Higher Levels Of Jasmonic Acid Research Articles

Endophytic Fungi-Mediated Defense Signaling in Maize: Unraveling the Role of WRKY36 in Regulating Immunity against Spodoptera frugiperda.

Seed priming with beneficial endophytic fungi is an emerging sustainable strategy for enhancing plant resistance against insect pests. This study examined the effects of Beauvaria bassiana Bb20091317 and Metarhizium rileyi MrCDTLJ1 fungal colonization on maize growth, defence signalling, benzoxazinoid levels and gene expression. The colonization did not adversely affect plant growth but reduced larval weights of Spodoptera frugiperda. Maize leaves treated with M. rileyi exhibited higher levels of jasmonic acid, jasmonoyl-Isoleucine, salicylic acid, and indole acetic acid compared to control. B. bassiana and M. rileyi accelerated phytohormone increase upon S. frugiperda herbivory. Gene expression analysis revealed modulation of benzoxazinoid biosynthesis genes. We further elucidated the immune regulatory role of the transcription factor zmWRKY36 using virus-induced gene silencing (VIGS) in maize. zmWRKY36 positively regulates maize immunity against S. frugiperda, likely by interacting with defense-related proteins. Transient overexpression of zmWRKY36 in tobacco-induced cell death, while silencing in maize reduced chitin-triggered reactive oxygen species burst, confirming its immune function. Overall, B. bassiana and M. rileyi successfully colonized maize, impacting larval growth, defense signalling, and zmWRKY36-mediated resistance. This sheds light on maize-endophyte-insect interactions for sustainable plant protection.

Transcription Factor and Zeatin Co-Regulate Mixed Catkin Differentiation of Chinese Chestnut (Castanea mollissima)

Castanea mollissima is an important monoecious fruit crop with high economic and ecological value in China. However, its yield is restricted by an imbalanced ratio of male and female flowers for chestnut production. To address this issue, we examined the morphology of bisexual flower organs, measured the levels of endogenous hormones in the flowers, profiled gene expression related to plant hormone biosynthesis and signaling pathways and transcription factors, and investigated the effects of exogenous jasmonic acid (JA) and zeatin (ZT) hormone application on flower development in C. mollissima ‘Tanqiao’. Morphological studies indicated that the development of male and female flowers can be divided into nine and eight stages, respectively. Male flowers contained higher levels of gibberellic acid (GA3) and abscisic acid (ABA) than female flowers, whereas female flowers had higher levels of JA and ZT. The analysis of the Kyoto encyclopedia of genes and genomes (KEGG) pathways revealed that the major significant enrichment pathways of differentially expressed genes (DEGs) consisted of plant hormone signal transduction and zeatin biosynthesis. Through time-series analyses, we screened 3 genes related to jasmonic acid biosynthesis and signal transduction and 21 genes related to zeatin biosynthesis and transduction. Among these genes, only the gene family LOG, related to zeatin biosynthesis, was highly expressed in female flowers. This result indicated that LOG may be the core gene hormone family involved in regulating female flower development. However, a weighted gene co-expression network analysis (WGCNA) suggested that IDD7 was the core gene involved in regulating female flower development. The results of exogenous hormone application indicated that zeatin could greatly increase the quantity of fertile female flowers, but JA was not significant. These findings demonstrated that zeatin and transcription factors were crucial regulators in the formation of female flowers in C. mollissima.

Hormone and carbohydrate regulation of defense secondary metabolites in a Mediterranean forest during drought

Plants have evolved complex physiological mechanisms to regulate production of defense secondary metabolites (SMs) to cope with environmental stress such as drought. Yet, these mechanisms remain under-studied in mature trees, limiting our ability to understand effects of climate change on tree productivity and survival. We investigated defense SMs and their relations to growth, nonstructural carbohydrates and phytohormones across seasons, in a Mediterranean forest dominated by two tree species with different resistance to drought. Our results showed seasonality of leaf SMs, with a strong accumulation of SMs during cold and dry conditions in winter. The drought-resistant species Phillyrea latifolia had high levels of SMs in both leaves (ca. 10–20%) and stem phloem (4–6%), but experimental drought decreased SMs and soluble sugars in stem phloem in spring and summer. By contrast, the drought-susceptible species Quercus ilex had relatively low and constant levels of SMs, irrespective of drought. We further showed that P. latifolia leaves had generally higher levels of jasmonic acid and lower salicylic acid than Q. ilex leaves, potentially leading to differences in SMs and growth between the two co-occucrring species. Our study suggests that species- and organ-specific dynamics of defense SMs are driven by phytohormones and carbohydrates in mature trees, and that P. latifolia may become less resistant to abiotic and biotic stress due to reduced phloem defense SMs and carbohydrates in a drying climate.

