Molecular cloning, sequence characterization and tissue-specific expression of six NAC-like genes in soybean (Glycine max (L.) Merr.).

NAC proteins have been considered as one of the novel classes of plant-specific transcription factors functioning in diverse and vital physiological processes during plant development. In this study, six NAC-like genes from soybean, designated as GmNAC1-GmNAC6, were cloned and characterized. They each contained two introns and three exons and shared conservative structure of genomic organization. The predicted proteins, GmNAC1-GmNAC6, were similar in sequences, especially in NAC domain regions. However, RT-PCR analysis indicated that each GmNAC gene exhibited a specific expression pattern in tissues examined. GmNAC2, 3, 4, and 6 were expressed in most tested tissues while GmNAC1 and GmNAC5 were limited to a few tissues. In addition, expression patterns of GmNAC genes were characterized during seed filling and coordinated expression was observed between GmNAC genes. Finally, based on phylogenetic analysis, six GmNAC proteins were classed into five subgroups with different putative functions. To our knowledge, this is the first report on molecular cloning and initial characterization of NAC-like genes in soybean. Our results may provide the basis for future investigations of NAC-like genes' roles in seed development and other physiological processes in this important crop.

[1]  M. K. Jensen,et al.  Interactions between plant RING-H2 and plant-specific NAC (NAM/ATAF1/2/CUC2) proteins: RING-H2 molecular specificity and cellular localization. , 2003, The Biochemical journal.

[2]  J. Mol,et al.  The No Apical Meristem Gene of Petunia Is Required for Pattern Formation in Embryos and Flowers and Is Expressed at Meristem and Primordia Boundaries , 1996, Cell.

[3]  Bin Han,et al.  Gene Expression Phenotypes of Arabidopsis Associated with Sensitivity to Low Temperatures[w] , 2003, Plant Physiology.

[4]  E. Meyerowitz,et al.  A Homolog of NO APICAL MERISTEM Is an Immediate Target of the Floral Homeotic Genes APETALA3/PISTILLATA , 1998, Cell.

[5]  K. Hibara,et al.  The NAC domain mediates functional specificity of CUP-SHAPED COTYLEDON proteins. , 2004, The Plant journal : for cell and molecular biology.

[6]  Shoshi Kikuchi,et al.  Comprehensive analysis of NAC family genes in Oryza sativa and Arabidopsis thaliana. , 2003, DNA research : an international journal for rapid publication of reports on genes and genomes.

[7]  Takayuki Asano,et al.  Molecular characterization of ONAC300, a novel NAC gene specifically expressed at early stages in various developing tissues of rice , 2005, Molecular Genetics and Genomics.

[8]  H Fujisawa,et al.  Genes involved in organ separation in Arabidopsis: an analysis of the cup-shaped cotyledon mutant. , 1997, The Plant cell.

[9]  W. J. Lucas,et al.  Phloem long-distance transport of CmNACP mRNA: implications for supracellular regulation in plants. , 1999, Development.

[10]  A. Kinney,et al.  Genetic Modification Removes an Immunodominant Allergen from Soybean1,212 , 2003, Plant Physiology.

[11]  D. Hegedus,et al.  Molecular characterization of Brassicanapus NAC domain transcriptional activators induced in response to biotic and abiotic stress , 2003, Plant Molecular Biology.

[12]  Jianping Lu,et al.  CUPULIFORMIS establishes lateral organ boundaries in Antirrhinum , 2004, Development.

[13]  W. Fehr,et al.  Stage of Development Descriptions for Soybeans, Glycine Max (L.) Merrill , 1971 .

[14]  Thomas L. Madden,et al.  Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. , 1997, Nucleic acids research.

[15]  David P. Bartel,et al.  MicroRNAs: At the Root of Plant Development?1 , 2003, Plant Physiology.

[16]  K. Skriver,et al.  Structure of the conserved domain of ANAC, a member of the NAC family of transcription factors , 2004, EMBO reports.

[17]  Karam B. Singh,et al.  Transcription factors in plant defense and stress responses. , 2002, Current opinion in plant biology.

[18]  K. Hibara,et al.  The CUP-SHAPED COTYLEDON1 gene of Arabidopsis regulates shoot apical meristem formation. , 2001, Development.

[19]  Kazuo Shinozaki,et al.  A dehydration-induced NAC protein, RD26, is involved in a novel ABA-dependent stress-signaling pathway. , 2004, The Plant journal : for cell and molecular biology.

[20]  T. Vernoux,et al.  Roles of PIN-FORMED1 and MONOPTEROS in pattern formation of the apical region of the Arabidopsis embryo. , 2002, Development.

[21]  H. Furuhashi,et al.  A novel class of plant-specific zinc-dependent DNA-binding protein that binds to A/T-rich DNA sequences. , 2001, Nucleic acids research.

