Ukusatshalaliswa kwemvula kwesizini eSifundazweni saseGuizhou akulingana, kunemvula eningi entwasahlobo nasehlobo, kodwa izithombo ze-rapeseed zisengozini yokucindezeleka kwesomiso ekwindla nasebusika, okuthinta kakhulu isivuno. I-mustard iyisitshalo esikhethekile sembewu kawoyela esikhula kakhulu eSifundazweni saseGuizhou. Sibekezelela isomiso kakhulu futhi singatshalwa ezindaweni zezintaba. Siwumthombo ocebile wezakhi zofuzo ezingamelani nesomiso. Ukutholakala kwezakhi zofuzo ezingamelani nesomiso kubaluleke kakhulu ekuthuthukiseni izinhlobo zama-mustard. kanye nokusungula izinto ezintsha ezinsizeni ze-germplasm. Umndeni we-GRF udlala indima ebalulekile ekukhuleni nasekuthuthukisweni kwezitshalo kanye nokusabela ekucindezelekeni kwesomiso. Njengamanje, izakhi zofuzo ze-GRF zitholakale ku-Arabidopsis 2, ilayisi (Oryza sativa) 12, i-rapeseed 13, ukotini (Gossypium hirsutum) 14, ukolweni (Triticum). aestivum)15, i-pearl millet (Setaria italica)16 kanye ne-Brassica17, kodwa akukho mibiko yezakhi zofuzo ze-GRF ezitholakale kuma-mustard. Kulolu cwaningo, izakhi zofuzo zomndeni we-GRF ze-mustard zitholakale ezingeni lonke le-genome futhi izici zazo zomzimba nezamakhemikhali, ubudlelwano bokuziphendukela kwemvelo, i-homology, i-motifs egciniwe, isakhiwo sezakhi zofuzo, ukuphindaphindwa kwezakhi zofuzo, izakhi ze-cis kanye nesigaba sokutshala (isigaba samaqabunga amane) kwahlaziywa. Amaphethini okubonakaliswa ngaphansi kokucindezeleka kwesomiso ahlaziywa ngokuphelele ukuze kunikezwe isisekelo sesayensi sezifundo ezengeziwe mayelana nomsebenzi ongaba khona wezakhi zofuzo ze-BjGRF ekuphenduleni kwesomiso kanye nokuhlinzeka ngezakhi zofuzo ezifanelekayo zokuzalanisa i-mustard ebekezelela isomiso.
Izakhi zofuzo ze-BjGRF ezingamashumi amathathu nane zitholakale ku-genome ye-Brassica juncea kusetshenziswa usesho olubili lwe-HMMER, lonke oluqukethe izizinda ze-QLQ kanye ne-WRC. Uchungechunge lwe-CDS lwezakhi zofuzo ze-BjGRF ezikhonjiwe luvezwe kuThebula Elingeziwe S1. I-BjGRF01–BjGRF34 iqanjwe ngokususelwa endaweni yazo ku-chromosome. Izakhiwo ze-physicochemical zalo mndeni zibonisa ukuthi ubude be-amino acid buyahlukahluka kakhulu, kusukela ku-261 aa (BjGRF19) kuya ku-905 aa (BjGRF28). Iphuzu le-isoelectric le-BjGRF lisukela ku-6.19 (BjGRF02) kuya ku-9.35 (BjGRF03) ngesilinganiso esingu-8.33, kanti u-88.24% we-BjGRF uyiprotheni eyisisekelo. Ububanzi besisindo sama-molecule obubikezelwe be-BjGRF busukela ku-29.82 kDa (BjGRF19) kuya ku-102.90 kDa (BjGRF28); inkomba yokungazinzi kwamaprotheni e-BjGRF isukela ku-51.13 (BjGRF08) kuya ku-78.24 (BjGRF19), wonke angaphezulu kuka-40, okubonisa ukuthi inkomba ye-fatty acid isukela ku-43.65 (BjGRF01) kuya ku-78.78 (BjGRF22), isilinganiso se-hydrophilicity (GRAVY) sisukela ku--1.07 (BjGRF31) kuya ku--0.45 (BjGRF22), wonke amaprotheni e-BjGRF aphilicious ane-GRAVY engalungile, okungenzeka ukuthi kungenxa yokuntuleka kwe-hydrophobicity okubangelwa yizinsalela. Ukubikezela indawo engaphansi kweseli kubonise ukuthi amaprotheni angu-31 e-BjGRF afakwe ikhodi angatholakala ku-nucleus, i-BjGRF04 ingatholakala kuma-peroxisome, i-BjGRF25 ingatholakala ku-cytoplasm, kanye ne-BjGRF28 ingatholakala kuma-chloroplast (Ithebula 1), okubonisa ukuthi ama-BjGRF angatholakala ku-nucleus futhi adlale indima ebalulekile yokulawula njengento yokubhala.
