ABSTRACT
Pseudomonas aeruginosa MCCB 123 was grown in a synthetic medium for β-1,3 glucanase production. From the culture filtrate, β-1,3 glucanase was purified with a molecular mass of 45 kDa. The enzyme was a metallozyme as its β-1,3 glucanase activity got inhibited by the metal chelator EDTA. Optimum pH and temperature for β-1,3 glucanase activity on laminarin was found to be 7 and 50 °C respectively. The MCCB 123 β-1,3 glucanase was found to have good lytic action on a wide range of fungal isolates, and hence its application in fungal DNA extraction was evaluated. β-1,3 glucanase purified from the culture supernatant of P. aeruginosa MCCB 123 could be used for the extraction of fungal DNA without the addition of any other reagents generally used. Optimum pH and temperature of enzyme for fungal DNA extraction was found to be 7 and 65 °C respectively. This is the first report on β-1,3 glucanase employed in fungal DNA extraction.
Subject(s)
DNA, Fungal/chemistry , DNA, Fungal/isolation & purification , Glycoside Hydrolases/chemistry , Molecular Weight , Polysaccharides/chemistry , Pseudomonas aeruginosa/enzymology , Substrate Specificity , TemperatureABSTRACT
Objective To control stem blight disease of Schizonepeta tenuifolia caused by Phytophthora nicotianae.Methods The antagonist effect of 13 Trichoderma strains(including T.viride and T.hamianum)was evaluated upon mycelia growth of P.nicotianae.Trichoderma strains with high antagonistic activities against the pathogen were used to control stem blight of S.tenuifolia in the field.Results Of 13 Trichoderma strains tested,T.viride strain M3 showed maximum mycelia growth inhibition(83.2%)to the pathogen,followed by T.viride strain Tv04-2(78.2%)and then T.harziamum strain ThB(65.0%),in vitro.Fungal cell wall degrading enzymes,protease,and β-1,3-glueanase were analyzed qualitatively and quantitatively in further study.T.viride strains M3,Tv04-2,and T.harzianum strain ThB efficiently against P.nicotianae were used to control stem blight of S.tenuifolia in the field,and T.viride strain M3 showed the best biocontrol potential.Conclusion Trichoderma spp.can be used as alternatives of pesticides to control stem blight,one of the serious soilhome diseases of S.tenuifolia caused by P.nicotianae.However,though T.viride strains Tv04-2 aad T.harzianum strain ThB are also highly against P.nicotianae in vitro,the controlling efficacy of them on stem blight disease is not as excellent as T.viride strains M3 in the field.