RESUMEN
Microorganisms produce volatile compounds (VCs) that promote plant growth and photosynthesis through complex mechanisms involving cytokinin (CK) and abscisic acid (ABA). We hypothesized that plants' responses to microbial VCs involve posttranslational modifications of the thiol redox proteome through action of plastidial NADPH-dependent thioredoxin reductase C (NTRC), which regulates chloroplast redox status via its functional relationship with 2-Cys peroxiredoxins. To test this hypothesis, we analysed developmental, metabolic, hormonal, genetic, and redox proteomic responses of wild-type (WT) plants and a NTRC knockout mutant (ntrc) to VCs emitted by the phytopathogen Alternaria alternata. Fungal VC-promoted growth, changes in root architecture, shifts in expression of VC-responsive CK- and ABA-regulated genes, and increases in photosynthetic capacity were substantially weaker in ntrc plants than in WT plants. As in WT plants, fungal VCs strongly promoted growth, chlorophyll accumulation, and photosynthesis in ntrc-Δ2cp plants with reduced 2-Cys peroxiredoxin expression. OxiTRAQ-based quantitative and site-specific redox proteomic analyses revealed that VCs promote global reduction of the thiol redox proteome (especially of photosynthesis-related proteins) of WT leaves but its oxidation in ntrc leaves. Our findings show that NTRC is an important mediator of plant responses to microbial VCs through mechanisms involving global thiol redox proteome changes that affect photosynthesis.
Asunto(s)
Alternaria , Proteínas de Arabidopsis/metabolismo , Arabidopsis/efectos de los fármacos , Procesamiento Proteico-Postraduccional/efectos de los fármacos , Reductasa de Tiorredoxina-Disulfuro/metabolismo , Compuestos Orgánicos Volátiles/farmacología , Ácido Abscísico/metabolismo , Arabidopsis/metabolismo , Citocininas/metabolismo , Fotosíntesis/efectos de los fármacos , ProteomaRESUMEN
Volatile compounds (VCs) emitted by phylogenetically diverse microorganisms (including plant pathogens and microbes that do not normally interact mutualistically with plants) promote photosynthesis, growth, and the accumulation of high levels of starch in leaves through cytokinin (CK)-regulated processes. In Arabidopsis (Arabidopsis thaliana) plants not exposed to VCs, plastidic phosphoglucose isomerase (pPGI) acts as an important determinant of photosynthesis and growth, likely as a consequence of its involvement in the synthesis of plastidic CKs in roots. Moreover, this enzyme plays an important role in connecting the Calvin-Benson cycle with the starch biosynthetic pathway in leaves. To elucidate the mechanisms involved in the responses of plants to microbial VCs and to investigate the extent of pPGI involvement, we characterized pPGI-null pgi1-2 Arabidopsis plants cultured in the presence or absence of VCs emitted by Alternaria alternata We found that volatile emissions from this fungal phytopathogen promote growth, photosynthesis, and the accumulation of plastidic CKs in pgi1-2 leaves. Notably, the mesophyll cells of pgi1-2 leaves accumulated exceptionally high levels of starch following VC exposure. Proteomic analyses revealed that VCs promote global changes in the expression of proteins involved in photosynthesis, starch metabolism, and growth that can account for the observed responses in pgi1-2 plants. The overall data show that Arabidopsis plants can respond to VCs emitted by phytopathogenic microorganisms by triggering pPGI-independent mechanisms.
