Izinto zezityalo kunye nezifo
Imephu yeqela le-sorghum association mapping eyaziwa ngokuba yi-sorghum conversion population (SCP) yanikezelwa nguGqr. Pat Brown kwiYunivesithi yase-Illinois (ngoku ese-UC Davis). Ichazwe ngaphambili kwaye yingqokelela yemigca eyahlukeneyo eguqulwe yaba yi-photoperiod-insensitivity kunye nobukhulu obuncinci ukuze kube lula ukukhula nophuhliso lwezityalo kwiindawo zase-US. Imigca engama-510 evela kweli qela isetyenzisiwe kolu phononongo nangona ngenxa yokuhluma okubi kunye neminye imiba yokulawula umgangatho, ayizizo zonke imigca ezisetyenzisiweyo kuhlalutyo lwazo zonke iimpawu ezintathu. Ekugqibeleni idatha evela kwimigca engama-345 isetyenzisiwe kuhlalutyo lwempendulo ye-chitin, imigca engama-472 yempendulo ye-flg22, kunye ne-456 yokumelana ne-TLS.B. cookeiuhlobo lwe-LSLP18 lufunyenwe kuGqr. Burt Bluhm kwiYunivesithi yaseArkansas.
Umlinganiselo wempendulo ye-MAMP
Kusetyenziswe ii-MAMP ezimbini ezahlukeneyo kolu phononongo flg22, (ikhathalogu yeGenscript# RP19986), kunye ne-chitin. Izityalo zesorghum zikhuliswe kwiindawo ezibekwe kwiindawo ezithe tyaba ezizaliswe ngumhlaba (33% Sunshine Redi-Earth Pro Growing Mix) kwigreenhouse. Izityalo zaniselwa ngosuku olungaphambi kokuqokelelwa kweesampuli ukuze kuthintelwe ukufuma okungaphezulu kwamagqabi ngomhla wokuqokelelwa.
Imigca yayingacwangciswanga kwaye, ngenxa yezizathu zezothutho, yatyalwa kwiibhetshi zemigca engama-60. Kumgca ngamnye, 'iimbiza' ezintathu zatyalwa ngembewu ezimbini kumgca ngamnye. Iibhetshi ezilandelayo zatyalwa kwangoko emva kokuba ibhetshi yangaphambili isetyenzisiwe de lonke inani labantu livavanywe. Kwenziwe uvavanyo oluphindwe kabini kuzo zombini iiMAMP kwaye ii-genotypes zaphinda zahlulwahlulwa kwibhetshi nganye kwezi zimbini.
Uvavanyo lweROS lwenziwe njengoko kuchaziwe ngaphambili. Ngamafutshane, kumgca ngamnye, imbewu ezintandathu zatyalwa kwiimbiza ezintathu ezahlukeneyo. Kwizithombo ezivelileyo, ezintathu zakhethwa ngokusekelwe kukulingana. Izithombo ezazibonakala zingaqhelekanga okanye zazinde kakhulu okanye zimfutshane kunezona zininzi azizange zisetyenziswe. Iidiski ezine zamagqabi ezinobubanzi obuyi-3 mm zasuswa kwindawo ebanzi yegqabi lesi-4 lezityalo ezintathu ezahlukeneyo ze-sorghum ezindala ezineentsuku ezili-15. Idiski enye kwigqabi ngalinye kwizityalo ezimbini kunye neediski ezimbini kwisityalo esinye, apho idiski yesibini yaba yindlela yokulawula amanzi (jonga ngezantsi). Iidiski zadada ngazinye kwi-50 µl H20 kwipleyiti emnyama enamaqula angama-96, zitywinwe ngesitywina se-aluminium ukuthintela ukuvezwa ekukhanyeni, kwaye zigcinwe kubushushu begumbi ubusuku bonke. Ngentsasa elandelayo kwenziwa isisombululo sokuphendula kusetyenziswa i-2 mg/ml chemiluminescent probe L-012 (Wako, catalog # 120-04891), i-2 mg/ml horseradish peroxidase (Uhlobo VI-A, Sigma-Aldrich, catalog # P6782), kunye ne-100 mg/ml Chitin okanye i-2 μM yeFlg22. I-50 µl yesi sisombululo sokuphendula yongezwa kwimithombo emithathu kwemine. Imithombo yesine yayiyi-mock control, apho kongezwa isisombululo sokuphendula ngaphandle kweMAMP. Imithombo emine engenanto equlethe amanzi kuphela nayo yafakwa kwipleyiti nganye.
