Post-germination Stages Research Articles

Effect of Different Basal Media and Organic Supplements on In Vitro Seedling Development of the Endangered Orchid Species Dendrobium moniliforme (L.) Swartz

The orchid Dendrobium moniliforme faces endangerment due to habitat loss and illegal harvesting, necessitating the development of an optimized artificial propagation system to aid conservation and reintroduction efforts. This study evaluated the effects of three plant growth media, namely Murashige and Skoog (MS), Hyponex, and Orchid Maintenance Medium (OMM) (P668), and various organic additives (apple homogenate, banana homogenate, and coconut water) on the in vitro seedling growth of D. moniliforme. The results reveal that, in early postgermination stages, seedlings achieve maximum growth in the Hyponex medium, with a fresh weight (92 mg) and root length (2.7 cm) approximately 20-fold greater than those in the MS medium and OMM. After 6 months, for seedlings grown in MS medium and OMM with banana (50 g·L−1), the mean fresh weights were 29 and 107 mg, respectively; however, the highest biomass was observed in seedlings grown in the Hyponex medium with coconut water (50 mL·L−1), exhibiting a mean fresh weight of 201 mg. This study highlights Hyponex medium with coconut water as the most effective combination for promoting D. moniliforme growth and identifies suitable organic supplements for the in vitro cultivation of seedlings from asymbiotic seed culture. This propagation system offers valuable technical support for the mass production and conservation of this epiphytic orchid.

Overexpression of histone demethylase gene SlJMJ18 and SlJMJ23 from tomato confers cadmium tolerance by regulating metal transport and hormone content in Arabidopsis

A lower concentration of cadmium (Cd), a hazardous and non-essential element for plant growth, will have deleterious effects on plants and endanger human health. Histone demethylase (JHDM) is important for plants' ability to withstand abiotic stress, according to an increasing number of studies. The degree of expression of the SlJMJ18 and SlJMJ23 genes in different tomato tissues was confirmed by this study. These two genes were responsive to the heavy metals Cd, Hg, Pb, and Cu stress, according to fluorescence quantification and GUS staining. Interestingly, the overexpression transgenic Arabidopsis plants of two genes have different responses to Cd stress. While SlJMJ18-OE lines consistently display Cd resistance but an early-flowering phenotype, SlJMJ23-OE plants have sensitivity during the post-germination stage and then greater tolerance to Cd stress. It was discovered that these two genes may affect cadmium tolerance of plants by regulating the expression of hormone synthesis related genes and hormone contents (BRs and ABA). Moreover, SlJMJ23 may resist cadmium stress by increasing the total phenol content in plants. The functional significance of JMJs is better understood in this study, which also offers a theoretical foundation for the use of molecular technology to develop plants resistant to Cd and an experimental basis for the efficient use of land resources.

Comparative proteomic analysis of seed germination between allotetraploid cotton Gossypium hirsutum and Gossypium barbadense

Seed germination, a key initial event in the plant life cycle, directly affects cotton yield and quality. Gossypium barbadense and Gossypium hirsutum gradually evolved through polyploidization, resulting in different characteristics, and this interspecific variation lacks genetic and molecular explanation. This work aimed to compare the proteomes between G. barbadense and G. hirsutum during seed germination. Here, we identified 2740 proteins for G. barbadense and 3758 for G. hirsutum. In the initial state, proteins in two cotton involved similar bioprocess, such as sugar metabolism, DNA repairing, and ABA signaling pathway. However, in the post-germination stage, G. hirsutum expressed more protein related to redox homeostasis, peroxidase activity, and pathogen interactions. Analyzing the different expression patterns of 915 single-copy orthogroups between the two kinds of cotton indicated that most of the differentially expressed proteins in G. barbadense were related to carbon metabolism. In contrast, most proteins in G. hirsutum were associated with stress response. Besides that, by proteogenomic analysis, we found 349 putative non-canonical peptides, which may be involved in plant development. These results will help to understand the different characteristics of these two kinds of cotton, such as fiber quality, yield, and adaptability. Significance statementCotton is the predominant natural fiber crop worldwide; Gossypium barbadense and Gossypium hirsutum have evolved through polyploidization to produce differing traits. However, given their specific features, the divergence of mechanisms underlying seed germination between G. hirsutum and G. barbadense has not been discussed. Here, we explore what protein contributes to interspecific differences between G. barbadense and G. hirsutum during the seed germination period. This study helps to elucidate the evolution and domestication history of cotton polyploids and may allow breeders to understand their domestication history better and improve fiber quality and adaptability.

