Halotolerant PGPB Staphylococcus sciuri ET101 protects photosynthesis through activation of redox dissipation pathways in Lycopersicon esculentum.

Photosynthesis is known to be seriously affected by salt stress. The stress induced membrane damage leads to disrupted photosynthetic components causing imbalance between production and utilization of ATP/NADPH with generation of ROS leading to photoinhibition and photodamage. In the current study, role of halotolerant plant growth promoting bacteria (PGPB) Staphylococcus sciuri ET101 in protection of photosynthesis in tomato plants during salinity stress was evaluated by analysing changes in antioxidant defense and activation of redox dissipation pathways. Inoculation of S. sciuri ET101 significantly enhanced the growth of tomato plants with significantly higher photosynthetic rates (PN) under normal and salinity stress conditions. Further, increased membrane stability, soluble sugar accumulation and significant decrease in malondialdehyde (MDA) content in leaves of ET101 inoculated tomato plants under normal and salinity were observed along with increased expression of antioxidant genes for efficient ROS detoxification and suppression of oxidative damage. Additionally, salinity induced decrease in rate of photosynthesis (PN) due to lowered chloroplastic CO2 concentration (Cc) attributed by low mesophyll conductance (gm) in uninoculated plants was alleviated by ET101 inoculation showing significantly higher carboxylation rate (Vcmax), RuBP generation (Jmax) and increased photorespiration (PR). The genes involved in photorespiratory process, cyclic electron flow (CEF), and alternative oxidase (AOX) pathway of mitochondrial respiration were abundantly expressed in leaves of ET101 inoculated plants indicating their involvement in protecting photosynthesis from salt stress induced photoinhibition. Collectively, our results indicated that S. sciuri ET101 has the potential in protecting photosynthesis of tomato plants under salinity stress through activation of redox dissipation pathways.

Exacerbation of drought-induced physiological and biochemical changes in leaves of Pisum sativum upon restriction of COX and AOX pathways of mitochondrial oxidative electron transport.

Drought stress affects photosynthesis, leading to significant decrease in crop productivity. In the current study, the importance of the cytochrome oxidase (COX) and alternative oxidase (AOX) pathways of themitochondrial oxidative electron transport chain (mETC) for photosynthesis and reactive oxygen species (ROS) homeostasis was evaluated in the leaves of Pisum sativum plants exposed to drought stress for 3 days (D3), 6 days (D6), and 9 days (D9). While drought stress resulted in decreased CO2 assimilation rates, leaf stomatal conductance, transpiration, and leaf intercellular CO2 concentration in a stress-dependent manner, superimposition with mETC inhibitors, antimycin A (AA) and salicylhydroxamic acid (SHAM), aggravated the responses. Decreased chlorophyll content, photosynthesis, and RubisCO (RbcL) degradation during progressive drought and their aggravation upon AOX pathway restriction indicated the importance of the AOX pathway for photosynthetic activity. Compared with COX pathway inhibition, higher intracellular H2O2 and O2.- levels, and increased cell death upon restriction of the AOX pathway during D6 and D9 stress conditions correlating with the modulation in antioxidant enzyme activities, signify the essentiality of the AOX pathway for ROS maintenance at optimal levels. Further, increased AOX1a expression during D6 and D9 conditions along with increasedAOXprotein levels indicated the activation of theAOXpathway during drought stress. Decline in Fv/Fm, actual quantum yield of PSII (ФPSII), photochemical quenching (qP), non-photochemical quenching (NPQ), and electron transport rate (ETR) upon restriction of the COX and AOX pathways indicated the requirement of mETC activity for optimal photochemical activities not only under normal conditions but also under progressive drought conditions.

Trehalose accumulation enhances drought tolerance by modulating photosynthesis and ROS-antioxidant balance in drought sensitive and tolerant rice cultivars.

