Kayan tsirrai da ƙwayoyin cuta
Dr. Pat Brown ne ya samar da taswirar yawan mutanen dawaki da aka sani da yawan masu canza sorghum (SCP) a Jami'ar Illinois (wanda yanzu yake a UC Davis). An riga an bayyana shi kuma tarin layuka ne daban-daban da aka canza zuwa rashin jin daɗin lokacin daukar hoto da ƙaramin tsayi don sauƙaƙe girma da haɓaka shuke-shuke a cikin muhallin Amurka. An yi amfani da layuka 510 daga wannan yawan a cikin wannan binciken kodayake saboda mummunan germination da sauran matsalolin kula da inganci, ba duk layukan aka yi amfani da su ba wajen nazarin dukkan halaye uku. A ƙarshe an yi amfani da bayanai daga layuka 345 don nazarin martanin chitin, layuka 472 don amsawar flg22, da 456 don juriyar TLS.B. cookeiAn samo nau'in LSLP18 daga Dr. Burt Bluhm a Jami'ar Arkansas.
Ma'aunin amsawar MAMP
An yi amfani da nau'ikan MAMP guda biyu daban-daban a cikin wannan binciken flg22, (Genscript catalog# RP19986), da chitin. An shuka shuke-shuken dawa a cikin kayan da aka sanya a kan filaye da aka cika da ƙasa (33% Sunshine Redi-Earth Pro Growing Mix) a cikin gidan kore. An shayar da shuke-shuken a ranar da ta gabaci tattara samfurin don guje wa ƙarin danshi a ranar da aka tattara.
An yi bazuwar layukan kuma, saboda dalilai na dabaru, an dasa su a cikin rukuni-rukuni na layuka 60. Ga kowane layi, an dasa 'tukwane' uku da iri biyu a kowane layi. An dasa rukuni-rukuni na gaba da zarar an sarrafa rukunin da ya gabata har sai an tantance dukkan jama'a. An gudanar da gwaje-gwaje guda biyu don duka MAMPs tare da sake yin bazuwar genotypes a cikin kowane zagaye biyu.
An gudanar da gwajin ROS kamar yadda aka bayyana a baya. A takaice, ga kowane layi, an shuka iri shida a cikin tukwane 3 daban-daban. Daga cikin tsinken da aka samu, an zaɓi uku bisa ga daidaito. Ba a yi amfani da tsinken da ya yi kama da ba a saba gani ba ko kuma ya fi tsayi ko gajere fiye da yawancin ba. An cire faifan ganye guda huɗu masu diamita 3 mm daga mafi faɗin ganyen 4 na shuke-shuken dawa guda uku daban-daban na kwanaki 15. Faifan ɗaya a kowane ganye daga shuke-shuke biyu da faifan biyu daga shuka ɗaya, tare da faifan na biyu ya zama mai sarrafa ruwa (duba ƙasa). An yi shawagi da faifan daban-daban a kan 50 µl H20 a cikin faranti mai bakin rijiya 96, an rufe shi da hatimin aluminum don guje wa fallasa ga haske, kuma an ajiye su a zafin ɗaki na dare ɗaya. Washegari da safe an yi maganin amsawa ta amfani da 2 mg/ml chemiluminescent probe L-012 (Wako, kundin adireshi # 120-04891), 2 mg/ml horseradish peroxidase (Nau'in VI-A, Sigma-Aldrich, kundin adireshi # P6782), da 100 mg/ml Chitin ko 2 μM na Flg22. An ƙara 50 µl na wannan maganin amsawa a cikin rijiyoyi uku daga cikin huɗu. Rijiyar ta huɗu ita ce maganin sarrafawa, wanda aka ƙara maganin amsawar ban da MAMP. An haɗa rijiyoyi huɗu marasa komai waɗanda ke ɗauke da ruwa kawai a cikin kowane faranti.