Involvement of Abscisic Acid in Transition of Pea (Pisum sativum L.) Seeds from Germination to Post-Germination Stages.

The transition from seed to seedling represents a critical developmental step in the life cycle of higher plants, dramatically affecting plant ontogenesis and stress tolerance. The release from dormancy to acquiring germination ability is defined by a balance of phytohormones, with the substantial contribution of abscisic acid (ABA), which inhibits germination. We studied the embryonic axis of Pisum sativum L. before and after radicle protrusion. Our previous work compared RNA sequencing-based transcriptomics in the embryonic axis isolated before and after radicle protrusion. The current study aims to analyze ABA-dependent gene regulation during the transition of the embryonic axis from the germination to post-germination stages. First, we determined the levels of abscisates (ABA, phaseic acid, dihydrophaseic acid, and neo-phaseic acid) using ultra-high-performance liquid chromatography-tandem mass spectrometry. Second, we made a detailed annotation of ABA-associated genes using RNA sequencing-based transcriptome profiling. Finally, we analyzed the DNA methylation patterns in the promoters of the PsABI3, PsABI4, and PsABI5 genes. We showed that changes in the abscisate profile are characterized by the accumulation of ABA catabolites, and the ABA-related gene profile is accompanied by the upregulation of genes controlling seedling development and the downregulation of genes controlling water deprivation. The expression of ABI3, ABI4, and ABI5, which encode crucial transcription factors during late maturation, was downregulated by more than 20-fold, and their promoters exhibited high levels of methylation already at the late germination stage. Thus, although ABA remains important, other regulators seems to be involved in the transition from seed to seedling.

Morphophysiological indicators applied for the selection of different genotypes of Hevea spp. during germination and post-germination

We investigate and characterize the morphophysiological changes related to germination and the initial growth and development of seedlings of wild and cultivated genotypes of the genus Hevea spp. in finding to add value to new clones. Seeds from six genotypes ( H. brasiliensis (Willd. Ex A. Juss) Müll. Arg, H. guianensis Aubl., H. spruceana (Benth.) Müll. Arg., RRIM 600, Fx 3844, and IAN 873) were sterilized, and their imbibition curve, germination parameters, and initial seedling growth were characterized. In parallel, the morphology of the germination and post-germination stages was monitored from seeds germinated in washed sand. Morphological differences in the seeds/embryos, different strategies in the imbibition process, emergence, and initial seedling growth were observed. The species H. guianensis and H. spruceana showed the greatest differences in size, shape, and color of the seeds/embryos and initial seedling growth. The characteristics of the seed coat patterns, the seedling imbibition, growth, and development processes can serve as morphophysiological markers for genotype identification and selection. This is the first morphophysiological study of seeds and seedlings of wild and cultivated genotypes of species of the genus Hevea. We conclude that there is considerable phenotypic variability among species of the Hevea genus regarding seed and seedling morphophysiology, which can be explored in the rubber tree breeding programs.

Global Decrease in H3K9 Acetylation in Sorghum Seed Postgermination Stages.

