Research Institute for Biotechnology and Environment, Nong Lam University, Ho Chi Minh City, Vietnam.
Faculty of Biotechnology, Ho Chi Minh City Open University, Ho Chi Minh City, Vietnam.
Int Microbiol. 2021 May;24(2):149-156. doi: 10.1007/s10123-020-00150-0. Epub 2020 Nov 7.
PCR-based molecular approaches including RAPD (random amplified polymorphic DNA), ISSR (inter-simple sequence repeat), and SRAP (sequence-related amplified polymorphism) are commonly used to analyze genetic diversity. The aims of this study are to analyze genetic diversity of M. oryzae isolates using PCR-based molecular approaches such as RAPD, ISSR, and SRAP and to develop SCAR marker linked to the pathogenicity of rice blast fungus. Twenty Magnaporthe oryzae isolates were collected mainly from the south of Vietnam and assessed for genetic variation by RAPD, ISSR, and SRAP methods. The comparison of those methods was conducted based on the number of polymorphic bands, percentage of polymorphism, PIC values, and phylogenetic analysis. Then, sequenced characterized amplified region (SCAR) markers were developed based on specific bands linked to fungal pathogenicity of rice blast fungus, M. oryzae. The results indicated that SRAP markers yielded the greatest number of polymorphic bands (174) and occupied 51.7% with polymorphism information content (PIC) value of 0.66. Additionally, the SRAP approach showed stability and high productivity compared with RAPD and ISSR. The SCAR marker developed from the SRAP method identified the presence of the avirulence AVR-pita1 gene involving fungal pathogenicity that can break down blast resistance in rice cultivars. The consistency of SCAR marker obtained in this study showed its efficiency in rapid in-field detection of fungal pathogenicity. SCAR marker developed from SRAP technique provides a useful tool for improving the efficiency of blast disease management in rice fields.
基于聚合酶链反应（PCR）的分子方法，包括随机扩增多态性 DNA（RAPD）、简单重复间序列多态性（ISSR）和序列相关扩增多态性（SRAP），常用于分析遗传多样性。本研究旨在使用基于 PCR 的分子方法，如 RAPD、ISSR 和 SRAP 分析稻瘟病菌的遗传多样性，并开发与稻瘟病菌致病性相关的 SCAR 标记。本研究从越南南部采集了 20 个稻瘟病菌分离株，通过 RAPD、ISSR 和 SRAP 方法评估其遗传变异。基于多态带数、多态性百分比、PIC 值和系统发育分析比较了这些方法。然后，根据与稻瘟病菌致病性相关的特定带，基于测序特征扩增区（SCAR）标记开发了稻瘟病菌的 SCAR 标记。结果表明，SRAP 标记产生的多态带数量最多（174 个），多态性百分比为 51.7%，多态性信息含量（PIC）值为 0.66。此外，与 RAPD 和 ISSR 相比，SRAP 方法表现出稳定性和高生产力。从 SRAP 方法开发的 SCAR 标记鉴定了涉及真菌致病性的无毒基因 AVR-pita1 的存在，该基因可以破坏水稻品种的抗瘟性。本研究中获得的 SCAR 标记的一致性表明其在快速现场检测真菌致病性方面的效率。从 SRAP 技术开发的 SCAR 标记为提高稻田稻瘟病管理效率提供了有用的工具。
