Leaf Metabolism Research Articles

Bacillus amyloliquefaciens MZ895491: an endophytic bacterium as potential biocontrol agent in integrated pest management of fall armyworm in maize

ABSTRACT The fall armyworm, Spodoptera frugiperda is a major pest of maize crops and necessitates effective control measures. Excessive use of chemical pesticides often results in the development of resistant insect populations and environmental contamination, highlighting the need for environmentally friendly substitutes. This research aimed to estimate the potential of maize (COH6) apoplastic fluid bacterium Bacillus amyloliquefaciens (MZ895491), as a biocontrol agent against second instar S. frugiperda larvae under laboratory and pot culture conditions. The results suggest that the presence of B. amyloliquefaciens in the artificial diet could reduce the growth and development of S. frugiperda larvae. Metabolic profiling of extracellular components of B. amyloliquefaciens revealed the presence of numerous defensive and anti-feeding metabolites. A study conducted in pots demonstrated that foliar application of B. amyloliquefaciens to 5 d old maize resulted in a significant increase in antioxidant activity (83.3%), chlorophyll content (63.3%) and phenolic content (81.3%) in the leaves of 35 d old maize after 24 h of S. frugiperda infestation. Feeding bioassay further indicated significant reduction in the feeding capacity of S. frugiperda larvae on maize inoculated with B. amyloliquefaciens. The study observed that a metabolic modification in maize leaves likely led to reduced growth in larvae during the whole plant bioassay. This suggests that foliar application of B. amyloliquefaciens may effectively serve as one of the biocontrol agents against Spodoptera frugiperda. The application appears to alter the feeding behaviour of the larvae and modify maize leaf metabolism.

Constitutive and inducible tomato defenses contribute to Bacillus thuringiensis lethality against Spodoptera exigua

In nature, insect herbivory exerts continuous selective pressure on plants that, in turn, have developed a wide array of constitutive and inducible defenses to fight against attackers. Since plant defenses may affect higher trophic levels, including entomopathogens, further research is required to understand how plant compounds influence insect-pathogens interactions and their implications for integrated pest management programs. Here, we evaluated the impact of tomato defenses on the lethality produced by the bacterial entomopathogen Bacillus thuringiensis (Bt) against second instar larvae of Spodoptera exigua. We first examined the effect of constitutive defenses from fifteen wild tomato species and forty cultivated varieties (Solanum lycopersicum) on S. exigua larval growth and susceptibility to Bt. The results showed larvae fed on wild tomato species had a reduced larval growth compared to larvae fed on cultivated varieties, whereas susceptibility to Bt was similar between both groups. We then selected six cultivated varieties, including those with high and low impacts on larval growth and Bt-induced mortality, to further explore the effect of inducible defenses. Elicitation of defenses by methyl jasmonate (MeJA) reduced larval growth and increased basal mortality. Additionally, when larvae were infected with Bt, MeJA treatment further increased their susceptibility to the entomopathogen. Metabolomic analysis confirmed a reprogramming of tomato leaf metabolism following MeJA elicitation, with an induced accumulation of bioactive compounds, such as saponins or flavonoids, known for their anti-herbivory properties in most tomato varieties. Overall, these data reveal that both constitutive and inducible tomato defenses not only protect the plant by directly affecting the insect pest but also enhance the efficacy of bacterial entomopathogens

Unveiling the role of epigenetics in leaf senescence: a comparative study to identify different epigenetic regulations of senescence types in barley leaves

