Effects Of Ectomycorrhiza Fungi And Pseudomonas Fluorescens To Pinus Tabulaeformis Growth And Disease Resistance

Using the polymerase chain reaction (PCR) - DGGE technical we analyse the relationship of soil fungal diversity, soil enzyme activity, the moisture content and, etc of pinus tabulaeformis rhizosphere in different regions of Huanglong mountain forestry. Using plate confrontation such indoor test and the potted trial of pinus tabulaeformis seedlings, we reveals the effect of ectomycorrhiza fungi and mycorrhizal helper bacteria - fluorescence pseudomonas to pinus tabulaeformis growth and disease resistance. Made the following main conclusions:1. Soil enzyme activities and fungal community diversity in the rhizosphere of Pinus tabulaeformis plantationThe research of the soil enzyme activities and the fungal community diversity in the Huanglong mountain forestry showed that the enzymatic activities of urease, alkaline phosphatase, polyphenoloxidase and hydrogenperoxidase were bottom higher than top, nightside higher than sunnyside, and enzymatic activity of roadside were always between others for its special environment. Sketch map of DGGE showed that the fungal community in the rhizosphere of Pinus tabulaeformis was similarity, but different soil sample have different position, brightness and quantity of DGGE strip, it means the type, biomass and diversity of fungi were different. The diversity of fungi was rich and distribute characteristics of diversity were the same with the enzyme activity distribution characteristics.Correlation analysis between soil enzyme activities, fungal community diversity and soil water content showed that except hydrogenperoxidase, the rest enzyme activities and fungal community diversity is a significant positive correlation (P<0.05); Soil water content and fungal diversity is a significant positive correlation (P<0.05); And except polyphenoloxidase, the rest enzyme activity and soil water content is a significant positive correlation (P<0.05); Except hydrogenperoxidase, the rest of enzyme activity is a significant positive correlation with each other (P<0.05); There is no significant correction between soil pH and enzyme activities (P>0.05).2. The resistant effect of ectomycorrhiza fungi to Rhizoctonia solaniThe confront experiment of ectomycorrhiza fungi and pathogen show that Suillus lactifluus(SL), Boletus luridus Schaeff ex Fr. (BL), MaBo, Tiantaishan5(TTS-5) were all have antagonism effect to Rhizoctonia solani, and the highest inhibition rate were SL can be 66.89%, the lowest one TTS-5 were 49.60%. In the interact region, the pathogen hypha were aging, deformation and can not normal growth by SL and TTS-5. Combining the morphological and molecular biology methods to identify the bacteria TianTaiShan 5, the result showed that, it is one of Schizophyllum.3. Effects of Pseudomonas fluorescens to ectomycorrhiza fungi and Rhizoctonia solaniGrowing promote experiment of Pseudomonas fluorescens to ectomycorrhiza fungi showed that: P. fluorescens HDY-8, HDY-9, HDY-20, HDY-35 in different concentration have different effect to SL and TTS-5 with diameter and biomass. P. fluorescens 20, HDY- 35 were the best promote one to SL; Except HDY- 35, the rest one can all promote the grow of TTS-5. The plate confront cultivation of Pseudomonas fluorescens and pathogen found that only HDY- 20 had some inhibition to Rhizoctonia solani. HDY- 9 and HDY-35 were barely resist of pathogen, and HDY- 8 even showed promote effect to pathogen.P. fluorescens 20 was not noly have promote effect to ectomycorrhiza fungi, but also have resistant effect to Rhizoctonia solani, was the best bacterial strain for promoting ectomycorrhiza fungi, prevent and cure Rhizoctonia solani.4. Effects of ectomycorrhiza fungi and Pseudomonas fluorescens to Pinus tabulaeeformis growth and resistant of damping-offAccording to the indoor experiment, we decided to use ectomycorrhiza fungi Suillus lactifluus(SL)and Schizophyllum sp.(TTS-5). Under the potted experiment, Pinus tabulaeeformis seedlings were inoculated ectomycorrhiza fungi(SL/TTS-5), inoculated Pseudomonas fluorescet(PF), simultaneously inoculated ectomycorrhiza fungi and Pseudomonas fluorescent (SL+PF/TTS-5+PF), the measure result of seedling growth, physiological character and damping-off occurrence showed that: inoculated seedlings were higher than uninoculated control in the survival, growth and root activity of seedling, and the both inoculate have the best effect, its seedling survival, seedling height, stem base, above dry weight , gross dry weight and root activity were increased by 22.10%, 36.16%, 44.86%, 124.14%, 161.54% and 100%. Both inoculated treatments can improve the soil enzyme activity and soil nutrients, except soil polyphenoloxidase, Correlation analysis shown that there were significant relationship between soil enzyme activity and soil nutrients. After inoculated ectomycorrhiza fungi, inoculated Pseudomonas fluorescet, simultaneously inoculated ectomycorrhiza fungi and Pseudomonas fluorescent or uninoculated , The MDA content of Pinus tabulaeeformis seedlings was ordered: CK>PF>SL>SL+PF,CK>PF>TTS-5>TTS-5+PF, SOD, CAT and PAL activity was SL+PF>TTS-5+PF>SL>TTS-5>PF>CK, SL+PF has the best increase level except SOD, its CAT and PAL activities were increased by 76.18% and 112.64% respectively. Incidence and disease index of inoculate plants were lower than uninoculate ones, SL+PF still has the best effect than others, its incidence and disease index were decreased by 60.94% and 70.55%. we can see that inoculated mycorrhizal fungi can effective improve soil enzyme activity, soil nutrients and enhance plant antioxidase activities, thereby promote the growth of seedlings and resist the damping-off happen , Pseudomonas fluorescens can help mycorrhizal fungi enhance this function.