RESUMO
BACKGROUND: Drought tolerance is a complex quantitative trait that involves the coordination of a vast array of genes belonging to different pathways. To identify genes related to the drought-tolerance pathway in rice, we carried out gene-expression profiling of the leaves of near-isogenic lines (NILs) with similar genetic backgrounds and different set of QTLs but contrasting drought tolerance levels in response to long-term drought-stress treatments. This work will help differentiate mechanisms of tolerance in contrasting NILs and accelerate molecular breeding programs to improve drought tolerance in this crop. RESULTS: The two pairs of rice NILs, developed at the International Rice Research Institute, along with the drought-susceptible parent, IR64, showed distinct gene-expression profiles in leaves under different water-deficit (WD) treatments. Drought tolerance in the highly drought-tolerant NIL (DTN), IR77298-14-1-2-B-10, could be attributed to the up-regulation of genes with calcium ion binding, transferase, hydrolase and transcription factor activities, whereas in the moderate DTN, IR77298-5-6-B-18, genes with transporter, catalytic and structural molecule activities were up-regulated under WD. In IR77298-14-1-2-B-10, the induced genes were characterized by the presence of regulatory motifs in their promoters, including TGGTTAGTACC and ([CT]AAC[GT]G){2}, which are specific to the TFIIIA and Myb transcription factors, respectively. In IR77298-5-6-B-18, promoters containing a GCAC[AG][ACGT][AT]TCCC[AG]A[ACGT]G[CT] motif, common to MADS(AP1), HD-ZIP, AP2 and YABBY, were induced, suggesting that these factors may play key roles in the regulation of drought tolerance in these two DTNs under severe WD. CONCLUSIONS: We report here that the two pairs of NILs with different levels of drought tolerance may elucidate potential mechanisms and pathways through transcriptome data from leaf tissue. The present study serves as a resource for marker discovery and provides detailed insight into the gene-expression profiles of rice leaves, including the main functional categories of drought-responsive genes and the genes that are involved in drought-tolerance mechanisms, to help breeders identify candidate genes (both up- and down-regulated) associated with drought tolerance and suitable targets for manipulating the drought-tolerance trait in rice.
Assuntos
Oryza/genética , Folhas de Planta/genética , Secas , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/genéticaRESUMO
The NAC (NAM, ATAF1/2 and CUC2) genes are plant-specific transcriptional factors known to play diverse roles in various plant developmental processes. We describe the rice (Oryza sativa) OsNAC genes expression profiles (GEPs) under normal and water-deficit treatments (WDTs). The GEPs of the OsNAC genes were analyzed in 25 tissues covering the entire life cycle of Minghui 63. High expression levels of 17 genes were demonstrated in certain tissues under normal conditions suggesting that these genes may play important roles in specific organs. We determined that 16 genes were differentially expressed under at least 1 phytohormone (NAA, GA3, KT, SA, ABA, and JA) treatment. To investigate the GEPs in the root, leaf, and panicle of three rice genotypes [e.g., 2 near-isogenic lines (NILs) and IR64], we used two NILs from a common genetic combination backcross developed by Aday Selection and IR64. WDTs were applied using the fraction of transpirable soil water at severe, mild, and control conditions. Transcriptomic analysis using a 44K oligoarray from Agilent was performed on all the tissue samples. We identified common and specific genes in all tissues from the two NILs under both WDTs, and the majority of the OsNAC genes that were activated were in the drought-tolerant IR77298-14-1-2-B-10 line compared with the drought-susceptible IR77298-14-1-2-B-13 or IR64. In IR77298-14-1-2-B-10, seventeen genes were very specific in their expression levels. Approximately 70 % of the genes from subgroups SNAC and NAM/CUC3 were activated in the leaf, but 37 % genes from subgroup SND were inactivated in the root compared with the control under severe stress conditions. These results provide a useful reference for the cloning of candidate genes from the specific subgroup for further functional analysis.
