Difference between revisions of "Os10g0127900"
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To characterize the function of OsFbx352, transgenic rice plants were generated by constructing a RNAi construct with part of the coding sequence fragment to knock down OsFbx352 expression, as well as an overexpression construct with the OsFbx352 full-length open reading frame driven by a maize ubiquitin promoter. Both constructs were introduced into rice calluses via Agrobacterium-mediated transformation. Several transgenic lines were obtained and subsequently confirmed | To characterize the function of OsFbx352, transgenic rice plants were generated by constructing a RNAi construct with part of the coding sequence fragment to knock down OsFbx352 expression, as well as an overexpression construct with the OsFbx352 full-length open reading frame driven by a maize ubiquitin promoter. Both constructs were introduced into rice calluses via Agrobacterium-mediated transformation. Several transgenic lines were obtained and subsequently confirmed | ||
| − | by qRT-PCR. The expression of OsFbx352 in the overexpression seeds was at least 11-fold higher than that in WT seeds, while OsFbx352 transcripts in seeds of the knockdown lines were reduced by approximately 70% compared with WT seeds (Fig. 2). Among the transgenic lines, two overexpression lines (OE10, OE24) and two knockdown lines (Ri15, Ri18) were selected to study the physiological function of OsFbx352 during seed germination. | + | by qRT-PCR. The expression of OsFbx352 in the overexpression seeds was at least 11-fold higher than that in WT seeds, while OsFbx352 transcripts in seeds of the knockdown lines were reduced by approximately 70% compared with WT seeds (Fig. 2). Among the transgenic lines, two overexpression lines (OE10, OE24) and two knockdown lines (Ri15, Ri18) were selected to study the physiological function of OsFbx352 during seed germination.[[File:2.jpg]] |
| + | Fig. 2. Quantitative real-time PCR analysis of OsFbx352 expression in wild-type (WT), overexpression (OE10, OE11, OE13, OE14, and OE24), and RNAi (Ri6, Ri15, and Ri18) plants. Data are mean ± SE for three replicates. Values above columns are the changes in gene expression relative to the WT. | ||
===Evolution=== | ===Evolution=== | ||
Revision as of 02:47, 8 June 2014
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Contents
Annotated Information
Function
Please input function information here. This study identified an F-box gene, designed OsFbx352, from rice, and characterized its function during seed germination by generating transgenic rice plants with overexpression and knockdown of OsFbx352. More specifically, this study evaluated the role of OsFbx352 in seed germination with emphasis on the interactions among ABA, OsFbx352, and glucose. The results reveal that OsFbx352 was involved in the regulation of glucose-induced delay of seed germination by modulating ABA metabolism.
Expression
1.OsFbx352 encodes a F-box domain protein and is suppressed by glucose
A database search with the predicted polypeptide sequence showed that the F-box domain of OsFBX352 was related to those of the previously identified F-box proteins, including Arabidopsis SON (70% similarity), yeast CDC4 (65% similarity), rice OsDRF (67.5% similarity), and human SKP2 (57.5% similarity) (Fig. 1A). Phylogenetic analysis also revealed that OsFBX352 was closely related to Arabidopsis SON1 and UFO and to rice MAIF (Fig. 1B). These results suggest that OsFBX352 may be involved in the formation of E3 ubiquitin ligase complex.
Fig. 1. OsFbx352 contains a conserved F-box domain.(A) Alignment of the conserved F-box domain in OsFbx352 withother representative F-box-containing proteins: blue indicates residues identical to the OsFbx352 sequence; other colours indicate similar amino acids. The sequence at the bottom is the consensus sequence. (B)Phylogenetic tree analysis of the conserved F-box domain in OsFbx352 with other known F-box proteins from diverse organisms. Alignment and phylogenetic
analysis were performed using clustal x 2 and mega 5, respectively. Approximately 40 amino acids that constitute the
F-box motif in OsFBX352 were aligned with comparable regions from reported F-box proteins in humans (SKP2, BAB87202.1), yeast (CDC4, NP_116585.1), Arabidopsis (COI1, NP_565919; SON1, NP_179323; UFO, NP_564368.1; TIR1, NP_567135.1), and rice (OsMAIF, NP_001047698.1; OsDRF, NP_001052823.1).
