Visual Detection Of Diaporthe Phaseolorum(Cke.&Ell.)Sacc.Var. Meridionalis Morgan-Jones(DPM)and Diaporthe Phaseolorum Var. Caulivora(DPC)Using Loop-mediated Isothermal Amplification (LAMP)

Soybean stem canker is one of the most significant soybean diseases and limits the soybean production worldwide. There are southern stem canker, which is caused by Diaporthe phaseolorum var. meridionalis (DPM), and northern stem canker, which is caused by Diaporthe phaseolorum var. caulivora (DPC). DPM and DPC belong to Ascomycotina, Sphaeriales, Diaper thaceae, Diaporthe, which cause the Diaporthe/Phomopsis complex of soybean whith Diaporthe phaseolorum var. sojae and Phomopsislongi collaHobbs. Soybean southern stem canker and soybean northern stem canker have not been reported in China. With the increasing amount of soybean traded with different countries, there are some risk that the DPM and DPC which are quarantine pathogens in China would import into china through the soybean trade between China and other countries. Suitable detection methods using a rapid, cost-effective and efficient assay are required for the control of these pathogens.At present, detection technologies of DPM and DPC are not very mature in port, even if can detect, but also takes a long time and manpower, material and equipment required for the experimental conditions, also have very high demands. Suitable detection methods using a rapid, cost-effective and efficient assay are required for the port quarantine of these pathogens. In our study, we use an alternative amplification method that developed by Zheng Xiaobo and Shen Hao to detect DPM and DPC in the absence of a thermal cycler, to establish a rapid detection technology of DPM and DPC in port.To indicate that even the sample is consist of soil microorganisms, bean, broken leaves soil and other components, our technology can successfully detect the existence of DPM and DPC, we tested the sensitivity by using Lamp assay to detect every amount gradient of spores which added in soybean residue (without DPM or DPC). Then the result implicated that this LAMP assay can detect at least 10 spores of DPM and 50 spores of DPC in lOg soybean residue.Then we use LAMP assay to detect 11 soybean residue samples that offered by Jiangsu Entry and Exit Inspection and Quarantine Bureau. DPC has been detected in 3 samples, and then DPM has not been detected.These samples that have DPC are isolated and purified. And finally we isolated DPC isolation from 2nd sample (Brazil,201304005).The actual test results show that:LAMP is an applicable, simple and quick detection method with high detection sensitivity to detect soybean northern and southern stem canker on the port. In our study, we successfully established a LAMP-based technology system for rapid detection of DPM and DPC for inspection and quarantine of imported soybeans.