Ibikoresho n'ibiti bitera indwara
Ihuriro ry’amasaka ryerekana abaturage bazwi ku izina ry’abaturage bahindura amasaka (SCP) ryatanzwe na Dr. Pat Brown muri kaminuza ya Illinois (ubu ni UC Davis).Byasobanuwe mbere kandi ni icyegeranyo cyimirongo itandukanye yahinduwe kuri Photoperiod-sensensitivite hamwe nuburebure buto kugirango byorohereze imikurire niterambere ryibimera mubidukikije muri Amerika.Imirongo 510 yavuye muri aba baturage yakoreshejwe muri ubu bushakashatsi nubwo kubera kumera nabi nibindi bibazo byo kugenzura ubuziranenge, ntabwo imirongo yose yakoreshejwe mugusesengura imico uko ari itatu.Ubwanyuma amakuru yavuye kumurongo 345 yakoreshejwe mugusesengura igisubizo cya chitin, imirongo 472 kubisubizo bya flg22, na 456 kubirwanya TLS.B. cookeiamananiza LSLP18 yakuwe na Dr. Burt Bluhm muri kaminuza ya Arkansas.
Igipimo cyo gusubiza MAMP
MAMPs ebyiri zitandukanye zakoreshejwe muri ubu bushakashatsi flg22, (Cataloge ya Genscript # RP19986), na chitin.Ibihingwa by'amasaka byahinzwe mu gushyiramo amagorofa yuzuyemo ubutaka (33% Izuba Rirashe Redi-Earth Pro Growing Mix) muri parike.Ibimera byavomerewe umunsi umwe mbere yo gukusanya icyitegererezo kugirango birinde ububobere bwamababi kumunsi wo gukusanya.
Imirongo yatoranijwe kandi, kubwimpamvu zifatika, zatewe mubice byimirongo 60.Kuri buri murongo, 'inkono' eshatu zatewe n'imbuto ebyiri kumurongo.Ibice byakurikiyeho byatewe mugihe icyiciro kibanziriza gutunganywa kugeza abaturage bose basuzumwe.Gukoresha ibigeragezo bibiri byakorewe MAMPs zombi hamwe na genotypes yongeye gutondekanya muri buri kwiruka.
ROS isuzuma ryakozwe nkuko byasobanuwe mbere.Muri make, kuri buri murongo, imbuto esheshatu zatewe mu nkono 3 zitandukanye.Mu ngemwe zavuyemo, eshatu zatoranijwe zishingiye ku bumwe.Imbuto zasaga nkidasanzwe cyangwa zari ndende cyane cyangwa ngufi kurenza ubwinshi ntabwo zakoreshejwe.Disiki yamababi ane ya mm 3 ya diametre yacukuwe mugice kinini cyibabi rya 4 ryibiti bitatu byiminsi 15 byamasaka.Disiki imwe kuri buri kibabi kiva mubihingwa bibiri na disiki ebyiri ziva mu gihingwa kimwe, hamwe na disiki ya kabiri ihinduka igenzura ry'amazi (reba hano hepfo).Disiki yareremba kugiti cye kuri 50 µl H20 mumasahani yumukara 96-iriba, igashyirwaho kashe ya aluminiyumu kugirango wirinde urumuri, kandi ikabikwa ubushyuhe bwicyumba ijoro ryose.Bukeye bwaho, igisubizo cyakemuwe hifashishijwe 2 mg / ml ya chemiluminescent probe L-012 (Wako, catalog # 120-04891), 2 mg / ml horseradish peroxidase (Ubwoko VI-A, Sigma-Aldrich, catalog # P6782), na 100 mg / ml Chitine cyangwa 2 μM ya Flg22.50 µl yiki gisubizo cyongeweho kuri bitatu muri bine.Iriba rya kane ryari igenzura ry'agashinyaguro, hiyongereyeho igisubizo cya reaction ukuyemo MAMP.Amariba ane yuzuye arimo amazi gusa nayo yashyizwe muri buri sahani.
