Ukusasazwa kwamaxesha athile emvula kwiPhondo laseGuizhou akulingani, kukho imvula engakumbi entwasahlobo nasehlotyeni, kodwa izithole zembewu yedlwengulo zisesichengeni soxinzelelo lwembalela ekwindla nasebusika, nto leyo echaphazela kakhulu isivuno. Imostade sisityalo esikhethekileyo esilinywa kwiPhondo laseGuizhou. Iyakwazi ukumelana nembalela kwaye ingakhuliswa kwiindawo ezisezintabeni. Bubutyebi obuninzi bemizila yemfuza ekwaziyo ukumelana nembalela. Ukufunyanwa kwemfuza ekwazi ukumelana nembalela kubaluleke kakhulu ekuphuculweni kweentlobo zemostade. kunye nokutsha kwimithombo ye-germplasm. Usapho lweGRF ludlala indima ebalulekileyo ekukhuleni nasekuphuhliseni izityalo kunye nokusabela kuxinzelelo lwembalela. Okwangoku, iijini ze-GRF zifunyenwe kwi-Arabidopsis 2, irayisi (Oryza sativa) 12, i-rapeseed 13, umqhaphu (Gossypium hirsutum) 14, ingqolowa (Triticum). aestivum)15, pearl millet (Setaria italica)16 kunye neBrassica17, kodwa akukho zingxelo zofuzo zeGRF ezifunyenwe kwimostade. Kule sifundo, i-GRF yentsapho ye-mustard ye-mustard yachongwa kwinqanaba le-genome-wide kunye neempawu zabo zomzimba kunye neekhemikhali, ubudlelwane be-evolutionary, i-homology, i-motifs egciniweyo, i-gene structure, i-gene duplications, i-cis-elements kunye nesigaba se-seedling (isigaba seqabunga elinesine) zahlaziywa. Iipateni zokuthetha phantsi koxinzelelo lwembalela zahlalutywa ngokubanzi ukunika isiseko senzululwazi sokuqhubela phambili izifundo malunga nomsebenzi onokubakho wemfuza ye-BjGRF ekuphenduleni imbalela kunye nokubonelela ngemizila yemfuza yokuzala imostade ekwaziyo ukumelana nembalela.
Iintlobo ezingamashumi amathathu anesine ze-BjGRF zichongiwe kwi-Brassica juncea genome kusetyenziswa uphendlo olubini lwe-HMMER, zonke eziqulethe imimandla ye-QLQ kunye ne-WRC. Ulandelelwano lwe-CDS lweentlobo zemfuza ze-BjGRF ezichongiweyo zithiwe thaca kwiTheyibhile eyoNgezelelweyo S1. I-BjGRF01–BjGRF34 zithiywe ngokusekelwe kwindawo yazo kwichromosome. Iimpawu ze-physicochemical zale ntsapho zibonisa ukuba ubude be-amino acid buguquguquka kakhulu, ukusuka kwi-261 aa (BjGRF19) ukuya kwi-905 aa (BjGRF28). I-isoelectric point ye-BjGRF isuka kwi-6.19 (BjGRF02) ukuya kwi-9.35 (BjGRF03) kunye ne-avareji ye-8.33, kunye ne-88.24% ye-BjGRF yiprotheni eyisiseko. Uluhlu oluqikelelweyo lwe-molecular weight ye-BjGRF luvela kwi-29.82 kDa (BjGRF19) ukuya kwi-102.90 kDa (BjGRF28); isalathisi sokungazinzi kweeprotheyini ze-BjGRF ukusuka kwi-51.13 (BjGRF08) ukuya kwi-78.24 (BjGRF19), zonke zikhulu kune-40, ebonisa ukuba i-fatty acid index isuka kwi-43.65 (BjGRF01) ukuya kwi-78.78 (BjGRF22), i-avareji ye-78. (BjGRF31) ukuya -0.45 (BjGRF22), zonke iiprotheyini ze-hydrophilic ze-BjGRF zinexabiso elibi le-GRAVY, elinokuthi libe ngenxa yokungabikho kwe-hydrophobicity ebangelwa yintsalela. I-subcellular localization prediction ibonise ukuba i-31 BjGRF i-proteins encoded inokuthi ifakwe kwi-nucleus, i-BjGRF04 inokuthi ifakwe kwi-peroxisomes, i-BjGRF25 inokuthi ifakwe kwi-cytoplasm, kwaye i-BjGRF28 ifakwe kwi-chloroplasts (Itheyibhile 1), ebonisa ukuba i-BjGRF idlala indima ebalulekileyo kwi-nucleus kunye ne-nucleus. into ekhutshelweyo.