Ukuhlaziywa kwe-phylogenetic kwemindeni ye-GRF ezinhlotsheni ezahlukene kungasiza ekufundeni imisebenzi yezakhi zofuzo. Ngakho-ke, ukulandelana kwe-amino acid okugcwele kwama-rapeseed angu-35, i-turnip engu-16, ilayisi engu-12, i-millet engu-10 kanye nama-GRF angu-9 e-Arabidopsis kwalandwa futhi kwakhiwa umuthi we-phylogenetic ngokusekelwe kuma-gene angu-34 e-BjGRF ahlonziwe (Isithombe 1). Imindeni emithathu engaphansi iqukethe inani elihlukile lamalungu; ama-TF angu-116 e-GRF ahlukaniswe ngemindeni emithathu engaphansi (amaqembu A~C), aqukethe ama-59 (50.86%), ama-34 (29.31%) kanye nama-23 (19.83)% ama-GRF, ngokulandelana. Phakathi kwawo, amalungu omndeni we-BjGRF angu-34 ahlakazekile emindenini emi-3 engaphansi: amalungu angu-13 eqenjini A (38.24%), amalungu angu-12 eqenjini B (35.29%) kanye namalungu angu-9 eqenjini C (26.47%). Enkambisweni ye-polyploidization ye-mustard, inani lama-gene e-BjGRF emindenini ehlukene lihlukile, futhi kungenzeka ukuthi ukwandiswa nokulahleka kwezakhi zofuzo kwenzeke. Kubalulekile ukuqaphela ukuthi akukho ukusatshalaliswa kwama-GRF elayisi ne-millet eqenjini C, kuyilapho kukhona ama-GRF amabili elayisi kanye ne-GRF eyodwa ye-millet eqenjini B, futhi iningi lama-GRF elayisi ne-millet ahlanganiswe egatsheni elilodwa, okubonisa ukuthi ama-BjGRF ahlobene kakhulu nama-dicot. Phakathi kwawo, izifundo ezijulile kakhulu ngomsebenzi we-GRF ku-Arabidopsis thaliana zinikeza isisekelo sezifundo ezisebenzayo zama-BjGRF.
Umuthi we-mustard ohlanganisa i-Brassica napus, i-Brassica napus, ilayisi, i-millet kanye namalungu omndeni we-Arabidopsis thaliana GRF.
Ukuhlaziywa kwezakhi zofuzo eziphindaphindayo emndenini we-mustard GRF. Umugqa ompunga ngemuva umele ibhulokhi evumelanisiwe ku-genome ye-mustard, umugqa obomvu umele ukuphindaphinda okuhlukaniswe kabili kwezakhi zofuzo ze-BjGRF;
Ukuvezwa kwezakhi zofuzo ze-BjGRF ngaphansi kokucindezeleka kwesomiso esigabeni sesine samaqabunga. Idatha ye-qRT-PCR iboniswe kuThebula Elingeziwe S5. Umehluko omkhulu kudatha uboniswa ngezinhlamvu ezincane.
