Stress-associated Protein Gene Research Articles

Influence of some abiotic factors on the apple stressassociated protein gene expression profiles

In plants, stress-associated proteins (SAP) play a crucial role in responding to adverse biotic and abiotic factors. In this study, we evaluated the expression profiles of 14 apple SAP genes (MdSAP) under the influence of low and high temperatures, as well as salinity, using the quantitative polymerase chain reaction (qPCR). The results showed that the most significant changes in expression levels were observed in the genes MdSAP11, MdSAP2, and MdSAP3 under high temperature conditions, MdSAP1, MdSAP2, MdSAP4, and MdSAP6 under low temperature conditions, and MdSAP1, MdSAP8, and MdSAP11 under salt stress. Furthermore, there was a tendency for gene expression to increase at 2 and/or 4 hours of exposure followed by a decrease at 24 hours. The analysis of the correlation between the expression levels of MdSAP genes revealed both positive and negative linear relationships.These findings will help elucidate the specific roles of individual SAP genes in shaping the apple stress response to low and high temperatures, as well as salinity.

5-Aminolevulinic acid treatment mitigates pesticide stress in bean seedlings by regulating stress-related gene expression and retrotransposon movements

Overdoses of pesticides lead to a decrease in the yield and quality of plants, such as beans. The unconscious use of deltamethrin, one of the synthetic insecticides, increases the amount of reactive oxygen species (ROS) by causing oxidative stress in plants. In this case, plants tolerate stress by activating the antioxidant defense mechanism and many genes. 5-Aminolevulinic acid (ALA) improves tolerance to stress by acting exogenously in low doses. There are many gene families that are effective in the regulation of this mechanism. In addition, one of the response mechanisms at the molecular level against environmental stressors in plants is retrotransposon movement. In this study, the expression levels of superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), glutathione reductase (GR), and stress-associated protein (SAP) genes were determined by Q-PCR in deltamethrin (0.5 ppm) and various doses (20, 40, and 80 mg/l) of ALA-treated bean seedlings. In addition, one of the response mechanisms at the molecular level against environmental stressors in plants is retrotransposon movement. It was determined that deltamethrin increased the expression of SOD (1.8-fold), GPX (1.4-fold), CAT (2.7-fold), and SAP (2.5-fold) genes, while 20 and 40 mg/l ALA gradually increased the expression of these genes at levels close to control, but 80 mg/l ALA increased the expression of these genes almost to the same level as deltamethrin (2.1-fold, 1.4-fold, 2.6-fold, and 2.6-fold in SOD, GPX, CAT, and SAP genes, respectively). In addition, retrotransposon-microsatellite amplified polymorphism (REMAP) was performed to determine the polymorphism caused by retrotransposon movements. While deltamethrin treatment has caused a decrease in genomic template stability (GTS) (27%), ALA treatments have prevented this decline. At doses of 20, 40, and 80 mg/L of ALA treatments, the GTS ratios were determined to be 96.8%, 74.6%, and 58.7%, respectively. Collectively, these findings demonstrated that ALA has the utility of alleviating pesticide stress effects on beans.

Genome-Wide Identification and Characterisation of Stress-Associated Protein Gene Family to Biotic and Abiotic Stresses of Grapevine.

Grapevine is one of the earliest domesticated fruit crops and prized for its table fruits and wine worldwide. However, the concurrence of a number of biotic/abiotic stresses affects their yield. Stress-associated proteins (SAPs) play important roles in response to both biotic and abiotic stresses in plants. Despite the growing number of studies on the genomic organisation of SAP gene family in various species, little is known about this family in grapevines (Vitis vinifera L.). In this study, a total of 15 genes encoding proteins possessing A20/AN1 zinc-finger were identified based on the analysis of several genomic and proteomic grapevine databases. According to their structural and phylogenetics features, the identified SAPs were classified into three main groups. Results from sequence alignments, phylogenetics, genomics structure and conserved domains indicated that grapevine SAPs are highly and structurally conserved. In order to shed light on their regulatory roles in growth and development, as well as the responses to biotic/abiotic stresses in grapevine, the expression profiles of SAPs were examined in publicly available microarray data. Bioinformatics analysis revealed distinct temporal and spatial expression patterns of SAPs in various tissues, organs and developmental stages, as well as in response to biotic/abiotic stresses. This study provides insight into the evolution of SAP genes in grapevine and may aid in efforts for further functional identification of A20/AN1-type proteins in the signalling cross-talking induced by biotic/abiotic stresses.