BackgroundDevelopmental leaf senescence (DLS) is an irreversible process followed by cell death. Dark-induced leaf senescence (DILS) is a reversible process that allows adaptations to changing environmental conditions. As a result of exposure to adverse environmental changes, plants have developed mechanisms that enable them to survive. One of these is the redirection of metabolism into the senescence pathway. The plant seeks to optimise resource allocation. Our research aims to demonstrate how epigenetic machinery regulates leaf senescence, including its irreversibility.ResultsIn silico analyses allowed the complex identification and characterisation of 117 genes involved in epigenetic processes in barley. These genes include those responsible for DNA methylation, post-translational histone modifications, and ATP-dependent chromatin remodelling complexes. We then performed RNAseq analysis after DILS and DLS to evaluate their expression in senescence-dependent leaf metabolism. Principal component analysis revealed that evaluated gene expression in developmental senescence was similar to controls, while induced senescence displayed a distinct profile. Western blot experiments revealed that senescence engages senescence-specific histone modification. During DILS and DLS, the methylation of histone proteins H3K4me3 and H3K9me2 increased. H3K9ac acetylation levels significantly decreased during DILS and remained unchanged during DLS.ConclusionsThe study identified different epigenetic regulations of senescence types in barley leaves. These findings are valuable for exploring epigenetic regulation of senescence-related molecular mechanisms, particularly in response to premature, induced leaf senescence. Based on the results, we suggest the presence of an epigenetically regulated molecular switch between cell survival and cell death in DILS, highlighting an epigenetically driven cell survival metabolic response.

Drought response modelling of leaf photosynthetic parameters in two Gossypium species

AbstractCotton is well adapted to dry areas, but progressive water deficits can lead to declines in net photosynthesis (A), ultimately reducing yield. However, the exact mechanism responsible for this decline in net photosynthesis (stomatal or non‐stomatal) is not fully understood under field conditions, partially due to limitations in the ability to collect critical data. To our knowledge, no other study has compared the drought responses of Pima and upland cotton using both CO2 response and chlorophyll fluorescence under field conditions. To this end, a field study was conducted to quantify the impact of progressive mild drought, as measured by midday stomatal conductance to water vapour (gs), on cotton leaf metabolism in Pima and upland cotton. Survey gas exchange and rapid photosynthetic CO2 response (RACiR) were conducted during flowering on the same leaf. The study observed decline in A as gs declined for both species. Correlation analysis indicated typical relationships with A and parameters associated with stomatal limitations such as decreased CO2 inside the leaf and at the site of carboxylation; however, it was found that while Pima exhibited a strong relationship between maximum electron transport rate (Jmax) and electron transport rate (ETR), upland cotton did not. Furthermore, when ETR is broken down into proportions contributing to net photosynthesis and photorespiration (ETRA, ETRP, respectively), we found that a greater proportion of ETR is being shuttled to the photorespiratory pathway in upland, relative to Pima as gs decreases. Our results fill critical knowledge gaps that can be useful for modellers and breeders when preparing for future climate change scenarios.

Growth conditions trigger genotype-specific metabolic responses that affect the nutritional quality of kale cultivars.

Kales (Brassica oleracea convar acephala) are fast-growing, nutritious leafy vegetables ideal for year-round indoor farming. However, selection of best cultivars for growth under artificial lighting necessitates a deeper understanding of leaf metabolism in different kale types. Here we examined a curly leaved cultivar Half Tall and a lacinato type cultivar Black Magic under moderate growth light (130 µmol photons m-1s-1/22°C) and high light (800 µmol photons m-1s-1/26°C) conditions. These conditions induced genotype-dependent differences in nutritionally important metabolites, especially anthocyanins and glucosinolates (GSLs), in the kale cultivars. In the pale green Half Tall, growth under high light conditions did not induce changes in either pigmentation or total GSL content. In contrast, the purple pigmentation of Black Magic intensified due to increased anthocyanin accumulation. Black Magic showed reduced amounts of indole GSLs and increased amounts of aliphatic GSLs under high light conditions, with notable cultivar-specific adjustments in individual GSL species. Correlation analysis of metabolite profiles suggested cultivar-specific metabolic interplay between serine biosynthesis and the production of indole GSLs. RNA sequencing identified candidate genes encoding metabolic enzymes and regulatory components behind anthocyanin and GSL biosynthesis. These findings improve the understanding of leaf metabolism and its effects on the nutritional quality of kale cultivars.

