Izinto zezitshalo kanye nezifo
Imephu yenhlangano yama-sorghum eyaziwa ngokuthi i-sorghum conversion population (SCP) yanikezwa nguDkt. Pat Brown eNyuvesi yase-Illinois (manje ese-UC Davis). Ichazwe ngaphambilini futhi iyiqoqo lemigqa ehlukahlukene eguqulwe yaba yi-photoperiod-insensitivity kanye nobukhulu obuncane ukuze kube lula ukukhula nokuthuthukiswa kwezitshalo ezindaweni zase-US. Imigqa engu-510 evela kulolu hlanga yasetshenziswa kulolu cwaningo nakuba ngenxa yokuhluma okubi kanye nezinye izinkinga zokulawula ikhwalithi, akuwona wonke imigqa eyasetshenziswa ekuhlaziyeni zonke izici ezintathu. Ekugcineni idatha evela emigqeni engu-345 yasetshenziswa ekuhlaziyeni impendulo ye-chitin, imigqa engu-472 yempendulo ye-flg22, kanye nengu-456 yokumelana ne-TLS.B. ukuphekauhlobo lwe-LSLP18 lutholakale kuDkt. Burt Bluhm eNyuvesi yase-Arkansas.
Ukulinganiswa kwempendulo ye-MAMP
Kusetshenziswe ama-MAMP amabili ahlukene kulolu cwaningo flg22, (ikhathalogi yeGenscript# RP19986), kanye ne-chitin. Izitshalo ze-sorghum zakhuliswa ezindaweni ezibekwe ezicaba ezigcwele inhlabathi (33% Sunshine Redi-Earth Pro Growing Mix) ku-greenhouse. Izitshalo zaniselwa ngosuku olwandulela ukuqoqwa kwesampula ukuze kugwenywe umswakama owengeziwe wamaqabunga ngosuku lokuqoqwa.
Imigqa yayingahleliwe futhi, ngezizathu zezokuthutha, yatshalwa ngamaqoqo emigqa engama-60. Kumugqa ngamunye, kwatshalwa 'izimbiza' ezintathu ngembewu ezimbili ngomugqa ngamunye. Amaqoqo alandelayo atshalwa ngokushesha nje lapho inqwaba yangaphambilini isicutshunguliwe kwaze kwaba yilapho lonke inani labantu selihlolwe. Kwenziwa ukugijima okubili kokuhlola kokubili ama-MAMP kanye nezinhlobo ze-genotypes eziphinde zahlelwa ngokungahleliwe kumugqa ngamunye wemigqa emibili.
Ukuhlolwa kwe-ROS kwenziwa njengoba kuchaziwe ngaphambilini. Kafushane, emgqeni ngamunye, imbewu eziyisithupha zatshalwa ezitsheni ezintathu ezahlukene. Ezitshalweni ezavela, ezintathu zakhethwa ngokusekelwe ekufaneni. Izithombo ezazibukeka zingavamile noma zazinde kakhulu noma zimfushane kuneningi azizange zisetshenziswe. Amadiski amane amaqabunga anobubanzi obungu-3 mm asuswa engxenyeni ebanzi yeqabunga lesi-4 lezitshalo ezintathu ezahlukene ze-sorghum ezindala zezinsuku ezingu-15. Idiski elilodwa ngeqabunga ezitshalweni ezimbili kanye namadiski amabili esitshalweni esisodwa, kanti idiski yesibili yaba yisilawuli samanzi (bheka ngezansi). Amadiski antantwa ngamanye ku-50 µl H20 epuletini elimnyama elinamanzi angu-96, avalwe ngophawu lwe-aluminium ukuze kugwenywe ukuchayeka ekukhanyeni, futhi agcinwa ekushiseni kwegumbi ubusuku bonke. Ngakusasa ekuseni kwenziwa isisombululo sokuphendula kusetshenziswa i-2 mg/ml chemiluminescent probe L-012 (Wako, catalog # 120-04891), i-2 mg/ml horseradish peroxidase (Uhlobo VI-A, Sigma-Aldrich, catalog # P6782), kanye ne-100 mg/ml Chitin noma i-2 μM ye-Flg22. I-50 µl yalesi sixazululo sokuphendula yengezwe emithonjeni emithathu kwemine. Umthombo wesine wawuyi-mock control, lapho kwanezelwa khona isisombululo sokuphendula ngaphandle kwe-MAMP. Imithombo emine engenalutho equkethe amanzi kuphela nayo yafakwa epuletini ngalinye.
