The Study On The Diversity Of Ectomycorrhizal Fungi Of Pinus Sylvestris Var. Mongolica Litv And Mycorrhizal Biotechnology

This research including morphological and anatomical characteristic analysis of Pinussylvestris var. mongolica Litv ectomycorrhizal types;study on molecular identification tothese ectomycorrhizae.; effect of ectomycorrhizae on the growth of Pinus sylvestris varmongolica Litv seedlings and the mycorrhizal formation of tissue cultured plantlets onpinus sylvestris var. mongolica Litv Now respectively abstract writes as follow:1.Morphological and anatomical characters analysis of ectomycorrhizaeEctomycorrhizal fungus classification and identificaton shoud be put emphasis on thesymbiotic fungus organizational characteristic with tree species, because the morphologicaland anatomical feature of fungus organization is stable and it was proved to providingqualification and possiblility for ectomycorrhizal fungus classification and identifi cation.In this study, taking mycorrhizal sysbiosis as the study object, from Pinus sylvestris var.mongolica Litv starting and from the morphological and anatomical angle, studied theseectomycorrhizal types systematicly, and comprehensive characteristic of19ectomycorrhiz-al types were described using photographs and words. At the same times, on this basis,made a generalization and drawn a conclusion on ectomycorrhizal colour, morphologicalcharacters, surface structure of mantle and anatomical characters in plan views and so on;Further probed in the feasibility of ectomycorrhizal classification and identification in thelight of exterior hyphae, mycorrhizal colour, interior microstructure shuc as arrangedpatterm of mantle hyphae, mantle surface structure and anatomical structure of hyphae andrhizomorphs mainly. On the basis of this study have make a species identification on all19ectomycorrhizal types by means of molecular technology.2. Molecular identification reseach on the ectomycorrhizal root tipsThis research was on the premise of having been sorted out basal ectomycorrhizaltypes, took the mycorrhizal symbiosis root tips of Pinus sylvestris var. mongolica Litv asstudied material, extracted DNA of symbiosis, then ITS regions of DNA of identifiedectomycorrhizal fungi were amplified using the special pairs of primers ITS1F/ITS4andsequenced, the sequences determined were compared with those registered in rDNAsequence database on Internet, then identification results were obtained according tocomparing results and other biological information in DNA sequence database on Internet. At last, ectomycorrhizal fungi diversity of several trees species in Inner mongolia wereconfirmed. The results write as follow: of the total of19morphology and anatomaticaltypes of Pinus sylvestris var. mongolica Litv,19types were identified to the level of family,genus and species respectively, ectomycorrhizal types of Pinus sylvestris var. mongolicaLitv belonging respectively to family level is3, genus level is1and rest of15is identifiedto species level; At the same times, among19identified ectomycorrhizal types,15speciesbelong to Basidiomycotina and4species belong to Ascomycotina.This research have finished species identifications, as well as by using the biologicalinformation resources on Internet adequately, constructed the taxonomy report, andanalysed the compared results and taxonomy report.3. Effect of ectomycorrhizae on the growth of pinus sylvestris var. mongolica LitvseedingsIn this research,the mycorrhizae synthesis between Pinus sylvestrisvar. mongolicaLitv and Suillus granulatus O.Kurttee、Suillus grevillei Sing、Tuber liaotongense、Lactariuspubescens、Cortinarius caerulescens Fr、Russula xerampelina Cenococcum geophilum Fr、Pisolithus tinctorius Coker＆Couch.and Paxillus involulus Fr.was studied under potculture conditions.The results showed that Suillus granulatus O.Kurttee and Suillusgrevillei Sing most probably belonged to one ectomycorrhizal fungus the same family,andthe rapid mycorrhizae formation produced strong stimulation on seeding in heightgrowth,radial growth and dry biomass accumulation,especially the increasing of drybiomass for the inoculated seeding was2-3times higher than that of the controlled ones.Drought stress was studied Pinus sylvestris var. mongolica Litv mycorrhizal seedlings（syn-thesized for130day）, with the purpose of ascertaining their drought resistance mechanism.The result indicated that mycorrhizul seedlings increased their drought resistance by man-ifold physiological and biochemical mechanisms that enhancing photosynthesis speed,augmenting the root top ratio, increasing the proline content and the activity of SOD andreducing the MDA．Among all the strains，Suillus granulatus O.Kurttee had the strongestdrought resistance capacity．Critical dead time of mycorrhizal seedlings was postponed61hours than the contrast.4. The mycorrhizal formation of tissue cultured plantletsThe effect of hormone on the callus induction and differentiation of mature embryoof Pinus sylvestris var. mongolica Litv was studied. And an efficiency regeneration Systemof the plant was estable.lished. The optimal conditions of each tissue culture stage wereestable.lished: The best proportion of medium of callus induction of Pinus sylvestris var. mongolica Litv are:1/2MS+2,4-D1.5mg·L^-1+6-BA0.5mg·L^-1The best proportion of mediu-m of the proliferated callus leaded of Pinus sylvestris var.mongolica Litv are:1/2MS+2,4-D1.0mg·L^-1+6-BA0.2mg·L^-1.The best proportion of medium of callus differentiation ofPinu-s sylvestris var. mongolica Litv are:1/2MS+IBA0.1mg·L^-1+6-BA1.0mg·L^-1;Thebestpropor-tion of medium of Adventitious buds elongation growththe of Pinus sylvestris var. mongol-ica Litv are:1/2MS+0.1%activated carbon; The best proportion of medium of rooting of Pi-nus sylvestris var. mongolica Litv are:1/2MS+NAA0.5mg·L^-1+IBA0.2mg·L^-1.After inocul-ated S.g and G-S.g for5months to the plantlets of Pinus sylvestris var. mongolica Litvafter transplanting, the mycorrhizal formation rates of S.g and G-S.g both were up to95%.The infection rates of S.g and G-S.g on the Pinus sylvestris var. mongolica Litv were up to75%. The heights,the ground diameters and the total dry weights of mycorrhizal plantletsof Pinus sylvestris var. mongolica Litv were Significantly improved than the contrast.All ofthe results manifested that after inoculated G-S.g, the growth of host plants were improveddefinitely.