Asunto(s)
Alternaria/química , Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimología , Arabidopsis/microbiología , Glucosa-6-Fosfato Isomerasa/metabolismo , Plastidios/enzimología , Compuestos Orgánicos Volátiles/farmacología , Alternaria/efectos de la radiación , Arabidopsis/crecimiento & desarrollo , Arabidopsis/fisiología , Pared Celular/metabolismo , Pared Celular/efectos de la radiación , Citocininas/metabolismo , Luz , Células del Mesófilo/efectos de los fármacos , Células del Mesófilo/metabolismo , Células del Mesófilo/efectos de la radiación , Mutación/genética , Fotosíntesis/efectos de la radiación , Plastidios/efectos de los fármacos , Proteoma/metabolismo , Almidón/metabolismoRESUMEN
It is known that volatile emissions from some beneficial rhizosphere microorganisms promote plant growth. Here we show that volatile compounds (VCs) emitted by phylogenetically diverse rhizosphere and non-rhizhosphere bacteria and fungi (including plant pathogens and microbes that do not normally interact mutualistically with plants) promote growth and flowering of various plant species, including crops. In Arabidopsis plants exposed to VCs emitted by the phytopathogen Alternaria alternata, changes included enhancement of photosynthesis and accumulation of high levels of cytokinins (CKs) and sugars. Evidence obtained using transgenic Arabidopsis plants with altered CK status show that CKs play essential roles in this phenomenon, because growth and flowering responses to the VCs were reduced in mutants with CK-deficiency (35S:AtCKX1) or low receptor sensitivity (ahk2/3). Further, we demonstrate that the plant responses to fungal VCs are light-dependent. Transcriptomic analyses of Arabidopsis leaves exposed to A. alternata VCs revealed changes in the expression of light- and CK-responsive genes involved in photosynthesis, growth and flowering. Notably, many genes differentially expressed in plants treated with fungal VCs were also differentially expressed in plants exposed to VCs emitted by the plant growth promoting rhizobacterium Bacillus subtilis GB03, suggesting that plants react to microbial VCs through highly conserved regulatory mechanisms.
Asunto(s)
Citocininas/fisiología , Flores/crecimiento & desarrollo , Desarrollo de la Planta/fisiología , Plantas/microbiología , Compuestos Orgánicos Volátiles/metabolismo , Alternaria/fisiología , Arabidopsis/microbiología , Arabidopsis/fisiología , Flores/fisiología , Regulación de la Expresión Génica de las Plantas/fisiología , Fotosíntesis/fisiología , Reacción en Cadena en Tiempo Real de la Polimerasa , Rizosfera , Transcriptoma/fisiologíaRESUMEN
Repeated measurements of crop height to observe plant growth dynamics in real field conditions represent a challenging task. Although there are ways to collect data using sensors on UAV systems, proper data processing and analysis are the key to reliable results. As there is need for specialized software solutions for agricultural research and breeding purposes, we present here a fast algorithm ALFA for the processing of UAV LiDAR derived point-clouds to extract the information on crop height at many individual cereal field-plots at multiple time points. Seven scanning flights were performed over 3 blocks of experimental barley field plots between April and June 2021. Resulting point-clouds were processed by the new algorithm ALFA. The software converts point-cloud data into a digital image and extracts the traits of interest-the median crop height at individual field plots. The entire analysis of 144 field plots of dimension 80 x 33 meters measured at 7 time points (approx. 100 million LiDAR points) takes about 3 minutes at a standard PC. The Root Mean Square Deviation of the software-computed crop height from the manual measurement is 5.7 cm. Logistic growth model is fitted to the measured data by means of nonlinear regression. Three different ways of crop-height data visualization are provided by the software to enable further analysis of the variability in growth parameters. We show that the presented software solution is a fast and reliable tool for automatic extraction of plant height from LiDAR images of individual field-plots. We offer this tool freely to the scientific community for non-commercial use.
Asunto(s)
Hordeum , Fitomejoramiento , Programas Informáticos , Algoritmos , Agricultura/métodosRESUMEN
BACKGROUND: Plants, like all living organisms, metamorphose their bodies during their lifetime. All the developmental and growth events in a plant's life are connected to specific points in time, be it seed germination, seedling emergence, the appearance of the first leaf, heading, flowering, fruit ripening, wilting, or death. The onset of automated phenotyping methods has brought an explosion of such time-to-event data. Unfortunately, it has not been matched by an explosion of adequate data analysis methods. RESULTS AND DISCUSSION: In this paper, we introduce the Bayesian approach towards time-to-event data in plant biology. As a model example, we use seedling emergence data of maize under control and stress conditions but the Bayesian approach is suitable for any time-to-event data (see the examples above). In the proposed framework, we are able to answer key questions regarding plant emergence such as these: (1) Do seedlings treated with compound A emerge earlier than the control seedlings? (2) What is the probability of compound A increasing seedling emergence by at least 5 percent? CONCLUSION: Proper data analysis is a fundamental task of general interest in life sciences. Here, we present a novel method for the analysis of time-to-event data which is applicable to many plant developmental parameters measured in field or in laboratory conditions. In contrast to recent and classical approaches, our Bayesian computational method properly handles uncertainty in time-to-event data and it is capable to reliably answer questions that are difficult to address by classical methods.
