Ukusasazwa kwemvula ngexesha lonyaka kwiPhondo laseGuizhou akulunganga, kunemvula eninzi entwasahlobo nasehlotyeni, kodwa izithole zerapeseed zisengozini yoxinzelelo lwembalela ekwindla nasebusika, nto leyo echaphazela kakhulu isivuno. I-Mustard sisityalo esikhethekileyo sembewu yeoyile esikhula kakhulu kwiPhondo laseGuizhou. Sinokunyamezelana okukhulu nembalela kwaye sinokukhuliswa kwiindawo zeentaba. Sisixhobo esityebileyo seejini ezimelana nembalela. Ukufunyanwa kweejini ezimelana nembalela kubaluleke kakhulu ekuphuculeni iintlobo zemasitadhi. kunye nokuvelisa izinto ezintsha kwizixhobo ze-germplasm. Usapho lwe-GRF ludlala indima ebalulekileyo ekukhuleni nasekuphuhlisweni kwezityalo kunye nokuphendula kuxinzelelo lwembalela. Okwangoku, iijini ze-GRF zifunyenwe kwi-Arabidopsis 2, irayisi (Oryza sativa) 12, irapeseed 13, umqhaphu (Gossypium hirsutum) 14, ingqolowa (Triticum). aestivum)15, i-pearl millet (Setaria italica)16 kunye neBrassica17, kodwa akukho ngxelo zejini ze-GRF ezifunyenwe kwimasitadhi. Kolu phononongo, ii-genes zosapho lwe-GRF ze-mustard zichongiwe kwinqanaba le-genome ngokubanzi kwaye iimpawu zazo zomzimba nezekhemikhali, ubudlelwane bokuvela kwezinto, i-homology, ii-motifs ezigciniweyo, ulwakhiwo lwe-gene, ukuphindaphinda kwe-gene, izinto ze-cis kunye nesigaba sembewu (isigaba samagqabi amane) zihlalutywe. Iipateni zokubonakaliswa phantsi koxinzelelo lwembalela zihlalutywe ngokupheleleyo ukuze zibonelele ngesiseko sesayensi sezifundo ezongezelelweyo malunga nomsebenzi onokubakho we-genes ze-BjGRF ekuphenduleni imbalela kunye nokubonelela ngee-genes ezifanelekileyo zokuzala i-mustard enyamezela imbalela.
Iijini ze-BjGRF ezingamashumi amathathu anesine zichongiwe kwi-genome ye-Brassica juncea kusetyenziswa uphando olubini lwe-HMMER, zonke eziqulathe ii-domains ze-QLQ kunye ne-WRC. Ulandelelwano lwe-CDS lweejini ze-BjGRF ezichongiweyo luboniswe kwiTheyibhile eyoNgezelelweyo S1. I-BjGRF01–BjGRF34 zibizwa ngokusekwe kwindawo ezikuyo kwi-chromosome. Iipropati ze-physicochemical zale family zibonisa ukuba ubude be-amino acid buyahluka kakhulu, ukusuka kwi-261 aa (BjGRF19) ukuya kwi-905 aa (BjGRF28). I-isoelectric point ye-BjGRF isusela kwi-6.19 (BjGRF02) ukuya kwi-9.35 (BjGRF03) ngomyinge we-8.33, kwaye i-88.24% ye-BjGRF yiproteni esisiseko. Uluhlu oluqikelelweyo lobunzima be-molecular lwe-BjGRF luqala kwi-29.82 kDa (BjGRF19) ukuya kwi-102.90 kDa (BjGRF28); isalathisi sokungazinzi kweeproteni ze-BjGRF siqala kwi-51.13 (BjGRF08) ukuya kwi-78.24 (BjGRF19), zonke zingaphezulu kwe-40, nto leyo ebonisa ukuba isalathisi se-fatty acid siqala kwi-43.65 (BjGRF01) ukuya kwi-78.78 (BjGRF22), i-hydrophilicity ephakathi (GRAVY) isukela kwi--1.07 (BjGRF31) ukuya kwi--0.45 (BjGRF22), zonke iiproteni ze-BjGRF ezi-hydrophilic zinexabiso le-GRAVY elibi, elinokubangelwa kukungabikho kwe-hydrophobicity ebangelwa ziisalela. Uqikelelo lwendawo engaphantsi kweeseli lubonise ukuba iiproteni ezingama-31 ze-BjGRF ezifakwe kwi-nucleus zinokufunyanwa kwindawo ethile, i-BjGRF04 inokufunyanwa kwindawo ethile kwii-peroxisomes, i-BjGRF25 inokufunyanwa kwindawo ethile kwi-cytoplasm, kwaye i-BjGRF28 inokufunyanwa kwindawo ethile kwii-chloroplasts (Itheyibhile 1), nto leyo ebonisa ukuba ii-BjGRF zinokufunyanwa kwindawo ethile kwi-nucleus kwaye zidlale indima ebalulekileyo yokulawula njengento yokubhala.
