Biological characters and rDNA ITS sequences of pathogen of poplar leaf blight

DNA was extracted from the strain of pathogen of poplar leaf blight using a modified CTAB method. ITS sequence （601bp） was initially amplified from the pathogen by using the universal primers ITSl and ITS4 （registered No, DQ011257）. Comparing to the nucleotide sequences acquired from GenBank database, the strain is clustered into the homogeneity with Alternaria alternate （AY787684） and Alternaria alternate （AY354228）, with a homology of 98%, thus the strain was checked as Alternaria alternata （Fr.） Keissler. The optimal conditions for conidia germination and mycelium growth of the pathogen were tested, The optimal temperature for conidia germinating and mycelium growth is 25℃, and the optimal pH value is 6. Mycelium grows rather slowly at 10℃ and 30℃ and growth stops at above 35 ℃. Among the six culture mediums tested, PDA ＋ poplar leaf juice medium is most favorable for mycelium growth.

Phylogeny of Aceraceae Based on ITS and trn L-F Data Sets

The nuclear encoded internal transcribed spacer (ITS) region and the plastid encoded trn L-F region were sequenced for 41 species of the Aceraceae, representing both genera Acer and Dipteronia, to reconstruct phylogeny of this family, especially within Acer. The analyses were performed in separate and combined sequence data sets, with the Sapindaceae and Hippocastanaceae being selected as outgroups. It was indicated that the Aceraceae was monophyletic and D. sinensis was basal to the rest of the family but the two genera of it might be not monophyletic because Dipteronia dyerana was nested within Acer. The result inferred from the combined data showed greater resolution within Acer than that from the two separate data sets. The monophyly of most sections in Xu's system (1996) were supported with high bootstrap values, and some relationships between (or among) sections were also inferred, such as sect. Palmata and sect. Microcarpa; sect. Platanoidea, sect. Lithocarpa and sect. Macrophylla; sect. Integrifolia, sect. Trifoliata and sect. Pentaphylla; and sect. Acer, sect. Goniocarpa and sect. Saccharina (sensu Ogata). However, the sectional status and circumscriptions of some of the above-mentioned sections should be further adjusted. It seemed that the Xu's delimitations of sect. Rubra and sect. Saccharodendran should be revaluated.

Investigation of Phylogenetic Relationship among Three Species of Channa Based on ITS Sequence Analysis

[ Objective ] This study aimed to analyze genetic variation of ribosomal ITS region sequences in Channa argus, C moculata and C. asiatica, and to in- vestigate the phylogenetic relationship among Charma species based on ITS sequences. [ Method] ITS sequences of three Channa species were amplified by PCR, cloned and assembled to obtain the full length of ITS sequences. [ Result] The full length of ITS sequences of C. argus, C. maculata and C. asiatica was 902, 927, and 902/903 bp, respectively. ITS sequences of C. argus, C. maculata and C. asiatica exhibited higher G ＋ C （72%） than A ＋ T. Interspecific nucleotide differences were significantly greater than intraspecific differences of these three Channa species. Thus, these remarkably differential ITS fragments could be used to identify C. argus, C. maculata and C. asiatica. Phylogenetic tree constructed by Neighbor-joining and Maximum Likehood methods showed that C. argus shared the lowest genetic distance with C. maculata and the highest genetic distance with C. asiatica. [ Conclusion] This study provided reference for classification, i- dentification, phylogenetic analysis and interspecific hybridization of Channa species.

DNA Molecular Identification of Botanical Origin in Chinese Herb Qingjiao

[Objective] The research aimed to distinguish Chinese herb Qingjiao from its botanical origin plants by comparing different DNA sequences,so as to provide a molecular basis for origin identification and quality evaluation.[Method] The cpDNA psbA-trnH and nrDNA ITS sequences of five Chinese herb Qingjiao plants,including Gentiana macrophylla pall.,Gentiana straminea Maxim.,Gentiana crassicaulis Duthie ex Burk.,Gentiana dahurica Fisch and Gentiana officinalis H.Smith,were amplified with PCR,and then sequenced by direct PCR sequencing method for homologous analysis.[Results] The length of cpDNA psbA-trnH of five plants was 316-318 bp;there were seven different haplotypes and seven variable sites;the GC content of the sequence was 21.2%;the phylogenetic clustering showed the same result as haplotype analysis.The length of nrDNA ITS sequence of five plants was 624-625 bp,there were five different haplotypes and 13 variable sites;the GC content of the sequence was 59.3%.The result of phylogenetic clustering suggested that G.dahurica and G.straminea,G.macrophylla and G.officinalis clustered together as sister clades,respectively.[Conclusion] The nucleotide differences of nrDNA ITS regions could be used for distinguishing botanical origin in Chinese herb Qingjiao.

Diversity and composition of culturable fungi in Horqin Sandy Land

Soil fungi play a key role in soil functional performance and ecological restoration.To understand the diversity and composition of culturable fungi in soils of Horqin Sandy Land,China,mobile dune,semi-fixed dune,fixed dune and sandy grassland were selected to investigate the soil fungal diversity using a traditional culture-dependent approach.ITS sequencing was applied to identify the fungal strains.The counts of culturable fungi increased significantly from mobile dune to sandy grassland along the gradient of sandy land restoration.The Shannon-Wiener,Simpson and Evenness indices of culturable fungi ranged from 1.26-1.71,0.22-0.37 and 0.83-0.87,respectively.A total of 27 fungal strains were isolated using dilution plate cultural technique.The 27 fungal isolates were clustered into three groups:Ascomycota,Basidiomycota and Mucoromycota at phylum level,indicating that Ascomycota was the dominant fungal phylum(88.9%of the total).The isolated fungi were grouped into 3 phyla,5 classes,6 orders,11 families and 13 genera.The results show that culturable fungi were diverse in sandy land soils and fungal isolates have potential function in lipid turnover,cellulose degradation and ethanol,glucose and fatty acid production.Future studies should be carried out to explore their ecological and biological function for degraded sandy land restoration.

Morphology and Molecular Identification of Ulva Forming Green Tides in Qingdao,China

Green tides are caused by the proliferation of chlorophytes under suitable hydrographic conditions.These blooms lead to environmental degradation and negatively impact the waters and seagrass beds,as well as fishing and other recreational activities in the bay.A comprehensive ecological understanding of the bloom dynamics,including the origin and persistence,is needed to foster management decisions.The algae in the great majority of green tide blooms usually belong to two genera of Ulvophyceae,Ulva and Enteromorpha.Ulva has been observed more often in recent years.In China,green tides occurred for the first time in the middle area of the Yellow Sea in 2007,and a large-scale algae blooming broke out in the middle and southern areas of the Yellow Sea in late May 2008.We identified them as Ulva prolifera by comparative analysis of the rDNA internal transcribed spacer 1 (ITS1),5.8S and ITS2 sequences in combination with microscopic observation.Morphological differences were found between the free-floating algae and the attached thalli.Various reproduction patterns of the free-floating algae include sexual,asexual and vegetative propagations,which played important roles in the long-term green tide persistence in China.The ITS sequences of the blooming algae were identical to those of the samples from the Lianyungang sea area but were different from the attached samples from the Qingdao sea area.The results infer that the blooms are originated from other sea areas rather than from the local attached populations.