Os09g0422500

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Annotated Information

Function

1;This study characterizes a brittle culm (bc88) mutant of rice (Oryza sativa L.) obtained by ethylene methylsulfonate (EMS)-induced mutagenesis of Wuyunjing 7. The bc88 mutant exhibits a diversity of pleiotropic phenotypes, including brittle culm at the whole-plant growth stages, withered leaf tips at the seedling stage, and 18-d delay in heading date at the mature stage. Genetic analysis indicates that the bc88 mutant is controlled by a single recessive nuclear gene. The mutated bc88 gene isolated by map-based cloning contains only one point mutation in the 5th exon relative to its wild-type BC88 (LOC_Os09g25490 and Os09g0422500), leading to an amino acid change from P to L in bc88 plants. Alignment of the putative protein sequence with its homologs indicates that the mutation is located in the conserved region of the sequence. Detection of the transcription level of BC88 in rice plants shows that the expression level of BC88 is higher in spikes and culms than in leaves, roots, and leaf sheaths. These contribute to understanding of the molecular mechanism of cellulose synthesis. The target gene BC88 can be a useful tool in molecular marker-assisted selection for rice culm trait breeding.

2;Rice is a model organism in poaceae plants to study cell wall biosynthesis. In this study, a mutant S1-60 isolated from an EMS mutagenized japonica cultivar Nipponbare, is characterized by brittle culms that can be easily broken by bending. The reduction in mechanical strength was due to defect in thickening of the sclerenchyma cell wall. The amount of cellulose in S1-60 culms was reduced to 44.7% of that of wild-type plants. Besides, the mutant also exhibited pleiotropic phenotypes, such as dwarfism and partial sterility. Genetic analysis and map-based cloning showed that all the phenotype of S1-60 mutant was caused by a recessive point mutation in the OsCESA9 gene, which encodes the cellulose synthase A subunit 9. This yet uncharacterized missense mutation changed the highly conserved G905 to D at the beginning of the fifth transmembrane domain. The OsCESA9 gene is predominantly expressed in the culms of mature stage plants, consistent with the brittle phenotype in the culm. These results indicate that OsCESA9 plays an important role in cell wall biosynthesis and plant growth.

3;According to the MSU Rice Genome Annotation Release 7 (http://rice.plantbiology.msu.edu), there are 13 predicted open reading frames (ORFs) within the 118 kb fine mapping interval , including 7 ORFs with known biochemical functions, 5 ORFs encoding expressed hypothetical protein and 1 transposon . Among them, ORF3 (TIGR ID: LOC_Os09g25490) encoding cellulose synthase A catalytic subunit 9 (OsCESA9) was considered as the priority candidate gene, given that the amount of cellulose was reduced dramatically in S1-60 culm. Therefore, we sequenced and compared the mutant and wild type alleles of the OsCESA9 gene, which has 4643 bp in length and 11 exons and 10 introns. One base pair substitution was found in the last exon, changing GGC to GAC and the encoded amino acid from glycine to aspartic acid at the 905th position .The CESA9 protein has 1056 amino acids in length and eight putative transmembrane domains (TMDs), with two near the amino terminus and six clustered near the carboxyl terminus . There is a RING-type zinc finger in the N-terminal region, which might mediate the interaction between CESA9 and other CESA subunits. A large central domain containing two highly conserved motifs DXD and Q/RXXRW is required for the catalytic activity of the enzyme. The missense mutation in S1-60 occurs at the beginning of the fifth TMD, which might affect the plasma membrane localization of CESA9.

Expression

We examined the expression level of OsCESA9 in various rice organs by both semi-quantitative ( Fig. 7A) and quantitative RT-PCR ( Fig. 7B). The OsCESA9 gene is predominantly expressed in the culms of mature stage plants, consistent with the brittle culm phenotype in the S1-60 mutant. The OsCESA9 gene is also expressed in panicle at mature stage. However, the expression level of OsCESA9 was relatively low at seedling stage, no matter in root, leaf blade or leaf sheath. This result showed the OsCESA9 gene mainly participates in the synthesis of secondary cell wall in mechanical tissue at late development stage.