Sorghum seed germination is accompanied by increases in nutrient contents and reduced levels of antinutrients and is therefore being applied to food processing. However, the characterization of acetylated histone H3 at lysine residue 9 (H3K9ac) in sorghum postgermination has lagged. In this study, we performed chromatin immunoprecipitation sequencing (ChIP-seq) to identify H3K9ac enrichment and obtained transcriptome in postgermination stages. More than 10,000 hypoacetylated genes gained H3K9ac marks in the postgermination stages. In addition, we observed that the expression of the main histone deacetylase (HDAC) genes was elevated. The application of the HDAC inhibitor trichostatin A (TSA) resulted in seed growth arrest, suggesting that the repression of the H3K9ac modification is critical for postgermination. Additionally, we obtained a comprehensive view of abundant genomic changes in H3K9ac-marked regions and transcription between the mock and TSA treatment groups, which suggested that H3K9ac was required in the late stage of autotrophic seedling establishment. Metabolic profiling, transcriptome analyses, and ChIP-seq revealed that H3K9ac is enriched at genes involved in phenylpropanoid, including lignin and flavonoid, biosynthesis. Our results suggest important roles of H3K9ac in sorghum seed postgermination stages.

ALLELOPATHIC EFFECT OF Chromolaena odorata FOLIAGE AS SOIL AMENDMENT ON THE GROWTH AND CHLOROPHYLL CONTENT OF Solanum lycopersicum, Mucuna pruriens, Abelmoschus esculentus and Citrullus lanatus.

This study investigated the allelopathic effect of Chromolaena odorata foliage used as soil amendments on the leaf area, plant height, number of leaves, plant biomass and total chlorophyll content of Solanum lycopersicum, Mucuna pruriens, Abelmuschus esculentus and Citrullus lanatus. Seeds of the test crops were planted in pots amended with 0%w/w, 1.9%w/w, 3.8%w/w, 5.7%w/w and 7.7%w/w of foliage of C. odorata and parameters as leaf area, plant height, number of leaves, plant biomass and total chlorophyll content were measured. The data collated were analysed using analysis of variance and the means separated using Tukey’s HSD. The leaf area, plant height, number of leaves and plant biomass showed a seemingly concentration-dependent, statistically significant increase in C. lanatus, S. lycopersicum and A. esculentus for all treatments, when compared to the 0%w/w treatment; and, although showing a visual and graphical increase for, M. pruriens, the increase in leaf area, plant height, number of leaves and plant biomass of M. pruriens were not statistically significant at 5% level of probability. The total chlorophyll content of all the plants studied showed concentration-dependent, statistically significant increments at 5% levels of probability, when compared to their respective 0%w/w treatments. The increase in the leaf area, plant height, number of leaves, plant biomass and total chlorophyll content of C. lanatus, S. lycopersicum, A. esculentus and M. pruriens due to the soil amendment with different amounts of C. odorata foliage is attributed to the possible effect of the allelochemicals in the leaves, which, although having a negative inhibitory effect on germination, had possibly undergone a bio-transformation to become stimulatory. Again, the foliar amendments increased the organic matter content of the soil, and thus, in synergy with the bio-transformed allelochemicals, serve as a bio-fertilizer to improve the growth of the test crops. Owing to the possible inhibitory effect of C. odorata foliage on the germination of these same test crops, it is therefore recommended that the foliage of C. odorata should only be used as organic manure or bio-fertilizer only at the post-germination stage of the test crops, not at the pre-germination stage.

The Aux/IAA protein TaIAA15-1A confers drought tolerance in Brachypodium by regulating abscisic acid signal pathway.

Overexpression of the Aux/IAA protein TaIAA15-1A from wheat improves drought tolerance by regulating the ABA signalling pathway in transgenic Brachypodium. Drought is a major abiotic stress that causes severe crop yield loss. Aux/IAA genes have been shown to be involved in drought stress responses. However, to the best of our knowledge, there has been little research on the molecular mechanism of the wheat Aux/IAA gene in the context of drought tolerance. In this study, we found that expression of the wheat Aux/IAA gene TaIAA15-1A was upregulated by PEG6000, NaCl, SA, JA, IAA and ABA. Transgenic plants overexpressing TaIAA15-1A showed higher drought tolerance than wild-type (WT) plants. The physiological analyses showed that the transgenic lines exhibited a higher survival rate, shoot length, and relative water content than the WT plants. The activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) were enhanced in transgenic lines, causing a reduction in the hydrogen peroxide (H2O2) and superoxide anion radical (O2-) contents. Transcriptome analysis showed that TaIAA15-1A overexpression alters the expression of these genes involved in the auxin signalling pathway, ABA signalling pathway, phenolamides and antioxidant pathways. The results of exogenous ABA treatment suggested that TaIAA15-1A overexpression increased sensitivity to ABA at the germination and postgermination stages compared to WT plants. These results indicate that TaIAA15-1A plays a positive role in plant drought tolerance by regulating ABA-related genes and improving antioxidative stress ability and has potential application in genetically modified crops.

