Ka nānā 'ana i ka Segregant Bulked
Pono lawelawe
Takagi et al., The plant journal, 2013
● Ka hoʻokaʻawale pololei ʻana: Ka hui ʻana i nā puʻupuʻu me 30+30 a 200+200 mau kānaka e hōʻemi i ka walaʻau hope;wanana wahi moho ma muli o ka mutatants like ole.
● Ka nānā 'ana piha: Ka hō'ike kiko'ī hana gene moho hohonu, me NR, SwissProt, GO, KEGG, COG, KOG , etc.
● ʻOi aku ka wikiwiki o ka hoʻololi ʻana: ʻO ka hoʻopili ʻana i nā genes wikiwiki i loko o 45 mau lā hana.
● ʻIke nui: Ua hāʻawi ʻo BMK i nā kaukani o nā ʻano localization, e uhi ana i nā ʻano ʻano like ʻole e like me nā mea kanu, nā huahana wai, nā nahele, nā pua, nā huaʻai, etc.
Nā kikoʻī lawelawe
Ka heluna kanaka:
Ka hoʻokaʻawale ʻana i nā hua o nā mākua me nā phenotypes kū'ē.
laʻa F2 progeny, Backcrossing (BC), Recombinant inbred line (RIL)
loko huikau
No nā hiʻohiʻona maikaʻi: 30 a 50 mau kānaka (ka liʻiliʻi 20) / nui
No nā tratis quantitative: ʻoi aku ka 5% a hiki i ka 10% mau kānaka me nā phenotypes koʻikoʻi i ka heluna holoʻokoʻa (ka liʻiliʻi 30+30).
Manaʻo ʻia ke kaʻina hana hohonu
Ma ka liʻiliʻi he 20X/makua a me 1X/hua hoʻokahi (no ka laʻana, no ka hui pū ʻana o nā keiki he 30+30 kanaka, ʻo ka hohonu o ka hoʻonohonoho ʻana he 30X no ka nui)
Nā loiloi bioinformatics
● Ka hoʻihoʻi ʻana o ka genome holoʻokoʻa
● Hoʻoponopono ʻikepili
● Kāhea ʻia ʻo SNP/Indel
● Kānā ʻāina moho
● Ka moho moho hana anoano
Nā Koina Laʻana a me ka Hoʻouna ʻana
Nā Koina Laʻana:
Nucleotides:
| hōʻailona gDNA | Laʻana lole |
| Hoʻopili: ≥30 ng / μl | Nā mea kanu: 1-2 g |
| Ka nui: ≥2 μg (Volumn ≥15 μl) | Nā holoholona: 0.5-1 g |
| Maʻemaʻe: OD260/280= 1.6-2.5 | ʻO ke koko holoʻokoʻa: 1.5 ml |
Holo Hana Hana
Hoʻolālā hoʻokolohua
Hāʻawi laʻana
RNA unuhi
Kūkulu hale waihona puke
Ke kaʻina ʻana
ʻIkepili ʻikepili
Nā lawelawe ma hope o ke kūʻai aku
1. Hoʻokumu ka ʻikepili hui ma Euclidean Distance (ED) no ka ʻike ʻana i ka ʻāpana moho.Ma keia kii
X-axis: Helu Chromosome;Hōʻike kēlā me kēia kiko i kahi waiwai ED o kahi SNP.Pili ka laina ʻEleʻele me ka waiwai ED kūpono.Hōʻike ka waiwai ED kiʻekiʻe i kahi hui nui ma waena o ka pūnaewele a me ka phenotype.Hōʻike ka laina ʻulaʻula i ka paepae o ka hui nui.
2.Association analysis ma muli o ka SNP-index
X-axis: Helu Chromosome;Hōʻike kēlā me kēia kiko i ka waiwai SNP-index.Kū ka laina ʻeleʻele no ka waiwai SNP-index kūpono.ʻO ka nui o ka waiwai, ʻoi aku ka nui o ka hui.
Hihia BMK
Hoʻopili ka locus Fnl7.1 i ka nui o ka embryogenesis e pili ana me ka lōʻihi o ka ʻāʻī hua i loko o ka kukumba.
Paʻi ʻia: Mea Puke Biotechnology Journal, 2020
Hoʻolālā kaʻina hana:
Nā Mākua (Jin5-508, YN): Ke hoʻomau nei ka genome holoʻokoʻa no 34 × a me 20 ×.
Nā loko DNA (50 ʻāʻī lōʻihi a me 50 ʻāʻī pōkole): Hoʻihoʻi ʻia no 61 × a me 52 ×
Nā hopena koʻikoʻi
Ma kēia haʻawina, hoʻokaʻawale ʻia ka heluna kanaka (F2 a me F2:3) ma ke ala ʻana i ka laina kukumba ʻāʻī lōʻihi Jin5-508 a me ka ʻāʻī pōkole YN.Ua kūkulu ʻia ʻelua mau loko DNA e 50 mau kānaka ʻāʻī lōʻihi loa a me 50 mau kānaka ʻāʻī pōkole loa.Ua ʻike ʻia ka QTL hopena nui ma Chr07 e ka loiloi BSA a me ka palapala palapala QTL kuʻuna.Ua hōʻemi hou ʻia ka ʻāina o ka moho e ka palapala ʻāina maikaʻi, ka helu ʻana o nā ʻano gene a me nā hoʻokolohua transgenic, i hōʻike ʻia i ke ʻano kumu i ka hoʻomalu ʻana i ka lōʻihi o ka ʻāʻī, CsFnl7.1.Eia kekahi, ua ʻike ʻia ka polymorphism ma ka ʻāina hoʻolaha CsFnl7.1 e pili ana me ka ʻōlelo pili.Ua manaʻo ʻia ka loiloi phylogenetic ʻo Fnl7.1 locus no India mai.
 QTL-mapping ma BSA analysis no ka ʻike ʻana i ka ʻāina moho e pili ana me ka lōʻihi o ka ʻāʻī kukama |  ʻIke ʻia nā moʻolelo LOD o ka ʻāʻī kukama-lōʻihi QTL ma Chr07 |
Xu, X. , et al."ʻO ka locus Fnl7.1 ka hopena nui e hoʻopili ai i kahi protein embryogenesis nui e pili ana me ka lōʻihi o ka ʻāʻī hua i loko o ka kukumba."Mea Puke Biotechnology 18.7(2020).
