芒果畸形病病原菌Fusarium mangiferae的快速分子检测

芒果畸形病是芒果上的重要病害之一,由镰孢菌侵染引起,其中以Fusarium mangiferae为主要致病菌.该病害诊断困难,且难于有效控制,因此,一旦发生则对芒果生产造成严重威胁.研究基于ISSR分子标记技术,从50条已知引物中筛选得到一条目的引物UBC 888,该引物可稳定扩增出大小为479bp的F. mangiferae特异性条带(GenBank Accession No. KJ526382).根据获得的特异性片段序列设计引物,成功地将ISSR标记转化为SCAR标记,并获得一对SCAR特异性引物(W342,W1772)和一段大小为1 376bp的特异性扩增片段(GenBank Accession No. KJ526383).通过优化特异性引物扩增条件,获得最适退火温度,构建芒果畸形病病原菌F. mangiferae的快速分子检测技术.此技术操作简单,特异性强,可检测真菌DNA的含量最低为10pg,适用于F. mangiferae和田间带菌芒果组织高灵敏度快速检测,为芒果畸形病的早期诊断和及时预防提供可靠理论依据和技术方法.

Rapid molecular detection of Fusarium mangiferae,the pathogen of mango malformation disease

Mango malformation disease (MMD), caused by Fusarium, is one of the most destructive diseases on mango, imposing continuous threat on future mango industries. The disease is difficult to diagnose and short of effective control methods. Based on ISSR molecular marker, a purpose primer (UBC 888) was selected from 50 primers, and a stable and specific band of 479bp (GenBank Accession KJ526382) was generated. According to the specific sequence, the authors designed a pair of SCAR primers (W342, W1772) and got a specific gene fragment of 1 376bp (GenBank Accession KJ526383), and the ISSR molecular markers were successfully transformed to SCAR ones. An effective detection technique of mango malformation disease was established based on this pair of primers. The established technique is simple, of high specificity and sensitivity to the pathogen both in vitro and in vivo, with the lowest detecting limit of 10pg fungal DNA. This assay provided a theoretical basis and technical method for early diagnosis and proper prevention of mango malformation disease.