Flooding Stress Response Research Articles

Phytohormone and energy metabolism of rice germination and coleoptile growth during submergence by RNA-seq and LC-MS/MS analysis

Rice is one of the main food crops to guarantee the world's food security. Although rice is a semi-aquatic crop, long-term flooding stress can also inhibit growth and reduce yield and quality. Especially during the germination period, the germination rate of rice was inhibited by flooding stress, which resulted in the low emergence rate. Therefore, in this study, rice lines or cultivars with flood tolerance (C11) and flood sensitivity (KY60) were screened, and analyzed the response mechanisms during seed germination at different time points of waterlogging stress through transcriptome and metabolome sequencing. Compared with C11, waterlogging stress significantly inhibited the germination and coleoptile growth of KY60. Certain pathways, including plant hormones, energy metabolism, and carbohydrate metabolism, were identified as significantly involved in the flooding stress response in rice. The plant hormone-related DEGs (IAA, SAUR, ARF, LOX) and metabolites (IAA, JA, GA, and SA) were identified to up-regulate gene expression levels and significantly up-accumulate phytohormone content in C11 to resist waterlogging stress. However, the ABA signaling pathway-related genes (ZHD, PLY) and ABA content were decreased in C11 under submerged conditions. It is beneficial for C11 to promote seed germination and coleoptile elongation under flooding conditions. A total of 150 DEGs (AMY, PGLR) and 58 related compounds were identified as involved in the energy metabolism and carbohydrate metabolism of rice after flooding stress. Candidate key genes and metabolite compounds were identified by combined RNA-Seq and LC-MS/MS correlation network analysis. This study provided insights to explore the flood-tolerance mechanism of rice and laid the foundation for the cultivation of new varieties of flood-tolerance rice.

'Against all floods': plant adaptation to flooding stress and combined abiotic stresses.

Current climate change brings with it a higher frequency of environmental stresses, which occur in combination rather than individually leading to massive crop losses worldwide. In addition to, for example, drought stress (low water availability), also flooding (excessive water) can threaten the plant, causing, among others, an energy crisis due to hypoxia, which is responded to by extensive transcriptional, metabolic and growth-related adaptations. While signalling during flooding is relatively well understood, at least in model plants, the molecular mechanisms of combinatorial flooding stress responses, for example, flooding simultaneously with salinity, temperature stress and heavy metal stress or sequentially with drought stress, remain elusive. This represents a significant gap in knowledge due to the fact that dually stressed plants often show unique responses at multiple levels not observed under single stress. In this review, we (i) consider possible effects of stress combinations from a theoretical point of view, (ii) summarize the current state of knowledge on signal transduction under single flooding stress, (iii) describe plant adaptation responses to flooding stress combined with four other abiotic stresses and (iv) propose molecular components of combinatorial flooding (hypoxia) stress adaptation based on their reported dual roles in multiple stresses. This way, more future emphasis may be placed on deciphering molecular mechanisms of combinatorial flooding stress adaptation, thereby potentially stimulating development of molecular tools to improve plant resilience towards multi-stress scenarios.

A balancing act: ERFVII feedback loops finetune the flooding stress response.

A balancing act: ERFVII feedback loops finetune the flooding stress response.

Comparative transcriptome profiling of two sweetpotato cultivars with contrasting flooding stress tolerance levels