Brown macro-seaweeds derived agro-biostimulant for Zea mays farming in saline conditions: Growth enhancement and optimum biochemical and ion feedback

Salinity negatively affects maize growth, a cereal crop and used as a vital source of food production. Seaweeds are macro-aquatic biocatalysts and might act as a promising biostimulant for conferring crop salinity resistance by providing essential nutrients and growth-promoting regulators. This study was designed for the first time to examine the effects of two commonly found seaweeds, Iyengaria stellata and Colpomenia sinuosa, on the growth of maize under 0, 100, and 200 mM NaCl application. Plants treated with both salinities reduced plant growth, but the application of seaweed I. stellata and C. sinuosa significantly increased growth parameters, especially plant biomass (fresh and dry), length (root and shoot), and total plant height than other treatments. In addition, higher growth under seaweed amends was supported by improved chlorophyll, carbohydrates, carotenoids, and polyphenols as well as lower electrolyte leakages to escape from oxidative stress. Plants treated with both algal species enhanced Na+ and K+, which is important for osmotic balance and precise ion homeostasis and could be effective for optimum leaf metabolism for growth resilience of maize plants under saline-stress conditions. This study emphasizes the potential of C. sinuosa and I. stellata as effective biostimulants. They offer a sustainable approach to cultivating climate-resilient maize that can thrive in salt-affected environments.

Genotype-Dependent Response of Root Microbiota and Leaf Metabolism in Olive Seedlings Subjected to Drought Stress.

Under stress or in optimum conditions, plants foster a specific guild of symbiotic microbes to strengthen pivotal functions including metabolic regulation. Despite that the role of the plant genotype in microbial selection is well documented, the potential of this genotype-specific microbial assembly in maintaining the host homeostasis remains insufficiently investigated. In this study, we aimed to assess the specificity of the foliar metabolic response of contrasting olive genotypes to microbial inoculation with wet-adapted consortia of plant-growth-promoting rhizobacteria (PGPR), to see if previously inoculated plants with indigenous or exogenous microbes would display any change in their leaf metabolome once being subjected to drought stress. Two Tunisian elite varieties, Chetoui (drought-sensitive) and Chemleli (drought-tolerant), were tested under controlled and stressed conditions. Leaf samples were analyzed by gas chromatography-mass spectrometry (GC-TOFMS) to identify untargeted metabolites. Root and soil samples were used to extract microbial genomic DNA destined for bacterial community profiling using 16S rRNA amplicon sequencing. Respectively, the score plot analysis, cluster analysis, heat map, Venn diagrams, and Krona charts were applied to metabolic and microbial data. Results demonstrated dynamic changes in the leaf metabolome of the Chetoui variety in both stress and inoculation conditions. Under the optimum state, the PGPR consortia induced noteworthy alterations in metabolic patterns of the sensitive variety, aligning with the phytochemistry observed in drought-tolerant cultivars. These variations involved fatty acids, tocopherols, phenols, methoxyphenols, stilbenoids, triterpenes, and sugars. On the other hand, the Chemleli variety displaying comparable metabolic profiles appeared unaffected by stress and inoculation probably owing to its tolerance capacity. The distribution of microbial species among treatments was distinctly uneven. The tested seedlings followed variety-specific strategies in selecting beneficial soil bacteria to alleviate stress. A highly abundant species of the wet-adapted inoculum was detected only under optimum conditions for both cultivars, which makes the moisture history of the plant genotype a selective driver shaping microbial community and thereby a useful tool to predict microbial activity in large ecosystems.

Differential effects of elevated CO2 on awn and glume metabolism in durum wheat (Triticum durum).