Assuntos
Genes de Plantas , Oryza/genética , Cruzamentos Genéticos , Secas , Topos Floridos/efeitos dos fármacos , Topos Floridos/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Genes de Plantas/efeitos dos fármacos , Genes Reguladores , Teste de Complementação Genética , Modelos Genéticos , Família Multigênica/efeitos dos fármacos , Oryza/efeitos dos fármacos , Oryza/crescimento & desenvolvimento , Oryza/fisiologia , Reguladores de Crescimento de Plantas/farmacologia , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/genética , Proteínas de Plantas/genética , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/genética , RNA de Plantas/genética , Estresse Fisiológico/genética , Fatores de Transcrição/genéticaRESUMO
The AP2/EREBP genes play various roles in developmental processes and in stress-related responses in plants. Genome-wide microarrays based on the gene expression profiles of the AP2/EREBP family were analyzed under conditions of normal growth and drought stress. The preferential expression of fifteen genes was observed in specific tissues, suggesting that these genes may play important roles in vegetative and reproductive stages of growth. A large number of redundant genes were differentially expressed following phytohormone treatments (NAA, GA3, KT, SA, JA, and ABA). To investigate the gene expression responses in the root, leaf, and panicle of three rice genotypes, two drought stress conditions were applied using the fraction of transpirable soil water (FTSW) under severe (0.2 FTSW), mild (0.5 FTSW), and control (1.0 FTSW) conditions. Following treatment, transcriptomic analysis using a 44-K oligoarray from Agilent was performed on all the tissue samples. We identified common and specific genes in all tissues from two near-isogenic lines, IR77298-14-1-2-B-10 (drought tolerant) and IR77298-14-1-2-B-13 (drought susceptible), under drought stress conditions. The majority of the genes that were activated in the IR77298-14-1-2-B-10 line were members of the AP2/EREBP gene family. Non-redundant genes (sixteen) were found in the drought-tolerant line, and four genes were selected as candidate novel reference genes because of their higher expression levels in IR77298-14-1-2-B-10. Most of the genes in the AP2, B3, and B5 subgroups were involved in the panicle under severe stress conditions, but genes from the B1 and B2 subgroups were down-regulated in the root. Of the four subfamilies, RAV exhibited the highest number of up-regulated genes (80%) in the panicle under severe stress conditions in the drought-tolerant line compared to Minghui 63 under normal conditions, and the gene structures of the RAV subfamily may be involved in the response to drought stress in the flowering stage. These results provide a useful reference for the cloning of candidate genes from the specific subgroup for further functional analysis.
Assuntos
Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Secas , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Família Multigênica/genética , Oryza/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Estresse Fisiológico/fisiologia , Flores/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Genótipo , Análise em Microsséries , Oryza/crescimento & desenvolvimento , Oryza/metabolismo , Reguladores de Crescimento de Plantas/farmacologia , Folhas de Planta/metabolismo , Raízes de Plantas/metabolismo , Locos de Características Quantitativas/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Especificidade da Espécie , Estresse Fisiológico/genéticaRESUMO
Lowland rice roots have a unique physiological response to drought because of their adaptation to flooded soil. Rice root attributes that facilitate growth under flooded conditions may affect rice response to drought, but the relative roles of root structural and functional characteristics for water uptake under drought in rice are not known. Morphological, anatomical, biochemical, and molecular attributes of soil-grown rice roots were measured to investigate the genotypic variability and genotype×environment interactions of water uptake under variable soil water regimes. Drought-resistant genotypes had the lowest night-time bleeding rates of sap from the root system in the field. Diurnal fluctuation predominated as the strongest source of variation for bleeding rates in the field and root hydraulic conductivity (Lpr) in the greenhouse, and was related to expression trends of various PIP and TIP aquaporins. Root anatomy was generally more responsive to drought treatments in drought-resistant genotypes. Suberization and compaction of sclerenchyma layer cells decreased under drought, whereas suberization of the endodermis increased, suggesting differential roles of these two cell layers for the retention of oxygen under flooded conditions (sclerenchyma layer) and retention of water under drought (endodermis). The results of this study point to the genetic variability in responsiveness to drought of rice roots in terms of morphology, anatomy, and function.
