Chirimwa uye pathogen zvinhu
Bato remhunga rinogadzira mepu yehuwandu inozivikanwa sehuwandu hwekutendeuka kwemhunga (SCP) yakapihwa naDr. Pat Brown paUniversity yeIllinois (ikozvino paUC Davis).Yakatsanangurwa kare uye iunganidzo yemitsara yakasiyana-siyana yakashandurwa kuita photoperiod-insensitivity uye diki chimiro kufambisa kukura nekusimudzira kwezvirimwa munzvimbo dzeUS.Mitsetse mazana mashanu negumi kubva muhuwandu uhu yakashandiswa muchidzidzo ichi kunyangwe nekuda kwekushata kumera uye dzimwe nyaya dzekudzora kunaka, haisi mitsetse yese yakashandiswa mukuwongorora maitiro ese ari matatu.Pakupedzisira data kubva kumitsara ye345 yakashandiswa kuongororwa kwechitin mhinduro, mitsara ye472 yeflg22 mhinduro, uye 456 yeTLS kuramba.B. cookeistrain LSLP18 yakawanikwa kubva kuna Dr. Burt Bluhm paYunivhesiti yeArkansas.
MAMP mhinduro kuyerwa
MaMAMP maviri akasiyana akashandiswa muchidzidzo ichi flg22, (Genscript catalog# RP19986), uye chitin .Miti yemhunga yairimwa mumipanda yakaiswa pamafurati akazadzwa nevhu (33% Sunshine Redi-Earth Pro Growing Mix) mugreenhouse.Zvirimwa zvakadiridzwa zuva rakapfuura sampuli yekuunganidza kudzivirira hunyoro hwemashizha pazuva rekuunganidza.
Mitsetse yacho yaive isina kurongeka uye, nekuda kwezvikonzero zvekugadzirisa, yakasimwa mumabhechi e60 mitsetse.Pamutsara wega wega, 'hari' nhatu dzakasimwa nemhodzi mbiri pamutsetse.Mabhechi akatevera akadyarwa nekukurumidza apo batch yapfuura yakanga yagadziriswa kusvikira huwandu hwevanhu hwaongororwa.Mijaho miviri yekuedza yakaitwa kune ese maMAMP ane genotypes akadzokororwa mune imwe neimwe yekumhanya maviri.
ROS bvunzo dzakaitwa sezvakatsanangurwa kare.Muchidimbu, pamutsetse wega wega, mbeu nhanhatu dzakadyarwa muhari nhatu dzakasiyana.Kubva pamiti inoguma, zvitatu zvakasarudzwa zvichienderana nekufanana.Mbeu dzairatidzika sezvisina kujairika kana kuti dzakareba zvakanyanya kana kuti pfupi pane dzakawanda hadzina kushandiswa.Madhisiki mana emashizha e 3 mm dhayamita akachekwa kubva pahupamhi hweshizha rechina rezviyo zvitatu zvakasiyana-siyana zvemazuva gumi nemashanu.Disiki rimwe pashizha kubva kumiti miviri uye madhisiki maviri kubva kune imwe chirimwa, nedhisiki rechipiri richiva kutonga kwemvura (ona pazasi).Madhisiki acho akayangarara ega pa50 µl H20 mundiro nhema ye96-tsime, yakavharwa nechisimbiso chealuminium kuti isavhenekerwe nechiedza, uye ichichengetwa pakamuri tembiricha husiku.Mangwanani akatevera mhinduro yakaitwa pachishandiswa 2 mg/ml chemiluminescent probe L-012 (Wako, catalogue # 120-04891), 2 mg/ml horseradish peroxidase (Type VI-A, Sigma-Aldrich, catalog # P6782), uye 100 mg/ml Chitin kana 2 μM yeFlg22.50 µl yemhinduro iyi yekupindura yakawedzerwa kumatatu ematsime mana.Tsime rechina raive rekunyomba, kwairi mhinduro yekusasanganisa iyo MAMP yakawedzerwa.Mundiro imwe neimwe maiiswawo matsime mana aingova nemvura chete.
