Zomera ndi zinthu zoyambitsa matenda
Kujambula mitundu ya mbewu za m'gulu la manyuchi komwe kumadziwika kuti sorghum conversion population (SCP) kunaperekedwa ndi Dr. Pat Brown ku University of Illinois (tsopano ku UC Davis). Kwafotokozedwa kale ndipo ndi gulu la mizere yosiyanasiyana yomwe inasinthidwa kukhala yosakhudzidwa ndi kuwala kwa dzuwa komanso yaying'ono kuti ithandize kukula ndi chitukuko cha zomera m'malo ozungulira US. Mizere 510 kuchokera ku mitundu iyi inagwiritsidwa ntchito mu kafukufukuyu ngakhale chifukwa cha kumera koyipa ndi mavuto ena owongolera khalidwe, si mizere yonse yomwe inagwiritsidwa ntchito pofufuza makhalidwe onse atatu. Pomaliza, deta kuchokera ku mizere 345 idagwiritsidwa ntchito pofufuza yankho la chitin, mizere 472 ya yankho la flg22, ndi 456 ya kukana kwa TLS.B. cookeiMtundu wa LSLP18 unapezeka kwa Dr. Burt Bluhm ku University of Arkansas.
Muyeso wa mayankho a MAMP
MAMP ziwiri zosiyana zinagwiritsidwa ntchito mu kafukufukuyu flg22, (Genscript catalog# RP19986), ndi chitin. Zomera za sorghum zinalimidwa m'malo otetezedwa omwe anaikidwa panthaka yodzaza ndi dothi (33% Sunshine Redi-Earth Pro Growing Mix) mu greenhouse. Zomera zinathiriridwa tsiku lisanafike tsiku lotolera zitsanzo kuti zipewe chinyezi chowonjezera pa tsiku lotolera.
Mizereyo inabzalidwa mwachisawawa ndipo, pazifukwa za kayendedwe ka zinthu, inabzalidwa m'magulu a mizere 60. Pa mzere uliwonse, 'miphika' itatu inabzalidwa ndi mbewu ziwiri pa mzere uliwonse. Mizere yotsatira inabzalidwa mwamsanga pamene gulu lapitalo linakonzedwa mpaka anthu onse atayesedwa. Kuyesa kawiri kunachitika pa mitundu yonse ya MAMP ndipo majini anasinthidwanso mwachisawawa pa miyeso iwiri iliyonse.
Mayeso a ROS anachitika monga momwe tafotokozera kale. Mwachidule, pa mzere uliwonse, mbewu zisanu ndi chimodzi zinabzalidwa m'miphika itatu yosiyanasiyana. Kuchokera ku mbande zomwe zinatuluka, zitatu zinasankhidwa kutengera kufanana. Mbande zomwe zinkaoneka zachilendo kapena zazitali kwambiri kapena zazifupi kuposa zambiri sizinagwiritsidwe ntchito. Ma disc anayi a masamba a 3 mm m'mimba mwake adachotsedwa kuchokera ku gawo lalikulu la tsamba lachinayi la zomera zitatu zosiyanasiyana za masiku 15 za manyuchi. Disc imodzi pa tsamba lililonse kuchokera ku zomera ziwiri ndi ma disc awiri kuchokera ku chomera chimodzi, ndipo disc yachiwiri idakhala chowongolera madzi (onani pansipa). Ma disc onsewa adayandama pa 50 µl H20 mu mbale yakuda ya 96-chitsime, yotsekedwa ndi chisindikizo cha aluminiyamu kuti asawononge kuwala, ndipo adasungidwa kutentha kwa chipinda usiku wonse. Mmawa wotsatira njira yothetsera vutoli inapangidwa pogwiritsa ntchito 2 mg/ml chemiluminescent probe L-012 (Wako, catalog # 120-04891), 2 mg/ml horseradish peroxidase (Mtundu VI-A, Sigma-Aldrich, catalog # P6782), ndi 100 mg/ml Chitin kapena 2 μM ya Flg22. 50 µl ya yankho la vutoli inawonjezedwa ku zitsime zitatu mwa zinayi. Chitsime chachinayi chinali chowongolera, chomwe njira yothetsera vutoli kupatulapo MAMP inawonjezedwa. Zitsime zinayi zopanda kanthu zokhala ndi madzi okha zinaphatikizidwanso mu mbale iliyonse.
