Os01g0884300

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Annotated Information

Function

 The OsNAC6 gene is a member of the NAC transcription factor gene family in rice. Expression of OsNAC6 is induced by abiotic stresses, including cold, drought and high salinity. OsNAC6 gene expression is also induced by wounding and blast disease. A transactivation assay using a yeast system demonstrated that OsNAC6 functions as a transcriptional activator, and transient localization studies with OsNAC6–sGFP fusion protein revealed its nuclear localization. Transgenic rice plants over-expressing OsNAC6 constitutively exhibited growth retardation and low reproductive yields. These transgenic rice plants showed an improved tolerance to dehydration and high-salt stresses, and also exhibited increased tolerance to blast disease. By utilizing stressinducible promoters, such as the OsNAC6 promoter, it is hoped that stress-inducible over-expression of OsNAC6 in rice can improve stress tolerance by suppressing the negative effects of OsNAC6 on growth under normal growth conditions. The results of microarray analysis revealed that many genes that are inducible by abiotic and biotic stresses were upregulated in rice plants over-expressing OsNAC6. A transient transactivation assay showed that OsNAC6 activates the expression of at least two genes, including a gene encoding peroxidase. Collectively, these results indicate that OsNAC6 functions as a transcriptional activator in response to abiotic and biotic stresses in plants. We conclude that OsNAC6 may serve as a useful biotechnological tool for the improvement of stress tolerance in various kinds of plants.
 OsNAC6 functions as a transcriptional activator and is localized in the nucleus An OsNAC6–sGFP fusion protein driven by the CaMV 35S promoter was transiently expressed in onion epidermal cells and analyzed by fluorescent microscopy. A SV40 NLS–sGFP fusion protein (SV40NLS–sGFP) and sGFP alone (35S–sGFP), driven by the 35S promoter, were used as a positive control (nuclear localization) and negative control, respectively. Nuclear localization was confirmed for OsNAC6 as both OsNAC6–sGFP and the positive control (SV40NLS–sGFP) were localized in the nucleus, whereas 35S–sGFP was localized in both cytoplasm and nucleus (Figure 2a). We also examined the transcriptional activity of OsNAC6 using a yeast system. A GAL4 DNA binding domain–OsNAC6 fusion protein was expressed in yeast cells, which were assayed for their ability to activate transcription from the GAL4 binding sequence. OsNAC6 promoted yeast growth in the absence of histidine and showed b-galactosidase activity, while the vector control pGBKT7 did not (Figure 2b). These data confirm that OsNAC6 functions as a transcriptional activator.

Expression

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 Expression of OsNAC6 is induced by both abiotic and biotic stresses
 The OsNAC6 gene (AB028185; AK068392; Os01 g0884300; ONAC048) encodes a protein of 303 amino acids containing the NAC domain in its N-terminal region (Figure 1a). The NAC domain contains predicted nuclear localization signals (NLS) at amino acids 71–83 and 107–123. RNA gel-blot and quantitative polymerase chain reaction (PCR) analyses showed that OsNAC6 was induced by dehydration, high salt (250 mM NaCl), cold (4�C), 100 lM ABA, 100 lM methyl

jasmonate (MeJA) (Figure 1b and Supplementary Figure S1) and wounding (Figure 1c). Induction of OsNAC6 was observed in leaves infected with the blast fungus Magnaporthe grisea Kyu89-246 (Figure 1d). We also examined the effects of stress-related chemicals on the expression of OsNAC6 in rice culture cells. Quantitative PCR analysis showed that OsNAC6 was moderately induced by hydrogen peroxide (H2O2) and weakly by the elictor N-acetylchitooligosaccharide (Figure 1e).

 In order to assess the effect of the promoter region on the expression of OsNAC6 under abiotic and biotic stresses in leaves and roots, we generated the transgenic rice plants containing 1.5 kb OsNAC6 promoter–GUS chimeric genes. Quantitative analysis of the OsNAC6 promoter–GUS transgenic rice plants showed that OsNAC6 was induced by dehydration, high salinity, cold, ABA, MeJA, hydrogen peroxide, wounding and blast disease (Figure 1f,g).

