Ukusatshalaliswa kwezinkathi ezithile zemvula eSifundazweni saseGuizhou akulingani, kunemvula eyengeziwe entwasahlobo nasehlobo, kodwa izithombo ze-rapeseed zisengozini yesomiso ekwindla nasebusika, okuthinta kakhulu isivuno. I-mustard isitshalo esikhethekile sembewu kawoyela etshalwa kakhulu esifundazweni saseGuizhou. Iyakwazi ukumelana nesomiso futhi ingatshalwa ezindaweni ezinezintaba. Iwumthombo ocebile wezakhi zofuzo ezimelana nesomiso. Ukutholakala kwezakhi zofuzo ezimelana nesomiso kubaluleke kakhulu ekwenzeni ngcono izinhlobo zesinaphi. kanye nokusungulwa kwezinsiza ze-germplasm. Umndeni we-GRF udlala indima ebalulekile ekukhuleni nasekuthuthukeni kwezitshalo kanye nokusabela ekucindezelekeni kwesomiso. Njengamanje, izakhi zofuzo ze-GRF zitholakele ku-Arabidopsis 2, irayisi (Oryza sativa) 12, i-rapeseed 13, ukotini (Gossypium hirsutum) 14, ukolweni (Triticum). aestivum)15, pearl millet (Setaria italica)16 kanye ne-Brassica17, kodwa ayikho imibiko yezakhi zofuzo ze-GRF ezitholwe ku-mustard. Kulolu cwaningo, izakhi zofuzo zomndeni we-GRF zesinaphi zihlonzwe ezingeni elibanzi le-genome kanye nezici zabo zomzimba kanye namakhemikhali, ubudlelwano bokuziphendukela kwemvelo, i-homology, i-motifs elondoloziwe, isakhiwo sofuzo, ukuphindaphinda kwezakhi zofuzo, i-cis-elements kanye nesiteji sezithombo (isigaba samaqabunga amane) kwahlaziywa. Amaphethini enkulumo ngaphansi kwengcindezi yesomiso ahlaziywa kabanzi ukuze kuhlinzekwe isisekelo sesayensi socwaningo olwengeziwe mayelana nokusebenza okungaba khona kwezakhi zofuzo ze-BjGRF ekubhekaneni nesomiso kanye nokuhlinzeka ngezakhi zofuzo zokuzalanisa lwesinaphi esikwazi ukumelana nesomiso.
Izakhi zofuzo ze-BjGRF ezingamashumi amathathu nane zihlonzwe ku-Brassica juncea genome kusetshenziswa ukusesha okubili kwe-HMMER, konke okuqukethe izizinda ze-QLQ ne-WRC. Ukulandelana kwe-CDS kwezakhi zofuzo ze-BjGRF ezihlonziwe zethulwa ku-Supplementary Table S1. I-BjGRF01–BjGRF34 ziqanjwe ngokusekelwe endaweni yazo ku-chromosome. Izici ze-physicochemical zalo mndeni zibonisa ukuthi ubude be-amino acid buyahlukahluka kakhulu, kusukela ku-261 aa (BjGRF19) kuya ku-905 aa (BjGRF28). Iphoyinti le-isoelectric le-BjGRF lisukela ku-6.19 (BjGRF02) kuya ku-9.35 (BjGRF03) ngesilinganiso esingu-8.33, kanti u-88.24% we-BjGRF iphrotheni eyisisekelo. Ibanga lesisindo samangqamuzana elibikezelwe le-BjGRF lisuka ku-29.82 kDa (BjGRF19) ukuya ku-102.90 kDa (BjGRF28); inkomba yokungaqini kwamaprotheni e-BjGRF isuka ku-51.13 (BjGRF08) kuya ku-78.24 (BjGRF19), wonke makhulu kuno-40, okubonisa ukuthi inkomba ye-fatty acid isuka ku-43.65 (BjGRF01) iye ku-78.78 (BjGRF22), i-avareji yebanga le-700000000000000000000000, ukusuka ku-78. (BjGRF31) kuya -0.45 (BjGRF22), wonke amaprotheni e-BjGRF angama-hydrophilic anamanani angemahle we-GRAVY, okungenzeka kungenxa yokuntuleka kwe-hydrophobicity okubangelwa izinsalela. Isibikezelo sendawo engaphansi kwamaselula sibonise ukuthi amaprotheni angu-31 BjGRF afakwe ikhodi angenziwa endaweni ku-nucleus, i-BjGRF04 ingenziwa endaweni kuma-peroxisomes, i-BjGRF25 ingenziwa endaweni ku-cytoplasm, futhi i-BjGRF28 ingenziwa endaweni kuma-chloroplasts (Ithebula 1), okubonisa ukuthi indima ye-BjGRF yendawo ebalulekile ingase idlale indima ebalulekile njenge-nucleus isici sokuloba.
