ʻAʻole like ka hoʻolaha ʻana o ka ua ma ke kau ma ka Panalāʻau ʻo Guizhou, me ka nui o ka ua i ka pūnāwai a me ke kauwela, akā hiki i nā hua rapeseed ke pilikia i ka maloʻo i ka hāʻule a me ka hoʻoilo, kahi e hoʻopilikia nui ai i ka hua. He mea kanu aila kūikawā ka sinapi i ulu nui ʻia ma ka Panalāʻau ʻo Guizhou. Loaʻa iā ia ka hoʻomanawanui maloʻo ikaika a hiki ke ulu ʻia ma nā wahi mauna. He waiwai nui ia o nā genes kūpaʻa maloʻo. ʻO ka ʻike ʻana i nā genes kūpaʻa maloʻo he mea nui loa ia no ka hoʻomaikaʻi ʻana i nā ʻano sinapi. a me ka hana hou i nā kumuwaiwai germplasm. He kuleana koʻikoʻi ka ʻohana GRF i ka ulu ʻana a me ka hoʻomohala ʻana o nā mea kanu a me ka pane ʻana i ke kaumaha maloʻo. I kēia manawa, ua loaʻa nā genes GRF ma Arabidopsis 2, laiki (Oryza sativa) 12, rapeseed 13, pulupulu (Gossypium hirsutum) 14, palaoa (Triticum). aestivum)15, millet pearl (Setaria italica)16 a me Brassica17, akā ʻaʻohe hōʻike o nā genes GRF i ʻike ʻia i ka sinapi. Ma kēia haʻawina, ua ʻike ʻia nā genes ʻohana GRF o ka sinapi ma ka pae genome-ākea a ua kālailai ʻia ko lākou mau ʻano kino a me ke kemika, nā pilina evolutionary, homology, nā motif i mālama ʻia, ka hoʻonohonoho ʻana o ka gene, nā duplications gene, nā cis-elements a me ke kahua seedling (kahua ʻehā lau). Ua kālailai piha ʻia nā ʻano hōʻike ma lalo o ke kaumaha maloʻo e hāʻawi i kahi kumu ʻepekema no nā haʻawina hou aʻe e pili ana i ka hana hiki o nā genes BjGRF i ka pane maloʻo a e hāʻawi i nā genes moho no ka hoʻoulu ʻana i ka sinapi hoʻomanawanui maloʻo.
Ua ʻike ʻia he kanakolukūmāhā mau genes BjGRF i loko o ka genome Brassica juncea me ka hoʻohana ʻana i ʻelua mau hulina HMMER, nona nā kikowaena QLQ a me WRC. Hōʻike ʻia nā moʻo CDS o nā genes BjGRF i ʻike ʻia ma ka Papa Hoʻohui S1. Ua kapa ʻia ʻo BjGRF01–BjGRF34 ma muli o ko lākou wahi ma ka chromosome. Hōʻike nā waiwai physicochemical o kēia ʻohana he loli nui ka lōʻihi o ka waikawa amino, mai 261 aa (BjGRF19) a i 905 aa (BjGRF28). ʻO ke kiko isoelectric o BjGRF mai 6.19 (BjGRF02) a i 9.35 (BjGRF03) me ka awelika o 8.33, a ʻo 88.24% o BjGRF he protein kumu. ʻO ka laulā kaumaha molekala i wānana ʻia o BjGRF mai 29.82 kDa (BjGRF19) a i 102.90 kDa (BjGRF28); ʻO ka helu kuhi paʻa ʻole o nā protein BjGRF mai 51.13 (BjGRF08) a i 78.24 (BjGRF19), ʻoi aku ka nui o nā mea a pau ma mua o 40, e hōʻike ana i ka helu kuhi momona mai 43.65 (BjGRF01) a i 78.78 (BjGRF22), ʻo ka awelika hydrophilicity (GRAVY) mai -1.07 (BjGRF31) a i -0.45 (BjGRF22), loaʻa i nā protein BjGRF hydrophilic nā waiwai GRAVY maikaʻi ʻole, hiki ke kumu o ka nele o ka hydrophobicity i hoʻokumu ʻia e nā koena. Ua hōʻike ka wānana o ka localization subcellular e hiki ke hoʻokomo ʻia he 31 mau protein i hoʻopili ʻia e BjGRF i loko o ka nucleus, hiki ke hoʻokomo ʻia ʻo BjGRF04 i loko o nā peroxisomes, hiki ke hoʻokomo ʻia ʻo BjGRF25 i loko o ka cytoplasm, a hiki ke hoʻokomo ʻia ʻo BjGRF28 i loko o nā chloroplasts (Papa 1), e hōʻike ana e hiki ke hoʻokomo ʻia nā BjGRF i loko o ka nucleus a hoʻokani i kahi kuleana hoʻoponopono koʻikoʻi ma ke ʻano he kumu transcription.
