Study On The Molecular Phylogenetic Relationships Of Alternaria Isolated From Jujube In South Of Xinjiang

The photo-thermal resources is rich in Xinjiang, the prominent character of Xinjiang is the drought andthe precipitation is always very little, it’s suitable for the jujube growth,jujube area have been more than0.4million hm2,but since2010, the jujube fruits and jujube leaf disease is more and more serious,it causegreat economic losses.Confusion and uncertainty of jujube fruit and jujube leaf disease pathogen speciesbring the difficult to control. This paper collecte typical samples with the jujube fruits and jujube leafdisease from the mainly produced jujube district in Southern Xinjiang.We isolated,purified and preservedthe jujube fruits and jujube leaf disease pathogen,after pathogenic test,morphological identification and ITS,EF-1α,β-tubulin and ATPase gene sequence analyze methods were combined to identify the main pathogenfor the jujube fruits and jujube leaf disease.That through the experiment test RAPD and ISSR analysis iswhether can as an additional method to species identification. The results are as below.1.From sample1and sample2,we obtained three kinds of isolates respectively and two kinds fromsample3.we just got nine kinds of fungal isolates from the4th to the12th sample (every sample got onekind).By Koch’s method testing sample1-3, we found that HT224-2, HTKS-3, HTYT-2are the pathogensbacteria.2.By referring to the method of Zhang Tianyu, according to conidial shape, size range, septation,thesporulation phenotype, presence or absence of beak and morphological characteristics, in combination withthe ITS sequence, EF-1α,β-tubulin and ATPase gene sequence analyse,HTKS-3and HTYT-2wereidentified as Alternaria alternata（Fr.：Fr.）Keissler.3.The optimal ISSR and RAPD reaction system for Alternaria small-spored species were established:theconcentration of Mg2+, dNTPs, primer, Taq polymerase were2.00mmol/L,0.15mmol/L,0.60μmol/L,1.25U in the25μl RAPD reaction system; the concentration of Mg2+, dNTPs, primer, Taq polymerase were2.50mmol/L,0.10mmol/L,0.30μmol/L,1.25U in the25μl ISSR reaction system.4.By useing cluster analysis for RAPD polymorphic marker of13kinds of Alternaria small-sporedspecies, it found that12kinds of Alternaria alternata were clustered together, Alternaria tenuissima wereclustered alone when genetic similarity indexes was0.61, it shows that RAPD analysis can as an additionalmethod to species identification; By useing cluster analysis for ISSR polymorphic marker of13kinds ofAlternaria small-spored species, ISSR clustering results showed that12kinds of Alternaria alternata wereclustered together, Alternaria tenuissima were clustered alone when genetic similarity indexes was0.71, itshows that ISSR analysis can as an additional method to species identification.