ABA Signaling Research Articles

Ectopic expression of DnaJ type-I protein homolog of Vigna aconitifolia (VaDJI) confers ABA insensitivity and multiple stress tolerance in transgenic tobacco plants.

Reduced crop productivity results from altered plant physiological processes caused by dysfunctional proteins due to environmental stressors. In this study, a novel DnaJ Type-I encoding gene, VaDJI having a zinc finger motif in its C-terminal domain was found to be induced early upon treatment with heat stress (within 5min) in a heat tolerant genotype of Vigna aconitifolia RMO-40. VaDJI is induced by multiple stresses. In tobacco, ectopic expression of VaDJI reduced ABA sensitivity during seed germination and the early stages of seedling growth of transgenic tobacco plants. Concomitantly, it also improved the ability of transgenic tobacco plants to withstand drought stress by modulating the photosynthetic efficiency, with the transgenic plants having higher Fv/Fm ratios and reduced growth inhibition. Additionally, transgenic plants showed a reduced build-up of H2O2 and lower MDA levels and higher chlorophyll content during drought stress, which attenuated cell damage and reduced oxidative damage. An analysis using the qRT-PCR study demonstrated that VaDJI overexpression is associated with the expression of some ROS-detoxification-related genes and stress-marker genes that are often induced during drought stress responses. These findings suggest a hypothesis whereby VaDJI positively influences drought stress tolerance and ABA signalling in transgenic tobacco, and suggests that it is a potential gene for genetic improvement of drought and heat stress tolerance in crop plants.

GhMYB44 enhances stomatal closure to confer drought stress tolerance in cotton and Arabidopsis

MYB genes play crucial roles in plant response to abiotic stress. However, the function of MYB genes in cotton during abiotic stress is less well elucidated. Here, we found an R2R3-type MYB gene, GhMYB44, was induced by simulated drought (PEG6000) and ABA in three cotton varieties. After drought stress, the GhMYB44-silenced plants showed substantial changes at the physiological level, including significantly increased malondialdehyde content and decreased SOD activity. Silencing the GhMYB44 gene increased stomatal aperture and water loss rate, reduced plant drought tolerance. Transgenic Arabidopsis thaliana over-expressed GhMYB44 (GhMYB44-OE) enhanced resistance to mannitol-simulated osmotic stress. The stomatal aperture of the GhMYB44-OE Arabidopsis was significantly smaller than those of the wild type (WT), and the GhMYB44-OE Arabidopsis increased tolerance to drought stress. Transgenic Arabidopsis had higher germination rate under ABA treatment compared to WT, and the transcript levels of AtABI1, AtPP2CA and AtHAB1 were suppressed in GhMYB44-OE plants, indicating a potential role of GhMYB44 in the ABA signal pathway. These results showed that GhMYB44 acts as a positive regulator in plant response to drought stress, potentially useful for engineering drought-tolerant cotton.

Genome-Wide Investigation and Co-Expression Network Analysis of SBT Family Gene in Gossypium.

Subtilases (SBTs), which belong to the serine peptidases, control plant development by regulating cell wall properties and the activity of extracellular signaling molecules, and affect all stages of the life cycle, such as seed development and germination, and responses to biotic and abiotic environments. In this study, 146 Gossypium hirsutum, 138 Gossypium barbadense, 89 Gossypium arboreum and 84 Gossypium raimondii SBTs were identified and divided into six subfamilies. Cotton SBTs are unevenly distributed on chromosomes. Synteny analysis showed that the members of SBT1 and SBT4 were expanded in cotton compared to Arabidopsis thaliana. Co-expression network analysis showed that six Gossypium arboreum SBT gene family members were in a network, among which five SBT1 genes and their Gossypium hirsutum and Arabidopsis thaliana direct homologues were down-regulated by salt treatment, indicating that the co-expression network might share conserved functions. Through co-expression network and annotation analysis, these SBTs may be involved in the biological processes of auxin transport, ABA signal transduction, cell wall repair and root tissue development. In summary, this study provides valuable information for the study of SBT genes in cotton and excavates SBT genes in response to salt stress, which provides ideas for cotton breeding for salinity resistance.

