Kev faib tawm raws caij nyoog ntawm nag lossis daus hauv Guizhou xeev tsis sib npaug, nrog nag lossis daus ntau nyob rau lub caij nplooj ntoo hlav thiab lub caij ntuj sov, tab sis cov rapeseed yub tau raug cuam tshuam rau kev ntxhov siab hauv lub caij nplooj zeeg thiab lub caij ntuj no, uas cuam tshuam rau cov qoob loo. Mustard yog cov qoob loo tshwj xeeb oilseed feem ntau loj hlob hauv Guizhou Xeev. Nws muaj zog drought kam rau ua thiab tuaj yeem loj hlob hauv thaj chaw roob. Nws yog ib qho chaw nplua nuj ntawm drought-resistant noob. Kev tshawb pom ntawm cov noob qoob loo-resistant yog qhov tseem ceeb rau kev txhim kho cov noob txiv ntoo. thiab kev tsim kho tshiab hauv cov peev txheej kab mob. GRF tsev neeg ua lub luag haujlwm tseem ceeb hauv kev loj hlob thiab kev loj hlob ntawm cov nroj tsuag thiab cov lus teb rau kev ntxhov siab vim kev nyuaj siab. Tam sim no, GRF noob tau pom nyob rau hauv Arabidopsis 2, mov (Oryza sativa) 12, rapeseed 13, paj rwb (Gossypium hirsutum) 14, nplej (Triticum). aestivum)15, pearl millet (Setaria italica)16 thiab Brassica17, tab sis tsis muaj ntawv ceeb toom ntawm GRF genes kuaj pom hauv mustard. Hauv txoj kev tshawb no, GRF tsev neeg cov noob ntawm mustard tau txheeb xyuas ntawm qib genome-wide thiab lawv lub cev thiab tshuaj lom neeg cov yam ntxwv, evolutionary kev sib raug zoo, homology, conserved motifs, gene qauv, gene duplications, cis-cov ntsiab lus thiab sedling theem (plaub-nplooj theem) tau soj ntsuam. Cov qauv qhia nyob rau hauv drought kev nyuaj siab tau raug soj ntsuam los muab ib tug scientific lub hauv paus rau kev tshawb fawb ntxiv txog lub peev xwm ntawm BjGRF noob nyob rau hauv drought teb thiab muab cov neeg sib tw genes rau yug me nyuam drought-tolerant mustard.
Peb caug-plaub BjGRF noob tau txheeb xyuas hauv Brassica juncea genome siv ob qhov kev tshawb nrhiav HMMER, tag nrho cov uas muaj cov QLQ thiab WRC domains. CDS cov kab ke ntawm BjGRF cov noob tau txheeb xyuas tau nthuav tawm hauv Cov Lus Ntxiv S1. BjGRF01–BjGRF34 muaj npe raws li lawv qhov chaw nyob ntawm chromosome. Lub physicochemical zog ntawm tsev neeg no qhia tau hais tias cov amino acid ntev yog qhov sib txawv heev, xws li 261 aa (BjGRF19) txog 905 aa (BjGRF28). Lub isoelectric point ntawm BjGRF yog li ntawm 6.19 (BjGRF02) mus rau 9.35 (BjGRF03) nrog qhov nruab nrab ntawm 8.33, thiab 88.24% ntawm BjGRF yog ib qho yooj yim protein. Qhov kwv yees molecular hnyav ntau ntawm BjGRF yog los ntawm 29.82 kDa (BjGRF19) txog 102.90 kDa (BjGRF28); qhov instability Performance index ntawm BjGRF proteins ranges ntawm 51.13 (BjGRF08) mus rau 78.24 (BjGRF19), tag nrho cov yog ntau tshaj 40, qhia hais tias cov fatty acid index yog li ntawm 43.65 (BjGRF01) mus rau 78.78 (BjGRF19), qhov nruab nrab ntawm hydrophilicity. (BjGRF31) txog -0.45 (BjGRF22), tag nrho cov hydrophilic BjGRF proteins muaj qhov tsis zoo GRAVY qhov tseem ceeb, uas tej zaum yuav yog vim tsis muaj hydrophobicity los ntawm cov residues. Subcellular localization kwv yees pom tau tias 31 BjGRF encoded proteins tuaj yeem nyob hauv lub nucleus, BjGRF04 tuaj yeem nyob hauv peroxisomes, BjGRF25 tuaj yeem nyob hauv cytoplasm, thiab BjGRF28 tuaj yeem nyob hauv thaj chaw hauv chloroplasts (Table 1), qhia tias lub luag haujlwm tseem ceeb hauv zos thiab BjGRF. ib tug transcription factor.
