Izitshalo kanye ne-pathogen materials
Inhlangano yokwenza imephu yenhlangano yamabele eyaziwa ngokuthi isibalo sabantu abaguqula amabele (SCP) inikezwe uDkt. Pat Brown eNyuvesi yase-Illinois (manje ese-UC Davis).Ichazwe ngaphambilini futhi iyiqoqo lemigqa eyahlukahlukene eguqulelwe ekubeni i-photoperiod-insensitivity kanye nesiqu esincane ukuze kube lula ukukhula nokuthuthuka kwezitshalo ezindaweni zase-US.Kusetshenziswe imigqa engu-510 kulesi sibalo sabantu kulolu cwaningo nakuba ngenxa yokuhluma okubi nezinye izinkinga zokulawulwa kwekhwalithi, akuwona wonke imigqa esetshenzisiwe ekuhlaziyeni zonke izici ezintathu.Ekugcineni idatha evela emigqeni engu-345 yasetshenziselwa ukuhlaziywa kwempendulo ye-chitin, imigqa engu-472 yempendulo ye-flg22, kanye ne-456 yokumelana ne-TLS.B. ukhukhiuhlobo lwe-LSLP18 lwatholwa kuDkt. Burt Bluhm eNyuvesi yase-Arkansas.
Isilinganiso sempendulo ye-MAMP
Kusetshenziswe ama-MAMP amabili ahlukene kulolu cwaningo lwe-flg22, (ikhathalogi ye-Genscript# RP19986), kanye ne-chitin .Izitshalo zamabele zazitshalwa zifakwe ezindlini ezigcwele inhlabathi (33% Sunshine Redi-Earth Pro Growing Mix) ku-greenhouse.Izitshalo zaniselwa ngosuku olwandulela ukuqoqwa kwesampula ukugwema umswakama owengeziwe wamaqabunga ngosuku lokuqoqwa.
Imigqa yayingahleliwe futhi, ngenxa yezizathu zokuhlela, yatshalwa ngamaqoqo emigqa engama-60.Emgqeni ngamunye, 'izimbiza' ezintathu zazitshalwe ezinembewu ezimbili emgqeni ngamunye.Amaqoqwana alandelayo atshalwa ngokushesha nje lapho iqoqo langaphambili selicutshunguliwe kwaze kwaba yilapho isihloliwe sonke.Imigijimo emibili yokuhlola yenziwe kuwo womabili ama-MAMP ane-genotypes aphinde ahlelwa ngokungahleliwe ekugijimeni ngakunye kokubili.
Ukuhlolwa kwe-ROS kwenziwa njengoba kuchaziwe ngaphambili.Kafushane, emgqeni ngamunye, imbewu eyisithupha yayitshalwe ezimbizeni ezi-3 ezihlukene.Kulezi zithombo ezitholakele, ezintathu zakhethwa ngokusekelwe ekufaneni.Izithombo ezibukeka zingajwayelekile noma ezinde kakhulu noma ezimfushane kuneziningi azizange zisetshenziswe.Amadiski amaqabunga amane anobubanzi obuyi-3 mm akhishwe engxenyeni ebanzi kakhulu yeqabunga lesi-4 lezitshalo zamabele ezihlukene zezinsuku eziyi-15.Idiski elilodwa eqabungeni ngalinye elivela ezitshalweni ezimbili kanye namadiski amabili esitshalweni esisodwa, idiski yesibili iba ukulawula kwamanzi (bheka ngezansi).Amadiski ayentantiswa ngawodwana ku-50 µl H20 epuleti elimnyama lemithombo engu-96, elivalwe ngesigxivizo se-aluminium ukuze kugwenywe ukuchayeka ekukhanyeni, futhi agcinwe ekamelweni lokushisa ubusuku bonke.Ngakusasa ekuseni isisombululo sokusabela senziwe kusetshenziswa i-2 mg/ml chemiluminescent probe L-012 (Wako, ikhathalogi # 120-04891), 2 mg/ml horseradish peroxidase (Uhlobo VI-A, Sigma-Aldrich, ikhathalogi # P6782), kanye 100 mg/ml Chitin noma 2 μM we-Flg22.I-50 µl yalesi sixazululo sokusabela yengezwe emithonjeni emithathu kwemine.Umthombo wesine wawuwukulawula okuhlekisayo, lapho isisombululo sokusabela singafaki i-MAMP sengezwe.Kwafakwa nemithombo emine engenalutho equkethe amanzi kuphela epuleti ngalinye.
