Izityalo kunye nezixhobo ze-pathogen
Umbutho wamazimba wokwenza imephu yabemi okwaziwa ngokuba ngabemi bokuguqulwa kwamazimba (SCP) unikwe nguGqr. Pat Brown kwiYunivesithi yase-Illinois (ngoku ese-UC Davis).Ichazwe ngaphambili kwaye yingqokelela yemigca eyahlukeneyo eguqulelwe kwi-photoperiod-insensitivity kunye nesiqu esincinci ukuququzelela ukukhula kunye nophuhliso lwezityalo kwiindawo zase-US.Imigca ye-510 evela kolu luntu isetyenziswe kolu phando nangona ngenxa yokuhluma okubi kunye neminye imiba yokulawula umgangatho, kungekhona yonke imigca esetyenzisiweyo kuhlalutyo lwazo zontathu iimpawu.Ekugqibeleni idatha esuka kwimigca ye-345 isetyenziselwa uhlalutyo lwempendulo ye-chitin, imigca ye-472 ye-flg22 impendulo, kunye ne-456 ye-TLS ukumelana.B. ukuphekastrain LSLP18 yafunyanwa Dr. Burt Bluhm kwiYunivesithi Arkansas.
Umlinganiselo wempendulo ye-MAMP
Kusetyenziswe iiMAMP ezimbini ezahlukeneyo kolu phononongo flg22, (Genscript khathalogu# RP19986), kunye nechitin.Izityalo zamazimba zazilinywa zifakwe kwiiflethi ezizaliswe ngumhlaba (33% Sunshine Redi-Earth Pro Growing Mix) kwindlu yokukhulisela izityalo.Izityalo zankcenkceshelwa kusuku olungaphambi kokuqokelelwa kwesampulu ukuthintela ukufuma okongezelelekileyo kwamagqabi ngosuku lokuqokelela.
Imigca yayingacwangciswanga kwaye, ngenxa yezizathu zolungiselelo, yatyalwa kwiibhetshi zemigca engama-60.Kumgca ngamnye, 'iimbiza' ezintathu zazityalwe ngeembewu ezimbini kumgca ngamnye.Iibhetshi ezilandelayo zaye zatyalwa msinyane nje emva kokuba ibhetshi yangaphambili isetyenzisiwe de kuhlolwe uluntu lonke.Imitsi emibini yovavanyo lwenziwe kuzo zombini ii-MAMPs nge-genotypes iphinde yahlelwa kwakhona kwimitsi emibini.
Uvavanyo lwe-ROS lwenziwa njengoko kuchaziwe ngaphambili.Ngokufutshane, kumgca ngamnye, imbewu ezintandathu zatyalwa kwiimbiza ezi-3 ezahlukeneyo.Ukususela kwizithole ezifunyenweyo, ezintathu zakhethwa ngokusekelwe kwindlela efanayo.Izithole ezikhangeleka zingaqhelekanga okanye ezinde kakhulu okanye ezimfutshane kunoninzi azizange zisetyenziswe.Iidiski ezine zamagqabi ezinobubanzi obuyi-3 mm zaye zasikwa kwinxalenye ebanzi yegqabi le-4 lezityalo zamazimba ezintathu ezahlukeneyo zeentsuku ezili-15.Idiski enye kwigqabi ngalinye kwizityalo ezibini kunye neediski ezimbini kwisityalo esinye, kunye nediski yesibini ibe lulawulo lwamanzi (jonga ngezantsi).Iidiski zazidada ngokwazo kwi-50 µl H20 kwipleyiti emnyama ye-96-well, itywinwe ngesitywina se-aluminium ukuphepha ukuvezwa ekukhanyeni, kwaye igcinwe kwiqondo lobushushu legumbi ubusuku bonke.Ngentsasa elandelayo isisombululo sokusabela senziwe kusetyenziswa i-2 mg/ml chemiluminescent probe L-012 (Wako, ikhathalogu # 120-04891), 2 mg/ml horseradish peroxidase (Uhlobo VI-A, Sigma-Aldrich, ikhathalogu # P6782), kunye I-100 mg / ml Chitin okanye i-2 μM ye-Flg22.I-50 µl yesi sisombululo sokusabela yongezwa kumaqula amathathu kwamane.Iqula lesine yayilulawulo oluhlekisayo, apho isisombululo sokusabela ngaphandle kwe-MAMP songezwa.Amaqula amane angenanto anamanzi kuphela aye aqukwa kwipleyiti nganye.