Assuntos
Adaptação Fisiológica/fisiologia , Oryza/fisiologia , Raízes de Plantas/fisiologia , Transpiração Vegetal/fisiologia , Água/metabolismo , Aquaporinas/genética , Transporte Biológico , Análise por Conglomerados , Secas , Genótipo , Oryza/anatomia & histologia , Oryza/crescimento & desenvolvimento , Oryza/metabolismo , Proteínas de Plantas/genética , Raízes de Plantas/anatomia & histologia , Raízes de Plantas/metabolismo , Brotos de Planta/anatomia & histologia , Brotos de Planta/crescimento & desenvolvimento , Brotos de Planta/metabolismo , Brotos de Planta/fisiologia , Solo/químicaRESUMO
BACKGROUND: Plant roots are important organs to uptake soil water and nutrients, perceiving and transducing of soil water deficit signals to shoot. The current knowledge of drought stress transcriptomes in rice are mostly relying on comparative studies of diverse genetic background under drought. A more reliable approach is to use near-isogenic lines (NILs) with a common genetic background but contrasting levels of resistance to drought stress under initial exposure to water deficit. Here, we examined two pairs of NILs in IR64 background with contrasting drought tolerance. We obtained gene expression profile in roots of rice NILs under different levels of drought stress help to identify genes and mechanisms involved in drought stress. RESULTS: Global gene expression analysis showed that about 55% of genes differentially expressed in roots of rice in response to drought stress treatments. The number of differentially expressed genes (DEGs) increased in NILs as the level of water deficits, increased from mild to severe condition, suggesting that more genes were affected by increasing drought stress. Gene onthology (GO) test and biological pathway analysis indicated that activated genes in the drought tolerant NILs IR77298-14-1-2-B-10 and IR77298-5-6-B-18 were mostly involved in secondary metabolism, amino acid metabolism, response to stimulus, defence response, transcription and signal transduction, and down-regulated genes were involved in photosynthesis and cell wall growth. We also observed gibberellic acid (GA) and auxin crosstalk modulating lateral root formation in the tolerant NILs. CONCLUSIONS: Transcriptome analysis on two pairs of NILs with a common genetic background (~97%) showed distinctive differences in gene expression profiles and could be effective to unravel genes involved in drought tolerance. In comparison with the moderately tolerant NIL IR77298-5-6-B-18 and other susceptible NILs, the tolerant NIL IR77298-14-1-2-B-10 showed a greater number of DEGs for cell growth, hormone biosynthesis, cellular transports, amino acid metabolism, signalling, transcription factors and carbohydrate metabolism in response to drought stress treatments. Thus, different mechanisms are achieving tolerance in the two tolerant lines.
Assuntos
Secas , Oryza/genética , Raízes de Plantas/genética , Transcriptoma , Regulação da Expressão Gênica de Plantas , Genótipo , Análise em Microsséries , Oryza/metabolismo , Raízes de Plantas/metabolismo , RNA de Plantas/genética , Estresse FisiológicoRESUMO
Rice is known to be sensitive to soil water deficit and evaporative demand, with a greatest sensitivity of lowland-adapted genotypes. We have analysed the responses of plant water relations and of leaf elongation rate (LER) to soil water status and evaporative demand in seven rice genotypes belonging to different species, subspecies, either upland- or lowland-adapted. In the considered range of soil water potential (0 to -0.6 MPa), stomatal conductance was controlled in such a way that the daytime leaf water potential was similar in well-watered, droughted or flooded conditions (isohydric behaviour). A low sensitivity of LER to evaporative demand was observed in the same three conditions, with small differences between genotypes and lower sensitivity than in maize. The sensitivity of LER to soil water deficit was similar to that of maize. A tendency towards lower sensitivities was observed in upland than lowland genotypes but with smaller differences than expected. We conclude that leaf water status and leaf elongation of rice are not particularly sensitive to water deficit. The main origin of drought sensitivity in rice may be its poor root system, whose effect was alleviated in the study presented here by growing plants in pots whose soil was entirely colonized by roots of all genotypes.