Multiple-axle box acceleration measurements at railway transition zones

This paper presents a methodology for monitoring transition zones using responses of multiple-axle box acceleration (multi-ABA) measurements. The time–frequency characteristics of the vertical ABA signals from four wheelsets are analyzed. The major contributions are as follows. (1) We propose four key performance indicators (KPIs) to quantify local multi-ABA energy differences at different abutments, tracks, entrance and exit sides, and inner and outer rails. (2) The same dominant spatial frequencies are obtained with different measurement speeds, so the proposed method is suitable for multi-ABA systems mounted on operational trains. Transition zones at nine double-track railway bridges are selected as the case study. The KPIs indicate that (1) the energy differences between abutments are above 80% in three bridges; (2) two abutments show that the energy differences between tracks are higher than 100%; (3) three tracks have energy differences above 100% between the entrance and exit sides; and (4) the energy differences between rails are above 80% on three sides. Finally, using measurement with 7 years of difference, the KPIs and track quality index are discussed. These findings suggest that multi-ABA measurement can be used as a health condition monitoring method for railway transition zones to support condition-based maintenance.

Multifaceted regulatory functions of CsBPC2 in cucumber under salt stress conditions.

BASIC PENTACYSTEINE (BPC) transcription factors are essential regulators of plant growth and development. However, BPC functions and the related molecular mechanisms during cucumber (Cucumis sativus L.) responses to abiotic stresses, especially salt stress, remain unknown. We previously determined that salt stress induces CsBPC expression in cucumber. In this study, Csbpc2 transgene-free cucumber plants were created using a CRISPR/Cas9-mediated editing system to explore CsBPC functions associated with the salt stress response. The Csbpc2 mutants had a hypersensitive phenotype, with increased leaf chlorosis, decreased biomass, and increased malondialdehyde and electrolytic leakage levels under salt stress conditions. Additionally, a mutated CsBPC2 resulted in decreased proline and soluble sugar contents and antioxidant enzyme activities, which led to the accumulation of hydrogen peroxide and superoxide radicals. Furthermore, the mutation to CsBPC2 inhibited salinity-induced PM-H+-ATPase and V-H+-ATPase activities, resulting in decreased Na+ efflux and increased K+ efflux. These findings suggest that CsBPC2 may mediate plant salt stress resistance through its effects on osmoregulation, reactive oxygen species scavenging, and ion homeostasis-related regulatory pathways. However, CsBPC2 also affected ABA signaling. The mutation to CsBPC2 adversely affected salt-induced ABA biosynthesis and the expression of ABA signaling-related genes. Our results indicate that CsBPC2 may enhance the cucumber response to salt stress. It may also function as an important regulator of ABA biosynthesis and signal transduction. These findings will enrich our understanding of the biological functions of BPCs, especially their roles in abiotic stress responses, thereby providing the theoretical basis for improving crop salt tolerance.

Integrative Analysis of the Transcriptome and Metabolome Reveals the Developmental Mechanisms and Metabolite Biosynthesis of the Tuberous Roots of Tetrastigma hemsleyanum.

The tuberous root of Tetrastigma hemsleyanum Diels et Gilg (T. hemsleyanum) is a traditional Chinese medicine with a wide range of clinical applications. However, the scarcity of its wild resources, its low yield, and the variable quality that results from its artificial cultivation leads to expensive market prices that are not conducive to the further industrial development of T. hemsleyanum. In this study, transcriptomic and non-targeted metabolomic analyses were integrated to explore the underlying molecular mechanisms and metabolite biosynthesis that occur during its root development. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis revealed that differentially expressed genes (DEGs) were predominantly enriched for processes associated with flavonoid and phenylpropanoid biosynthesis, starch and sucrose metabolism, and plant hormone signal transduction. Genes related to lignin were downregulated in tuberous roots (TRs), resulting in a decrease in lignification and the downregulation of metabolites related to flavonoids and phenylpropanoid biosynthesis. In addition, the expression levels of starch- and sucrose-related genes were upregulated in TRs. The root development of SYQ is also related to IAA, GA, ABA, and JA signaling pathways. Collectively, this study lays the foundation for analyzing the root development and quality-modulating mechanisms employed by T. hemsleyanum; this will be beneficial in conducting molecular-assisted breeding and controlling its secondary metabolite production.

Integration of light and ABA signaling pathways to combat drought stress in plants.

