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Microarray Analysis on Salt Stress Affected Genes in Tomato Seedlings

ABSTRACT: Large-scale gene expression affected by salt stress was analyzed with tomato seedlings (Lycoperson esculentum Mill cv. Money Maker) by a cDNA microarray (Tom1). The significantly differentially expressed genes (5% Benjamini-Hochberg false discovery rate) consisted of 1757 sequences in the analyzed tissues (cotyledons + shoot tip). Genes with over 2 fold difference were selected from the list and further categorized into different function and cellular processes. Tomato homologous genes for the chaperone proteins, antioxidant enzymes (catalase and peroxidase), and ion transporters (Na+-driven multidrug efflux pump, vacuolar ATPase, and others) were induced. The ACC oxidase and ethylene-responsive gene tomato homologs had higher transcript level after salt treatment. Multiple members with different expression patterns were identified for the bZIP, WRKY, and MADS-box transcription regulator. Different genes in the signal transduction pathway, such as the protein kinases (Shaggy kinase, mitogen-activated protein kinase, ethylene receptor neverripe, and others), protein phosphatases, calmodulin, G-protein, and the N- myristoyltransferase, were regulated by salt stress. Most of the protease and the inhibitor homologs were suppressed by salt stress. In addition, different isoforms of cytochrome P450, genes for polyamine biosynthesis (putrescine and proline) and detoxification compounds (glutathione and thioredoxin), several key enzyme genes in the metabolic pathways of carbohydrates, amino acids, and fatty acids, were also affected by salt treatment. This study has provided a set of candidate genes, especially those in the regulatory machinery that can be further investigated to define salt stress in tomato and other plant species. Keywords: treatment response

ORGANISM(S): Solanum lycopersicum

PROVIDER: GSE8883 | GEO | 2007/09/05

SECONDARY ACCESSION(S): PRJNA102261

REPOSITORIES: GEO

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Project description:Large-scale gene expression affected by salt stress was analyzed with tomato seedlings (Lycoperson esculentum Mill cv. Money Maker) by a cDNA microarray (Tom1). The significantly differentially expressed genes (5% Benjamini-Hochberg false discovery rate) consisted of 1757 sequences in the analyzed tissues (cotyledons + shoot tip). Genes with over 2 fold difference were selected from the list and further categorized into different function and cellular processes. Tomato homologous genes for the chaperone proteins, antioxidant enzymes (catalase and peroxidase), and ion transporters (Na+-driven multidrug efflux pump, vacuolar ATPase, and others) were induced. The ACC oxidase and ethylene-responsive gene tomato homologs had higher transcript level after salt treatment. Multiple members with different expression patterns were identified for the bZIP, WRKY, and MADS-box transcription regulator. Different genes in the signal transduction pathway, such as the protein kinases (Shaggy kinase, mitogen-activated protein kinase, ethylene receptor neverripe, and others), protein phosphatases, calmodulin, G-protein, and the N- myristoyltransferase, were regulated by salt stress. Most of the protease and the inhibitor homologs were suppressed by salt stress. In addition, different isoforms of cytochrome P450, genes for polyamine biosynthesis (putrescine and proline) and detoxification compounds (glutathione and thioredoxin), several key enzyme genes in the metabolic pathways of carbohydrates, amino acids, and fatty acids, were also affected by salt treatment. This study has provided a set of candidate genes, especially those in the regulatory machinery that can be further investigated to define salt stress in tomato and other plant species. Keywords: treatment response Effect of 75mM NaCl on gene’s expression in tomato seedlings, Lycoperscon esculentum Mill. cv. Money Maker after 17 d of treatment. Each sample represents a pool of same tissue 20 seedlings grown in two flasks. Experiment was done in triplicate.