Ngemva kokwengeza ikhambi lokusabela, ukukhanya kwalinganiswa kusetshenziswa i-SynergyTM 2 multi-detection microplate reader (BioTek) njalo ngemizuzu emi-2 ihora eli-1. I-plate reader ithatha izilinganiso zokukhanya njalo ngemizuzu emi-2 phakathi naleli hora eli-1. Isamba sazo zonke izifundo ezingu-31 sabalwa ukuze kunikezwe inani lomthombo ngamunye. Inani elilinganisiwe lempendulo ye-MAMP yohlobo ngalunye lwe-genotype labalwa njenge (inani elilinganiselwe lokukhanya kwemithombo emithathu yokuhlola—inani lomthombo ongabonakali) - ngaphandle kwenani elilinganiselwe lomthombo ongenalutho. Amanani omthombo ongenalutho ayesondele ku-zero njalo.
Amadiski amaqabungaNicotiana benthamiana, umugqa owodwa we-sorghum ophendula kakhulu (SC0003), kanye nomugqa owodwa we-sorghum ophendula kancane (PI 6069) nawo afakiwe njengezilawuli epuletini ngalinye lemithombo engu-96 ngezinjongo zokulawula ikhwalithi.
B. ukuphekaukulungiswa kwe-inoculum kanye nokugoma
B. ukuphekaI-inoculum yalungiswa njengoba kuchaziwe ngaphambilini. Kafushane, izinhlamvu zamabele zacwiliswa emanzini izinsuku ezintathu, zahlanzwa, zafakwa emaflaskini ayi-1L aqinile futhi zafakwa nge-autoclave ihora elilodwa ku-15psi kanye no-121 °C. Izinhlamvu zabe sezifakwa nge-5 ml ye-macelia ehlutshiwe evela ekukhuleni okusha kwe-B. ukuphekaI-LSLP18 iyahlukanisa bese ishiywa amasonto ama-2 ekushiseni kwegumbi, inyakazisa amaflaski njalo ezinsukwini ezi-3. Ngemva kwamasonto ama-2, izinhlamvu zamabele ezihlaselwe yisikhunta zomiswa emoyeni bese zigcinwa ku-4 °C kuze kube yilapho zigonyelwa ensimini. I-inoculum efanayo yasetshenziswa kulo lonke uvivinyo futhi yenziwa ibe yintsha minyaka yonke. Ekugonyelweni, izinhlamvu ezingu-6-10 ezihlaselwe yisikhunta zafakwa ku-whorl yezitshalo zamabele ezineminyaka engama-4-5 ubudala. Izinhlamvu ezikhiqizwa yilezi fungi zaqala ukutheleleka ezitshalweni zamabele ezincane zingakapheli isonto.
Ukulungiswa kwembewu
Ngaphambi kokutshala ensimini imbewu yamabele yayiphathwa nge-fungicide, i-insecticide, kanye nengxube ye-safener equkethe i-~ 1% ye-Spirato 480 FS fungicide, i-4% ye-Sebring 480 FS fungicide, i-3% ye-Sorpro 940 ES seed safener. Ngemuva kwalokho imbewu yomiswa emoyeni izinsuku ezi-3 okwanikeza ungqimba oluncane lwale ngxube nxazonke zembewu. I-safener yavumela ukusetshenziswa kwe-herbicide Dual Magnum njengokwelashwa kwangaphambi kokuvela.
Ukuhlolwa kokumelana namachashaza eqabunga eliqondiwe
I-SCP yatshalwa eCentral Crops Research Station eClayton, NC ngoJuni 14–15 2017 nangoJuni 20, 2018 ngendlela engahleliwe ephelele yamabhulokhi eneziphindaphindo ezimbili zokuhlola esimweni ngasinye. Ukuhlolwa kwatshalwa emigqeni eyodwa engu-1.8 m enobubanzi bomugqa obungu-0.9 m kusetshenziswa imbewu eyi-10 ngesiqephu ngasinye. Imigqa emibili yomngcele yatshalwa eduze komphetho wokuhlolwa ngakunye ukuvimbela imiphumela yomphetho. Ukuhlolwa kwagonyelwa ngoJulayi 20, 2017 nangoJulayi 20, 2018 lapho izitshalo zamabele zase zisesigabeni sokukhula sesi-3. Ukulinganisa kwathathwa esikalini esisodwa kuya kwesishiyagalolunye, lapho izitshalo ezingabonisi zimpawu zesifo zanikezwa amaphuzu njengezitshalo eziyisishiyagalolunye nezifile ngokuphelele zanikezwa amaphuzu njengesitshalo esisodwa. Ukulinganisa okubili kwathathwa ngo-2017 kanye nokufundwa okune ngo-2018 kwaqala amasonto amabili ngemva kokugonyelwa minyaka yonke. I-sAUDPC (indawo ejwayelekile ngaphansi kwejika lokuqhubeka kwesifo) yabalwa njengoba kuchaziwe ngaphambilini.
Isikhathi sokuthunyelwe: Ephreli-01-2021