Kufalikira kwa mvula m'nyengo zosiyanasiyana ku Guizhou Province sikofanana, ndipo mvula imagwa kwambiri m'nyengo ya masika ndi chilimwe, koma mbande za rapeseed zimavutika ndi chilala nthawi yophukira ndi yozizira, zomwe zimakhudza kwambiri zokolola. Mustard ndi mbewu yapadera yamafuta yomwe imalimidwa makamaka ku Guizhou Province. Imapirira chilala kwambiri ndipo imatha kulimidwa m'mapiri. Ndi gwero lolemera la majini olimbana ndi chilala. Kupezeka kwa majini olimbana ndi chilala ndikofunikira kwambiri pakukweza mitundu ya mpiru. komanso kupanga zatsopano muzinthu za germplasm. Banja la GRF limagwira ntchito yofunika kwambiri pakukula ndi chitukuko cha zomera komanso kuthana ndi chilala. Pakadali pano, majini a GRF apezeka mu Arabidopsis 2, mpunga (Oryza sativa) 12, rapeseed 13, thonje (Gossypium hirsutum) 14, tirigu (Triticum). aestivum)15, pearl millet (Setaria italica)16 ndi Brassica17, koma palibe malipoti a majini a GRF omwe apezeka mu mpiru. Mu kafukufukuyu, majini a banja la mpiru a GRF adazindikirika pamlingo wonse wa majini ndipo mawonekedwe awo akuthupi ndi a mankhwala, ubale wawo wosinthika, kufanana kwawo, mawonekedwe osungidwa, kapangidwe ka majini, kubwerezabwereza kwa majini, zinthu za cis ndi gawo la mbande (gawo la masamba anayi) zidasanthulidwa. Mawonekedwe a mawonekedwe omwe ali pansi pa chilala adasanthulidwa mokwanira kuti apereke maziko asayansi a maphunziro ena okhudza momwe majini a BjGRF angagwiritsidwire ntchito poyankha chilala komanso kupereka majini oyenera kuswana mpiru wopirira chilala.
Majini makumi atatu mphambu anayi a BjGRF adapezeka mu genome ya Brassica juncea pogwiritsa ntchito kufufuza kawiri kwa HMMER, komwe konse kuli madera a QLQ ndi WRC. Ma CDS sequences a majini a BjGRF odziwika aperekedwa mu Supplementary Table S1. BjGRF01–BjGRF34 adatchulidwa kutengera malo awo pa chromosome. Kapangidwe ka physicochemical ka banjali kakusonyeza kuti kutalika kwa amino acid kumasinthasintha kwambiri, kuyambira 261 aa (BjGRF19) mpaka 905 aa (BjGRF28). Malo oyeretsera a BjGRF amachokera pa 6.19 (BjGRF02) mpaka 9.35 (BjGRF03) ndi avareji ya 8.33, ndipo 88.24% ya BjGRF ndi puloteni yoyambira. Kulemera kwa molekyulu ya BjGRF komwe kunanenedweratu ndi kuyambira 29.82 kDa (BjGRF19) mpaka 102.90 kDa (BjGRF28); index yosakhazikika ya mapuloteni a BjGRF imayambira pa 51.13 (BjGRF08) mpaka 78.24 (BjGRF19), onse ndi opitilira 40, zomwe zikusonyeza kuti index ya fatty acid imayambira pa 43.65 (BjGRF01) mpaka 78.78 (BjGRF22), avareji ya hydrophilicity (GRAVY) imayambira pa -1.07 (BjGRF31) mpaka -0.45 (BjGRF22), mapuloteni onse a BjGRF okonda hydrophilic ali ndi GRAVY yoyipa, zomwe zitha kukhala chifukwa cha kusowa kwa hydrophobicity komwe kumachitika chifukwa cha zotsalira. Kuneneratu kwa malo a subcellular kunawonetsa kuti mapuloteni 31 a BjGRF omwe ali ndi ma codec akhoza kupezeka mu nyukiliyasi, BjGRF04 ikhoza kupezeka mu peroxisomes, BjGRF25 ikhoza kupezeka mu cytoplasm, ndipo BjGRF28 ikhoza kupezeka mu ma chloroplasts (Table 1), zomwe zikusonyeza kuti ma BjGRF akhoza kupezeka mu nyukiliyasi ndikuchita gawo lofunikira pakulamulira ngati chinthu cholembera.
