Transcriptome analysis reveals differential gene expression in tomato under high-temperature stress

Tomato (Solanum lycopersicum) belongs to family Solanaceae that comprises more than 3000 species. Several studies are reported on various aspects of tomato genome. However, many important molecular processes have still not been studied in this important plant. Effect of fungal stress on differential gene expression was studied with the help of arbitrary primers. Differential gene expression under fungal infection showed expression of many genes that may have important implications; the expressed genes have been categorized in three categories viz. genes related to cellular metabolism, genes corresponding to defense related proteins and the genes related to poly(A) factors. Outcome of the project is expected to have important implications for the control of gene expression in tomato to improve yields and varieties.

Microrna Regulation of Nodule Zone-Specific Gene Expression In Soybean

High-temperature (HT) stress at flowering stage causes significant damage to soybean, including pollen abortion and fertilization failure, but few genes involved in male fertility regulation under HT stress in soybean have been characterized. Here, we demonstrated that miR156b-GmSPL2b module involved in male fertility regulation of soybean cytoplasmic male sterility (CMS)-based restorer line under HT stress. Overexpression of miR156b decreased male fertility in soybean CMS-based restorer line and its hybrid F1 with CMS line under HT stress. RNA-seq analysis found that miR156b mediated male fertility regulation in soybean under HT stress by regulating the expression of pollen development and HT response related genes. Metabolomic analysis of miR156bOE revealed reduction in flavonoid content under HT stress. Integrated transcriptomic and metabolomic analysis showed that the overexpression of miR156b caused flavonoid metabolism disorder in soybean flower bud under HT stress. Knockout of GmSPL2b also decreased the thermotolerance of soybean CMS-based restorer line during flowering. Moreover, GmSPL2b turned out to be directly bounded to the promoter of GmHSFA6b. Further verification indicated that GmHSFA6b overexpression enhanced HT tolerance in Arabidopsis during flowering. Substance content and gene expression analysis revealed that miR156b-GmSPL2b may mediate reactive oxygen species clearance by regulating flavonoid metabolism, thus participating in the regulation of male fertility in soybean under HT stress. This study not only provided important progress for understanding the molecular mechanism of miR156b-GmSPL2b regulating the male fertility of soybean CMS-based restorer line under HT stress, but also provided genetic resources and theoretical basis for creating HT-tolerant strong restorer lines.

During stress conditions, plant gene expression changes to cope up with the altered environment. The reprogramming of gene expression during viral infections is to be accounted towards host adaptation to the invading virus by developing defense mechanism. As a consequence of restructured gene expression, plant’s physiology gets affected and is evident in the form of symptoms. In the present study, Tomato leaf curl New Delhi virus (ToLCNDV) has been used as model system for the study of altered expression of genes in tomato during symptom development using microarray. Gene expression profiles were monitored in leaf tissues during symptom development in Solanum lycopersicum. To explore the differential expression analysis, tomato cDNA gene chip arrays have been used with 26,173 protein-coding genes. A total of 920 differentially expressed genes in response to ToLCNDV were identified in tomato. KEGG pathway analysis of these genes categorizes 652 differentially expressed genes into 77 known pathways, mostly related to increasing respiration rate, decreasing rate of photosynthesis, accumulation of soluble sugars/starch and results in elevated levels of amino acid synthesis. The possible role of these changes in altering host physiology, developed in the form of symptoms has been discussed.

A small heat shock protein GmHSP18.5a improves the male fertility restorability of cytoplasmic male sterility-based restorer line under high temperature stress in soybean

Differential activity and expression of antioxidant enzymes and alteration in osmolyte accumulation under high temperature stress in wheat seedlings

Crop productivity around the world is being seriously affected by adverse environmental conditions. High temperature (HT) stress has severely hampered plant growth, yield, and quality. Water spinach is a significant heat-resilient green leafy vegetable that can mitigate prolonged HT stress. However, the morphological, physiological, and biochemical alterations that occur in its response to heat stress remain unknown. In this study, the physiological response to HT stress in water spinach plants with different temperature (25-control, 30, 35, 40, 45 °C) tolerances was investigated. When plants were subjected to HT over a long period of time, their growth was stunted. The results showed that no significant difference was seen between the control (25 °C) and 30 °C for some traits (root shoot fresh weight, root morphological traits, and leaf gas exchanges parameters). Further, HT (35, 40, and 45 °C) stress significantly reduced the growth status, the gas exchange parameters, the pigment content, the photosystem function, and the root architecture system of water spinach. Conversely, HT stress considerably enhanced secondary metabolites in terms of total phenolics, flavonoids, soluble sugars, and anthocyanin content. Furthermore, heat stress remarkably increased the accumulation of reactive oxygen species (ROS) and caused cellular membrane damage. HT stress effectively altered the antioxidant defense system and caused oxidative damage. Generally, HT has an adverse effect on the enzyme activity of water spinach, leading to cell death. However, the current study found that temperatures ≥35 °C had an adverse effect on the growth of water spinach. Further research will be needed to examine the mechanism and the gene expression involved in the cell death that is caused by temperature stress in water spinach plants.

