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DNA Barcoding of Fungi in the Forest Ecosystem of the Psunj and Papuk Mountains in Croatia

机译:克罗地亚Psunj森林生态系统中真菌的DNA条形码

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The saprotrophic, endophytic, and parasitic fungi were detected from the samples collected in the forest of the management unit East Psunj and Papuk Nature Park in Croatia. The disease symptoms, the morphology of fruiting bodies and fungal culture, and DNA barcoding were combined for determining the fungi at the genus or species level. DNA barcoding is a standardized and automated identification of species based on recognition of highly variable DNA sequences. DNA barcoding has a wide application in the diagnostic purpose of fungi in biological specimens. DNA samples for DNA barcoding were isolated from infected tree tissues, fungal fruiting bodies or fungal cultures. The ITS or ITS2 sequences of the fungal DNA were sequenced and aligned with the reference sequences in GenBank (NCBI) using BLAST. The size of ITS and ITS2 sequences were 512-584 bp and 248-326 bp, respectively. The sequences showed a high identity of 97.21%-100% at 98%-100% coverage with reference sequences in GenBank (NCBI). The exception was the species Amphilogia gyrosa that showed 95.65% identity at 100% coverage. Two fungi were determined at genus level: Cladosporium sp., and Cytospora sp., while 11 fungi were determined at species level: Alternaria alternata, Aureobasidium pullulans, Amphilogia gyrosa, Capronia pilosella, Cryphonectria parasitica, Exidia glandulosa, Epicoccum nigrum, Penicillium glabrum, Pezicula carpinea, Rosellinia corticium, and Stereum hirsutum.
机译:从管理单位东Psunj和克罗地亚Papuk自然公园的森林中收集的样品中检测到嗜肺繁殖,内生真菌和寄生真菌。疾病症状,结果体和真菌培养的形态,以及DNA条形码组合用于测定属或物种水平的真菌。 DNA条形码是基于对高度可变DNA序列的识别的物种的标准化和自动识别。 DNA条形码在生物标本中真菌的诊断目的具有广泛的应用。 DNA分段的DNA样品从感染的树组织,真菌果实或真菌培养物中分离出来。使用爆炸对真菌DNA的其或2个真菌DNA的序列与Genbank(NCBI)中的参考序列进行测序。其序列的大小分别为512-584 bp和248-326 bp。序列显示出97.21%-100%的高度为97.21%-100%,在Genbank(NCBI)中,参考序列覆盖率为98%-100%。例外是疗法疗法疗法,在100%覆盖范围内显示出95.65%的身份。在Genus水平中确定了两种真菌,Cladosporium sp。和Cytospora sp。,而11个真菌在物种等级确定:alertaria alternata,alurebasidium pullulans,疗法吡咯烷蛋白酶,葡萄球菌生成蛋白酶,低温植物寄生虫,exidia glandulosa,epicoccum nigrum,penicillium glabrum, Pezicula carpinea,rosellinia corticium和Stereum hirsutum。

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