Sweetpotato (Ipomoea batatas [L.] Lam) is an important starch crop that ensures food and nutrition security in the era of climate change. Sweetpotato is tolerant to environmental stresses such as drought, high temperature, and high salt, and therefore, is well adapted to marginal lands; however, it is relatively vulnerable to flooding stress, which severely reduces its yield and commercial value. To understand the flooding stress response of sweetpotato, we performed comparative transcriptome analysis of the leaves of two sweetpotato cultivars with contrasting flooding stress tolerance levels: Yeonjami (YJM; flooding tolerant) and Jeonmi (JM; flooding sensitive). Both cultivars were partially submerged in water for 0, 0.5, and 3 days. RNA-seq data of both cultivars revealed 14,229 differentially expressed genes (DEGs), which were categorized into seven clusters and six groups, based on the expression pattern of co-expressed DEGs and expression duration of DEGs, respectively. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses showed that DEGs of distinguishing functions between the two cultivars were involved in plant hormone signaling, carbohydrate transport, and mitogen-activated protein kinase (MAPK) signaling. Based on these results, we predict that YJM promotes adventitious growth, whereas JM exhibits shoot elongation under flooding stress. The expression levels of several key candidate genes involved in flooding tolerance correlated well with the comparative transcriptomics data. Overall, this study provides further insights into the molecular mechanism of flooding stress response in sweetpotato, and reveals candidate genes that could be used for developing new flooding tolerant sweetpotato cultivars.

Day-Length Is Involved in Flooding Tolerance Response in Wild Type and Variant Genotypes of Rootstock Prunus cerasifera L.

Current and predicted climate changes scenarios require crops with an improved adaptability to mutable environmental features, such as, hypoxia for the root system. In order to overcome the reduction of oxygen, plants activate coping mechanisms and strategies. Prunus spp. are hypoxia-sensitive woody species and although many information has been gathered over the last decades, many physiological mechanisms remain unclear. To verify whether anoxic plant responses are also regulated by photoperiod, plants of Mr.S.2/5-WT plum, and its variant genotypes S.4 tolerant (plus) and S.1 sensitive (minus) to flooding, were grown in a greenhouse and were submitted to natural photoperiod (NP) and to constant photoperiod (CP) from mid-July until the first 10 days of October. From mid-September plants from each genotype, grown under the two photoperiods, were divided into two groups, and one of them underwent long-term flooding. Gas exchange parameters, energetic and biochemical activities, leaf chlorophyll contents, and stress symptoms were measured at different times, whereas soluble sugars were quantified in leaves and roots 14 days after flooding, when stress symptoms in WT and S.1 became prominent. Seasonal changes in the photoperiod played a role in the adaptability to anoxia, although flooding stress response differed among the three genotypes. Anoxia affected leaf gas exchange and S.4 flooded-leaves retained higher ACO2 under conditions of NP and CP. Leaf soluble sugar concentration differed among genotypes. Regardless the photoperiod, S.4 anoxic-leaf sugar concentration was the lowest, except for sorbitol. S.4 anoxic-roots under CP accumulated the highest levels of sucrose and sorbitol. Influences of the photoperiod were observed in WT and S.1 anoxic-leaves, whereas S.1 anoxic roots accumulated the lowest concentration of sugars, regardless of photoperiod. Leaf and root respiratory activity in flooded-plants was highest in S.4, and ADH activity increased in all flooded plants under CP but the highest activity was observed only in S.1 under NP during flooding. Results are consistent with the hypothesis that the S.4 genotype has a plastic adaptability to flooding stress, escaping from the photoperiod regulatory cross-talk system, and can better cope with the new scenarios generated by climate changes.

Characterization of the XTH Gene Family: New Insight to the Roles in Soybean Flooding Tolerance.

Xyloglucan endotransglycosylases/hydrolases (XTHs) are a class of enzymes involved in the construction and remodeling of cellulose/xyloglucan crosslinks and play an important role in regulating cell wall extensibility. However, little is known about this class of enzymes in soybean. Here, 61 soybean XTH genes (GmXTHs) were identified and classified into three subgroups through comparative phylogenetic analysis. Genome duplication greatly contributed to the expansion of GmXTH genes in soybean. A conserved amino acid motif responsible for the catalytic activity was identified in all GmXTHs. Further expression analysis revealed that most GmXTHs exhibited a distinct organ-specific expression pattern, and the expression level of many GmXTH genes was significantly associated with ethylene and flooding stress. To illustrate a possible role of XTH genes in regulating stress responses, the Arabidopsis AtXTH31 gene was overexpressed in soybean. The generated transgenic plants exhibited improved tolerance to flooding stress, with a higher germination rate and longer roots/hypocotyls during the seedling stage and vegetative growth stages. In summary, our combined bioinformatics and gene expression pattern analyses suggest that GmXTH genes play a role in regulating soybean stress responses. The enhanced soybean flooding tolerance resulting from the expression of an Arabidopsis XTH also supports the role of XTH genes in regulating plant flooding stress responses.