Revealing Differential Expression of Phytohormones in Sorghum in Response to Aphid Attack Using the Metabolomics Approach.

Sorghum (Sorghum bicolor) is an important multipurpose crop grown worldwide, but like many other crops, it is often threatened by insect pests. Sugarcane aphid (SCA, Melanaphis sacchari Zehntner), for example, is one of the most severe pests in sorghum, which causes plant damage and yield loss. The main objective of this study was to assess the effect of phytohormones on host plant resistance to aphid attack. Two sorghum genotypes, BTx623 (susceptible) and Tx2783 (resistant), were selected for a comparative analysis of differential expression of a group of phytohormones in response to aphid infestation. The quantification of phytohormones through LC-MS demonstrated higher levels of jasmonic acid (JA), salicylic acid (SA), abscisic acid (ABA), and auxins in the resistant genotype infested with SCA. The PCA plot supports the strong differential responses between resistant and susceptible genotypes, indicating a positive correlation between JA and ABA and a negative correlation between SA and auxins. Similarly, RT-PCR results of the phytohormones-related marker genes showed higher expression in the resistant genotype compared to the susceptible one. Furthermore, to corroborate the role of phytohormones in plant defense, the susceptible genotype was treated with SA, JA, and ABA. The exogenous application of SA and JA + ABA significantly reduced plant mortality, aphid number, and damage in the susceptible genotype, suggesting a strong correlation between phytohormones and plant survival. Our findings indicate that phytohormones play positive roles in plant defense against aphids and provide new insights into the molecular mechanisms operating in plants for self-protection. These findings could also stimulate further research into the mystery about the regulation of phytohormone production during plant interaction with aphids.

Jasmonate regulates plant resistance to Pectobacterium brasiliense by inducing indole glucosinolate biosynthesis.

Pectobacterium brasiliense (P. brasiliense) is a necrotrophic bacterium that causes the soft rot disease in Brassica rapa. However, the mechanisms underlying plant immune responses against necrotrophic bacterial pathogens with a broad host range are still not well understood. Using a flg22-triggered seedling growth inhibition (SGI) assay with 455 Brassica rapa inbred lines, we selected six B. rapa flagellin-insensitive lines (Brfin2-7) and three B. rapa flagellin-sensitive lines (Brfs1-3). Brfin lines showed compromised flg22-induced immune responses (oxidative burst, mitogen-activated protein kinase (MAPK) activation, and seedling growth inhibition) compared to the control line R-o-18; nevertheless, they were resistant to P. brasiliense. To explain this, we analyzed the phytohormone content and found that most Brfin lines had higher P. brasiliense-induced jasmonic acid (JA) than Brfs lines. Moreover, MeJA pretreatment enhanced the resistance of B. rapa to P. brasiliense. To explain the correlation between the resistance of Brfin lines to P. brasiliense and activated JA signaling, we analyzed pathogen-induced glucosinolate (GS) content in B. rapa. Notably, in Brfin7, the neoglucobrassicin (NGBS) content among indole glucosinolates (IGS) was significantly higher than that in Brfs2 following P. brasiliense inoculation, and genes involved in IGSs biosynthesis were also highly expressed. Furthermore, almost all Brfin lines with high JA levels and resistance to P. brasiliense had higher P. brasiliense-induced NGBS levels than Brfs lines. Thus, our results show that activated JA-mediated signaling attenuates flg22-triggered immunity but enhances resistance to P. brasiliense by inducing indole glucosinolate biosynthesis in Brassica rapa. This study provides novel insights into the role of JA-mediated defense against necrotrophic bacterial pathogens within a broad host range.

Identification of Robinia pseudoacacia target proteins responsive to Mesorhizobium amphore CCNWGS0123 effector protein NopT.