Hiki i ka loiloi phylogenetic o nā ʻohana GRF i nā ʻano like ʻole ke kōkua i ke aʻo ʻana i nā hana gene. No laila, ua hoʻoiho ʻia nā moʻo waikawa amino piha o 35 rapeseed, 16 turnip, 12 laiki, 10 millet a me 9 Arabidopsis GRF a ua kūkulu ʻia kahi lāʻau phylogenetic ma muli o 34 mau genes BjGRF i ʻike ʻia (Kiʻi 1). Loaʻa i nā ʻohana liʻiliʻi ʻekolu nā helu like ʻole o nā lālā; Ua māhele ʻia he 116 GRF TF i ʻekolu mau ʻohana liʻiliʻi like ʻole (nā hui A~C), nona 59 (50.86%), 34 (29.31%) a me 23 (19.83)% o nā GRF, kēlā me kēia. I waena o lākou, ua hoʻopuehu ʻia he 34 mau lālā o ka ʻohana BjGRF ma 3 mau ʻohana liʻiliʻi: 13 mau lālā ma ka hui A (38.24%), 12 mau lālā ma ka hui B (35.29%) a me 9 mau lālā ma ka hui C (26.47%). I ke kaʻina hana o ka polyploidization mustard, ʻokoʻa ka nui o nā genes BjGRFs i nā ʻohana liʻiliʻi like ʻole, a ua hiki ke hana ʻia ka hoʻonui ʻana a me ka nalowale ʻana o nā genes. He mea kūpono ke hoʻomaopopo ʻana ʻaʻohe hoʻolaha ʻana o ka laiki a me ka millet GRFs i ka hui C, ʻoiai he 2 mau GRF laiki a me 1 millet GRF i ka hui B, a ʻo ka hapa nui o nā GRF laiki a me ka millet ua hui pū ʻia i hoʻokahi lālā, e hōʻike ana he pili loa nā BjGRFs i nā dicots. I waena o lākou, ʻo nā haʻawina hohonu loa e pili ana i ka hana GRF ma Arabidopsis thaliana e hāʻawi i ke kumu no nā haʻawina hana o BjGRFs.
Lāʻau phylogenetic o ka sinapi e komo pū ana me Brassica napus, Brassica napus, laiki, millet a me nā lālā o ka ʻohana Arabidopsis thaliana GRF.
Ka nānā ʻana i nā genes hana hou ʻia i loko o ka ʻohana mustard GRF. Hōʻike ka laina hina ma ke kua i kahi poloka i hoʻonohonoho ʻia i loko o ka genome mustard, hōʻike ka laina ʻulaʻula i kahi pālua o nā hana hou i ʻāpana ʻia o ka gene BjGRF;
Ka hōʻike ʻana o ka gene BjGRF ma lalo o ke kaumaha maloʻo ma ka pae lau ʻehā. Hōʻike ʻia nā ʻikepili qRT-PCR ma ka Papa Hoʻohui S5. Hōʻike ʻia nā ʻokoʻa koʻikoʻi o ka ʻikepili e nā leka liʻiliʻi.
