NKE MICROBIAL ECOLOGY
Agricultural intensification na-ebelata mgbagwoju anya microbial netwọk na ụba nke keystone taxa na mgbọrọgwụ
Usoro Amplicon zuru oke (ITS) |PacBio HIFI
Isi ihe
AMmụba ọrụ ugbo abụrụla nke na-enwewanye nsogbu n'ihi mmetụta ọjọọ ya na gburugburu ebe obibi gụnyere adịghị mma iji nri eme ihe na-adịghị mma, eutrophiation nke goundwater, mmebi nke àgwà ala, wdg. Usoro ọrụ ugbo ọzọ gụnyere enweghị-till na organic ugbo ka anabatara nke ọma iji belata mmebi ahụ.Obodo Microbial na-arụ ọrụ dị mkpa na nrụpụta na nkwado nke agroecosystem.Agbanyeghị, ọ bụ ihe edoghị anya na ka usoro ọrụ ugbo dị iche iche si emetụta microbiota mgbọrọgwụ.
Nleba anya
Nnwale
SIhe nlele mmanụ na mgbọrọgwụ (DNA) sitere n'ubi ọka wit sitere na ala ugbo iri isii (20 nke ọ bụla)
Grouping: 1. Mgbakọ (na tillage);2. Mgbakọ (enweghị-ịgba osisi);3. Organic ugbo
SAtụmatụ equencing: usoro amplicon ogologo zuru oke (ITS)
PRimers: ITS1F-ITS4 (na-eche na mpaghara ITS dum ~ 630 bp)
Sequencing ikpo okwu: PacBio RS II
Nyocha bioinformatic
Nsonaazụ
On nkezi nke 357 OTU ka achọpụtara n'otu saịtị yana ngụkọta nke 837 OTU nke saịtị 60 niile.Alfa dị iche iche nke obodo fungal mgbọrọgwụ egosighi nnukwu ọdịiche n'etiti usoro ọrụ ugbo atọ.Otú ọ dị, e hiwere ụyọkọ atọ dị iche iche na nyocha ụdị dị iche iche beta, na-egosi mmetụta siri ike nke usoro ọrụ ugbo na nhazi obodo fungal.
Ọgụgụ 1. Alfa diversity (Shannon index na obodo mejupụtara) na beta diversity analysis (canonical analysis of principal coordinates) na mgbọrọgwụ fungal obodo.
TA kọwapụtara taxa keystone dabere na netwọkụ nke obodo fungal gafee usoro ọrụ ugbo atọ: ọnụ ọnụ iri kacha elu nwere ogo kachasị elu, etiti nso kacha nso na etiti etiti kacha ala ka ahọpụtara.Asaa n'ime ha bụ nke mycorrhizal iwu.
Ọgụgụ 2. N'ozuzu netwọk na mgbọrọgwụ fungal obodo nke atọ ugbo usoro
FNetwọk ndị a kapịrị ọnụ nke ngwa agha gosipụtara njikọ dị elu nke ukwuu na netwọkụ organic nwere ọnụ ọnụ ugboro abụọ yana ọnụ ọnụ karịa karịa netwọk enweghị-ruo na nke mbụ.Ọzọkwa, netwọk ọrụ ugbo organic nwere ọtụtụ keystone taxa(diamond) ma e jiri ya tụnyere ndị ọzọ, nke kwadoro mgbagwoju anya na njikọta ya.
Ọgụgụ 3. Usoro ọrụ ugbo-kpọmkwem mgbọrọgwụ fungal netwọk
AAhụrụ njikọ ọjọọ siri ike n'etiti ike ọrụ ugbo na njikọ netwọk fungal mgbọrọgwụ.Nyocha nke oke ọhịa na-ekpughere isi ihe na-akpata taxa keystone: phosphorus ala, njupụta buru ibu, pH na mycorrhizal colonization.
Ọgụgụ 4. Ịrụ ọrụ ugbo na njikọ netwọk n'ofe usoro ugbo atọ (A na B);Nyocha oke ọhịa (C) na mmekọrịta dị n'etiti ike ọrụ ugbo na nchịkwa AMF (D)
Nkà na ụzụ
Usoro amplicon ogologo zuru oke
As "Sequencing ọgbọ nke atọ" na-abịa na ogbo, emeri adịghị ike na mpaghara ezubere iche na nsogbu na de novo mgbakọ.Pacific Bioscience (PacBio) ejirila nke ọma gbasaa ịgụ usoro na iri puku kilobases, nke na-enye anyị ohere inweta ọgụgụ zuru oke nke 16s rRNA (1,000 bp-1,500 bp) na nje bacteria ma ọ bụ 18S rRNA (1,500 bp-2,000 bp) na ITS mpaghara (400 bp-900 bp) na eukaryotics.Echiche sara mbara nke mkpụrụ ndụ ihe nketa mere ka mkpebi nke nkọwapụta ụdịdị na mkpụrụ ndụ ihe nketa na-arụ ọrụ dịkwuo mma.PacBio CCS edozila okwu a na-eche ogologo oge maka izi ezi nke ntọala, nke na-emepụta HIFI na-agụ ihe karịrị 99% ziri ezi.
Arụmọrụ na nkọwa OTU
TN'ịchọta uru nke ma ogologo ọgụgụ na nke dị elu, izi ezi nke nkọwa nwere ike ịba ụba nke ukwuu ma nweta mkpebi "ụdị ọkwa" na njirimara microbial.
Ntụaka
Banerjee, Samiran, et al."Mbawanye ọrụ ugbo na-ebelata mgbagwoju anya netwọk microbial na ụbara taxa keystone na mgbọrọgwụ."Akwụkwọ akụkọ ISME (2019).
Teknụzụ na isi ihe bu n'obi kesaa ngwa kacha aga nke ọma na nso nso a nke teknụzụ usoro usoro dị elu dị iche iche na mpaghara nyocha dị iche iche yana echiche mara mma na nhazi nnwale na ngwuputa data.
Oge nzipu: Jan-08-2022