Uhlalutyo lwe-Phylogenetic lweentsapho ze-GRF kwiintlobo ezahlukeneyo zinokunceda ukufunda imisebenzi yemfuza. Ngoko ke, ulandelelwano olupheleleyo lwe-amino acid ye-35 ye-rapeseed, i-16 yetheniphu, i-12 yerayisi, i-millet eyi-10 kunye ne-9 Arabidopsis GRFs zakhutshelwa kwaye umthi we-phylogenetic wakhiwa ngokusekelwe kwi-34 echongiweyo yemfuza ye-BjGRF (Fig. 1). Iintsapho ezintathu ezingaphantsi ziqulathe amanani ahlukeneyo amalungu; I-116 GRF TFs zahlulwe zaba ziintsapho ezintathu ezahlukeneyo (amaqela A~C), aqulathe ama-59 (50.86%), ama-34 (29.31%) nama-23 (19.83)% eGRF, ngokulandelelanayo. Phakathi kwabo, amalungu osapho lwe-BjGRF angama-34 athe saa kwiintsapho ezi-3: amalungu ali-13 kwiqela A (38.24%), amalungu ali-12 kwiqela B (35.29%) kunye namalungu ali-9 kwiqela C (26.47%). Kwinkqubo ye-polyploidization ye-mustard, inani le-BjGRFs yofuzo kwii-subfamilies ezahlukeneyo zihluke, kwaye ukukhulisa i-gene kunye nokulahlekelwa kungenzeka. Kuyaphawuleka ukuba akukho ukuhanjiswa kwerayisi kunye ne-millet GRFs kwiqela C, ngelixa kukho i-2 yerayisi ye-GRF kunye ne-1 millet ye-GRF kwiqela B, kwaye uninzi lwee-GRF zerayisi kunye ne-millet zihlanganiswe kwisebe elinye, ebonisa ukuba i-BjGRF ihlobene ngokusondeleyo kwii-dicot. Phakathi kwazo, ezona zifundo zinzulu kakhulu ngomsebenzi we-GRF kwi-Arabidopsis thaliana zibonelela ngesiseko sezifundo ezisebenzayo ze-BjGRFs.
Umthi wePhylogenetic wemostade uquka iBrassica napus, iBrassica napus, irayisi, i-millet kunye namalungu osapho lwe-Arabidopsis thaliana GRF.
Uhlalutyo lwezakhi zofuzo eziphindaphindiweyo kusapho lwemostade GRF. Umgca ongwevu ngasemva ubonisa ibhloko ehambelanayo kwi-mustard genome, umgca obomvu ubonisa ukuphindaphinda okuphindwe kabini kwi-gene ye-BjGRF;
Imbonakalo yofuzo ye-BjGRF phantsi koxinzelelo lwembalela kwinqanaba lesine legqabi. Idatha ye-qRT-PCR iboniswe kwiTheyibhile eyoNgezelelweyo ye-S5. Ukwahluka okubalulekileyo kwiidatha kubonakaliswa ngoonobumba abancinci.