Trehalose being an integral part for plant growth, development and abiotic stress tolerance is accumulated in minute amounts in angiosperms with few exceptions from resurrection plants. In the current study, two rice cultivars differing in drought tolerance were used to analyse the role of trehalose in modulating photosynthesis and ROS-antioxidant balance leading to improvement in drought tolerance. Accumulation of trehalose in leaves of Vaisakh (drought-tolerant) and Aiswarya (drought-sensitive) rice cultivars was observed by spraying 50 mM trehalose and 100 µM validamycin A (trehalase inhibitor) followed by vacuum infiltration. Compared to stress sensitive Aiswarya cultivar, higher trehalose levels were observed in leaves of Vaisakh not only under control conditions but also under drought conditions corresponding with increased root length. The increase in leaf trehalose by treatment with trehalose or validamycin A corresponded well with a decrease in electrolyte leakage in sensitive and tolerant plants. Decreased ROS levels were reflected as increase in antioxidant enzyme activity and their gene expression in leaves of both the cultivars treated with trehalose or Validamycin A under control and drought conditions signifying the importance of trehalose in modulating the ROS-antioxidant balance for cellular protection. Further, higher chlorophyll, higher photosynthetic activity and modulation in other gas exchange parameters upon treatment with trehalose or validamycin A strongly suggested the beneficial role of trehalose for stress tolerance. Trehalose accumulation helped the tolerant cultivar adjust towards drought by maintaining higher water status and alleviating the ROS toxicity by effective activation and increment in antioxidant enzyme activity along with enhanced photosynthesis. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-023-01404-7.

Biofilm-Associated Agr and Sar Quorum Sensing Systems of Staphylococcus aureus Are Inhibited by 3-Hydroxybenzoic Acid Derived from Illicium verum.

Biofilm-producing Staphylococcus aureus (S. aureus) is less sensitive to conventional antibiotics than free-living planktonic cells. Here, we evaluated the antibiofilm activity of Illicium verum (I. verum) and one of its constituent compounds 3-hydroxybenzoic acid (3-HBA) against multi-drug-resistant S. aureus. We performed gas chromatography-mass spectroscopy (GC-MS) to identify the major constituents in the methanolic extract of I. verum. Ligand-receptor interactions were studied by molecular docking, and in vitro investigations were performed using crystal violet assay, spreading assay, hemolysis, proteolytic activity, and growth curve analysis. The methanolic extract of I. verum inhibited S. aureus at 4.8 mg/mL, and GC-MS analysis revealed anethole, m-methoxybenzaldehyde, and 3-HBA as the major constituents. Molecular docking attributed the antibiofilm activity to an active ligand present in 3-HBA, which strongly interacted with the active site residues of AgrA and SarA of S. aureus. At a subinhibitory concentration of 2.4 mg/mL, the extract showed biofilm inhibition. Similarly, 3-HBA inhibited biofilm activity at 25 µg/mL (90.34%), 12.5 µg/mL (77.21%), and 6.25 µg/mL (62.69%) concentrations. Marked attrition in bacterial spreading was observed at 2.4 mg/mL (crude extract) and 25 µg/mL (3-HBA) concentrations. The methanol extract of I. verum and 3-HBA markedly inhibited ß-hemolytic and proteolytic activities of S. aureus. At the lowest concentration, the I. verum extract (2.4 mg/mL) and 3-HBA (25 µg/mL) did not inhibit bacterial growth. Optical microscopy and SEM analysis confirmed that I. verum and 3-HBA significantly reduced biofilm dispersion without disturbing bacterial growth. Together, we found that the antibiofilm activity of I. verum and 3-HBA strongly targeted the Agr and Sar systems of S. aureus.

Differential modulation of photosynthesis, ROS and antioxidant enzyme activities in stress-sensitive and -tolerant rice cultivars during salinity and drought upon restriction of COX and AOX pathways of mitochondrial oxidative electron transport.