Nodulation outer proteins secreted via type 3 secretion systems are involved in the process of symbiosis between legume plants and rhizobia. To study the function of NopT in symbiosis, we mutated nopT in Mesorhizobium amphore CCNWGS0123 (GS0123), which can nodulate black locust (Robinia pseudoacacia). The nopT mutant induced higher levels of jasmonic acid, salicylic acid, and hydrogen peroxide accumulation in the roots of R. pseudoacacia compared with wild-type GS0123. The ΔnopT mutant induced higher disease-resistant gene expression 72 hours post-inoculation (hpi), whereas GS0123 induced higher disease-resistant gene expression earlier, at 36 hpi. Compared with the nopT mutant, GS0123 induced the up-regulation of most genes at 36 hpi and the down-regulation of most genes at 72 hpi. Proteolytically active NopT_GS0123 induced hypersensitive responses when expressed transiently in tobacco leaves (Nicotiana benthamiana). Two NopT_GS0123 targets in R. pseudoacacia were identified, ATP-citrate synthase alpha chain protein 2 and hypersensitive-induced response protein. Their interactions with NopT_GS0123 triggered resistance by the plant immune system. In conclusion, NopT_GS0123 inhibited the host plant immune system and had minimal effect on nodulation in R. pseudoacacia. Our results reveal the underlying molecular mechanism of NopT function in plant-symbiont interactions.

The Lamin-Like LITTLE NUCLEI 1 (LINC1) Regulates Pattern-Triggered Immunity and Jasmonic Acid Signaling.

Pathogen-associated molecular pattern (PAMP) recognition occurs by plasma membrane located receptors that induce among other processes nuclear gene expression. However, signaling to the nuclear compartment is restricted by the nuclear envelope and nuclear pore complexes. We show here that among the four Arabidopsis lamin homologs LITTLE NUCLEI/CROWDED NUCLEI (LINC/CRWN), LINC1 plays an important role in PTI and jasmonic acid (JA) signaling. We show that linc1 knock out mutants affect PAMP-triggered MAPK activation and growth inhibition, but not reactive oxygen species or callose accumulation. We also demonstrate that linc1 mutants are compromised in regulating PAMP-triggered pathogen-related genes, in particular encoding factors involved in JA signaling and responses. Expression of a number of JAZ domain proteins, the key JA-related transcription factor MYC2 as well as key MYB transcription factors and biosynthesis genes of both the indole and aliphatic glucosinolate pathways are changed in linc1 mutants. Moreover, PAMP triggers JA and JA-Ile accumulation in linc1 mutants, whereas salicylic acid levels are unchanged. Despite impairment in PAMP-triggered immunity, linc1 mutants still show basal immunity towards Pseudomonas syringae DC3000 strains. High JA levels usually render plants resistant to necrotrophic pathogen. Thus, linc1 mutants show enhanced resistance to Botrytis cinerea infection. In accordance with a general role of LINC1 in JA signaling, linc1 mutants are hypersensitive to growth inhibition to external JA. In summary, our findings show that the lamin-like LINC1 protein plays a key role in JA signaling and regulation of PTI responses in Arabidopsis.

In vitro evaluation of berries of various Vitis genotypes for disease resistance to Botrytis cinerea

Berries of 41 Vitis genotypes were evaluated for resistance to B. cinerea. Evaluation revealed that four genotypes were highly resistant (HR), eight resistant (R), eighteen susceptible (S) and eleven highly susceptible (HS). We further evaluated HR genotype 'Dong fang zhi xing' and HS genotype 'Gold finger' by comparing the fungal growth, reactive oxygen species (ROS) responses, jasmonic acid (JA) levels, anti-oxidants, e.g., Peroxidase (POD), Superoxide dismutase (SOD) and Malondialhydrate (MDA) content changes after infection with B. cinerea. Our results confirmed that the elevated resistance of 'Dong fang zhi xing' was due to weak fungal development, low ROS production, timely elevation of anti-oxidative functions, and high JA levels. Moreover, HS 'Gold finger' infection was severe and sustained ROS production which may be due to its relatively unchanged anti-oxidative activities and low JA level. Our results could help grape breeders to select suitable germplasm for future research work.

Mutation in the Arabidopsis regulatory-associated protein TOR 1B (RAPTOR1B) leads to decreased jasmonates levels in leaf tissue

ABSTRACTThe Target of Rapamycin (TOR) complex (TORC) regulates plant growth and development by modulation of metabolism in response to environmental cues. TORC contains in its core the TOR kinase and two interacting partners, namely; regulatory-associated partner of TOR (RAPTOR) and lethal with sec thirteen protein 8 (LST8). RAPTOR is described to act as a scaffold protein which recruits substrates for phosphorylation to the TOR kinase.In the current manuscript we show that mutation of Arabidopsis RAPTOR1B leads to significantly decreased levels of free jasmonic acid (JA), jasmonoyl-(L)-isoleucine (JA-Ile) as well as its biosynthetic precursor 12-oxo-phytodienoic acid (OPDA). Although raptor1b leaves showed decreased basic JA level compared to WT, the mutant responded substantially to wounding stress by producing the same amount of JA as WT. Furthermore, we could show that the chemical inhibition of TOR by AZD-8055 led to an opposite response. AZD-treated WT and raptor1b leaves accumulated high JA levels. These results strongly imply that the TOR signaling pathway is responding differentially to the inhibition of the TOR kinase as compared to the inhibition of the scaffold protein RAPTOR.

