Gene Profile Analysis and Molecular-Physiological Evaluation of Tomato Genotypes Under Drought Stress

Identification of the differentially-expressed genes is important for clarification of the complex molecular mechanisms under drought conditions. In this experiment, transcriptome profiles of sensitive and tolerant tomato genotypes under drought stress were analyzed. Three up-regulated genes were selected, included CAB3 (Chlorophyll a-b binding protein3), SAMDC (S-adenosylmethionine decarboxylase proenzyme), and ACS9 (1-aminocyclopropane-1-carboxylate synthase 9). After bioinformatics analysis, tomato genotypes were subjected to drought stress and gene expression was determined using Real-Time-PCR. Physiological parameters of genotypes were also measured by spectrophotometer-based methods. According to the results, these three genes play a key role in stress tolerance. Expression of the CAB3 gene in both sensitive and tolerant genotypes was not significantly different compared to the control, but the SAMDC gene decreased in both genotypes and the ACS9 gene decreased in sensitive genotype and increased in tolerant genotype. The physiological analysis also showed that under stress conditions, the photosynthetic system of the plant was disrupted and the chlorophyll content was reduced, but, proline content and antioxidant enzymes activity increased, in which their quantity in the tolerant genotype was significantly higher than sensitive. Under drought stress, due to damage to the lipid membrane, Malondialdehyde content also increased, in which the sensitive genotype was more affected.

Alteration in expression level of some growth and stress-related genes after rhizobacteria inoculation to alleviate drought tolerance in sensitive rice genotype

Background Drought stress is one of the major abiotic stresses that adversely affect rice production. Four rice genotypes, Giza177, IR64 (as sensitive genotypes) and Vandana, Orabi3 (as tolerant genotypes) were used to screen and characterize the soil microbes associated with each genotype under drought stress. Results The soil microbes associated with the tolerant genotypes showed high drought tolerance and high levels of enzyme activity. The most drought-tolerant isolates were inoculated with the sensitive genotype Giza177 under drought conditions. Some morphological, biochemical and molecular responses of inoculated plants were estimated. Inoculated plants showed regulation of some growth and stress-related genes (COX1, AP2-EREBP, GRAM, NRAMP6, NAM, GST, DHN and three genes of expansin (EXP1, EXP2 and EXP3) under drought conditions. Expression profiling of these genes were highly induced in plants inoculated with 4E11 and were correlated with improved growth status under drought stress. Conclusion Based on this, drought-tolerant plant growth-promoting rhizobacteria (PGPRs) were associated with the drought-tolerant genotype (Orabi 3). They were related to the significant increase in soil enzymes activities (dehydrogenase, nitrogenase, urease and alkaline phosphatase) in the rhizosphere of tolerant genotype. Inoculation the drought-sensitive genotype (Giza 177) with the most drought-tolerant isolates improved the tolerance status of the sensitive rice genotype and induced the expression of some growth and stress-responsive genes. AP2-EREBP, NRAMP6, DHN and all expansin genes (EXP1, EXP2 and EXP3) were the highly induced genes in inoculated plants with 4E11 strain and the consortium of three selected strains under drought condition. Graphic abstract

Comparative Physiological and Transcriptomic Analyses Reveal Ascorbate and Glutathione Coregulation of Cadmium Toxicity Resistance in Wheat Genotypes

Background: Understanding the cadmium (Cd) resistance mechanism is crucial for combating the phytotoxicity of Cd and meeting the increasing food demand daily. A classic symptom of Cd toxicity is root growth inhibition. Results: Using two wheat genotypes (Cd tolerant genotype T207 and Cd sensitive genotype S276) with differing root growths in response to Cd, we conducted comparative physiological and transcriptomic analyses and exogenous application tests to interpret Cd detoxification mechanisms. S276 accumulated more H2O2, O2-, and malonaldehyde than T207. Catalase activity and levels of ascorbic acid (AsA) and glutathione (GSH) were higher, whereas superoxide dismutase and peroxidase activities were lower in T207 than in S276. Transcriptome analysis showed that the expression of RBOHA, RBOHC, and RBOHE significantly increased, whereas that of RBOHB markedly decreased by Cd treatment. The transcriptional levels of 22 genes encoding RBOH were higher, and that of 11 genes were lower in T207 than in S276. The transcription of genes involved in the AsA-GSH cycle was profoundly reshaped by Cd treatment; 124 genes were higher and 43 genes were lower in T207 than in S276. Exogenous combined application of AsA and GSH alleviated Cd toxicity by scavenging excess ROS and coordinately modulating root length and branching, especially in S276.Conclusions: These results indicate that the AsA-GSH cycle fundamentally and vigorously influences plant defense against Cd toxicity, which provides valuable information for further clarification of the mechanisms underlying Cd detoxification.

