Zomera ndi tizilombo toyambitsa matenda
Gulu lopanga mapu a manyuchi otchedwa sorghum conversion population (SCP) linaperekedwa ndi Dr. Pat Brown ku yunivesite ya Illinois (tsopano ku UC Davis).Zafotokozedwa kale ndipo ndi mndandanda wa mizere yosiyana siyana yosinthidwa kukhala photoperiod-insensitivity ndi msinkhu wocheperako kuti athandize kukula ndi chitukuko cha zomera ku US.Mizere 510 yochokera ku chiwerengerochi idagwiritsidwa ntchito mu kafukufukuyu ngakhale chifukwa cha kumera koyipa komanso zovuta zina zowongolera bwino, si mizere yonse yomwe idagwiritsidwa ntchito posanthula mikhalidwe yonse itatu.Pamapeto pake deta yochokera ku mizere ya 345 inagwiritsidwa ntchito pofufuza yankho la chitin, mizere ya 472 ya flg22 yankho, ndi 456 ya kukana kwa TLS.B. kuphikastrain LSLP18 inapezedwa kuchokera kwa Dr. Burt Bluhm ku yunivesite ya Arkansas.
Muyezo wa mayankho a MAMP
Ma MAMP awiri osiyana adagwiritsidwa ntchito mu kafukufukuyu flg22, (Genscript catalog# RP19986), ndi chitin .Zomera za manyuchi zimabzalidwa m'malo osungidwa ndi dothi (33% Sunshine Redi-Earth Pro Growing Mix) mu wowonjezera kutentha.Zomera zidathiriridwa dzulo lisanafike kusonkhanitsa zitsanzo kuti zisawonongeke masamba ochulukirapo patsiku lotolera.
Mizereyo idasinthidwa mwachisawawa ndipo, pazifukwa zogwirira ntchito, idabzalidwa m'magulu a mizere 60.Pa mzere uliwonse, 'miphika' itatu inabzalidwa mbewu ziwiri pamzere uliwonse.Magulu otsatirawa adabzalidwa pomwe gulu lapitalo lidakonzedwa mpaka anthu onse atayesedwa.Mayendedwe awiri oyeserera adachitidwa kwa ma MAMP onse okhala ndi ma genotype osinthidwanso mwachisawawa pamayendedwe awiriwo.
Kuyesa kwa ROS kunachitika monga tafotokozera kale.Mwachidule, pa mzere uliwonse, njere zisanu ndi imodzi zinabzalidwa mumiphika itatu yosiyana.Kuchokera pa mbande zomwe zatuluka, zitatu zinasankhidwa potengera kufanana.Mbande zooneka zachilendo kapena zazitali kwambiri kapena zazifupi kuposa zambiri sizinagwiritsidwe ntchito.Masamba anayi a m'mimba mwake a 3 mm anadulidwa kuchokera ku mbali yotakata kwambiri ya tsamba lachinayi la zomera zitatu zosiyana za masiku 15.Disiki imodzi pa tsamba kuchokera ku zomera ziwiri ndi ma diski awiri kuchokera ku chomera chimodzi, ndi chimbale chachiwiri kukhala chowongolera madzi (onani pansipa).Ma discs adayandamitsidwa payekhapayekha pa 50 µl H20 mu mbale yakuda ya 96-chitsime, yosindikizidwa ndi chisindikizo cha aluminiyamu kuti asatengeke ndi kuwala, ndikusungidwa kutentha kwa usiku wonse.M'mawa wotsatira yankho linapangidwa 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 wa Flg22.50 µl ya yankho ili adawonjezeredwa ku zitsime zitatu mwa zinayi.Chitsime chachinayi chinali chiwongolero chonyozeka, chomwe yankho la mayankho osaphatikizapo MAMP linawonjezedwa.Zitsime zinayi zopanda kanthu zokhala ndi madzi okha zinaphatikizidwanso m'mbale iliyonse.