Mushure mekuwedzera mhinduro yekugadzirisa, luminescence yakayerwa uchishandisa Synergy TM 2 yakawanda-detection microplate reader (BioTek) yega yega 2 min ye1 hr.Muverengi weplate anotora zviyero zve luminescence yega yega 2 min panguva iyi 1 h.Hwerengedzo yezviverengero zvese 31 yakaverengerwa kupa ukoshi hwetsime rimwe nerimwe.Iko kukosha kunofungidzirwa kwemhinduro yeMAMP yegenotype yega yega yakaverengerwa se (avhareji luminescence kukosha kwematsime matatu ekuedzwa-iyo mock well value) -kubvisa avhareji isina chinhu tsime kukosha.Matsime emutsime asina chinhu aigara ari pedyo ne zero.
Madhisiki emashizha eNicotiana benthamiana, mutsara wemhunga wepamusoro unoteerera (SC0003), uye imwe yakaderera inoteerera mutsara wemhunga (PI 6069) yakabatanidzwawo sekutonga mundiro yega yega ye96-tsime yezvinangwa zvekutonga kwehutano.
B. cookeiinoculum kugadzirira uye inoculation
B. cookeiinoculum yakagadzirwa sezvakatsanangurwa kare.Nenguva pfupi, zviyo zvemhunga zvakanyikwa mumvura kwemazuva matatu, zvosukwa, zvoiswa mu 1L conical flasks uye autoclaved kweawa pa15psi uye 121 °C.Mbeu dzakabva dzaiswa jekiseni rinenge 5 ml ye macerated mycelia kubva kutsva tsika yeB. cookeiLSLP18 inozvitsaura uye inosiya kwemavhiki maviri pamhepo tembiricha, ichizunza maflasks ega ega matatu.Mushure memavhiki maviri, fungus yakazadza tsanga dzemhunga dzakaomeswa nemhepo uye dzochengetwa pa4 °C kusvika mumunda unoculation.Iyo inoculum yakafanana yakashandiswa pakuedza kwese uye yakaitwa nyowani gore rega rega.Kuti pave nejekiseni, 6-10 tsanga dzakazadzwa dzakaiswa muhove ye4-5 mavhiki emiti yemhunga.Mbeu dzakagadzirwa kubva kufungus idzi dzakatanga hutachiona mumiti midiki yemhunga mukati mevhiki.
Kugadzirira mbeu
Mbeu yemhunga isati yadyarwa yaigadzirwa nemushonga wefungi, mushonga wezvipembenene, uye musanganiswa unodzivirira une ~ 1% Spirato 480 FS fungicide, 4% Sebring 480 FS fungicide, 3% Sorpro 940 ES seed safener.Zvadaro mhodzi dzakaomeswa nemhepo kwemazuva matatu izvo zvakapa chifukidzo chitete chemusanganiswa uyu wakakomberedza mbeu.Muchengetedzi akabvumira kushandiswa kwemishonga yesora Dual Magnum semushonga usati wamera.
Kuongororwa kweTarget Leaf Spot resistance
Iyo SCP yakadyarwa paCentral Crops Research Station muClayton, NC muna Chikumi 14-15 2017 uye Chikumi 20, 2018 mune randomized yakazara block dhizaini ine maviri ekuyedza kudzokorora mune yega yega.Maedzo akadyarwa mumitsara 1.8 m imwe ine 0.9 m mutsara wakafara pachishandiswa mbeu gumi paplot.Mitsara miviri yemuganhu yakasimwa yakapoteredza nharaunda yekuedza kwega kwega kudzivirira mhedzisiro.Maedzo aya akaiswa jekiseni musi wa20 Chikunguru, 2017 naChikunguru 20, 2018 panguva iyo mbesa dzemhunga dzaive padanho rekukura 3. Kuyera kwakatorwa pachikero chimwe kusvika kupfumbamwe, uko zvinomera zvairatidza kuti hazvina zviratidzo zvechirwere zvakapihwa sepfumbamwe uye zvachose. zvirimwa zvakafa zvakanhwa sechinhu chimwe .Zviyero zviviri zvakatorwa muna 2017 uye mana kuverenga muna 2018 kutanga masvondo maviri mushure mekudzivirira gore negore.sAUDPC (yakajairwa nzvimbo pasi pechirwere chekuwedzera curve) yakaverengerwa sezvakatsanangurwa kare.
Nguva yekutumira: Kubvumbi-01-2021