Iimveliso

Uhlalutyo lwe-Bulked Segregant

Uhlalutyo lwe-bulked segregant (BSA) bubuchule obusetyenziselwa ukuchonga ngokukhawuleza iimpawu zofuzo ezinxulumene nephenotype.Ukuhamba komsebenzi okuphambili kwe-BSA kuqulethe ukukhetha amaqela amabini abantu abaneephenotypes ezichasene kakhulu, ukudibanisa iDNA yabo bonke abantu ukwenza isambuku se-DNA, ukuchonga ulandelelwano olwahlukileyo phakathi kwamachibi amabini.Obu buchule busetyenziswe kakhulu ekuchongeni iimpawu zemfuza ezinxulunyaniswa ngamandla nemfuza ekujoliswe kuyo kwiigenome zezityalo/zezilwanyana.

Qhagamshelana nathi

Iinkcukacha zeNkonzo

Iziphumo zeDemo

Into yokuphonononga

Iinkonzo eziluncedo

Takagi et al., Ijenali yezityalo, ngo-2013

● Indawo echanekileyo: Ukuxuba isambuku kunye nabantu abangama-30+30 ukuya kuma-200+200 ukunciphisa ingxolo yangasemva;non-synonymous mutatant-based uqikelelo lwengingqi umgqatswa.

● Uhlalutyo olubanzi: Inkcazo yomsebenzi wemfuza yomviwa onzulu, kuquka i-NR, i-SwissProt, i-GO, i-KEGG, i-COG, i-KOG, njl.

● Ixesha loTshintsho oluKhawulezayo: Ukwenziwa kwemfuza ekhawulezileyo yalapha ngaphakathi kweentsuku zokusebenza ezingama-45.

● Amava abanzi: I-BMK ibe negalelo kumawaka eempawu zokwakhiwa kwendawo, equka iintlobo ngeentlobo zezityalo, iimveliso zasemanzini, amahlathi, iintyatyambo, iziqhamo, njl.

IiNgcaciso zeNkonzo

Abemi:
Ukwahlula inzala yabazali abaneephenotypes ezichasayo.
umz. Inzala yeF2, iBackcrossing (BC), iRecombinant inbred line(RIL)

Iphuli yokuxuba
Ngeempawu ezisemgangathweni: abantu abangama-30 ukuya kwangama-50 (ubuncinane bangama-20)/isininzi
Kwi-quantitative tratis: i-5% ephezulu ukuya kwi-10% yabantu abane-phenotypes egqithisileyo kubo bonke abantu (ubuncinane 30+30).

Ubunzulu bolandelelwano olucetyiswayo
Ubuncinci 20X/umzali kunye no-1X/inzala yomntu (umzekelo kwidama lokuxuba abantwana abangama-30+30, ubunzulu bolandelelwano buya kuba yi-30X ngobuninzi)

Uhlalutyo lwe-Bioinformatics

● Ulandelelwano lwe-genome epheleleyo

● Ukulungiswa kwedatha

● SNP/Indel ukufowuna

● Uvavanyo lommandla womviwa

● Inkcazo yomsebenzi wemfuza yomviwa

IiMfuno zeSampulu kunye nokuhanjiswa

IiMfuno zeSampulu:

IiNucleotides:

isampuli yegDNA	Isampulu yezicubu
Ugxininiso: ≥30 ng/μl	Izityalo: 1-2 g
Isixa: ≥2 μg (Umqulu ≥15 μl)	Izilwanyana: 0.5-1 g
Ubunyulu: OD260/280= 1.6-2.5	Igazi elipheleleyo: 1.5 ml

Ukuhamba komsebenzi weNkonzo

Uyilo lovavanyo

Ukuhanjiswa kwesampuli

Ukukhutshwa kwe-RNA

Ukwakhiwa kwethala leencwadi

Ukulandelelanisa

Uhlalutyo lwedatha

Iinkonzo emva kokuthengisa

Ngaphambili: Ukuthelekisa iGenomics

Okulandelayo: I-Evolutionary Genetics

1.Isiseko sokuhlalutya i-Association kwi-Euclidean Distance (ED) ukuchonga ummandla womgqatswa.Kulo mfanekiso ulandelayo

I-X-axis: Inombolo yeChromosome;Ichaphaza ngalinye limele ixabiso le-ED le-SNP.Umgca omnyama uhambelana nexabiso le-ED elifakiweyo.Ixabiso eliphezulu le-ED libonisa umbutho obaluleke ngakumbi phakathi kwesayithi kunye ne-phenotype.Umgca wedash obomvu umele umda wonxulumano olubalulekileyo.

mRNA-FLNC-ubude bokufunda-usasazo

2.Uhlalutyo lombutho olusekelwe kwi-SNP-index

I-X-axis: Inombolo yeChromosome;Ichaphaza ngalinye limele ixabiso lesalathisi se-SNP.Umgca omnyama umele ixabiso lesalathisi se-SNP esifakiweyo.Ixabiso elikhulu, okukhona kubaluleke ngakumbi umbutho.

mRNA-Gqibezela-ORF-ubude-kusasazo

Ityala le-BMK

Elona futhe liphambili lobungakanani belocus Fnl7.1 ifaka iprotein ephuma kade embryogenesis enxulunyaniswa nobude bentamo yesiqhamo kwi cucumber.

Ipapashiwe: Ijenali yeBiyoteknoloji yeZityalo, 2020

Iqhinga lokulandelelanisa:

Abazali (Jin5-508, YN): I-genome yonke ilandelelanisa i-34 × kunye ne-20 ×.

Amachibi e-DNA (i-50 intamo ende kunye ne-50 enentamo emfutshane): ilandelelanisa i-61 × kunye ne-52 ×

Iziphumo eziphambili

Kule sifundo, ukwahlula abantu (i-F2 kunye ne-F2: 3) yenziwe ngokuwela umgca we-cucumber yentamo ende i-Jin5-508 kunye ne-YN emfutshane.Amachibi amabini e-DNA akhiwa ngabantu abantamo ende kakhulu abangama-50 kunye nabantu abantamo imfutshane abangama-50.Isiphumo esikhulu se-QTL ichongiwe kwi-Chr07 ngohlalutyo lwe-BSA kunye nemephu ye-QTL yemveli.Ummandla womgqatswa uye wacuthwa ngakumbi ngemephu ecokisekileyo, ubungakanani bokuchazwa kofuzo kunye nemifuniselo ye-transgenic, eveze ufuzo oluphambili ekulawuleni ubude bentamo, i-CsFnl7.1.Ukongezelela, i-polymorphism kwingingqi ye-CsFnl7.1 yomgqugquzeli yafunyanwa idibene nentetho ehambelanayo.Uhlalutyo olongezelelekileyo lwe-phylogenetic lucebise ukuba i-Fnl7.1 locus kusenokwenzeka ukuba ivela eIndiya.

I-PB-egcwele-ubude-RNA-Ukulandelelana-imeko-isifundo

I-QTL-mapping kuhlalutyo lwe-BSA ukuchonga ummandla womviwa ohambelana nobude bentamo yekhukhamba

I-PB-ubude obugcweleyo-i-RNA-enye-i-splicing

Iiprofayili ze-LOD zobude bentamo yekhukhamba ye-QTL ichongwe kwi-Chr07

Isalathiso

Xu, X., et al."Eyona nto iphambili yesiphumo sobungakanani bendawo i-Fnl7.1 ifaka iprotein eninzi ye-embryogenesis ehambelana nobude bentamo yesiqhamo kwi-cucumber."Ijenali yeBiyoteknoloji yeZityalo 18.7(2020).