BrPARP1, a Poly (ADP-Ribose) Polymerase Gene, Is Involved in Root Development in Brassica rapa under Drought Stress

PARP proteins are highly conserved homologs among the eukaryotic poly (ADP-ribose) polymerases. After activation, ADP-ribose polymers are synthesized on a series of ribozymes that use NAD+ as a substrate. PARPs participate in the regulation of various important biological processes, such as plant growth, development, and stress response. In this study, we characterized the homologue of PARP1 in B. rapa using RNA interference (RNAi) to reveal the underlying mechanism responding to drought stress. Bioinformatics and expression pattern analyses demonstrated that two copy numbers of PARP1 genes (BrPARP1.A03 and BrPARP1.A05) in B. rapa following a whole-genome triplication (WGT) event were retained compared with Arabidopsis, but only BrPARP1.A03 was predominantly transcribed in plant roots. Silencing of BrPARP1 could markedly promote root growth and development, probably via regulating cell division, and the transgenic Brassica lines showed more tolerance under drought treatment, accompanied with substantial alterations including accumulated proline contents, significantly reduced malondialdehyde, and increased antioxidative enzyme activity. In addition, the findings showed that the expression of stress-responsive genes, as well as reactive oxygen species (ROS)-scavenging related genes, was largely reinforced in the transgenic lines under drought stress. In general, these results indicated that BrPARP1 likely responds to drought stress by regulating root growth and the expression of stress-related genes to cope with adverse conditions in B. rapa.

The Rice Abscisic Acid-Responsive RING Finger E3 Ligase OsRF1 Targets OsPP2C09 for Degradation and Confers Drought and Salinity Tolerance in Rice

Drought and salinity are major important factors that restrain growth and productivity of rice. In plants, many really interesting new gene (RING) finger proteins have been reported to enhance drought and salt tolerance. However, their mode of action and interacting substrates are largely unknown. Here, we identified a new small RING-H2 type E3 ligase OsRF1, which is involved in the ABA and stress responses of rice. OsRF1 transcripts were highly induced by ABA, salt, or drought treatment. Upregulation of OsRF1 in transgenic rice conferred drought and salt tolerance and increased endogenous ABA levels. Consistent with this, faster transcriptional activation of key ABA biosynthetic genes, ZEP, NCED3, and ABA4, was observed in OsRF1-OE plants compared with wild type in response to drought stress. Yeast two-hybrid assay, BiFC, and co-immunoprecipitation analysis identified clade A PP2C proteins as direct interacting partners with OsRF1. In vitro ubiquitination assay indicated that OsRF1 exhibited E3 ligase activity, and that it targeted OsPP2C09 protein for ubiquitination and degradation. Cell-free degradation assay further showed that the OsPP2C09 protein is more rapidly degraded by ABA in the OsRF1-OE rice than in the wild type. The combined results suggested that OsRF1 is a positive player of stress responses by modulating protein stability of clade A PP2C proteins, negative regulators of ABA signaling.

Trophic niche but not abundance of Collembola and Oribatida changes with drought and farming system

Higher frequencies of summer droughts are predicted to change soil conditions in the future affecting soil fauna communities and their biotic interactions. In agroecosystems drought effects on soil biota may be modulated by different management practices that alter the availability of different food resources. Recent studies on the effect of drought on soil microarthropods focused on measures of abundance and diversity. We here additionally investigated shifts in trophic niches of Collembola and Oribatida as indicated by stable isotope analysis (13C and 15N). We simulated short-term summer drought by excluding 65% of the ambient precipitation in conventionally and organically managed winter wheat fields on the DOK trial in Switzerland. Stable isotope values suggest that plant litter and root exudates were the most important resources for Collembola (Isotoma caerulea, Isotomurus maculatus and Orchesella villosa) and older plant material and microorganisms for Oribatida (Scheloribates laevigatus and Tectocepheus sarekensis). Drought treatment and farming systems did not affect abundances of the studied species. However, isotope values of some species increased in organically managed fields indicating a higher proportion of microorganisms in their diet. Trophic niche size, a measure of both isotope values combined, decreased with drought and under organic farming in some species presumably due to favored use of plants as basal resource instead of algae and microorganisms. Overall, our results suggest that the flexible usage of resources may buffer effects of drought and management practices on the abundance of microarthropods in agricultural systems.

