Isolation And Some Characteristics Identification Of Pathogen Caused Mushroom Tumor On Flammulina Velutipes

Mushroom tumor on Flammulina velutipes is a new disease on edible fungus by bacterial which divided ? and ?.This disease was caused seriously yield losses which rose to more than 80%during serious time.Once infected by the pathogen,F.velutipes mycelia form nodular lesions which stops the formation of primordia and fruiting bodies.Around 2010,this disease was first identified in the off-season cultivation areas of Sichuan,such as Hongyuan,Jiajinshan and Songpang.Since then,factory cultivated white F velutipes was also found to be affected in Sichuan,Fujian and Henan,Previously,there were no reports about the disease and pathogenie bacteria,as well as the prevention and control measures.Therefore,The present study was aimed at identifying the pathogen confirming its pathogenisity following Koch's Postulates,characterizing it using morphological,physiological,biochemical and molecular features,and screening of resistant strains and antimicrobial agents,and exploreing the pathogenic bacteria influence on other mushroom.The main research results of this paper included the following five aspects:1.Symptoms of mushroom tumor and islation pathogenicThis research take the way of field observation and laboratory vaccination records the happening of the disease and the development situation,that found two kinds of symptoms similar but different diseases,respectively named "mushroom tumor ? "and "mushroom tumor ?".For ?,tumor arrangement density,particle volume smaller,darker,produce brown liquid,and mushroom tumor ? is yellow-white,milky white liquid.Using NA medium and PDA medium Separately crossed separation of bacteria and fungi of seven disease sample substrate surface bacteria tumor,cultivation medium of junction between disease and health,internal cultivation substrate three parts,which got 68 bacteria strains and 29 fungi strains.The amount of bacteria in different parts as follows:mushroom tumor>junction between disease and health>stroma.The number of fungi and distribution were no obvious difference,and 21 of 29 fungi strains were F.velutipes.8 fungi strains of mixed bacteria only distributed in mushroom tumor and junction between disease and health,and the mixed bacteria number of mushroom tumor was more than unction between disease and health.2.Detection of mushroom tumor pathogens pathogenicSichuan gold 3 strains as the test strains,the 68 strains of bacteria and 8 strains of fungi what were no F.velutipes fungi were proved by Koeh's postulates.10 strains of bacterial pathogen which caused F.velutipes morbidity were determined by back inoculationt two times,their numbers were HY1-4?HY2-1?HY2-7?HY2-9?HY3-5?HY3-7?HY4-4?HY5-3?HY6-1?HY7-2.HY2-1 et al 5 bacteria strains were produced representative mushroom tumor ? after back inoculation,HY5-3 et al 5 bacteria strains were produced representative mushroom tumor ? after back inoculation,and all other separated stains of bacteria and fungi cannot eause F.velutipes morbidity.3.Identification of mushroom tumor pathogenic bacteriaIdentification by colony morphology and molecular biology,HY2-1?HY2-7?HY3-7?HY6-1?HY7-25 were the same species bacteria,HY1-4?HY2-9?HY3-5?HY4-4?HY5-3 were another same species bacteria.Meanwhile,we analysised physiological and biochemical property of the representative strains HY2-1 and HY5-3in these two species pathogens bacteria.The results showed that two strains were beige colonies,the gram negative bacilli,their catalase and oxidase were positive,but fermentation,gelatin liquefaction,indole test were negative.HY2-1 shouldn,t use of citrate,and HY5-3 was positive.the 16S RNA sequence of HY2-1 and HY5-3 were 100%with Ochrobactrum thiophenivorans and Ochrobactrum pseudogrignonense.4.Some characteristics research for the two pathogensThe growth pH range of them was 5-9,and they can use of yeast powder and peptone and acetic acid etc.23 kinds of carbon source.By comparing homologous,Two pathogenic bacteria strains were back inoculated to 14 speciess F.velutipes such as Sichuan 3 and Yinzheng for observation of incidence status and its degree.Two strains of pathogenic were respectively proeessed analysis of drug resistance by minocycline and other 30 kinds of drug susceptibility paper.The results showed that the F21 of had good inhibitory effect to two pathogenic bacteria strains.Yinzheng was resistance to pathogen HY5-3 performance,but no resistance to pathogenie HY2-1.The remaining 12 strains had no resistance to two kinds of pathogenic bacteria.Minocycline,ofloxacin,ciprofloxacin,cotrimoxazole four antibiotics had good inhibitory effect to two pathogenic bacterias.5.Influence of the pathogenic bacteria on other edible and medicinal fungiTwo strains of pathogenic bacteria strains were back inoculated to 20 kinds of edible and medicinal fungi like Pleurotus ostreatus,Lentinus edodes for observation of incidence status and its degree.The results show that both strains of pathogenic bacteria could not Iead to other edible and medicinal fungi nodulation and produced normally,two strains of pathogenic bacteria showed high host specificity.This study first described the F.velutipe mushroom tumor ? and ?,and the pathogenic bacteria to were respectively determined to O.Thiophenivorans and O.Pseudogrignonense.The pathogenic bacteria of physiological and biochemical characteristics and screening of antimicrobial agents provided theory basis for this kind of disease prevention control,the screening of resistant strains were prepared for late breeding work.