Phylogenetic tsom xam ntawm GRF cov tsev neeg hauv ntau hom tuaj yeem pab tshawb nrhiav cov haujlwm ntawm noob. Yog li ntawd, tag nrho-ntev amino acid sequences ntawm 35 rapeseed, 16 turnip, 12 mov, 10 millet thiab 9 Arabidopsis GRFs tau downloaded thiab ib tsob ntoo phylogenetic tau tsim raws li 34 txheeb xyuas cov noob BjGRF (Fig. 1). Peb subfamilies muaj cov tswv cuab sib txawv; 116 GRF TFs tau muab faib ua peb pawg neeg sib txawv (pab pawg A ~ C), muaj 59 (50.86%), 34 (29.31%) thiab 23 (19.83)% ntawm GRFs, feem. Ntawm lawv, 34 BjGRF tsev neeg tau tawg ri niab nyob rau hauv 3 subfamilies: 13 cov tswv cuab hauv pawg A (38.24%), 12 cov tswv cuab hauv pawg B (35.29%) thiab 9 cov tswv cuab hauv pawg C (26.47%). Nyob rau hauv tus txheej txheem ntawm mustard polyploidization, tus naj npawb ntawm BjGRFs genes nyob rau hauv txawv subfamilies txawv, thiab gene amplification thiab poob yuav tshwm sim. Nws yog ib nqi sau cia hais tias tsis muaj kev faib ntawm mov thiab millet GRFs nyob rau hauv pawg C, thaum muaj 2 mov GRFs thiab 1 millet GRF nyob rau hauv pawg B, thiab feem ntau ntawm cov nplej thiab millet GRFs yog pab pawg nyob rau hauv ib ceg, qhia tau hais tias BjGRFs yog ze ze rau dicots. Ntawm lawv, qhov kev tshawb fawb tob tshaj plaws ntawm GRF muaj nuj nqi hauv Arabidopsis thaliana muab lub hauv paus rau kev kawm ua haujlwm ntawm BjGRFs.
Phylogenetic ntoo ntawm mustard suav nrog Brassica napus, Brassica napus, mov, millet thiab cov tswv cuab ntawm tsev neeg Arabidopsis thaliana GRF.
Kev tshuaj xyuas cov noob caj noob ces nyob rau hauv mustard GRF tsev neeg. Cov kab grey nyob rau hauv keeb kwm yav dhau los sawv cev rau ib qho kev sib koom ua ke nyob rau hauv mustard genome, cov kab liab sawv cev rau ib khub ntawm segmented rov ua dua ntawm BjGRF noob;
BjGRF gene qhia nyob rau hauv drought kev nyuaj siab ntawm plaub nplooj ntoos theem. qRT-PCR cov ntaub ntawv tau qhia hauv Cov Lus Ntxiv S5. Qhov sib txawv tseem ceeb hauv cov ntaub ntawv yog qhia los ntawm cov tsiaj ntawv me.
