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利用苯丙氨酸解氨酶(PAL,phenylalanine ammonia-lyase)基因保守区域从小麦抗赤霉病材料苏麦3号中克隆获得4个PAL基因,分别命名为Ta PAL1、Ta PAL2、Ta PAL3、Ta PAL4。4个基因的开放阅读框(ORF,open reading frame)长度分别为2142 bp、2016 bp、2118 bp和2139 bp,分别编码714个、672个、706个和713个氨基酸。基因序列比对发现其相似性达到88.35%,所编码的氨基酸相似性为91.92%,氨基酸序列分析表明4个基因都包含HAL-PAL结构域及PAL结构域。通过接种禾谷镰刀菌,利用荧光定量PCR对PAL基因进行表达分析发现,4个PAL基因全部为上调表达,其中Ta PAL2、Ta PAL3和Ta PAL4最为明显。PAL基因的上调表达,说明PAL基因在小麦抵抗赤霉病菌侵染的机制中可能起着重要作用。
Four PAL genes were cloned from the wheat cultivar Sumai 3 using the conserved region of phenylalanine ammonia-lyase (PAL) gene and named as Ta PAL1, Ta PAL2, Ta PAL3, The open reading frame (ORF) of Ta4.4 genes were 2142 bp, 2118 bp and 2139 bp in length, encoding 714, 672, 706 and 713 amino acids respectively. The amino acid sequence identity of the two genes was 88.35%, and the similarity of amino acids was 91.92%. Amino acid sequence analysis showed that all four genes contained HAL-PAL domain and PAL domain. Through the inoculation of Fusarium graminearum, the expression of PAL gene was analyzed by fluorescence quantitative PCR. All four PAL genes were up-regulated, of which Ta PAL2, Ta PAL3 and Ta PAL4 were the most obvious. Up-regulated expression of PAL gene indicates that PAL gene may play an important role in the mechanism of wheat resistance to Fusarium graminearum infection.