Drought and salt stresses are two major abiotic stress factors that hamper crop growth and productivity. Three rice cultivars with different sensitivity and tolerance towards abiotic stress were used in the current study. While cultivar Aiswarya is salt- and drought-sensitive, cultivar Vyttila is salt-tolerant and cultivar Vaisakh is drought-tolerant. We compared the physiological and biochemical responses of these rice cultivars under salt and drought stress conditions after restricting their cytochrome oxidase (COX) and alternative oxidase (AOX) pathways using antimycin A and salicylhydroxamic acid treatment. Further, changes in their expression of AOX genes and corresponding protein levels were compared and analysed. The sensitive and tolerant rice cultivars subjected to drought and salt stress showed differential responses in physiological and biochemical traits. Whereas Aiswarya showed clear phenotypic differences, such as stunted growth, leaf curling, and loss of greening in leaf tissues, with increase in salt content and progressive drought stress, Vyttila and Vaisakh showed no remarkable changes. Moreover, the drought-tolerant cultivar rehydrated after 10 days of drought exposure, whereas the sensitive variety did not show any rehydration of leaf tissue. The leaves of the tolerant cultivars showed lower reactive oxygen species (ROS) production than that of the sensitive plants under drought and salt stress conditions because of the activation of a stronger antioxidant defence. Although, the restriction of COX and AOX pathways increased the susceptibility of sensitive cultivars, it affected the tolerant varieties moderately. Higher photosynthetic rates, an efficient antioxidant system comprising higher superoxide dismutase, ascorbate peroxidase, and catalase activity along with higher AOX1a gene expression levels during drought and salt stress were observed in tolerant cultivars. The results suggest that an efficient antioxidant system and increased transcription of the AOX1a gene along with higher AOX protein levels are important for tolerant rice cultivars to maintain higher photosynthesis rates, lower ROS, and stress tolerance. Restriction of COX and AOX pathways impact the photosynthesis, ROS, and antioxidant enzymes in both sensitive and tolerant cultivars. The restriction of COX and AOX pathways have a stronger impact on gas exchange and fluorescence parameters of the sensitive cultivar than on that of the tolerant cultivars owing to the higher photosynthetic rates in tolerant cultivars.

Molecular insights into plant desiccation tolerance: transcriptomics, proteomics and targeted metabolite profiling in Craterostigma plantagineum.

The resurrection plant Craterostigma plantagineum possesses an extraordinary capacity to survive long-term desiccation. To enhance our understanding of this phenomenon, complementary transcriptome, soluble proteome and targeted metabolite profiling was carried out on leaves collected from different stages during a dehydration and rehydration cycle. A total of 7348 contigs, 611 proteins and 39 metabolites were differentially abundant across the different sampling points. Dynamic changes in transcript, protein and metabolite levels revealed a unique signature characterizing each stage. An overall low correlation between transcript and protein abundance suggests a prominent role for post-transcriptional modification in metabolic reprogramming to prepare plants for desiccation and recovery. The integrative analysis of all three data sets was performed with an emphasis on photosynthesis, photorespiration, energy metabolism and amino acid metabolism. The results revealed a set of precise changes that modulate primary metabolism to confer plasticity to metabolic pathways, thus optimizing plant performance under stress. The maintenance of cyclic electron flow and photorespiration, and the switch from C3 to crassulacean acid metabolism photosynthesis, may contribute to partially sustain photosynthesis and minimize oxidative damage during dehydration. Transcripts with a delayed translation, ATP-independent bypasses, alternative respiratory pathway and 4-aminobutyric acid shunt may all play a role in energy management, together conferring bioenergetic advantages to meet energy demands upon rehydration. This study provides a high-resolution map of the changes occurring in primary metabolism during dehydration and rehydration and enriches our understanding of the molecular mechanisms underpinning plant desiccation tolerance. The data sets provided here will ultimately inspire biotechnological strategies for drought tolerance improvement in crops.

Cytochrome oxidase and alternative oxidase pathways of mitochondrial electron transport chain are important for the photosynthetic performance of pea plants under salinity stress conditions.

The flexible plant mitochondrial electron transport chain with cytochrome c oxidase (COX) and alternative oxidase (AOX) pathways is known to be modulated by abiotic stress conditions. The effect of salinity stress on the mitochondrial electron transport chain and the importance of COX and AOX pathways for optimization of photosynthesis under salinity stress conditions is not clearly understood. In the current study, importance of COX and AOX pathways for photosynthetic performance of pea plants (Pisum sativum L. Pea Arkel cv) was analysed by using the mitochondrial electron transport chain inhibitors Antimycin A (AA) and salicylhydroxamic acid (SHAM) which restrict the electron flow through COX and AOX pathways respectively. Salinity stress resulted in decreased CO2 assimilation rates, leaf stomatal conductance, transpiration and leaf intercellular CO2 concentration in a stress dependent manner. Superimposition of leaves of salt stressed plants with AA and SHAM caused cellular H2O2 and O2- accumulation along with cell death. Additionally, aggravation in decrease of CO2 assimilation rates, leaf stomatal conductance, transpiration and leaf intercellular CO2 concentration upon superimposition with AA and SHAM during salinity stress suggests the importance of mitochondrial oxidative electron transport for photosynthesis. Increased expression of AOX1a and AOX2 transcripts along with AOX protein levels indicated up regulation of AOX pathway in leaves during salinity stress. Chlorophyll fluorescence measurements revealed enhanced damage to Photosystem (PS) II in the presence of AA and SHAM during salinity stress. Results suggested the beneficial role of COX and AOX pathways for optimal photosynthetic performance in pea leaves during salinity stress conditions.