Raws li kev nyab xeeb thoob ntiaj teb hloov pauv mus ntxiv, kawm txog yuav ua li cas cov qoob loo tiv thaiv kev ntxhov siab vim kev ntxhov siab thiab kev txhim kho lawv cov txheej txheem kev zam tau dhau los ua qhov kev tshawb fawb kub 18. Tom qab drought, cov qauv morphological, noob qhia thiab cov txheej txheem metabolic ntawm cov nroj tsuag yuav hloov, uas yuav ua rau kom cov photosynthesis thiab metabolic cuam tshuam, cuam tshuam cov qoob loo thiab zoo ntawm cov qoob loo19,20,21. Thaum cov nroj tsuag hnov mob qhuav, lawv tsim cov tub txib thib ob xws li Ca2+ thiab phosphatidylinositol, ua rau kom cov calcium ion concentration ntawm intracellular thiab qhib kev tswj hwm kev sib txuas ntawm cov protein phosphorylation pathway22,23. Lub hom phiaj kawg ntawm cov protein yog koom ncaj qha rau hauv kev tiv thaiv cellular los yog tswj cov kev qhia ntawm cov noob muaj kev ntxhov siab los ntawm TFs, txhim kho cov nroj tsuag kam rau kev ntxhov siab 24,25. Yog li, TFs ua lub luag haujlwm tseem ceeb hauv kev teb rau kev ntxhov siab rau kev ntxhov siab. Raws li qhov sib lawv liag thiab DNA khi cov khoom ntawm drought stress responsive TFs, TFs tuaj yeem muab faib ua cov tsev neeg sib txawv, xws li GRF, ERF, MYB, WRKY thiab lwm tsev neeg26.
GRF gene tsev neeg yog ib hom nroj tsuag tshwj xeeb TF uas ua lub luag haujlwm tseem ceeb hauv ntau yam xws li kev loj hlob, kev loj hlob, kev hloov pauv thiab cov lus teb cog qoob loo. Txij li thaum thawj GRF noob tau txheeb xyuas hauv O. sativa28, ntau thiab ntau GRF cov noob tau txheeb pom nyob rau hauv ntau hom thiab pom tias cuam tshuam rau cov nroj tsuag kev loj hlob, kev loj hlob thiab kev ntxhov siab teb8, 29, 30,31,32. Nrog rau kev tshaj tawm ntawm Brassica juncea genome ib ntus, kev txheeb xyuas ntawm BjGRF gene tsev neeg tau ua tau33. Nyob rau hauv txoj kev tshawb no, 34 BjGRF noob tau txheeb xyuas nyob rau hauv tag nrho mustard genome thiab npe hu ua BjGRF01-BjGRF34 raws li lawv txoj haujlwm chromosomal. Tag nrho cov ntawm lawv muaj kev txuag QLQ thiab WRC cov thawj. Kev tshuaj xyuas ntawm cov khoom siv lub cev tau pom tias qhov sib txawv ntawm cov amino acid cov lej thiab qhov hnyav molecular ntawm BjGRF cov proteins (tshwj tsis yog BjGRF28) tsis tseem ceeb, qhia tau tias BjGRF tsev neeg yuav muaj cov haujlwm zoo sib xws. Gene qauv tsom xam pom tias 64.7% ntawm BjGRF cov noob muaj 4 exons, qhia tias BjGRF gene qauv tau txais txiaj ntsig zoo hauv evolution, tab sis tus naj npawb ntawm exons hauv BjGRF10, BjGRF16, BjGRP28 thiab BjGRF29 noob loj dua. Cov kev tshawb fawb tau pom tias qhov sib ntxiv lossis tshem tawm ntawm exons lossis introns tuaj yeem ua rau muaj qhov sib txawv ntawm cov qauv noob thiab kev ua haujlwm, yog li tsim cov noob tshiab34,35,36. Yog li ntawd, peb xav tias lub intron ntawm BjGRF tau ploj thaum lub sij hawm evolution, uas yuav ua rau muaj kev hloov pauv hauv cov noob muaj nuj nqi. Raws li cov kev tshawb fawb uas twb muaj lawm, peb kuj pom tau hais tias tus naj npawb ntawm introns tau txuam nrog gene qhia. Thaum tus naj npawb ntawm introns hauv cov noob loj, cov noob tuaj yeem teb sai sai rau ntau yam tsis zoo.