While the effect of CO2 enrichment on wheat (Triticum spp.) photosynthesis, nitrogen content or yield has been well-studied, the impact of elevated CO2 on metabolic pathways in organs other than leaves is poorly documented. In particular, glumes and awns, which may refix CO2 respired by developing grains and be naturally exposed to higher-than-ambient CO2 mole fraction, could show specific responses to elevated CO2 . Here, we took advantage of a free-air CO2 enrichment experiment and performed multilevel analyses, including metabolomics, ionomics, proteomics, major hormones and isotopes in Triticum durum . While in leaves, elevated CO2 tended to accelerate amino acid metabolism with many significantly affected metabolites, the effect on glumes and awns metabolites was modest. There was a lower content in compounds of the polyamine pathway (along with uracile and allantoin) under elevated CO2 , suggesting a change in secondary N metabolism. Also, cytokinin metabolism appeared to be significantly affected under elevated CO2 . Despite this, elevated CO2 did not affect the final composition of awn and glume organic matter, with the same content in carbon, nitrogen and other elements. We conclude that elevated CO2 mostly impacts on leaf metabolism but has little effect in awns and glumes, including their composition at maturity.

Reprogramming the translatome during daily light transitions as affected by cytosolic glyceraldehyde-3-phosphate dehydrogenases GAPC1/C2.

Dark-light and light-dark transitions during the day are switching points of leaf metabolism that strongly affect the regulatory state of the cells, and this change is hypothesized to affect the translatome. The cytosolic glyceraldehyde-3-phosphate dehydrogenases GAPC1 and GAPC2 function in glycolysis, and carbohydrate and energy metabolism, but GAPC1/C2 also shows moonlighting functions in gene expression and post-transcriptional regulation. In this study we examined the rapid reprogramming of the translatome that occurs within 10 min at the end of the night and the end of the day in wild-type (WT) Arabidopsis and a gapc1/c2 double-knockdown mutant. Metabolite profiling compared to the WT showed that gapc1/c2 knockdown led to increases in a set of metabolites at the start of day, particularly intermediates of the citric acid cycle and linked pathways. Differences in metabolite changes were also detected at the end of the day. Only small sets of transcripts changed in the total RNA pool; however, RNA-sequencing revealed major alterations in polysome-associated transcripts at the light-transition points. The most pronounced difference between the WT and gapc1/c2 was seen in the reorganization of the translatome at the start of the night. Our results are in line with the proposed hypothesis that GAPC1/C2 play a role in the control of the translatome during light/dark transitions.

Kinetic modeling identifies targets for engineering improved photosynthetic efficiency in potato (Solanum tuberosum cv. Solara).

Potato (Solanum tuberosum) is a significant non-grain food crop in terms of global production. However, its yield potential might be raised by identifying means to release bottlenecks within photosynthetic metabolism, from the capture of solar energy to the synthesis of carbohydrates. Recently, engineered increases in photosynthetic rates in other crops have been directly related to increased yield - how might such increases be achieved in potato? To answer this question, we derived the photosynthetic parameters Vcmax and Jmax to calibrate a kinetic model of leaf metabolism (e-Photosynthesis) for potato. This model was then used to simulate the impact of manipulating the expression of genes and their protein products on carbon assimilation rates in silico through optimizing resource investment among 23 photosynthetic enzymes, predicting increases in photosynthetic CO2 uptake of up to 67%. However, this number of manipulations would not be practical with current technologies. Given a limited practical number of manipulations, the optimization indicated that an increase in amounts of three enzymes - Rubisco, FBP aldolase, and SBPase - would increase net assimilation. Increasing these alone to the levels predicted necessary for optimization increased photosynthetic rate by 28% in potato.

Beyond photorespiration: the significance of glycine and serine in leaf metabolism.

The elucidation and removal of photorespiratory metabolic constraints will be necessary to improve crop yield in the next agricultural revolution. Fu et al. studied metabolic fluxes in the photorespiratory pathway and report that serine is the major export, whereas dynamic alterations in glycine pools orchestrate CO2 assimilation during the induction and relaxation of photorespiration.

Daylength variation affects growth, photosynthesis, leaf metabolism, partitioning, and metabolic fluxes.