Drought is one of the most critical stresses, which causes an enormous reduction in crop yield. Plants develop various strategies like drought escape, drought avoidance, and drought tolerance to cope with the reduced availability of water during drought. Plants adopt several morphological and biochemical modifications to fine-tune their water-use efficiency to alleviate drought stress. ABA accumulation and signaling plays a crucial role in the response of plants towards drought. Here, we discuss how drought-induced ABA regulates the modifications in stomatal dynamics, root system architecture, and the timing of senescence to counter drought stress. These physiological responses are also regulated by light, indicating the possibility of convergence of light- and drought-induced ABA signaling pathways. In this review, we provide an overview of investigations reporting light-ABA signaling crosstalk in Arabidopsis as well as other crop species. We have also tried to describe the potential role of different light components and their respective photoreceptors and downstream factors like HY5, PIFs, BBXs, and COP1 in modulating drought stress responses. Finally, we highlight the possibilities of enhancing the plant drought resilience by fine-tuning light environment or its signaling components in the future.

PhebZIP47, a bZIP transcription factor from moso bamboo (Phyllostachys edulis), positively regulates the drought tolerance of transgenic plants

The bZIP transcription factor (TF) family plays indispensable roles in plant response to environmental stresses. However, little is known about the functional roles of bZIP genes in moso bamboo (Phyllostachys edulis). Here, transcriptome data from public databases was used to analyse the expression patterns of 154 bZIP genes, and the results exhibited that the expression of many bZIP genes were strongly induced by drought. Combined with weighted gene co-expression network and qRT-PCR analysis, we screened a key candidate gene PhebZIP47 in response to drought and ABA. Next, the lines of transgenic PhebZIP47-overexpressing Arabidopsis and rice were generated. Transgenic plants showed enhanced resistance to drought at the seed germination and adult stage. Measurements of physiological indicators including chlorophyll content, relative water content, electrical conductivity, malondialdehyde and proline content were used to support this conclusion. Transcriptome analysis of transgenic rice showed that there were a large number of differentially expressed genes (DEGs) regulated by PhebZIP47, including genes of drought tolerance regulatory pathway and ABA signalling pathway. Meanwhile, a G-box element was significantly enriched in the promoters of the DEGs annotated as ‘response to stress’, and EMSA experiment suggested that this element could be bound by PhebZIP47. In addition, transgenic plants were less sensitive to ABA compared with wild-type plants under exogenous ABA treatment. Overall, our study provided a key candidate gene for the molecular breeding of moso bamboo and laid a foundation for the study of drought-resistance mechanism of bZIP genes.

The CCCH-Type Zinc-Finger Protein GhC3H20 Enhances Salt Stress Tolerance in Arabidopsis thaliana and Cotton through ABA Signal Transduction Pathway

The CCCH zinc-finger protein contains a typical C3H-type motif widely existing in plants, and it plays an important role in plant growth, development, and stress responses. In this study, a CCCH zinc-finger gene, GhC3H20, was isolated and thoroughly characterized to regulate salt stress in cotton and Arabidopsis. The expression of GhC3H20 was up-regulated under salt, drought, and ABA treatments. GUS activity was detected in the root, stem, leaves, and flowers of ProGhC3H20::GUS transgenic Arabidopsis. Compared with the control, the GUS activity of ProGhC3H20::GUS transgenic Arabidopsis seedlings under NaCl treatment was stronger. Through the genetic transformation of Arabidopsis, three transgenic lines of 35S-GhC3H20 were obtained. Under NaCl and mannitol treatments, the roots of the transgenic lines were significantly longer than those of the wild-type (WT) Arabidopsis. The leaves of the WT turned yellow and wilted under high-concentration salt treatment at the seedling stage, while the leaves of the transgenic Arabidopsis lines did not. Further investigation showed that compared with the WT, the content of catalase (CAT) in the leaves of the transgenic lines was significantly higher. Therefore, compared with the WT, overexpression of GhC3H20 enhanced the salt stress tolerance of transgenic Arabidopsis. A virus-induced gene silencing (VIGS) experiment showed that compared with the control, the leaves of pYL156-GhC3H20 plants were wilted and dehydrated. The content of chlorophyll in pYL156-GhC3H20 leaves was significantly lower than those of the control. Therefore, silencing of GhC3H20 reduced salt stress tolerance in cotton. Two interacting proteins (GhPP2CA and GhHAB1) of GhC3H20 have been identified through a yeast two-hybrid assay. The expression levels of PP2CA and HAB1 in transgenic Arabidopsis were higher than those in the WT, and pYL156-GhC3H20 had expression levels lower than those in the control. GhPP2CA and GhHAB1 are the key genes involved in the ABA signaling pathway. Taken together, our findings demonstrate that GhC3H20 may interact with GhPP2CA and GhHAB1 to participate in the ABA signaling pathway to enhance salt stress tolerance in cotton.