Ukuhlaziywa kwe-Phylogenetic kwemindeni ye-GRF ezinhlotsheni ezahlukene kungasiza ukutadisha imisebenzi yofuzo. Ngakho-ke, ukulandelana kobude obugcwele be-amino acid ye-rapeseed engu-35, itheniphu engu-16, irayisi engu-12, i-millet engu-10 kanye ne-9 Arabidopsis GRFs yalandwa futhi isihlahla se-phylogenetic sakhiwa ngokusekelwe kuzakhi zofuzo ze-BjGRF ezingu-34 (Fig. 1). Imindeni emithathu engaphansi iqukethe izinombolo ezihlukene zamalungu; I-116 GRF TF ihlukaniswe yaba imindeni emithathu ehlukene (amaqembu A~C), aqukethe ama-59 (50.86%), 34 (29.31%) kanye nama-23 (19.83)% ama-GRF, ngokulandelana. Phakathi kwabo, amalungu omndeni we-BjGRF angu-34 ahlakazeke emindenini emi-3 emincane: amalungu angu-13 eqenjini A (38.24%), amalungu angu-12 eqenjini B (35.29%) namalungu angu-9 eqenjini C (26.47%). Enqubweni ye-polyploidization yesinaphi, inani lezakhi zofuzo ze-BjGRFs emindenini engaphansi ehlukene lihlukile, futhi ukukhuliswa kofuzo nokulahlekelwa kungenzeka. Kuyaphawuleka ukuthi akukho ukusatshalaliswa kwerayisi ne-millet GRFs eqenjini C, kuyilapho kukhona 2 rice GRFs kanye 1 millet GRF eqenjini B, futhi iningi rice kanye namabele GRFs kuhlanganiswe egatsheni elilodwa, okubonisa ukuthi BjGRFs ahlobene eduze dicots. Phakathi kwazo, izifundo ezijule kakhulu ngomsebenzi we-GRF ku-Arabidopsis thaliana zinikeza isisekelo sezifundo ezisebenzayo zama-BjGRF.
Isihlahla se-Phylogenetic sesinaphi esihlanganisa i-Brassica napus, i-Brassica napus, irayisi, amabele kanye namalungu omndeni we-Arabidopsis thaliana GRF.
Ukuhlaziywa kwezakhi zofuzo eziphindaphindayo emndenini we-mustard GRF. Umugqa ompunga ngemuva umelela ibhulokhi evumelanisiwe kufuzo lwesinaphi, umugqa obomvu umelela ukuphindaphinda okuhlukaniswe ngamasegimenti kofuzo lwe-BjGRF;
I-BjGRF gene expression ngaphansi kwengcindezi yesomiso esigabeni sesine seqabunga. Idatha ye-qRT-PCR ikhonjiswe kokuthi Ithebula Le-Supplementary S5. Umehluko omkhulu kudatha uboniswa ngezinhlamvu ezincane.
