Isolation And Biochemical Composition Determination Of Ten Endophytic Algae Of Lichen And Study On The Algae Crust Condition

In this study,five lichens species were used to isolate the endophytic algae of lichens witch were distributed in Nanshan Bayi Forest Farm in Xinjiang.based on the morphological character and rDNA-ITS,18S-rRNA,16S-rRNA sequencing of endophytic algae of lichens,identified its taxonomic status and nutrient contents.The endophytic algae of lichens with high nutrient were screened out for optimizing its growth condition,and studied the formation of biological crust conditions of two photobiont cyanobacterias.The results are as follows:1.10 endophytic algae of lichens were isolated from 5 lichens species by homogenization coating method and isolated three endophytic algae of lichens(I1,P.E7,K)from Platigera elisabethae Gyeln.two algae(C,L1)were isolated from the ground Peltigera rufescens(Weis.)Humb.algae(P.A)were isolated from Peltigera apthosa(L.)Willd.Two endophytic algae of lichens(D,H1)were isolated from Peltigera membranacea(Ach.)Nyl.two lichens algae(P.E1,P.M2)were isolated from Peltigera canina(L).Willd.2.the classification status of endophytic algae of lichens was determined by morphological characteristics and rDNA-ITS,18S-rRNA and 16S-rRNA sequencing.The sequencing results of 3 median sites were consistent,among which four were algae cyanobacteria and six were greens algae.endophytic algae of lichens I1,H1belong to Microcoleus vaginatus and algae beads P.E1,L1 belong to Nostoc.Algae C belongs to Dictyosphearium,Algae D belongs to Trebouxiophyte,Algae K and P.A are closely related to Stichococcus bacillaris,Algae beads P.M2 belongs to the genus Coccomyxa.3.In this study,the soluble protein,polysaccharide,pigment and 20 amino acid contents of 10 endophytic algae of lichens were determined by Camas brilliant blue Staining method,Beryllium copper reagent method,Acetone reagent method and Gas chromatography.The results showed that the content of soluble protein and soluble sugar was the highest in(Trebouxiophyte)D,and the content values were331.9?g/mg,3268.1?g/mg respectively.The content of chlorophyll b reached 1387.6mg/L;the content of chlorophyll a and carotenoid was the highest in algae beads C(Dictyosphearium)and D(Trebouxiophyte),the content values were 1566.9 mg/L and1410.7 mg/L,1387.6 mg/L,1042.1 mg/L respectively.The results of amino acid content determination showed that the essential amino acids were in high level in the algae beads C,D and K,accounting for 36.70%,37.78%and 21.64%respectively.The percentage of total non-essential amino acids is:57.97%,60.71%,27.62%.4.Studied the effects of five different pH conditions,carbon source,carbon source concentration,nitrogen source and nitrogen source concentration on the growth of three endophytic algae of lichens(C,D,K).The results showed that pH=7,glacial acetic acid as carbon source,concentration of 1%,glycine as nitrogen source,concentration of 1%is most suitable for growth of endophytic algae of lichens.5.Two kinds of symbiotic cyanobacteria Microcoleus vaginatus and Nostoc,were used as research materials to study the forming artificial soil microbiotic algae crust conditions of Symbiotic cyanobacteria under laboratory simulation.The results showed that The 10%sand content,the 3:1(Microcoleus vaginatus:Nostoc sp.)inoculation amount and the10?g/cm~2 algae inoculation amount is suitable for forming artificial soil microbiotic algae crust.