Exogenous Melatonin Alters Stomatal Regulation in Tomato Seedlings Subjected to Combined Heat and Drought Stress through Mechanisms Distinct from ABA Signaling

The understanding of stomatal regulation in climate stress is essential for ensuring resilient crops. The investigation of the stomatal regulation in combined heat and drought stress aimed to link effects of exogenous melatonin on stomatal conductance (gs) and its mechanistic interactions with ABA or ROS signaling. Melatonin-treated and non-treated tomato seedlings were subjected to moderate and severe levels of heat (38°C for one or three days) and drought stress (soil relative water content of 50% or 20%) applied individually and in combination. We measured gs, stomatal anatomy, ABA metabolites and enzymatic ROS scavengers. The stomata in combined stress responded predominantly to heat at soil relative water content (SRWC) = 50% and to drought stress at SRWC = 20%. Drought stress increased ABA levels at severe stress, whereas heat stress caused an accumulation of the conjugated form, ABA glucose ester, at both moderate and severe stress. The melatonin treatment affected gs and the activity of ROS scavenging enzymes but had no effect on ABA levels. The ABA metabolism and conjugation of ABA might play a role in stomatal opening toward high temperatures. We provide evidence that melatonin increases gs in combined heat and drought stress, but the effect is not mediated through ABA signaling.

Transcriptome analysis of drought-responsive and drought-tolerant mechanisms in maize leaves under drought stress.

Maize is a major crop essential for food and feed, but its production is threatened by various biotic and abiotic stresses. Drought is one of the most common abiotic stresses, causing severe crop yield reduction. Although several studies have been devoted to selecting drought-tolerant maize lines and detecting the drought-responsive mechanism of maize, the transcriptomic differences between drought-tolerant and drought-susceptible maize lines are still largely unknown. In our study, RNA-seq was performed on leaves of the drought-tolerant line W9706 and the drought-susceptible line B73 after drought treatment. We identified 3147 differentially expressed genes (DEGs) between these two lines. The upregulated DEGs in W9706 were enriched in specific processes, including ABA signaling, wax biosynthesis, CHO metabolism, signal transduction and brassinosteroid biosynthesis-related processes, while the downregulated DEGs were enriched in specific processes, such as stomatal movement. Altogether, transcriptomic analysis suggests that the different drought resistances were correlated with the differential expression of genes, while the drought tolerance of W9706 is due to the more rapid response to stimulus, higher water retention capacity and stable cellular environment under water deficit conditions.

Novel flavin-containing monooxygenase protein FMO1 interacts with CAT2 to negatively regulate drought tolerance through ROS homeostasis and ABA signaling pathway in tomato.

Drought stress is the major abiotic factor that can seriously affect plant growth and crop production. The functions of flavin-containing monooxygenases (FMOs) are known in animals. They add molecular oxygen to lipophilic compounds or produce reactive oxygen species (ROS). However, little information on FMOs in plants is available. Here, we characterized a tomato drought-responsive gene that showed homology to FMO, and it was designated as FMO1. FMO1 was downregulated promptly by drought and ABA treatments. Transgenic functional analysis indicated that RNAi suppression of the expression of FMO1 (FMO1-Ri) improved drought tolerance relative to wild-type (WT) plants, whereas overexpression of FMO1 (FMO1-OE) reduced drought tolerance. The FMO1-Ri plants exhibited lower ABA accumulation, higher levels of antioxidant enzyme activities, and less ROS generation compared with the WT and FMO1-OE plants under drought stress. RNA-seq transcriptional analysis revealed the differential expression levels of many drought-responsive genes that were co-expressed with FMO1, including PP2Cs, PYLs, WRKY, and LEA. Using Y2H screening, we found that FMO1 physically interacted with catalase 2 (CAT2), which is an antioxidant enzyme and confers drought resistance. Our findings suggest that tomato FMO1 negatively regulates tomato drought tolerance in the ABA-dependent pathway and modulates ROS homeostasis by directly binding to SlCAT2.