BackgroundPlant photoreceptors, phytochromes and cryptochromes, regulate many aspects of development and growth, such as seed germination, stem elongation, seedling de-etiolation, cotyledon opening, flower induction and circadian rhythms. There are several pieces of evidence of interaction between photoreceptors and phyto-hormones in all of these physiological processes, but little is known about molecular and genetic mechanisms underlying hormone-photoreceptor crosstalk.Methodology/Principal FindingsIn this work, we investigated the molecular effects of exogenous phyto-hormones to photoreceptor gene transcripts of tomato wt, as well as transgenic and mutant lines with altered cryptochromes, by monitoring day/night transcript oscillations. GA and auxin alter the diurnal expression level of different photoreceptor genes in tomato, especially in mutants that lack a working form of cryptochrome 1a: in those mutants the expression of some (IAA) or most (GA) photoreceptor genes is down regulated by these hormones.Conclusions/SignificanceOur results highlight the presence of molecular relationships among cryptochrome 1a protein, hormones, and photoreceptors' gene expression in tomato, suggesting that manipulation of cryptochromes could represent a good strategy to understand in greater depth the role of phyto-hormones in the plant photoperceptive mechanism.

Chemical and physical signals have been reported to mediate wound-induced proteinase inhibitor II (Pin2) gene expression in tomato and potato plants. Among the chemical signals, phytohormones such as abscisic acid (ABA) and jasmonic acid (JA) and the peptide systemin represent the best characterized systems. Furthermore, electrical and hydraulic mechanisms have also been postulated as putative Pin2-inducing systemic signals. Most of the chemical agents are able to induce Pin2 gene expression without any mechanical wounding. Thus, ABA, JA, and systemin initiate Pin2 mRNA accumulation in the directly treated leaves and in the nontreated leaves (systemic) that are located distal to the treated ones. ABA-deficient tomato and potato plants do not respond to wounding by accumulation of Pin2 mRNA, therefore providing a suitable model system for analysis of the signal transduction pathway involved in wound-induced gene activation. It was demonstrated that the site of action of JA is located downstream to the site of action of ABA. Moreover, systemin represents one of the initial steps in the signal transduction pathway regulating the wound response. Recently, it was reported that heat treatment and mechanical injury generate electrical signals, which propagate throughout the plant. These signals are capable of inducing Pin2 gene expression in the nontreated leaves of wounded plants. Furthermore, electrical current application to tomato leaves leads to an accumulation of Pin2 mRNA in local and systemic tissues. Examination of photosynthetic parameters (assimilation and transpiration rate) on several types of stimuli suggests that heat-induced Pin2 gene expression is regulated by an alternative pathway from that mediating the electrical current and mechanical wound response.

This study investigates the impact of IbACP (Ipomoea batatas anti-cancer peptide) on defense-related gene expression in tomato leaves, focusing on its role in plant defense mechanisms. Previously, IbACP was isolated from sweet potato leaves, and it was identified as a peptide capable of inducing an alkalinization response in tomato suspension culture media. Additionally, IbACP was found to regulate the proliferation of human pancreatic adenocarcinoma cells. Elucidate IbACP's molecular influence on defense-related gene expression in tomato leaves using next-generation sequencing analysis. To assess the impact of IbACP on defense-related gene expression, transcriptome data were analyzed, encompassing various functional categories such as photosynthesis, metabolic processes, and plant defense. Semi-quantitative reverse-transcription polymerase chain reaction analysis was employed to verify transcription levels of defense-related genes in tomato leaves treated with IbACP for durations ranging from 0 h (control) to 24 h. IbACP induced jasmonic acid-related genes (LoxD and AOS) at 2 h, with a significant up-regulation of salicylic acid-dependent gene NPR1 at 24 h. This suggested a temporal antagonistic effect between jasmonic acid and salicylic acid during the early hours of IbACP treatment. Downstream ethylene-responsive regulator genes (ACO1, ETR4, and ERF1) were consistently down-regulated by IbACP at all times. Additionally, IbACP significantly up-regulated the gene expressions of suberization-associated anionic peroxidases (TMP1 and TAP2) at all time points, indicating enhanced suberization of the plant cell wall to prevent pathogen invasion. IbACP enhances the synthesis of defense hormones and up-regulates downstream defense genes, improving the plant's resistance to biotic stresses.