Daylength, a seasonal and latitudinal variable, exerts a substantial impact on plant growth. However, the relationship between daylength and growth is nonproportional, suggesting the existence of adaptive mechanisms. Thus, our study aimed to comprehensively investigate the adaptive strategies employed by plants in response to daylength variation. We grew false flax (Camelina sativa) plants, a model oilseed crop, under long-day (LD) and short-day (SD) conditions and used growth measurements, gas exchange measurements, and isotopic labeling techniques, including 13C, 14C, and 2H2O, to determine responses to different daylengths. Our findings revealed that daylength influences various growth parameters, photosynthetic physiology, carbon partitioning, metabolic fluxes, and metabolite levels. SD plants employed diverse mechanisms to compensate for reduced CO2 fixation in the shorter photoperiod. These mechanisms included enhanced photosynthetic rates and reduced respiration in the light (RL), leading to increased shoot investment. Additionally, SD plants exhibited reduced rates of the glucose 6-phosphate (G6P) shunt and greater partitioning of sugars into starch, thereby sustaining carbon availability during the longer night. Isotopic labeling results further demonstrated substantial alterations in the partitioning of amino acids and TCA cycle intermediates between rapidly and slowly turning over pools. Overall, the results point to multiple developmental, physiological, and metabolic ways in which plants adapt to different daylengths to maintain growth.

Limits of thermal and hydrological tolerance in a foundation tree species (Populus fremontii) in the desert southwestern United States.

Populus fremontii is among the most dominant, and ecologically important riparian tree species in the western United States and can thrive in hyper-arid riparian corridors. Yet, P. fremontii forests have rapidly declined over the last decade, particularly in places where temperatures sometimes exceed 50°C. We evaluated high temperature tolerance of leaf metabolism, leaf thermoregulation, and leaf hydraulic function in eight P. fremontii populations spanning a 5.3°C mean annual temperature gradient in a well-watered common garden, and at source locations throughout the lower Colorado River Basin. Two major results emerged. First, despite having an exceptionally high Tcrit (the temperature at which Photosystem II is disrupted) relative to other tree taxa, recent heat waves exceeded Tcrit , requiring evaporative leaf cooling to maintain leaf-to-air thermal safety margins. Second, in midsummer, genotypes from the warmest locations maintained lower midday leaf temperatures, a higher midday stomatal conductance, and maintained turgor pressure at lower water potentials than genotypes from more temperate locations. Taken together, results suggest that under well-watered conditions, P. fremontii can regulate leaf temperature below Tcrit along the warm edge of its distribution. Nevertheless, reduced Colorado River flows threaten to lower water tables below levels needed for evaporative cooling during episodic heat waves.

A 3-year application of different mycorrhiza-based plant biostimulants distinctively modulates photosynthetic performance, leaf metabolism, and fruit quality in grapes (Vitis vinifera L.).

The use of microbial biostimulants in agriculture is recognized as a sustainable approach to promoting crop productivity and quality due to improved nutrient uptake, enhanced stress tolerance, and improved ability to cope with non-optimal environments. The present paper aimed to comparatively investigate the effect of seven different commercial mycorrhizal-based treatments in terms of yield, phytochemical components, and technological traits of Malvasia di Candia Aromatica grape (Vitis vinifera L.) plants. Metabolomic analysis and photosynthetic performance were first investigated in leaves to point out biochemical differences related to plant growth. Higher photosynthetic efficiency and better PSII functioning were found in biostimulant-treated vines, reflecting an overall decrease in photoinhibition compared to untreated plants. Untargeted metabolomics followed by multivariate statistics highlighted a robust reprogramming of primary (lipids) and secondary (alkaloids and terpenoids) metabolites in treated plants. The analysis of berry yield and chemical components exhibited significant differences depending on the biostimulant product. Generally, berries obtained from treated plants displayed improved contents of polyphenols and sugars, while yield remained unchanged. These results elucidated the significant role of microbial biostimulants in determining the quality of grape berries and eliciting biochemical changes in vines.

Marine heatwave and reduced light scenarios cause species-specific metabolomic changes in seagrasses under ocean warming.