Physiological and Transcriptomic Analyses Reveal the Response of Medicinal Plant Bletilla striata (Thunb. ex A. Murray) Rchb. f. via Regulating Genes Involved in the ABA Signaling Pathway, Photosynthesis, and ROS Scavenging under Drought Stress

Bletilla striata is a valuable Chinese herbal medicinal plant widely used in various fields. To meet the market demand for this herb, the tissue culture technology of B. striata was developed. However, drought stress has been a significant threat to the survival of cultivated B. striata. To further understand the underlying mechanisms of B. striata under drought stress, its response was investigated at the physiological and transcriptional levels. Our photosynthesis results show that the decline of the net photosynthesis rate (Pn) in B. striata leaves was mainly caused by nonstomatal limitation factors. Using transcriptomic analysis 2398, differentially expressed genes (DEGs) were identified. KEGG enrichment analysis showed that DEGs involved in plant hormone signal transduction (ko04075) were significantly altered, especially the abscisic-acid signaling pathway. The up-regulations of the serine/threonine protein kinase (SnRK2) and S-type anion (SLAH2) channels might lead to stomatal closure, which is the reason for decline of photosynthesis. Moreover, the downregulation of cytochrome b6 and photosystem I reaction center subunit III/IV might be the major reason for nonstomatal limitation. In addition, B. striata enhanced the ability of ROS scavenging via activating the gene expression of superoxide dismutase, catalase, and peroxidase in response to drought stress. Our study enhanced the understanding of B. striata in response to drought stress.

Transcriptional Plasticity and Cell Wall Characterization in High-Methanol-Producing Transgenic Tobacco Plants

Volatile organic compounds emanating from plant surfaces serve as a sustainable natural solution to combat biotic stresses in plants. Leaf methanol is the simplest and second major volatile organic compound after isoprene emitted through the leaf surface. Methanol has been neglected as a by-product of other secondary metabolites for a long time, but recent studies have suggested its importance in development and stress responses. In our previous findings, we had revealed that transgenic plants over-expressing PME, enhanced methanol production providing resistance against a broad spectrum of insects. In the current study, we extended our previous work to provide new insights by performing differential transcriptomics of high-methanol-producing insect-resistant transgenic lines. We found that 2262 genes were differentially expressed in the transgenics plants, including transcription factors, cell wall modulating, phytohormones signaling and development-related genes. Our results demonstrated that the expression levels of transcription factors associated with development and biotic stress were altered in the transgenic lines. In addition, phytohormones ABA and gibberellin signalling genes were upregulated, whereas ethylene and auxin signalling genes were downregulated. Moreover, biochemical characteristics of cell walls in both transgenic tobacco plants were comparable to the control plants.

Transcriptome and proteome depth analysis indicate ABA, MAPK cascade and Ca2+ signaling co-regulate cold tolerance in Rhododendron chrysanthum Pall.

Cold stress is a global common problem that significantly limits plant development and geographical distribution. Plants respond to low temperature stress by evolving interrelated regulatory pathways to respond and adapt to their environment in a timely manner. Rhodoendron chrysanthum Pall. (R. chrysanthum) is a perennial evergreen dwarf shrub used for adornment and medicine that thrives in the Changbai Mountains at high elevations and subfreezing conditions. In this study, a comprehensive investigation of cold tolerance (4°C, 12h) in R. chrysanthum leaves under cold using physiological combined with transcriptomic and proteomic approaches. There were 12,261 differentially expressed genes (DEGs) and 360 differentially expressed proteins (DEPs) in the low temperature (LT) and normal treatment (Control). Integrated transcriptomic and proteomic analyses showed that MAPK cascade, ABA biosynthesis and signaling, plant-pathogen interaction, linoleic acid metabolism and glycerophospholipid metabolism were significantly enriched in response to cold stress of R. chrysanthum leaves. We analyzed the involvement of ABA biosynthesis and signaling, MAPK cascade, and Ca2+ signaling, that may jointly respond to stomatal closure, chlorophyll degradation, and ROS homeostasis under low temperature stress. These results propose an integrated regulatory network of ABA, MAPK cascade and Ca2+ signaling comodulating the cold stress in R. chrysanthum, which will provide some insights to elucidate the molecular mechanisms of cold tolerance in plants.