Identification and Analysis of Stress-Associated Proteins (SAPs) Protein Family and Drought Tolerance of ZmSAP8 in Transgenic Arabidopsis.

Stress-associated proteins (SAPs), a class of A20/AN1 zinc finger proteins, play vital roles in plant stress response. However, investigation of SAPs in maize has been very limited. Herein, to better trace the evolutionary history of SAPs in maize and plants, 415 SAPs were identified in 33 plant species and four species of other kingdoms. Moreover, gene duplication mode exploration showed whole genome duplication contributed largely to SAP gene expansion in angiosperms. Phylogeny reconstruction was performed with all identified SAPs by the maximum likelihood (ML) method and the SAPs were divided into five clades. SAPs within the same clades showed conserved domain composition. Focusing on maize, nine ZmSAPs were identified. Further promoter cis-elements and stress-induced expression pattern analysis of ZmSAPs indicated that ZmSAP8 was a promising candidate in response to drought stress, which was the only AN1-AN1-C2H2-C2H2 type SAP in maize and belonged to clade I. Additionally, ZmSAP8 was located in the nucleus and had no transactivation activity in yeast. Overexpressing ZmSAP8 enhanced the tolerance to drought stress in Arabidopsis thaliana, with higher seed germination and longer root length. Our results should benefit the further functional characterization of ZmSAPs.

Comprehensive Identification and Functional Analysis of Stress-Associated Protein (SAP) Genes in Osmotic Stress in Maize.

Stress-associated proteins (SAPs) are a kind of zinc finger protein with an A20/AN1 domain and contribute to plants' adaption to various abiotic and biological stimuli. However, little is known about the SAP genes in maize (Zea mays L.). In the present study, the SAP genes were identified from the maize genome. Subsequently, the protein properties, gene structure and duplication, chromosomal location, and cis-acting elements were analyzed by bioinformatic methods. Finally, their expression profiles under osmotic stresses, including drought and salinity, as well as ABA, and overexpression in Saccharomyces cerevisiae W303a cells, were performed to uncover the potential function. The results showed that a total of 10 SAP genes were identified and named ZmSAP1 to ZmSAP10 in maize, which was unevenly distributed on six of the ten maize chromosomes. The ZmSAP1, ZmSAP4, ZmSAP5, ZmSAP6, ZmSAP7, ZmSAP8 and ZmSAP10 had an A20 domain at N terminus and AN1 domain at C terminus, respectively. Only ZmSAP2 possessed a single AN1 domain at the N terminus. ZmSAP3 and ZmSAP9 both contained two AN1 domains without an A20 domain. Most ZmSAP genes lost introns and had abundant stress- and hormone-responsive cis-elements in their promoter region. The results of quantitative real-time PCR showed that all ZmSAP genes were regulated by drought and saline stresses, as well as ABA induction. Moreover, heterologous expression of ZmSAP2 and ZmSAP7 significantly improved the saline tolerance of yeast cells. The study provides insights into further underlying the function of ZmSAPs in regulating stress response in maize.

Genome-Wide Identification of the A20/AN1 Zinc Finger Protein Family Genes in Ipomoea batatas and Its Two Relatives and Function Analysis of IbSAP16 in Salinity Tolerance.

Stress-associated protein (SAP) genes—encoding A20/AN1 zinc-finger domain-containing proteins—play pivotal roles in regulating stress responses, growth, and development in plants. They are considered suitable candidates to improve abiotic stress tolerance in plants. However, the SAP gene family in sweetpotato (Ipomoea batatas) and its relatives is yet to be investigated. In this study, 20 SAPs in sweetpotato, and 23 and 26 SAPs in its wild diploid relatives Ipomoea triloba and Ipomoea trifida were identified. The chromosome locations, gene structures, protein physiological properties, conserved domains, and phylogenetic relationships of these SAPs were analyzed systematically. Binding motif analysis of IbSAPs indicated that hormone and stress responsive cis-acting elements were distributed in their promoters. RT-qPCR or RNA-seq data revealed that the expression patterns of IbSAP, ItbSAP, and ItfSAP genes varied in different organs and responded to salinity, drought, or ABA (abscisic acid) treatments differently. Moreover, we found that IbSAP16 driven by the 35 S promoter conferred salinity tolerance in transgenic Arabidopsis. These results provided a genome-wide characterization of SAP genes in sweetpotato and its two relatives and suggested that IbSAP16 is involved in salinity stress responses. Our research laid the groundwork for studying SAP-mediated stress response mechanisms in sweetpotato.

