National Research Council of Canada, Plant Biotechnology Institute, 110 Gymnasium Place, Saskatoon, Saskatchewan, S7N 0W9 Canada.
Mol Plant Pathol. 2001 May 1;2(3):159-69. doi: 10.1046/j.1364-3703.2001.00063.x.
Summary To examine the defence response in Brassica carinata we differentially screened a cDNA library made from CuCl(2)-treated (Cu) leaves. The sequence of 17 of the 27 cDNA clones examined that showed Cu-induction had a high similarity to defence genes from other plant species. Among other clones that showed higher expression in the Cu leaves were two cDNAs encoding polypeptides of 351 and 250 amino acids, designated BcCJS1 and BcCJAS1. BcCJS1 had similarity to S-adenosyl-l-methionine: salicylic acid carboxyl methyltransferase from Clarkia breweri. However, the enzyme activity was not found in extracts from E. coli expressing BcCJS1. BcCJAS1 did not show extensive similarity to any genes with known function in the databases but it did contain three regions of amino acid sequence that are frequently found in amidotransferases. A third Cu-induced mRNA, Bcp6PGL, showed very high (86%) similarity to a putative 6-phosphogluconolactonase (6PGL) from Arabidopsis thaliana. In addition to Cu induction, BcCJS1 expression was induced by methyl jasmonate (MeJA) and salicylic acid (SA), BcCJAS1 expression by MeJA, SA and abscisic acid and Bcp6PGL expression by MeJA. The expression of all three genes increased after Alternaria brassicae infection. BcCJS1 and BcCJAS1 were induced within 1 h after MeJA- but not until 3 h after SA-treatment. The expression of both genes was systemically induced after infection with a compatible or incompatible fungal pathogen. SA systemically induced only BcCJAS1. The effects of various inhibitors of signalling pathways on expression of the three genes were studied.
摘要 为了研究油菜(Brassica carinata)的防御反应,我们从 CuCl2 处理(Cu)的叶片中差异筛选 cDNA 文库。在 27 个显示 Cu 诱导的 cDNA 克隆中,有 17 个序列与其他植物物种的防御基因具有高度相似性。在其他在 Cu 叶片中表达更高的克隆中,有两个编码 351 和 250 个氨基酸的多肽的 cDNA,分别命名为 BcCJS1 和 BcCJAS1。BcCJS1 与来自 Clarkia breweri 的 S-腺苷-L-蛋氨酸:水杨酸羧甲基转移酶具有相似性。然而,在表达 BcCJS1 的大肠杆菌提取物中未发现该酶的活性。BcCJAS1 与数据库中具有已知功能的任何基因没有广泛的相似性,但它确实包含三个经常在氨基转移酶中发现的氨基酸序列区域。第三个 Cu 诱导的 mRNA,Bcp6PGL,与拟南芥的 6-磷酸葡萄糖酸内酯酶(6PGL)非常相似(86%)。除了 Cu 诱导外,BcCJS1 的表达还被茉莉酸甲酯(MeJA)和水杨酸(SA)诱导,BcCJAS1 的表达还被 MeJA、SA 和脱落酸诱导,Bcp6PGL 的表达还被 MeJA 诱导。在黄萎病菌(Alternaria brassicae)感染后,这三个基因的表达都增加了。在 MeJA 处理后 1 小时内诱导 BcCJS1 和 BcCJAS1,但在 SA 处理后 3 小时才诱导。在与相容或不相容真菌病原体感染后,这两个基因都被系统性诱导。SA 仅系统性诱导 BcCJAS1。研究了各种信号通路抑制剂对这三个基因表达的影响。