Identification of differentially expressed genes between sorghum genotypes with contrasting nitrogen stress tolerance by genome-wide transcriptional profiling

BackgroundSorghum is an important cereal crop, which requires large quantities of nitrogen fertilizer for achieving commercial yields. Identification of the genes responsible for low-N tolerance in sorghum will facilitate understanding of the molecular mechanisms of low-N tolerance, and also facilitate the genetic improvement of sorghum through marker-assisted selection or gene transformation. In this study we compared the transcriptomes of root tissues from seven sorghum genotypes having differential response to low-N stress.ResultsIllumina RNA-sequencing detected several common differentially expressed genes (DEGs) between four low-N tolerant sorghum genotypes (San Chi San, China17, KS78 and high-NUE bulk) and three sensitive genotypes (CK60, BTx623 and low-NUE bulk). In sensitive genotypes, N-stress increased the abundance of DEG transcripts associated with stress responses including oxidative stress and stimuli were abundant. The tolerant genotypes adapt to N deficiency by producing greater root mass for efficient uptake of nutrients. In tolerant genotypes, higher abundance of transcripts related to high affinity nitrate transporters (NRT2.2, NRT2.3, NRT2.5, and NRT2.6) and lysine histidine transporter 1 (LHT1), may suggest an improved uptake efficiency of inorganic and organic forms of nitrogen. Higher abundance of SEC14 cytosolic factor family protein transcript in tolerant genotypes could lead to increased membrane stability and tolerance to N-stress.ConclusionsComparison of transcriptomes between N-stress tolerant and sensitive genotypes revealed several common DEG transcripts. Some of these DEGs were evaluated further by comparing the transcriptomes of genotypes grown under full N. The DEG transcripts showed higher expression in tolerant genotypes could be used for transgenic over-expression in sensitive genotypes of sorghum and related crops for increased tolerance to N-stress, which results in increased nitrogen use efficiency for sustainable agriculture.

[1]  R. C. Muchow,et al.  Genotypic variation for grain yield and grain nitrogen concentration among sorghum hybrids under different levels of nitrogen fertiliser and water supply , 1998 .

[2]  F. Daniel-Vedele,et al.  Arabidopsis Roots and Shoots Show Distinct Temporal Adaptation Patterns toward Nitrogen Starvation1[W] , 2011, Plant Physiology.

[3]  Richard A. Dixon,et al.  Activation Tagging Identifies a Conserved MYB Regulator of Phenylpropanoid Biosynthesis , 2000, Plant Cell.

[4]  Ming Yan,et al.  Multiple roles of nitrate transport accessory protein NAR2 in plants , 2011, Plant signaling & behavior.

[5]  Rongchen Wang,et al.  Microarray Analysis of the Nitrate Response in Arabidopsis Roots and Shoots Reveals over 1,000 Rapidly Responding Genes and New Linkages to Glucose, Trehalose-6-Phosphate, Iron, and Sulfate Metabolism1[w] , 2003, Plant Physiology.

[6]  F Peter Guengerich,et al.  Complex reactions catalyzed by cytochrome P450 enzymes. , 2007, Biochimica et biophysica acta.

[7]  T. Zhu,et al.  Transcriptome response to nitrogen starvation in rice , 2012, Journal of Biosciences.

[8]  David Botstein,et al.  GO: : TermFinder--open source software for accessing Gene Ontology information and finding significantly enriched Gene Ontology terms associated with a list of genes , 2004, Bioinform..

[9]  G. Coruzzi,et al.  THE MOLECULAR-GENETICS OF NITROGEN ASSIMILATION INTO AMINO ACIDS IN HIGHER PLANTS. , 1996, Annual review of plant physiology and plant molecular biology.

[10]  J. Maranville,et al.  Physiological adaptations for nitrogen use efficiency in sorghum† , 2004, Plant and Soil.

[11]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[12]  G. Coruzzi,et al.  Nitrate-responsive miR393/AFB3 regulatory module controls root system architecture in Arabidopsis thaliana , 2010, Proceedings of the National Academy of Sciences.