Assuntos
Oryza/fisiologia , Folhas de Planta/crescimento & desenvolvimento , Transpiração Vegetal , Água/fisiologia , Adaptação Fisiológica , Secas , Genótipo , Oryza/genética , Oryza/crescimento & desenvolvimento , Folhas de Planta/fisiologia , Raízes de Plantas/crescimento & desenvolvimento , Solo/análise , Zea mays/genética , Zea mays/crescimento & desenvolvimento , Zea mays/fisiologiaRESUMO
Genotypic variations in leaf gas exchange and yield were analysed in five upland-adapted and three lowland rice cultivars subjected to a differential soil moisture gradient, varying from well-watered to severely water-stressed conditions. A reduction in the amount of water applied resulted in a significant decrease in leaf gas exchange and, subsequently, in above-ground dry mass and grain yield, that varied among genotypes and distance from the line source. The comparison between the variable J and the Delta values in recently synthesized sugars methods, yielded congruent estimations of mesophyll conductance (g(m)), confirming the reliability of these two techniques. Our data demonstrate that g(m) is a major determinant of photosynthesis (A), because rice genotypes with inherently higher g(m) were capable of keeping higher A in stressed conditions. Furthermore, A, g(s), and g(m) of water-stressed genotypes rapidly recovered to the well-watered values upon the relief of water stress, indicating that drought did not cause any lasting metabolic limitation to photosynthesis. The comparisons between the A/C(i) and corresponding A/C(c) curves, measured in the genotypes that showed intrinsically higher and lower instantaneous A, confirmed this finding. Moreover, the effect of drought stress on grain yield was correlated with the effects on both A and total diffusional limitations to photosynthesis. Overall, these data indicate that genotypes which showed higher photosynthesis and conductances were also generally more productive across the entire soil moisture gradient. The analysis of Delta revealed a substantial variation of water use efficiency among the genotypes, both on the long-term (leaf pellet analysis) and short-term scale (leaf soluble sugars analysis).
Assuntos
Dióxido de Carbono/metabolismo , Isótopos de Carbono/metabolismo , Oryza/crescimento & desenvolvimento , Oryza/genética , Folhas de Planta/metabolismo , Água/metabolismo , Fenômenos Biofísicos , Genótipo , Oryza/química , Oryza/metabolismo , Fotossíntese , Folhas de Planta/química , ReproduçãoRESUMO
The rapid progress in rice genotyping must be matched by advances in phenotyping. A better understanding of genetic variation in rice for drought response, root traits, and practical methods for studying them are needed. In this study, the OryzaSNP set (20 diverse genotypes that have been genotyped for SNP markers) was phenotyped in a range of field and container studies to study the diversity of rice root growth and response to drought. Of the root traits measured across more than 20 root experiments, root dry weight showed the most stable genotypic performance across studies. The environment (E) component had the strongest effect on yield and root traits. We identified genomic regions correlated with root dry weight, percent deep roots, maximum root depth, and grain yield based on a correlation analysis with the phenotypes and aus, indica, or japonica introgression regions using the SNP data. Two genomic regions were identified as hot spots in which root traits and grain yield were co-located; on chromosome 1 (39.7-40.7 Mb) and on chromosome 8 (20.3-21.9 Mb). Across experiments, the soil type/ growth medium showed more correlations with plant growth than the container dimensions. Although the correlations among studies and genetic co-location of root traits from a range of study systems points to their potential utility to represent responses in field studies, the best correlations were observed when the two setups had some similar properties. Due to the co-location of the identified genomic regions (from introgression block analysis) with QTL for a number of previously reported root and drought traits, these regions are good candidates for detailed characterization to contribute to understanding rice improvement for response to drought. This study also highlights the utility of characterizing a small set of 20 genotypes for root growth, drought response, and related genomic regions.