Njengoko imozulu yehlabathi iqhubeka nokutshintsha, ukufunda indlela izityalo ezihlangabezana ngayo noxinzelelo lwembalela kunye nokuphucula iindlela zabo zokunyamezela kuye kwaba sisihloko sophando olushushu18. Emva kwesomiso, i-morphological structure, i-gene expression kunye neenkqubo ze-metabolic zezityalo ziya kutshintsha, ezinokukhokelela ekuphelisweni kwe-photosynthesis kunye nokuphazamiseka kwe-metabolic, okuchaphazela isivuno kunye nomgangatho wezityalo19,20,21. Xa izityalo zibona iimpawu zesomiso, zivelisa izithunywa zesibini ezifana ne-Ca2 + kunye ne-phosphatidylinositol, ukwandisa i-intracellular calcium ion concentration kwaye isebenze inethiwekhi yokulawula i-protein phosphorylation pathway22,23. Iprotheyini yokugqibela ekujoliswe kuyo ibandakanyeka ngokuthe ngqo ekukhuseleni kwamaselula okanye ilawula ukubonakaliswa kwejene loxinzelelo oluhambelanayo ngokusebenzisa i-TFs, ukuphucula ukunyamezela kwezityalo kuxinzelelo24,25. Ngoko ke, iiTFs zidlala indima ebalulekileyo ekuphenduleni uxinzelelo lwembalela. Ngokolandelelwano kunye neempawu ezibophelelayo ze-DNA zoxinzelelo lwembalela eziphendula iiTFs, iiTFs zinokwahlulwa zibe ziintsapho ezahlukeneyo, ezifana neGRF, ERF, MYB, WRKY kunye nezinye iintsapho26.
Usapho lwe-GRF yofuzo luhlobo lwe-TF ethile yesityalo edlala indima ebalulekileyo kwiinkalo ezahlukeneyo ezifana nokukhula, uphuhliso, ukuhanjiswa komqondiso kunye neempendulo zokukhusela izityalo27. Ukusukela oko kwafunyaniswa ijene ye-GRF yokuqala kwi-O. sativa28, zininzi nangakumbi iijini ze-GRF ezichongiwe kwiintlobo ezininzi zezityalo kwaye zibonakaliswe ukuba zichaphazela ukukhula kwezityalo, uphuhliso kunye nokusabela koxinzelelo8, 29, 30,31,32. Ngokupapashwa kolandelelwano lwe-Brassica juncea genome, ukuchongwa kwe-BjGRF yosapho lwemfuza kuye kwenzeka33. Kolu phononongo, iijini ezingama-34 ze-BjGRF zichongiwe kuyo yonke igenome yemostade kwaye yabizwa ngokuba yi-BjGRF01-BjGRF34 ngokusekwe kwindawo yabo yechromosomal. Zonke ziqulethe imimandla ye-QLQ egcinwe kakhulu kunye ne-WRC. Uhlalutyo lweempawu ze-physicochemical lubonise ukuba ukuhlukana kwamanani e-amino acid kunye nobunzima be-molecular ye-BjGRF proteins (ngaphandle kwe-BjGRF28) yayingabalulekanga, ebonisa ukuba amalungu entsapho ye-BjGRF anokuba nemisebenzi efanayo. Uhlalutyo lwesakhiwo se-Gene lubonise ukuba i-64.7% ye-gene ye-BjGRF iqulethe i-4 exons, ebonisa ukuba i-BjGRF ye-gene structure igcinwe ngokulinganayo kwi-evolution, kodwa inani le-exons kwi-BjGRF10, i-BjGRF16, i-BjGRP28 kunye ne-BjGRF29 i-genes inkulu. Uphononongo lubonise ukuba ukongezwa okanye ukucinywa kwee-exons okanye ii-introns kunokukhokelela ekuhlukeni kwesakhiwo sofuzo kunye nomsebenzi, ngaloo ndlela kudala i-genes34,35,36 entsha. Ngoko ke, sicinga ukuba i-intron ye-BjGRF yalahleka ngexesha lokuziphendukela kwemvelo, oku kunokubangela utshintsho kwi-gene function. Ngokuhambelana nezifundo ezikhoyo, siphinde safumanisa ukuba inani le-introns lidibene ne-gene expression. Xa inani le-introns kwi-gene likhulu, i-gene inokukhawuleza ukuphendula kwizinto ezahlukeneyo ezingathandekiyo.