Emva kokongeza isisombululo sokuphendula, i-luminescence yalinganiswa ngokusebenzisa i-Synergy TM 2 i-multi-detection microplate reader (BioTek) yonke i-2 min kwi-1 hr.Umfundi wepleyiti uthatha imilinganiselo ye-luminescence rhoqo ngemizuzu emi-2 ngeli yure i-1.Isimbuku sawo onke ama-31 okufundwayo sabalwa ukunika ixabiso lequla ngalinye.Ixabiso eliqikelelweyo lempendulo ye-MAMP ye-genotype nganye ibalwa njenge (ixabiso eliphakathi kwe-luminescence yamaqula amathathu okulinga-ixabiso elihlekisayo elihlekisayo) -thabatha i-avareji engenanto engenanto.Amaxabiso equla angenanto ayesoloko ekufutshane no-zero.
Iidiski zeLeaf zeUNicotiana benthamiana, umgca we-sorghum ophezulu ophendulayo (SC0003), kunye nomgca we-sorghum ophantsi ophendulayo (PI 6069) nazo zifakwe njengolawulo kwipleyiti nganye engama-96 ngeenjongo zokulawula umgangatho.
B. ukuphekaukulungiswa kwe-inoculum kunye ne-inoculation
B. ukuphekainoculum yalungiswa njengoko kuchaziwe ngaphambili.Ngokufutshane, iinkozo zamazimba zifakwe emanzini iintsuku ezintathu, zihlanjululwe, zifakwe kwi-1L ye-conical flasks kwaye i-autoclaved iyure kwi-15psi kunye ne-121 °C.Iinkozo zaye zatofwa malunga ne-5 ml ye-macerated mycelia evela kwinkcubeko entshaB. ukuphekaI-LSLP18 yodwa kwaye ishiye iiveki ezi-2 kwindawo yobushushu begumbi, ishukumisa iiflaski rhoqo ngeentsuku ezi-3.Emva kweeveki ezi-2, umngundo ohlasele iinkozo zamazimba zomiswa emoyeni zize zigcinwe ku-4 °C de kufakwe isitofu emasimini.Kwakusetyenziswa isitofu esifanayo kulo lonke ulingo kwaye senziwa sitsha minyaka le.Ukugonywa, iinkozo ezi-6-10 ezihlaselweyo zafakwa kwi-whorl yezityalo zamazimba ezineveki ezi-4-5 ubudala.Imbewu eziveliswe kolu mngundo ziqalise ukosuleleka kwizityalo zamazimba amancinci kwisithuba seveki.
Ukulungiswa kwembewu
Phambi kokutyala emasimini imbewu yamazimba yanyangwa ngesibulali zinambuzane, isibulali zinambuzane, kunye nomxube okhuselekileyo one ~ 1% Spirato 480 FS fungicide, 4% Sebring 480 FS fungicide, 3% Sorpro 940 ES isikhuseli sembewu.Emva koko imbewu yomiswa emoyeni kangangeentsuku ezi-3 nto leyo enika umxube obhityileyo walo mxube ujikeleze imbewu.Isikhuseli savumela ukusetyenziswa kweyeza lokutshabalalisa ukhula iDual Magnum njengonyango lwaphambi kokuvela.
Uvavanyo lokumelana neTage Leaf Spot
I-SCP ityalwe kwi-Central Crops Research Station e-Clayton, NC ngoJuni 14-15 2017 kunye noJuni 20, 2018 kwi-design block block kunye neempendulo ezimbini zokulinga kwimeko nganye.Iimvavanyo zatyalwa kwi-1.8 m kumqolo omnye kunye nobubanzi obuyi-0.9 m kumqolo kusetyenziswa iimbewu ezili-10 kwiploti nganye.Imiqolo emibini yomda yatyalwa ngeenxa zonke kumfuniselo ngamnye ukuthintela iziphumo zomda.Iimvavanyo zifakwe ngoJulayi 20, 2017 kunye noJulayi 20, 2018 apho izityalo zamazimba zazikwinqanaba lokukhula 3. Ukulinganisa kuthathwe kwisikali esinye ukuya kwesithoba, apho izityalo ezibonisa ukuba akukho zibonakaliso zesifo zifunyenwe njengethoba kwaye ngokupheleleyo. izityalo ezifileyo zafunyanwa njengenye.Amanqaku amabini athathwe kwi-2017 kunye nokufundwa okune kwi-2018 ukuqala iiveki ezimbini emva kokugonywa ngonyaka.I-sAUDPC (indawo esemgangathweni phantsi kwegophe lokuqhubela phambili kwesifo) ibalwe njengoko kuchaziwe ngaphambili.
Ixesha lokuposa: Apr-01-2021