Genome-wide in silico identification and characterization of the stress associated protein (SAP) gene family encoding A20/AN1 zinc-finger proteins in potato (Solanum tuberosum L.).

Stress associated proteins (SAPs) in plants have a key role in providing tolerance to multiple abiotic stresses. SAP gene family in Solanum tuberosum has not been fully studied before. This study identified 17 StSAP genes in S. tuberosum which code for A20/AN1 zinc-finger proteins. All the genes were distributed on ten different chromosomes and six segmental duplication events were identified. The SAPs in S. tuberosum and its orthologs in Arabidopsis thaliana were classified into six groups through the phylogenetic analysis. Introns across StSAP genes were identified in four genes. The promotor study of the StSAP genes showed different hormone and stress-related cis-elements that could potentially have a role in environmental stress response. The expression of StSAP genes in response to heat, mannitol, and salt were analyzed through in silico transcriptomic analysis. This study could potentially help in further understanding the functions of SAP genes in S. tuberosum.

GmSAP5, a soybean A20/AN1 domain-containing stress-associated protein gene activated by GmAREB3, increases drought stress resistance in soybean by mediating ABA signaling

Drought stress impairs crop growth and productivity. Stress-associated proteins (SAPs), a class of zinc finger proteins containing the A20/AN1 domain, function in various stress responses in plants. However, little is known about the function of SAPs in drought-stress responses in soybean, an oil and protein crop. We report that a GmSAP5 protein confers drought tolerance by increasing sensitivity to abscisic acid (ABA) and reducing stomatal aperture. Overexpression and RNA interference of GmSAP5 in soybean hairy roots resulted in elevated resistance and sensitivity to drought stress, respectively. ABA and proline contents increased in GmSAP5-overexpressing plants under water-deficit conditions. Lower water loss rates and higher relative water contents were observed in GmSAP5-overexpressing plants, resulting in increased drought-stress resistance. A yeast one-hybrid assay and luciferase transient transcriptional activity assay showed that GmAREB3, an AREB/ABF transcription factor, could bind to the promoter of GmSAP5 and activate its expression. These results suggest that GmSAP5 acts downstream of GmAREB3 and improves drought-stress resistance by mediating ABA signaling.

Identification and Expression Analysis of Zinc Finger A20/AN1 Stress-Associated Genes SmSAP Responding to Abiotic Stress in Eggplant

Stress-associated proteins (SAP), a class of zinc-finger proteins, have been identified as novel stress regulatory proteins in stress responses. However, SAP genes in eggplant (SmSAP) have been little reported. It has important significance in identifying SAP members, understanding the molecular mechanisms underlying stress responses, and tolerance. We performed a comprehensive study of the A20/AN1 domains, motifs, gene structures, phylogenetic relationships, chromosomal locations, gene replications, collinearity, cis-acting elements, and expression pattern responses to various abiotic stresses. Twenty-one SAP genes were identified in eggplant (SmSAP) and were localized on 10 chromosomes. A phylogenetic analysis revealed that most of the SmSAP proteins showed a high homology with the tomato SAP members, and 21 members were divided into four groups based on the homology of the SAP members in eggplant, tomato, rice, and Arabidopsis. Further analysis revealed that SmSAP proteins contain the characteristic A20/AN1 domains, the A20 domain composed of motif 2 (ILCINNCGFFGSPATMNLCSKCYKDMJLK). Four pairs of tandem duplications were found in eggplant, and 10 SmSAP genes had collinearity with SAP genes from Arabidopsis, potato, or tomato, but only four SmSAP genes were collinear with SAP genes in the three species mentioned above. Moreover, the promoters of SmSAP genes were predicted to contain many cis-acting elements that respond to abiotic stress and hormones. A qRT-PCR analysis of the four selected SmSAP genes exhibited diverse expression levels in response to various environmental stresses. These results provided a comprehensive analysis of the SmSAP genes and lay a solid foundation for improving the understanding of the functional diversification of SAP genes under various environmental stresses in eggplant.