Assuntos
Oryza/crescimento & desenvolvimento , Oryza/genética , Mapeamento Cromossômico , Cromossomos de Plantas/genética , Secas , Meio Ambiente , Estudos de Associação Genética , Genoma de Planta , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Polimorfismo de Nucleotídeo Único , Locos de Características QuantitativasRESUMO
Nitrogen fixation and nodule permeability to O2 diffusion are decreased by drought stress. Since γ-aminobutyric acid (GABA) synthesis is rapidly stimulated by a variety of stress conditions including hypoxia, it was hypothesized that decreased O2 availability in nodules stimulates glutamate decarboxylase (GAD) activity (EC 4.1.1.15), thereby resulting in GABA accumulation. First, the amino acid composition of xylem sap was determined in plants subjected to soil water deficits. While the xylem sap concentration of several amino acids increased when the plant was subjected to a water deficit, the greatest increase was in GABA. GABA accumulation was examined in response to stress induced by hypoxia or the addition of polyethylene glycol (PEG) to the nutrient solution. The exposure of soybean nodules to hypoxia for 6 h enhanced the GABA concentration by 6-fold, but there was no change in GABA concentration in response to the PEG treatment. No major changes in the in vitro GAD activity were measured in nodule cytosol or bacteroids. The present data do not support the hypothesis that decreased nodule O2 permeability and a resulting O2 deprivation inside nodules may stimulate in vitro GAD activity and thus GABA accumulation. However, the data could indicate a possible effect of hypoxia and drought stress on the in vivo activity of GAD.
RESUMO
Drought stress is one of the major factors affecting nitrogen fixation by legume-rhizobium symbiosis. Several mechanisms have been previously reported to be involved in the physiological response of symbiotic nitrogen fixation to drought stress, i.e. carbon shortage and nodule carbon metabolism, oxygen limitation, and feedback regulation by the accumulation of N fixation products. The carbon shortage hypothesis was previously investigated by studying the combined effects of CO2 enrichment and water deficits on nodulation and N2 fixation in soybean. Under drought, in a genotype with drought tolerant N2 fixation, approximately four times the amount of 14C was allocated to nodules compared to a drought sensitive genotype. It was found that an important effect of CO2 enrichment of soybean under drought was an enhancement of photo assimilation, an increased partitioning of carbon to nodules, whose main effect was to sustain nodule growth, which helped sustain N2 rates under soil water deficits. The interaction of nodule permeability to O2 and drought stress with N2 fixation was examined in soybean nodules and led to the overall conclusion that O2 limitation seems to be involved only in the initial stages of water deficit stresses in decreasing nodule activity. The involvement of ureides in the drought response of N2 fixation was initially suspected by an increased ureide concentration in shoots and nodules under drought leading to a negative feedback response between ureides and nodule activity. Direct evidence for inhibition of nitrogenase activity by its products, ureides and amides, supported this hypothesis. The overall conclusion was that all three physiological mechanisms are important in understanding the regulation of N2 fixation and its response of to soil drying.
Assuntos
Desastres , Fabaceae/microbiologia , Fixação de Nitrogênio/fisiologia , Rhizobium/fisiologia , Simbiose/fisiologia , Dióxido de Carbono/análise , SoloRESUMO
Global production of rice (Oryza sativa) grain is limited by water availability and the low 'leaf-level' photosynthetic capacity of many cultivars. Oryza sativa is extremely susceptible to water-deficits; therefore, predicted increases in the frequency and duration of drought events, combined with future rises in global temperatures and food demand, necessitate the development of more productive and drought tolerant cultivars. We investigated the underlying physiological, isotopic and morphological responses to water-deficit in seven common varieties of O. sativa, subjected to prolonged drought of varying intensities, for phenotyping purposes in open field conditions. Significant variation was observed in leaf-level photosynthesis rates (A) under both water treatments. Yield and A were influenced by the conductance of the mesophyll layer to CO2 (g(m)) and not by stomatal conductance (g(s)). Mesophyll conductance declined during drought to differing extents among the cultivars; those varieties that maintained g(m) during water-deficit sustained A and yield to a greater extent. However, the variety with the highest g(m) and yield under well-watered conditions (IR55419-04) was distinct from the most effective cultivar under drought (Vandana). Mesophyll conductance most effectively characterises the photosynthetic capacity and yield of O. sativa cultivars under both well-watered and water-deficit conditions; however, the desired attributes of high g(m) during optimal growth conditions and the capacity for g(m) to remain constant during water-deficit may be mutually exclusive. Nonetheless, future genetic and physiological studies aimed at enhancing O. sativa yield and drought stress tolerance should investigate the biochemistry and morphology of the interface between the sub-stomatal pore and mesophyll layer.
