ʻAʻole kūlike ka hāʻawi ʻana i ka ua ma Guizhou Province, me ka nui o ka ua i ka pūnāwai a me ke kauwela, akā hiki i nā mea kanu rapeseed ke pilikia i ke koʻikoʻi maloʻo i ka hāʻule a me ka hoʻoilo, e hoʻopilikia nui i ka hua. ʻO Mustard kahi hua aila kūikawā i ulu nui ʻia ma Guizhou Province. Hiki ke ulu i nā wahi mauna. He kumu waiwai ia o nā genes kū'ē i ka maloʻo. He mea koʻikoʻi ka loaʻa ʻana o nā genes pale maloʻo no ka hoʻomaikaʻi ʻana i nā ʻano ʻano sinapi. a me ka hana hou i nā kumuwaiwai germplasm. He kuleana koʻikoʻi ka ʻohana GRF i ka ulu ʻana a me ka ulu ʻana o nā mea kanu a me ka pane ʻana i ke koʻikoʻi maloʻo. I kēia manawa, ua loaʻa nā genes GRF ma Arabidopsis 2, raiki (Oryza sativa) 12, rapeseed 13, pulupulu (Gossypium hirsutum) 14, palaoa (Triticum). aestivum)15, pearl millet (Setaria italica)16 a me Brassica17, akā ʻaʻohe hōʻike o nā genes GRF i ʻike ʻia ma ka sinapi. I loko o kēia noiʻi ʻana, ua ʻike ʻia nā genes ʻohana GRF o ka sinapi ma ka pae genome-ākea a me ko lākou mau ʻano kino a me nā ʻano kemika, nā pilina evolutionary, homology, nā kumu i mālama ʻia, ka ʻano gene, ka hoʻololi ʻana o nā gene, nā cis-elements a me ke kahua o nā mea kanu (pae ʻehā lau). ʻO nā hiʻohiʻona hōʻike ma lalo o ke koʻikoʻi maloʻo ua hoʻopaʻa ʻia e hāʻawi i kumu ʻepekema no nā noiʻi hou aku e pili ana i ka hana kūpono o nā genes BjGRF i ka pane ʻana i ka maloʻo a me ka hāʻawi ʻana i nā genes moho no ka hoʻoulu ʻana i ka sinapi hoʻomanawanui maloʻo.
Ua ʻike ʻia nā genes BjGRF he kanakolukūmāhā ma ka genome Brassica juncea me ka hoʻohana ʻana i ʻelua hulina HMMER, aia nā mea āpau i nā kāʻei QLQ a me WRC. Hōʻike ʻia nā kaʻina CDS o nā genes BjGRF i ʻike ʻia ma ka Papa Pākuʻi 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 amino acid, mai 261 aa (BjGRF19) a i 905 aa (BjGRF28). ʻO ka helu isoelectric o BjGRF mai 6.19 (BjGRF02) a 9.35 (BjGRF03) me ka awelika o 8.33, a ʻo 88.24% o BjGRF he protein kumu. ʻO ka laulā molekala kaumaha o BjGRF mai 29.82 kDa (BjGRF19) a i 102.90 kDa (BjGRF28); ʻo ka instability index o nā protein BjGRF mai 51.13 (BjGRF08) a i 78.24 (BjGRF19), ʻoi aku ka nui o nā mea āpau ma mua o 40, e hōʻike ana i ka helu ʻana o ka momona momona mai 43.65 (BjGRF01) a i 78.78 (BjGRF19) (ka awelika hydro10). (BjGRF31) i ka -0.45 (BjGRF22), loaʻa i nā protein BjGRF hydrophilic nā waiwai GRAVY maikaʻi ʻole, ʻo ia paha ma muli o ka nele o ka hydrophobicity i hana ʻia e nā koena. Hōʻike ka wānana localization subcellular e hiki ke hoʻopili ʻia nā protein i hoʻopili ʻia i ka BjGRF i loko o ka nucleus, BjGRF04 hiki ke localized i peroxisomes, BjGRF25 hiki ke localized i ka cytoplasm, a me BjGRF28 hiki ke localized i loko o nā chloroplasts (Table 1), e hōʻike ana he mea nui ka BjGRFs i loko o ka mea hoʻoponopono nucleus.