Climate change and extreme climatic events, such as marine heatwaves (MHWs), are threatening seagrass ecosystems. Metabolomics can be used to gain insight into early stress responses in seagrasses and help to develop targeted management and conservation measures. We used metabolomics to understand the temporal and mechanistic response of leaf metabolism in seagrasses to climate change. Two species, temperate Posidonia australis and tropical Halodule uninervis, were exposed to a combination of future warming, simulated MHW with subsequent recovery period, and light deprivation in a mesocosm experiment. The leaf metabolome of P. australis was altered under MHW exposure at ambient light while H. uninervis was unaffected. Light deprivation impacted both seagrasses, with combined effects of heat and low light causing greater alterations in leaf metabolism. There was no MHW recovery in P. australis. Conversely, the heat-resistant leaf metabolome of H. uninervis showed recovery of sugars and intermediates of the tricarboxylic acid cycle under combined heat and low light exposure, suggesting adaptive strategies to long-term light deprivation. Overall, this research highlights how metabolomics can be used to study the metabolic pathways of seagrasses, identifies early indicators of environmental stress and analyses the effects of environmental factors on plant metabolism and health.

Content of Chlorophyll, Antioxidants, and Metabolite Compounds in the Leaf Development Stage of Murraya koenigii (L.) Spreng

Murraya koenigii (L.) Spreng is often known as Temurui or Curry as a local name in Indonesia, and has long been used in medicine and has as a potential as a multi-medicinal plant. Temurui as a producer of bioactivity has the antioxidant properties. This is due to the presence of metabolite compounds contained in the leaves of the Temurui plant. Leaves are organs that are always growing where chemical components such as chlorophyll in the leaves will also develop thereby affecting leaf metabolism in plants. This study aims to determine the contens of chlorophyll, contens of antioxidants and metabolite compounds in Temurui leaves at the level of leaf development. Different leaf extractions are also used to measure the antioxidant, chlorophyll content and metabolite compounds found in Temurui leaves. The research results obtained on young, and old Temurui leaves obtained a total of 3 groups of dominant compounds, terpenoids, organic acids and esters, and mature leaves not find ester compound. The level of leaf development affects the contents of antioxidants and chlorophyll contents. Chlorophyll content in mature leaves was 44.60ug/ml, young leaves were 41.28ug/ml and old leaves were 30.27ug/ml while antioxidant contents in young leaves were 8.949ppm, mature leaves were 8.85ppm and old leaves were 8.429ppm.

Plant NADPH-dependent thioredoxin reductases are crucial for the metabolism of sink leaves and plant acclimation to elevated CO2.

Plants contain three NADPH-thioredoxin reductases (NTR) located in the cytosol/mitochondria (NTRA/B) and the plastid (NTRC) with important metabolic functions. However, mutants deficient in all NTRs remained to be investigated. Here, we generated and characterised the triple Arabidopsis ntrabc mutant alongside with ntrc single and ntrab double mutants under different environmental conditions. Both ntrc and ntrabc mutants showed reduced growth and substantial metabolic alterations, especially in sink leaves and under high CO2 (HC), as compared to the wild type. However, ntrabc showed higher effective quantum yield of PSII under both constant and fluctuating light conditions, altered redox states of NADH/NAD+ and glutathione (GSH/GSSG) and lower potential quantum yield of PSII in sink leaves in ambient but not high CO2 concentrations, as compared to ntrc, suggesting a functional interaction between chloroplastic and extra-chloroplastic NTRs in photosynthesis regulation depending on leaf development and environmental conditions. Our results unveil a previously unknown role of the NTR system in regulating sink leaf metabolism and plant acclimation to HC, while it is not affecting full plant development, indicating that the lack of the NTR system can be compensated, at least to some extent, by other redox mechanisms.

CO2 recycling by phosphoenolpyruvate carboxylase enables cassava leaf metabolism to tolerate low water availability.