Genome-Wide Identification of Stress-Associated Proteins (SAPs) Encoding A20/AN1 Zinc Finger in Almond (Prunus dulcis) and Their Differential Expression during Fruit Development.

Stress-associated proteins (SAPs) are zinc finger proteins involved in the regulation of various stresses in a variety of plant species. A total of nine PdSAP genes were identified in Prunus dulcis. Phylogenetic and synteny analyses were performed to analyze the homology and evolutionary relationship of PdSAP genes. The functions of PdSAP genes were assessed by further analyses, including cis-regulatory elements, gene duplication, gene ontology, gene structure, subcellular localization, and motif pattern. This study found that PdSAP genes were unevenly distributed on chromosomes 2, 3, 6, and 7. Phylogenetic analysis of PdSAP genes with Arabidopsis thaliana and Oryza sativa suggested that six subgroups have a similar pattern of AN1 and A20 domains in each subgroup. PdSAP genes lacked duplicated blocks. The majority of PdSAP genes were localized in the nucleus region. Three hormonal and five stress cis-regulatory elements were found in the upstream promoter region of the PdSAP gene family. RNA-seq analysis revealed differential gene expression of PdSAP genes at days 12, 17, 22, 27, 32, and 37 of fruitlet development after flowering. This study identifies the SAP genes in P. dulcis and also provides insights into the expression of PdSAP genes in abnormal fruitlets with diapause atrophic growth at various developmental stages.

Comprehensive analysis of the stress associated protein (SAP) gene family in Tamarix hispida and the function of ThSAP6 in salt tolerance

Stress associated proteins (SAPs), a class of A20/AN1 zinc finger domain-containing proteins, are involved in a variety of biotic and abiotic stress responses in plants. However, little is known about the SAP gene family and their functions in Tamarix hispida. In this study, we isolated and characterized 11 SAPs from T. hispida. The expression patterns of ThSAPs were analyzed under various stresses (salt and drought) and phytohormone treatment (SA, ABA and MeJA) using real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR). Most ThSAPs exhibited transcriptional responses to abiotic stresses and phytohormones. Among these ThSAPs, ThSAP6 was significantly induced by salt stress. Gain-and loss-of-function analyses revealed that ThSAP6 was a positive regulator of salt stress response. Overexpression of ThSAP6 in T. hispida increased antioxidant enzymes activity and proline content and decreased reactive oxygen species (ROS) accumulation and cell membrane damage under salt stress, while the opposite physiological changes were observed in ThSAP6-RNAi (RNA interference) lines. This study provides a comprehensive description of the SAP gene family in T. hispida, and demonstrates that ThSAP6 is a potential candidate for biotechnological approaches to improve salt tolerance in plants.

Expression of an A20/AN1 Stress-Associated Protein from Aeluropus littoralis in Rice Deregulates Stress-Related Genes

The AlSAP gene, which has been isolated from the C4 halophyte grass Aeluropus littoralis, a member of the stress-associated protein gene family. Like other members of this family, AlSAP contains A20 and AN1 zinc-finger domains, and its transcription is induced by abiotic stresses such as salinity, drought, and temperature as well as by several hormones, including abscisic and salicylic acids. Additionally, AlSAP overexpression in rice has been demonstrated to enhance cold and drought stress tolerance. In the present study, we investigated the transcriptional deregulation of stress-related genes following ectopic expression of the AlSAP gene in two transgenic rice lines, namely, RN4 and RN5. Compared with control plants, a total of 1734 and 3650 annotated genes were found to be differentially expressed in the leaves and roots of the RN4 and RN5 lines, respectively. AlSAP transcript accumulation primed the expression of abiotic-stress-related genes involved in transcription, signaling, protein degradation, and hormonal homeostasis in rice plants grown under unstressed conditions. We hypothesized that this priming underlies the enhanced tolerance to abiotic stresses previously reported in AlSAP rice lines. Notably, AlSAP also induces transcript accumulation of genes known to be major negative regulators of pathogen response in rice. Consistently, our results showed that abiotic stress-tolerant AlSAP rice lines also exhibit enhanced susceptibility to rice blast fungus Magnaporthe oryzae.

