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CRN(crinkling and necrosis-inducing protein)蛋白是一类卵菌特有的效应分子,然而目前对其功能和分子机制知之甚少。为分析CRN效应分子在辣椒疫霉(Phytophthora capsici)与植物互作中的作用,本研究利用前期转录组测序结果,对辣椒疫霉基因组数据库进行序列比对分析,获得了7个CRN编码基因的全长序列;采用RT-PCR方法分析其在辣椒疫霉生长发育阶段(菌丝、游动孢子囊、游动孢子、萌发的休止孢)和侵染寄主阶段(本氏烟(Nicotiana benthamiana)灌根1.5、3、6、12、24、36和72 h后)的转录水平,发现3个基因在辣椒疫霉生长发育阶段和侵染阶段上调表达;将这3个基因克隆测序,发现其编码蛋白均含有保守的LFLAK和HVLVVVP基序;在本氏烟上的基因瞬时表达结果表明,Pc559084和Pc570403能够抑制由致病疫霉elicitin蛋白INF1或老鼠促凋亡蛋白BAX诱发的植物程序性细胞死亡,并且Pc559084能够促进辣椒疫霉侵染植物。这些研究结果为理解CRN效应分子在辣椒疫霉致病过程中的作用提供了重要的实验数据。
CRN (crinkling and necrosis-inducing protein) is a type of oocyte-specific effector, however, little is known about its function and molecular mechanism. In order to analyze the interaction between CRN effector molecules and Phytophthora capsici in plants, the sequence analysis of genomes of Phytophthora capsici was carried out by using the results of pre-transcriptome sequencing. Seven CRN-encoding genes (Nicotiana benthamiana) in the growth and development stages of Phytophthora capsici (mycelium, zoospore, zoospores, spore germination) and the infection host stage (Nicotiana benthamiana) by RT-PCR. 3, 6, 12, 24, 36 and 72 h after root development), three genes were found to be up-regulated during the growth and development of Phytophthora capsici and the stage of infection. The three genes were cloned and sequenced and found to encode The proteins contained conserved LFLAK and HVLVVVP motifs. Transient expression of genes in N. benthamiana showed that Pc559084 and Pc570403 could inhibit plant programmed cell death induced by Phytophthora infestans elicitin protein INF1 or mouse pro-apoptotic protein BAX , And Pc559084 can promote Phytophthora capsici infestation of plants. These findings provide important experimental data for understanding the role of CRN effector molecules in the pathogenesis of Phytophthora capsici.