Uhlalutyo lwe-phylogenetic lweentsapho ze-GRF kwiintlobo ezahlukeneyo lunokunceda ekufundeni imisebenzi ye-gene. Ke ngoko, ulandelelwano olupheleleyo lwe-amino acid lwe-35 rapeseed, i-turnip ezili-16, irayisi eli-12, i-millet eli-10 kunye ne-9 Arabidopsis GRF zakhutshelwa kwaye umthi we-phylogenetic wakhiwa ngokusekelwe kwii-genes ezingama-34 ze-BjGRF ezichongiweyo (Umzobo 1). Ezi ntsapho zintathu ziqulathe amanani ahlukeneyo amalungu; ii-TF ze-GRF ezili-116 zahlulwe zaba ziintsapho ezintathu ezahlukeneyo (amaqela A~C), aqulathe ama-59 (50.86%), ama-34 (29.31%) kunye nama-23 (19.83)% ee-GRF, ngokwahlukeneyo. Phakathi kwazo, amalungu osapho lwe-BjGRF angama-34 asasazeke kwiintsapho ezi-3 ezincinci: amalungu ali-13 kwiqela A (38.24%), amalungu ali-12 kwiqela B (35.29%) kunye namalungu ali-9 kwiqela C (26.47%). Kwinkqubo ye-mustard polyploidization, inani leejini ze-BjGRF kwiintsapho ezahlukeneyo zahlukile, kwaye ukwandiswa kunye nokulahleka kwejini kusenokwenzeka ukuba kwenzeke. Kubalulekile ukuqaphela ukuba akukho kusasazeka kwe-rayisi kunye ne-millet GRF kwiqela C, ngelixa kukho ii-rayisi ezimbini ze-GRF kunye ne-millet GRF e-1 kwiqela B, kwaye uninzi lwee-rayisi kunye ne-millet GRF zihlanganiswe kwisebe elinye, nto leyo ebonisa ukuba ii-BjGRF zinxulumene kakhulu nee-dicots. Phakathi kwazo, izifundo ezinzulu kakhulu ngomsebenzi we-GRF kwi-Arabidopsis thaliana zibonelela ngesiseko sezifundo ezisebenzayo ze-BjGRF.
Umthi we-mustard oquka iBrassica napus, iBrassica napus, irayisi, i-millet kunye namalungu osapho lwe-Arabidopsis thaliana GRF.
Uhlalutyo lweejini eziphindaphindayo kusapho lwe-mustard GRF. Umgca ongwevu ngasemva umele ibhloko ehambelanayo kwi-genome ye-mustard, umgca obomvu umele isibini seephindaphindo ezihlukeneyo ze-gene ye-BjGRF;
Ukubonakaliswa kwe-gene ye-BjGRF phantsi koxinzelelo lwembalela kwinqanaba lesine lamagqabi. Idatha ye-qRT-PCR iboniswe kwiTheyibhile eyoNgezelelweyo S5. Umahluko omkhulu kwidatha uboniswa ngoonobumba abancinci.
Njengoko imozulu yehlabathi iqhubeka nokutshintsha, ukufunda indlela izityalo ezihlangabezana ngayo noxinzelelo lwembalela kunye nokuphucula iindlela zazo zokunyamezelana kube ngumxholo oshushu wophando18. Emva kwembalela, ulwakhiwo lwe-morphological, ukubonakaliswa kwezakhi zofuzo kunye neenkqubo ze-metabolic zezityalo ziya kutshintsha, nto leyo enokukhokelela ekuphelisweni kwe-photosynthesis kunye nokuphazamiseka kwe-metabolic, okuchaphazela isivuno kunye nomgangatho wezityalo19,20,21. Xa izityalo zibona imiqondiso yembalela, zivelisa izithunywa zesibini ezifana ne-Ca2+ kunye ne-phosphatidylinositol, zonyusa uxinzelelo lwe-calcium ion ngaphakathi kweseli kwaye zisebenze inethiwekhi yolawulo yendlela ye-protein phosphorylation22,23. Iproteni yokugqibela ekujoliswe kuyo ibandakanyeka ngokuthe ngqo ekukhuseleni iiseli okanye ilawula ukubonakaliswa kwezakhi zofuzo ezinxulumeneyo ngee-TF, iphucula ukunyamezela kwezityalo kuxinzelelo24,25. Ke ngoko, ii-TF zidlala indima ebalulekileyo ekuphenduleni kuxinzelelo lwembalela. Ngokwendlela yokulandelelana kunye neempawu zokubopha i-DNA zee-TF ezisabela kuxinzelelo lwembalela, ii-TF zinokwahlulwa zibe ziintsapho ezahlukeneyo, ezifana ne-GRF, ERF, MYB, WRKY kunye nezinye iintsapho26.
Usapho lwe-GRF luhlobo lwe-TF ethile yesityalo edlala indima ebalulekileyo kwiinkalo ezahlukeneyo ezifana nokukhula, uphuhliso, ukudluliselwa kwesignali kunye neempendulo zokuzikhusela kwezityalo27. Ukususela oko i-gene yokuqala ye-GRF yachongwa kwi-O. sativa28, ii-genes ezininzi ze-GRF zichongiwe kwiintlobo ezininzi kwaye zibonakaliswe ukuba zichaphazela ukukhula kwezityalo, uphuhliso kunye nempendulo yoxinzelelo8, 29, 30,31,32. Ngokupapashwa kolandelelwano lwe-genome ye-Brassica juncea, ukuchongwa kosapho lwe-gene ye-BjGRF kwaba nokwenzeka33. Kolu phononongo, ii-genes ze-BjGRF ezingama-34 zachongwa kwi-genome ye-mustard yonke kwaye zabizwa ngokuba yi-BjGRF01–BjGRF34 ngokusekelwe kwindawo yazo ye-chromosome. Zonke ziqulathe ii-domains ze-QLQ kunye ne-WRC ezigcinwe kakhulu. Uhlalutyo lweempawu ze-physicochemical lubonise ukuba umahluko kwiinombolo ze-amino acid kunye nobunzima beemolekyuli zeeproteni ze-BjGRF (ngaphandle kwe-BjGRF28) azibalulekanga, nto leyo ebonisa ukuba amalungu osapho lwe-BjGRF anokuba nemisebenzi efanayo. Uhlalutyo lwesakhiwo sejini lubonise ukuba i-64.7% yejini ze-BjGRF ziqulathe ii-exons ezi-4, nto leyo ebonisa ukuba isakhiwo sejini ze-BjGRF sigcinwe kakuhle kwindalo, kodwa inani lee-exons kwijini ze-BjGRF10, BjGRF16, BjGRP28 kunye ne-BjGRF29 likhulu. Izifundo zibonise ukuba ukongezwa okanye ukususwa kwee-exons okanye ii-intron kunokukhokelela kumahluko kwisakhiwo sejini kunye nomsebenzi, ngaloo ndlela kudala iijini ezintsha34,35,36. Ke ngoko, sicinga ukuba i-intron ye-BjGRF yalahleka ngexesha lokuvela, nto leyo enokubangela utshintsho kumsebenzi wejini. Ngokuhambelana nezifundo ezikhoyo, sikwafumanise ukuba inani lee-introns lalinxulunyaniswa nokubonakaliswa kwejini. Xa inani lee-introns kwijini likhulu, ijini inokuphendula ngokukhawuleza kwizinto ezahlukeneyo ezingalunganga.