Njengoba isimo sezulu somhlaba siqhubeka nokushintsha, ukufunda ukuthi izitshalo zibhekana kanjani nengcindezi yesomiso kanye nokwenza ngcono izindlela zazo zokubekezelela sekuyisihloko socwaningo esishisayo18. Ngemuva kwesomiso, ukwakheka kwe-morphological, i-gene expression kanye nezinqubo ze-metabolic zezitshalo zizoshintsha, okungase kuholele ekumisweni kwe-photosynthesis nokuphazamiseka kwe-metabolic, okuthinta isivuno kanye nekhwalithi yezitshalo19,20,21. Lapho izitshalo zizwa izimpawu zesomiso, zikhiqiza izithunywa zesibili ezifana ne-Ca2 + ne-phosphatidylinositol, zandise ukugxila kwe-calcium ion ye-intracellular futhi zisebenze inethiwekhi yokulawula ye-protein phosphorylation pathway22,23. Iprotheyini yokugcina ehlosiwe ihileleke ngokuqondile ekuvikeleni amaselula noma ilawula ukubonakaliswa kwezakhi zofuzo ezihlobene nokucindezeleka ngokusebenzisa ama-TF, okuthuthukisa ukubekezelelana kwezitshalo ekucindezelekeni24,25. Ngakho, ama-TF adlala indima ebalulekile ekuphenduleni ingcindezi yesomiso. Ngokulandelana kanye nezakhiwo ezibophezelayo ze-DNA zama-TF asabelayo ekucindezelekeni kwesomiso, ama-TF angahlukaniswa abe imindeni ehlukene, njenge-GRF, ERF, MYB, WRKY kanye neminye imindeni26.
Umndeni we-GRF wofuzo uwuhlobo lwe-TF eqondene nesitshalo edlala izindima ezibalulekile ezicini ezihlukahlukene ezifana nokukhula, ukuthuthukiswa, ukudluliselwa kwesignali kanye nezimpendulo zokuvikela izitshalo27. Kusukela kutholwe isakhi sofuzo sokuqala se-GRF ku-O. sativa28, izakhi zofuzo ezengeziwe ze-GRF zikhonjwe ezinhlobonhlobo eziningi futhi ziboniswa ukuthi zithinta ukukhula kwezitshalo, ukukhula kanye nokusabela kwengcindezi8, 29, 30,31,32. Ngokushicilelwa kokulandelana kofuzo lwe-Brassica juncea, ukuhlonzwa komndeni wofuzo lwe-BjGRF kwaba nokwenzeka33. Kulolu cwaningo, izakhi zofuzo ezingu-34 ze-BjGRF zihlonzwe kulo lonke i-mustard genome futhi zaqanjwa ngokuthi BjGRF01–BjGRF34 ngokusekelwe endaweni yazo ye-chromosomal. Zonke ziqukethe izizinda ze-QLQ ne-WRC ezigcinwe kakhulu. Ukuhlaziywa kwezakhiwo ze-physicochemical kubonise ukuthi umehluko wezinombolo ze-amino acid nezisindo zamangqamuzana amaprotheni e-BjGRF (ngaphandle kwe-BjGRF28) ayengabalulekile, okubonisa ukuthi amalungu omndeni we-BjGRF angase abe nemisebenzi efanayo. Ukuhlaziywa kwesakhiwo sofuzo kubonise ukuthi u-64.7% wezakhi zofuzo ze-BjGRF ziqukethe ama-exons angu-4, okubonisa ukuthi isakhiwo sofuzo se-BjGRF sigcinwe ngokuqhathaniswa ekuziphendukeleni kwemvelo, kodwa inani lama-exons kuzakhi zofuzo ze-BjGRF10, BjGRF16, BjGRP28 kanye ne-BjGRF29 likhulu. Ucwaningo luye lwabonisa ukuthi ukungezwa noma ukususwa kwama-exons noma ama-intron kungaholela ekuhlukeni kwesakhiwo sofuzo kanye nokusebenza, ngaleyo ndlela kudala izakhi zofuzo ezintsha34,35,36. Ngakho-ke, sicabanga ukuthi i-intron ye-BjGRF yalahleka ngesikhathi sokuziphendukela kwemvelo, okungase kubangele izinguquko ekusebenzeni kwezakhi zofuzo. Ngokuhambisana nezifundo ezikhona, siphinde sathola ukuthi inani lama-intron lalihlotshaniswa nenkulumo yofuzo. Uma inani lama-intron esakhini likhulu, isakhi sofuzo singasabela ngokushesha ezicini ezihlukahlukene ezingezinhle.
