Pieterse C M, Verbakel H M, Spaans J H, Davidse L C, Govers F
Department of Phytopathology, Wageningen Agricultural University, The Netherlands.
Mol Plant Microbe Interact. 1993 Mar-Apr;6(2):164-72. doi: 10.1094/mpmi-6-164.
In order to isolate in planta-induced genes encoding putative pathogenicity factors of the late blight fungus Phytophthora infestans, a genomic library was differentially screened. For the differential hybridization, labeled first-strand cDNA synthesized on mRNA isolated from P. infestans-infected potato leaves and on mRNA isolated from the fungus grown in vitro were used as probes. This screening resulted in the isolation of the P. infestans calmodulin gene. The gene, designated calA, contains an open reading frame of 447 base pairs without introns and is unique in the P. infestans genome. The predicted amino acid sequence is 89.9-94.6% identical to calmodulins from higher eukaryotes, whereas the identity to calmodulins of higher fungi is significantly less (60.8-85.1%). Expression studies revealed that the P. infestans calA gene is constitutively expressed in in vitro grown mycelium. However, during pathogenesis on potato the level of P. infestans calmodulin mRNA is increased approximately fivefold.
为了分离植物体内由致病疫霉(Phytophthora infestans)诱发的、编码假定致病因子的基因,对一个基因组文库进行了差异筛选。对于差异杂交,以从感染致病疫霉的马铃薯叶片中分离的mRNA以及从体外培养的该真菌中分离的mRNA合成的标记第一链cDNA作为探针。该筛选导致了致病疫霉钙调蛋白基因的分离。该基因命名为calA,包含一个447个碱基对的开放阅读框,无内含子,在致病疫霉基因组中是独特的。预测的氨基酸序列与高等真核生物的钙调蛋白具有89.9 - 94.6%的同一性,而与高等真菌钙调蛋白的同一性则显著较低(60.8 - 85.1%)。表达研究表明致病疫霉calA基因在体外生长的菌丝体中组成性表达。然而,在对马铃薯致病过程中致病疫霉钙调蛋白mRNA的水平大约增加了五倍。