Application Of Loop-mediated Isotheral Amplication (LAMP) In Rapid Detection Of Fungal Soybean Root And Stem Disease

Soybean is found in China earliest an annual herb that be cultivated over the world.There are many pathogens caused diseases of soybean.The soybean plant root and stem disease are a dangerous,recalcitrant and notorious disease and has induced losses in yield and seed quality in mainsoybean production regions over the world.It is difficult to judge which pathogens caused disease only by according symptom of incipient stage.Traditionally have been used to detect pathogens in plant material but may take a considerable amount of time,and it is easy to turn up misdiagnosis.The PCR-based methods can be completed must be in the laboratory and take a considerable amount of time.However,the methods might not be adverse to rapid detection.Therefore,there is a growing for simple and economical tests.This is the first report of LAMP detection of fungalplant disease,It possess characteristic of simpleness,rapidness,safeness and lowness-cost.The detection system had the advantages of without needing special instruments,judgement with the colour change of reaction mixture,and can be used for detecting the simple in the field.Establishment of LAMP provides a necessary new alternative method for the rapid detection of plant disease in the field.1.The Fusarium spp.and Rhizoctonia solani?Macrophomina phaseolina LAMP detected by the seven LAMP system in our laboratory from 640 samples of diseased soybean plant roots were collected from Jiangsu,Anhui,Hubei and Yunnan provinces 5and purified the pathogen separations identified as Fusarium?Rhizoctonia solaniand Macrophomina phaseolina,and thendetermined their pathogenicity.The results indicate that the positive-sample ratios by the LAMP assay,respectively,20%of Fusarium oxysporuum?27%of Fusarium solani?12%of Fusarium proliferatum?7%of Fusarium equiseti?4%of Fusarium graminearum?10%of Rhizoctonia solani?13%of Macrophomina phaseolina.The cross-infection is very serious in Jiangsu,Anhui provinces,the positive-sample ratio was 31%by the LAMP assay,and the positive-sample ratioswere 6%by the two pathogens caused disease,24%by the three pathogens caused disease,Fusarium oxysporum and Fusarium solani are the dominant pathogens,Fusarium proliferatum and Fusarium equiseti are the second pathogens.The LAMP procedure is very simple and rapid which in the detection can be completed in only 1.5 to 2 h,compared with traditionally and PCR-based methods,the detection is simple,rapid,safe and low-cost.It could be potentially applied to detection of DNA of pathogens in poor-equipped laboratories.It is very conductive to directly prevent and treatment in the disease of agricultural production.2.The study designed a set method which is simple,practical use of the vacuum flask to replace PCR apparatus provides isothermal(or similar isothermal)condition to complete LAMP assay.It showed that the sensitivity of LAMP can achieve the request of LAMP detection,and can be used for detection the simple in the field.We had established a LAMP detection system(2-1b)of fungal plant disease,It possess characteristic of simpleness,rapidness,safeness and lowness-cost.The detection system had advantages of without needing special instruments,judgement with the colour change of reaction mixture,and can be used for detection the simple in the field.It could be potentially applied to detection of DNA of pathogens in poor-equipped laboratories.