Genome-wide analysis of long non-coding RNAs (lncRNAs) in two contrasting soybean genotypes subjected to phosphate starvation

Background Phosphorus (P) is essential for plant growth and development, and low-phosphorus (LP) stress is a major factor limiting the growth and yield of soybean. Long noncoding RNAs (lncRNAs) have recently been reported to be key regulators in the responses of plants to stress conditions, but the mechanism through which LP stress mediates the biogenesis of lncRNAs in soybean remains unclear. Results In this study, to explore the response mechanisms of lncRNAs to LP stress, we used the roots of two representative soybean genotypes that present opposite responses to P deficiency, namely, a P-sensitive genotype (Bogao) and a P-tolerant genotype (NN94156), for the construction of RNA sequencing (RNA-seq) libraries. In total, 4,166 novel lncRNAs, including 525 differentially expressed (DE) lncRNAs, were identified from the two genotypes at different P levels. GO and KEGG analyses indicated that numerous DE lncRNAs might be involved in diverse biological processes related to phosphate, such as lipid metabolic processes, catalytic activity, cell membrane formation, signal transduction, and nitrogen fixation. Moreover, lncRNA-mRNA-miRNA and lncRNA-mRNA networks were constructed, and the results identified several promising lncRNAs that might be highly valuable for further analysis of the mechanism underlying the response of soybean to LP stress. Conclusions These results revealed that LP stress can significantly alter the genome-wide profiles of lncRNAs, particularly those of the P-sensitive genotype Bogao. Our findings increase the understanding of and provide new insights into the function of lncRNAs in the responses of soybean to P stress.

Comparative Metabolomic and Transcriptomic Studies Reveal Key Metabolism Pathways Contributing to Freezing Tolerance Under Cold Stress in Kiwifruit

Cold stress poses a serious treat to cultivated kiwifruit since this plant generally has a weak ability to tolerate freezing tolerance temperatures. Surprisingly, however, the underlying mechanism of kiwifruit’s freezing tolerance remains largely unexplored and unknown, especially regarding the key pathways involved in conferring this key tolerance trait. Here, we studied the metabolome and transcriptome profiles of the freezing-tolerant genotype KL (Actinidia arguta) and freezing-sensitive genotype RB (A. arguta), to identify the main pathways and important metabolites related to their freezing tolerance. A total of 565 metabolites were detected by a wide-targeting metabolomics method. Under (−25°C) cold stress, KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway annotations showed that the flavonoid metabolic pathways were specifically upregulated in KL, which increased its ability to scavenge for reactive oxygen species (ROS). The transcriptome changes identified in KL were accompanied by the specific upregulation of a codeinone reductase gene, a chalcone isomerase gene, and an anthocyanin 5-aromatic acyltransferase gene. Nucleotides metabolism and phenolic acids metabolism pathways were specifically upregulated in RB, which indicated that RB had a higher energy metabolism and weaker dormancy ability. Since the LPCs (LysoPC), LPEs (LysoPE) and free fatty acids were accumulated simultaneously in both genotypes, these could serve as biomarkers of cold-induced frost damages. These key metabolism components evidently participated in the regulation of freezing tolerance of both kiwifruit genotypes. In conclusion, the results of this study demonstrated the inherent differences in the composition and activity of metabolites between KL and RB under cold stress conditions.

Light spectrum modifies the drought-induced changes of glutathione and free amino acid levels in wheat

The proposed spectral control of the adaptive metabolic responses to drought was tested by the comparison of two wheat genotypes with different stress tolerance. White light was used as the reference to see the effects of the three altered ratios of blue, red, and far-red spectral components, which conditions are referred to as blue, pink, and far-red lights. Under all spectral conditions, drought greatly reduced the growth of both genotypes. However, the glutathione content decreased and the proline level increased, independently of the spectrum, only in the sensitive genotype. Far-red light greatly decreased the amount of cystine and glutathione disulphide during the stress which resulted in their lower ratios compared to the reduced forms only in the tolerant genotype. The maintained more reducing redox environment contributes to its better stress tolerance. In far-red light, drought induced a greater accumulation of several free amino acids (mHis, Val, Ile, Leu, Asn, His, Tyr, Lys, Arg) in the tolerant genotype, while a smaller one in the sensitive genotype compared to the other spectral conditions. The transcript level of the genes related to amino acid and glutathione metabolism was also different between the two genotypes under this condition. The present results indicate the adaptive adjustment of glutathione and amino acid levels by far-red light during drought which observation can serve as a basis for the spectrum-dependent modification of the protective metabolites (glutathione, proline) of crops to reduce the stress-induced damages.

