Bulked Segregant kusanthula
Ubwino wa Utumiki
Takagi et al., The plant magazine, 2013
● Kukhazikika kolondola: Kusakaniza zochulukira ndi anthu 30+30 mpaka 200+200 kuti muchepetse phokoso lakumbuyo;zolosera za dera losafanana ndi mawu a mutatants.
● Kusanthula kwathunthu: Kufotokozera mozama kwa jini, kuphatikizapo NR, SwissProt, GO, KEGG, COG, KOG, etc.
● Nthawi yosinthira mwachangu: Kusintha kwa majini mwachangu mkati mwa masiku 45 ogwira ntchito.
● Zokumana nazo zambiri: BMK yathandiza kwambiri m'makhalidwe masauzande ambiri, kuphatikizapo zamoyo zosiyanasiyana monga mbewu, zam'madzi, nkhalango, maluwa, zipatso, ndi zina zotero.
Mafotokozedwe a Utumiki
Chiwerengero cha anthu:
Kugawa mbadwa za makolo omwe ali ndi ma phenotypes otsutsana.
mwachitsanzo F2, Backcrossing (BC), Recombinant inbred line(RIL)
Kusakaniza dziwe
Kwa mikhalidwe yabwino: 30 mpaka 50 anthu (osachepera 20) / zambiri
Kwa ma tratis ochulukirachulukira: apamwamba 5% mpaka 10% mwa anthu omwe ali ndi phenotypes pagulu lonse (osachepera 30+30).
Kuzama kotsatizana kovomerezeka
Osachepera 20X/kholo ndi 1X/mwana payekha (mwachitsanzo kwa ana osakaniza dziwe la 30+30 payekha, kuya kwake kudzakhala 30X pa chochuluka)
Bioinformatics kusanthula
● Kusinthasintha kwamtundu wonse
● Kukonza deta
● Kuyimba kwa SNP/Indel
● Kuwunikidwa kwa chigawo cha ofuna
● Kufotokozera kwa jini ya ofuna
Zitsanzo Zofunika ndi Kutumiza
Zitsanzo Zofunika:
Nucleotides:
| gDNA chitsanzo | Chitsanzo cha minofu |
| Kuyikirapo: ≥30 ng/μl | Zomera: 1-2 g |
| Kuchuluka: ≥2 μg (Volumu ≥15 μl) | Zinyama: 0.5-1 g |
| Chiyero: OD260/280= 1.6-2.5 | Magazi athunthu: 1.5 ml |
Kuyenda kwa Ntchito Yantchito
Kuyesera kupanga
Kupereka zitsanzo
Kusintha kwa RNA
Kumanga laibulale
Kutsata
Kusanthula deta
Pambuyo pogulitsa ntchito
1.Association analysis based on Euclidean Distance (ED) kuti adziwe chigawo cha ofuna kusankha.Mu chithunzi chotsatira
X-axis: Nambala ya chromosome;Dontho lililonse likuyimira mtengo wa ED wa SNP.Mzere Wakuda umagwirizana ndi mtengo wa ED wokwanira.Mtengo wapamwamba wa ED umasonyeza kugwirizana kwakukulu pakati pa malo ndi phenotype.Mzere wofiyira wofiyira umayimira malire a kulumikizana kwakukulu.
2.Association kusanthula kopanda SNP-index
X-axis: Nambala ya chromosome;Dontho lililonse likuyimira SNP-index value.Mzere wakuda umayimira mtengo wa SNP-index wokwanira.Mtengowo ukakhala waukulu, mgwirizanowo umakhala wofunikira kwambiri.
BMK Case
Mphamvu yayikulu yochulukirapo ya locus Fnl7.1 imayika mapuloteni ochedwa embryogenesis okhudzana ndi kutalika kwa khosi la zipatso mu nkhaka.
Lofalitsidwa: Plant Biotechnology Journal, 2020
Njira yotsatirira:
Makolo (Jin5-508, YN): Genome yonse yotsatizana ndi 34× ndi 20×.
Maiwe a DNA (50 a khosi lalitali ndi 50 a khosi lalifupi): Kutengera 61 × ndi 52 ×
Zotsatira zazikulu
Mu phunziro ili, kulekanitsa chiwerengero cha anthu (F2 ndi F2: 3) chinapangidwa ndi kuwoloka khosi lalitali nkhaka mzere Jin5-508 ndi lalifupi khosi YN.Maiwe awiri a DNA adapangidwa ndi anthu 50 akhosi lalitali kwambiri komanso anthu 50 okhala ndi khosi lalifupi kwambiri.Zotsatira zazikulu za QTL zidadziwika pa Chr07 ndi kusanthula kwa BSA ndi mapu achikhalidwe a QTL.Dera la ofuna kusankhidwa linachepetsedwanso ndi kupanga mapu abwino, kuchuluka kwa mawu a jini ndi kuyesa kwa transgenic, komwe kunavumbulutsa jini yofunikira pakuwongolera kutalika kwa khosi, CsFnl7.1.Kuphatikiza apo, polymorphism m'chigawo cholimbikitsa cha CsFnl7.1 idapezeka kuti ikugwirizana ndi mawu ofanana.Kusanthula kwina kwa phylogenetic kunawonetsa kuti Fnl7.1 locus ikuyenera kuti idachokera ku India.
 Mapu a QTL pakuwunika kwa BSA kuti azindikire madera omwe akugwirizana ndi kutalika kwa khosi la nkhaka |  Mbiri za LOD za QTL kutalika kwa nkhaka pakhosi zodziwika pa Chr07 |
Xu, X., ndi al."Chotsatira chachikulu cha kuchuluka kwa Fnl7.1 chimayika mapuloteni ochedwa embryogenesis okhudzana ndi kutalika kwa khosi la zipatso mu nkhaka."Plant Biotechnology Journal 18.7 (2020).