Protection of Photosynthesis by Halotolerant Staphylococcus sciuri ET101 in Tomato (Lycoperiscon esculentum) and Rice (Oryza sativa) Plants During Salinity Stress: Possible Interplay Between Carboxylation and Oxygenation in Stress Mitigation.

Tomato (Lycoperiscon esculentum) and rice (Oryza sativa) are the two most important agricultural crops whose productivity is severely impacted by salinity stress. Soil salinity causes an irreversible damage to the photosynthetic apparatus in plants at all developmental stages leading to significant reduction in agricultural productivity. Reduction in photosynthesis is the primary response that is observed in all glycophytic plants during salt stress. Employment of salt-tolerant plant growth-promoting bacteria (PGPB) is an economical and viable approach for the remediation of saline soils and improvement of plant growth. The current study is aimed towards investigating the growth patterns and photosynthetic responses of rice and tomato plants upon inoculation with halotolerant PGPB Staphylococcus sciuri ET101 under salt stress conditions. Tomato and rice plants inoculated with PGPB showed increased growth rate and stimulated root growth, along with higher transpiration rates (E), stomatal conductance (g s ), and intracellular CO2 accumulation (Ci). Additionally, correlation of relative water content (RWC) to electrolyte leakage (EL) in tomato and rice plants showed decreased EL in inoculated plants during salt stress conditions, along with higher proline and glycine betaine content. Energy dissipation by non-photochemical quenching (NPQ) and increased photorespiration of 179.47% in tomato and 264.14% in rice plants were observed in uninoculated plants subjected to salinity stress. Furthermore, reduced photorespiration with improved salinity tolerance is observed in inoculated plants. The higher rates of photosynthesis in inoculated plants during salt stress were accompanied by increased quantum efficiency (ΦPSII) and maximum quantum yield (F v /F m ) of photosystem II. Furthermore, inoculated plants showed increased carboxylation efficiency of RuBisCO, along with higher photosynthetic electron transport rate (ETR) (J) during salinity stress. Although the total cellular ATP levels are drastically affected by salt stress in tomato and rice plants along with increased reactive oxygen species (ROS) accumulation, the restoration of cellular ATP levels in leaves of inoculated plants along with decreased ROS accumulation suggests the protective role of PGPB. Our results reveal the beneficial role of S. sciuri ET101 in protection of photosynthesis and amelioration of salinity stress responses in rice and tomato plants.

UV-B priming of Oryza sativa var. Kanchana seedlings augments its antioxidative potential and gene expression of stress-response proteins under various abiotic stresses.

Priming is one of the mechanisms for the induction of the antioxidant defense system and various stress-responsive proteins which help plants to survive under various abiotic stresses. Based on the observation that the rice seedlings primed with UV-B (low dose of UV-B irradiation-6 kJm-2) induced the acclimation against NaCl, PEG and UV-B stresses, it was of interest to see the augmentation of antioxidative potential and stress-responsive proteins accumulation in rice seedlings due to UV-B priming under these stresses. Various stresses result in production of ROS, which cause membrane degradation resulting in the accumulation of malondialdehyde. These negative impacts were observed exceedingly in rice seedlings from non-primed PEG stress (NP+P) condition than UV-B and NaCl stresses. The production of non-enzymatic antioxidants, activity/mRNA-level expressions of enzymatic antioxidants and stress-responsive proteins were effectively augmented in UV-B-primed rice seedlings subjected to NaCl stress (P+N) condition followed by UV-B stress (P+U) and PEG stress (P+P). The activation of stress-responsive proteins (HSP and LEA) in rice due to the UV-B priming of rice seedlings is being reported for the first time. The results revealed that the UV-B seedling priming was alleviating the effect of NaCl, PEG, and UV-B stresses in rice seedlings. The positive impacts of UV-B seedling priming were more prominent in rice seedlings subjected to NaCl stress, indicating the cross tolerance imparted by UV-B priming.