Profiling of N6-Methyladenosine (m6A) Modification Landscape in Response to Drought Stress in Apple (Malus prunifolia (Willd.) Borkh)

Drought stress is a significant environmental factor limiting crop growth worldwide. Malus prunifolia is an important apple species endemic to China and is used for apple cultivars and rootstocks with great drought tolerance. N6-methyladenosine (m6A) is a common epigenetic modification on messenger RNAs (mRNAs) in eukaryotes which is critical for various biological processes. However, there are no reports on m6A methylation in apple response to drought stress. Here, we assessed the m6A landscape of M. prunifolia seedlings in response to drought and analyzed the association between m6A modification and transcript expression. In total, we found 19,783 and 19,609 significant m6A peaks in the control and drought treatment groups, respectively, and discovered a UGUAH (H: A/U/C) motif. In M. prunifolia, under both control and drought conditions, peaks were highly enriched in the 3′ untranslated region (UTR) and coding sequence (CDS). Among 4204 significant differential m6A peaks in drought-treated M. prunifolia compared to control-treated M. prunifolia, 4158 genes with m6A modification were identified. Interestingly, a large number of hypermethylated peaks (4069) were stimulated by drought treatment compared to hypomethylation. Among the hypermethylated peak-related genes, 972 and 1238 differentially expressed genes (DEGs) were up- and down-regulated in response to drought, respectively. Gene ontology (GO) analyses of differential m6A-modified genes revealed that GO slims related to RNA processing, epigenetic regulation, and stress tolerance were significantly enriched. The m6A modification landscape depicted in this study sheds light on the epigenetic regulation of M. prunifolia in response to drought stress and indicates new directions for the breeding of drought-tolerant apple trees.

Growth-limiting drought stress induces time-of-day dependent transcriptome and physiological responses in hybrid poplar

Drought stress negatively impacts the health of long-lived trees. Understanding the genetic mechanisms that underpin response to drought stress is requisite for selecting or enhancing climate change resilience. We aimed to determine how hybrid poplars respond to prolonged and uniform exposure to drought; how responses to moderate and more severe growth-limiting drought stresses differed; and, how drought responses change throughout the day. We established hybrid poplar trees (Populus x 'Okanese') from unrooted stem cutting with abundant soil moisture for six weeks. We then withheld water to establish well-watered, moderate, and severe growth-limiting drought conditions. These conditions were maintained for three weeks during which growth was monitored. We then measured photosynthetic rates and transcriptomes of leaves that had developed during the drought treatments at two times of day. The moderate and severe drought treatments elicited distinct changes in growth and development, photosynthetic rates, and global transcriptome profiles. Notably, the time of day of sampling produced the strongest signal in the transcriptome data. The moderate drought treatment elicited global transcriptome changes that were intermediate to the severe and well-watered treatments in the early evening, but did not elicit a strong drought response in the morning, emphasizing the complex nature of drought regulation in long-lived trees.

Transcriptome Reveals the Dynamic Response Mechanism of Pearl Millet Roots under Drought Stress

Drought is a major threat to global agricultural production that limits the growth, development and survival rate of plants, leading to tremendous losses in yield. Pearl millet (Cenchrus americanus (L.) Morrone) has an excellent drought tolerance, and is an ideal plant material for studying the drought resistance of cereal crops. The roots are crucial organs of plants that experience drought stress, and the roots can sense and respond to such conditions. In this study, we explored the mechanism of drought tolerance of pearl millet by comparing transcriptomic data under normal conditions and drought treatment at four time points (24 h, 48 h, 96 h, and 144 h) in the roots during the seedling stage. A total of 1297, 2814, 7401, and 14,480 differentially expressed genes (DEGs) were found at 24 h, 48 h, 96 h, and 144 h, respectively. Based on Kyoto Encyclopedia of Genes and Genomes and Gene Ontology enrichment analyses, we found that many DEGs participated in plant hormone-related signaling pathways and the “oxidoreductase activity” pathway. These results should provide a theoretical basis to enhance drought resistance in other plant species.