Involvement of Polyamine Metabolism in the Response of Medicago truncatula Genotypes to Salt Stress

Salinity constitutes one of the most important causes leading to severe reduction in plant yield. Several reports correlate the accumulation of polyamines in plants with tolerance to abiotic stress cues. The present study examined three Medicago truncatula genotypes with differing sensitivities to salinity (TN1.11, tolerant; Jemalong A17, moderately sensitive; TN6.18, sensitive), with the aim of examining the genotype-specific involvement of the polyamine metabolic pathway in plant response to salinity. The study was carried out with leaves harvested 48 h after watering plants with 200 mM NaCl. A comprehensive profile of free polyamines was determined using high performance liquid chromatography. All genotypes showed spermidine and spermine as the most abundant polyamines under control conditions. In salinity conditions, spermine levels increased at the expense of putrescine and spermidine, indicating a drift of polyamine metabolism towards the synthesis of increasing polycationic forms as a stress response. The increasing balance between high and low polycationic forms was clearly diminished in the salt-sensitive genotype TN6.18, showing a clear correlation with its sensitive phenotype. The polyamine metabolic profile was then supported by molecular evidence through the examination of polyamine metabolism transcript levels by RT-qPCR. General suppression of genes that are involved upstream in the PA biosynthetic pathway was determined. Contrarily, an induction in the expression of genes involved in the biosynthesis of spermine and spermidine was observed, in agreement with the metabolic analysis. A significant induction in diamino oxidase expression, involved in the catabolism of putrescine, was specifically found in the sensitive genotype ΤΝ6.18, indicating a distinct metabolic response to stress. Present findings highlight the involvement of polyamines in the defense response of Medicago genotypes showing sensitivity to salt stress.

Genome-wide analysis of long non-coding RNAs (lncRNAs) in two contrasting soybean genotypes subjected to phosphate starvation

Background: Phosphorus (P) is essential for plant growth and development, and low-phosphorus (LP) stress is a major factor limiting the growth and yield of soybean. Long noncoding RNAs (lncRNAs) have recently been reported to be key regulators in the responses of plants to stress conditions, but the mechanism through which LP stress mediates the biogenesis of lncRNAs in soybean remains unclear.Results: In this study, to explore the response mechanisms of lncRNAs to LP stress, we used the roots of two representative soybean genotypes that present opposite responses to P deficiency, namely, a P-sensitive genotype (Bogao) and a P-tolerant genotype (NN94156), for the construction of RNA sequencing (RNA-seq) libraries. In total, 4,166 novel lncRNAs, including 525 differentially expressed (DE) lncRNAs, were identified from the two genotypes at different P levels. GO and KEGG analyses indicated that numerous DE lncRNAs might be involved in diverse biological processes related to phosphate, such as lipid metabolic processes, catalytic activity, cell membrane formation, signal transduction, and nitrogen fixation. Moreover, lncRNA-mRNA-miRNA and lncRNA-mRNA networks were constructed, and the results identified several promising lncRNAs that might be highly valuable for further analysis of the mechanism underlying the response of soybean to LP stress.Conclusions: These results revealed that LP stress can significantly alter the genome-wide profiles of lncRNAs, particularly those of the P-sensitive genotype Bogao. Our findings increase the understanding of and provide new insights into the function of lncRNAs in the responses of soybean to P stress.

Investigation of biochemical changes in barley inoculated with Trichoderma harzianum T-22 under salt stress

Increases in soil salinity impacts growth and yield of agricultural plants by inhibiting plant functions. Soil salinization is increasing because of the pressure of a growing population on food supply. Genetically modified crops and plant breeding techniques are being used to produce plants tolerant to salt stress. However, interactions of fungal endophytes with crop plants can also improve tolerance and is a less expensive approach. Here, the role of Trichoderma harzianum T-22 in alleviating NaCl-induced stress in two barley genotypes (cv. Vlamingh and cv. Gairdner) has been investigated. Metabolomics using GC-MS for polar metabolites and LC-MS for lipids was employed to provide insights into the biochemical changes in barley roots inoculated with fungus during the early stages of interaction. T. harzianum increased the root length of both genotypes under controlled and saline conditions. The fungus reduced the relative concentration of sugars in both genotypes and caused no change in organic acids under saline conditions. Amino acids decreased only in cv. Gairdner in fungus-inoculated roots under saline conditions. Lipid analyses suggest that salt stress causes large changes in the lipid profile of roots but that inoculation with fungus greatly reduces the extent of these changes. By studying a tolerant and a sensitive genotype and their responses to salt and inoculation we have been able to develop hypotheses about what lipid species and metabolites may be involved in the tolerant genotype for its tolerance to salt and how fungal inoculation changes the response of the sensitive genotype to improve its tolerance.