Kusanthula kwa phylogenetic kwa mabanja a GRF m'mitundu yosiyanasiyana kungathandize kuphunzira ntchito za majini. Chifukwa chake, ma amino acid athunthu a mbewu 35 za rapeseed, turnip 16, mpunga 12, mapira 10 ndi 9 za Arabidopsis GRF adatsitsidwa ndipo mtengo wa phylogenetic unapangidwa kutengera majini 34 a BjGRF omwe adadziwika (Chithunzi 1). Mabanja atatuwa ali ndi ziwerengero zosiyana za mamembala; Ma GRF TF 116 amagawidwa m'mabanja atatu osiyana (magulu A~C), okhala ndi 59 (50.86%), 34 (29.31%) ndi 23 (19.83)% ya ma GRF, motsatana. Pakati pawo, mamembala 34 a banja la BjGRF ali m'mabanja atatu ang'onoang'ono: mamembala 13 m'gulu A (38.24%), mamembala 12 m'gulu B (35.29%) ndi mamembala 9 m'gulu C (26.47%). Mu ndondomeko ya mustard polyploidization, chiwerengero cha majini a BjGRF m'mabanja osiyanasiyana ndi chosiyana, ndipo kukulitsa ndi kutayika kwa majini kungakhale kuti kwachitika. Ndikofunikira kudziwa kuti palibe kufalikira kwa mpunga ndi mapira a GRF mu gulu C, pomwe pali ma GRF awiri a mpunga ndi mapira a GRF mu gulu B, ndipo ma GRF ambiri a mpunga ndi mapira ali m'gulu limodzi, zomwe zikusonyeza kuti ma BjGRF ndi ofanana kwambiri ndi ma dicots. Pakati pawo, maphunziro ozama kwambiri pa ntchito ya GRF mu Arabidopsis thaliana amapereka maziko a maphunziro ogwira ntchito a ma BjGRF.
Mtengo wa mpiru wa phylogenetic kuphatikizapo Brassica napus, Brassica napus, mpunga, mapira ndi mamembala a banja la Arabidopsis thaliana GRF.
Kusanthula majini obwerezabwereza m'banja la mpiru wa GRF. Mzere wakuda kumbuyo ukuyimira chipika chogwirizana mu genome ya mpiru, mzere wofiira ukuyimira kubwereza kawiri kwa jini ya BjGRF;
Kuwonekera kwa majini a BjGRF pansi pa chilala pa gawo lachinayi la tsamba. Deta ya qRT-PCR ikuwonetsedwa mu Supplementary Table S5. Kusiyana kwakukulu kwa deta kukuwonetsedwa ndi zilembo zazing'ono.
Pamene nyengo yapadziko lonse ikupitirira kusintha, kuphunzira momwe mbewu zimathanirana ndi vuto la chilala ndikuwongolera njira zawo zolekerera kwakhala nkhani yofunika kwambiri yofufuza18. Pambuyo pa chilala, kapangidwe ka mawonekedwe, mawonekedwe a majini ndi kagayidwe kachakudya ka zomera zidzasintha, zomwe zingayambitse kutha kwa photosynthesis ndi kusokonezeka kwa kagayidwe kachakudya, zomwe zimakhudza zokolola ndi ubwino wa mbewu19,20,21. Zomera zikamamva zizindikiro za chilala, zimapanga amithenga achiwiri monga Ca2+ ndi phosphatidylinositol, zimawonjezera kuchuluka kwa calcium ion mkati mwa maselo ndikuyambitsa netiweki yolamulira ya njira ya phosphorylation ya mapuloteni22,23. Puloteni yomaliza yomwe cholinga chake ndi kuteteza maselo kapena imayang'anira kuwonetsedwa kwa majini okhudzana ndi kupsinjika kudzera mu TFs, kukulitsa kupirira kwa zomera ku kupsinjika24,25. Chifukwa chake, TFs zimakhala ndi gawo lofunikira poyankha ku kupsinjika kwa chilala. Malinga ndi momwe DNA imagwirizanirana ndi ma TFs omwe amayankha kupsinjika kwa chilala, TFs zimatha kugawidwa m'mabanja osiyanasiyana, monga GRF, ERF, MYB, WRKY ndi mabanja ena26.
Banja la majini a GRF ndi mtundu wa TF yochokera ku zomera yomwe imagwira ntchito zofunika kwambiri m'mbali zosiyanasiyana monga kukula, chitukuko, kufalitsa zizindikiro ndi mayankho a chitetezo cha zomera27. Kuyambira pamene jini loyamba la GRF linapezeka mu O. sativa28, majini ambiri a GRF apezeka m'mitundu yambiri ndipo awonetsedwa kuti amakhudza kukula kwa zomera, chitukuko ndi kuyankha kupsinjika8, 29, 30,31,32. Ndi kufalitsidwa kwa Brassica juncea genome sequence, kuzindikira banja la majini a BjGRF kunakhala kotheka33. Mu kafukufukuyu, majini 34 a BjGRF adapezeka mu genome yonse ya mpiru ndipo adatchedwa BjGRF01–BjGRF34 kutengera malo awo a chromosome. Onsewa ali ndi madera osungidwa bwino a QLQ ndi WRC. Kusanthula kwa makhalidwe a physicochemical kunawonetsa kuti kusiyana kwa manambala a amino acid ndi kulemera kwa mamolekyu a mapuloteni a BjGRF (kupatula BjGRF28) sikunali kofunikira, zomwe zikusonyeza kuti mamembala a banja la BjGRF akhoza kukhala ndi ntchito zofanana. Kusanthula kapangidwe ka majini kunasonyeza kuti 64.7% ya majini a BjGRF anali ndi ma exon anayi, zomwe zikusonyeza kuti kapangidwe ka majini a BjGRF kamasungidwa pang'ono pakusintha kwa zinthu, koma chiwerengero cha ma exon mu majini a BjGRF10, BjGRF16, BjGRP28 ndi BjGRF29 ndi chachikulu. Kafukufuku wasonyeza kuti kuwonjezera kapena kuchotsa ma exon kapena ma intron kungayambitse kusiyana kwa kapangidwe ka majini ndi ntchito zake, motero kupanga majini atsopano 34,35,36. Chifukwa chake, tikuganiza kuti intron ya BjGRF inatayika panthawi ya kusintha kwa zinthu, zomwe zingayambitse kusintha kwa ntchito ya majini. Mogwirizana ndi kafukufuku omwe alipo, tapezanso kuti chiwerengero cha ma intron chinali chogwirizana ndi kufotokozera kwa majini. Pamene chiwerengero cha ma intron mu jini chili chachikulu, jini imatha kuyankha mwachangu pazinthu zosiyanasiyana zoyipa.
