ØUkwahlukaniswa kunye nokulinywa simahla kwiprofayili yoluntu lwemicrobial
ØIsisombululo esiphezulu ekufumaneni iintlobo eziphantsi kobuninzi kwiisampuli zokusingqongileyo
ØIngcamango ye "meta-" idibanisa zonke iimpawu zebhayoloji kwinqanaba lokusebenza, inqanaba leentlobo kunye nenqanaba lemfuza, ebonisa umbono oguqukayo osondeleyo kwinyani.
ØI-BMK iqokelela amava amakhulu kwiintlobo zeesampulu ezahlukeneyo ezineisampulu ezingaphezulu kwe-10,000 ezicutshungulweyo.
UkulandelelanisaIqonga | Ithala leencwadi | Imveliso yedatha ecetyiswayo | Ixesha eliqikelelweyo lokujika |
Illumina NovaSeq 6000 | PE250 | 50K/100K/300K iithegi | Iintsuku ezingama-30 |
üUlawulo lomgangatho wedatha ekrwada
üIndibano yeMetagenome
üIseti yemfuza engafunekiyo kunye nenkcazo
üUhlalutyo lweentlobo ezahlukeneyo zeentlobo
üUhlalutyo lokwahluka komsebenzi wemfuza
üUhlalutyo lwamaqela
üUhlalutyo lombutho ngokuchasene nezinto zovavanyo
KubaIzicatshulwa zeDNA:
Uhlobo lweSampuli | Isixa | Ukugxila | Ubunyulu |
Izicatshulwa zeDNA | > 30 ng | > 1 ng/μl | OD260/280= 1.6-2.5 |
Ngeesampulu zokusingqongileyo:
Uhlobo lwesampuli | Inkqubo yesampulu ecetyiswayo |
Umhlaba | Isixa sesampulu: malunga.5 g;Into eseleyo ebunayo kufuneka isuswe kumphezulu;Gaya iinqununu ezinkulu kwaye udlule kwi-2 mm filter;Iisampulu ze-Aliquot kwityhubhu ye-EP okanye i-cyrotube yogcino. |
Ilindle | Isixa sesampulu: malunga.5 g;Qokelela kwaye iisampulu ze-aliquot kwi-EP-tube engcolileyo okanye i-cryotube yokubhukisha. |
Iziqulatho zamathumbu | Iisampuli kufuneka ziqwalaselwe phantsi kwemeko ye-aseptic.Hlamba izicubu eziqokelelweyo nge-PBS;I-Centrifuge i-PBS kwaye uqokelele i-precipitant kwi-EP-tubes. |
Udaka | Isixa sesampulu: malunga.5 g;Qokelela kwaye isampuli ye-aliquot yodaka kwi-EP-tube engcolileyo okanye i-cryotube yokugcinelwa |
Amanzi | Kwisampulu enesixa esilinganiselweyo se-microbial, njengamanzi epompo, amanzi omthombo, njl., Qokelela ubuncinci be-1 L yamanzi kwaye udlule kwi-0.22 μm yokucoca ukutyebisa i-microbial kwi-membrane.Gcina inwebu kwityhubhu enyumba. |
Ulusu | Khulula ngononophelo umphezulu wolusu ngeswab yomqhaphu engcolileyo okanye ngencakuba yotyando kwaye uyibeke kwityhubhu engenanto. |
Umkhenkce iisampulu kulwelo nitrogen iiyure 3-4 kwaye ugcine kulwelo nitrogen okanye -80 degree ukuya ugcino ixesha elide.Isampulu yokuthumela ngenqanawe eyomileyo iyafuneka.
1.I-Histogram: Ukuhanjiswa kweentlobo
2.Ufuzo olusebenzayo oluchazwe kwiindlela ze-metabolic ze-KEGG
3.Imephu yobushushu: Imisebenzi eyohlukileyo esekelwe kubuninzi bemfuzaI-4.I-Circos ye-CARD i-antibiotic resistance genenes
Ityala le-BMK
Ukuxhaphaka kofuzo lokuxhathisa amayeza okubulala iintsholongwane kunye neentsholongwane zebhaktiriya ecaleni kwengcambu yommango
Ipapashiwe:Ijenali yeZixhobo ezinobungozi, ngo-2021
Iqhinga lokulandelelanisa:
Izinto: Izicatshulwa ze-DNA zamaqhekeza amane eengcambu zemangrove ezinxulumene neesampuli: umhlaba ongatyalwanga, irhizosphere, i-episphere kunye ne-endosphere compartments
Iqonga: Illumina HiSeq 2500
Ujoliso: iMetagenome
16S rRNA gene V3-V4 ummandla
Iziphumo eziphambili
Ulandelelwano lwe-Metagenomic kunye ne-metabarcoding profiling kwi-root-root continuum ye-mangrove saplings yacutshungulwa ukuze kufundwe ukusasazwa kofuzo lwe-antibiotic resistance (ARGs) ukusuka emhlabeni ukuya kwizityalo.Idatha ye-Metagenomic ibonise ukuba i-91.4% yejene yokumelana namayeza okubulala iintsholongwane aye achongwa ngokuqhelekileyo kuwo onke amacandelo omhlaba akhankanywe ngasentla, abonisa ifashoni eqhubekayo.Ulandelelwano lwe-amplicon ye-16S rRNA yenze i-29,285 yokulandelelana, emele iintlobo ze-346.Ukudibanisa neentlobo zeprofayili ngokulandelelana kwe-amplicon, oku kusasazwa kwafunyaniswa kuzimeleyo kwi-microbiota ehambelana neengcambu, nangona kunjalo, inokuququzelelwa yi-mobile ye-genetic elements.Olu phononongo luchonge ukuhamba kwee-ARG kunye neentsholongwane ezibangela izifo ezisuka emhlabeni zisiya kwizityalo ngokudityaniswa kweengcambu zomhlaba.
Isalathiso
Wang, C. , Hu, R. , Strong, PJ , Zhuang, W. , & Shu, L. .(2020).Ukuxhaphaka kwejini yokuxhathisa amayeza okubulala iintsholongwane kunye neentsholongwane ezibangela iintsholongwane ecaleni kwengcambu yommango.Ijenali yeZixhobo ezinobungozi, 408, 124985.