[22]  Diana V. Dugas,et al.  MicroRNA Regulation of NAC-Domain Targets Is Required for Proper Formation and Separation of Adjacent Embryonic, Vegetative, and Floral Organs , 2004, Current Biology.

[23]  D. Gingerich,et al.  Global and Hormone-Induced Gene Expression Changes during Shoot Development in Arabidopsis , 2002, The Plant Cell Online.

[24]  Roderic D. M. Page,et al.  TreeView: an application to display phylogenetic trees on personal computers , 1996, Comput. Appl. Biosci..

[25]  R. Shoemaker,et al.  Genome duplication in soybean (Glycine subgenus soja). , 1996, Genetics.

[26]  T. Boller,et al.  Differential induction of two potato genes, Stprx2 and StNAC, in response to infection by Phytophthora infestans and to wounding , 2001, Plant Molecular Biology.

[27]  M. Kozak Initiation of translation in prokaryotes and eukaryotes. , 1999, Gene.

[28]  M. Hajduch,et al.  A Systematic Proteomic Study of Seed Filling in Soybean. Establishment of High-Resolution Two-Dimensional Reference Maps, Expression Profiles, and an Interactive Proteome Database1[w] , 2005, Plant Physiology.

[29]  Addie Nina Olsen,et al.  NAC transcription factors: structurally distinct, functionally diverse. , 2005, Trends in plant science.

[30]  D. Grierson,et al.  Cloning and characterization of tomato leaf senescence-related cDNAs , 1997, Plant Molecular Biology.

[31]  D. Israel,et al.  Regulation of seed protein concentration in soybean by supra-optimal nitrogen supply , 2000 .

[32]  T. Boller,et al.  Purification of the trehalase GMTRE1 from soybean nodules and cloning of its cDNA. GMTRE1 is expressed at a low level in multiple tissues. , 1999, Plant physiology.

[33]  Kazuo Shinozaki,et al.  Isolation and Functional Analysis of Arabidopsis Stress-Inducible NAC Transcription Factors That Bind to a Drought-Responsive cis-Element in the early responsive to dehydration stress 1 Promoterw⃞ , 2004, The Plant Cell Online.

[34]  R. Gardner,et al.  Genomic fingerprinting by microsatellite-primed PCR: a critical evaluation. , 1995, PCR methods and applications.

[35]  N. Suzuki,et al.  A maize DNA-binding factor with a bZIP motif is induced by low temperature , 1995, Molecular and General Genetics MGG.

[36]  Diana V. Dugas,et al.  MicroRNA regulation of gene expression in plants. , 2004, Current opinion in plant biology.

[37]  C. Gutiérrez,et al.  GRAB proteins, novel members of the NAC domain family, isolated by their interaction with a geminivirus protein , 1999, Plant Molecular Biology.

[38]  J. Thompson,et al.  The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. , 1997, Nucleic acids research.

[39]  J. Randles,et al.  A NAC Domain Protein Interacts with Tomato leaf curl virus Replication Accessory Protein and Enhances Viral Replication , 2005, The Plant Cell Online.

[40]  J. Schell,et al.  Light‐induced expression of the chimeric chalcone synthase‐NPTII gene in tobacco cells , 1986, The EMBO journal.

[41]  N. Guex,et al.  SWISS‐MODEL and the Swiss‐Pdb Viewer: An environment for comparative protein modeling , 1997, Electrophoresis.

[42]  Tzung-Fu Hsieh,et al.  Molecular characterization of AtNAM: a member of theArabidopsis NAC domain superfamily , 2002, Plant Molecular Biology.

[43]  M. Ueguchi-Tanaka,et al.  Molecular analysis of the NAC gene family in rice , 2000, Molecular and General Genetics MGG.

[44]  F. Qu,et al.  HRT Gene Function Requires Interaction between a NAC Protein and Viral Capsid Protein to Confer Resistance to Turnip Crinkle Virus , 2000, Plant Cell.

[45]  D. Bartel MicroRNAs Genomics, Biogenesis, Mechanism, and Function , 2004, Cell.

[46]  N. Chua,et al.  MicroRNA Directs mRNA Cleavage of the Transcription Factor NAC1 to Downregulate Auxin Signals for Arabidopsis Lateral Root Development , 2005, The Plant Cell Online.

[47]  N. Chua,et al.  Arabidopsis NAC1 transduces auxin signal downstream of TIR1 to promote lateral root development. , 2000, Genes & development.

[48]  Jay J Thelen,et al.  High-throughput peptide mass fingerprinting of soybean seed proteins: automated workflow and utility of UniGene expressed sequence tag databases for protein identification. , 2004, Phytochemistry.