Pambuyo powonjezerapo yankho, luminescence inayesedwa pogwiritsa ntchito Synergy TM 2 multi-detection microplate reader (BioTek) mphindi iliyonse ya 2 kwa 1 hr.Wowerengera mbale amayesa kuyeza kwa luminescence mphindi 2 zilizonse pa 1 h iyi.Chiwerengero cha zowerengera zonse 31 zinawerengedwa kuti zipereke mtengo wa chitsime chilichonse.Mtengo woyerekeza wa mayankho a MAMP pamtundu uliwonse wamtundu wamtundu uliwonse udawerengedwa ngati (mtengo wapakati wa luminescence wa zitsime zitatu zoyesera-mtengo wamtengo wapatali) -kuchotsa mtengo wapakati wopanda kanthu.Zitsime zopanda kanthu zinali pafupi ndi ziro nthawi zonse.
Masamba a masamba aNicotiana benthamiana, chingwe chimodzi cha manyuchi othamanga kwambiri (SC0003), ndi chingwe chimodzi chochepa chotsatira (PI 6069) chinaphatikizidwanso monga zowongolera mu mbale iliyonse ya 96-chitsime pofuna kuwongolera khalidwe.
B. kuphikakukonzekera kwa inoculum ndi kuthirira
B. kuphikainoculum idakonzedwa monga momwe tafotokozera kale.Mwachidule, njere za manyuchi zidaviikidwa m'madzi kwa masiku atatu, kutsukidwa, kutsanulidwa mu 1L conical flasks ndi autoclaved kwa ola limodzi pa 15psi ndi 121 °C.Mbewuzo zidalowetsedwa ndi pafupifupi 5 ml ya macerated mycelia kuchokera ku chikhalidwe chatsopano chaB. kuphikaLSLP18 imadzipatula ndikusiya kwa milungu iwiri kutentha kwa firiji, kugwedeza ma flasks masiku atatu aliwonse.Pambuyo pa milungu iwiri, mbewu za manyuchi zomwe zidakhudzidwa ndi mafangasi zimawumitsidwa ndi mpweya ndikusungidwa pa 4 °C mpaka mutabaya m'munda.Inoculum yomweyi idagwiritsidwa ntchito pa mayeso onse ndikupangidwa mwatsopano chaka chilichonse.Pothirira, mbewu 6-10 zodzala ndi mbewu zimayikidwa mumsewu wa manyowa azaka 4-5.Tinjere timene timatulutsa kuchokera ku mafangawawo tidayambitsa matenda m'zitsamba zazing'ono patangotha sabata imodzi.
Kukonzekera Mbewu
Musanabzale m'munda mbeu ya manyuchi idathiridwa ndi mankhwala opha bowa, mankhwala ophera tizilombo, ndi osakaniza osakaniza omwe ali ndi ~ 1% Spirato 480 FS fungicide, 4% Sebring 480 FS fungicide, 3% Sorpro 940 ES seed safener.Kenako mbewuzo zidawumitsidwa ndi mpweya kwa masiku atatu zomwe zidapereka zokutira zoonda za kusakaniza uku kuzungulira njerezo.Wotetezayo adalola kugwiritsa ntchito herbicide Dual Magnum ngati chithandizo chisanachitike.
Kuwunika kwa Target Leaf Spot resistance
SCP idabzalidwa ku Central Crops Research Station ku Clayton, NC pa June 14-15 2017 ndi June 20, 2018 mumapangidwe athunthu a block omwe ali ndi zoyeserera ziwiri pazochitika zilizonse.Zoyeserera zidabzalidwa m'mizere imodzi ya 1.8m yokhala ndi mzere wa 0.9 m m'lifupi pogwiritsa ntchito njere khumi pagawo lililonse.Mizere iwiri yamalire idabzalidwa mozungulira m'mphepete mwa kuyesa kulikonse kuti mupewe zotsatira za m'mphepete.Mayeserowa adalowetsedwa pa July 20, 2017 ndi July 20, 2018 panthawi yomwe zomera za manyuchi zinali pa siteji ya kukula 3. Miyezo inatengedwa pa sikelo imodzi mpaka zisanu ndi zinayi , kumene zomera zosonyeza kuti palibe zizindikiro za matenda zinayesedwa ngati zisanu ndi zinayi ndipo kwathunthu. zomera zakufa zinawerengedwa ngati chimodzi .Ziwerengero ziwiri zidatengedwa mu 2017 ndipo zowerengera zinayi mu 2018 kuyambira masabata awiri pambuyo pa inoculation chaka chilichonse.sAUDPC (malo okhazikika pansi pa mayendedwe opitilira matenda) adawerengedwa monga tafotokozera kale.
Nthawi yotumiza: Apr-01-2021