Emva kokongeza isisombululo sempendulo, ukukhanya kulinganiswe kusetyenziswa i-SynergyTM 2 multi-detection microplate reader (BioTek) rhoqo emva kwemizuzu emi-2 ngeyure e-1. I-plate reader ithatha umlinganiselo wokukhanya rhoqo emva kwemizuzu emi-2 ngeli xesha leyure e-1. Isimbuku sazo zonke izifundo ezingama-31 sibalwe ukunika ixabiso lequla ngalinye. Ixabiso eliqikelelweyo lempendulo ye-MAMP kwi-genotype nganye libalwe njenge (ixabiso eliphakathi lokukhanya kwequla leemithombo ezintathu zovavanyo—ixabiso lequla elingabonakaliyo) - ngaphandle kwexabiso eliphakathi lequla elingenanto. Amaxabiso equla elingenanto ayesondele ku-zero rhoqo.
Iidiski zamagqabiUNicotiana benthamiana, umgca omnye we-sorghum ophendula kakhulu (SC0003), kunye nomgca omnye we-sorghum ophendula kancinci (PI 6069) nazo zibandakanyiwe njengezilawuli kwipleyiti nganye yemithombo engama-96 ngeenjongo zokulawula umgangatho.
B. cookeiUkulungiselela kunye nokugonya i-inoculum
B. cookeiIsixokelelwano se-inoculum silungisiwe njengoko kuchaziwe ngaphambili. Ngamafutshane, iinkozo ze-sorghum zifakwe emanzini iintsuku ezintathu, zahlanjwa, zafakwa kwiiflaski ze-conical eziyi-1L zaza zafakwa kwi-autoclave iyure kwi-15psi kunye ne-121 °C. Emva koko iinkozo zafakwa malunga ne-5 ml ye-macerated mycelia evela kwi-culture entsha ye-B. cookeiI-LSLP18 iyayikhupha ize iyishiye iiveki ezi-2 kubushushu begumbi, ishukumisa iiflaski rhoqo emva kweentsuku ezi-3. Emva kweeveki ezi-2, iinkozo ze-sorghum ezihlaselwe yi-fungus zomiswa emoyeni emva koko zigcinwe kwi-4 °C de zifakwe i-incubator yasendle. I-incubator efanayo yasetyenziswa kulo lonke uvavanyo kwaye yenziwa intsha minyaka le. Ekufakeni i-incubator, iinkozo ezi-6-10 ezihlaselwe yi-incubator zafakwa kwi-whorl yezityalo ze-sorghum ezineeveki ezi-4-5 ubudala. Ii-spores eziveliswe kwezi fungi zaqala ukosuleleka kwizityalo ze-sorghum ezincinci kwiveki enye.
Ukulungiswa kwembewu
Ngaphambi kokutyala emasimini, imbewu yesorghum yayiphathwa ngomxube we-fungicide, i-insecticide, kunye ne-safener equlethe i-~ 1% ye-Spirato 480 FS fungicide, i-4% ye-Sebring 480 FS fungicide, i-3% ye-Sorpro 940 ES seed safener. Emva koko imbewu yomiswa ngomoya kangangeentsuku ezi-3 nto leyo eyanika umxube obhityileyo ojikeleze imbewu. I-safener yavumela ukusetyenziswa kwe-herbicide Dual Magnum njengonyango lwangaphambi kokuvela.
Uvavanyo lokuxhathisa amabala eLeaf Target
I-SCP yatyalwa kwiCentral Crops Research Station eClayton, NC nge-14–15 kaJuni 2017 nange-20 kaJuni 2018 kuyilo olupheleleyo lwebhloko olucwangcisiweyo oluneziphindaphindo ezimbini zovavanyo kwimeko nganye. Uvavanyo lwatyalwa kwimigca enye eyi-1.8 m enobubanzi bomqolo obuyi-0.9 m kusetyenziswa imbewu ezili-10 kwiploti nganye. Imigca emibini yomda yatyalwa ngeenxa zonke zovavanyo ngalunye ukuthintela iziphumo zomphetho. Uvavanyo lwatyalwa nge-20 kaJulayi 2017 nange-20 kaJulayi 2018 apho izityalo ze-sorghum zazikwinqanaba lokukhula lesi-3. Uvavanyo lwathathwa kwisikali esinye ukuya kwisithoba, apho izityalo ezingabonisi zimpawu zesifo zanikwa amanqaku njengezityalo ezilithoba nezifileyo ngokupheleleyo zanikwa amanqaku njengesinye. Uvavanyo olubini lwathathwa ngo-2017 kunye nokufundwa okune ngo-2018 okuqala kwiiveki ezimbini emva kotyando minyaka le. I-sAUDPC (indawo emiselweyo phantsi kwe-curve yokuqhubela phambili kwesifo) yabalwa njengoko kuchaziwe ngaphambili.
Ixesha lokuthumela: Epreli-01-2021