Kugawa kwanyengo kwa mvula m'chigawo cha Guizhou sikuli kofanana, kumakhala mvula yambiri mu kasupe ndi chilimwe, koma mbande za rapeseed zimatha kuvutika ndi chilala m'dzinja ndi m'nyengo yozizira, zomwe zimakhudza kwambiri zokolola. Mustard ndi mbewu yapadera yamafuta yomwe imabzalidwa makamaka m'chigawo cha Guizhou. Ali ndi chilala cholimba ndipo amatha kulimidwa m'madera amapiri. Ndilo chuma chambiri cha majini olimbana ndi chilala. Kupezeka kwa majini olimbana ndi chilala ndikofunikira kwambiri pakuwongolera mitundu ya mpiru. ndi zatsopano muzinthu za germplasm. Banja la GRF limagwira ntchito yofunika kwambiri pakukula kwa zomera ndi chitukuko komanso kuyankha ku mavuto a chilala. Pakali 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 amtundu wa GRF opezeka mu mustard. Mu phunziro ili, chibadwa cha GRF chamtundu wa mpiru chinadziwika pa mlingo wa genome-wide ndi mawonekedwe awo a thupi ndi mankhwala, maubwenzi osinthika, homology, zosungidwa zosungidwa, mapangidwe a jini, kubwereza kwa majini, cis-elements ndi siteji ya mmera (gawo la masamba anayi) adawunikidwa. Mafotokozedwe omwe akukumana ndi vuto la chilala adawunikidwa mozama kuti apereke maziko asayansi opitilira maphunziro owonjezera pakugwira ntchito kwa majini a BjGRF pothana ndi chilala komanso kupereka majini oti azitha kuswana mpiru wopirira chilala.
Majini makumi atatu ndi anayi a BjGRF adadziwika mu Brassica juncea genome pogwiritsa ntchito kusaka kuwiri kwa HMMER, komwe kuli madera a QLQ ndi WRC. Zotsatira za CDS za jini za BjGRF zodziwika zimaperekedwa mu Supplementary Table S1. BjGRF01–BjGRF34 amatchulidwa kutengera komwe ali pa chromosome. Ma physicochemical properties a banja ili amasonyeza kuti kutalika kwa amino acid kumakhala kosiyana kwambiri, kuyambira 261 aa (BjGRF19) mpaka 905 aa (BjGRF28). The isoelectric point of BjGRF ranges from 6.19 (BjGRF02) to 9.35 (BjGRF03) with avareji 8.33, ndi 88.24% ya BjGRF ndi zofunika mapuloteni. Zonenedweratu za kulemera kwa maselo a BjGRF zimachokera ku 29.82 kDa (BjGRF19) mpaka 102.90 kDa (BjGRF28); instability index of BjGRF protein ranges from 51.13 (BjGRF08) to 78.24 (BjGRF19), onse ndi aakulu kuposa 40, kusonyeza kuti fatty acid index ranges from 43.65 (BjGRF01) to 78.78 (BjGRF22), avareji 70 hydro.AVYphilicity (GGR) (BjGRF31) mpaka -0.45 (BjGRF22), mapuloteni onse a hydrophilic BjGRF ali ndi makhalidwe oipa a GRAVY, omwe angakhale chifukwa cha kusowa kwa hydrophobicity chifukwa cha zotsalira. Kuneneratu kwa malo amtundu wa subcellular kunawonetsa kuti mapuloteni osungidwa a 31 BjGRF amatha kupezeka mu nyukiliya, BjGRF04 ikhoza kukhazikitsidwa mu peroxisomes, BjGRF25 ikhoza kukhazikitsidwa mu cytoplasm, ndipo BjGRF28 ikhoza kukhazikitsidwa mu chloroplasts (Table 1), kusonyeza kuti BjGRF25 ikhoza kukhala gawo lofunikira la nucleus. cholembera.
Kusanthula kwa Phylogenetic kwa mabanja a GRF mumitundu yosiyanasiyana kungathandize kuphunzira ntchito za majini. Choncho, ma amino acid amtundu wa 35 a rapeseed, 16 mpiru, 12 mpunga, mapira 10 ndi 9 Arabidopsis GRFs adatsitsidwa ndipo mtengo wa phylogenetic unamangidwa potengera 34 jini ya BjGRF (mkuyu 1). Mabanja atatuwa ali ndi ziwerengero zosiyanasiyana za mamembala; 116 GRF TFs amagawidwa m'magulu atatu osiyana (magulu A~C), okhala ndi 59 (50.86%), 34 (29.31%) ndi 23 (19.83)% a GRFs, motsatana. Pakati pawo, 34 achibale a BjGRF amwazikana m'mabanja atatu: mamembala 13 a gulu A (38.24%), mamembala 12 a gulu B (35.29%) ndi 9 a gulu C (26.47%). M'kati mwa mpiru wa polyploidization, kuchuluka kwa ma jini a BjGRF m'mabanja osiyanasiyana ndi osiyana, ndipo kukulitsa kwa majini ndi kutayika kungakhalepo. Ndikoyenera kudziwa kuti palibe kugawidwa kwa ma GRF a mpunga ndi mapira mu gulu C, pamene pali 2 mpunga GRFs ndi 1 mapira GRF mu gulu B, ndipo ambiri a mpunga ndi mapira GRFs ali m'gulu limodzi nthambi, kusonyeza kuti BjGRFs zogwirizana kwambiri dicots. Pakati pawo, maphunziro ozama kwambiri pa ntchito ya GRF mu Arabidopsis thaliana amapereka maziko a maphunziro ogwira ntchito a BjGRF.