Identification, gene expression and genetic polymorphism of zinc finger A20/AN1 stress-associated genes, HvSAP, in salt stressed barley from Kazakhstan

BackgroundA family of genes designated as the Zinc finger A20/AN1 Transcription factors encoding stress-associated proteins (SAP) are well described in Arabidopsis and rice, and include 14 AtSAP and 18 OsSAP genes that are associated with variable tolerances to multiple abiotic stresses. The SAP gene family displays a great diversity in its structure and across different plant species. The aim of this study was to identify all HvSAP genes in barley (Hordeum vulgare L.), to analyse the expression of selected genes in response to salinity in barley leaves and develop SNP marker for HvSAP12 to evaluate the association between genotypes of barley plants and their grain yield in field trials.ResultsIn our study, 17 HvSAP genes were identified in barley, which were strongly homologous to rice genes. Five genes, HvSAP5, HvSAP6, HvSAP11, HvSAP12 and HvSAP15, were found to be highly expressed in leaves of barley plants in response to salt stress in hydroponics compared to controls, using both semi-quantitative RT-PCR and qPCR analyses. The Amplifluor-like SNP marker KATU-B30 was developed and used for HvSAP12 genotyping. A strong association (R2 = 0.85) was found between KATU-B30 and grain yield production per plant of 50 F3 breeding lines originating from the cross Granal × Baisheshek in field trials with drought and low to moderate salinity in Northern and Central Kazakhstan.ConclusionsA group of HvSAP genes, and HvSAP12 in particular, play an important role in the tolerance of barley plants to salinity and drought, and is associated with higher grain yield in field trials. Marker-assisted selection with SNP marker KATU-B30 can be applied in barley breeding to improve grain yield production under conditions of abiotic stress.

Genomic characterization and expression profiles of stress-associated proteins (SAPs) in castor bean (Ricinus communis)

Stress-associated proteins (SAPs) are known as response factors to multiple abiotic and biotic stresses in plants. However, the potential physiological and molecular functions of SAPs remain largely unclear. Castor bean (Ricinus communis L.) is one of the most economically valuable non-edible woody oilseed crops, able to be widely cultivated in marginal lands worldwide because of its broad adaptive capacity to soil and climate conditions. Whether SAPs in castor bean plays a key role in adapting diverse soil conditions and stresses remains unknown. In this study, we used the castor bean genome to identify and characterize nine castor bean SAP genes (RcSAP). Structural analysis showed that castor bean SAP gene structures and functional domain types vary greatly, differing in intron number, protein sequence, and functional domain type. Notably, the AN1-C2H2–C2H2 zinc finger domain within RcSAP9 has not been often observed in other plant families. High throughput RNA-seq data showed that castor bean SAP gene profiles varied among different tissues. In addition, castor bean SAP gene expression varied in response to different stresses, including salt, drought, heat, cold and ABA and MeJA, suggesting that the transcriptional regulation of castor bean SAP genes might operate independently of each other, and at least partially independent from ABA and MeJA signal pathways. Cis-element analyses for each castor bean SAP gene showed that no common cis-elements are shared across the nine castor bean SAP genes. Castor bean SAPs were localized to different regions of cells, including the cytoplasm, nucleus, and cytomembrane. This study provides a comprehensive profile of castor bean SAP genes that advances our understanding of their potential physiological and molecular functions in regulating growth and development and their responses to different abiotic stresses.

Global transcriptome analysis of novel stress associated protein (SAP) genes expression dynamism of combined abiotic stresses in Oryza sativa (L.)

Genes encoding proteins with A20/AN1 zinc-finger domains, belonging to the stress associated protein (SAP) gene family, are present in all eukaryotes and play a decisive role in plant response to diverse physiological and molecular activities particularly on biotic and abiotic stresses (AbS). In this first and foremost study, global transcriptome analysis of members of the SAP gene family was carried out in C3 model-Oryza sativa (OsSAP) aiming at the identification of OsSAP genes activated in response to unique or Combined AbS (CAbS). Based on the available spatio-temporal and phytohormonal RNA-Seq expression profile datasets, nine OsSAP genes were filtered out and identified by a differential expression signature noted in various tissues as well as plant hormones. Comparative genome ideogram of OsSAP genes confirmed the orthologous collinearity with C4 panicoid genomes. Interactome of these genes, revealed the molecular cross-talks of OsSAP. Thus, the computational expression signature of OsSAP genes led to a better understanding of gene dynamism in diverse developmental tissues/organs. Transcriptional regulation analysis of key OsSAP genes in response to stress (drought and salinity) suggested the novel role of OsSAP1, OsSAP2, OsSAP5, OsSAP7, OsSAP8 and OsSAP11 in AbS. Altogether, the study provides deeper insights on molecular characteristics of OsSAP genes, which could be deployed further to decipher their precise functional roles in AbS responses. Communicated by Ramaswamy H. Sarma

Identification and Expression Analysis of Stress-Associated Proteins (SAPs) Containing A20/AN1 Zinc Finger in Cucumber.