SlGATA17, A tomato GATA protein, interacts with SlHY5 to modulate salinity tolerance and germination

The GATA transcription factor family participates in plant biological processes and abiotic stress across various crop species, but their function in response to salinity stress remains unexplored, particularly in tomato. Here, a tomato (Solanum lycopersicum) GATA gene, SlGATA17, was characterized in the tolerance to salinity stress. SlGATA17 had transcription activity in yeast, and the transcriptional activation domain was location on 32–60 amino acids of the N-terminal. SlGATA17 expression was induced by heat, cold, ABA, ACC and IAA treatments, but inhibited by NaCl and PEG treatments. In this study, the Slgata17 mutant plants showed slight injury symptoms compared to wild-type plants under salinity stress, and higher expression levels of stress-related marker genes (SlDREB2A, SlAREB1, SlRD29A and SlP5CS1). Heterologous expression of SlGATA17 in Arabidopsis promoted seed germination under NaCl, ABA and mannitol conditions, and inhibited ABA content and the expression levels of ABA related genes (AtNCED3, AtNCED5, AtNCED9, AtABI3, AtEM1 and AtEM6). However, SlGATA17 transgenic Arabidopsis were hypersensitive to NaCl at the post-germination stage. Yeast two assay and BiFC assay showed that tomato transcription factor ELONGATED HYPOCOTYL (SlHY5) interacts with SlGATA17. Yeast one assay and Dual-Luc assay showed that SlHY5 negatively regulated SlGATA17 expression via binding to the GATA-box of SlGATA17 promoter. SlHY5-SlGATA17 might function as a regulatory module in response to abiotic stress.

Chromatin dynamics associated with seed desiccation tolerance/sensitivity at early germination in Medicago truncatula.

Desiccation tolerance (DT) has contributed greatly to the adaptation of land plants to severe water-deficient conditions. DT is mostly observed in reproductive parts in flowering plants such as seeds. The seed DT is lost at early post germination stage but is temporally re-inducible in 1mm radicles during the so-called DT window following a PEG treatment before being permanently silenced in 5mm radicles of germinating seeds. The molecular mechanisms that activate/reactivate/silence DT in developing and germinating seeds have not yet been elucidated. Here, we analyzed chromatin dynamics related to re-inducibility of DT before and after the DT window at early germination in Medicago truncatula radicles to determine if DT-associated genes were transcriptionally regulated at the chromatin levels. Comparative transcriptome analysis of these radicles identified 948 genes as DT re-induction-related genes, positively correlated with DT re-induction. ATAC-Seq analyses revealed that the chromatin state of genomic regions containing these genes was clearly modulated by PEG treatment and affected by growth stages with opened chromatin in 1mm radicles with PEG (R1P); intermediate openness in 1mm radicles without PEG (R1); and condensed chromatin in 5mm radicles without PEG (R5). In contrast, we also showed that the 103 genes negatively correlated with the re-induction of DT did not show any transcriptional regulation at the chromatin level. Additionally, ChIP-Seq analyses for repressive marks H2AK119ub and H3K27me3 detected a prominent signal of H3K27me3 on the DT re-induction-related gene sequences at R5 but not in R1 and R1P. Moreover, no clear H2AK119ub marks was observed on the DT re-induction-related gene sequences at both developmental radicle stages, suggesting that silencing of DT process after germination will be mainly due to H3K27me3 marks by the action of the PRC2 complex, without involvement of PRC1 complex. The dynamic of chromatin changes associated with H3K27me3 were also confirmed on seed-specific genes encoding potential DT-related proteins such as LEAs, oleosins and transcriptional factors. However, several transcriptional factors did not show a clear link between their decrease of chromatin openness and H3K27me3 levels, suggesting that their accessibility may also be regulated by additional factors, such as other histone modifications. Finally, in order to make these comprehensive genome-wide analyses of transcript and chromatin dynamics useful to the scientific community working on early germination and DT, we generated a dedicated genome browser containing all these data and publicly available at https://iris.angers.inrae.fr/mtseedepiatlas/jbrowse/?data=Mtruncatula.

