MOEKETSI E KHOLO
Ho matlafala ha temo ho fokotsa ho rarahana ha marang-rang a likokoana-hloko le bongata ba lejoe la bohlokoa la lekhetho metsong.
Sequencing ea Amplicon e bolelele bo felletseng (ITS) |PacBio HIFI
Lintlha-khōlō
Aho matlafala ha temo ho se ho ntse ho e-na le mathata a mangata ka lebaka la liphello tse mpe tsa tikoloho tse kenyeletsang ts'ebeliso e fokolang ea limatlafatsi, metsi a tsoang ka tlas'a metsi, ho senyeha ha boleng ba mobu, joalo-joalo. Mekhoa e meng ea temo e kenyelletsang ho se phethoe le temo ea manyolo e amohetsoe ka bophara ho fokotsa kotsi.Sechaba sa likokoana-hloko se bapala karolo ea bohlokoa tlhahisong le ts'ebetsong ea agroecosystem.Leha ho le joalo, ha ho hlake hore na mekhoa e fapaneng ea temo e ama microbiota joang.
Moqapi oa Teko
Liteko
Slisampole tsa oli le motso (DNA) li ne li tsoa masimong a koro ho tsoa masimong a 60 a temo (20 ka 'ngoe)
Gho roka: 1. Kopano (ka ho lema mobu);2. Tumellano (ha ho na mobu);3. Mapolasi a tlhaho
Sleano la equencing: Full-length amplicon sequencing (ITS)
Primers: ITS1F-ITS4 (e shebane le sebaka sohle sa ITS ~630 bp)
Ssethaleng sa equencing: PacBio RS II
Tlhahlobo ea Bioinformatics
Liphetho
Oka karolelano ea li-OTU tse 357 li ile tsa khetholloa sebakeng se seng le se seng le kakaretso ea li-OTU tse 837 tsa libaka tsohle tse 60.Mefuta e fapaneng ea alpha ea metse ea fungal ea metso e ne e sa bontše phapang e kholo har'a mekhoa e meraro ea temo.Leha ho le joalo, lihlopha tse tharo tse ikhethileng li ile tsa thehoa tlhahlobisong ea mefuta-futa ea beta, e bontšang liphello tse matla tsa tsamaiso ea temo mohahong oa sechaba sa fungal.
Setšoantšo sa 1. Phapang ea alpha (index ea Shannon le sebopeho sa sechaba) le tlhahlobo ea mefuta-futa ea beta (tlhahlobo ea li-canonical tsa likhokahano tse kholo) ho sechaba sa fungal.
TLejoe la bohlokoa la lekhetho le ile la hlalosoa ho ipapisitsoe le marang-rang a akaretsang a sechaba sa fungal ho pholletsa le tsamaiso e meraro ea temo: li-node tse 10 tse nang le tekanyo e phahameng ka ho fetisisa, kamano e haufi-ufi e phahameng ka ho fetisisa le bohareng bo tlaase ka ho fetisisa bo ile ba khethoa.Tse supileng tsa tsona e ne e le tsa litaelo tsa mycorrhizal.
Setšoantšo sa 2. Marang-rang ka kakaretso holim'a metse e meholo ea fungal ea mekhoa e meraro ea temo
Fmarang-rang a ikhethileng a sistimi e bonts'itseng khokahanyo e phahameng haholo ho marang-rang a tlhaho a nang le likarolo tse fetang habeli le li-node tse hoketsoeng ho feta marang-rang a no-till le a tloaelehileng.Ho feta moo, marang-rang a temo ea manyolo a ne a na le lejoe la bohlokoa la lekhetho (taemane) ha le bapisoa le tse ling kaofela, tse neng li tšehetsa ho rarahana le khokahanyo ea eona.
Setšoantšo sa 3. Metsotso ea fungal ea tsamaiso ea temo
Akamano e matla e mpe lipakeng tsa matla a temo le khokahano ea marang-rang ea fungal e ile ea bonoa.Tlhahlobo ea meru e sa reroang e senotse bakhanni ba ka sehloohong ba lekhetho la lejoe la bohlokoa: phosphorus ea mobu, bongata bo bongata, pH le mycorrhizal colonization.
Setšoantšo sa 4. Matla a temo le khokahanyo ea marang-rang ho pholletsa le mekhoa e meraro ea temo (A le B);Tlhahlobo ea meru e sa reroang(C) le Kamano lipakeng tsa matla a temo le bokolone ba AMF (D)
Thekenoloji
Tatelano ea amplicon ea bolelele bo felletseng
As "Tatelano ea Moloko oa Boraro" e tlang sethaleng, mefokolo ea libaka tse lebisitsoeng le mathata a kopano ea de novo e hlōtsoe.Pacific Bioscience (PacBio) e atlehile ho atolosa ho baloa ha tatellano ho li-kilobase tse mashome, tse re lumellang ho fumana lipalo tse feletseng tsa 16s rRNA (1,000 bp-1,500 bp) ka libaktheria kapa 18S rRNA (1,500 bp-2,000 bp) le ITS libaka (400 bp-900 bp) ka eukaryotic.Pono e pharalletseng ea sebaka sa liphatsa tsa lefutso e matlafalitse haholo tharollo ea litlhaloso tsa mefuta le liphatsa tsa lefutso tse sebetsang.Taba e telele e amehileng mabapi le ho nepahala ha motheo e rarollotsoe ke PacBio CCS ho itokisa, e hlahisang HIFI e baloang ka ho nepahala ho fetang 99% ho bala.
Ts'ebetso ho tlhaloso ea OTU
Tho fumana melemo ea ho bala nako e telele le ho hlahisa lintlha tse phahameng, ho nepahala ha litlhaloso ho ka eketsoa ka mokhoa o tsotehang le ho finyella qeto ea "maemo a mefuta" ho khethollang likokoana-hloko.
Referense
Banerjee, Samiran, et al."Ho matlafala ha temo ho fokotsa ho rarahana ha marang-rang a likokoana-hloko le bongata ba lejoe la bohlokoa la lekhetho metsong."Koranta ea ISME (2019).
Theknoloji le Lintlha-khōlō e ikemiselitse ho arolelana ts'ebeliso ea morao-rao e atlehileng ea mekhoa e fapaneng ea tatellano ea maemo a holimo mabaleng a fapaneng a liphuputso hammoho le mehopolo e bohlale moralong oa liteko le ho rafa data.
Nako ea poso: Jan-08-2022