Leaf transcriptome of two highly divergent genotypes of Urochloa humidicola (Poaceae), a tropical polyploid forage grass adapted to acidic soils and temporary flooding areas.

Background Urochloa humidicola (Koronivia grass) is a polyploid (6x to 9x) species that is used as forage in the tropics. Facultative apospory apomixis is present in most of the genotypes of this species, although one individual has been described as sexual. Molecular studies have been restricted to molecular marker approaches for genetic diversity estimations and linkage map construction. The objectives of the present study were to describe and compare the leaf transcriptome of two important genotypes that are highly divergent in terms of their phenotypes and reproduction modes: the sexual BH031 and the aposporous apomictic cultivar BRS Tupi.ResultsWe sequenced the leaf transcriptome of Koronivia grass using an Illumina GAIIx system, which produced 13.09 Gb of data that consisted of 163,575,526 paired-end reads between the two libraries. We de novo-assembled 76,196 transcripts with an average length of 1,152 bp and filtered 35,093 non-redundant unigenes. A similarity search against the non-redundant National Center of Biotechnology Information (NCBI) protein database returned 65 % hits. We annotated 24,133 unigenes in the Phytozome database and 14,082 unigenes in the UniProtKB/Swiss-Prot database, assigned 108,334 gene ontology terms to 17,255 unigenes and identified 5,324 unigenes in 327 known metabolic pathways. Comparisons with other grasses via a reciprocal BLAST search revealed a larger number of orthologous genes for the Panicum species. The unigenes were involved in C4 photosynthesis, lignocellulose biosynthesis and flooding stress responses. A search for functional molecular markers revealed 4,489 microsatellites and 560,298 single nucleotide polymorphisms (SNPs). A quantitative real-time PCR analysis validated the RNA-seq expression analysis and allowed for the identification of transcriptomic differences between the two evaluated genotypes. Moreover, 192 unannotated sequences were classified as containing complete open reading frames, suggesting that the new, potentially exclusive genes should be further investigated.ConclusionThe present study represents the first whole-transcriptome sequencing of U. humidicola leaves, providing an important public information source of transcripts and functional molecular markers. The qPCR analysis indicated that the expression of certain transcripts confirmed the differential expression observed in silico, which demonstrated that RNA-seq is useful for identifying differentially expressed and unique genes. These results corroborate the findings from previous studies and suggest a hybrid origin for BH031.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-3270-5) contains supplementary material, which is available to authorized users.

Proteins involved in biophoton emission and flooding-stress responses in soybean under light and dark conditions.

To know the molecular systems basically flooding conditions in soybean, biophoton emission measurements and proteomic analyses were carried out for flooding-stressed roots under light and dark conditions. Photon emission was analyzed using a photon counter. Gel-free quantitative proteomics were performed to identify significant changes proteins using the nano LC-MS along with SIEVE software. Biophoton emissions were significantly increased in both light and dark conditions after flooding stress, but gradually decreased with continued flooding exposure compared to the control plants. Among the 120 significantly identified proteins in the roots of soybean plants, 73 and 19 proteins were decreased and increased in the light condition, respectively, and 4 and 24 proteins were increased and decreased, respectively, in the dark condition. The proteins were mainly functionally grouped into cell organization, protein degradation/synthesis, and glycolysis. The highly abundant lactate/malate dehydrogenase proteins were decreased in flooding-stressed roots exposed to light, whereas the lysine ketoglutarate reductase/saccharopine dehydrogenase bifunctional enzyme was increased in both light and dark conditions. Notably, however, specific enzyme assays revealed that the activities of these enzymes and biophoton emission were sharply increased after 3 days of flooding stress. This finding suggests that the source of biophoton emission in roots might involve the chemical excitation of electron or proton through enzymatic or non-enzymatic oxidation and reduction reactions. Moreover, the lysine ketoglutarate reductase/saccharopine dehydrogenase bifunctional enzyme may play important roles in responses in flooding stress of soybean under the light condition and as a contributing factor to biophoton emission.