RESUMEN
Plant phenotyping platforms offer automated, fast scoring of traits that simplify the selection of varieties that are more competitive under stress conditions. However, indoor phenotyping methods are frequently based on the analysis of plant growth in individual pots. We present a reproducible indoor phenotyping method for screening young barley populations under water stress conditions and after subsequent rewatering. The method is based on a simple read-out of data using RGB imaging, projected canopy height, as a useful feature for indirectly following the kinetics of growth and water loss in a population of barley. A total of 47 variables including 15 traits and 32 biochemical metabolites measured (morphometric parameters, chlorophyll fluorescence imaging, quantification of stress-related metabolites; amino acids and polyamines, and enzymatic activities) were used to validate the method. The study allowed the identification of metabolites related to water stress response and recovery. Specifically, we found that cadaverine (Cad), 1,3-aminopropane (DAP), tryptamine (Tryp), and tyramine (Tyra) were the major contributors to the water stress response, whereas Cad, DAP, and Tyra, but not Tryp, remained at higher levels in the stressed plants even after rewatering. In this work, we designed, optimized and validated a non-invasive image-based method for automated screening of potential water stress tolerance genotypes in barley populations. We demonstrated the applicability of the method using transgenic barley lines with different sensitivity to drought stress showing that combining canopy height and the metabolite profile we can discriminate tolerant from sensitive genotypes. We showed that the projected canopy height a sensitive trait that truly reflects other invasively studied morphological, physiological, and metabolic traits and that our presented methodological setup can be easily applicable for large-scale screenings in low-cost systems equipped with a simple RGB camera. We believe that our approach will contribute to accelerate the study and understanding of the plant water stress response and recovery capacity in crops, such as barley.
RESUMEN
Environmental stresses have a significant effect on agricultural crop productivity worldwide. Exposure of seeds to abiotic stresses, such as salinity among others, results in lower seed viability, reduced germination, and poor seedling establishment. Alternative agronomic practices, e.g., the use of plant biostimulants, have attracted considerable interest from the scientific community and commercial enterprises. Biostimulants, i.e., products of biological origin (including bacteria, fungi, seaweeds, higher plants, or animals) have significant potential for (i) improving physiological processes in plants and (ii) stimulating germination, growth and stress tolerance. However, biostimulants are diverse, and can range from single compounds to complex matrices with different groups of bioactive components that have only been partly characterized. Due to the complex mixtures of biologically active compounds present in biostimulants, efficient methods for characterizing their potential mode of action are needed. In this study, we report the development of a novel complex approach to biological activity testing, based on multi-trait high-throughput screening (MTHTS) of Arabidopsis characteristics. These include the in vitro germination rate, early seedling establishment capacity, growth capacity under stress and stress response. The method is suitable for identifying new biostimulants and characterizing their mode of action. Representatives of compatible solutes such as amino acids and polyamines known to be present in many of the biostimulant irrespective of their origin, i.e., well-established biostimulants that enhance stress tolerance and crop productivity, were used for the assay optimization and validation. The selected compounds were applied through seed priming over a broad concentration range and the effect was investigated simultaneously under control, moderate stress and severe salt stress conditions. The new MTHTS approach represents a powerful tool in the field of biostimulant research and development and offers direct classification of the biostimulants mode of action into three categories: (1) plant growth promotors/inhibitors, (2) stress alleviators, and (3) combined action.
RESUMEN
Physiologically, abscisic acid (ABA) is believed to be a general inhibitor of plant growth, including during the crucial early development of seedlings. However, this view contradicts many reports of stimulatory effects of ABA that, so far, have not been considered in the debate concerning ABA's function in plant development. To address this apparent contradiction, we propose a hypothetical mechanism to explain how ABA might contribute to the promotion of cell expansion. We wish to overturn conventional views on ABA's role during juvenile plant development and put forward the idea that, as for other phytohormones, the role of ABA is determined by dose and sensitivity and ranges from stimulatory to inhibitory effects.