Evolution

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Labs working on this gene

1. College of Agronomy and Plant Protection, Qingdao Agricultural University, Qingdao 266109, China

2. National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, China

3. Shandong Rice Research Institute, Jinan 250100, China

4. College of Life Sciences, Qingdao Agricultural University, Qingdao 266109, China

5. Division of Life Science, Graduate School of Science and Engineering, Saitama University, 255 Shimo-okubo, Sakura-ku, Saitama 338-8570, Japan

References

1.A missense mutation in the transmembrane domain of CESA9 affects cell wall biosynthesis and plant growth in rice. Wang D, et al. Plant Sci, 2012 Nov. PMID 23017906

2.Rice Brittle culm 6 encodes a dominant-negative form of CesA protein that perturbs cellulose synthesis in secondary cell walls. Kotake T, et al. J Exp Bot, 2011 Mar. PMID 21209026, Free PMC Article

3.The Rice Annotation Project Database (RAP-DB): 2008 update. Rice Annotation Project, et al. Nucleic Acids Res, 2008 Jan. PMID 18089549, Free PMC Article

4.Curated genome annotation of Oryza sativa ssp. japonica and comparative genome analysis with Arabidopsis thaliana. Rice Annotation Project, et al. Genome Res, 2007 Feb. PMID 17210932, Free PMC Article

5.The Rice Annotation Project Database (RAP-DB): hub for Oryza sativa ssp. japonica genome information. Ohyanagi H, et al. Nucleic Acids Res, 2006 Jan 1. PMID 16381971, Free PMC Article