Ngemva kokwengeza isisombululo sokusabela, i-luminescence ikalwa kusetshenziswa i-Synergy TM 2 multi-detection microplate reader (BioTek) njalo ngemizuzu emi-2 ngehora elingu-1.Isifundi sepuleti sithatha izilinganiso ze-luminescence njalo emizuzwini emi-2 ngalelihora elingu-1.Isamba sakho konke ukufundwa okungu-31 kwabalwa ukunikeza inani lomthombo ngamunye.Inani elilinganiselwe lempendulo ye-MAMP ye-genotype ngayinye libalwe njengokuthi (isilinganiso senani le-luminescence lemithombo emithathu yokuhlola—inani lemithombo mbumbulu) -kukhishwe inani lemithombo elingenalutho eliyisilinganiso.Amanani emithombo engenalutho ayehlale esondele kuziro.
Amadiski amaqabunga weNicotiana benthamiana, ulayini wamabele owodwa osabelayo ophezulu (SC0003), kanye nolayini wamabele osabela kancane (PI 6069) nazo zifakwe njengezilawuli epuletini lemithombo engama-96 ngayinye ngezinjongo zokulawula ikhwalithi.
B. ukhukhiukulungiswa kwe-inoculum kanye nokujova
B. ukhukhiinoculum yalungiswa njengoba kuchaziwe ngaphambili.Kafushane, izinhlamvu zamabele zazicwiliswa emanzini izinsuku ezintathu, zihlanjululwe, zifakwe ku-1L we-conical flask futhi zifakwe ngokuzenzakalelayo ihora ku-15psi no-121 °C.Izinhlamvu zabe zigonywa cishe ngo-5 ml we-macerated mycelia ovela kusiko olusha lweB. ukhukhiI-LSLP18 iyazihlukanisa futhi ishiywe amasonto ama-2 ekamelweni lokushisa, inyakazisa amaflask njalo ezinsukwini ezi-3.Ngemuva kwamaviki ama-2, izinhlamvu zamabele ezihlaselwe yisikhunta zomiswa emoyeni bese zigcinwa ku-4 °C kuze kube yilapho zigonywa ensimini.I-inoculum efanayo yasetshenziswa kuso sonke isilingo futhi yenziwa isha njalo ngonyaka.Ukuze kufakwe umjovo, izinhlamvu ezigcwele ezingu-6-10 zazifakwa esihlahleni sezitshalo zamabele ezinamaviki angu-4-5 ubudala.Izinhlamvu ezikhiqizwa kulezi sikhunta zaqala izifo ezitshalweni zamabele ezincane phakathi nesonto.
Ukulungiswa kwembewu
Ngaphambi kokutshalwa kwembewu yamabele ensimini yayilashwa ngesibulali sikhunta, isibulala-zinambuzane, kanye nengxube ephephile equkethe ~ 1% Spirato 480 FS fungicide, 4% Sebring 480 FS fungicide, 3% Sorpro 940 ES seed safener.Khona-ke imbewu yomiswa emoyeni izinsuku ezi-3 okwanikeza ukumbozwa okuncane kwale ngxube ezungeze imbewu.Isisefo savumela ukusetshenziswa komuthi obulala ukhula i-Dual Magnum njengokwelashwa kwangaphambi kokuvela.
Ukuhlolwa kokumelana ne-Target Leaf Spot
I-SCP yatshalwa e-Central Crops Research Station e-Clayton, NC ngoJuni 14–15 2017 nangoJuni 20, 2018 edizayini yebhulokhi engahleliwe enezimpendulo ezimbili zokuhlola esimweni ngasinye.Izivivinyo zatshalwa ku-1.8 m imigqa eyodwa enobubanzi bemigqa engu-0.9 m kusetshenziswa imbewu eyi-10 ngesiqephu ngasinye.Imigqa yomngcele emibili yatshalwa endaweni ezungezile yokuhlolwa ngakunye ukuvimbela imiphumela esemaphethelweni.Ukuhlolwa kwagonywa ngoJulayi 20, 2017 kanye noJulayi 20, 2018 ngaleso sikhathi izitshalo zamabele zase zisezingeni lokukhula 3. Izilinganiso zithathwe esikalini esisodwa kuya kweziyisishiyagalolunye, lapho izitshalo ezingabonisi zimpawu zesifo zatholwa njengeziyisishiyagalolunye futhi ngokuphelele. izitshalo ezifile zatholwa njengenye .Izilinganiso ezimbili zithathwe ku-2017 kanye nokufundwa okune ku-2018 kuqala amasonto amabili ngemva kokujova unyaka ngamunye.I-sAUDPC (indawo ejwayelekile ngaphansi kwejika lokuqhubeka kwesifo) ibalwa njengoba kuchazwe ngaphambilini.
Isikhathi sokuthumela: Apr-01-2021