Asunto(s)
Ácido Abscísico/metabolismo , Desarrollo de la Planta , Reguladores del Crecimiento de las Plantas/metabolismo , Plantones/fisiología , Plantones/crecimiento & desarrolloRESUMEN
High-throughput plant phenotyping platforms provide new possibilities for automated, fast scoring of several plant growth and development traits, followed over time using non-invasive sensors. Using Arabidopsis as a model offers important advantages for high-throughput screening with the opportunity to extrapolate the results obtained to other crops of commercial interest. In this study we describe the development of a highly reproducible high-throughput Arabidopsis in vitro bioassay established using our OloPhen platform, suitable for analysis of rosette growth in multi-well plates. This method was successfully validated on example of multivariate analysis of Arabidopsis rosette growth in different salt concentrations and the interaction with varying nutritional composition of the growth medium. Several traits such as changes in the rosette area, relative growth rate, survival rate and homogeneity of the population are scored using fully automated RGB imaging and subsequent image analysis. The assay can be used for fast screening of the biological activity of chemical libraries, phenotypes of transgenic or recombinant inbred lines, or to search for potential quantitative trait loci. It is especially valuable for selecting genotypes or growth conditions that improve plant stress tolerance.
RESUMEN
Current methods of in-house plant phenotyping are providing a powerful new tool for plant biology studies. The self-constructed and commercial platforms established in the last few years, employ non-destructive methods and measurements on a large and high-throughput scale. The platforms offer to certain extent, automated measurements, using either simple single sensor analysis, or advanced integrative simultaneous analysis by multiple sensors. However, due to the complexity of the approaches used, it is not always clear what such forms of plant phenotyping can offer the potential end-user, i.e. plant biologist. This review focuses on imaging methods used in the phenotyping of plant shoots including a brief survey of the sensors used. To open up this topic to a broader audience, we provide here a simple introduction to the principles of automated non-destructive analysis, namely RGB, chlorophyll fluorescence, thermal and hyperspectral imaging. We further on present an overview on how and to which extent, the automated integrative in-house phenotyping platforms have been used recently to study the responses of plants to various changing environments.
RESUMEN
The role of abscisic acid (ABA) during early development was investigated in tomato seedlings. The endogenous content of ABA in particular organs was analyzed in seedlings grown in the dark and under blue light. Our results showed that in dark-grown seedlings, the ABA accumulation was maximal in the cotyledons and elongation zone of hypocotyl, whereas under blue-light, the ABA content was distinctly reduced. Our data are consistent with the conclusion that ABA promotes the growth of etiolated seedlings and the results suggest that ABA plays an inhibitory role in de-etiolation and photomorphogenesis in tomato.
Asunto(s)
Ácido Abscísico/metabolismo , Cotiledón/crecimiento & desarrollo , Etiolado , Hipocótilo/crecimiento & desarrollo , Luz , Solanum lycopersicum/metabolismo , Solanum lycopersicum/crecimiento & desarrollo , Plantones/crecimiento & desarrollo , Plantones/metabolismoRESUMEN
BACKGROUND: Recently emerging approaches to high-throughput plant phenotyping have discovered their importance as tools in unravelling the complex questions of plant growth, development and response to the environment, both in basic and applied science. High-throughput methods have been also used to study plant responses to various types of biotic and abiotic stresses (drought, heat, salinity, nutrient-starving, UV light) but only rarely to cold tolerance. RESULTS: We present here an experimental procedure of integrative high-throughput in-house phenotyping of plant shoots employing automated simultaneous analyses of shoot biomass and photosystem II efficiency to study the cold tolerance of pea (Pisum sativum L.). For this purpose, we developed new software for automatic RGB image analysis, evaluated various parameters of chlorophyll fluorescence obtained from kinetic chlorophyll fluorescence imaging, and performed an experiment in which the growth and photosynthetic activity of two different pea cultivars were followed during cold acclimation. The data obtained from the automated RGB imaging were validated through correlation of pixel based shoot area with measurement of the shoot fresh weight. Further, data obtained from automated chlorophyll fluorescence imaging analysis were compared with chlorophyll fluorescence parameters measured by a non-imaging chlorophyll fluorometer. In both cases, high correlation was obtained, confirming the reliability of the procedure described. CONCLUSIONS: This study of the response of two pea cultivars to cold stress confirmed that our procedure may have important application, not only for selection of cold-sensitive/tolerant varieties of pea, but also for studies of plant cold-response strategies in general. The approach, provides a very broad tool for the morphological and physiological selection of parameters which correspond to shoot growth and the efficiency of photosystem II, and is thus applicable in studies of various plant species and crops.