Mtengo wa Phylogenetic wa mpiru kuphatikiza Brassica napus, Brassica napus, mpunga, mapira ndi mamembala a banja la Arabidopsis thaliana GRF.
Kusanthula kwa majini obwerezabwereza m'banja la mpiru GRF. Mzere wa imvi kumbuyo ukuyimira chipika cholumikizidwa mumtundu wa mpiru, mzere wofiira umayimira kubwereza kwa magawo awiri a jini ya BjGRF;
Mawonekedwe a jini a BjGRF pansi pa zovuta za chilala pa tsamba lachinayi. Zambiri za qRT-PCR zikuwonetsedwa mu Supplementary Table S5. Kusiyana kwakukulu kwa deta kumasonyezedwa ndi zilembo zazing'ono.
Pamene nyengo yapadziko lonse ikusintha, kuphunzira momwe mbewu zimalimbana ndi chilala komanso kukonza njira zololera zakhala mutu wovuta kwambiri wofufuza18. Pambuyo pa chilala, mawonekedwe a morphological, mawonekedwe a jini ndi kagayidwe kachakudya ka zomera zidzasintha, zomwe zingayambitse kutha kwa photosynthesis ndi kusokonezeka kwa metabolic, zomwe zimakhudza zokolola ndi ubwino wa mbewu19,20,21. Zomera zikazindikira chilala, zimatulutsa amithenga achiwiri monga Ca2+ ndi phosphatidylinositol, kuonjezera intracellular calcium ion concentration ndi kuyambitsa ndondomeko yoyendetsera mapuloteni a phosphorylation pathway22,23. Mapuloteni omaliza amakhudzidwa mwachindunji ndi chitetezo cha ma cell kapena amawongolera mafotokozedwe amtundu wopsinjika wokhudzana ndi ma TF, kukulitsa kulolerana kwa mbewu kupsinjika24,25. Chifukwa chake, ma TF amagwira ntchito yofunika kwambiri pothana ndi vuto la chilala. Malinga ndi katsatidwe kake ndi DNA yomangirira katundu wa ma TF omwe amakhudzidwa ndi chilala, ma TF amatha kugawidwa m'mabanja osiyanasiyana, monga GRF, ERF, MYB, WRKY ndi mabanja ena26.
Banja la jini la GRF ndi mtundu wa TF wokhudzana ndi mbewu womwe umagwira ntchito zofunika m'mbali zosiyanasiyana monga kukula, chitukuko, kutulutsa mazizindikiro ndi mayankho oteteza mbewu27. Popeza kuti jini yoyamba ya GRF inadziwika mu O. sativa28, mitundu yambiri ya GRF yadziwika mu mitundu yambiri ya zamoyo ndikuwonetsa kuti imakhudza kukula kwa zomera, chitukuko ndi kupsinjika maganizo8, 29, 30,31,32. Ndi kusindikizidwa kwa mndandanda wa ma genome a Brassica juncea, chizindikiritso cha banja la jini la BjGRF chinatheka33. Mu kafukufukuyu, majini 34 a BjGRF adadziwika mumtundu wonse wa mpiru ndipo adatchedwa BjGRF01-BjGRF34 kutengera momwe alili. Zonsezi zili ndi madera otetezedwa a QLQ ndi WRC. Kufufuza kwa physicochemical properties kunasonyeza kuti kusiyana kwa nambala za amino acid ndi kulemera kwa maselo a mapuloteni a BjGRF (kupatula BjGRF28) sizinali zofunikira, kusonyeza kuti mamembala a BjGRF angakhale ndi ntchito zofanana. Kusanthula kapangidwe ka jini kunawonetsa kuti 64.7% ya jini ya BjGRF ili ndi ma exons 4, zomwe zikuwonetsa kuti mawonekedwe a jini a BjGRF amasungidwa bwino pachisinthiko, koma kuchuluka kwa ma exons mumitundu ya BjGRF10, BjGRF16, BjGRP28 ndi BjGRF29 ndi yayikulu. Kafukufuku wasonyeza kuti kuwonjezera kapena kuchotsedwa kwa ma exons kapena introns kungayambitse kusiyana kwa ma jini ndi ntchito, potero kumapanga majini atsopano34,35,36. Choncho, timaganiza kuti intron ya BjGRF inatayika panthawi ya chisinthiko, zomwe zingayambitse kusintha kwa majini. Mogwirizana ndi maphunziro omwe alipo, tinapezanso kuti chiwerengero cha introns chinali chogwirizana ndi jini. Pamene chiwerengero cha introns mu jini ndi chachikulu, jini akhoza mwamsanga kuyankha zinthu zosiyanasiyana zoipa.
