was more sensitive to PEG-4000. These results are informative for function study of BnLPAT4 genes in Brassica napus.