Project description:Purpose: Grafting is a commonly used cultural practice to counteract salt stress and is especially important for vegetable production. However, it is not clear which metabolic processes and genes are involved in the response of tomato rootstocks to salt stress. Our goals is to elucidate the regulatory mechanism through which grafting enhances salt tolerance. Methods: The salt-sensitive tomato variety Zhongza 9 and the strongly salt-tolerant tomato rootstock variety QZ-006 (selected by the research group in the early stage) were used as experimental materials and were purchased from the Vegetable and Flower Institute of the Chinese Academy of Agricultural Sciences (Beijing, CHN) and Beijing Kaixingelin Agricultural Technology Co., Ltd. (Beijing, CHN)；The leaves of the seedlings of every sample was added to 5 mL of HNO3 (65%~68%) after they were heated in a microwave digestion system for 2~3 hours；RNA-seq libraries were sequenced on an Illumina HiSeq X Ten platform；qRT‒PCR was performed according to the instructions of a Fast Super EvaGreen qPCR Master Mix Kit (US Everbright®, Inc.)；The amino acid contents in the tomato leaves were calculated by the external standard method. The chlorophyll of the seedlings was extracted with acetone-ethanol (1:1) ；An indirect enzyme-linked immunosorbent assay (ELISA) was used to determine auxin (IAA), gibberellin (GA), cytokinin (ZR), brassinolide (BR), abscisic acid (ABA), jasmonic acid (JA) and salicylic acid (SA) contents, and kit purchased from ThermoFisher Scientific Co., Ltd. (USA). Endogenous ethylene (ETH) was collected, fixed and determined according to the methods of Ling (Ling et al.,2008) using a Shimadzu 2010 gas chromatograph equipped with a hydrogen flame ion detector. ETH standard gas was purchased from Sigma Company. Results: Compared with the NGS, the GSs were more salt tolerant, and the Na+ content in the leaves decreased significantly. Through transcriptome sequencing data analysis of 36 samples, we found that GSs exhibited more stable gene expression patterns, with a lower number of DEGs. WRKY and PosF21 transcription factors were significantly upregulated in the GSs compared to the NGSs. Moreover, the GSs presented more amino acids, a higher photosynthetic index and a higher content of growth-promoting hormones. The main differences between GSs and NGSs were in the expression levels of genes involved in the BR signalling pathway, with significant upregulation of XTHs. Conclusions: Our study demonstrates that grafting on salt tolerant rootstocks can bring different metabolic processes and transcription levels changes to scion leaves, thereby the scion leaves show stronger salt tolerance. This information provides new insight into the mechanism underlying tolerance to salt stress regulation and provides useful molecular biological basis for improving plant salt resistance.

Project description:Gene expression patterns in roots of Camelina sativa with enhanced salinity tolerance arising from growth in soil treated with plant growth promoting bacteria producing 1-aminocyclopropane-1-carboxylate deaminase (ACC deaminase) or from expression of the corresponding acdS gene in transgenic lines. Salinity stress negatively affects crop production. However in camelina, grown in soils treated with PGPB producing 1-aminocyclopropane-1-carboxylate deaminase (acdS ) or transgenic lines expressing acdS exhibited increased salinity tolerance. AcdS reducing the level of stress ethylene to below the point where it is inhibitory to growth. Gene expression patterns in roots responding to salt stress was affected by the expression of acdS under the control of CaMV 35S or root-specific (rolD) promoters in transgenic lines, or by growth in soils treated with endophytic PGPB producing acdS indicate that the number of the genes were differentially expressed were more assigned to genome III in transgenic plants however in PGPB treated plants the number of the genes were differentially expressed were almost equally assigned to all three genomes. Different promoter may induce different set or even different homeologues genes in camelina with probably the same function in response to salt stress. Though root is not a photosynthetic tissue reduction of the ethylene in root cells has positive effect on plant photosynthetic machinery. The expression of the genes involved in minor CHO metabolism was up-regulated mainly in roots of acdS contain plants during salt stress. Moderate reduction in ethylene production has positive effect on root growth during salt stress but reduction of the ethylene higher than a certain level has negative effect on root growth due to reduction of the expression of the genes involved in root cell elongation. AcdS gene modulating the level of ROS in cells in the level that induce ROS signaling but preventing cellular damage by make a balance on up and down-regulation of the genes involved in oxidation-reduction process in root cells under salinity stress. The acdS containing PGPB (8R6) were mostly effected the ethylene signaling and ABA biosynthesis and signaling in positive way but transgenic line depends to the promoter affecting Auxin, JA and BR signaling or biosynthesis.

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Microarray Analysis on Salt Stress Affected Genes in Tomato Seedlings

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