Sequences of various cis-acting elements involved in the response to abiotic stresses were identified in the 1.5 kb promoter region of OsNAC6 (Figure 1h). We found three ABA-responsive elements (ABREs; ACGTGG/TC) (Hattori et al., 2002), three recognition sites for MYB (MYBRSs; C/TAACNA/G) (Abe et al., 2003) and six recognition sites for MYC (MYCRSs; CANNTG) (Abe et al., 2003). The OsNAC6 promoter also includes some cis-acting elements involved in the reponse to biotic stresses, such as four W-boxes (TTGAC) (Eulgem et al., 2000) and four GCC boxes (GCCGCC) (Brown et al., 2003), which are known as recognition sites for WRKY and ERF transcription factors, respectively. Additionally, the OsNAC6 promoter has three

Evolution

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Labs working on this gene

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References

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Structured Information

Gene Name	Os01g0884300
Description	No apical meristem (NAM) protein domain containing protein
Version	NM_001051551.1 GI:115441472 GeneID:4325006
Length	2486 bp
Definition	Oryza sativa Japonica Group Os01g0884300, 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 1
Location	Chromosome 1:40154843..40157328
Sequence Coding Region	40155348..40155818,40156680..40156954,40157054..40157219
Expression	GEO Profiles:Os01g0884300
Genome Context	<gbrowseImage1> name=NC_008394:40154843..40157328 source=RiceChromosome01 preset=GeneLocation </gbrowseImage1>
Gene Structure	<gbrowseImage2> name=NC_008394:40154843..40157328 source=RiceChromosome01 preset=GeneLocation </gbrowseImage2>
Coding Sequence	<cdnaseq>atgagcggcggtcaggacctgcagctgccgccggggttccggttccacccgacggacgaggagctggtgatgcactacctctgccgccgctgcgccggcctccccatcgccgtccccatcatcgccgagatcgacctctacaagttcgatccatggcagcttccccggatggcgctgtacggagagaaggagtggtacttcttctccccgcgagaccgcaagtacccgaacgggtcgcggccgaaccgcgccgccgggtcggggtactggaaggcgaccggcgccgacaagccggtgggctcgccgaagccggtggcgatcaagaaggccctcgtcttctacgccggcaaggcgcccaagggcgagaagaccaactggatcatgcacgagtaccgcctcgccgacgtcgaccgctccgcccgcaagaagaacagcctcaggttggatgattgggtgctgtgccggatttacaacaagaagggcgggctggagaagccgccggccgcggcggtggcggcggcggggatggtgagcagcggcggcggcgtccagaggaagccgatggtgggggtgaacgcggcggtgagctccccgccggagcagaagccggtggtggcggggccggcgttcccggacctggcggcgtactacgaccggccgtcggactcgatgccgcggctgcacgccgactcgagctgctcggagcaggtgctgtcgccggagttcgcgtgcgaggtgcagagccagcccaagatcagcgagtgggagcgcaccttcgccaccgtcgggcccatcaaccccgccgcctccatcctcgaccccgccggctccggcggcctcggcggcctcggcggcggcggcagcgaccccctcctccaggacatcctcatgtactggggcaagccattctag</cdnaseq>
Protein Sequence	<aaseq>MSGGQDLQLPPGFRFHPTDEELVMHYLCRRCAGLPIAVPIIAEI DLYKFDPWQLPRMALYGEKEWYFFSPRDRKYPNGSRPNRAAGSGYWKATGADKPVGSP KPVAIKKALVFYAGKAPKGEKTNWIMHEYRLADVDRSARKKNSLRLDDWVLCRIYNKK GGLEKPPAAAVAAAGMVSSGGGVQRKPMVGVNAAVSSPPEQKPVVAGPAFPDLAAYYD RPSDSMPRLHADSSCSEQVLSPEFACEVQSQPKISEWERTFATVGPINPAASILDPAG SGGLGGLGGGGSDPLLQDILMYWGKPF</aaseq>
