南洋楹溃疡病菌巢式PCR快速检测

由可可毛色二孢菌（Lasiodiplodia theobromae）引致的溃疡病是目前威胁南洋楹健康生产和品质的重要病害。快速、准确检测病原菌是进行病害有效防控的基础。本研究用南洋楹溃疡病菌翻译延伸因子（EF 1-α）编码基因上保守靶基因区域的序列设计特异性引物EF-AF/AR。利用EF 1-α编码基因通用引物EF-688F/986R和特异性引物EF-AF/AR组合进行巢式PCR扩增,获得264 bp的单一条带,灵敏度检测最低限度为1 fg/μL。利用建立的巢式PCR方法对林间疑似溃疡病的病样进行检测,能够特异性地检测到L. theobromae。本研究建立的巢式PCR检测方法准确、特异且灵敏性高,可为南洋楹溃疡病的早期诊断和及时防控提供基础的理论和实践依据。

Establishment of a Nested-PCR Detection System of Lasiodiplodia theobromae Causing Falcataria moluccana Stem Canker Disease

A rapid nested-PCR detection system for Falcataria moluccana stem canker disease was established. The outer pair of primers EF-688F/986R and the inner pair of primers EF-AF/AR were designed based on the EF 1-α gene sequences of Lasiodiplodia theobromae. The established specific nested-PCR could amplified a single product of 264 bp with annealing temperature of 63 ℃, reaction cycles of 37. The lowest detectable concentration was 1 fg/μL. L. theobromae could be specifically detected by nested-PCR from the diseased plant samples. The establishment of rapid, sensitive nested-PCR detection system of L. theobromae might have significance in early diagnosis, and disease control of F. moluccana stem canker.