Shuka da kayan pathogen
Dr. Pat Brown ne ya samar da yawan taswirar yawan jama'ar dawa da aka fi sani da yawan canjin sorghum (SCP) a Jami'ar Illinois (yanzu a UC Davis).An yi bayaninsa a baya kuma tarin layi ne daban-daban waɗanda aka canza zuwa rashi-rashin hankali da ƙarami don sauƙaƙe girma da haɓaka tsirrai a cikin yanayin Amurka.An yi amfani da layukan 510 daga wannan yawan a cikin wannan binciken ko da yake saboda mummunan germination da sauran batutuwa masu kula da inganci, ba dukkanin layi ba ne aka yi amfani da su wajen nazarin dukkan halaye uku.Daga ƙarshe an yi amfani da bayanai daga layin 345 don nazarin amsawar chitin, layukan 472 don amsa flg22, da 456 don juriyar TLS.B. kukiAn samo nau'in LSLP18 daga Dr. Burt Bluhm a Jami'ar Arkansas.
Ma'aunin amsa MAMP
Anyi amfani da MAMPs daban-daban guda biyu a cikin wannan binciken flg22, (Katalogin Genscript# RP19986), da chitin .An shuka tsire-tsire na dawa a cikin abubuwan da aka shimfiɗa a kan filaye da aka cika da ƙasa (33% Sunshine Redi-Earth Pro Growing Mix) a cikin greenhouse.An shayar da tsire-tsire a ranar da za a tattara samfurin don kauce wa karin danshi ganye a ranar tattarawa.
Layukan sun kasance bazuwar kuma, saboda dalilai na dabaru, an dasa su a cikin batches na layi 60.Ga kowane layi, an dasa 'tukwane' guda uku tare da tsaba biyu a kowane layi.An dasa rukunin na gaba da zarar an gama sarrafa kashin da ya gabata har sai an tantance dukkan jama'a.An gudanar da gwaje-gwajen gwaji guda biyu don duka MAMPs tare da sake fasalin genotypes a cikin kowane gudu biyun.
An gudanar da gwaje-gwajen ROS kamar yadda aka bayyana a baya.A taƙaice, ga kowane layi, an dasa tsaba shida a cikin tukwane 3 daban-daban.Daga cikin tsire-tsire da aka samu, an zaɓi uku bisa ga daidaito.Ba a yi amfani da 'ya'yan da suka yi kama da sabon abu ba ko kuma sun fi tsayi tsayi ko gajarta fiye da yawancin ba a yi amfani da su ba.An cire fayafai guda huɗu na diamita na mm 3 daga mafi faɗin ɓangaren ganye na 4 na tsire-tsire na dawa daban-daban na kwanaki 15 daban-daban.Fayil ɗaya a kowane ganye daga tsire-tsire biyu da fayafai biyu daga shuka ɗaya, tare da diski na biyu ya zama ikon sarrafa ruwa (duba ƙasa).Fayafai an yi su ne daban-daban akan 50 µl H20 a cikin farantin rijiyar 96 baƙar fata, an rufe su da hatimin aluminium don guje wa fallasa zuwa haske, kuma ana kiyaye su a cikin ɗaki na dare.Washegari an yi maganin dauki ta amfani da 2 mg/ml chemiluminescent probe L-012 (Wako, catalog # 120-04891), 2 mg/ml horseradish peroxidase (Nau'in VI-A, Sigma-Aldrich, catalog # P6782), da kuma 100 mg/ml Chitin ko 2 μM na Flg22.50 µl na wannan maganin an ƙara shi zuwa uku daga cikin rijiyoyin huɗu.Rijiyar ta huɗu ita ce sarrafa izgili, wanda aka ƙara maganin amsa ban da MAMP.Haka kuma an hada rijiyoyin da babu ruwa guda hudu dauke da ruwa kawai a cikin kowace faranti.
