Ikwirakwizwa ry’ibihe mu Ntara ya Guizhou ntiriringaniye, hamwe n’imvura nyinshi mu mpeshyi no mu cyi, ariko ingemwe zafashwe ku ngufu zishobora kwibasirwa n’amapfa mu gihe cyizuba n'itumba, bigira ingaruka zikomeye ku musaruro. Sinapi ni igihingwa kidasanzwe cyamavuta gihingwa cyane mu Ntara ya Guizhou. Ifite kwihanganira amapfa kandi irashobora guhingwa mumisozi. Numutungo ukungahaye wa gen irwanya amapfa. Ivumburwa rya gen irwanya amapfa ningirakamaro cyane mugutezimbere ubwoko bwa sinapi. no guhanga udushya muri germplasm. Umuryango GRF ugira uruhare runini mu mikurire no gutera imbere no guhangana n’amapfa. Kugeza ubu, GRF yabonetse muri Arabidopsis 2, umuceri (Oryza sativa) 12, gufata kungufu 13, ipamba (Gossypium hirsutum) 14, ingano (Triticum). aestivum) 15, isaro (Setaria italica) 16 na Brassica17, ariko nta makuru yerekana genes za GRF zagaragaye muri sinapi. Muri ubu bushakashatsi, isesengura ry’imiryango ya GRF ya sinapi ryagaragaye ku rwego rwa genome yose kandi biranga umubiri na shimi biranga umubiri, isano y'ubwihindurize, homologiya, imiterere yabitswe, imiterere ya gene, kwigana gene, cis-element hamwe nicyiciro cyo gutera (icyiciro cy'amababi ane). Uburyo bwo kwerekana ibibazo by’amapfa bwasesenguwe ku buryo bwuzuye kugira ngo butange ubumenyi bwa siyansi kugira ngo hakorwe ubundi bushakashatsi ku mikorere ishobora kuba ingirabuzimafatizo za BjGRF mu guhangana n’amapfa no gutanga genes z’abakandida zo korora sinapi yihanganira amapfa.
Imirongo 34 ya BjGRF yamenyekanye muri genome ya Brassica juncea ikoresheje ubushakashatsi bubiri bwa HMMER, yose irimo QLQ na WRC. Urutonde rwa CDS rwerekana genes za BjGRF zerekanwe kumeza yinyongera S1. BjGRF01 - BjGRF34 yitwa amazina ukurikije aho bari kuri chromosome. Imiterere ya fiziki ya chimique yuyu muryango yerekana ko uburebure bwa aside amine butandukanye cyane, kuva kuri 261 aa (BjGRF19) kugeza 905 aa (BjGRF28). Ingingo ya isoelectric ya BjGRF iri hagati ya 6.19 (BjGRF02) kugeza 9.35 (BjGRF03) ikigereranyo cya 8.33, naho 88.24% ya BjGRF ni proteine yibanze. Ibipimo byerekana uburemere bwa BjGRF ni kuva 29.82 kDa (BjGRF19) kugeza kuri 102.90 kDa (BjGRF28); igipimo kidahungabana cya poroteyine za BjGRF kiri hagati ya 51.13 (BjGRF08) kugeza kuri 78.24 (BjGRF19), byose birarenze 40, byerekana ko igipimo cya aside irike kiva kuri 43.65 (BjGRF01) kugeza kuri 78.78 (BjGRF22), ugereranije na hydrophilicity (GRAVY07) . Ihanurwa rya subcellular ryerekanaga ko poroteyine 31 za BjGRF zishobora gushyirwa muri nucleus, BjGRF04 ishobora kuba muri peroxisomes, BjGRF25 ishobora kuba muri cytoplazme, naho BjGRF28 ikaba ishobora kuba muri chloroplasts (Imbonerahamwe 1), byerekana ko BjGRFs ishobora kugira uruhare runini muri nucleus.