2.Generation of transgenic rice plants with altered expression of OsFbx352
To characterize the function of OsFbx352, transgenic rice plants were generated by constructing a RNAi construct with part of the coding sequence fragment to knock down OsFbx352 expression, as well as an overexpression construct with the OsFbx352 full-length open reading frame driven by a maize ubiquitin promoter. Both constructs were introduced into rice calluses via Agrobacterium-mediated transformation. Several transgenic lines were obtained and subsequently confirmed
by qRT-PCR. The expression of OsFbx352 in the overexpression seeds was at least 11-fold higher than that in WT seeds, while OsFbx352 transcripts in seeds of the knockdown lines were reduced by approximately 70% compared with WT seeds (Fig. 2). Among the transgenic lines, two overexpression lines (OE10, OE24) and two knockdown lines (Ri15, Ri18) were selected to study the physiological function of OsFbx352 during seed germination.
Fig. 2. Quantitative real-time PCR analysis of OsFbx352 expression in wild-type (WT), overexpression (OE10, OE11, OE13, OE14, and OE24), and RNAi (Ri6, Ri15, and Ri18) plants. Data are mean ± SE for three replicates. Values above columns are the changes in gene expression relative to the WT.
Evolution
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Labs working on this gene
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References
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Structured Information
| Gene Name |
Os10g0127900 |
|---|---|
| Description |
Conserved hypothetical protein |
| Version |
NM_001070622.1 GI:115480987 GeneID:4348024 |
| Length |
8580 bp |
| Definition |
Oryza sativa Japonica Group Os10g0127900, complete gene. |
| Source |
Oryza sativa Japonica Group ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
| Chromosome | |
| Location |
Chromosome 10:1722588..1731167 |
| Sequence Coding Region |
1722947..1723168,1723268..1723423,1724688..1725077,1730916..1731026 |
| Expression | |
| Genome Context |
<gbrowseImage1> name=NC_008403:1722588..1731167 source=RiceChromosome10 preset=GeneLocation </gbrowseImage1> |
| Gene Structure |
<gbrowseImage2> name=NC_008403:1722588..1731167 source=RiceChromosome10 preset=GeneLocation </gbrowseImage2> |
| Coding Sequence |
<cdnaseq>ccgctggacatccagaaacgccgtcgagcaccttacgtcgccgccgtctgcccctgtcctgcaccgcgtcgagctccacttcgccgcgtttcgccatctcgccgtctcgcgggtttcttcgcacaacttgaagctggctgtccagcattggaagatctcttcttacatcaatgcggcgttcacgatgacgaaatctcctcccatacactgaaggttttgaccttcgatagtgtttttttctacatgccgatggacactgttgaattcaccttgctgaacaagacatctatttctcttccaagtgttacttctctgaccatatctactcccgagggtttcacacctatactaaaggacacggcatcagtagtgactgcatcagtatcagtatcagttacaatgtctagttttagatttcgttttgatgctaatgatctcggccagtatctccagagcctttccggtattacaaatttagagttcaactatcagggttcaaagttgacaatcgaaaaccatttgcaatggtgcccggaattcctcaatgttgtcaacctgactcttggtcaatggtgcctggattcaaacttctatgccctaattgtcttccttcagaactcacctagactacagaagctaactctgaaactcgaaaagagtaatatgagaataagtcgaagaatcattggcgagcttacggaaatatcgtttacatgcgagcaccttaatactgttgaagtgatatgctcggagaatgatccccaggtcatcactgtgcaagatttctttgttagcagtggcatgacctctgttcagtttcatatcaaacactggagcccgtacgcgaatgacctaccggcatttattcggtcgatatga</cdnaseq> |
