● Ho itšehla thajana le ho lema ntle ho tefo bakeng sa profiling ea sechaba sa likokoana-hloko
● Qeto e phahameng ea ho fumana mefuta e mengata e fokolang lisampoleng tsa tikoloho
● Khopolo ea "meta-" e kopanya likarolo tsohle tsa likokoana-hloko boemong ba ts'ebetso, boemo ba mefuta le liphatsa tsa lefutso, tse bontšang pono e matla e haufi le 'nete.
● BMK e bokella boiphihlelo bo bongata mefuteng e fapaneng ea lisampole ka lisampole tse fetang 10,000 tse sebetsitsoeng.
Sethala | Tatelano | Lintlha tse khothalelitsoeng | Nako ea ho fetoha |
Illumina NovaSeq 6000 | PE150 | 6 G/10 G/20 G | 45 Matsatsi a ho sebetsa |
● Taolo ea boleng ba data e tala
● Kopano ea Metagenome
● Liphatsa tsa lefutso tse sa hlokahaleng le tlhaloso
● Tlhahlobo ea mefuta-futa ea mefuta
● Tlhahlobo ea ho se tšoane ha liphatsa tsa lefutso
● Tlhahlobo ea lihlopha
● Tlhahlobo ea mokhatlo khahlanong le lintlha tsa liteko
Bakeng saLintlha tsa DNA:
Mofuta oa Mohlala | Chelete | Ho tsepamisa maikutlo | Bohloeki |
Lintlha tsa DNA | > 100 ng | > 1 ng/μl | OD260/280= 1.6-2.5 |
Bakeng sa mehlala ea tikoloho:
Mofuta oa mohlala | Mokhoa o khothaletsoang oa sampole |
Mobu | Chelete ea mohlala: hoo e ka bang.5 g;Lintho tse setseng tse omeletseng li hloka ho tlosoa holimo;Sila likotoana tse kholo 'me u fete ka har'a filthara ea 2 mm;Lisampole tsa Aliquot ka har'a EP-tube kapa cyrotube bakeng sa ho behelloa. |
Mantle | Chelete ea mohlala: hoo e ka bang.5 g;Bokella le lisampole tsa aliquot ho EP-tube kapa cryotube e sa sebetseng bakeng sa ho behelloa. |
Likahare tsa mala | Mehlala e hloka ho sebetsoa tlas'a boemo ba aseptic.Hlatsoa lithane tse bokeletsoeng ka PBS;Hlakola PBS 'me u bokelle precipitant ka EP-tubes. |
Sludge | Chelete ea mohlala: hoo e ka bang.5 g;Bokella le aliquot sludge sampole ho EP-tube e hloekisitsoeng kapa cryotube bakeng sa ho behelloa |
Metsi | Bakeng sa sampole e nang le palo e lekanyelitsoeng ea likokoana-hloko, joalo ka metsi a pompo, metsi a seliba, joalo-joalo, Bokella bonyane metsi a 1 L 'me u fete ka 0.22 μm filthara ho ntlafatsa likokoana-hloko tse holim'a lera.Boloka lera ka har'a tube e hloekisitsoeng. |
Letlalo | Hlakola bokaholimo ba letlalo ka hloko ka swab e hloekisitsoeng ea k'hothone kapa lehare la ho buoa 'me u le kenye ka har'a tube e sa hloekang. |
Emisa lisampole ka har'a naetrojene e metsi bakeng sa lihora tse 3-4 'me u li boloke ka har'a metsi a naetrojene kapa -80 degree ho boloka nako e telele.Ho hlokahala ho romelloa ka sekepe se nang le leqhoa le omileng.
1.Histogram: Kabo ea mefuta
2.Liphatsa tsa lefutso tse sebetsang tse hlalositsoeng ho KEGG metabolic pathways
'Mapa oa 3.Heat: Mesebetsi e fapaneng e itšetlehileng ka bongata ba liphatsa tsa lefutso4.Li-Circos tsa liphatsa tsa lefutso tsa CARD tsa lithibela-mafu
Taba ea BMK
Ho ata ha liphatsa tsa lefutso tsa lithibela-mafu le likokoana-hloko tsa baktheria hammoho le motso oa mobu oa mangrove
E hatisitsoe:Journal of Hazardous Materials, 2021
Leano la tatellano:
lintho tse bonahalang: Likaroloana tsa DNA tsa likaroloana tse 'nè tsa lisampole tse amanang le motso oa mangrove: mobu o sa lengoang, rhizosphere, episphere le endosphere compartments.
Sethala: Illumina HiSeq 2500
Sepheo: Metagenome
16S rRNA mofuta V3-V4 sebakeng
Liphetho tsa bohlokoa
Metagenomic sequencing le metabarcoding profiling mabapi le ho tsoela pele ka metso ea mobu oa lifate tsa mangrove li ile tsa sebetsoa molemong oa ho ithuta ka phatlalatso ea liphatsa tsa lefutso tsa lithibela-mafu (ARGs) ho tloha mobung ho ea ho limela.Lintlha tsa Metagenomic li senotse hore 91.4% ea liphatsa tsa lefutso tsa lithibela-mafu li ne li atisa ho tsejoa likarolong tsohle tse 'nè tsa mobu tse boletsoeng ka holimo, tse bontšang mokhoa o tsoelang pele.16S rRNA amplicon sequencing e hlahisitse tatelano ea 29,285, e emelang mefuta e 346.Ho kopanngoa le profiling ea mefuta ka tatellano ea li-amplicon, phatlalatso ena e fumanoe e ikemetse ho microbiota e amanang le metso, leha ho le joalo, e ka nolofalloa ke mobile ea likarolo tsa lefutso.Boithuto bona bo hloaile phallo ea li-ARG le likokoana-hloko tse tsoang mobung ho ea limeleng ka ho tsoela pele ho hokahantsoeng ha metso ea mobu.
Referense
Wang, C. , Hu, R. , Strong, PJ , Zhuang, W. , & Shu, L. .(2020).Ho ata ha liphatsa tsa lefutso tsa lithibela-mafu le likokoana-hloko tsa baktheria ho latela metso ea mobu oa mangrove.Journal of Hazardous Materials, 408, 124985.