Isesengura rya phylogeneque yimiryango ya GRF mubwoko butandukanye irashobora gufasha kwiga imikorere ya gene. Kubera iyo mpamvu, harakuweho uburebure bwa aminide acide ya amine 35 ya kungufu 35, shitingi 16, umuceri 12, umuceri 10 na 9 Arabidopsis GRFs hanyuma hubakwa igiti cyitwa phylogeneque gishingiye kuri gen 34 za BjGRF zamenyekanye (Ishusho 1). Imiryango itatu igizwe numubare utandukanye wabanyamuryango; 116 GRF TF igabanijwemo imiryango itatu itandukanye (amatsinda A ~ C), irimo 59 (50.86%), 34 (29.31%) na 23 (19.83)% ya GRFs. Muri bo, 34 bo mu muryango wa BjGRF batatanye mu miryango 3: Abanyamuryango 13 mu itsinda A (38.24%), abanyamuryango 12 mu itsinda B (35.29%) n’abanyamuryango 9 mu itsinda C (26.47%). Mubikorwa bya sinapi polyploidisation, umubare wa gen ya BjGRFs mumiryango itandukanye iratandukanye, kandi amplification no gutakaza gene bishobora kuba byarabaye. Twabibutsa ko nta gukwirakwiza umuceri na umuceri GRFs mu itsinda C, mu gihe hari GRFs 2 z'umuceri na GRF 1 z'umuceri mu itsinda B, kandi ibyinshi mu muceri n'umuceri GRFs bishyizwe mu ishami rimwe, byerekana ko BjGRF ifitanye isano rya hafi na dicot. Muri byo, ubushakashatsi bwimbitse ku mikorere ya GRF muri Arabidopsis thaliana butanga urufatiro rwo kwiga imikorere ya BjGRFs.
Igiti cya phylogeneque cya sinapi harimo Brassica napus, Brassica napus, umuceri, umuceri hamwe nabagize umuryango wa Arabidopsis thaliana GRF.
Isesengura rya genes zisubiramo mumuryango wa sinapi GRF. Umurongo wijimye winyuma ugereranya guhuza muri genome ya sinapi, umurongo utukura ugereranya ibice bibiri byisubiramo bya gen ya BjGRF;
Imvugo ya BjGRF mugihe cy'amapfa kurwego rwa kane rwibabi. qRT-PCR amakuru yerekanwe mumeza yinyongera S5. Itandukaniro rikomeye mumibare ryerekanwa ninyuguti nto.
Mugihe ikirere cyisi gikomeje guhinduka, kwiga uburyo ibihingwa bihangayikishwa n’amapfa no kunoza uburyo bwo kwihanganira byahindutse ingingo y’ubushakashatsi18. Nyuma y’amapfa, imiterere ya morfologiya, imiterere ya gene hamwe nuburyo bwo guhinduranya ibimera bizahinduka, ibyo bikaba bishobora gutuma fotosintezeza ihagarara ndetse n’imivurungano ya metabolike, bikagira ingaruka ku musaruro nubwiza bwibihingwa 19,20,21. Iyo ibimera byunvise ibimenyetso byamapfa, bitanga ubutumwa bwa kabiri nka Ca2 + na phosphatidylinositol, byongera intungamubiri za calcium ion hamwe no gukora imiyoboro igenga inzira ya protein fosifora 22,23. Intego ya nyuma ya poroteyine igira uruhare rutaziguye mu kurinda selile cyangwa igenga imiterere ya genes zifitanye isano na TFs, bigatuma kwihanganira ibimera bigabanuka 24,25. Rero, TF igira uruhare runini mugukemura ibibazo byamapfa. Ukurikije urutonde hamwe na ADN ihuza imiterere y amapfa yatewe n amapfa, TF irashobora kugabanywa mumiryango itandukanye, nka GRF, ERF, MYB, WRKY nindi miryango26.
Umuryango wa GRF ni ubwoko bwibimera byihariye bya TF bigira uruhare runini mubice bitandukanye nko gukura, iterambere, guhererekanya ibimenyetso no gusubiza ibihingwa27. Kuva gene ya mbere ya GRF yamenyekana muri O. sativa28, genes nyinshi ninshi za GRF zagaragaye mumoko menshi kandi zerekanwa ko zigira ingaruka kumikurire yibimera, iterambere no guhangayika8, 29, 30,31,32. Hamwe nogukurikirana genome ya Brassica juncea, kumenyekanisha umuryango wa BjGRF byashobokaga33. Muri ubu bushakashatsi, gen 34 za BjGRF zagaragaye muri genome yose ya sinapi kandi yitwa BjGRF01 - BjGRF34 ukurikije imyanya yabo ya chromosomal. Byose birimo QLQ yabitswe cyane na WRC. Isesengura ryimiterere ya fiziki ya chimique ryerekanye ko itandukaniro ryimibare ya aside amine nuburemere bwa molekuline za poroteyine za BjGRF (usibye BjGRF28) bitari ngombwa, byerekana ko abagize umuryango wa BjGRF bashobora kuba bafite imirimo isa. Isesengura ryimiterere ya gen ryerekanye ko 64.7% ya gen ya BjGRF yarimo exons 4, byerekana ko imiterere ya gene ya BjGRF yabitswe cyane mubwihindurize, ariko umubare wa exons muri BjGRF10, BjGRF16, BjGRP28 na BjGRF29 ni munini. Ubushakashatsi bwerekanye ko kongeramo cyangwa gusiba exons cyangwa intron bishobora gutera itandukaniro mumiterere ya gene n'imikorere, bityo bigakora genes nshya 34,35,36. Kubwibyo, turakeka ko intron ya BjGRF yatakaye mugihe cyubwihindurize, ishobora gutera impinduka mumikorere ya gene. Dukurikije ubushakashatsi buriho, twasanze kandi umubare winjirana wajyanye no kwerekana gene. Iyo umubare winjira muri gene ari munini, gene irashobora guhita isubiza ibintu bitandukanye bitameze neza.
