The Effects of Heat Treatment on the Gene Expression of Several Heat Shock Protein Genes in Two Cultivars of Strawberry (original) (raw)
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TURKISH JOURNAL OF AGRICULTURE AND FORESTRY, 2016
The mechanism of tolerance to high temperatures was investigated in two strawberry (Fragaria × ananassa Duch) cultivars, 'Redlands Hope' ('R. Hope' , heat tolerant) and 'Cal. Giant 3' ('CG3' , heat sensitive). Leaves were collected from plants that were exposed to gradual heat stress and heat-shock stress separately. The contents of nonenzymatic antioxidants such as ascorbic acid (AsA) and glutathione (GSH) and the activities of enzymatic antioxidants such as ascorbate peroxidase (APX) (EC 1.11.1.11), catalase (CAT) (EC 1.11.1.6), and glutathione reductase (GR) (EC. 1.6.4.2) were measured followed by heat treatments. Additionally, proline content was determined, and heat shock proteins (HSPs) were analyzed with an immunoblotting method to investigate protein markers involved in the heat-stress tolerance of strawberry plants. The contents of AsA and GSH did not change depending on heat stress type, temperatures, or cultivars. While APX and CAT activities increased with high temperatures, GR activity was almost unchanged. The proline content of the cultivars increased in both treatments. Anti-HSP60 immunoblots revealed that a 23 kDa polypeptide was detected during the heat acclimation of strawberry cultivars. The intensity of the heat shock protein in 'R. Hope' plants was more than in 'CG3' plants. Thus, the accumulation of 23 kDa heat shock protein was correlated with the heat tolerance of the cultivars. In conclusion, strawberry leaf tissues of 'R. Hope' were found to enhance the structural stability of cellular membranes under high temperature by increasing both the activity of such enzymes as CAT and APX to activate the antioxidative systems and the expression of 23 kDa HSP.
BMC Plant Biology, 2014
Background: Temperature extremes represent an important limiting factor to plant growth and productivity. The present study evaluated the effect of hydroponic pretreatment of strawberry (Fragaria x ananassa cv. 'Camarosa') roots with an H 2 S donor, sodium hydrosulfide (NaHS; 100 μM for 48 h), on the response of plants to acute heat shock treatment (42°C, 8 h). Results: Heat stress-induced phenotypic damage was ameliorated in NaHS-pretreated plants, which managed to preserve higher maximum photochemical PSII quantum yields than stressed plants. Apparent mitigating effects of H 2 S pretreatment were registered regarding oxidative and nitrosative secondary stress, since malondialdehyde (MDA), H 2 O 2 and nitric oxide (NO) were quantified in lower amounts than in heat-stressed plants. In addition, NaHS pretreatment preserved ascorbate/glutathione homeostasis, as evidenced by lower ASC and GSH pool redox disturbances and enhanced transcription of ASC (GDH) and GSH biosynthetic enzymes (GS, GCS), 8 h after heat stress imposition. Furthermore, NaHS root pretreatment resulted in induction of gene expression levels of an array of protective molecules, such as enzymatic antioxidants (cAPX, CAT, MnSOD, GR), heat shock proteins (HSP70, HSP80, HSP90) and aquaporins (PIP). Conclusion: Overall, we propose that H 2 S root pretreatment activates a coordinated network of heat shock defense-related pathways at a transcriptional level and systemically protects strawberry plants from heat shock-induced damage.
Heat treatments and expansin gene expression in strawberry fruit
Scientia Horticulturae, 2011
Heat treatments have been applied in fruit postharvest technology for insect disinfestations, decay control, ripening delay and modification of fruit responses to other stresses. Heat treatment affects several aspects of fruit ripening, such as ethylene production and cell wall degradation probably through changes in gene expression and protein synthesis. In this paper, strawberries (Fragaria × ananassa Duch., cv Camarosa) at 50-75% red stage of ripening were heat-treated at 45 • C during 3 h in an air oven and then stored at 20 • C for 0, 4, 18, 24 and 48 h. Fruit firmness was determined and the expression of a set of expansin genes (FaEXP1, FaEXP2, FaEXP4, FaEXP5 and FaEXP6) was analyzed. The firmness of treated fruit was higher than that of control fruit 24 h after treatment, though the differences disappeared after 48 h at 20 • C. The analysis by northern-blot indicated that heat treatments affected differently the expression of expansin genes. The expression of FaEXP1, FaEXP2 and FaEXP6 was lower in treated fruit during the following 24 h post-treatment. The lower expression of these expansin genes could contribute to delay softening after heat treatment.
(265) HSP101 in the Model Strawberry Fragaria vesca
HortScience
Our lab has initiated a project to determine if specific proteins expressed by strawberry function as part of the thermotolerance system. We have developed tools for investigating the role of heat shock proteins (HSPs) and other gene products in strawberry thermotolerance. These tools include an inbred diploid testing system, EST sequence data, and molecular markers for heat tolerance. We developed an inbred line, 5AF7, of the diploid strawberry Fragariavesca for testing gene function because the diploid genome is small (164Mbp), the life cycle of the plant is short (about 4 months), the plant size is small (a plant will produce fruit in a 4-inch pot), some genetic work is already done, F. vesca is transformable with Agrobacteriumtumefaciens, and results should be transferable to the commercial octoploid varieties. A cDNA library was constructed in the pCMVsport 6.1 vector using combined RNA from batches of aseptically grown F. vesca seedlings treated to various elevated temperature...
