PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) analysis of cpDNA and PCR-RFLP of rDNA were performed to assess the phylogenetic relationship in seven species of the family Cornaceae. Four pairs of primers, trnD/trnT, trnH/trnK, psaA/trnS and rbcL/ORF106, and one restriction enzyme (HinfI) were used for RFLP-PCR of cpDNA, and two primer pairs ITS5/ITS2 and ITS3/ITS4 with one restriction enzyme (HhaI) were used for ITS1 and ITS2 regions of rDNA, successively. Restriction products produced by trnH/trnK-HinfI and psaA/trnS-HinfI for cpDNA and ITS5/ITS2-HhaI and ITS3/ITS4-HhaI for rDNA, generated very informative bands from all of the seven species studied. A total of 147 scores were recorded and used in PHYLIP (phylogeny inference package) to generate a combined phylogenetic tree. The family was divided into two clades: the Aucuba clade and the cornaceous clade. The cornaceous clade further divided into two subclades: the previously suggested blue-or white-fruited dogwoods (C. walteri and C. macrophylla) and the Cornelian cherries (C. officinalis), and the big-bracted dogwoods (C. kousa and C. florida). The opposite-leaved blue-or white-fruited dogwoods were found to be close sisters to the alternate-leaved blue-or white-fruited dogwoods and all sisters to the Cornelian cherries. C. kousa and C. florida were found to be close sisters under big-bracted dogwoods. Based on the topology portrayed by the phylogenetic tree obtained, it is possible to conclude that big-bracted dogwoods are the most primitive member followed by the intermediates -the cornelian cherries, and the blue-or white-fruited dogwoods are the most advanced members, while the Aucuba clade is the most primitive in the family.
Microbes in forest are very important due to not only to enhance soil fertility but also maintain a healthy ecosystem by supplying the energy available to living organisms by producing various kinds of enzymes related to degradation of lignocellulosic biomass. In order to isolate a lignocellulosic biomass degrading bacterial strain from the Jurassic park located in Gyeongnam National University of Science and Technology, We used the Luria-Bertani-Carboxymethyl cellulose (CMC) agar trypan blue method containing 0.4 % carboxymethyl cellulose and 0.01 % trypan blue. As a result, we isolated a bacterial strain showing both activity on the CMC and xylan. To identify the isolated strain, 16S rRNA sequencing and API kit analysis were used. The isolated strain turned out to belong to Bacillus species and then named Bacillus sp. GJY. In the CMC zymogram analysis, it showed that one active band of about 28kDa in size is present.
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