The Inhibition Activity Of Small Molecular Substances Of Pseudomonas Monteilii On TMV And PVY

In order to develop antivirus products, strain4A1with high efficiency and high activity againstTMV and PVY was selected from healthy tobacco(Nicotiana tabacum var. Zhongyan100) field fromthe area infected TMV in Zhucheng. The antiviral rate of4A1is up to98%by half-leaf method in lab.Real-time quantitative PCR was used to test the inhibition of TMV and PVY for further verifying theanti-viral activity. The strain was identified by the physiological and biochemical and molecularidentification. Sequence analysis results showed that the nucleotide sequence of the16S rDNA of thestrain had99%identity with that of Pseudomonas monteiliis. The strain4A1is gram-negative. Enzyme,starch hydrolysis reaction such as VP and indole were negative, Gelatin, liquefaction, glucose responsewere positive and can move.4A1was identified as P. monteilii based on the above physiological,biochemical and molecular test results. Field trials were handled. Treatment I mixed the virus juice withfermentation for2h can effectively inhibit most viral infection. TMV incidence rate was5.3%. PVYincidence rate was7.7%. Disease index respectively was0.8and2.9. Those are much lower than othertreatments.The results of strains’ culture medium and fermentation conditions showed conditions were asfollowing: the culture temperature:28℃, the culture time48h, inoculation amount3%, initial pH=10,ventilation volume100mL/250mL, glucose and corn powder as the carbon source, yeast extract andpeptone as mixed nitrogen sources. It may reach the ideal effect.Active constituents from bacterial strain4A1was isolated and purified through pretreatment,solvent extraction, D-101macroporous adsorption resin and silica gel chromatography adsorption.Liquid-liquid extraction was proved that the liquid phase has anti-TMV activity through biologicaldetermination. We analyze the chemical composition and structure identification by GC/MS. Weseparated4kinds of compounds which identified by GC/MS. There were2,4,6(1H,3H,5H)–Pyrimidinetrione,5-butyl-5-ethyl-1,3-bis(trimethylsilyl); Di-n-octyl phenyl phosphate;1,2-Benzenedicarboxylic acid, mono(2-ethylhexyl) ester; Hexanedioic acid, bis(2-ethylhexyl) ester.We made study about the mechanism of small molecular against TMV and PVY. The resultsshowed that TMV particles can be inactivated by the activity substances from4A1broth.Polyacrylamide gel electrophoresis demonstrated active material can crack virus coat protein. the virusparticle lose infectivity.