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Wyświetlanie 1-2 z 2
Tytuł:
Zastosowanie regionu ITS1/2 rDNA i 18S rDNA do badania mykobioty gleby leśnej
Use of ITS1/2 rDNA and 18S rDNA in studies of the forest soil mycobiota
Autorzy:
Behnke-Borowczyk, J.
Kwaśna, H.
Powiązania:
https://bibliotekanauki.pl/articles/989532.pdf
Data publikacji:
2016
Wydawca:
Polskie Towarzystwo Leśne
Tematy:
lesnictwo
gleby lesne
mikroorganizmy glebowe
grzyby glebowe
struktura zbiorowisk
metody badan
metody molekularne
DNA rybosomalny
region ITS1/2
region 18S
detection
forest
its1/2 rdna
ns1
ns2
18s rdna
microorganisms
mycobiota
soil
Opis:
The aim of the studies was to check the usefulness of ITS1/2 rDNA and 18S rDNA regions in the molecular investigation of forest soil microbiota structure. Soil studied, originated from a 1−year−old plantation and a 40−year old stand of Scots pine located in Bierzwnik and Międzychów forest districts located 200 km apart. The hypothesis assumed that both approaches lead to the discovery of abundant microbiota communities with different structures and with rare common species. The environmental DNA was extracted with a Power Soil ® DNA Isolation Kit from two soil samples in each site. The ITS1/2 rDNA was amplified with specific primers ITS1 and ewfitsrev 1, and 18S rDNA with universal primers NS1 and NS2. PCR products were cloned into pGEM−T Easy. Inserts were primarily selected in blue/white screening on a X−gal medium. Representative clones were further selected in two separate RFLP analyses with HhaI and BsuRI restriction enzymes. Representative clones purified and sequenced using the Sanger Method in the DNA Research Centre (Poznań). Each sequence was identified to the lowest taxonomic rank. Ninety to 233 clones with DNA of 5−44 taxa including 3−37 taxa of fungi were obtained from 4 samples of soil. After application of ITS1/2 rDNA and 18S rDNA, the fungal DNA was detected respectively in 89,60−100,00% and 11,77−64,8% clones and the number of fungal species detected was respectively 12−37 and 3−19. Fungi were represented by four orders: Chytridiomycota, Zygomycota, Ascomycota and Basidiomycota. Both primers also amplified also DNA of other organisms (mostly from Animalia and Protista Kingdom) represented by 0−9 taxa. If compared, the application of forest soil microbiota structure with ITS1/2 rDNA and 18S rDNA led to detect a lower abundance of fungi and a bigger abundance of other organisms. Considering the higher number of clones and taxa recognized, the region of ITS1/2 rDNA was more effective in the studies of the soil microbiota structure. The region of 18S rDNA was efficient in local detection of Chytridiomycota and Zygomycota and of rare species of fungi from Ascomycota and Basidiomycota. Despite the deficiency of NCBI database the use of the 18S rDNA region in studies on fungal community the region should be included in molecular studies of fungal diversity. It is concluded that studies on the biodiversity of soil microorganisms need the application of a few independent methods of detection and identification.
Źródło:
Sylwan; 2016, 160, 07; 564-572
0039-7660
Pojawia się w:
Sylwan
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Wpływ metodyki badań na ocenę struktury zbiorowisk mikroorganizmów w glebie leśnej
Effect of the methodology of studies on the structure of the microorganisms communities in the forest soil
Autorzy:
Behnke-Borowczyk, J.
Kwaśna, H.
Powiązania:
https://bibliotekanauki.pl/articles/989286.pdf
Data publikacji:
2016
Wydawca:
Polskie Towarzystwo Leśne
Tematy:
lesnictwo
gleby lesne
mikroorganizmy glebowe
grzyby mikroskopowe
struktura zbiorowisk
metodyka badan
metody klasyczne
metody molekularne
startery NS1
startery NS2
DNA
region 18S
classical method of isolation
fungi
microorganisms
ns1
ns2
18s rdna
soil
Opis:
Two different communities of microorganisms were identified in soils by application of the classical method of fungi isolation (soil dilution, culturing on artificial media, morphotyping) and a molecular method (extraction of the environmental DNA, amplification with universal primers NS1 and NS2, cloning and sequencing of representative clones). No organisms were common to both communities. Apart from rare representatives of the Animalia, communities included single fungus−like Eucarya belonging to the Protista, Class Oomycota, and numerous fungi belonging to Chytridiomycota, Zygomycota, Ascomycota and Basidiomycota orders. In total, 88 species were identified in four soil samples. Fungi were mostly Ascomycota. The classical method was particularly effective in detection of fungi important for creation of phytosanitary conditions of soil, i.e. antagonists (Penicillium, Tolypocladium and Trichoderma) and potential stimulants (dark−pigmented Hormiactis candida, Humicola spp. and Phialophora spp.) of phytopathogens (including the common forest genera Armillaria and Heterobasidion). Application of the classical method allowed the detection of mycorrhizal Ascomycota from the genus Oidiodendron. Application of the molecular method allowed the detection of 13 mycorrhizal Basidiomycota. Although primers NS1 and NS2 were designed from a match with DNA of culturable organisms, they also amplified the DNA of non−culturable organisms. This emphasizes their potential usefulness in studies of the biodiversity of microorganisms in environmental samples. The shortage of reference sequences in the database discourages use of the 18S rDNA region in studies on fungal communities. The studies on the biodiversity of microorganisms need the application of a few independent methods of detection and identification.
Źródło:
Sylwan; 2016, 160, 06; 492-503
0039-7660
Pojawia się w:
Sylwan
Dostawca treści:
Biblioteka Nauki
Artykuł
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