Freeman S, Katan T, Shabi E
Department of Plant Pathology, Agricultural Research Organization, The Volcani Center, Bet Dagan, Israel.
Appl Environ Microbiol. 1996 Mar;62(3):1014-20. doi: 10.1128/aem.62.3.1014-1020.1996.
One hundred twenty isolates of Colletotrichum gloeosporioides from avocado (6 U.S. and 57 Israeli isolates) and almond (57 Israeli isolates) fruits were compared by various molecular methods and a pathogenicity assay in order to determine the genetic diversity and host specificity between and among the different populations. DNA from eight additional U.S. almond anthracnose isolates were also compared. PCR amplification of genomic DNA with four primers produced uniform banding patterns for all the Israeli almond isolates from different geographic locations in Israel. DNAs from the U.S. almond isolates were distinct from DNAs of the Israeli isolates. In contrast, the avocado isolates from Israel and the United States were more diverse, with numerous arbitrarily primed-PCR phenotypes being observed. HaeIII digestion patterns of A+T-rich DNA distinguished between the almond and avocado isolates. Southern hybridization of the repetitive nuclear-DNA element GcpR1 to PstI-digested genomic DNA of almond and avocado isolates revealed no polymorphic fragments among the almond isolates, whereas polymorphic fragments were observed among the avocado isolates. Amplification and subsequent restriction enzyme digestion of the internal transcribed spacer 4 and 5 regions between the small and large nuclear subunits of DNA encoding rRNA failed to distinguish between C. gloeosporioides isolates from a diverse host range. In artificial inoculations, avocado isolates produced various lesions on avocado and almond fruits, whereas the almond isolates infected both fruits at a lower rate.
通过多种分子方法和致病性测定,对从鳄梨果实(6株美国分离株和57株以色列分离株)和杏仁果实(57株以色列分离株)中获得的120株胶孢炭疽菌分离株进行了比较,以确定不同种群之间以及种群内部的遗传多样性和寄主特异性。还对另外8株美国杏仁炭疽病分离株的DNA进行了比较。用4种引物对基因组DNA进行PCR扩增,结果显示来自以色列不同地理位置的所有以色列杏仁分离株具有一致的条带模式。美国杏仁分离株的DNA与以色列分离株的DNA不同。相比之下,来自以色列和美国的鳄梨分离株则更加多样,观察到许多任意引物PCR表型。富含A+T的DNA的HaeIII消化模式区分了杏仁和鳄梨分离株。将重复核DNA元件GcpR1与杏仁和鳄梨分离株经PstI消化的基因组DNA进行Southern杂交,结果显示杏仁分离株之间没有多态性片段,而在鳄梨分离株中观察到了多态性片段。对编码rRNA的DNA大小亚基之间的内部转录间隔区4和5区域进行扩增并随后进行限制性酶切,未能区分来自不同寄主范围的胶孢炭疽菌分离株。在人工接种中,鳄梨分离株在鳄梨和杏仁果实上产生了各种病斑,而杏仁分离株对两种果实的感染率较低。