Bulked Segregant analysis
Melemo ea Tšebeletso
Takagi et al., Koranta ea semela, 2013
● Sebaka se nepahetseng: Ho kopanya bongata le batho ba 30+30 ho isa ho 200+200 ho fokotsa lerata le ka morao;boitsebahatso ba sebaka sa mokhethoa ea sa amaneng ka mantsoe a tšoanang.
● Tlhahlobo e pharaletseng: Tlhaloso e tebileng ea mosebetsi oa liphatsa tsa lefutso tsa mokhethoa, ho akarelletsa le NR, SwissProt, GO, KEGG, COG, KOG, joalo-joalo.
● Nako ea ho fetola ka potlako: Ho hlahisa liphatsa tsa lefutso ka potlako matsatsing a 45 a ho sebetsa.
● Phihlelo e pharalletseng: BMK e kentse letsoho litšobotsing tse likete-kete tsa tikoloho, e akarelletsang mefuta e sa tšoaneng e kang lijalo, lihlahisoa tsa metsing, meru, lipalesa, litholoana, joalo-joalo.
Litlhaloso tsa Tšebeletso
Baahi:
Ho arola bana ba batsoali ba nang le li-phenotypes tse hanyetsanang.
mohlala, bana ba F2, Backcrossing (BC), Recombinant inbred line(RIL)
Letamo la ho kopanya
Bakeng sa litšobotsi tsa boleng: 30 ho batho ba 50 (bonyane 20) / bongata
Bakeng sa tratis ea bongata: batho ba kaholimo ho 5% ho isa ho 10% ba nang le phenotypes e feteletseng sechabeng kaofela (bonyane 30+30).
Botebo ba tatellano bo khothalelitsoeng
Bonyane 20X/motsoali le 1X/ngoana ka bomong (mohlala, bakeng sa letamo la ho kopanya bana la batho ba 30+30, tatellano ea botebo e tla ba 30X ka bongata)
Litlhahlobo tsa Bioinformatics
● Phetoho eohle ea liphatsa tsa lefutso
● Ts'ebetso ea data
● Ho letsetsa ka SNP/Indel
● Tlhahlobo ea sebaka sa mokhethoa
● Tlhaloso ea mosebetsi oa liphatsa tsa lefutso tsa mokhethoa
Litlhoko tsa Mohlala le Phano
Litlhoko tsa Mohlala:
Li-nucleotides:
| mohlala oa gDNA | Mohlala oa lisele |
| Ho tsepamisa maikutlo: ≥30 ng/μl | Limela: 1-2 g |
| Palo: ≥2 μg (Bolumo ≥15 μl) | Liphoofolo: 0.5-1 g |
| Bohloeki: OD260/280= 1.6-2.5 | Mali a feletseng: 1.5 ml |
Phallo ea Mosebetsi oa Ts'ebeletso
Moqapi oa liteko
Mohlala oa thomello
Tlhahiso ea RNA
Kaho ea laebrari
Tatelano
Tlhahlobo ea lintlha
Litšebeletso tsa ka mor'a thekiso
1.Association analysis based on Euclidean Distance (ED) ho khetholla sebaka sa mokhethoa.Setšoantšong se latelang
X-axis: Nomoro ea chromosome;Letheba ka leng le emela boleng ba ED ba SNP.Mohala o Motšo o lumellana le boleng ba ED bo kentsoeng.Boleng bo phahameng ba ED bo bontša kamano ea bohlokoa haholo pakeng tsa sebaka le phenotype.Mohala o mofubelu o emela moeli oa kopano ea bohlokoa.
2.Asociation analysis e thehiloeng ho se SNP-index
X-axis: Nomoro ea chromosome;Letheba ka leng le emetse boleng ba index ea SNP.Mola o motšo o emetse boleng ba index ea SNP e kentsoeng.Ha boleng bo le boholo, mokhatlo o bohlokoa le ho feta.
Taba ea BMK
The main-effect quantitative trait locus Fnl7.1 e kenyelletsa protheine e ngata ea embryogenesis e amanang le bolelele ba molala oa litholoana ka har'a likomkomere.
E hatisitsoe: Plant Biotechnology Journal, 2020
Leano la tatellano:
Batsoali (Jin5-508, YN): Genome eohle e latellanang bakeng sa 34× le 20×.
Matamo a DNA (50 a molala o molelele le a 50 a melala e mokhutšoane): Ho latela 61× le 52×
Liphetho tsa bohlokoa
Thutong ena, ho arola batho (F2 le F2: 3) ho hlahisitsoe ka ho tšela line ea likomkomere tse molala o molelele Jin5-508 le molala o mokhuts'oane oa YN.Matamo a mabeli a DNA a hahiloe ke batho ba 50 ba melala e telele haholo le batho ba 50 ba melala e mokhuts'oane haholo.QTL ea phello e kholo e ile ea khetholloa ho Chr07 ka tlhahlobo ea BSA le 'mapa oa setso oa QTL.Sebaka sa mokhethoa se ile sa fokotsoa hape ka ho etsa 'mapa o motle, quantification ea liphatsa tsa lefutso le liteko tsa transgenic, tse ileng tsa senola gene ea bohlokoa ho laola bolelele ba molala, CsFnl7.1.Ho feta moo, ho fumanoe hore polymorphism sebakeng sa motšehetsi oa CsFnl7.1 e amana le polelo e lumellanang.Litlhahlobo tse ling tsa phylogenetic li khothalelitse hore sebaka sa Fnl7.1 ho ka etsahala hore ebe se tsoa India.
 QTL-mapa tlhahlobisong ea BSA ho khetholla sebaka sa mokhethoa se amanang le bolelele ba molala oa likomkomere |  Lintlha tsa LOD tsa likomkomere tse bolelele ba molala QTL tse khethiloeng ho Chr07 |
Xu, X. , le al."Tlhahiso e kholo ea sebopeho sa Fnl7.1 e kenyelletsa protheine e ngata ea morao-rao ea embryogenesis e amanang le bolelele ba molala oa litholoana ka har'a likomkomere."Journal ea Biotechnology ea Semela 18.7(2020).