Cassava is a staple crop that acclimatizes well to dry weather and limited water availability. The drought response mechanism of quick stomatal closure observed in cassava has no explicit link to the metabolism connecting its physiological response and yield. Here, a genome-scale metabolic model of cassava photosynthetic leaves (leaf-MeCBM) was constructed to study on the metabolic response to drought and stomatal closure. As demonstrated by leaf-MeCBM, leaf metabolism reinforced the physiological response by increasing the internal CO2 and then maintaining the normal operation of photosynthetic carbon fixation. We found that phosphoenolpyruvate carboxylase (PEPC) played a crucial role in the accumulation of the internal CO2 pool when the CO2 uptake rate was limited during stomatal closure. Based on the model simulation, PEPC mechanistically enhanced drought tolerance in cassava by providing sufficient CO2 for carbon fixation by RuBisCO, resulting in high production of sucrose in cassava leaves. The metabolic reprogramming decreased leaf biomass production, which may lead to maintaining intracellular water balance by reducing the overall leaf area. This study indicates the association of metabolic and physiological responses to enhance tolerance, growth, and production of cassava in drought conditions.

Identification of Photosynthesis Characteristics and Chlorophyll Metabolism in Leaves of Citrus Cultivar (Harumi) with Varying Degrees of Chlorosis.

In autumn and spring, citrus leaves with a Ponkan (Citrus reticulata Blanco cv. Ponkan) genetic background (Harumi, Daya, etc.) are prone to abnormal physiological chlorosis. The effects of different degrees of chlorosis (normal, mild, moderate and severe) on photosynthesis and the chlorophyll metabolism of leaves of Citrus cultivar (Harumi) were studied via field experiment. Compared with severe chlorotic leaves, the results showed that chlorosis could break leaf metabolism balance, including reduced chlorophyll content, photosynthetic parameters, antioxidant enzyme activity and enzyme activity related to chlorophyll synthesis, increased catalase and decreased enzyme activity. In addition, the content of chlorophyll synthesis precursors showed an overall downward trend expected for uroporphyrinogen III. Furthermore, the relative expression of genes for chlorophyll synthesis (HEMA1, HEME2, HEMG1 and CHLH) was down-regulated to some extent and chlorophyll degradation (CAO, CLH, PPH, PAO and SGR) showed the opposite trend with increased chlorosis. Changes in degradation were more significant. In general, the chlorosis of Harumi leaves might be related to the blocked transformation of uroporphyrinogen III (Urogen III) to coproporphyrinogen III (Coprogen III), the weakening of antioxidant enzyme system activity, the weakening of chlorophyll synthesis and the enhancement in degradation.

Knock out of amino acid transporter gene OsLHT1 accelerates leaf senescence and enhances resistance to rice blast fungus.

Plant amino acid transporters regulate not only long-distance transport and reallocation of nitrogen (N) from source to sink organs, but also the amount of amino acids in leaves hijacked by invading pathogens. However, the function of amino acid transporters in plant defense responses to pathogen infection remains unknown. In this study, we found that the rice amino acid transporter gene OsLHT1 was expressed in leaves and up-regulated by maturation, N starvation, and inoculation of the blast fungus Magnaporthe oryzae. Knock out of OsLHT1 resulted in development stage- and N supply-dependent premature senescence of leaves at the vegetative growth stage. In comparison with the wild type, Oslht1 mutant lines showed sustained rusty red spots on fully mature leaf blades irrespective of N supply levels. Notably, no relationship between the severity of leaf rusty red spots and concentration of total N or amino acids was found in Oslht1 mutants at different developmental stages. Disruption of OsLHT1 altered transport and metabolism of amino acids and biosynthesis of flavones and flavonoids, enhanced expression of jasmonic acid- and salicylic acid-related defense genes, production of jasmonic acid and salicylic acid, and accumulation of reactive oxygen species. OsLHT1 inactivation dramatically prevented the leaf invasion by M. oryzae, a hemi-biotrophic ascomycete fungus. Overall, these results establish a link connecting the activity of an amino acid transporter with leaf metabolism and defense against rice blast fungus.