[13]  R. Finlay,et al.  Expression analysis of Clavata1-like and Nodulin21-like genes from Pinus sylvestris during ectomycorrhiza formation , 2011, Mycorrhiza.

[14]  B. Hirel,et al.  An approach to the genetics of nitrogen use efficiency in maize. , 2004, Journal of experimental botany.

[15]  N. Crawford,et al.  Molecular and physiological aspects of nitrate uptake in plants , 1998 .

[16]  Daniel J. Cosgrove,et al.  Loosening of plant cell walls by expansins , 2000, Nature.

[17]  Matthijs Tollenaar,et al.  N uptake, N partitioning, and photosynthetic N-use efficiency of an old and a new maize hybrid , 1994 .

[18]  N. Crawford,et al.  Nitrate: nutrient and signal for plant growth. , 1995, The Plant cell.

[19]  F. Moradi,et al.  Comparison of the Drought Stress Responses of Tolerant and Sensitive Wheat Cultivars during Grain Filling: Impact of Invertase Activity on Carbon Metabolism during Kernel Development , 2011 .

[20]  T. Zhu,et al.  Genome-wide analysis of Arabidopsis responsive transcriptome to nitrogen limitation and its regulation by the ubiquitin ligase gene NLA , 2007, Plant Molecular Biology.

[21]  A. Hanson,et al.  Choline monooxygenase, an unusual iron-sulfur enzyme catalyzing the first step of glycine betaine synthesis in plants: prosthetic group characterization and cDNA cloning. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[22]  Cole Trapnell,et al.  Ultrafast and memory-efficient alignment of short DNA sequences to the human genome , 2009, Genome Biology.

[23]  J. Zhao,et al.  A soybean β-expansin gene GmEXPB2 intrinsically involved in root system architecture responses to abiotic stresses. , 2011, The Plant journal : for cell and molecular biology.

[24]  B. Forde,et al.  An Arabidopsis MADS box gene that controls nutrient-induced changes in root architecture. , 1998, Science.

[25]  R. W. Davis,et al.  Nitrate reductase from squash: cDNA cloning and nitrate regulation. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[26]  Rajeev K. Varshney,et al.  Differentially expressed genes between drought-tolerant and drought-sensitive barley genotypes in response to drought stress during the reproductive stage , 2009, Journal of experimental botany.

[27]  P. Waggoner,et al.  Nitrogen fertilizer: retrospect and prospect. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[28]  D. R. Hoagland,et al.  The Water-Culture Method for Growing Plants Without Soil , 2018 .

[29]  Yubi Huang,et al.  Differential global gene expression changes in response to low nitrogen stress in two maize inbred lines with contrasting low nitrogen tolerance , 2011, Genes & Genomics.

[30]  B. Ney,et al.  The challenge of improving nitrogen use efficiency in crop plants: towards a more central role for genetic variability and quantitative genetics within integrated approaches. , 2007, Journal of experimental botany.

[31]  William R. Raun,et al.  Improving Nitrogen Use Efficiency for Cereal Production , 1999 .

[32]  Michael F. Thomashow,et al.  PLANT COLD ACCLIMATION: Freezing Tolerance Genes and Regulatory Mechanisms. , 1999, Annual review of plant physiology and plant molecular biology.

[33]  N. Murata,et al.  Glycinebetaine protects plants against abiotic stress: mechanisms and biotechnological applications. , 2011, Plant, cell & environment.

[34]  Rongchen Wang,et al.  Genomic Analysis of a Nutrient Response in Arabidopsis Reveals Diverse Expression Patterns and Novel Metabolic and Potential Regulatory Genes Induced by Nitrate , 2000, Plant Cell.

[35]  Root Nitrogen Acquisition and Assimilation , 2005 .

[36]  B. Nyström,et al.  A phosphatidylserine decarboxylase activity in root cells of oat (Avena sativa) is involved in altering membrane phospholipid composition during drought stress acclimation. , 2006, Plant physiology and biochemistry : PPB.

[37]  A. Good,et al.  Can less yield more? Is reducing nutrient input into the environment compatible with maintaining crop production? , 2004, Trends in plant science.