Ukuphindaphinda kwezakhi zofuzo yinto ephambili ekuguqukeni kwezakhi zofuzo kunye nezakhi zofuzo37. Izifundo ezinxulumene noko zibonise ukuba ukuphindaphinda kwezakhi zofuzo akukhulisi nje kuphela inani lezakhi zofuzo ze-GRF, kodwa kukwasebenza njengendlela yokuvelisa izakhi zofuzo ezintsha ukunceda izityalo zilungelelanise neemeko ezahlukeneyo zokusingqongileyo ezingalunganga38. Kufunyenwe izibini zezakhi zofuzo eziphindwe kabini ezingama-48 kolu phononongo, zonke ezazizii-segmental duplications, nto leyo ebonisa ukuba ukuphindaphinda kwezakhi zofuzo yindlela ephambili yokwandisa inani lezakhi zofuzo kolu sapho. Kuye kwaxelwa kwiincwadi ukuba ukuphindaphinda kwezakhi zofuzo kunokukhuthaza ngempumelelo ukwanda kwamalungu osapho lwezakhi zofuzo ze-GRF kwi-Arabidopsis kunye ne-strawberry, kwaye akukho kuphinda kabini kwale ntsapho yezakhi zofuzo okufunyenweyo kuyo nayiphi na intlobo27,39. Iziphumo zolu phononongo ziyahambelana nezifundo ezikhoyo kwiintsapho ze-Arabidopsis thaliana kunye ne-strawberry, nto leyo ebonisa ukuba usapho lwe-GRF lunokwandisa inani lezakhi zofuzo kwaye luvelise izakhi zofuzo ezintsha ngokuphindaphindwa kwezakhi zofuzo kwizityalo ezahlukeneyo.
Kolu phononongo, ii-genes ze-BjGRF ezingama-34 zichongiwe kwi-mustard, ezahlulwe zaba ziintsapho ezi-3 ezincinci. Ezi genes zibonise ii-motifs ezifanayo ezigciniweyo kunye nezakhiwo ze-gene. Uhlalutyo lwe-Collinearity lutyhile ii-segment duplications ezingama-48 kwi-mustard. Ummandla we-BjGRF promoter uqulethe izinto ezisebenza nge-cis ezinxulumene nokuphendula kokukhanya, impendulo ye-hormone, impendulo yoxinzelelo lokusingqongileyo, kunye nokukhula kunye nophuhliso. Ukubonakaliswa kwee-genes ze-BjGRF ezingama-34 kufunyenwe kwinqanaba le-mustard seedling (iingcambu, iziqu, amagqabi), kunye nephethini yokubonakaliswa kwee-genes ze-BjGRF ezili-10 phantsi kweemeko zembalela. Kufunyenwe ukuba iipatheni zokubonakaliswa kwee-genes ze-BjGRF phantsi koxinzelelo lwembalela zazifana kwaye zinokufana. ukubandakanyeka kwimbalela. Ulawulo olunyanzelisayo. Ii-genes ze-BjGRF03 kunye ne-BjGRF32 zinokudlala indima elungileyo yokulawula kwimbalela, ngelixa i-BjGRF06 kunye ne-BjGRF23 zidlala indima kwimbalela njengee-genes ezijoliswe kwi-miR396. Ngokubanzi, uphando lwethu lubonelela ngesiseko sebhayoloji sokufunyanwa kwexesha elizayo komsebenzi we-gene ye-BjGRF kwizityalo zaseBrassicaceae.
Imbewu yemostade esetyenziswe kolu vavanyo inikezelwe yiGuizhou Oil Seed Research Institute, iGuizhou Academy of Agricultural Sciences. Khetha imbewu epheleleyo uze uyityale emhlabeni (umhlaba ongaphantsi komhlaba = 3:1), uze uqokelele iingcambu, iziqu kunye namagqabi emva kwesigaba samagqabi amane. Izityalo ziphathwe nge-20% PEG 6000 ukuze zilinganise imbalela, kwaye amagqabi aqokelelwa emva kweeyure eziyi-0, 3, 6, 12 kunye nama-24. Zonke iisampulu zezityalo zaqandiselwa ngoko nangoko kwi-nitrogen engamanzi zaze zagcinwa kwifriji ye--80°C kuvavanyo olulandelayo.
Lonke ulwazi olufunyenweyo okanye oluhlalutyiweyo ngeli xesha lophando lufakiwe kwinqaku elishicilelweyo kunye neefayile zolwazi olongezelelweyo.
Ixesha leposi: Jan-22-2025