OsMas1, a novel maspardin protein gene, confers tolerance to salt and drought stresses by regulating ABA signaling in rice

Drought and salt stresses, the major environmental abiotic stresses in agriculture worldwide, affect plant growth, crop productivity, and quality. Therefore, developing crops with higher drought and salt tolerance is highly desirable. This study reported the isolation, biological function, and molecular characterization of a novel maspardin gene, OsMas1, from rice. The OsMas1 protein was localized to the cytoplasm. The expression levels of OsMas1 were up-regulated under mannitol, PEG6000, NaCl, and abscisic acid (ABA) treatments in rice. The OsMas1 gene was introduced into the rice cultivar Zhonghua 11 (wild type, WT). OsMas1-overexpression (OsMas1-OE) plants exhibited significantly enhanced salt and drought tolerance; in contrast, OsMas1-interference (OsMas1-RNAi) plants exhibited decreased tolerance to salt and drought stresses, compared with WT. OsMas1-OE plants exhibited enhanced hypersensitivity, while OsMas1-RNAi plants showed less sensitivity to exogenous ABA treatment at both germination and post-germination stages. ABA, proline and K+ contents and superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and photosynthesis activities were significantly increased. In contrast, malonaldehyde (MDA), hydrogen peroxide (H2O2), superoxide anion radical (O2-·), and Na+ contents were significantly decreased in OsMas1-OE plants compared with OsMas1-RNAi and WT plants. Overexpression of OsMas1 up-regulated the genes involved in ABA signaling, proline biosynthesis, reactive oxygen species (ROS)-scavenging system, photosynthesis, and ion transport under salt and drought stresses. Our results indicate that the OsMas1 gene improves salt and drought tolerance in rice, which may serve as a candidate gene for enhancing crop resistance to abiotic stresses.

Seed Transmission of Pathogens: Non-Canonical Immune Response in Arabidopsis Germinating Seeds Compared to Early Seedlings against the Necrotrophic Fungus Alternaria brassicicola.

The transmission of seed-borne pathogens by the germinating seed is responsible for major crop diseases. The immune responses of the seed facing biotic invaders are poorly documented so far. The Arabidopsis thaliana/Alternaria brassicicola patho-system was used to describe at the transcription level the responses of germinating seeds and young seedling stages to infection by the necrotrophic fungus. RNA-seq analyses of healthy versus inoculated seeds at 3 days after sowing (DAS), stage of radicle emergence, and at 6 and 10 DAS, two stages of seedling establishment, identified thousands of differentially expressed genes by Alternaria infection. Response to hypoxia, ethylene and indole pathways were found to be induced by Alternaria in the germinating seeds. However, surprisingly, the defense responses, namely the salicylic acid (SA) pathway, the response to reactive oxygen species (ROS), the endoplasmic reticulum-associated protein degradation (ERAD) and programmed cell death, were found to be strongly induced only during the latter post-germination stages. We propose that this non-canonical immune response in early germinating seeds compared to early seedling establishment was potentially due to the seed-to-seedling transition phase. Phenotypic analyses of about 14 mutants altered in the main defense pathways illustrated these specific defense responses. The unexpected germination deficiency and insensitivity to Alternaria in the glucosinolate deficient mutants allow hypothesis of a trade-off between seed germination, necrosis induction and Alternaria transmission to the seedling. The imbalance of the SA and jasmonic acid (JA) pathways to the detriment of the JA also illustrated a non-canonical immune response at the first stages of the seedling.