Njengoba isimo sezulu somhlaba wonke siqhubeka nokushintsha, ukufunda ukuthi izitshalo zibhekana kanjani nokucindezeleka kwesomiso nokuthuthukisa izindlela zazo zokubekezelelana sekuyinto ebaluleke kakhulu ocwaningweni18. Ngemva kwesomiso, isakhiwo se-morphological, ukubonakaliswa kwezakhi zofuzo kanye nezinqubo ze-metabolic zezitshalo kuzoshintsha, okungaholela ekuphelisweni kwe-photosynthesis nokuphazamiseka kwe-metabolic, okuthinta isivuno kanye nekhwalithi yezitshalo19,20,21. Lapho izitshalo zizwa izimpawu zesomiso, zikhiqiza izithunywa zesibili njenge-Ca2+ kanye ne-phosphatidylinositol, zandise ukuhlushwa kwe-calcium ion yangaphakathi kweseli futhi zisebenze inethiwekhi yokulawula yendlela ye-protein phosphorylation22,23. Iphrotheni yokugcina eqondiwe ihileleke ngqo ekuvikeleni kwamaseli noma ilawula ukubonakaliswa kwezakhi zofuzo zokucindezeleka ezihlobene ngama-TF, okuthuthukisa ukubekezelelana kwezitshalo nokucindezeleka24,25. Ngakho-ke, ama-TF adlala indima ebalulekile ekuphenduleni ukucindezeleka kwesomiso. Ngokusho kokulandelana kanye nezakhiwo zokubopha i-DNA zama-TF asabela ekucindezelweni kwesomiso, ama-TF angahlukaniswa ngemindeni ehlukene, njenge-GRF, ERF, MYB, WRKY kanye neminye imindeni26.
Umndeni wezakhi zofuzo ze-GRF uhlobo lwe-TF ethile yesitshalo edlala indima ebalulekile ezicini ezahlukene njengokukhula, intuthuko, ukudluliselwa kwesignali kanye nezimpendulo zokuzivikela kwezitshalo27. Kusukela kwatholakala i-gene yokuqala ye-GRF ku-O. sativa28, kuye kwatholakala izakhi zofuzo ze-GRF eziningi ezinhlotsheni eziningi futhi kuboniswe ukuthi zithinta ukukhula kwezitshalo, intuthuko kanye nempendulo yokucindezeleka8, 29, 30,31,32. Ngokushicilelwa kochungechunge lwezakhi zofuzo ze-Brassica juncea, ukuhlonza umndeni wezakhi zofuzo ze-BjGRF kwaba nokwenzeka33. Kulolu cwaningo, kwatholakala izakhi zofuzo ze-BjGRF ezingu-34 kuyo yonke i-genome yesinaphi futhi zaqanjwa ngokuthi i-BjGRF01–BjGRF34 ngokusekelwe endaweni yazo ye-chromosome. Zonke ziqukethe izizinda ze-QLQ kanye ne-WRC ezigcinwe kahle kakhulu. Ukuhlaziywa kwezakhiwo ze-physicochemical kubonise ukuthi umehluko ezinombolweni ze-amino acid kanye nesisindo sama-molecule samaprotheni e-BjGRF (ngaphandle kwe-BjGRF28) wawungabalulekile, okubonisa ukuthi amalungu omndeni we-BjGRF angaba nemisebenzi efanayo. Ukuhlaziywa kwesakhiwo sezakhi zofuzo kubonise ukuthi ama-64.7% ezakhi zofuzo ze-BjGRF aqukethe ama-exons angu-4, okubonisa ukuthi isakhiwo sezakhi zofuzo ze-BjGRF silondolozwe ngokuziphendukela kwemvelo, kodwa inani lama-exons kumazakhi zofuzo ze-BjGRF10, BjGRF16, BjGRP28 kanye ne-BjGRF29 likhulu. Izifundo zibonise ukuthi ukwengeza noma ukususa ama-exons noma ama-intron kungaholela ekwehlukeni kwesakhiwo nomsebenzi wezakhi zofuzo, ngaleyo ndlela kudale izakhi zofuzo ezintsha ezingu-34,35,36. Ngakho-ke, sicabanga ukuthi i-intron ye-BjGRF yalahleka ngesikhathi sokuziphendukela kwemvelo, okungase kubangele izinguquko emsebenzini wezakhi zofuzo. Ngokuhambisana nezifundo ezikhona, sithole nokuthi inani lama-intron lalihlotshaniswa nokubonakaliswa kwezakhi zofuzo. Lapho inani lama-intron ku-gene likhulu, i-gene ingasabela ngokushesha ezicini ezahlukene ezingezinhle.