Hiki ke kōkua i ke aʻo ʻana i nā hana gene. No laila, ua hoʻoiho ʻia nā kaʻina amino acid piha piha o 35 rapeseed, 16 turnip, 12 raiki, 10 millet a me 9 Arabidopsis GRFs i hoʻoiho ʻia a ua kūkulu ʻia kahi lāʻau phylogenetic ma muli o 34 i ʻike ʻia nā genes BjGRF (Fig. 1). Aia nā ʻohana ʻekolu i nā helu like ʻole o nā lālā; Ua māhele ʻia nā 116 GRF TF i ʻekolu mau ʻohana liʻiliʻi (nā pūʻulu A~C), nona ka 59 (50.86%), 34 (29.31%) a me 23 (19.83)% o nā GRF. Ma waena o lākou, hoʻopuehu ʻia nā lālā ʻohana BjGRF ma waena o 3 mau ʻohana: 13 mau lālā ma ka hui A (38.24%), 12 mau lālā ma ka hui B (35.29%) a me 9 mau lālā i ka hui C (26.47%). Ma ke kaʻina hana o ka polyploidization sinapi, ʻokoʻa ka helu o nā genes BjGRFs i nā ʻohana subfamilis like ʻole, a ua hiki mai paha ka hoʻonui ʻana a me ka nalowale. Pono e hoʻomaopopo ʻia ʻaʻohe mahele o ka laiki a me ka millet GRF i ka hui C, ʻoiai aia he 2 GRF raiki a me 1 millet GRF ma ka hui B, a ʻo ka hapa nui o nā GRF raiki a me ka millet GRF i hui pū ʻia i kahi lālā, e hōʻike ana he pili pili nā BjGRF i nā dicots. Ma waena o lākou, nā haʻawina hohonu loa e pili ana i ka hana GRF ma Arabidopsis thaliana e hāʻawi i kumu no nā haʻawina hana o BjGRFs.
ʻO ka lāʻau Phylogenetic o ka sinapi me Brassica napus, Brassica napus, raiki, millet a me nā lālā o ka ʻohana Arabidopsis thaliana GRF.
Ka nānā ʻana o nā genes repetitive i ka ʻohana GRF sinapi. ʻO ka laina hina ma hope e hōʻike ana i kahi poloka synchronized i loko o ka genome mustard, ʻo ka laina ʻulaʻula e hōʻike ana i kahi pālua o nā ʻāpana o ka BjGRF gene;
Hōʻike gene BjGRF ma lalo o ke koʻikoʻi maloʻo ma ka pae ʻehā o ka lau. Hōʻike ʻia ka ʻikepili qRT-PCR ma ka Papa Pākuʻi S5. Hōʻike ʻia nā ʻokoʻa nui o ka ʻikepili e nā hua liʻiliʻi.
Ke hoʻomau nei ka hoʻololi ʻana o ke ea o ka honua, ʻo ke aʻo ʻana i ke ʻano o ka hoʻokō ʻana o nā mea kanu i ke koʻikoʻi o ka maloʻo a me ka hoʻomaikaʻi ʻana i kā lākou mau hana hoʻomanawanui ua lilo ia i kumuhana noiʻi wela18. Ma hope o ka maloʻo, e hoʻololi ke ʻano morphological, hōʻike gene a me nā kaʻina metabolic o nā mea kanu, hiki ke alakaʻi i ka pau ʻana o ka photosynthesis a me ka haʻalulu metabolic, e pili ana i ka hua a me ka maikaʻi o nā mea kanu19,20,21. Ke ʻike nā mea kanu i nā hōʻailona maloʻo, hoʻopuka lākou i nā ʻelele lua e like me Ca2+ a me phosphatidylinositol, e hoʻonui i ka intracellular calcium ion concentration a hoʻāla i ka pūnaewele hoʻoponopono o ke alahele phosphorylation protein22,23. Hoʻopili pololei ka protein target hope i ka pale cellular a i ʻole hoʻoponopono i ka hōʻike ʻana o nā genes koʻikoʻi pili ma o TFs, e hoʻonui ana i ka hoʻomanawanui ʻana i ka mea kanu i ke kaumaha24,25. No laila, he kuleana koʻikoʻi nā TF i ka pane ʻana i ke koʻikoʻi maloʻo. Ma muli o ke kaʻina a me nā waiwai paʻa DNA o nā TF e pane ana i ka pilikia maloʻo, hiki ke māhele ʻia nā TF i nā ʻohana like ʻole, e like me GRF, ERF, MYB, WRKY a me nā ʻohana ʻē aʻe26.