Kubwerezabwereza majini ndi chinthu chachikulu pakusintha kwa majini ndi majini37. Kafukufuku wofanana wasonyeza kuti kubwerezabwereza majini sikuti kumawonjezera chiwerengero cha majini a GRF okha, komanso kumagwira ntchito ngati njira yopangira majini atsopano kuti athandize zomera kuzolowera nyengo zosiyanasiyana zoyipa38. Ma gene awiriawiri okwana 48 obwerezabwereza adapezeka mu kafukufukuyu, onse omwe anali kubwerezabwereza kwa magawo, zomwe zikusonyeza kuti kubwerezabwereza kwa magawo ndiye njira yayikulu yowonjezera chiwerengero cha majini m'banjali. Zanenedwa m'mabuku kuti kubwerezabwereza kwa magawo kungalimbikitse bwino kukulitsa kwa mamembala a majini a GRF mu Arabidopsis ndi sitiroberi, ndipo palibe kubwerezabwereza kwa banja la majini awa komwe kudapezeka mu mtundu uliwonse27,39. Zotsatira za kafukufukuyu zikugwirizana ndi maphunziro omwe alipo pa mabanja a Arabidopsis thaliana ndi sitiroberi, zomwe zikusonyeza kuti banja la GRF likhoza kuwonjezera chiwerengero cha majini ndikupanga majini atsopano kudzera mu kubwerezabwereza kwa magawo m'zomera zosiyanasiyana.
Mu kafukufukuyu, majini 34 a BjGRF adapezeka mu mpiru, omwe adagawidwa m'mabanja atatu ang'onoang'ono. Majini awa adawonetsa mapangidwe ofanana osungidwa ndi kapangidwe ka majini. Kusanthula kwa Collinearity kudavumbulutsa magawo 48 a kubwerezabwereza kwa magawo mu mpiru. Dera lothandizira la BjGRF lili ndi zinthu zomwe zimagwira ntchito ya cis zomwe zimagwirizanitsidwa ndi kuyankhidwa kwa kuwala, kuyankhidwa kwa mahomoni, kuyankhidwa kwa kupsinjika kwa chilengedwe, ndi kukula ndi chitukuko. Kuwonetsedwa kwa majini 34 a BjGRF kudapezeka pagawo la mmera wa mpiru (mizu, tsinde, masamba), ndi mawonekedwe a majini 10 a BjGRF pansi pa chilala. Zapezeka kuti mawonekedwe a majini a BjGRF pansi pa chilala anali ofanana ndipo akhoza kukhala ofanana. kutenga nawo mbali mu chilala. Kukakamiza malamulo. Majini a BjGRF03 ndi BjGRF32 akhoza kukhala ndi maudindo abwino olamulira pakupsinjika kwa chilala, pomwe majini a BjGRF06 ndi BjGRF23 amasewera maudindo pakupsinjika kwa chilala ngati majini omwe amafunidwa ndi miR396. Ponseponse, kafukufuku wathu amapereka maziko achilengedwe opezera ntchito ya majini a BjGRF mtsogolo mu zomera za Brassicaceae.
Mbewu za mpiru zomwe zinagwiritsidwa ntchito mu kuyesera kumeneku zinaperekedwa ndi Guizhou Oil Seed Research Institute, Guizhou Academy of Agricultural Sciences. Sankhani mbewu zonse ndikuzibzala m'nthaka (nthaka yonyowa = 3:1), ndikusonkhanitsa mizu, tsinde ndi masamba pambuyo pa gawo la masamba anayi. Zomerazo zinapatsidwa mankhwala a 20% PEG 6000 kuti ziyerekezere chilala, ndipo masambawo anasonkhanitsidwa patatha maola 0, 3, 6, 12 ndi 24. Zitsanzo zonse za zomera zinazizidwa nthawi yomweyo mu nayitrogeni yamadzimadzi kenako nkusungidwa mufiriji ya -80°C kuti muyesedwenso.
Deta yonse yomwe yapezedwa kapena kufufuzidwa panthawi ya kafukufukuyu ikuphatikizidwa mu nkhani yofalitsidwa komanso mafayilo owonjezera azidziwitso.
Nthawi yotumizira: Januwale-22-2025