Assuntos
Secas , Flores/fisiologia , Oryza/fisiologia , Fotossíntese , Estresse Fisiológico , Dióxido de Carbono/metabolismo , Isótopos de Carbono , Difusão , Células do Mesofilo/metabolismo , Estômatos de Plantas/fisiologia , Transpiração Vegetal , ÁguaRESUMO
Inhibition of leaf elongation and expansion is one of the earliest responses of rice to water deficit. Despite this sensitivity, a great deal of genetic variation exists in the extant of leaf elongation rate (LER) reduction in response to declining soil moisture. We analyzed global gene expression in the leaf elongation zone under drought in two rice cultivars with disparate LER sensitivities to water stress. We found little overlap in gene regulation between the two varieties under moderate drought; however, the transcriptional response to severe drought was more conserved. In response to moderate drought, we found several genes related to secondary cell wall deposition that were down regulated in Moroberekan, an LER tolerant variety, but up-regulated in LER sensitive variety IR64.
Assuntos
Parede Celular/genética , Secas , Regulação da Expressão Gênica de Plantas , Oryza/genética , Folhas de Planta/genética , Proteínas de Plantas/genética , Fatores de Transcrição/genética , Perfilação da Expressão Gênica , Oryza/anatomia & histologia , Oryza/citologia , Folhas de Planta/anatomia & histologia , Folhas de Planta/citologia , Proteínas de Plantas/metabolismo , Estresse Fisiológico/genética , Fatores de Transcrição/metabolismo , Transcrição GênicaRESUMO
In addition to characterising root architecture, evaluating root water uptake ability is important for understanding drought response. A series of three lysimeter studies were conducted using the OryzaSNP panel, which consists of 20 diverse rice (Oryza sativa L.) genotypes. Large genotypic differences in drought response were observed in this genotype panel in terms of plant growth and water uptake. Total water uptake and daily water uptake rates in the drought-stress treatment were correlated with root length density, especially at depths below 30cm. Patterns of water uptake among genotypes remained consistent throughout the stress treatments: genotypes that initially extracted more water were the same genotypes that extracted more water at the end of the study. These results suggest that response to drought by deep root growth, rather than a conservative soil water pattern, seems to be important for lowland rice. Genotypes in the O. sativa type aus group showed some of the greatest water uptake and root growth values. Since the OryzaSNP panel has been genotyped in detail with SNP markers, we expect that these results will be useful for understanding the genetics of rice root growth and function for water uptake in response to drought.
RESUMO
Drought is a major constraint to rainfed rice production in some parts of sub Saharan Africa (SSA) because of irregular and low rainfall. Improving root water uptake during progressive soil drying could contribute to terminal drought stress resistance. Two lysimeter experiments were conducted using 200 introgression lines (ILs) derived from Oryza sativa L.×Oryza glaberrima Steud. crosses (IR64/RAM54 and IR64/RAM90) to characterise water uptake and root traits under progressive soil drying and to assess genetic differences among these lines in relation to drought stress. The lysimeter facility setup and protocols for simultaneous monitoring of water uptake rates and leaf area are described. Significant genetic variability for water uptake and root traits among the ILs was observed under drought stress. Water uptake in the drought stress treatments was correlated with root length density (RLD) and root dry matter (RDM) at lower soil depths. The highest RLD values at depth were observed among the RAM90 ILs and genetic differences were observed between RAM90 and RAM54 populations, implying that accession RAM90 could be used as a source for improving drought avoidance in lowland rice. The ILs with greater water uptake identified in this study will be used in future research for improving drought resistance of rice in SSA.