Reference gene selection for quantitative real-time PCR normalization in tomato subjected to nitrogen, cold, and light stress

Three ionophores were used to investigate a potential role of the plasma-membrane (PM) potential in the regulation of systemic wound-response gene expression in tomato (Lycopersicon escuilentum Mill.) plants. Valinomycin, nigericin, and gramicidin, which affect the PM potential by dissipating H+ and K+ gradients, respectively, induced the rapid accumulation of wound-response gene transcripts. Transcript induction by gramicidin was kinetically, qualitatively and quantitatively similar to systemin-induced transcript accumulation. On a molar basis, gramicidin and nigericin, which affect gradients of both H+ and K+, were more effective than the K+-selective valinomycin. Hyperpolarization of the PM by fusicoccin, on the other hand, repressed wound-response gene expression and, at the same time, induced salicylic acid (SA) accumulation and the expression of pathogenesis-related proteins. We show here that the inhibition of the wound response after fusicoccin treatment is not mediated by elevated concentrations of SA but is likely a direct effect of PM hyperpolarization. The data indicate a role for the PM potential in the differential regulation of wound and pathogen defense responses.

The response of insects to stress, particularly starvation and high temperature stress, is a crucial area of insect research. Uridine diphosphate-glucosyltransferases (UGTs) are key enzymes involved in the detoxification of exogenous substances. This study analysed the role of the UGT344J7 gene in the response of Rhopalosiphum padi to starvation and high temperature stress. UGT344J7 was significantly upregulated under conditions of high temperature and food scarcity. Following RNAi targeting UGT344J7, the mortality of R. padi increased significantly under both high temperature and starvation conditions. Knockdown of the UGT344J7 gene led to a significant increase in reactive oxygen species (ROS) levels in R. padi, accompanied by a significant downregulation of four heat shock protein genes (Hsp70-1, Hsp70-2, Hsp68, Hsp90). Based on these results, we speculate that UGT344J7 regulates the expression of heat shock protein genes by modulating ROS levels, thereby helping R. padi cope with high temperature and starvation stress. This is the first report on the role of the UGT gene in starvation and high temperature stress in an aphid species. This research suggests that silencing UGT344J7 could serve as a potential strategy for controlling R. padi, and novel insecticides targeting this gene may be developed to disrupt the physiological processes of this significant pest.

The red imported fire ant (RIFA), Solenopsis invicta Buren is native to South America and is known as a global problematic invasive species. This study focused on the molecular response of RIFA by comparing gene expression profiles after exposing ants to low (10 °C) and high (40 °C) temperature stress and comparing them to untreated controls (30 °C). A total of 99,085 unigenes (the clustered non-redundant transcripts that are filtered from the longest assembled contigs) were obtained, of which 19,154 were annotated with gene descriptions, gene ontology terms, and metabolic pathways. 86 gene ontology (GO) functional sub-groups and 23 EggNOG terms resulted. Differentially expressed genes (DEGs) with log2FC ≥ 10 were screened and were compared at different temperatures. We found 203, 48, and 66 specific DEGs co-regulated at 10, 20, and 40 °C. Comparing transcriptome profiles for differential gene expression resulted in various DE genes, including cytochrome P450, NADH dehydrogenase subunit 1, cuticle protein and heat shock protein (HSP), which have previously been reported to be involved in cold and high temperature resistance. GO analysis revealed that antioxidant activity is up-regulated under high temperature stress. We verified the RNA-seq data by qPCR on 20 up- and down-regulated DEGs. These findings provide a basis for future understanding of the adaptation mechanisms of RIFA and the molecular mechanisms underlying the response to low and high temperatures.

Soil treatment-induced differential gene expression in tomato: Relationships between defense gene expression and soil microbial community composition