Functional Characterization of Serotonin N-Acetyltransferase Genes (SNAT1/2) in Melatonin Biosynthesis of Hypericum perforatum

Hypericum perforatum is a traditional medicinal plant that contains various secondary metabolites. As an active component in H. perforatum, melatonin plays important role in plant antioxidation, growth, and photoperiod regulation. Serotonin N-acetyltransferase (SNAT) is the key enzyme involved in the last or penultimate step of phytomelatonin biosynthesis. A total of 48 members of SNAT family were screened and analyzed based on the whole genome data of H. perforatum, and two SNAT genes (HpSNAT1 and HpSNAT2) were functionally verified to be involved in the biosynthesis of melatonin. It was found that HpSNAT1 and HpSNAT2 were highly expressed in the leaves and showed obvious responses to high salt and drought treatment. Subcellular localization analysis indicated that these two proteins were both localized in the chloroplasts by the Arabidopsis protoplasts transient transfection. Overexpression of HpSNAT1 and HpSNAT2 in Arabidopsis (SNAT) and H. perforatum (wild-type) resulted in melatonin content 1.9–2.2-fold and 2.5–4.2-fold higher than that in control groups, respectively. Meanwhile, SNAT-overexpressing Arabidopsis plants showed a stronger ability of root growth and scavenging endogenous reactive oxygen species. In this study, the complete transgenic plants of H. perforatum were obtained through Agrobacterium-mediated genetic transformation for the first time, which laid a significant foundation for further research on the function of key genes in H. perforatum.

The Physiological Responses of Zea Mays L. and Cucumis Sativus L. on Drought Stress and Re-Watering

Drought leads to deficit water availability and its detrimental effects seriously threaten plant growth. This study assessed the physiological, biochemical, and antioxidant adjustments in different types of photosynthetic plants between Zea mays L. (C4) and Cucumis sativus L. (C3 plant) under response to short-term drought stress. Analyses of relative water content (RWC), proline, and ascorbic acid (AsA) were performed to explore how these plants react to drought. Fifteen-day-old plants were subjected to full irrigation or gradual drought periods for 2-d, 4-d, 6-d, and 8-d following by recovery for 7-d. The results revealed that drought significantly reduces leaf RCW in both plants. Re-watered Z. mays after 8-d drought was higher than C. sativus and reestablished RCW by 23% of stressed plant although remained lower by 9% of the well-watered plant. While, proline and AsA contents in Z. mays were higher than those in C. sativus in drought treatment at 8-d (2.05 µmol/g FW) and 6-d (3174.60 AsA/100 g FW), respectively, that could demonstrate osmotic adjustment ability in this C4 species. The increased proline in both plants also indicates a good strategy for plants to recover. Rewatering gave a decrease AsA and could be expected that plants restore cellular activity after oxidative injury. Based on our study, proline is the most informative biochemical marker to differentiate plant response to drought and Z. mays adjusted defense mechanism to drought rather than C. sativus due to higher accumulation of proline, better antioxidant activity, and improved RCW after recovery.

Contents of lobetyolin, syringin, and atractylolide III in Codonopsis pilosula are related to dynamic changes of endophytes under drought stress

Background Codonopsis pilosula, an important medicinal plant, can accumulate certain metabolites under moderate drought stress. Endophytes are involved in the metabolite accumulations within medicinal plants. It is still unknown that the endophytes of C. pilosula are associated with the accumulations of metabolites. This study aims to investigate the promoting effect of endophytes on the accumulations of active substances in C. pilosula under drought stress. Methods High–performance liquid chromatography and high–throughput sequencing technology were performed to investigate changes in the contents of secondary metabolite and endophyte abundances of C. pilosula under drought stress, respectively. Spearman’s correlation analysis was further conducted to identify the endophytic biomarkers related to accumulations of pharmacodynamic compounds. Culture-dependent experiments were performed to confirm the functions of endophytes in metabolite accumulations. Results The distribution of pharmacological components and diversity and composition of endophytes showed tissue specificity within C. pilosula. The contents of lobetyolin, syringin, and atractylolide III in C. pilosula under drought stress were increased by 8.47%‒86.47%, 28.78%‒230.98%, and 32.17%‒177.86%, respectively, in comparison with those in untreated groups. The Chao 1 and Shannon indices in different parts of drought–stressed C. pilosula increased compared with those in untreated parts. The composition of endophytic communities in drought treatment parts of C. pilosula was different from that in control parts. A total of 226 microbial taxa were identified as potential biomarkers, of which the abundances of 42 taxa were significantly and positively correlated to the pharmacodynamic contents. Culture-dependent experiments confirmed that the contents of lobetyolin and atractylolide III were increased by the application of Epicoccum thailandicum, Filobasidium magnum, and Paraphoma rhaphiolepidis at the rates of 11.12%‒46.02%, and that the content of syringin was increased by Pseudomonas nitroreducens at the rates of 118.61%‒119.36%. Conclusions Certain endophytes participated in the accumulations of bioactive metabolites, which provided a scientific evidence for the development and application of microorganisms to improve the quality of traditional Chinese medicine.