RESUMO
Thermal imaging using infrared (IR) is now an established technology for the study of stomatal responses and for phenotyping plants for differences in stomatal behaviour. This paper outlines the potential applications of IR sensing in drought phenotyping, with particular emphasis on a description of the problems with extrapolation of the technique from the study of single leaves in controlled environments to the study of plant canopies is field plots, with examples taken from studies on grapevine (Vitis vinifera L.) and rice (Oryza sativa L.). Particular problems include the sensitivity of leaf temperature (and potentially the temperature of reference surfaces) to both temporal and spatial variation in absorbed radiation, with leaf temperature varying by as much as 15°C between full sun and deep shade. Examples of application of the approach to phenotyping in the field and the steps in data analysis are outlined, demonstrating that clear genotypic variation may be detected despite substantial variation in soil moisture status or incident radiation by the use of appropriate normalisation techniques.
RESUMO
UNLABELLED: BACKGROUND AND AIMS The inner cortical cells (IC-cells) of legume root nodules have been previously shown to regulate the resistance to nodule O2 diffusion by a rapid contraction/expansion mechanism, which controls the volume of intercellular spaces and their occlusion by a liquid phase. The expression of aquaporins in IC-cells was also found to be involved in this nodule O2 diffusion mechanism. The aim of this study was to compare the expression of plasma membrane intrinsic proteins (PIP) aquaporin isoforms with tonoplast intrinsic protein (gamma-TIP) in both IC-cells and adjacent cell types. METHODS: Using immunogold labelling in ultra-thin sections of Glycine max nodules, the expression of two PIP isoforms was observed and compared with the gamma-TIP pattern. KEY RESULTS: The plasma membrane aquaporins PIP1 and PIP2 were expressed more in IC-cells and endodermis than in pericycle and infected cells. The tonoplast aquaporin gamma-TIP has shown a distribution pattern similar to that of the PIPs. CONCLUSIONS: PIPs and gamma-TIP aquaporins are highly expressed in both plasmalemma and tonoplast of nodule IC-cells. This distribution is consistent with the putative role of water fluxes associated with the regulation of nodule conductance to O2 diffusion and the subsequent ATP-dependent nitrogenase activity. In the endodermis, these aquaporins might also be involved in nutrient transport between the infected zone and vascular traces.
Assuntos
Aquaporinas/metabolismo , Glycine max/metabolismo , Proteínas de Plantas/metabolismo , Raízes de Plantas/microbiologia , Regulação da Expressão Gênica de Plantas/fisiologia , Raízes de Plantas/metabolismo , Glycine max/microbiologia , Distribuição Tecidual , Água/metabolismoRESUMO
The hypothesis that elevated [CO(2)] alleviates ureide inhibition of N(2)-fixation was tested. Short-term responses of the acetylene reduction assay (ARA), ureide accumulation and total non-structural carbohydrate (TNC) levels were measured following addition of ureide to the nutrient solution of hydroponically grown soybean. The plants were exposed to ambient (360 micromol mol(-1)) or elevated (700 micromol mol(-1)) [CO(2)]. Addition of 5 and 10 mM ureide to the nutrient solution inhibited N(2)-fixation activity under both ambient and elevated [CO(2)] conditions. However, the percentage inhibition following ureide treatment was significantly greater under ambient [CO(2)] as compared with that under elevated [CO(2)]. Under ambient [CO(2)] conditions, ARA was less than that under elevated [CO(2)] 1 d after ureide treatment. Under ambient [CO(2)], the application of ureide resulted in a significant accumulation of ureide in all plant tissues, with the highest concentration increases in the leaves. However, application of exogenous ureide to plants subjected to elevated [CO(2)] did not result in increased ureide concentration in any tissues. TNC concentrations were consistently higher under elevated [CO(2)] compared with those under ambient [CO(2)]. For both [CO(2)] treatments, the application of ureide induced a significant decrease of TNC concentrations in the leaves and nodules. For both leaves and nodules, a negative correlation was observed between TNC and ureide levels. Results indicate that product(s) of ureide catabolism rather than tissue ureide concentration itself are critical in the regulation of N(2)-fixation.