RESUMEN
Dark-induced growth (skotomorphogenesis) is primarily characterized by rapid elongation of the hypocotyl. We have studied the role of abscisic acid (ABA) during the development of young tomato (Solanum lycopersicum L.) seedlings. We observed that ABA deficiency caused a reduction in hypocotyl growth at the level of cell elongation and that the growth in ABA-deficient plants could be improved by treatment with exogenous ABA, through which the plants show a concentration dependent response. In addition, ABA accumulated in dark-grown tomato seedlings that grew rapidly, whereas seedlings grown under blue light exhibited low growth rates and accumulated less ABA. We demonstrated that ABA promotes DNA endoreduplication by enhancing the expression of the genes encoding inhibitors of cyclin-dependent kinases SlKRP1 and SlKRP3 and by reducing cytokinin levels. These data were supported by the expression analysis of the genes which encode enzymes involved in ABA and CK metabolism. Our results show that ABA is essential for the process of hypocotyl elongation and that appropriate control of the endogenous level of ABA is required in order to drive the growth of etiolated seedlings.
Asunto(s)
Ácido Abscísico/metabolismo , Oscuridad , Endorreduplicación/efectos de la radiación , Hipocótilo/crecimiento & desarrollo , Hipocótilo/efectos de la radiación , Solanum lycopersicum/crecimiento & desarrollo , Solanum lycopersicum/efectos de la radiación , Quinasas Ciclina-Dependientes/antagonistas & inhibidores , Citocininas/biosíntesis , Citocininas/metabolismo , Germinación/efectos de los fármacos , Germinación/efectos de la radiación , Homeostasis/efectos de la radiación , Hipocótilo/citología , Hipocótilo/metabolismo , Solanum lycopersicum/citología , Solanum lycopersicum/metabolismo , Desarrollo de la Planta/efectos de los fármacos , Desarrollo de la Planta/efectos de la radiación , Inhibidores de Proteínas Quinasas/farmacologíaRESUMEN
Phosphoglucose isomerase (PGI) catalyzes the reversible isomerization of glucose-6-phosphate and fructose-6-phosphate. It is involved in glycolysis and in the regeneration of glucose-6-P molecules in the oxidative pentose phosphate pathway (OPPP). In chloroplasts of illuminated mesophyll cells PGI also connects the Calvin-Benson cycle with the starch biosynthetic pathway. In this work we isolated pgi1-3, a mutant totally lacking pPGI activity as a consequence of aberrant intron splicing of the pPGI encoding gene, PGI1. Starch content in pgi1-3 source leaves was ca. 10-15% of that of wild type (WT) leaves, which was similar to that of leaves of pgi1-2, a T-DNA insertion pPGI null mutant. Starch deficiency of pgi1 leaves could be reverted by the introduction of a sex1 null mutation impeding ß-amylolytic starch breakdown. Although previous studies showed that starch granules of pgi1-2 leaves are restricted to both bundle sheath cells adjacent to the mesophyll and stomata guard cells, microscopy analyses carried out in this work revealed the presence of starch granules in the chloroplasts of pgi1-2 and pgi1-3 mesophyll cells. RT-PCR analyses showed high expression levels of plastidic and extra-plastidic ß-amylase encoding genes in pgi1 leaves, which was accompanied by increased ß-amylase activity. Both pgi1-2 and pgi1-3 mutants displayed slow growth and reduced photosynthetic capacity phenotypes even under continuous light conditions. Metabolic analyses revealed that the adenylate energy charge and the NAD(P)H/NAD(P) ratios in pgi1 leaves were lower than those of WT leaves. These analyses also revealed that the content of plastidic 2-C-methyl-D-erythritol 4-phosphate (MEP)-pathway derived cytokinins (CKs) in pgi1 leaves were exceedingly lower than in WT leaves. Noteworthy, exogenous application of CKs largely reverted the low starch content phenotype of pgi1 leaves. The overall data show that pPGI is an important determinant of photosynthesis, energy status, growth and starch accumulation in mesophyll cells likely as a consequence of its involvement in the production of OPPP/glycolysis intermediates necessary for the synthesis of plastidic MEP-pathway derived hormones such as CKs.