Department of Agricultural Biotechnology and Center for Fungal Pathogenesis, Seoul National University, Seoul 151-921, Republic of Korea.
Fungal Biol. 2011 Dec;115(12):1290-302. doi: 10.1016/j.funbio.2011.09.006. Epub 2011 Oct 7.
The homothallic ascomycete fungus Gibberella zeae is an important pathogen on major cereal crops. The objective of this study was to determine whether meiotic silencing occurs in G. zeae. Cytological studies demonstrated that GFP and RFP-fusion proteins were not detected during meiosis, both in heterozygous outcrosses and homozygous selfings. The deletion of rsp-1, a homologue used for studies on meiotic silencing of Neurospora crassa, triggered abnormal ascospores from selfing, but outcrosses between the mutant and wild-type strain resulted in some ascospores with mutant phenotype (low occurrence of ascus dominance). When the ectopic mutants that carried an additional copy of rsp-1 were selfed, they primarily produced ascospores with normal shape but a few ascospores (0.23 %) were abnormal, in which both endogenous and ectopically integrated genes contained numerous point mutations. The ectopic mutants showed low occurrence of ascus dominance in outcrosses with strains that carried the wild-type allele. Approximately 10 % of ascospores were abnormal but all of the single-ascospore isolates produced normal-shaped ascospores from selfing. However, no ascus dominance was observed when the mutants were outcrossed with a sad-1 deletion mutant, which lacks the putative RNA-dependent RNA polymerase essential for meiotic silencing in N. crassa. All results were consistent with those generated from an additional gene, roa, required for ascospore morphogenesis. This study demonstrated that G. zeae possesses a functional meiotic silencing mechanism which is triggered by unpaired DNA, as in N. crassa.
同宗配合的子囊菌禾谷镰孢菌是主要谷类作物的重要病原体。本研究的目的是确定减数分裂沉默是否发生在 G. zeae 中。细胞学研究表明,在异交和自交的杂合子中,GFP 和 RFP 融合蛋白在减数分裂过程中均未检测到。rsp-1 的缺失,一个用于研究粗糙脉孢菌减数分裂沉默的同源物,导致自交产生异常的子囊孢子,但突变体和野生型菌株之间的异交导致一些子囊孢子具有突变表型(低出现子囊优势)。当携带额外的 rsp-1 拷贝的异位突变体自交时,它们主要产生正常形状的子囊孢子,但少数子囊孢子(0.23%)异常,其中内源和异位整合的基因都含有大量点突变。异位突变体在与携带野生型等位基因的菌株进行异交时,表现出低的子囊优势发生率。大约 10%的子囊孢子异常,但所有单孢子分离株自交产生正常形状的子囊孢子。然而,当突变体与缺乏粗糙脉孢菌减数分裂沉默所必需的假定 RNA 依赖性 RNA 聚合酶的 sad-1 缺失突变体进行异交时,没有观察到子囊优势。所有结果与另一个基因 roa 的结果一致,该基因对于子囊孢子形态发生是必需的。本研究表明,G. zeae 具有一种功能齐全的减数分裂沉默机制,该机制由未配对的 DNA 触发,与粗糙脉孢菌相似。