ʻO ka ʻohana gene GRF kekahi ʻano TF kikoʻī mea kanu e pāʻani ana i nā kuleana koʻikoʻi ma nā ʻano like ʻole e like me ka ulu ʻana, ka ulu ʻana, ka hoʻololi ʻana i ka hōʻailona a me nā pane pale mea kanu27. Mai ka wā i ʻike ʻia ai ka gene GRF mua ma O. sativa28, ʻoi aku ka nui o nā genes GRF i ʻike ʻia i nā ʻano he nui a hōʻike ʻia e pili ana i ka ulu ʻana o ka mea kanu, ka ulu ʻana a me ka pane pilikia8, 29, 30,31,32. Me ka paʻi ʻana o ka Brassica juncea genome sequence, hiki ke ʻike ʻia ka ʻohana gene BjGRF33. Ma kēia noiʻi ʻana, ua ʻike ʻia nā genes BjGRF 34 i loko o ka genome mustard holoʻokoʻa a kapa ʻia ʻo BjGRF01-BjGRF34 ma muli o ko lākou kūlana chromosomal. Loaʻa iā lākou a pau nā kāʻei kapu QLQ a me WRC. Ua hōʻike ʻia ka ʻike ʻana o nā waiwai physicochemical i nā ʻokoʻa o nā helu amino acid a me nā paona molekala o nā protein BjGRF (koe naʻe ʻo BjGRF28) ʻaʻole i koʻikoʻi, e hōʻike ana i nā lālā o ka ʻohana BjGRF he mau hana like. Ua hōʻike ʻia ka hōʻike ʻana o ka hoʻolālā Gene he 64.7% o nā genes BjGRF i loaʻa nā exons 4, e hōʻike ana i ka mālama ʻia ʻana o ka ʻōnaehana gene BjGRF i ka evolution, akā ʻoi aku ka nui o nā exons i nā genes BjGRF10, BjGRF16, BjGRP28 a me BjGRF29. Ua hōʻike ʻia nā haʻawina ʻo ka hoʻohui ʻana a i ʻole ka holoi ʻana i nā exons a i ʻole nā introns hiki ke alakaʻi i nā ʻokoʻa i ka hoʻolālā gene a me ka hana, a laila e hana ana i nā genes hou34,35,36. No laila, manaʻo mākou ua nalowale ka intron o BjGRF i ka wā e hoʻomohala ai, hiki ke hoʻololi i ka hana gene. E like me nā haʻawina e kū nei, ua ʻike pū mākou ua pili ka helu o nā introns me ka hōʻike gene. Ke nui ka helu o nā introns i loko o kahi gene, hiki i ka gene ke pane koke i nā ʻano mea maikaʻi ʻole.
He kumu nui ka hoʻololi ʻana i ke ʻano o ka genomic a me ka genetic evolution37. Ua hōʻike ʻia nā haʻawina e pili ana ʻaʻole e hoʻonui wale ka hoʻopili ʻana i nā genes i ka helu o nā genes GRF, akā hana pū kekahi i mea e hana ai i nā genes hou e kōkua ai i nā mea kanu e hoʻololi i nā ʻano ʻano ʻino like ʻole. Ua loaʻa he 48 mau pālua ʻano lua i loko o kēia haʻawina, ʻo ia mau mea a pau he mau ʻāpana ʻāpana, e hōʻike ana ʻo ka hoʻololi ʻana i nā ʻāpana ke kumu nui no ka hoʻonui ʻana i ka nui o nā genes i kēia ʻohana. Ua hōʻike ʻia ma ka palapala e hiki ke hoʻoikaika maikaʻi ʻia ka hoʻonui ʻana o ka ʻohana gene GRF ma Arabidopsis a me ka strawberry, a ʻaʻole i ʻike ʻia ka hoʻololi like ʻana o kēia ʻohana gene i kekahi o nā ʻano27,39. Ua kūlike nā hopena o kēia noiʻi me nā haʻawina e pili ana i nā ʻohana Arabidopsis thaliana a me nā ʻohana strawberry, e hōʻike ana e hiki i ka ʻohana GRF ke hoʻonui i ka helu o nā genes a hoʻohua i nā genes hou ma o ka hoʻopili ʻana i nā māhele i nā mea kanu like ʻole.
I loko o kēia haʻawina, ua ʻike ʻia he 34 BjGRF genes i ka sinapi, i māhele ʻia i 3 subfamilies. Hōʻike kēia mau genes i nā kumu i mālama ʻia a me nā ʻano hana gene. Ua hōʻike ʻia ka hōʻike ʻana o Collinearity he 48 mau pālua o nā ʻāpana ʻāpana i ka sinapi. Aia ka māhele hoʻolaha o BjGRF i nā mea hana cis e pili ana i ka pane māmā, ka pane hormonal, ka pane pilikia o ka nohona, a me ka ulu a me ka ulu ʻana. Ua ʻike ʻia ka hōʻike ʻana o 34 BjGRF genes i ka wā kanu hua sinapi (nā aʻa, nā ʻōpala, nā lau), a me ke ʻano hōʻike o 10 BjGRF genes ma lalo o nā kūlana maloʻo. Ua ʻike ʻia ʻo nā hiʻohiʻona hōʻike o nā genes BjGRF ma lalo o ke koʻikoʻi maloʻo ua like a like paha. komo i loko o ka maloʻo hoʻoponopono hoʻoponopono. Hiki i nā genes BjGRF03 a me BjGRF32 ke pāʻani i nā kuleana hoʻoponopono maikaʻi i ke koʻikoʻi o ka maloʻo, ʻoiai ʻo BjGRF06 a me BjGRF23 e pāʻani i ke koʻikoʻi o ka maloʻo e like me nā genes target miR396. ʻO ka holoʻokoʻa, hāʻawi kā mākou noiʻi i kumu olaola no ka ʻike ʻana o ka hana gene BjGRF i nā mea kanu Brassicaceae.
Hāʻawi ʻia nā hua sinapi i hoʻohana ʻia i kēia hoʻokolohua e Guizhou Oil Seed Research Institute, Guizhou Academy of Agricultural Sciences. E koho i nā ʻanoʻano holoʻokoʻa a kanu i ka lepo (substrate: soil = 3:1), a e ʻohi i nā aʻa, nā ʻōpala a me nā lau ma hope o ka pae ʻehā-lau. Ua mālama ʻia nā mea kanu me 20% PEG 6000 e hoʻohālikelike i ka maloʻo, a ʻohi ʻia nā lau ma hope o 0, 3, 6, 12 a me 24 mau hola. Ua maloo koke nā laʻana mea kanu i loko o ka hau hau a mālama ʻia i loko o ka pahu hau -80°C no ka hoʻāʻo aʻe.
Hoʻokomo ʻia nā ʻikepili āpau i loaʻa a i ʻike ʻia i ka wā o kēia noiʻi ʻana i ka ʻatikala i paʻi ʻia a me nā faila ʻike hou.
Ka manawa hoʻouna: Jan-22-2025