Gene Sequence	<dnaseqindica>1511..1981#375..649#110..275#caagccctcctctcctcttcccaacactagtaggataaagccacagagagagcagtagtagtagcgagctcgccggagaacggacgatcaccggagaagggggagagagatgagcggcggtcaggacctgcagctgccgccggggttccggttccacccgacggacgaggagctggtgatgcactacctctgccgccgctgcgccggcctccccatcgccgtccccatcatcgccgagatcgacctctacaagttcgatccatggcagcttccccgtacgataatcctcctcctccatcctcccaatcatcaccaccatcaacgccgtcgtgaattgattgattgatttggtttgatttgttggtgttgtgtagggatggcgctgtacggagagaaggagtggtacttcttctccccgcgagaccgcaagtacccgaacgggtcgcggccgaaccgcgccgccgggtcggggtactggaaggcgaccggcgccgacaagccggtgggctcgccgaagccggtggcgatcaagaaggccctcgtcttctacgccggcaaggcgcccaagggcgagaagaccaactggatcatgcacgagtaccgcctcgccgacgtcgaccgctccgcccgcaagaagaacagcctcagggtaagcaaaaaccacacccaagattccatcactaaattcattactaaatctgtgttcatcgtgattattgattaatttagtcacctaattattcgcccaaaaccgcagctcgattcgaacagctggtggtacttctagatggatactactatttagatatttgatatatttattttgcaacttgtttaatcagctcatttcgctttcgaaatgaattgggaggataagcttagcgtggcccacggctttgggccgcagaaattaattggagacgttggctcatctcatctctagggccgcacctacgtggtgcaacttgcgcagccacgatcgaatcgttcgagcgtgaaacccattgccgtcaccacctcgcctcatccctttcagggaccaatcggtttttagccctacgcgcccctgcgatcgcgacgcccacgatagctaaatcccgaaagcaaataagcagtaatcggacagcgactcgaccgggattagttaaacaatggcttgattaattagatgctggaatttggagccttctgataagtttagggcctgtttggcacagctccagctccagcttcaccccttctggagctggagctcagccaaacagtttcggctccaccaaaacggggagtggagctgggtggagctctctcacaaaatgaactagagttgtggagttgggtttaggcagctccacaactccactccagactcaactcctggagttaaatttaggagttggagctgtaccaaacaggcccttagttttgcacttggtactttaatttttttttgagtgagtgtaaatttgtttctaaactttgtttatgaatttgttttgtattggtgcagttggatgattgggtgctgtgccggatttacaacaagaagggcgggctggagaagccgccggccgcggcggtggcggcggcggggatggtgagcagcggcggcggcgtccagaggaagccgatggtgggggtgaacgcggcggtgagctccccgccggagcagaagccggtggtggcggggccggcgttcccggacctggcggcgtactacgaccggccgtcggactcgatgccgcggctgcacgccgactcgagctgctcggagcaggtgctgtcgccggagttcgcgtgcgaggtgcagagccagcccaagatcagcgagtgggagcgcaccttcgccaccgtcgggcccatcaaccccgccgcctccatcctcgaccccgccggctccggcggcctcggcggcctcggcggcggcggcagcgaccccctcctccaggacatcctcatgtactggggcaagccattctagacgaccaaaaaaaaaaaaaaacaaccgcattggcagcaatggtgtcactgaacaccgtgcaggctagctagcttcatggccggtgaactttgactcaggcgagccgccggagttgactcaaagataattaaaagaagtgttttaagtggattggattggattagacagaggagatgaggactcgagaaaggcggcgatgagaccgtggttggggggaccctggcctggactgaacgacgacgaggcagcagcagaaagatggtgcaattgcatcgggtggcatgtcagtgtgtgtgtatagtggcatgtacatagtacatggtgattgattcggtatacagggggctagctttcctgtttctgtttcttcattggttaattattactcccattataaggtcttcttcagggttgctagcttaattaattaattaattagcccagtggttgaagtgtaagtcaaaattcatcaagtcagagactggaataatacaatacagtactg</dnaseqindica>
External Link(s)	NCBI Gene:Os01g0884300, RefSeq:Os01g0884300