Gene duplication yog ib qho tseem ceeb hauv genomic thiab genetic evolution37. Cov kev tshawb fawb cuam tshuam tau pom tias cov noob duplication tsis tsuas yog nce tus naj npawb ntawm GRF cov noob, tab sis kuj yog ib txoj hauv kev tsim cov noob tshiab los pab cov nroj tsuag hloov mus rau ntau yam tsis zoo ib puag ncig38. Tag nrho ntawm 48 qhov sib npaug ntawm cov noob caj noob ces tau pom nyob rau hauv txoj kev tshawb no, tag nrho cov uas yog segmental duplications, qhia tau hais tias segmental duplications yog lub luag haujlwm tseem ceeb rau kev nce cov noob hauv tsev neeg no. Nws tau raug tshaj tawm nyob rau hauv cov ntaub ntawv hais tias segmental duplication muaj peev xwm txhawb nqa lub amplification ntawm GRF noob tsev neeg nyob rau hauv Arabidopsis thiab strawberry, thiab tsis muaj tandem duplication ntawm no gene tsev neeg tau pom nyob rau hauv ib yam ntawm cov hom27,39. Cov txiaj ntsig ntawm txoj kev tshawb fawb no tau ua raws li cov kev tshawb fawb uas twb muaj lawm ntawm Arabidopsis thaliana thiab cov tsev neeg strawberry, qhia tias tsev neeg GRF tuaj yeem ua rau muaj cov noob thiab tsim cov noob tshiab los ntawm cov segmental duplication hauv cov nroj tsuag sib txawv.
Nyob rau hauv txoj kev tshawb no, tag nrho ntawm 34 BjGRF noob tau txheeb xyuas nyob rau hauv mustard, uas tau muab faib ua 3 subfamilies. Cov noob no tau pom zoo li kev khaws cia motifs thiab cov qauv noob. Collinearity tsom xam qhia 48 khub ntawm ntu duplications hauv mustard. BjGRF txhawb nqa cheeb tsam muaj cov cis-ua yeeb yam cuam tshuam nrog lub teeb teb, cov lus teb hormonal, ib puag ncig kev ntxhov siab, thiab kev loj hlob thiab kev loj hlob. Qhov kev qhia ntawm 34 BjGRF noob tau kuaj pom nyob rau theem ntawm cov noob txiv ntoo (cov hauv paus, stems, nplooj), thiab cov qauv qhia ntawm 10 BjGRF noob nyob rau hauv drought tej yam kev mob. Nws tau pom tias cov qauv qhia ntawm BjGRF cov noob nyob rau hauv kev ntxhov siab ntawm kev ntxhov siab yog zoo sib xws thiab tej zaum yuav zoo sib xws. kev koom tes hauv drought Forcing regulation. BjGRF03 thiab BjGRF32 cov noob tuaj yeem ua lub luag haujlwm zoo hauv kev ntxhov siab, thaum BjGRF06 thiab BjGRF23 ua lub luag haujlwm hauv kev ntxhov siab ntxhov siab li miR396 lub hom phiaj noob. Zuag qhia tag nrho, peb txoj kev tshawb fawb muab lub hauv paus lom neeg rau kev tshawb pom yav tom ntej ntawm BjGRF gene muaj nuj nqi hauv cov nroj tsuag Brassicaceae.
Cov noob mustard siv hauv qhov kev sim no tau muab los ntawm Guizhou Oil Seed Research Institute, Guizhou Academy of Agricultural Sciences. Xaiv tag nrho cov noob thiab cog rau hauv av (substrate: av = 3: 1), thiab sau cov hauv paus hniav, stems thiab nplooj tom qab plaub-nplooj theem. Cov nroj tsuag tau kho nrog 20% PEG 6000 los simulate drought, thiab nplooj tau sau tom qab 0, 3, 6, 12 thiab 24 teev. Tag nrho cov qauv cog tau khov tam sim ntawd hauv cov kua nitrogen thiab tom qab ntawd muab cia rau hauv -80 ° C lub freezer rau kev sim tom ntej.
Tag nrho cov ntaub ntawv tau los yog tshuaj xyuas thaum lub sij hawm txoj kev tshawb no muaj nyob rau hauv tsab xov xwm luam tawm thiab ntxiv cov ntaub ntawv ntxiv.
Post lub sij hawm: Jan-22-2025