Nyuma yo kongeramo igisubizo cyibisubizo, luminescence yapimwe hifashishijwe SynergyTM 2 isoma microplate umusomyi (BioTek) buri min 2 mumasaha 1.Umusomyi wibisahani afata ibipimo bya luminescence buri min 2 muri iyi 1 h.Umubare wibisomwa byose uko ari 31 byabazwe kugirango utange agaciro kuri buri riba.Agaciro kagereranijwe kubisubizo bya MAMP kuri buri genotype yabazwe nk (impuzandengo ya luminescence yagaciro kumariba atatu yubushakashatsi-agaciro keza urw'agashinyaguro) -kugereranya agaciro keza neza.Iriba ryuzuye agaciro kari hafi ya zeru.
Disiki yamababi yaNikotiana benthamiana, umurongo umwe w'amasaka yitabirwa cyane (SC0003), n'umurongo umwe w'amasaka utitabira neza (PI 6069) nawo washyizwemo nk'ubugenzuzi muri buri sahani 96-iriba hagamijwe kugenzura ubuziranenge.
B. cookeigutegura inoculum no gukingirwa
B. cookeiinoculum yateguwe nkuko byasobanuwe mbere.Muri make, ibinyampeke byamasaka byinjijwe mumazi muminsi itatu, byogejwe, bihunikwa mumashanyarazi ya 1L hanyuma bigashyirwaho autoclave kumasaha imwe kuri 15psi na 121 ° C.Ibinyampeke noneho byatewe hamwe na miriyoni 5 za mycelia ya macera kuva mumico mishya yaB. cookeiLSLP18 yitaruye kandi igenda ibyumweru 2 mubushyuhe bwicyumba, kunyeganyeza flas buri minsi 3.Nyuma yibyumweru 2, igihumyo cyanduye ibinyampeke byumye hanyuma bikabikwa kuri 4 ° C kugeza igihe byatewe.Inoculum imwe yakoreshejwe mugeragezwa yose kandi ikorwa buri mwaka.Kubitera, intete zanduye 6-10 zashyizwe muburaya bwibiti byamasaka bimaze ibyumweru 4-5.Intanga ngabo ziva muri utwo duhumyo zatangiye kwandura ibihingwa byamasaka bitarenze icyumweru.
Gutegura imbuto
Mbere yo gutera mu murima imbuto y'amasaka yavuwe hakoreshejwe fungiside, udukoko twica udukoko, hamwe nuruvange rwizewe rurimo ~ 1% Spirato 480 FS fungiside, 4% Sebring 480 FS fungiside, 3% Sorpro 940 ES yangiza imbuto.Noneho imbuto zumishijwe muminsi 3 zitanga igicucu cyoroshye kivanze nimbuto.Umutekano wemereye gukoresha imiti yica ibyatsi Dual Magnum nkumuti wibanze.
Isuzumabumenyi ryintego yibibabi birwanya
SCP yatewe kuri sitasiyo y’ubushakashatsi y’ibihingwa hagati ya Clayton, NC ku ya 14-15 Kamena 2017 na 20 Kamena 2018 mu buryo bwuzuye bwo guhitamo hamwe n’ubushakashatsi bubiri bwakorewe muri buri kibazo.Ubushakashatsi bwatewe muri 1.8 m umurongo umwe ufite ubugari bwa 0.9 m ukoresheje imbuto 10 kuri buri kibanza.Imirongo ibiri yumupaka yatewe hafi ya buri igeragezwa kugirango hirindwe ingaruka.Ubushakashatsi bwatewe ku ya 20 Nyakanga 2017 na 20 Nyakanga 2018 ari naho ibihingwa by’amasaka byari bigeze ku mikurire ya 3. Ibipimo byafashwe ku munzani umwe kugeza ku cyenda, aho ibimera byerekana ko nta kimenyetso cy’indwara byatsinzwe ari icyenda kandi byuzuye ibihingwa byapfuye byatsinzwe kimwe.Ibipimo bibiri byafashwe muri 2017 nibisomwa bine muri 2018 bitangira ibyumweru bibiri nyuma yo guterwa buri mwaka.SAUDPC (agace gasanzwe munsi yindwara igenda itera) yabazwe nkuko byasobanuwe mbere.
Igihe cyo kohereza: Apr-01-2021