Kubwerezabwereza kwa majini ndi chinthu chachikulu pakusintha kwa ma genomic ndi genetic37. Kafukufuku wokhudzana ndi izi awonetsa kuti kubwereza kwa majini sikumangowonjezera kuchuluka kwa majini a GRF, komanso kumagwira ntchito ngati njira yopangira majini atsopano kuti zithandizire mbewu kuti zizigwirizana ndi zovuta zosiyanasiyana zachilengedwe38. Chiwerengero cha ma gene 48 opangidwa ndi ma jini awiriawiri adapezeka mu kafukufukuyu, onsewo anali obwerezabwereza, kusonyeza kuti kubwereza kwa magawo ndi njira yaikulu yowonjezeretsa chiwerengero cha majini m'banjali. Zanenedwa m'mabuku kuti kubwereza kwa magawo kungalimbikitse kukulitsa kwa mamembala amtundu wa GRF ku Arabidopsis ndi sitiroberi, ndipo palibe kubwereza kwamtundu wamtundu uwu komwe kunapezeka mumtundu uliwonse27,39. Zotsatira za kafukufukuyu zimagwirizana ndi maphunziro omwe alipo kale a Arabidopsis thaliana ndi mabanja a sitiroberi, kutanthauza kuti banja la GRF likhoza kuonjezera chiwerengero cha majini ndikupanga majini atsopano kupyolera muzomera zosiyanasiyana.
Mu phunziro ili, chiwerengero cha 34 BjGRF majini adadziwika mu mpiru, omwe adagawidwa m'magulu ang'onoang'ono a 3. Ma jini awa adawonetsa malingaliro osungidwa ofanana ndi mapangidwe a jini. Kusanthula kwa Collinearity kunawonetsa magawo 48 a magawo obwerezabwereza mu mpiru. Chigawo cholimbikitsa cha BjGRF chili ndi zinthu za cis zomwe zimagwirizanitsidwa ndi kuyankhidwa kwa kuwala, kuyankha kwa mahomoni, kuyankha kupsinjika kwa chilengedwe, ndi kukula ndi chitukuko. Mawu a 34 BjGRF majini anapezeka pa siteji ya mpiru mbande (mizu, zimayambira, masamba), ndi chitsanzo chitsanzo cha 10 BjGRF majini pansi pa chilala. Zinapezeka kuti mafotokozedwe amtundu wa BjGRF pansi pa chilala anali ofanana ndipo angakhale ofanana. kuchita nawo chilala Kukakamiza malamulo. Majini a BjGRF03 ndi BjGRF32 atha kukhala ndi udindo wowongolera pazovuta za chilala, pomwe BjGRF06 ndi BjGRF23 zimagwira ntchito pamavuto a chilala monga miR396 chandamale. Ponseponse, kafukufuku wathu amapereka maziko achilengedwe kuti apezeke m'tsogolomu ntchito ya jini ya BjGRF muzomera za Brassicaceae.
Mbeu za mpiru zomwe zidagwiritsidwa ntchito poyesera izi zidaperekedwa ndi Guizhou Oil Seed Research Institute, Guizhou Academy of Agricultural Sciences. Sankhani mbewu zonse ndi kuzibzala m'nthaka (gawo: nthaka = 3:1), ndipo sonkhanitsani mizu, matsinde ndi masamba pambuyo pa siteji ya masamba anayi. Zomerazo zidathandizidwa ndi 20% PEG 6000 kutengera chilala, ndipo masamba adasonkhanitsidwa pambuyo pa 0, 3, 6, 12 ndi 24 hours. Zitsanzo zonse za zomera zinaumitsidwa nthawi yomweyo mu nayitrogeni wamadzimadzi ndiyeno nkusungidwa mufiriji -80°C kuti ayesenso.
Deta zonse zomwe zapezedwa kapena kufufuzidwa panthawi ya phunziroli zikuphatikizidwa m'nkhani yofalitsidwa ndi mafayilo owonjezera.
Nthawi yotumiza: Jan-22-2025