Ukuphindaphinda izakhi zofuzo kuyisici esikhulu ekuguqukeni kofuzo kanye nezakhi zofuzo37. Ucwaningo oluhlobene lubonise ukuthi ukuphindaphinda kwezakhi zofuzo akukhulisi nje kuphela inani lezakhi zofuzo ze-GRF, kodwa futhi kusebenza njengendlela yokukhiqiza izakhi zofuzo ezintsha ukuze kusize izitshalo ukuthi zivumelane nezimo ezihlukahlukene ezimbi zemvelo38. Ingqikithi yamapheya ofuzo ayimpinda angama-48 atholwe kulolu cwaningo, konke obekuwukuphindaphindeka kwezigaba, okubonisa ukuthi ukuphindaphinda kwezigaba kuyindlela enkulu yokwandisa inani lezakhi zofuzo kulo mndeni. Kubikwe emibhalweni ukuthi ukuphindaphinda kwezigaba kungakhuthaza ngempumelelo ukukhuliswa kwamalungu omndeni we-GRF e-Arabidopsis kanye ne-strawberry, futhi akukho ukuphindaphinda kwe-tandem kwalo mndeni wofuzo okutholakele kunoma yiluphi uhlobo lwezinhlobo27,39. Imiphumela yalolu cwaningo iyahambisana nezifundo ezikhona ngemindeni ye-Arabidopsis thaliana kanye ne-strawberry, ephakamisa ukuthi umndeni we-GRF ungandisa inani lezakhi zofuzo futhi ukhiqize izakhi zofuzo ezintsha ngokuphindaphinda izingxenye ezitshalweni ezahlukene.
Kulolu cwaningo, ingqikithi yezakhi zofuzo ze-BjGRF ze-34 zihlonzwe ku-mustard, ezahlukaniswa zaba imindeni emincane emi-3. Lezi zakhi zofuzo zibonise i-motif efanayo elondoloziwe kanye nezakhi zofuzo. Ukuhlaziywa kwe-Collinearity kwembule amapheya angama-48 ezigaba eziphindaphindekayo zesinaphi. Isifunda somgqugquzeli we-BjGRF siqukethe izakhi ze-cis-acting ezihambisana nokusabela okukhanyayo, impendulo yehomoni, impendulo yokucindezeleka kwemvelo, nokukhula nokuthuthuka. Ukuvezwa kwezakhi zofuzo ezingu-34 ze-BjGRF kutholwe esigabeni sesithombo sesinaphi (izimpande, iziqu, amaqabunga), kanye nephethini yokukhuluma yezakhi zofuzo ezingu-10 ze-BjGRF ngaphansi kwezimo zesomiso. Kutholwe ukuthi amaphethini wokukhuluma wezakhi zofuzo ze-BjGRF ngaphansi kwengcindezi yesomiso ayefana futhi angase afane. ukuzibandakanya nesomiso Ukuphoqelela ukulawulwa. Izakhi zofuzo ze-BjGRF03 kanye ne-BjGRF32 zingadlala izindima ezinhle zokulawula ekucindezelekeni kwesomiso, kuyilapho i-BjGRF06 ne-BjGRF23 zidlala indima ekucindezelekeni kwesomiso njengezakhi zofuzo ezihlosiwe ze-miR396. Sekukonke, ucwaningo lwethu luhlinzeka ngesisekelo sebhayoloji sokutholwa kwesikhathi esizayo komsebenzi wofuzo we-BjGRF ezitshalweni ze-Brassicaceae.
Imbewu yesinaphi esetshenziswe kulokhu kuhlola inikezwe i-Guizhou Oil Seed Research Institute, i-Guizhou Academy of Agricultural Sciences. Khetha imbewu yonke bese uyitshala emhlabathini (i-substrate: inhlabathi = 3:1), bese uqoqa izimpande, iziqu namaqabunga emva kwesigaba samaqabunga amane. Izitshalo zaphathwa nge-20% PEG 6000 ukulingisa isomiso, futhi amaqabunga aqoqwa ngemva kwamahora angu-0, 3, 6, 12 kanye nama-24. Wonke amasampula ezitshalo aqandiswa ngokushesha ku-nitrogen ewuketshezi bese egcinwa efrijini elingu -80°C ukuze kuhlolwe okulandelayo.
Yonke idatha etholiwe noma ehlaziywe phakathi nalolu cwaningo ifakiwe esihlokweni esishicilelwe kanye namafayela olwazi olwengeziwe.
Isikhathi sokuthumela: Jan-22-2025