Bayan ƙara maganin amsawa, an auna hasken ta amfani da na'urar karanta microplate mai gano abubuwa da yawa ta SynergyTM 2 (BioTek) kowace minti 2 na tsawon awa 1. Mai karanta farantin yana ɗaukar ma'aunin haske a kowace minti 2 a cikin wannan awa 1. An ƙididdige jimillar duk karatun 31 don ba da ƙimar kowace rijiya. An ƙididdige ƙimar da aka kiyasta don amsawar MAMP ga kowane nau'in genotype kamar (matsakaicin ƙimar haske na rijiyoyin gwaji guda uku - ƙimar rijiyar gwaji) - ban da matsakaicin ƙimar rijiyar mara komai. Ƙimar rijiyar mara komai tana kusa da sifili.
Faifan ganye naNicotiana bentamiana, layin dawaki mai amsawa mai ƙarfi ɗaya (SC0003), da layin dawaki mai amsawa mai ƙarancin amsawa ɗaya (PI 6069) an kuma haɗa su a matsayin masu sarrafawa a cikin kowace faranti mai rijiyoyi 96 don dalilan kula da inganci.
B. cookeiShirye-shiryen allurar rigakafi da allurar rigakafi
B. cookeiAn shirya inoculum kamar yadda aka bayyana a baya. A takaice, an jiƙa hatsin dawa a cikin ruwa na tsawon kwana uku, an wanke, an zuba a cikin kwalba mai siffar konkoli lita 1 sannan a rufe shi na tsawon awa ɗaya a zafin 15psi da 121 °C. Sannan aka yi wa hatsin allurar kimanin 5 ml na mycelia da aka yanka daga sabon shukarB. cookeiAna cire LSLP18 sannan a bar shi na tsawon makonni 2 a zafin ɗaki, ana girgiza kwalaben bayan kwana 3. Bayan makonni 2, an busar da hatsin dawa da naman gwari da ya mamaye a iska sannan a adana a zafin 4°C har sai an yi allurar a gona. An yi amfani da wannan allurar a duk lokacin gwajin kuma an yi sabo kowace shekara. Don yin allurar, an sanya hatsi 6-10 da suka kamu a cikin ciyawar dawa da suka tsufa na tsawon makonni 4-5. Kwayoyin da aka samar daga waɗannan fungi sun fara kamuwa da cuta a cikin ƙananan shuke-shuken dawa cikin mako guda.
Shirya iri
Kafin a dasa iri a gona, an yi wa iri na dawa magani da maganin kashe kwari, maganin kwari, da kuma cakuda mai aminci wanda ya ƙunshi ~ 1% maganin kashe kwari na Spirato 480 FS, 4% maganin kashe kwari na Sebring 480 FS, 3% maganin kariya daga iri na Sorpro 940 ES. Sannan an busar da iri a iska na tsawon kwana 3 wanda hakan ya ba da siraran shafi na wannan cakuda a kusa da iri. Mafi aminci ya ba da damar amfani da maganin kashe kwari na Dual Magnum a matsayin maganin kafin a fara kamuwa da cutar.
Kimanta juriyar Tabon Ganye na Target
An dasa SCP a Cibiyar Bincike ta Central Crops da ke Clayton, NC a ranakun 14-15 ga Yuni 2017 da 20 ga Yuni, 2018 a cikin tsari mai cikakken tsari na tubali tare da kwafi biyu na gwaji a kowane yanayi. An dasa gwaje-gwajen a layuka guda ɗaya na mita 1.8 tare da faɗin layi na mita 0.9 ta amfani da tsaba 10 a kowane fili. An dasa layuka biyu na iyakoki a kusa da gefen kowace gwaji don hana tasirin gefen. An yi allurar gwaje-gwajen a ranakun 20 ga Yuli, 2017 da 20 ga Yuli, 2018 a lokacin da tsire-tsire na dawa suka kasance a matakin girma na 3. An ɗauki ƙima akan sikelin ɗaya zuwa tara, inda aka sami tsire-tsire da ba su nuna alamun cuta ba yayin da aka sami maki tara kuma matattu gaba ɗaya a matsayin ɗaya. An ɗauki ƙima biyu a cikin 2017 da karatu huɗu a cikin 2018 farawa makonni biyu bayan an yi allurar kowace shekara. An ƙididdige sAUDPC (yanki mai daidaitaccen tsari a ƙarƙashin lanƙwasa ci gaban cuta) kamar yadda aka bayyana a baya.
Lokacin Saƙo: Afrilu-01-2021