Jasmonic acid induced protein response to biophoton emissions and flooding stress in soybean

Biophoton emissions were elevated by the exogenous plant hormone application such as jasmonic (JA) and salicylic acid (SA). To reveal the molecular mechanisms underlying flooding stress responses in soybean treated with JA and SA, biophoton emissions from plants were quantified in combination with proteomic analyses. Furthermore, treatment with exogenous JA inhibited lateral root growth and markedly reduced root weight. Out of 649 proteins identified in the JA- and JA/SA-treated plants, 44 were unique to JA-treated plants, 403 were unique to JA/SA-treated plants, and 202 were shared between the groups. These proteins were involved in stress, signaling, degradation, glycolysis, fermentation, and hormone metabolism. The abundances of glutathione-S-transferase, alanine aminotransferase, and malate dehydrogenase were decreased; however, the activities of these enzymes were increased. In contrast, the abundance and activity of monodehydroascorbate reductase increased in the roots of plants treated with JA and SA under flooding stress. This suggests that the quantity of lateral roots, total root mass, and free radicals generated during oxidation and reduction reactions and reactive oxygen species scavenging largely contribute to biophoton emission. Furthermore, monodehydroascorbate reductase, which is involved in detoxification and controlling hydrogen peroxide levels, may protect plant cells against oxidative damage during flooding. Biological significanceTo understand the source of biophoton emission and molecular mechanism by the application of jasmonic and salicylic acid under flooding conditions in soybean plants, the label-free quantitative techniques were performed in roots. Root lengths and weights were significantly reduced by the effect of jasmonic and salicylic acid while it inhibited growth of the lateral roots in normal conditions using the jasmonic acid. Finally, identified proteins were functionally annotated by MAPMAN software application; that were assigned to different functional categories, such as stress, signaling, protein, glycolysis, metabolism, cell wall, and cell organization. Consequently, this study offers to learn the photon emission in plants and to know the molecular mechanism under flooding stress in soybean.

Arabidopsis CML38, a Calcium Sensor That Localizes to Ribonucleoprotein Complexes under Hypoxia Stress.

During waterlogging and the associated oxygen deprivation stress, plants respond by the induction of adaptive programs, including the redirected expression of gene networks toward the synthesis of core hypoxia-response proteins. Among these core response proteins in Arabidopsis (Arabidopsis thaliana) is the calcium sensor CML38, a protein related to regulator of gene silencing calmodulin-like proteins (rgsCaMs). CML38 transcripts are up-regulated more than 300-fold in roots within 6 h of hypoxia treatment. Transfer DNA insertional mutants of CML38 show an enhanced sensitivity to hypoxia stress, with lowered survival and more severe inhibition of root and shoot growth. By using yellow fluorescent protein (YFP) translational fusions, CML38 protein was found to be localized to cytosolic granule structures similar in morphology to hypoxia-induced stress granules. Immunoprecipitation of CML38 from the roots of hypoxia-challenged transgenic plants harboring CML38pro::CML38:YFP followed by liquid chromatography-tandem mass spectrometry analysis revealed the presence of protein targets associated with messenger RNA ribonucleoprotein (mRNP) complexes including stress granules, which are known to accumulate as messenger RNA storage and triage centers during hypoxia. This finding is further supported by the colocalization of CML38 with the mRNP stress granule marker RNA Binding Protein 47 (RBP47) upon cotransfection of Nicotiana benthamiana leaves. Ruthenium Red treatment results in the loss of CML38 signal in cytosolic granules, suggesting that calcium is necessary for stress granule association. These results confirm that CML38 is a core hypoxia response calcium sensor protein and suggest that it serves as a potential calcium signaling target within stress granules and other mRNPs that accumulate during flooding stress responses.