Heat shock proteins (Hsps) and transcription factors (Hsfs) are considered as an important class of genes involved in plant’s response to heat stress. To elucidate the genotypic differences in rice in response to high temperature stress, plants were exposed to different levels of temperature ranging from 37 °C to 48 °C. The expression of genes belonging to Hsps and Hsf category were analyzed initially by digital microarray and later by semi-quantitative RT-PCR analysis using leaf tissues. Significant variations in the level of expression, timing of gene activation and maximum transcript accumulation were recorded among the set of rice genotypes selected based on their origin and phenotypic data recorded for heat stress. Genes encoding small Hsps (OsHsp 16, OsHsp 17.7 and OsHsp 18), OsHsp 70 DnaK, OsHsp 100 and OsHsf A2a showed strong induction upon heat stress with varied pattern and degrees. Out of six rice genotypes, two R-1389-RF-42 and Nagina22 (check heat tolerant) showed higher gene expression levels for most of the Hsps and Hsf genes tested with prominently better way of regulation which contributed to their greater heat tolerance and surmount the stress. In silico promoter analysis showed that strongly induced genes contain upstream regulatory elements corresponding to different stresses including heat shock, a good correlation was noted between in silico profiling of elements and their corresponding expression pattern. Information of such genotypic variation in expression levels of important candidate genes under heat stress supplemented with related field performance data could potentially be exploited in breeding programs for thermal stress tolerance. Keywords Abiotic stress . Differential expression . Heat shock proteins . Heat stress . Heat shock factors . In silico . Transcript Abbreviations HSR Heat shock response Hsf Heat stress transcription factors Hsp Heat shock proteins RT-PCR Reverse transcription PCR CT Canopy temperature
Functional & Integrative Genomics, 2014
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Role of Heat Shock Proteins in Improving Heat Stress Tolerance in Crop Plants
Heat Shock Proteins, 2016
High temperature response (HTR) or heat stress response (HSR) is a highly conserved phenomenon, which involves complex networks among different crop species. Heat stress usually results in protein dysfunction by improper folding of its linear amino acid chains to non-native proteins. This leads to unfavourable interactions and subsequent protein aggregation. To tackle this, plants have developed molecular chaperone machinery to maintain high quality proteins in the cell. This is governed by increasing the level of pre-existing molecular chaperones and by expressing additional chaperones through signalling mechanism. Dissecting the molecular mechanism by which plants counter heat stress and identifi cation of important molecules involved are of high priority. This could help in the development of plants with improved heat stress tolerance through advanced genomics and genetic engineering approaches. Owing to this reason molecular chaperones/Heat shock proteins (Hsps) are considered as potential candidates to address the issue of heat stress. In this chapter, recent progress on systematic analyses of heat shock proteins, their classifi cation and role in plant response to heat stress along with an overview of genomic and transgenic approaches to overcome the issue, are summarized.
Regulatory motifs found in the small heat shock protein (sHSP) gene family in tomato
BMC Genomics, 2018
Background: In living organisms, small heat shock proteins (sHSPs) are triggered in response to stress situations. This family of proteins is large in plants and, in the case of tomato (Solanum lycopersicum), 33 genes have been identified, most of them related to heat stress response and to the ripening process. Transcriptomic and proteomic studies have revealed complex patterns of expression for these genes. In this work, we investigate the coregulation of these genes by performing a computational analysis of their promoter architecture to find regulatory motifs known as heat shock elements (HSEs). We leverage the presence of sHSP members that originated from tandem duplication events and analyze the promoter architecture diversity of the whole sHSP family, focusing on the identification of HSEs. Results: We performed a search for conserved genomic sequences in the promoter regions of the sHSPs of tomato, plus several other proteins (mainly HSPs) that are functionally related to heat stress situations or to ripening. Several computational analyses were performed to build multiple sequence motifs and identify transcription factor binding sites (TFBS) homologous to HSF1AE and HSF21 in Arabidopsis. We also investigated the expression and interaction of these proteins under two heat stress situations in whole tomato plants and in protoplast cells, both in the presence and in the absence of heat shock transcription factor A2 (HsfA2). The results of these analyses indicate that different sHSPs are up-regulated depending on the activation or repression of HsfA2, a key regulator of HSPs. Further, the analysis of protein-protein interaction between the sHSP protein family and other heat shock response proteins (Hsp70, Hsp90 and MBF1c) suggests that several sHSPs are mediating alternative stress response through a regulatory subnetwork that is not dependent on HsfA2. Conclusions: Overall, this study identifies two regulatory motifs (HSF1AE and HSF21) associated with the sHSP family in tomato which are considered genomic HSEs. The study also suggests that, despite the apparent redundancy of these proteins, which has been linked to gene duplication, tomato sHSPs showed different up-regulation and different interaction patterns when analyzed under different stress situations.