| Protein Sequence |
<aaseq>PLDIQKRRRAPYVAAVCPCPAPRRAPLRRVSPSRRLAGFFAQLE AGCPALEDLFLHQCGVHDDEISSHTLKVLTFDSVFFYMPMDTVEFTLLNKTSISLPSV TSLTISTPEGFTPILKDTASVVTASVSVSVTMSSFRFRFDANDLGQYLQSLSGITNLE FNYQGSKLTIENHLQWCPEFLNVVNLTLGQWCLDSNFYALIVFLQNSPRLQKLTLKLE KSNMRISRRIIGELTEISFTCEHLNTVEVICSENDPQVITVQDFFVSSGMTSVQFHIK HWSPYANDLPAFIRSI</aaseq> |
| Gene Sequence |
<dnaseqindica>8000..8221#7745..7900#6091..6480#142..252#tgcatttcatcgaacaccttgcgatcaccctctcaccaccgctagagatcaccattgtgaggctccccggccggagctggaggttggacggagtggaggaagagccgtgctgccgccgtcgccatgggggaagccgggtagccgctggacatccagaaacgccgtcgagcaccttacgtcgccgccgtctgcccctgtcctgcaccgcgtcgagctccacttcgccgcgtttcgccatctcgccgtctcgcggtacaccctcgcgcctgtagcgtcctccatacatctaattgctagcgccggctccaaaatcggcgattagcgcccatggccgtgccgccccggccgccgtttcgtcgccggcgagctctccgtgggtaggctatcccgggagctcccctctccatatagttcgtgaaatcgaatccccataccccgtagatgctcccgcacccttcaattcggccggaggatgtccctagcaccgaagccccttgtagccgagctcgggcatggcgtcgcctgatggcgccgcgcgcccccgtctcctcgcgcgccggctgccattggcgggagattgccggagctcctccgcgcctcaccccggccgtcgccgagccgcccgccgccgtcccggctcgggccggccacgccgactgcgccgagccgcgcctctgcatcgctcggtaacgggcgagaggaagaagaagatgaggagagagagaaaccgtgtgggacccacatgtgattagcgcacagtaatcctatccctaacggtgattatcccactcaatggcaggtggggtccctatagttaaaatctaagaaaacacaattatatattttgttcctatgacaggcgggacccaatggtaattaaacaagcaagtttgaccctaagtcaaccatgccggtcccacatgtcaaccacactgggttttggataatgtttccgattaaaaataaatcatttatgaacagtgcatttttacaatttctcggattaactcaaatttgaatctttaaattagtataacttattcattttagctccgattcgagtgaaacttgaacctaaattcatctaaattcataagctttccaatggtatataactcattatttaataaaatatattaatatcaaatgattagattatggtcaacttaaaaatgtgctaataaataaaattgatattgtacagtagagtagatgtcaaataaatatatttaacatgttttgtcactgtagcaaccacataatctaagtataagtgatgtctcaaatgaatggatgtataggaaaatactagtgcttatttaatattcgatggccatataattaaattcatgacttaaaaattagttacataaagtttagcttcatattttgtaactaaaatattaatactcatagataaatctttaacttgtttatgagaaataataacagagctgggtgtgatgaaatcaatcgaatgatagatttaaccagtgcaatatcctatatagtagtatatttataatttggttgacacctattatttaacatattatttaacagaacaataaataactcatgttcggtaaataatccaattaacatattattggacttgcatgtacagtacatccatttaacacatacacagtaagtcaatcacctactgttgcacctattaaactatgaattgtttgataaacatcaaagttagataaaaacactaattataaatctagacctcgttacgaggccacgtagacaaataatctgtacctataacctcacagatctatatagagcatggatgattattaacccttttataatttataatgacaagtaaaatatgatcgtgattaaaataatgccccattggtaaaataataatataaatgattcactgtaattatttgattgatcctaaatctatccaacaacggaacctctagtatatgaccattttatttagatagatgcatgtgtagacataatccttccatagccattagctagactaaaccaccatgacatcagaaagactaagaaaccttagctcttagcttgattagaatccattagcaaacgcaagtagtatgagcaaccctaggttcgacctcaacaattatattttgtttgcgcacattggattgttgtttgaacattgatttttctcgcgtattatagtattggtgtatcgattagtcgtgaggcaacgtatgcagcttccaggaggtgaacgttgatcaatattatatcaagaataagaactatcctaagcaggcaagtcatcaccccattccctgagcatgttgatcccaattgtgaaattgttttgtttacaaataaaattgcatgcagtgatgaatatcctacttgttgttatgccatgcctttgattattgcttatctcattattcccctgtacccataattatgtatgagtccctagtcacttatgacaaatgcttagtaatgctaatttagattcatgcattctcatatttatcaaatgctttatgcttgggaaactaccttggggaggtagttgagatacggcatgtggagacatgagtgccacattgccatgacattgatgagatgatttgtgaaagaaaatgaaaactgaacaaatgttttcgactagggcggctggaggacttgggtggtgtccggaaaaggctagtgccgtccctggttagttaaggaccgagccatgaagctaagcatgaaacgacccccgtacaaccgtacttctcgtatgggtatagacctagcagagtaaatagccgagcggaggtagtatccctgcatagtggttttcttgatgtgtgtgaggcagggcgtacggtggggcagccattggtaggaccacgaggggcaggtgcctacagtggtgtcgccatcggtaggactgccatatgtaaacctaaacttaactataacttaatgcatgtgtgagtcttcccttcccgggagagccagaactcctctcactgctagaaaccttggacgcctagagtgcatgaggactaaagttcatggagcgggtactgccaaagtgaggttattgaaaggctttgccatgacgcgtctcatgtgttgggatgaggctcatgtgttgggcagtcacggagtatgggtaaagtgtacttccactgcagtgtgagtaaaccaatctattcgaatagccgtgctcacggttattgagcaccgggacatgtattacacttggctagactctaaattcttaaaatgatgggtgggatgttgcatgatgattttgtactggtggaacaacttctcgagaggcgagaagaggtagaccccagaaaaccatgacgactagtggcgggaagctatccttgggaacacaatggatggtggacagaaccgtcgctgtttaatgaacaatggaattaaaacttgaccagtttatgctaggtcttaggtttgtgaaatgatatttgaaaactcagctttacgcaaagtaaccataaccactccttgtattatcctgtacattactgcatttttttgtggtggcttgctgagtacttttgtactcattgttgctctatatatatcttttagcagagtgttgaagagaagcccttgttagtacgcttgcgtacctaacaagatgatcggagtgtggttccattctaggtctcgttcccagtcgactgcttgtggcatgttaaccgggcccttgtattattttgtctttcgctgttgttctctgatagttatcggcctgcctggccctaatgtaagcatataactcttttagcctgaattcattcgtgatatgttgtgattcggctgtgtgtaccaactactgatccagggattggtactgataaacacagaagatttccgatttccaaaatcggggatcgacaattactaaatctaaatctatagttatgtaatacttcactttaaattagtagtcacgtgataaatttaatattaaacatgtagttttattatatatcatcttgaataaatatatagtcttgtaatagtttgatcataggtttattattttactcaaattttcaaaatgttaaaaaataagttccaacctaattgcataacttcatatcaaatcgctaaaagcaaaaaggaacacatggtatttgagcaagtactataaaatttctaaatactagtcataagataccaatcaagcttaaatggtaatatggaggagaggagagaagaaagaaatgagagtgaaatagagaagttaattggtgacaaatgcgttgaggttagtgtaatagaggtaatatattgtataggttatttttaaatttatgagaagatgacgtggataaaacttgcttgaaccaacttggggatggtgttttcaagcaggaatcctatctcgcgcagatgcgcgagcaaactaactcggatctactaggctcaccctaccctacactgcagttgttagcattttttttctctttttcgagtgatttcctctccttccgtcgttttcccgtgcaactttttgtctctcgtgattttttttctttttctcgcgcaattttttccttttttttttgcaagattattttttcccgtgtgctatcaggctaagtttcaacttcattttattttgaaaatttttgactcaaaatttaaaaacttctaacataaaattgaaaagttctaactcaaatttagaaattttcaactgaaaatttgaaaatttcgattaagatttgaaaactttcaactaagatttgaaactttcaatttaagatttgaaaaaattcaactcaaaattgaaaacttgcaaatcaaaatttgaaaactttcaaatctaatttgaaaactttaaagttaaaaattcaaaactttcaaatcaaatttgaaaactttcaactcaaaattttaaaaaaccatatccatttttttaaactttcaagtccaaatttaaaactctcagttcatcttttaaaattttaattcaaccagtctgttcgttattttgctataaataatcacgcttatgctaatctaatcgcgtttagcgctaagcgaggggaggggcgcgtctgcgtgtggcatggcgccgtgcgcgcgagttactgtagcagcgcggatgcgcgagttacgagagaagcggaaaactgaaaacgcgcgagatctcgcgtattactccttgtcacggacagtttgagagcgggaggcctcctcccaatctccccccttccccttccatagttccatggtggcgctcttcagacgcgagcgcgaggagcaccgggtccgcttgccggtggccggcctgactcgttagccgccgtcccgtccggggcgaaaggaagcatacctcgctcactccatttggaggtaatggcttcacccaatctttcttgaatcaccattcctctctggatggatgttcttcaccaaacaatgctgtattgccgctcaatgtagcatcgtttcttgcgctaagatcttgaatcttgcttggagttgagagctggggattcatggtttattgttccgttactgtgcagcttcccccatccgcacgcgcaccagctgttcgacgaaatgccaccgcgcgctctgcccccagtggaggacgacgccggcagggactggctcggtgacctccccgaggaggtcctccaccacatcatgtccttcctagacgcgcgccaggccgtgcgaacctgtgtgctgtcgcggcggtggcgcaacctctggcgtaccgtgccttgcatcaacgctgacttcgatgagtttgatctcgtcttttaccaaggggacgacgaggactatgatgacgtgctggcgttcaaaaggtttgttaaccgtctgctggagctccgcgatcccaccgccatgatagacaccttctggcttaggtacaccaccagaccagaaggaaacacgaattccaatgaagatgcctacgggtggattagccacgctttgcagaagcaggcccgggttttagaggttgtcgtgttctgctgcttatttgagctggaccactcggtgtttacttcctgctacctgagaagaattgcgttctccggtattgttttatgcaagggtttcttcgcacaacttgaagctggctgtccagcattggaagatctcttcttacatcaatgcggcgttcacgatgacgaaatctcctcccatacactgaaggttttgaccttcgatagtgtttttttctacatgccgatggacactgttgaattcaccttgctgaacaagacatctatttctcttccaagtgttacttctctgaccatatctactcccgagggtttcacacctatactaaaggacacggcatcagtagtgactgcatcagtatcagtatcagttacaatgtctagttttagatttcgttttgatgctaatgatctcggccagtatctccagagcctttccggtattacaaatttagagttcaactatcagggttcaaaggtatgcatatttctattacatagtatgcatctttatttaggtgtcaatttgaaaaacttaaagttaagcatggtgttgtttcttcttgctacttcataattgcaattgtttaattacctgtacctgcaaaacttatgggcattgtttggataatgggatagggaaatgatttcccaactatcaaaaggtatctttaaatcctaaccatttattctccctcttccatttaatcctaacccttcattcatttcctaatcccaatcccacccttcatttcccattatccaaacacacccatgattcaagtgatcattttgtcgtgaaatcatgatgtgttgatacgatgaccactgatgttatcaaccactcaatgcaatatagttcatccactccagccattttttgtacacaaatcaatccgttgaacacttgtcctgctgtttctttgttggtagttttatctcaaaatgaaactggatcatttctcatgtgaggactagaatgttctagtctggatgcaatatcagggctgccaaccaaatgtgcatcttgtttgacatctgactgattattttagttgacatcagttcacatcttaccttactttttgaaagagggggttagcggtgagctggtgattgtaggttctagacccaaccctagtccagatttttcttggctgcagacatgtatgcattttatttctaagatatttatagaaaacaatattgcaattgcatatgcggccatttggtattgtgtaatcatgcatgaaatcgttggtttccggattaattatttattgcattttatattttagacagtatgttgaattgggatatgatgcaacgcacagacattatgcatttatgctacataaatttgaaaattaagagacattcgaaggcaatatgcaaacagaaagtgcttcaggattagttgaaaatctcacttctagtgcttcatcatgaaatatttgtactgttctttttactagtttttgttacatgacttgcatggaaagtatcctagcagtaagttaacatgcctgagactcgcattgaaaatatgaacatgtataatattgtttagtctggtcttttgttgacataatttcagtttctcctgtggtcttttaccatccgttatcctaaagttgaagctattatacatgcactccaaatagaattctttatcacatattgttaatgctctcaatttagttgccatattttctgattatattcatttgaatctttcgaagttgacaatcgaaaaccatttgcaatggtgcccggaattcctcaatgttgtcaacctgactcttggtcaatggtgcctggattcaaacttctatgccctaattgtcttccttcagaactcacctagactacagaagctaactctgaaactcgaaaaggtaaatgctttcgtccaaatgtgcactgatcctacttgattgctttttacacaagagttttcacatcgggatatagatcacctgtttttgttgctgtagagtaatatgagaataagtcgaagaatcattggcgagcttacggaaatatcgtttacatgcgagcaccttaatactgttgaagtgatatgctcggagaatgatccccaggtcatcactgtgcaagatttctttgttagcagtggcatgacctctgttcagtttcatatcaaacactggagcccgtacgcgaatgacctaccggcatttattcggtcgatatgaaatggcaaaggatcacgaattagtttgccatcccattgaagaaaatcactctctttcgcctgtcggctacatctgctggttacagtagagtgttcagtgatcttgtttggtgctttctgctatgacgtgaaggtgagtagaaacaattgcatcttgccttgcatgtaaggtcagtggtattgtacttgtttgctgcgatcacatgatgatttgtagcaataatgatgctccaagaaaaatactgtatggctatggcgtctcctgatacagcctgccactgctttctcaacttggctggtctccggtgaaacgtattgtatttgtctctctcctaatataatcccatgttatagtgttgt</dnaseqindica> 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