Stress-associated proteins (SAPs) are a class of zinc finger proteins that confer tolerance to a variety of abiotic and biotic stresses in diverse plant species. However, in cucumber (Cucumis sativus L.), very little is known about the roles of SAP gene family members in regulating plant growth, development, and stress responses. In this study, a total of 12 SAP genes (named as CsSAP1-CsSAP12) were identified in the cucumber genome, which were unevenly distributed on six chromosomes. Gene duplication analysis detected one tandem duplication and two segmental duplication events. Phylogenetic analysis of SAP proteins from cucumber and other plants suggested that they could be divided into seven groups (sub-families), and proteins in the same group generally had the same arrangement of AN1 (ZnF-AN1) and A20 (ZnF-A20) domains. Most of the CsSAP genes were intronless and harbored a number of stress- and hormone-responsive cis-elements in their promoter regions. Tissue expression analysis showed that the CsSAP genes had a broad spectrum of expression in different tissues, and some of them displayed remarkable alteration in expression during fruit development. RT-qPCR results indicated that all the selected CsSAP genes displayed transcriptional responses to cold, drought, and salt stresses. These results enable the first comprehensive description of the SAP gene family in cucumber and lay a solid foundation for future research on the biological functions of CsSAP genes.

Genomic Analysis of Stress Associated Proteins in Soybean and the Role of GmSAP16 in Abiotic Stress Responses in Arabidopsis and Soybean.

Stress associated proteins (SAPs) containing A20/AN1 zinc finger domains have emerged as novel regulators of stress responses. In this study, 27 SAP genes were identified in soybean. The phylogenetic relationships, exon–intron structure, domain structure, chromosomal localization, putative cis-acting elements, and expression patterns of SAPs in various tissues under abiotic stresses were analyzed. Among the soybean SAP genes, GmSAP16 was significantly induced by water deficit stress, salt, and abscisic acid (ABA) and selected for further analysis. GmSAP16 was located in the nucleus and cytoplasm. The overexpression of GmSAP16 in Arabidopsis improved drought and salt tolerance at different developmental stages and increased ABA sensitivity, as indicated by delayed seed germination and stomatal closure. The GmSAP16 transgenic Arabidopsis plants had a higher proline content and a lower water loss rate and malondialdehyde (MDA) content than wild type (WT) plants in response to stresses. The overexpression of GmSAP16 in soybean hairy roots enhanced drought and salt tolerance of soybean seedlings, with higher proline and chlorophyll contents and a lower MDA content than WT. RNA inference (RNAi) of GmSAP16 increased stress sensitivity. Stress-related genes, including GmDREB1B;1, GmNCED3, GmRD22, GmDREB2, GmNHX1, and GmSOS1, showed significant expression alterations in GmSAP16-overexpressing and RNAi plants under stress treatments. These results indicate that soybean SAP genes play important roles in abiotic stress responses.

A Stress-Associated Protein, PtSAP13, From Populus trichocarpa Provides Tolerance to Salt Stress.