Transcriptional and metabolic changes in the desiccation tolerant plant Craterostigma plantagineum during recurrent exposures to dehydration.

MAIN CONCLUSION: Multiple dehydration/rehydration treatments improve the adaptation of Craterostigma plantagineum to desiccation by accumulating stress-inducible transcripts, proteins and metabolites. These molecules serve as stress imprints or memory and can lead to increased stress tolerance. It has been reported that repeated exposure to dehydration may generate stronger reactions during a subsequent dehydration treatment in plants. This stimulated us to address the question whether the desiccation tolerant resurrection plant Craterostigma plantagineum has a stress memory. The expression of four representative stress-related genes gradually increased during four repeated dehydration/rehydration treatments in C. plantagineum. These genes reflect a transcriptional memory and are trainable genes. In contrast, abundance of chlorophyll synthesis/degradation-related transcripts did not change during dehydration and remained at a similar level as in the untreated tissues during the recovery phase. During the four dehydration/rehydration treatments the level of ROS pathway-related transcripts, superoxide dismutase (SOD) activity, proline, and sucrose increased, whereas H2O2 content and electrolyte leakage decreased. Malondialdehyde (MDA) content did not change during the dehydration, which indicates a gain of stress tolerance. At the protein level, increased expression of four representative stress-related proteins showed that the activated stress memory can persist over several days. The phenomenon described here could be a general feature of dehydration stress memory responses in resurrection plants.

Protection of photosynthesis in desiccation-tolerant resurrection plants.

Inhibition of photosynthesis is a central, primary response that is observed in both desiccation-tolerant and desiccation-sensitive plants affected by drought stress. Decreased photosynthesis during drought stress can either be due to the limitation of carbon dioxide entry through the stomata and the mesophyll cells, due to increased oxidative stress or due to decreased activity of photosynthetic enzymes. Although the photosynthetic rates decrease in both desiccation-tolerant and sensitive plants during drought, the remarkable difference lies in the complete recovery of photosynthesis after rehydration in desiccation-tolerant plants. Desiccation of sensitive plants leads to irreparable damages of the photosynthetic membranes, in contrast the photosynthetic apparatus is deactivated during desiccation in desiccation-tolerant plants. Desiccation-tolerant plants employ different strategies to protect and/or maintain the structural integrity of the photosynthetic apparatus to reactivate photosynthesis upon water availability. Two major mechanisms are distinguished. Homoiochlorophyllous desiccation-tolerant plants preserve chlorophyll and thylakoid membranes and require active protection mechanisms, while poikilochlorophyllous plants degrade chlorophyll in a regulated manner but then require de novo synthesis during rehydration. Desiccation-tolerant plants, particularly homoiochlorophyllous plants, employ conserved and novel antioxidant enzymes/metabolites to minimize the oxidative damage and to protect the photosynthetic machinery. De novo synthesized, stress-induced proteins in combination with antioxidants are localized in chloroplasts and are important components of the protective network. Genome sequence informations provide some clues on selection of genes involved in protecting photosynthetic structures; e.g. ELIP genes (early light inducible proteins) are enriched in the genomes and more abundantly expressed in homoiochlorophyllous desiccation-tolerant plants. This review focuses on the mechanisms that operate in the desiccation-tolerant plants to protect the photosynthetic apparatus during desiccation.

Surviving metabolic arrest: photosynthesis during desiccation and rehydration in resurrection plants.

Photosynthesis is the key process that is affected by dehydration in plants. Desiccation-tolerant resurrection plants can survive conditions of very low relative water content. During desiccation, photosynthesis is not operational, but is recovered within a short period after rehydration. While homoiochlorophyllous resurrection plants retain their photosynthetic apparatus during desiccation, poikilochlorophyllous resurrection species dismantle chloroplasts and degrade chlorophyll but resynthesize them again during rehydration. Dismantling the chloroplasts avoids the photooxidative stress in poikilochlorophyllous resurrection plants, whereas it is minimized in homoiochlorophyllous plants through the synthesis of antioxidant enzymes and protective proteins or metabolites. Although the cellular protection mechanisms in both of these species vary, these mechanisms protect cells from desiccation-induced damage and restore photosynthesis upon rehydration. Several of the proteins synthesized during dehydration are localized in chloroplasts and are believed to play major roles in the protection of photosynthetic structures and in recovery in resurrection species. This review focuses on the strategies of resurrection plants in terms of how they protect their photosynthetic apparatus from oxidative stress during desiccation without membrane damage and with full recovery during rehydration. We review the role of the dehydration-induced protection mechanisms in chloroplasts and how photosynthesis is restored during rehydration.