Structured Information

Gene Name	Os09g0422500
Description	Similar to Cellulose synthase (Fragment)
Version	NM_001069742.1 GI:115479226 GeneID:4347093
Length	4574 bp
Definition	Oryza sativa Japonica Group Os09g0422500, 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	Chromosome 9
Location	Chromosome 9:15935031..15939604
Sequence Coding Region	15935124..15935394,15935500..15935735,15935831..15935927,15936019..15936145,15936232..15936844 ,15936934..15937197,15937280..15937492,15937615..15937822,15937914..15938110 ,15938192..15938545,15938649..15939236
Expression	GEO Profiles:Os09g0422500
Genome Context	<gbrowseImage1> name=NC_008402:15935031..15939604 source=RiceChromosome09 preset=GeneLocation </gbrowseImage1>
Gene Structure	<gbrowseImage2> name=NC_008402:15935031..15939604 source=RiceChromosome09 preset=GeneLocation </gbrowseImage2>
Coding Sequence	<cdnaseq>atggaggcgagcgccgggctggtggccgggtcgcacaaccggaacgagctggtgctgatccgggggcacgaggagcccaagccgctgcgggcgctgagcgggcaggtgtgcgagatatgcggcgacgaggtcggccgcaccgtcgacggcgacctcttcgtcgcctgcaacgagtgcggcttcccggtgtgccgcccctgctacgagtacgagcgccgcgagggcacccagaactgcccccagtgcaagacccgctacaagcgcctcaaggggagcccgagggtgcccggggacgaggacgaggaggacattgacgacctggagcacgagttcaacatcgacgacgagaagcagaagcagctgcagcaggatcaggatggcatgcagaacagccacatcaccgaggcgatgctgcacggcaagatgagctacgggaggggccccgacgacggcgacggcaacagcaccccgctcccgccgatcatcaccggcgctcgctccgtcccggtgagcggggagttcccgatatcgaacagccatggccatggcgagttctcctcttccctgcacaagcgcatccacccctacccggtgtctgagccagggagtgcaaagtgggacgagaagaaagaggtgagctggaaggagaggatggacgactggaaatccaagcagggcatcgtcgccggcggcgcccccgatcccgacgactacgacgccgacgtcccactgaacgacgaggcgaggcagccgctgtcgaggaaggtgtcgatcgcgtcgagcaaggtgaacccgtaccggatggtgatcatcctccgtctcgtggtgctcggcttcttcctccggtaccgcatcctccacccggtgcccgacgccatcccgctgtggctcacctccatcatctgcgagatctggttcgccgtgtcgtggatcctcgaccagttccccaagtggtacccgatcgacagggagacctacctcgaccgcctctccctccgctacgagcgcgagggggagccgtcgctgctgtcggcggtggacctgttcgtcagcacggtggatccgctcaaggagccgccgctggtcacggccaacaccgtgctgtccatcctcgccgtcgactaccccgtcgacaaggtgtcctgctacgtctccgacgacggcgcgtccatgctcacgttcgagtcgctgtcggagacggcggagttcgcccgcaagtgggtgccattctgcaagaagttcagcatcgagccccgcgccccggagttctacttctcccagaaggtcgactacctcaaggacaaggtccatcccaacttcgtccaggagcgccgcgccatgaagagagagtacgaggagttcaaggtgaggatcaacgcgctggtggcgaaggcgcagaaggtgccggcggaagggtggatcatgaaggacgggacgccatggccggggaacaacacccgcgaccacccgggcatgatccaggtgttcctcggccacagcggcggccacgacaccgagggcaacgagctcccccgcctcgtctacgtctcccgtgagaagcgccccggcttccagcaccacaagaaggccggcgccatgaacgccctcattcgtgtgtcggccgtgctgacgaacgcgccgttcatgctcaacttggattgcgatcactacatcaacaacagcaaggccatcagggaggcgatgtgcttcctcatggatccgcaggtcggacggaaggtttgctacgtgcagttcccgcagaggttcgacggcatcgacgtccacgaccgatacgccaaccgcaacaccgtcttcttcgacatcaacatgaaggggcttgatgggatccagggcccggtgtacgtcgggacagggtgcgtgttcaggcggcaggcgctgtacggatacaacccacccaagggacccaagaggcccaagatggtgacctgcgactgctgcccttgcttcgggaggaagaagcggaagcacggcaaggacggcctcccggaggccgtcgccgccgacggcgggatggacagcgacaaggagatgctcatgtcgcagatgaacttcgagaagcggttcgggcagtcggcggcgttcgtgacgtcgacgctgatggaggaaggcggcgtcccgccgtcgtccagccccgccgcgctcctcaaggaggccatccatgtcatcagctgcggctacgaggacaagaccgactggggtctcgagctggggtggatctacgggtcgatcacggaggacatcctaacggggttcaagatgcactgccgcgggtggaggtcggtgtactgcatgccgaagagggcggcgttcaaggggtcagcgccgatcaacctatctgaccgtctcaaccaggtgctccggtgggcgctcggctccgtcgagatcttcttcagccggcacagcccgctcctctacggctacaagaacggcaacctcaagtggctcgagcgcttctcctacatcaacaccaccatctaccccttcacttctctccccctcctcgcctactgcaccctacccgccgtctgcctcctcaccggcaagttcatcatgcctccgattagcacgtttgcgagtttgttcttcatcgcgctcttcatctccatcttcgcgacgggcatcctggagatgaggtggagcggggtgagcatcgaggagtggtggaggaacgagcagttctgggtcatcggcggcgtgtcggcgcacctgttcgcggtggtgcagggcctgctcaaggtgctggccgggatcgacaccaacttcaccgtcacgtccaaggccaccggagacgaggacgacgagttcgcggagctctacgccttcaagtggaccaccctcctcatcccgcccaccacgctgctcatcctcaacatcatcggcgtcgtcgccggcgtctccgacgccatcaacaacggctccgaggcgtggggcccgctcttcgggaagctcttcttcgccttctgggtcatcgtccacctctaccccttcctcaaggggctcatggggaggcagaaccggacgcccaccattgttgtcatctggtccgtgctgctcgcctccatcttttccttgctctgggtcaggattgatcccttcaccatcaaggccaggggccctgacgtcaggcagtgcggcatcaactgctga</cdnaseq>
Protein Sequence	<aaseq>MEASAGLVAGSHNRNELVLIRGHEEPKPLRALSGQVCEICGDEV GRTVDGDLFVACNECGFPVCRPCYEYERREGTQNCPQCKTRYKRLKGSPRVPGDEDEE DIDDLEHEFNIDDEKQKQLQQDQDGMQNSHITEAMLHGKMSYGRGPDDGDGNSTPLPP IITGARSVPVSGEFPISNSHGHGEFSSSLHKRIHPYPVSEPGSAKWDEKKEVSWKERM DDWKSKQGIVAGGAPDPDDYDADVPLNDEARQPLSRKVSIASSKVNPYRMVIILRLVV LGFFLRYRILHPVPDAIPLWLTSIICEIWFAVSWILDQFPKWYPIDRETYLDRLSLRY EREGEPSLLSAVDLFVSTVDPLKEPPLVTANTVLSILAVDYPVDKVSCYVSDDGASML TFESLSETAEFARKWVPFCKKFSIEPRAPEFYFSQKVDYLKDKVHPNFVQERRAMKRE YEEFKVRINALVAKAQKVPAEGWIMKDGTPWPGNNTRDHPGMIQVFLGHSGGHDTEGN ELPRLVYVSREKRPGFQHHKKAGAMNALIRVSAVLTNAPFMLNLDCDHYINNSKAIRE AMCFLMDPQVGRKVCYVQFPQRFDGIDVHDRYANRNTVFFDINMKGLDGIQGPVYVGT GCVFRRQALYGYNPPKGPKRPKMVTCDCCPCFGRKKRKHGKDGLPEAVAADGGMDSDK EMLMSQMNFEKRFGQSAAFVTSTLMEEGGVPPSSSPAALLKEAIHVISCGYEDKTDWG LELGWIYGSITEDILTGFKMHCRGWRSVYCMPKRAAFKGSAPINLSDRLNQVLRWALG SVEIFFSRHSPLLYGYKNGNLKWLERFSYINTTIYPFTSLPLLAYCTLPAVCLLTGKF IMPPISTFASLFFIALFISIFATGILEMRWSGVSIEEWWRNEQFWVIGGVSAHLFAVV QGLLKVLAGIDTNFTVTSKATGDEDDEFAELYAFKWTTLLIPPTTLLILNIIGVVAGV SDAINNGSEAWGPLFGKLFFAFWVIVHLYPFLKGLMGRQNRTPTIVVIWSVLLASIFS LLWVRIDPFTIKARGPDVRQCGINC</aaseq>
Gene Sequence	<dnaseqindica>94..364#470..705#801..897#989..1115#1202..1814#1904..2167#2250..2462#2585..2792#2884..3080#3162..3515#3619..4206#agcgatcgatcgcccttcctcctcctcctctcctccttcctgcgtcgcctccctgatcagccgcagctcgttgccggccgttgttgcgcggccatggaggcgagcgccgggctggtggccgggtcgcacaaccggaacgagctggtgctgatccgggggcacgaggagcccaagccgctgcgggcgctgagcgggcaggtgtgcgagatatgcggcgacgaggtcggccgcaccgtcgacggcgacctcttcgtcgcctgcaacgagtgcggcttcccggtgtgccgcccctgctacgagtacgagcgccgcgagggcacccagaactgcccccagtgcaagacccgctacaagcgcctcaagggtaaaaaaacacactcacatcacgctacgcccttgataccgcgatcgcgaggtttccgttgattgatctctaatggcgatggtggtggtggtggttttgttacagggagcccgagggtgcccggggacgaggacgaggaggacattgacgacctggagcacgagttcaacatcgacgacgagaagcagaagcagctgcagcaggatcaggatggcatgcagaacagccacatcaccgaggcgatgctgcacggcaagatgagctacgggaggggccccgacgacggcgacggcaacagcaccccgctcccgccgatcatcaccggcgctcgctccgtcccggtacacacaacacacctcaccccactcacaccaaattctctccctcttctcacctcaacatgttcacgaaatgttctttgtaaaaaaaaattcaggtgagcggggagttcccgatatcgaacagccatggccatggcgagttctcctcttccctgcacaagcgcatccacccctacccggtgtctgagccaggtagtataacgaattcactacactaattaatcaatgttcttgatgaacacacattgatcatctcaatcatctaccgatgaattctgaccagggagtgcaaagtgggacgagaagaaagaggtgagctggaaggagaggatggacgactggaaatccaagcagggcatcgtcgccggcggcgcccccgatcccgacgactacgacgccgacgtcccactgtacgcaatcctcagctgagcagcttacactgatcatgcatgacaactagtatattgatcatgcctgaactctctgtggcttgcaggaacgacgaggcgaggcagccgctgtcgaggaaggtgtcgatcgcgtcgagcaaggtgaacccgtaccggatggtgatcatcctccgtctcgtggtgctcggcttcttcctccggtaccgcatcctccacccggtgcccgacgccatcccgctgtggctcacctccatcatctgcgagatctggttcgccgtgtcgtggatcctcgaccagttccccaagtggtacccgatcgacagggagacctacctcgaccgcctctccctccgctacgagcgcgagggggagccgtcgctgctgtcggcggtggacctgttcgtcagcacggtggatccgctcaaggagccgccgctggtcacggccaacaccgtgctgtccatcctcgccgtcgactaccccgtcgacaaggtgtcctgctacgtctccgacgacggcgcgtccatgctcacgttcgagtcgctgtcggagacggcggagttcgcccgcaagtgggtgccattctgcaagaagttcagcatcgagccccgcgccccggagttctacttctcccagaaggtcgactacctcaaggacaaggtccatcccaacttcgtccaggagcgccgcgccatgaaggtaatatcgatcgccatcgtcattgctgattctgtaggagcttgacgaagagctaactgttgtgggggcattggcatttgatcgagcagagagagtacgaggagttcaaggtgaggatcaacgcgctggtggcgaaggcgcagaaggtgccggcggaagggtggatcatgaaggacgggacgccatggccggggaacaacacccgcgaccacccgggcatgatccaggtgttcctcggccacagcggcggccacgacaccgagggcaacgagctcccccgcctcgtctacgtctcccgtgagaagcgccccggcttccagcaccacaagaaggccggcgccatgaacgccctcgtacgccacgccaccacatccttcatcttcaaaacaacacaactctgctcgatttgatcgaaatttctggtttgcttggcagattcgtgtgtcggccgtgctgacgaacgcgccgttcatgctcaacttggattgcgatcactacatcaacaacagcaaggccatcagggaggcgatgtgcttcctcatggatccgcaggtcggacggaaggtttgctacgtgcagttcccgcagaggttcgacggcatcgacgtccacgaccgatacgccaaccgcaacaccgtcttcttcgacgtgagctctctcccacacaaactagatgaatatatacaatcttgaaaaatccagagcaaatcacgatcgatcgagcaatgtgactgaaattttgtgtggtgcgttcttggtgagatcatcagatcaacatgaaggggcttgatgggatccagggcccggtgtacgtcgggacagggtgcgtgttcaggcggcaggcgctgtacggatacaacccacccaagggacccaagaggcccaagatggtgacctgcgactgctgcccttgcttcgggaggaagaagcggaagcacggcaaggacggcctcccggaggccgtcgccgccgacggcggtgagctcccaaattcagagctaagaagaaattatggtgtgagaagattttcagctgatggaataatgtaagtttgtgtgtggtggatcagggatggacagcgacaaggagatgctcatgtcgcagatgaacttcgagaagcggttcgggcagtcggcggcgttcgtgacgtcgacgctgatggaggaaggcggcgtcccgccgtcgtccagccccgccgcgctcctcaaggaggccatccatgtcatcagctgcggctacgaggacaagaccgactggggtctcgaggtaaccaccgttatcagacactaagccatattaccattgagtgagtttgtggtgtgaacgccatggatgtgtgtgatgcagctggggtggatctacgggtcgatcacggaggacatcctaacggggttcaagatgcactgccgcgggtggaggtcggtgtactgcatgccgaagagggcggcgttcaaggggtcagcgccgatcaacctatctgaccgtctcaaccaggtgctccggtgggcgctcggctccgtcgagatcttcttcagccggcacagcccgctcctctacggctacaagaacggcaacctcaagtggctcgagcgcttctcctacatcaacaccaccatctaccccttcacttctctccccctcctcgcctactgcaccctacccgccgtctgcctcctcaccggcaagttcatcatgcctccggtcagtcccatcccatcctgccatcaattgctcctcttcttccatggattttcccgccaaaactgaagtttcaaatgttctgaaccttttcttggtgatgcagattagcacgtttgcgagtttgttcttcatcgcgctcttcatctccatcttcgcgacgggcatcctggagatgaggtggagcggggtgagcatcgaggagtggtggaggaacgagcagttctgggtcatcggcggcgtgtcggcgcacctgttcgcggtggtgcagggcctgctcaaggtgctggccgggatcgacaccaacttcaccgtcacgtccaaggccaccggagacgaggacgacgagttcgcggagctctacgccttcaagtggaccaccctcctcatcccgcccaccacgctgctcatcctcaacatcatcggcgtcgtcgccggcgtctccgacgccatcaacaacggctccgaggcgtggggcccgctcttcgggaagctcttcttcgccttctgggtcatcgtccacctctaccccttcctcaaggggctcatggggaggcagaaccggacgcccaccattgttgtcatctggtccgtgctgctcgcctccatcttttccttgctctgggtcaggattgatcccttcaccatcaaggccaggggccctgacgtcaggcagtgcggcatcaactgctgagagagggcgtcagcgatttctgaagatttgtgcataggggggcagcaagaagatcgatttgtaaaagttttgtattgctcgtgttgagttcgtgctatgttcttctgtaatattttgggacccagaaatggtttaaacttttgaagagggaatggacagatggccatatttgaatgttaagcaacaagggctggtattctcactccatgtttgttagattttttgcagctgtgttctcattgctcctactagggatatatgggtaatttggaccaaaccatgggcattaatgtagcttaaaagtatatgaatcgatttggtcaagggctgaagtaatattcctacaaggaatatattcagattgcagc</dnaseqindica>
External Link(s)	NCBI Gene:Os09g0422500, RefSeq:Os09g0422500