Ukuphindaphindwa kwezakhi zofuzo kuyisici esiyinhloko ekuziphendukeleni kwezakhi zofuzo kanye nezakhi zofuzo37. Izifundo ezihlobene zibonise ukuthi ukuphindaphindwa kwezakhi zofuzo akugcini nje ngokwandisa inani lezakhi zofuzo ze-GRF, kodwa futhi kusebenza njengendlela yokukhiqiza izakhi zofuzo ezintsha ukusiza izitshalo ukuba zivumelane nezimo ezahlukene zemvelo ezimbi38. Kutholakale izakhi zofuzo eziphindwe kabili ezingu-48 kulolu cwaningo, zonke ezaziyizakhi zofuzo eziphindaphindwayo, okubonisa ukuthi ukuphindaphindwa kwezakhi zofuzo kuyindlela enkulu yokwandisa inani lezakhi zofuzo kulo mndeni. Kubikwe ezincwadini ukuthi ukuphindaphindwa kwezakhi zofuzo kungakhuthaza ngempumelelo ukukhuliswa kwamalungu omndeni wezakhi zofuzo ze-GRF ku-Arabidopsis kanye ne-strawberry, futhi akukho ukuphindaphindwa kwezakhi zofuzo okutholakale kunoma yiluphi uhlobo27,39. Imiphumela yalolu cwaningo ihambisana nezifundo ezikhona ngemindeni ye-Arabidopsis thaliana kanye ne-strawberry, okuphakamisa ukuthi umndeni we-GRF ungandisa inani lezakhi zofuzo futhi ukhiqize izakhi zofuzo ezintsha ngokuphindaphindwa kwezakhi zofuzo ezitshalweni ezahlukene.
Kulolu cwaningo, kutholakale izakhi zofuzo ze-BjGRF ezingu-34 kumasipi, ezahlukaniswa zaba yimindeni emincane emi-3. Lezi zakhi zofuzo zibonise ama-motifs afanayo agciniwe kanye nezakhiwo zezakhi zofuzo. Ukuhlaziywa kwe-Collinearity kwembule ama-pair angu-48 okuphindaphindwa kwezingxenye kumasipi. Isifunda se-BjGRF promoter siqukethe izakhi ezisebenza nge-cis ezihlotshaniswa nokuphendula kokukhanya, impendulo yamahomoni, impendulo yokucindezeleka kwemvelo, kanye nokukhula nentuthuko. Ukuvezwa kwezakhi zofuzo ze-BjGRF ezingu-34 kutholakale esigabeni sokutshala kwesithombo semasipi (izimpande, iziqu, amaqabunga), kanye nephethini yokubonakaliswa kwezakhi zofuzo ze-BjGRF ezingu-10 ngaphansi kwezimo zesomiso. Kutholakale ukuthi amaphethini okubonakaliswa kwezakhi zofuzo ze-BjGRF ngaphansi kokucindezeleka kwesomiso ayefana futhi angase afane. ukubandakanyeka kwesomiso Ukulawula okuphoqelela. Izakhi zofuzo ze-BjGRF03 kanye ne-BjGRF32 zingadlala indima enhle yokulawula ekucindezelekeni kwesomiso, kuyilapho i-BjGRF06 kanye ne-BjGRF23 zidlala indima ekucindezelekeni kwesomiso njengezakhi zofuzo eziqondiwe ze-miR396. Sekukonke, ucwaningo lwethu luhlinzeka ngesisekelo sebhayoloji sokutholwa kwesikhathi esizayo komsebenzi wezakhi zofuzo ze-BjGRF ezitshalweni ze-Brassicaceae.
Imbewu yesinaphi esetshenziswe kulolu vivinyo inikezwe yiGuizhou Oil Seed Research Institute, iGuizhou Academy of Agricultural Sciences. Khetha imbewu yonke bese uyitshala enhlabathini (isisekelo: inhlabathi = 3:1), bese uqoqa izimpande, iziqu namaqabunga ngemuva kwesigaba samaqabunga amane. Izitshalo zaphathwa nge-20% PEG 6000 ukuze zilingise isomiso, futhi amaqabunga aqoqwa ngemva kwamahora angu-0, 3, 6, 12 kanye nama-24. Wonke amasampula ezitshalo aqandisiwe ngokushesha ku-nitrogen ewuketshezi abese egcinwa efrijini elingu--80°C ukuze kuhlolwe okulandelayo.
Yonke idatha etholwe noma ehlaziywe phakathi nalolu cwaningo ifakiwe esihlokweni esishicilelwe kanye namafayela olwazi olwengeziwe.
Isikhathi sokuthunyelwe: Jan-22-2025