Kwigana kw'ibinyabuzima ni ikintu gikomeye mu bwihindurize bwa genoside na genetike37. Ubushakashatsi bujyanye na bwo bwerekanye ko kwigana gene bitongera umubare wa GRF gusa, ahubwo binakora nk'uburyo bwo kubyara ingirabuzimafatizo nshya zifasha ibimera guhuza n'ibidukikije bitandukanye 38. Muri ubu bushakashatsi habonetse 48 ebyiri zibiri za gene ebyiri zabigenewe, zose zikaba ari duplication segment, byerekana ko kwigana ibice aribwo buryo bukomeye bwo kongera umubare wa gen muri uyu muryango. Mu bitabo byavuzwe ko kwigana ibice bishobora guteza imbere kongera ingufu mu muryango wa GRF gene muri Arabidopsis na strawberry, kandi nta kwigana tandem y’uyu muryango wabonetse muri bumwe mu bwoko 27,39. Ibyavuye muri ubu bushakashatsi bihuye n’ubushakashatsi buriho ku miryango ya Arabidopsis thaliana na strawberry, byerekana ko umuryango GRF ushobora kongera umubare wa gen kandi ukabyara genes nshya binyuze mu kwigana ibice mu bimera bitandukanye.
Muri ubu bushakashatsi, hagaragaye genes 34 za BjGRF muri sinapi, zigabanyijemo imiryango 3. Izi genes zerekanaga motif zabitswe hamwe nuburyo bwa gene. Isesengura rya collinearity ryerekanye ibice 48 byo kwigana muri sinapi. Agace ka BjGRF gateza imbere karimo ibintu bifatika bifitanye isano no gusubiza urumuri, igisubizo cya hormone, guhangana n’ibidukikije, no gukura niterambere. Imvugo ya gen 34 ya BjGRF yagaragaye mugihe cyo gutera imbuto ya sinapi (imizi, uruti, amababi), hamwe nimiterere ya gen 10 ya BjGRF mugihe cyamapfa. Byagaragaye ko imvugo ya gen ya BjGRF ihangayikishijwe n amapfa isa kandi ishobora kuba imwe. uruhare mu ruzuba Guhatira amabwiriza. Imirasire ya BjGRF03 na BjGRF32 irashobora kugira uruhare runini muguhagarika amapfa, mugihe BjGRF06 na BjGRF23 bigira uruhare mukibazo cyamapfa nka miR396 gen. Muri rusange, ubushakashatsi bwacu butanga ishingiro ryibinyabuzima byo kuvumbura imikorere ya gene ya BjGRF mu bimera bya Brassicaceae.
Imbuto za sinapi zikoreshwa muri ubu bushakashatsi zatanzwe n’ikigo cy’ubushakashatsi cy’imbuto za Guizhou, Ishuri Rikuru ry’Ubuhinzi rya Guizhou. Hitamo imbuto zose hanyuma uzite mu butaka (substrate: ubutaka = 3: 1), hanyuma ukusanyirize imizi, ibiti n'amababi nyuma yicyiciro cyibabi bine. Ibimera byavuwe hamwe na 20% PEG 6000 kugirango bigereranye amapfa, kandi amababi yakusanyijwe nyuma yamasaha 0, 3, 6, 12 na 24. Ingero zose z’ibimera zahise zikonjeshwa muri azote yuzuye hanyuma zibikwa muri firigo -80 ° C kugirango ikizamini gikurikiraho.
Amakuru yose yabonetse cyangwa yasesenguwe muri ubu bushakashatsi ashyirwa mu ngingo yatangajwe hamwe namadosiye yinyongera.
Igihe cyo kohereza: Mutarama-22-2025