Ukuphindaphindwa kwemfuza yeyona nto iphambili kwi-genomic kunye ne-genetic evolution37. Uphononongo olunxulumeneyo lubonise ukuba uphindaphindo lwemfuza alondisi nje kuphela inani lemizila yemfuza ye-GRF, kodwa lukwasebenza njengendlela yokuvelisa imizila yemfuza emitsha ukunceda izityalo ukuba ziqhelane neemeko ezahlukeneyo zokusingqongileyo38. I-48 iyonke izibini eziphindwe kabini zofuzo zifunyenwe kolu phononongo, zonke ezo beziphindaphinda ngokwezigaba, nto leyo ebonisa ukuba ukuphindaphindwa kwamacandelo yeyona ndlela yokwandisa inani lemfuza kule ntsapho. Kuye kwaxelwa kuncwadi ukuba ukuphindaphinda kwecandelo kunokukhuthaza ngokufanelekileyo ukukhulisa amalungu osapho lwe-GRF gene kwi-Arabidopsis kunye nequnube, kwaye akukho phindaphindo lwe-tandem yolu sapho lwemfuza lufunyenwe kuyo nayiphi na i-species27,39. Iziphumo zolu phononongo zihambelana nezifundo ezikhoyo kwi-Arabidopsis thaliana kunye neentsapho ze-strawberry, ezibonisa ukuba intsapho ye-GRF inokunyusa inani leejeni kwaye ivelise ijene elitsha ngokuphindaphinda kwecandelo kwizityalo ezahlukeneyo.
Kule sifundo, i-34 ye-BjGRF i-genes yachongwa kwi-mustard, eyahlulwa yaba yi-3 subfamilies. Ezi mfuza zibonisa iimotifs ezifanayo ezigciniweyo kunye nezakhiwo zemfuza. Uhlalutyo lwe-Collinearity lubonise izibini ezingama-48 zecandelo lokuphindaphinda kwimostade. Ummandla womgqugquzeli we-BjGRF uqulethe izinto ze-cis-acting ezinxulumene nempendulo yokukhanya, impendulo ye-hormonal, impendulo yoxinzelelo lwendalo, kunye nokukhula kunye nophuhliso. Ukubonakaliswa kwe-34 BjGRF yofuzo kwachongwa kwinqanaba lembewu yemostade (iingcambu, iziqu, amagqabi), kunye nepateni ye-10 BjGRF phantsi kweemeko zembalela. Kwafunyaniswa ukuba iipatheni zokuvakalisa iijene ze-BjGRF phantsi koxinzelelo lwembalela ziyafana kwaye ziyafana. ukubandakanyeka kwimbalela Ukunyanzelisa ukulawulwa. Iijini ze-BjGRF03 kunye ne-BjGRF32 zinokudlala indima yolawulo elungileyo kuxinzelelo lwembalela, ngelixa i-BjGRF06 kunye ne-BjGRF23 zidlala indima kuxinzelelo lwembalela njengemiR396 yemfuza ekujoliswe kuyo. Ngokubanzi, uphononongo lwethu lubonelela ngesiseko sebhayoloji sokufunyanwa kwexesha elizayo lomsebenzi wemfuza we-BjGRF kwizityalo zeBrassicaceae.
Imbewu yemostade esetyenzisiweyo kolu vavanyo ibonelelwe yiGuizhou Oil Seed Research Institute, iGuizhou Academy of Agricultural Sciences. Khetha imbewu yonke kwaye uyityale emhlabeni (substrate: umhlaba = 3:1), kwaye uqokelele iingcambu, iziqu kunye namagqabi emva kwesigaba samagqabi amane. Izityalo zaphathwa nge-20% PEG 6000 ukulinganisa isomiso, kwaye amaqabunga aqokelelwa emva kwe-0, 3, 6, 12 kunye ne-24 iiyure. Zonke iisampulu zezityalo zaye zafakwa emkhenkceni ngoko nangoko kulwelo lwenitrogen zaza zagcinwa kwisikhenkcisi esingama-80°C kuvavanyo olulandelayo.
Yonke idatha efunyenweyo okanye ehlalutyiweyo ngexesha lolu phononongo ifakwe kwinqaku elipapashwe kunye neefayile zolwazi ezongezelelweyo.
Ixesha lokuposa: Jan-22-2025