Medeiros L V, Maciel D B, Medeiros V V, Houllou Kido L M, Oliveira N T
Laboratório de Genética Molecular de Fungos, Universidade Federal de Pernambuco, Recife, PE, Brazil.
Genet Mol Res. 2010 Apr 13;9(2):661-73. doi: 10.4238/vol9-2gmr725.
Colletotrichum gloeosporioides is an important pathogen for a great number of economically important crops. During the necrotrophic phase of infection by Colletotrichum spp, the degradative enzymes of plant cell walls, such as pectate lyase, clearly increase. A gene pelB that expresses a pectate lyase was identified in isolates of C. gloeosporioides in avocado pathogens. Various molecular studies have identified a kind of specialization of C. gloeosporioides isolates with specific hosts; however, there have been no studies of this gene in isolates from hosts other than avocado. The same is true for other species of Colletotrichum. We examined genetic variability in order to design primers that would amplify pelB gene fragments and compared the products of this amplification in C. gloeosporioides isolates from different hosts. Genetic variability was assessed using ISSR primers; the resultant data were grouped based on the UPGMA clustering method. Primers for the pelB gene were designed from selected GenBank sequences using the Primer 3 program at an annealing temperature of 60 degrees C and product amplification of nearly 600 bp. The ISSR primers were efficient in demonstrating the genetic variability of the Colletotrichum isolates and in distinguishing C. gloeosporioides, C. acutatum and C. sublineolum species. The gene pelB was found in C. gloeosporioides, C. acutatum and C. sublineolum. Amplified restriction fragments using MspI did not reveal differences in pelB gene structure in isolates from the three different host species that we investigated.
胶孢炭疽菌是许多经济作物的重要病原菌。在炭疽菌属感染的坏死营养阶段,植物细胞壁的降解酶,如果胶裂解酶,会明显增加。在鳄梨病原菌的胶孢炭疽菌分离物中鉴定出一个表达果胶裂解酶的基因pelB。各种分子研究已经确定了胶孢炭疽菌分离物对特定宿主的一种专一性;然而,除了鳄梨之外,尚未对来自其他宿主的分离物中的该基因进行研究。炭疽菌属的其他物种也是如此。我们研究了遗传变异性,以便设计能扩增pelB基因片段的引物,并比较来自不同宿主的胶孢炭疽菌分离物中该扩增产物。使用ISSR引物评估遗传变异性;所得数据根据UPGMA聚类方法进行分组。根据选定的GenBank序列,使用Primer 3程序在60℃退火温度下设计pelB基因的引物,产物扩增片段大小约为600 bp。ISSR引物能有效地显示炭疽菌分离物的遗传变异性,并区分胶孢炭疽菌、尖孢炭疽菌和亚线炭疽菌。在胶孢炭疽菌、尖孢炭疽菌和亚线炭疽菌中均发现了pelB基因。使用MspI酶切扩增片段后,未发现我们所研究的三种不同宿主物种的分离物在pelB基因结构上存在差异。