Bayan ƙara maganin amsawa, an auna haske ta amfani da SynergyTM 2 mai karanta microplate mai yawan ganowa (BioTek) kowane minti 2 don 1 hr.Mai karanta faranti yana ɗaukar ma'aunin haske kowane minti 2 a cikin wannan sa'a 1.An ƙididdige jimlar duk karatun 31 don ba da ƙimar kowace rijiya.An ƙididdige ƙimar ƙimar amsawar MAMP na kowane genotype a matsayin (matsakaicin ƙimar haske na rijiyoyin gwaji uku-ƙimar rijiyar izgili) - debe madaidaicin ƙimar rijiyar mara kyau.Ƙimar rijiyar da ba komai a koyaushe sun kasance kusa da sifili.
Fayilolin ganye naNicotiana benthamiana, Babban layin dawa mai amsawa guda ɗaya (SC0003), da ƙaramin layin dawa ɗaya (PI 6069) kuma an haɗa su azaman sarrafawa a cikin kowane farantin rijiyar 96 don dalilai masu inganci.
B. kukishirye-shiryen inoculum da inoculation
B. kukian shirya inoculum kamar yadda aka bayyana a baya.A taƙaice, an jiƙa hatsin dawa a cikin ruwa na tsawon kwanaki uku, a wanke, a kwashe su a cikin filaye na conical 1L kuma an kwashe su na awa daya a 15psi da 121 ° C.Bayan haka, an shafe hatsin da kusan 5 ml na macerated mycelia daga sabon al'adaB. kukiLSLP18 ya ware kuma ya bar tsawon makonni 2 a zazzabi na ɗaki, yana girgiza flasks kowane kwana 3.Bayan makonni 2, naman gwari da ya kamu da hatsin dawa an bushe shi ta iska sannan a adana shi a zazzabi na 4 ° C har sai an yi aikin gona.An yi amfani da inoculum iri ɗaya don duka gwajin kuma ana yin sabo kowace shekara.Don yin rigakafi, an sanya hatsi 6-10 masu cike da ƙwayar cuta a cikin tsire-tsire na sorghum na mako 4-5.Kwayoyin da aka samu daga waɗannan fungi sun fara kamuwa da cuta a cikin matasan dawa a cikin mako guda.
Shirye-shiryen iri
Kafin dasa iri dawa a cikin gona an bi da su tare da maganin fungicides, maganin kwari, da cakuda mai aminci wanda ya ƙunshi ~ 1% Spirato 480 FS fungicide, 4% Sebring 480 FS fungicide, 3% Sorpro 940 ES mai kare iri.Sa'an nan kuma an bushe tsaba na iska na tsawon kwanaki 3 wanda ya ba da launi na bakin ciki na wannan cakuda a kusa da tsaba.Mai aminci ya ba da damar yin amfani da Dual Magnum na maganin ciyawa azaman maganin riga-kafi.
Kimanta juriya ta Leaf Leaf
An dasa SCP a Cibiyar Binciken Amfanin amfanin gona ta Tsakiya a Clayton, NC a kan Yuni 14-15 2017 da Yuni 20, 2018 a cikin cikakkiyar ƙirar toshe bazuwar tare da maimaita gwaji guda biyu a kowane hali.An dasa gwaje-gwaje a cikin layuka guda 1.8 m tare da faɗin layin 0.9 m ta amfani da tsaba 10 a kowane wuri.An dasa layuka biyu na kan iyaka a kusa da kewayen kowane gwaji don hana tasirin sakamako.An gudanar da gwajin ne a ranar 20 ga Yuli, 2017 da Yuli 20, 2018 a lokacin da tsire-tsire suka kasance a matakin girma na 3. An dauki kididdigar akan ma'auni daya zuwa tara, inda tsire-tsire da ba su da alamun cututtuka sun kasance tara kuma gaba daya. shuke-shuke da suka mutu an saka su a matsayin daya .An ɗauki ƙima biyu a cikin 2017 da karatu huɗu a cikin 2018 farawa makonni biyu bayan inoculation kowace shekara.An ƙididdige sAUDPC (daidaitaccen yanki a ƙarƙashin yanayin ci gaban cuta) kamar yadda aka bayyana a baya.
Lokacin aikawa: Afrilu-01-2021