Analysis of Plasma Membrane Proteome in Soybean and Application to Flooding Stress Response

The plasma membrane acts as the primary interface between the cellular cytoplasm and the extracellular environment. To investigate the function of the plasma membrane in response to flooding stress, plasma membrane was purified from root and hypocotyl of soybean seedlings using an aqueous two-phase partitioning method. Purified plasma membrane proteins with 81% purity were analyzed using either two-dimensional polyacrylamide gel electrophoresis followed by mass spectrometry and protein sequencing (2-DE MS/sequencer)-based proteomics or nanoliquid chromatography followed by mass spectrometry (nanoLC-MS/MS)-based proteomics. The number of hydrophobic proteins identified by nanoLC-MS/MS-based proteomics was compared with those identified by 2-DE MS/sequencer-based proteomics. These techniques were applied to identify the proteins in soybean that are responsive to flooding stress. Results indicate insights of plasma membrane into the response of soybean to flooding stress: (i) the proteins located in the cell wall are up-regulated in plasma membrane; (ii) the proteins related to antioxidative system play a crucial role in protecting cells from oxidative damage; (iii) the heat shock cognate protein plays a role in protecting proteins from denaturation and degradation during flooding stress; and (iv) the signaling related proteins might regulate ion homeostasis.

Modelling the impact of flooding stress on the growth performance of woody species using fuzzy logic

Among the driving processes responsible for riparian forest dynamics the species-specific impact of flooding on the development of woody plants plays a key role—particularly for lowland rivers. Only a few of the forest succession models currently in use incorporate the flooding stress response of trees. This situation is mainly due to the incomplete investigation of the flooding tolerance processes and the related abiotic and biotic factors. In an attempt to use the wide-ranging but still rather vague knowledge available on flooding stress, the research presented in this paper proposes an approach to model tree response to flooding using the fuzzy set theory. The model is illustrated for the case of central European species. Flooding stress response to the abiotic factors of duration, depth and frequency of flooding differs according to five flooding tolerance classes and is expressed by means of a growth factor that limits optimal tree growth. We show that existing fuzzy set theory is able to generate and calibrate a flood stress response model which in turn can be incorporated into more complex forest succession models adapted to riparian areas.

Cytosolic ascorbate peroxidase 2 (cAPX 2) is involved in the soybean response to flooding

Proteomic analyses of soybean seedlings responding to flooding were conducted to identify key proteins involved. The seeds were germinated on a spongy matrix for two days, and then subjected to flooding for three days. After flooding, the total number of roots, the length of the main root, the lengths of the lateral and adventitious roots, and the fresh weight of the underground tissues of flooded soybean seedlings were significantly suppressed compared with nontreated plants. To identify the early flooding-responsive proteins, the seedling roots were used for preparing cytosolic and membrane fractions. After two-dimensional polyacrylamide gel electrophoresis and silver staining, 208 proteins were detected, and the levels of 44 were different from those of the control. The expression pattern of 10 proteins among the 44 from six different soybean cultivars confirmed that the 10 were flooding-responsive proteins. One of the 10 proteins was dominantly down-regulated under flooding conditions and was identified as cytosolic ascorbate peroxidase 2 (cAPX 2). Northern-hybridization showed that the abundance of cAPX 2 transcript decreased significantly after flooding, as did the enzymatic activity of APX. These results suggest that cAPX 2 is involved in flooding stress responses in young soybean seedlings.