Seed-to-Seedling Transition in Pisum sativum L.: A Transcriptomic Approach.

The seed-to-seedling transition is a crucial step in the plant life cycle. The transition occurs at the end of seed germination and corresponds to the initiation of embryonic root growth. To improve our understanding of how a seed transforms into a seedling, we germinated the Pisum sativum L. seeds for 72 h and divided them into samples before and after radicle protrusion. Before radicle protrusion, seeds survived after drying and formed normally developed seedlings upon rehydration. Radicle protrusion increased the moisture content level in seed axes, and the accumulation of ROS first generated in the embryonic root and plumule. The water and oxidative status shift correlated with the desiccation tolerance loss. Then, we compared RNA sequencing-based transcriptomics in the embryonic axes isolated from pea seeds before and after radicle protrusion. We identified 24,184 differentially expressed genes during the transition to the post-germination stage. Among them, 2101 genes showed more prominent expression. They were related to primary and secondary metabolism, photosynthesis, biosynthesis of cell wall components, redox status, and responses to biotic stress. On the other hand, 415 genes showed significantly decreased expression, including the groups related to water deprivation (eight genes) and response to the ABA stimulus (fifteen genes). We assume that the water deprivation group, especially three genes also belonging to ABA stimulus (LTI65, LTP4, and HVA22E), may be crucial for the desiccation tolerance loss during a metabolic switch from seed to seedling. The latter is also accompanied by the suppression of ABA-related transcription factors ABI3, ABI4, and ABI5. Among them, HVA22E, ABI4, and ABI5 were highly conservative in functional domains and showed homologous sequences in different drought-tolerant species. These findings elaborate on the critical biochemical pathways and genes regulating seed-to-seedling transition.

Overexpression of histone demethylase gene SlJMJ524 from tomato confers Cd tolerance by regulating metal transport-related protein genes and flavonoid content in Arabidopsis

Cadmium (Cd), as a heavy metal, not only negatively affects the development and yield of plants, but also threatens human health due to its accumulation in plants. Increasing evidences indicate that the JUMONJI-C DOMAIN-CONTAINING PROTEIN (JMJ) gene family plays a key role in regulating plant development and stress. Therefore, in this study, SlJMJ524, a 1254 bp gene encoding the jumonji C domain (417 amino acids), was highly expressed in tomato leaves and flowers. Interestingly, the transgenic plants exhibited sensitivity to Cd during post-germination stage but showed enhanced tolerance to the heavy metal during adult stage. Overexpression of SlJMJ524 increased the expression level of related proteins gene involved in heavy metal uptake while increasing Cd tolerance through the GSH-PC pathway. The higher transcription of genes related to flavonoid synthesis reflected higher accumulations of flavonoids in transgenic plants. Our study demonstrated that the ectopic expression of SlJMJ524 conferred the transgenic plants many traits for improving cadmium stress tolerance at different developmental stages. This study advances our collective understanding of the functional role of JMJs and can be used to improve the cadmium tolerance and breeding of crops and plants.

Effect of drought stress on wheat (Triticum durum) growth and metabolism: insight from GABA shunt, reactive oxygen species and dehydrin genes expression.