[38]  Klaus Palme,et al.  Auxin transport inhibitors block PIN1 cycling and vesicle trafficking , 2001, Nature.

[39]  G. Lemaire,et al.  Nitrogen uptake capacities of maize and sorghum crops in different nitrogen and water supply conditions , 1996 .

[40]  Tong Zhu,et al.  Global transcription profiling reveals differential responses to chronic nitrogen stress and putative nitrogen regulatory components in Arabidopsis , 2007, BMC Genomics.

[41]  W. Frommer,et al.  Arabidopsis LHT1 Is a High-Affinity Transporter for Cellular Amino Acid Uptake in Both Root Epidermis and Leaf Mesophyll[W] , 2006, The Plant Cell Online.

[42]  L. Williams,et al.  Amino acid carriers of Ricinus communis expressed during seedling development: molecular cloning and expression analysis of two putative amino acid transporters, RcAAP1 and RcAAP2 , 1998, Plant Molecular Biology.

[43]  B. Møller,et al.  Plant cytochromes P450: tools for pharmacology, plant protection and phytoremediation. , 2003, Current opinion in biotechnology.

[44]  W. Jackson,et al.  Analysis and Interpretation of Factors Which Contribute to Efficiency of Nitrogen Utilization1 , 1982 .

[45]  G. Edwards,et al.  Single-cell C(4) photosynthesis versus the dual-cell (Kranz) paradigm. , 2004, Annual review of plant biology.

[46]  Rong Zhou,et al.  Identification of genes associated with nitrogen-use efficiency by genome-wide transcriptional analysis of two soybean genotypes , 2011, BMC Genomics.

[47]  Mark Stitt,et al.  The interaction between elevated carbon dioxide and nitrogen nutrition: the physiological and molecular background , 1999 .

[48]  Xianghua Li,et al.  Expression Profiles of 10,422 Genes at Early Stage of Low Nitrogen Stress in Rice Assayed using a cDNA Microarray , 2006, Plant Molecular Biology.

[49]  Tony Hunter,et al.  The regulation of transcription by phosphorylation , 1992, Cell.

[50]  Mark Stitt,et al.  Genome-Wide Reprogramming of Primary and Secondary Metabolism, Protein Synthesis, Cellular Growth Processes, and the Regulatory Infrastructure of Arabidopsis in Response to Nitrogen1[w] , 2004, Plant Physiology.

[51]  R. Socolow Nitrogen management and the future of food: lessons from the management of energy and carbon. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[52]  B. Williams,et al.  Mapping and quantifying mammalian transcriptomes by RNA-Seq , 2008, Nature Methods.

[53]  P. Vitousek,et al.  Significant Acidification in Major Chinese Croplands , 2010, Science.

[54]  C. Liu,et al.  Overexpression of the phosphatidylinositol synthase gene (ZmPIS) conferring drought stress tolerance by altering membrane lipid composition and increasing ABA synthesis in maize. , 2013, Plant, cell & environment.

[55]  T. Hunter,et al.  The protein kinase family: conserved features and deduced phylogeny of the catalytic domains. , 1988, Science.

[56]  D. Bush,et al.  LHT1, A Lysine- and Histidine-Specific Amino Acid Transporter in Arabidopsis , 1997, Plant physiology.

[57]  K. Shinozaki,et al.  Receptor-like protein kinase 2 (RPK 2) is a novel factor controlling anther development in Arabidopsis thaliana. , 2007, The Plant journal : for cell and molecular biology.

[58]  N. Crawford,et al.  Molecular and Developmental Biology of Inorganic Nitrogen Nutrition , 2002, The arabidopsis book.

[59]  T. Zhu,et al.  Increased nitrogen-use efficiency in transgenic rice plants over-expressing a nitrogen-responsive early nodulin gene identified from rice expression profiling. , 2009, Plant, cell & environment.

[60]  M. Robinson,et al.  A scaling normalization method for differential expression analysis of RNA-seq data , 2010, Genome Biology.

[61]  K. Taulavuori,et al.  After-effects of drought-related winter stress in previous and current year stems of Vaccinium myrtillus L , 2007 .