Pambuyo powonjezera yankho la reaction, kuwala kunayesedwa pogwiritsa ntchito SynergyTM 2 multi-detection microplate reader (BioTek) mphindi ziwiri zilizonse kwa ola limodzi. Wowerenga mbale amayesa kuwala mphindi ziwiri zilizonse mkati mwa ola limodzi ili. Chiwerengero cha mawerengedwe onse 31 chinawerengedwa kuti chipereke mtengo wa chitsime chilichonse. Mtengo woyerekeza wa yankho la MAMP pa genotype iliyonse unawerengedwa ngati (mtengo wapakati wa kuwala kwa zitsime zitatu zoyesera—mtengo wa mock chitsime) -kupatula mtengo wapakati wa chitsime chopanda kanthu. Mitengo ya chitsime chopanda kanthu inali pafupi ndi zero nthawi zonse.
Ma diski a masambaNicotiana benthamiana, mzere umodzi wa manyuchi woyankha bwino (SC0003), ndi mzere umodzi wa manyuchi woyankha bwino (PI 6069) nawonso adaphatikizidwa ngati zowongolera mu mbale iliyonse ya zitsime 96 kuti azilamulira khalidwe.
B. cookeiKukonzekera ndi kuyika katemera wa inoculum
B. cookeiInoculum inakonzedwa monga tafotokozera kale. Mwachidule, tirigu wa manyuchi anaviikidwa m'madzi kwa masiku atatu, kutsukidwa, kuikidwa mu mabotolo a conical a 1L ndikuikidwa mu autoclave kwa ola limodzi pa kutentha kwa 15psi ndi 121 °C. Kenako tiriguwo anaikidwa mu katemera wa pafupifupi 5 ml wa macelia ochokera ku mbewu zatsopano zaB. cookeiLSLP18 imachotsa mbewuzo ndikuzisiya kwa milungu iwiri kutentha kwa chipinda, ndikugwedeza mabotolo masiku atatu aliwonse. Pambuyo pa milungu iwiri, mbewu za manyuchi zomwe zinali ndi bowa zinkaumitsa mpweya kenako n’kusungidwa pa 4 °C mpaka zitapatsidwa katemera m’munda. Katemera yemweyo anagwiritsidwa ntchito pa nthawi yonse yoyesera ndipo anapangidwa kukhala watsopano chaka chilichonse. Popereka katemera, mbewu 6-10 zomwe zinali ndi bowa zinayikidwa mu whorl ya zomera za manyuchi za masabata 4-5. Tizilombo toyambitsa matenda tomwe timapangidwa kuchokera ku bowa timayambitsa matenda m’mbewu za manyuchi zazing’ono mkati mwa sabata imodzi.
Kukonzekera mbewu
Mbeu za manyuchi zisanabzalidwe m'munda, mbewuzo zinkapatsidwa mankhwala ophera fungicide, mankhwala ophera tizilombo, ndi mankhwala ophera tizilombo okhala ndi ~ 1% Spirato 480 FS fungicide, 4% Sebring 480 FS fungicide, 3% Sorpro 940 ES seed safener. Kenako mbewuzo zinaumitsidwa ndi mpweya kwa masiku atatu zomwe zinapangitsa kuti chisakanizochi chikhale chopyapyala mozungulira mbewuzo. Safener inalola kugwiritsa ntchito mankhwala ophera tizilombo otchedwa Dual Magnum ngati mankhwala asanayambe kumera.
Kuwunika kwa kukana kwa malo a Target Leaf
SCP idabzalidwa ku Central Crops Research Station ku Clayton, NC pa June 14–15 2017 ndi June 20, 2018 mwadongosolo lathunthu lopangidwa mwachisawawa ndi zobwerezabwereza ziwiri zoyesera pazochitika zonse. Zoyesera zidabzalidwa m'mizere umodzi ya 1.8 m ndi mulifupi wa mzere wa 0.9 m pogwiritsa ntchito mbewu 10 pa ploti iliyonse. Mizere iwiri ya m'malire idabzalidwa mozungulira mphepete mwa kuyesera kulikonse kuti apewe zotsatirapo zoyipa. Zoyesera zidabzalidwa pa July 20, 2017 ndi July 20, 2018 pomwe mbewu za manyuchi zinali pa siteji ya kukula 3. Ziwerengero zidatengedwa pa sikelo imodzi mpaka zisanu ndi zinayi, pomwe zomera zomwe sizikuwonetsa zizindikiro za matenda zidabzalidwa ngati mbewu zisanu ndi zinayi komanso zakufa kwathunthu zidabzalidwa ngati imodzi. Ziwerengero ziwiri zidabzalidwa mu 2017 ndipo ziwerengero zinayi mu 2018 zidayamba milungu iwiri pambuyo pa kupaka chaka chilichonse. sAUDPC (dera lokhazikika lomwe lili pansi pa curve yopita patsogolo kwa matenda) idawerengedwa monga momwe tafotokozera kale.
Nthawi yotumizira: Epulo-01-2021