Chadha S, Gopalakrishna T
Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Mumbai, India.
J Appl Microbiol. 2006 May;100(5):1147-53. doi: 10.1111/j.1365-2672.2006.02920.x.
To develop a diagnostic assay based on polymerase chain reaction for the detection of Magnaporthe grisea from infested rice seeds.
Primers were designed based on the nucleotide sequence of the mif 23, an infection-specific gene of M. grisea. The primers amplified target DNA from genetically and geographically diverse isolates of the pathogen. The lowest concentration of template DNA that led to amplification was 20 rhog. No PCR product was detected when DNA from other fungi was used, indicating the specificity of the primers. With this PCR based seed assay, M. grisea was detected in rice seedlots with infestation rates as low as 0.2%.
The PCR detection of M. grisea is simple, rapid, specific, sensitive and suitable for the routine detection of the pathogen in infested seeds.
Introduction of the blast fungus into new areas where it has not been previously recorded could be avoided by the detection of infested seedlots. A PCR-based seed assay could facilitate risk assessment of naturally infested rice seeds; help design management programs and optimize fungicide use.
开发一种基于聚合酶链反应的诊断检测方法，用于检测受侵染水稻种子中的稻瘟病菌。
根据稻瘟病菌感染特异性基因mif 23的核苷酸序列设计引物。这些引物能从该病原菌在遗传和地理上不同的分离株中扩增出目标DNA。导致扩增的模板DNA最低浓度为20 rhog。当使用其他真菌的DNA时未检测到PCR产物，表明引物具有特异性。通过这种基于PCR的种子检测方法，在侵染率低至0.2%的水稻种子批次中检测到了稻瘟病菌。
稻瘟病菌的PCR检测方法简单、快速、特异、灵敏，适用于受侵染种子中该病原菌的常规检测。
通过检测受侵染的种子批次，可以避免稻瘟病菌传入以前未记录过的新地区。基于PCR的种子检测方法有助于对自然侵染的水稻种子进行风险评估；有助于设计管理方案并优化杀菌剂的使用。