Genome-wide identification of the ABA receptor PYL gene family and expression analysis in Prunus avium L.

The Pyrabactin resistance 1-like proteins (PYR/PYL/RCAR) are components of the ABA signaling pathway, playing a key role in how plants respond to abiotic stresses. Although PYL genes have been identified in a variety of plants, the evolution and structural characteristics of these genes in sweet cherry (Prunus avium L.) trees are largely unknown. In this study, 11 PaPYL genes were identified at genome-wide level in sweet cherry and were mapped to six chromosomes. According to the phylogenetic analysis, this gene family consisted of four subgroups; genes in the same subgroup had similar conserved motifs and the same number of exons, as revealed by analyzing gene structure. The intra-species collinearity analysis did not find any tandem duplication events in this gene family, and thus fragment duplication may be the primary reason for the generation of some PaPYL genes. Additionally, the inter-species collinearity analysis uncovered a potential evolutionary relationship between them. The expression analysis indicated that some PaPYL genes are specifically expressed in tissues, suggesting they may have unique functions. We also explored the expression of this gene family under different abiotic stress treatments (cold, salinity, drought). The qRT-PCR results showed that the expression levels of most genes were up-regulated under abiotic stress treatments, consistent with the results for their analyzed promoter cis-acting elements. This study provides a basis for further elucidating the function and expression patterning of the PYL gene family in sweet cherry, and helps to further analyze its regulatory mechanism under various abiotic stress factors.

Stem girdling enhances ABA-induced stomatal closure of phosphorus-deprived soybean plants

To investigate whether shoot-to-root communication modulates physiological (leaf water relations and growth), phytohormonal and transcriptomic responses to phosphorus (P) deprivation, soybean (Glycine max L. Merr.) plants received a factorial combination of low P conditions and girdling. Stem phloem tissue was surgically excised from the first internode between the cotyledonary and the first unifoliate leaf, immediately before imposing different P treatments (250 and 5 μM P in hydroponics). Root and leaf expression of genes involved in P homeostasis, sucrose metabolism and transporters, starch metabolism, hormone homeostasis and ABA signalling were analysed, to determine their regulation and relationships to tissue P / ABA concentrations. P deprivation mainly upregulated root and leaf P homeostasis gene expression only in intact plants, acting to maintain foliar P homeostasis. Although girdling suppressed root expression of these genes, it increased root P concentrations and decreased leaf P concentrations. Initially, P deprivation decreased stomatal conductance before leaf water status decreased, irrespective of girdling. Later, stomatal closure of girdled P-deprived plants was greater than intact P-deprived plants at the same, lower leaf water status, coincident with enhanced foliar ABA accumulation and expression of ABA biosynthesis genes. In contrast, P deprivation decreased foliar concentrations of the antitranspirant jasmonic acid and increased root and leaf SA concentrations at the end of the experiment. P deprivation also decreased foliar cytokinin and gibberellin concentrations before leaf growth was inhibited and leaf water status declined, suggesting non-hydraulic growth regulation. Girdling and P deprivation increased foliar sucrose and hexoses, corresponding to increased sucrose invertase and transporter gene expression in the leaves, while decreasing root sucrose and hexoses concentrations and downregulating sucrose metabolism but increasing transporter gene expression. Girdling perturbs multiple root and shoot phytohormonal responses that modulate physiological adaptation to P deprivation.

Transcriptome Identification of R2R3-MYB Gene Family Members in Pinus massoniana and PmMYB4 Response to Drought Stress