The growth and production of poplars are usually affected by unfavorable environmental conditions such as soil salinization. Thus, enhancing salt tolerance of poplars will promote their better adaptation to environmental stresses and improve their biomass production. Stress-associated proteins (SAPs) are a novel class of A20/AN1 zinc finger proteins that have been shown to confer plants’ tolerance to multiple abiotic stresses. However, the precise functions of SAP genes in poplars are still largely unknown. Here, the expression profiles of Populus trichocarpa SAPs in response to salt stress revealed that PtSAP13 with two AN1 domains was up-regulated dramatically during salt treatment. The β-glucuronidase (GUS) staining showed that PtSAP13 was accumulated dominantly in leaf and root, and the GUS signal was increased under salt condition. The Arabidopsis transgenic plants overexpressing PtSAP13 exhibited higher seed germination and better growth than wild-type (WT) plants under salt stress, demonstrating that overexpression of PtSAP13 increased salt tolerance. Higher activities of antioxidant enzymes were found in PtSAP13-overexpressing plants than in WT plants under salt stress. Transcriptome analysis revealed that some stress-related genes, including Glutathione peroxidase 8, NADP-malic enzyme 2, Response to ABA and Salt 1, WRKYs, Glutathione S-Transferase, and MYBs, were induced by salt in transgenic plants. Moreover, the pathways of flavonoid biosynthesis and metabolic processes, regulation of response to stress, response to ethylene, dioxygenase activity, glucosyltransferase activity, monooxygenase activity, and oxidoreductase activity were specially enriched in transgenic plants under salt condition. Taken together, our results demonstrate that PtSAP13 enhances salt tolerance through up-regulating the expression of stress-related genes and mediating multiple biological pathways.

Genome-wide identification of stress-associated proteins (SAP) with A20/AN1 zinc finger domains associated with abiotic stresses responses in Brassica napus

Stress-associated proteins (SAP) that contain the A20/AN1 zinc-finger domain play important roles in plants abiotic stress responses. However, little is known about the SAP gene family in Brassica napus (genome AnAnCnCn), was formed by recent allopolyploidy between ancestors of Brassica rapa (genome ArAr) and Brassica oleracea (genome CoCo). To explore the distribution and expression patterns of these genes, a genome-wide identification and a systematic analysis of genes in the SAP family in B. napus were performed. A total of 57 genes encoding SAP were identified, and subsequently classified into 5 groups. In contrast to results seen in other plant species, further analyses of gene structures and conserved protein motifs revealed that a large percentage of the BnaSAP (16/57) genes were A20-type or AN1-type SAP genes, most of which were not found to have homologous genes in B. rapa or B. oleracea. The cis-elements of the BnaSAP promoters, and BnaSAP gene expression levels in different tissues, and under different stress treatments, were also investigated based on RNA-seq data. All the typical A20-AN1-type and AN1-AN1(-C2H2)-type BnaSAP genes were constitutively expressed among all tissues, except 4 members in Group II. In contrast, almost all A20 type and AN1 type members showed very low detection levels in all the tested rapeseed tissues from which the transcriptome data was derived. Additionally, most of the typical A20-AN1-type and AN1-AN1-type BnaSAP genes were found to be induced by multiple stresses and phytohormones. Among the identified BnaSAP genes, four members of Group III (BnaA07g15770D, BnaC06g14100D, BnaC04g27800D, and BnaC08g24010D) were induced following various low-temperature stresses in leaves, and were induced by NaCl and PEG stress in roots. BnaA06g02460D and BnaC06g04440D from Group IV were induced by heat stress, while BnaA06g31640D and BnaC07g25010D from Group V were induced by heat stress and salicylic acid (SA) treatment. This study provides a comprehensive analysis of SAP genes in rapeseed, and may aid in future efforts to identify the functions of A20/AN1-type proteins and the responses of rapeseed under a variety of abiotic stresses.

IDENTIFICATION OF GENES ENCODING STRESS-ASSOCIATED PROTEINS CONTAINING THE DOMAINS A20/AN1 IN THE APPLE GENOME IN SILICO AND ANALYSIS OF THEIR PHYLOGENETIC RELATIONSHIP

Stress-associated proteins (SAPs) are the transcription factors containing the zinc-finger domains A20 and/or AN1. Genome-wide identification and characterization of an apple variety of Golden Delicious stress-associated proteins was performed, potential SAPs were found. Each of them contains, at least, one zinc-finger domain AN1. A phylogenetic tree was constructed on the basis of the amino acid sequences of apple SAP and homologues from other species (dicotyledonous cotton and monocotyledonous rice) to assess their phylogenetic relationships. The sequences ahead of the translation initiation site of SAPs genes were analyzed to discover cis-elements involved in the gene expression regulation. The analysis shows that apple SAPs are involved in a complex network of regulatory elements that controls the vitality of plant cells. It can be assumed that the expression of SAPs in apple trees occurs constantly, but if conditions of a plant growth are changed, their level increases. Regulation of the SAP expression may depend on a plant ontogenesis stage and it may vary in different tissues.