I ka hoʻomau ʻana o ke aniau honua i ka loli, ua lilo ke aʻo ʻana i ke ʻano o ka hoʻoponopono ʻana o nā mea kanu i ke kaumaha maloʻo a me ka hoʻomaikaʻi ʻana i kā lākou mau ʻano hoʻomanawanui i kumuhana noiʻi wela18. Ma hope o ka maloʻo, e loli ke ʻano morphological, ka hōʻike ʻana o ka gene a me nā kaʻina hana metabolic o nā mea kanu, kahi e alakaʻi ai i ka hoʻōki ʻana o ka photosynthesis a me ka hoʻopilikia metabolic, e hoʻopilikia ana i ka hua a me ka maikaʻi o nā mea kanu19,20,21. Ke ʻike nā mea kanu i nā hōʻailona maloʻo, hana lākou i nā ʻelele lua e like me Ca2+ a me phosphatidylinositol, hoʻonui i ka nui o ka ion calcium intracellular a hoʻāla i ka pūnaewele hoʻoponopono o ke ala phosphorylation protein22,23. Pili pololei ka protein target hope loa i ka pale cellular a hoʻoponopono paha i ka hōʻike ʻana o nā genes stress pili ma o TFs, e hoʻonui ana i ka hoʻomanawanui o nā mea kanu i ke kaumaha24,25. No laila, he kuleana koʻikoʻi ko TFs i ka pane ʻana i ke kaumaha maloʻo. Wahi a ke kaʻina a me nā waiwai hoʻopaʻa DNA o nā TFs pane i ke kaumaha maloʻo, hiki ke hoʻokaʻawale ʻia nā TFs i nā ʻohana like ʻole, e like me GRF, ERF, MYB, WRKY a me nā ʻohana ʻē aʻe26.
ʻO ka ʻohana gene GRF kahi ʻano TF kikoʻī o nā mea kanu e pāʻani ana i nā kuleana koʻikoʻi ma nā ʻano like ʻole e like me ka ulu ʻana, ka hoʻomohala ʻana, ka hoʻoili ʻana i nā hōʻailona a me nā pane pale o nā mea kanu27. Mai ka wā i ʻike ʻia ai ka gene GRF mua ma O. sativa28, ua ʻike ʻia nā gene GRF hou aʻe i nā ʻano he nui a ua hōʻike ʻia e hoʻopilikia i ka ulu ʻana o nā mea kanu, ka hoʻomohala ʻana a me ka pane ʻana i ke kaumaha8, 29, 30,31,32. Me ka paʻi ʻia ʻana o ka moʻo genome Brassica juncea, ua hiki ke ʻike ʻia ka ʻohana gene BjGRF33. Ma kēia haʻawina, ua ʻike ʻia he 34 mau gene BjGRF i loko o ka genome mustard holoʻokoʻa a kapa ʻia ʻo BjGRF01-BjGRF34 ma muli o ko lākou kūlana chromosomal. Loaʻa iā lākou āpau nā kikowaena QLQ a me WRC i mālama nui ʻia. Ua hōʻike ka nānā ʻana i nā waiwai physicochemical ʻaʻole koʻikoʻi nā ʻokoʻa o nā helu amino acid a me nā kaumaha molekala o nā protein BjGRF (koe wale nō ʻo BjGRF28), e hōʻike ana he like nā hana o nā lālā o ka ʻohana BjGRF. Ua hōʻike ʻia ka loiloi ʻano gene he 64.7% o nā genes BjGRF he 4 exons, e hōʻike ana ua mālama ʻia ke ʻano o ka gene BjGRF i ka evolution, akā ʻoi aku ka nui o nā exons i loko o nā genes BjGRF10, BjGRF16, BjGRP28 a me BjGRF29. Ua hōʻike ʻia nā haʻawina e hiki i ka hoʻohui ʻana a i ʻole ka holoi ʻana o nā exons a i ʻole introns ke alakaʻi i nā ʻokoʻa i ke ʻano a me ka hana o ka gene, a laila e hana ana i nā genes hou34,35,36. No laila, ke kuhi nei mākou ua nalowale ka intron o BjGRF i ka wā o ka evolution, kahi e hiki ai ke hoʻololi i ka hana o ka gene. E like me nā haʻawina e kū nei, ua ʻike pū mākou ua pili ka helu o nā introns me ka hōʻike ʻana o ka gene. Ke nui ka helu o nā introns i loko o kahi gene, hiki i ka gene ke pane koke i nā mea maikaʻi ʻole like ʻole.