One of the largest families of transcription factors in plants, the MYB transcription factors family (Myeloblastosis, MYB TF), plays a vital role in regulating plant biochemical and physiological processes. The role of MYB TF in coping with stresses, such as drought, salt and cold, has been reported. Unfortunately, a comprehensive identification of R2R3-MYB TF in Masson pine (Pinus massoniana) has not been achieved. In this study, a total of 49 sequences were identified as R2R3-MYB TF. The structure, function and phylogenetic relationships of the conserved structural domains of Masson pine R2R3-MYB TF and Populus trichocarpa Torr. & A.Gray ex Hook. TFs were compared using bioinformatics tools. The results showed that Masson pine R2R3-MYB TF was divided into 24 groups, mainly located in the nucleus, and mostly lacking signal peptides and transmembrane structural domains with multiple phosphorylation sites. The drought stress-responsive R2R3-MYB gene, PmMYB4, was selected from the drought stress transcriptome based on analysis of the expression pattern and tissue specificity of PmMYB4 gene under abiotic stress using qPCR. The results showed that PmMYB4 can be involved in drought stress treatment through ABA signaling, as well as in multiple stress responses such as salt stress, and there were significant differences in the expression of PmMYB4 in the eight tissues. These results provide a reference scheme for the functional identification of R2R3-MYB transcription factors, which may be involved in plant responses to multiple stresses such as drought, and enrich our understanding of the functions of R2R3-MYB transcription factors in plants.

Overexpression of Water-Responsive Genes Promoted by Elevated CO2 Reduces ROS and Enhances Drought Tolerance in Coffea Species

Drought is a major constraint to plant growth and productivity worldwide and will aggravate as water availability becomes scarcer. Although elevated air [CO2] might mitigate some of these effects in plants, the mechanisms underlying the involved responses are poorly understood in woody economically important crops such as Coffea. This study analyzed transcriptome changes in Coffea canephora cv. CL153 and C. arabica cv. Icatu exposed to moderate (MWD) or severe water deficits (SWD) and grown under ambient (aCO2) or elevated (eCO2) air [CO2]. We found that changes in expression levels and regulatory pathways were barely affected by MWD, while the SWD condition led to a down-regulation of most differentially expressed genes (DEGs). eCO2 attenuated the impacts of drought in the transcripts of both genotypes but mostly in Icatu, in agreement with physiological and metabolic studies. A predominance of protective and reactive oxygen species (ROS)-scavenging-related genes, directly or indirectly associated with ABA signaling pathways, was found in Coffea responses, including genes involved in water deprivation and desiccation, such as protein phosphatases in Icatu, and aspartic proteases and dehydrins in CL153, whose expression was validated by qRT-PCR. The existence of a complex post-transcriptional regulatory mechanism appears to occur in Coffea explaining some apparent discrepancies between transcriptomic, proteomic, and physiological data in these genotypes.

Insights on the stem elongation of spur-type bud sport mutant of ‘Red Delicious’ apple

The decreased capacity of auxin-, CTK-, and BR-mediated cell division and cell enlargement pathways, combined with the enhanced capacity of GA and ETH-, JA-, ABA-, SA-mediated stress-resistant pathways were presumed to be the crucial reasons for the formation of spur-type 'Red Delicious' mutants. Vallee Spur', which exhibit short internodes and compact tree shape, is the fourth generation of the spur-type bud sport mutant of 'Red Delicious'. However, the underlying molecular mechanism of these properties remains unclear. Here, comparative phenotypic, full-length transcriptome and phytohormone analyses were performed between 'Red Delicious' (NSP) and 'Vallee Spur' (SP). The new shoot internode length of NSP was ˃ 1.53-fold higher than that of the SP mutant. Cytological analysis showed that the stem cells of the SP mutant were smaller and more tightly arranged relative to the NSP. By Iso-Seq, a total of 1426 differentially expressed genes (DEGs) were detected, including 808 upregulated and 618 downregulated genes in new shoot apex with 2 leaves of the SP mutant. Gene expressions involved in auxin, cytokinin (CTK), and brassinosteroid (BR) signal transduction were mostly downregulated in the SP mutant, whereas those involved in gibberellin (GA), ethylene (ETH), jasmonate (JA), ABA, and salicylic acid (SA) signal transduction were mostly upregulated. The overall thermogram analysis of hormone levels in the shoot apex carrying two leaves detected by LC-MS/MS absolute quantification showed that the levels of IAA-Asp, IAA, iP7G, OPDA, and 6-deoxyCS were significantly upregulated in the SP mutant, while the remaining 28 hormones were significantly downregulated. It is speculated that the decreased capacity of auxin, CTK, and BR-mediated cell division and cell enlargement pathways is crucial for the formation of the SP mutant. GA and stress-resistant pathways of ETH, JA, ABA, and SA also play vital roles in stem elongation. These results highlight the involvement of phytohormones in the formation of stem elongation occurring in 'Red Delicious' spur-type bud sport mutants and provide information for exploring its biological mechanism.