Activation of γ-aminobutyric acid (GABA) shunt pathway and upregulation of dehydrins are involved in metabolic homeostasis and protective mechanisms against drought stress. Seed germination percentage, seedling growth, levels of GABA, alanine, glutamate, malondialdehyde (MDA), and the expression of glutamate decarboxylase (GAD ) and dehydrin (dhn and wcor ) genes were examined in post-germination and seedlings of four durum wheat (Triticum durum L.) cultivars in response to water holding capacity levels (80%, 50%, and 20%). Data showed a significant decrease in seed germination percentage, seedling length, fresh and dry weight, and water content as water holding capacity level was decreased. Levels of GABA, alanine, glutamate, and MDA were significantly increased with a negative correlation in post-germination and seedling stages as water holding capacity level was decreased. Prolonged exposure to drought stress increased the GAD expression that activated GABA shunt pathway especially at seedlings growth stage to maintain carbon/nitrogen balance, amino acids and carbohydrates metabolism, and plant growth regulation under drought stress. The mRNA transcripts of dhn and wcor significantly increased as water availability decreased in all wheat cultivars during the post-germination stage presumably to enhance plant tolerance to drought stress by cell membrane protection, cryoprotection of enzymes, and prevention of reactive oxygen species (ROS) accumulation. This study showed that the four durum wheat cultivars responded differently to drought stress especially during the seedling growth stage which might be connected with ROS scavenging systems and the activation of antioxidant enzymes that were associated with activation of GABA shunt pathway and the production of GABA in durum seedlings.

OsIAA18, an Aux/IAA Transcription Factor Gene, Is Involved in Salt and Drought Tolerance in Rice.

Auxin/indoleacetic acid (Aux/IAA) proteins play an important regulatory role in the developmental process of plants and their responses to stresses. A previous study has shown that constitutive expression of OsIAA18, an Aux/IAA transcription factor gene of rice improved salt and osmotic tolerance in transgenic Arabidopsis plants. However, little work is known about the regulatory functions of the OsIAA18 gene in regulating the abiotic stress tolerance of rice. In this study, the OsIAA18 gene was introduced into the rice cultivar, Zhonghua 11 and the OsIAA18 overexpression in rice plants exhibited significantly enhanced salt and drought tolerance compared to the wild type (WT). Moreover, overexpression of OsIAA18 in rice increased endogenous levels of abscisic acid (ABA) and the overexpression of OsIAA18 in rice plants showed hypersensitivity to exogenous ABA treatment at both the germination and postgermination stages compared to WT. Overexpression of OsIAA18 upregulated the genes involved in ABA biosynthesis and signaling pathways, proline biosynthesis pathway, and reactive oxygen species (ROS)-scavenging system in the overexpression of OsIAA18 in rice plants under salt and drought stresses. Proline content, superoxide dismutase (SOD), and peroxidase (POD) activities were significantly increased, whereas malonaldehyde (MDA), hydrogen peroxide (H2O2), and superoxide anion radical (O2–) content were significantly decreased in the transgenic plants under salt and drought stresses. Taken together, we suggest that OsIAA18 plays a positive role in drought and salt tolerance by regulating stress-induced ABA signaling. The OsIAA18 gene has a potential application in genetically modified crops with enhanced tolerance to abiotic stresses.

Differential Physiological Responses to Salt Stress between Salt-Sensitive and Salt-Tolerant japonica Rice Cultivars at the Post-Germination and Seedling Stages.

Soil salinity is a key source of abiotic stress in the cultivation of rice. In this study, two currently cultivated japonica rice species—Zhegeng 78 (salt-tolerant) and Zhegeng 99 (salt-sensitive)—with similar backgrounds were identified and used to investigate their differential responses to salt stress at the post-germination and seedling stages. Quantitative RT-PCR analysis demonstrated that the expression of OsSOS1, OsHAK1, and OsHAK5 at the post-germination stage, and the expression of OsHKT1,1, OsHTK2,1, and OsHAK1 at the seedling stage, were significantly higher in the salt-tolerant Zhegeng 78 compared with those of the salt-sensitive Zhegeng 99 under salt stress. The significantly lower Na+ net uptake rate at the post-germination and higher K+ net uptake rates at the post-germination and seedling stages were observed in the salt-tolerant Zhegeng 78 compared with those of the salt-sensitive Zhegeng 99 under salt stress. Significantly higher activity of peroxidase (POD) and the lower hydrogen peroxide (H2O2) accumulation were observed in the salt-tolerant Zhegeng 78 compared with those of salt-sensitive Zhegeng 99 under salt stress at the seeding stage. The salt-tolerant Zhegeng 78 might be valuable in future cultivation in salinity soils.