Differences in LEA-like 11-24 gene expression in desiccation tolerant and sensitive species of Linderniaceae are due to variations in gene promoter sequences.

Many desiccation induced late embryogenesis abundant (LEA) protein encoding genes have been identified from Craterostigma plantagineum Hochst. In the desiccation tolerant plants C. plantagineum (Cp) and Lindernia brevidens Skan (Lb) transcripts encoding LEA-like 11-24 protein are abundantly expressed during desiccation whereas in Lindernia subracemosa De Wild. (Ls), a desiccation sensitive plant, the LEA-like 11-24 transcripts are expressed at a low level. Since promoters determine gene expression, a comparative promoter analysis was carried out to decipher the underlying mechanisms of differential gene expression. Two transient transformation methods (particle bombardment and optimised Agrobacterium co-cultivation) were used to analyse the promoter activities of the Cp, Lb and Ls LEA-like 11-24 gene in homologous and heterologous systems. Minimal promoters were isolated from all three species and their promoter activities were assessed in response to mannitol or ABA. Particle bombardment or Agrobacterium co-cultivation yielded similar results. Site-directed mutagenesis was used to identify which cis-acting elements in the LEA-like 11-24 promoter fragments are crucial during mannitol and ABA treatments. The presence of these promoter cis-elements explains the differences in transcript abundance in the desiccation tolerant and desiccation sensitive species. Results indicated the importance of the drought responsive elements (DRE) element for promoter activity.

Single-molecule sequencing of the desiccation-tolerant grass Oropetium thomaeum.

Plant genomes, and eukaryotic genomes in general, are typically repetitive, polyploid and heterozygous, which complicates genome assembly. The short read lengths of early Sanger and current next-generation sequencing platforms hinder assembly through complex repeat regions, and many draft and reference genomes are fragmented, lacking skewed GC and repetitive intergenic sequences, which are gaining importance due to projects like the Encyclopedia of DNA Elements (ENCODE). Here we report the whole-genome sequencing and assembly of the desiccation-tolerant grass Oropetium thomaeum. Using only single-molecule real-time sequencing, which generates long (>16 kilobases) reads with random errors, we assembled 99% (244 megabases) of the Oropetium genome into 625 contigs with an N50 length of 2.4 megabases. Oropetium is an example of a 'near-complete' draft genome which includes gapless coverage over gene space as well as intergenic sequences such as centromeres, telomeres, transposable elements and rRNA clusters that are typically unassembled in draft genomes. Oropetium has 28,466 protein-coding genes and 43% repeat sequences, yet with 30% more compact euchromatic regions it is the smallest known grass genome. The Oropetium genome demonstrates the utility of single-molecule real-time sequencing for assembling high-quality plant and other eukaryotic genomes, and serves as a valuable resource for the plant comparative genomics community.

Quantification of expression of dehydrin isoforms in the desiccation tolerant plant Craterostigma plantagineum using specifically designed reference genes.

Craterostigma plantagineum is a desiccation tolerant resurrection plant. Many genes are induced during desiccation. Dehydrins are a group of dehydration-induced genes present in all higher plants. The current study aims at classifying the most abundantly expressed dehydrin genes from vegetative tissues of C. plantagineum and quantifying their expression. To identify variations between dehydrin isoforms at different stages of desiccation and rehydration by RT-qPCR, the target mRNA requires an accurate and reliable normalization. Previously we reported that RNAs from leaves and roots of C. plantagineum are not degraded during desiccation and subsequent rehydration thus allowing the use of RT-qPCR to test the stability of reference genes. The expression stability of eight candidate reference genes was tested in leaves, roots and callus. These genes were ranked according to their stability of gene expression using GeNorm(PLUS) and RefFinder. The most consistently expressed reference genes in each tissue were identified and used to normalize gene expression data. Dehydrin isoforms were divided in three groups based on the expression level during the desiccation process in three different tissues (leaves, roots and callus).