ʻO ka pālua ʻana o nā genes kahi kumu nui i ka ulu ʻana o ka genomic a me ka genetic37. Ua hōʻike ʻia nā haʻawina pili ʻaʻole wale ka pālua ʻana o nā genes e hoʻonui i ka helu o nā genes GRF, akā lawelawe pū kekahi ma ke ʻano he ala e hana ai i nā genes hou e kōkua i nā mea kanu e hoʻololi i nā kūlana kaiapuni maikaʻi ʻole38. Ua loaʻa he 48 mau hui genes pālua i loko o kēia haʻawina, ʻo ia mau mea āpau he mau pālua segmental, e hōʻike ana ʻo nā pālua segmental ke ʻano nui no ka hoʻonui ʻana i ka helu o nā genes i loko o kēia ʻohana. Ua hōʻike ʻia ma ka palapala e hiki i ka pālua segmental ke hoʻolaha pono i ka hoʻonui ʻana o nā lālā o ka ʻohana gene GRF ma Arabidopsis a me ka strawberry, a ʻaʻohe pālua tandem o kēia ʻohana gene i loaʻa i kekahi o nā ʻano27,39. Kūlike nā hopena o kēia haʻawina me nā haʻawina e kū nei ma nā ʻohana Arabidopsis thaliana a me nā strawberry, e hōʻike ana e hiki i ka ʻohana GRF ke hoʻonui i ka helu o nā genes a hana i nā genes hou ma o ka pālua segmental i nā mea kanu like ʻole.
Ma kēia haʻawina, ua ʻike ʻia he 34 mau genes BjGRF i loko o ka sinapi, i māhele ʻia i 3 mau ʻohana liʻiliʻi. Ua hōʻike kēia mau genes i nā motif i mālama like ʻia a me nā ʻano genes. Ua hōʻike ka loiloi collinearity i 48 mau pālua o nā pālua ʻāpana i loko o ka sinapi. Loaʻa i ka ʻāpana hoʻolaha BjGRF nā mea cis-acting e pili ana i ka pane ʻana o ka mālamalama, ka pane hormonal, ka pane koʻikoʻi o ke kaiapuni, a me ka ulu ʻana a me ka ulu ʻana. Ua ʻike ʻia ka hōʻike ʻana o 34 mau genes BjGRF ma ke kahua kanu sinapi (aʻa, kumu, lau), a me ke ʻano hōʻike o 10 mau genes BjGRF ma lalo o nā kūlana maloʻo. Ua ʻike ʻia ua like nā ʻano hōʻike o nā genes BjGRF ma lalo o ke koʻikoʻi maloʻo a like paha. komo ʻana i ka hoʻoponopono ʻana i ka maloʻo. Hiki i nā genes BjGRF03 a me BjGRF32 ke pāʻani i nā kuleana hoʻoponopono maikaʻi i ke koʻikoʻi maloʻo, ʻoiai ʻo BjGRF06 a me BjGRF23 ke pāʻani i nā kuleana i ke koʻikoʻi maloʻo ma ke ʻano he mau genes target miR396. Ma keʻano holoʻokoʻa, hāʻawi kā mākou haʻawina i kahi kumu olaola no ka ʻike ʻana i ka hana gene BjGRF i nā mea kanu Brassicaceae i ka wā e hiki mai ana.
Ua hāʻawi ʻia nā hua sinapi i hoʻohana ʻia ma kēia hoʻokolohua e ka Guizhou Oil Seed Research Institute, Guizhou Academy of Agricultural Sciences. E koho i nā hua holoʻokoʻa a kanu iā lākou i ka lepo (substrate: lepo = 3:1), a hōʻiliʻili i nā aʻa, nā kumu a me nā lau ma hope o ke kahua lau ʻehā. Ua mālama ʻia nā mea kanu me 20% PEG 6000 e hoʻohālike i ka maloʻo, a ua hōʻiliʻili ʻia nā lau ma hope o 0, 3, 6, 12 a me 24 mau hola. Ua hoʻopaʻahau koke ʻia nā laʻana mea kanu a pau i loko o ka naikokene wai a laila mālama ʻia i loko o ka pahu hau -80°C no ka hoʻāʻo aʻe.
Hoʻokomo ʻia nā ʻikepili āpau i loaʻa a i kālailai ʻia paha i ka wā o kēia haʻawina i loko o ka ʻatikala i paʻi ʻia a me nā faila ʻike keu.
Ka manawa hoʻouna: Ian-22-2025