Dynamic Transcriptomic and Metabolomic Analyses of Madhuca pasquieri (Dubard) H. J. Lam During the Post-germination Stages.

The wild population of Madhuca pasquieri (Dubard) H. J. Lam is currently dwindling; its understory seedlings are rare, and there is a lack of molecular studies, which impedes the conservation of this species. This study exploited second-generation sequencing and widely targeted metabolomics analysis to uncover the dynamic changes in differentially expressed genes (DEGs) and differentially accumulated metabolites (DAMs) in five post-germination stages of M. pasquieri whole organism. Notably, the weighted gene co-expression network analysis (WGCNA), transcriptome, and metabolome association analyses all indicated significant enrichment of the flavonoid biosynthesis pathway in stage 4 (two-leaf), and an upregulation of the genes encoding flavonol biosynthesis in this stage. In stage 5 (nine-leaf), the flavonols were significantly accumulated, indicating that the changes in metabolites were driven at the transcript level. According to the significant changes in gene expression encoding auxin transport carriers and their correlation with flavonols during stage 5, the flavonols were speculated to have a direct inhibitory effect on the expression of PIN4 encoding gene, which may inhibit the process of polar auxin transport. The results provided important insights into the molecular network relationships between the transcription and metabolism of this rare and endangered species during the post-germination stages and explained the reasons for the slow growth of its seedlings at the molecular level.

The miR169n-NF-YA8 regulation module involved in drought resistance in Brassica napus L

As an ancient and conserved plant microRNA (miRNA) family, miR169 targets nuclear factor Y subunit alpha (NF-YA) family members. The miR169-NF-YA module is associated with plant development and various stress responses. However, the function of miR169 in response to drought stress in rapeseed (Brassica napus L.) is unclear. In the present study, we showed that miR169n acted as a negative regulator of drought resistance in rapeseed by targeting a nuclear factor Y-A gene, NF-YA8. miR169n was strongly down-regulated by drought stress. Expression of a miR169n target mimicry construct (MIM169n) which functioned as a sponge to trap miR169n resulted in enhanced resistance of transgenic plants to both osmotic stress at the post-germination stage and drought stress at the seedling stage. MIM169n plants had a higher relative water content (RWC) and proline content, lower relative electrolyte leakage (REL), and showed higher antioxidative capability compared with those of control (CK) plants under drought stress. Moreover, NF-YA8 was verified as a target of miR169n, and overexpression of NF-YA8 led to improved tolerance of rapeseed to osmotic stress at the post-germination stage. Overall, our findings implied that the miR169n-NF-YA8 regulatory module could serve as a potential target for genetic improvement of drought resistance in B. napus.

Overexpression of OsUGT3 enhances drought and salt tolerance through modulating ABA synthesis and scavenging ROS in rice

Abscisic acid (ABA) is an important plant hormone that plays a crucial role in response to abiotic stresses. It is well-known that the level of ABA will be dynamically regulated under stress conditions, however, the regulatory mechanism of ABA homeostasis remains largely unknown. In this study, we isolated and characterized a putative UDP-glycosyltransferase (UGT3) in rice, which is proven to be stress-induced and involved in ABA dynamic change. Our data demonstrated that overexpression of the glycosyltransferase gene UGT3 enhanced drought and salt stress tolerance of the transgenic rice. However, ugt3 mutant exhibited sensitive phenotypes under stresses. The ectopic expression of UGT3 in Arabidopsis also enhanced the plant tolerance to drought and salt stresses. Overexpression of UGT3 in rice increased endogenous ABA level and showed hypersensitive phenotype to exogenous ABA treatment at both germination and post-germination stages. Further studies demonstrated that UGT3 elevated activities of the antioxidant enzymes and reduced the production of reactive oxygen species to control the oxidative burst under stress conditions. Taken together, this work reveals that glycosyltransferase UGT3 can cope with the environmental challenges through a possible interactive network of ABA, ROS and antioxidants under abiotic stresses.