| Quarantine Phoma host range are extensive,which includs cruciferae,leguminosae,solanaceae and other hundreds of species.It infects various parts of the host,leads to decline in crop production,lower quality and even death,and causes huge economic losses.This paper deeply studied the biological characteristics on 21 species of 12 quarantine Phoma and related isolates,such as Leptosphaeria maculans,Phoma pinodes,P.pinodella,P.glomerata.Focus on the study of Phoma spore activity staining and detection and identification of small molecular polypeptides of four quarantine representative species fungi :P.Pinodella,P.Glomerata,P.Terrestris,P.tracheiphila.Spores activity staining method of Phoma species and small molecular polypeptide detection method of quarantine Phoma were explored The results as follows:The biological characteristics on 21 isolates of 12 species research showes: the tested Phoma isolates had significant differences in colonial morphology、growing speeds and spores production.P.pinodella among isolates was quite different in colony morphology;The colonies were regular round,color is white,brown,red brown,part is light yellow,black and dark green,etc.The spores are single spore,colorless or light green,transparent,spherical,oval shaped or long rod-shaped,partly pear shaped.Spores of a few strains have one or two oil balls.Pycnidium spherical or near spherical shape,the outer walls is black or brown.Through the selection of the conditions of spore germination time,lethal temperature,type of reactive dyes,concentration,dyeing time and so on.Finally determine Phoma glomerata,Phoma terrestris 2 kinds of quarantine Phoma spores lethal temperature is 52℃,50 ℃.Based on the characteristics of PI can make the dead cells and live cells has a different fluorescence intensity under the excitation of laser light source at different wavelengths.Screening of working concentration 1mg/ml PI dye can clearly distinguish dead spores and live spore..The activity detection method of Phoma was established,and the repeatability of the method was verified.Five days PD liquid culture of P.glomerata,P.pinodella,P.terrestris,P.tracheiphila strain using homogenate +SDT lysis pyrolysis method to extract protein,iTRAQ analysis,check the library,identified four types of quarantine Phoma fungal differential protein.Ratio>2 and P <0.05 as the screening criteria,delete the predicted protein,specific expressed proteins number were 33,17,41,16 were selected.The significant difference ofprotein were the dipeptidyl-peptidase 5,50 S ribosomal protein L6,scytalone dehydratase,methylthio-ribulose-1-phosphate dehydratase,1,3,6,8-tetrahyd roxynaphthalene reductase in Phoma glomerata strain;The diaminopropionate ammonia-lyase,SURF-family protein Shy1,Ankyrin repeat and SAM domain containing protein 6,conidial yellow pigment biosynthesis polyketide synthase,3’-phosphoadenosine 5’-phosphatase isoform A,mycocerosic acid synthase in P.pinodella strain.The translationally-controlled tumor protein,ATP-dependent RNA helicase DBP5,acetyltransferase component of pyruvate dehydrogenase complex,probable Xaa-Pro aminopeptidase PEPP,delta(12)fatty acid desaturase in P.terrestris strain;The synaptic glycoprotein SC2,chitooligosaccha ride deacetylase,thiosulfate sulfurtransferase,Aldehyde dehydrogenase,Polyadenylate-binding protein 2(Poly(A)-binding protein 2)in P.tracheipila strain. |