Herbivore-induced volatile emission from old-growth black poplar trees under field conditions

Herbivory is well known to trigger increased emission of volatile organic compounds (VOCs) from plants, but we know little about the responses of mature trees. We measured the volatiles emitted by leaves of old-growth black poplar (Populus nigra) trees after experimental damage by gypsy moth (Lymantria dispar) caterpillars in a floodplain forest, and studied the effect of herbivory on the transcript abundance of two genes involved in the biosynthesis of VOCs, and the accumulation of defence phytohormones. Herbivory significantly increased volatile emission from the experimentally damaged foliage, but not from adjacent undamaged leaves in the damaged branches (i.e., no systemic response). Methylbutyraldoximes, 4,8-dimethyl-1,3,7-nonatriene (DMNT), (Z)-3-hexenol and (E)-β-ocimene, amongst other compounds, were found to be important in distinguishing the blend of herbivore-damaged vs. undamaged leaves. Herbivory also increased expression of PnTPS3 (described here for the first time) and PnCYP79D6-v4 genes at the damaged sites, these genes encode for an (E)-β-ocimene synthase and a P450 enzyme involved in aldoxime formation, respectively, demonstrating de novo biosynthesis of the volatiles produced. Herbivore-damaged leaves had significantly higher levels of jasmonic acid and its conjugate (−)-jasmonic acid-isoleucine. This study shows that mature trees in the field have a robust response to herbivory, producing induced volatiles at the damaged sites even after previous natural herbivory and under changing environmental conditions, however, further studies are needed to establish whether the observed absence of systemic responses is typical of mature poplar trees or if specific conditions are required for their induction.

Screening Vitis Genotypes for Responses to Botrytis cinerea and Evaluation of Antioxidant Enzymes, Reactive Oxygen Species and Jasmonic Acid in Resistant and Susceptible Hosts.

Botrytis cinerea is a necrotrophic fungal phytopathogen with devastating effects on many Vitis genotypes. Here, a screening of 81 Vitis genotypes for leaf resistance to B. cinerea revealed two highly resistant (HR), twelve resistant (R), twenty-five susceptible (S) and forty-two highly susceptible (HS) genotypes. We focused on the HR genotype, ‘Zi Qiu’ (Vitis davidii), and the HS genotype ‘Riesling’ (V. vinifera), to elucidate mechanisms of host resistance and susceptibility against B. cinerea, using detached leaf assays. These involved a comparison of fungal growth, reactive oxygen species (ROS) responses, jasmonic acid (JA) levels, and changes in the anti-oxidative system between the two genotypes after inoculation with B. cinerea. Our results indicated that the high-level resistance of ‘Zi Qiu’ can be attributed to insignificant fungal development, low ROS production, timely elevation of anti-oxidative functions, and high JA levels. Moreover, severe fungal infection of ‘Riesling’ and sustained ROS production coincided with relatively unchanged anti-oxidative activity, as well as low JA levels. This study provides insights into B. cinerea infection in grape, which can be valuable for breeders by providing information for selecting suitable germplasm with enhanced disease resistance.

Biochar amendment changes jasmonic acid levels in two rice varieties and alters their resistance to herbivory

Biochar addition to soil not only sequesters carbon for the long-term but enhances agricultural productivity. Several well-known benefits arise from biochar amendment, including constant provision of nutrients, increased soil moisture retention, decreased soil bulk density, and sometimes the induction of systemic resistance against foliar and soil borne plant pathogens. However, no research has investigated the potential of biochar to increase resistance against herbivory. The white-backed plant hopper (WBPH) (Sogatella furcifera Horváth) is a serious agricultural pest that targets rice (Oryza sativa L.), a staple crop that feeds half of the world’s human population. Therefore, we investigated the (1) optimization of biochar amendment levels for two rice varieties (‘Cheongcheong’ and ‘Nagdong’) and (2) subsequent effects of different biochar amendments on resistance and susceptibility of these two varieties to WBPH infestation. Initial screening results for the optimization level revealed that the application of biochar 10% (w/w) to the rooting media significantly improved plant physiological characteristics of both rice varieties. However, levels of biochar amendment, mainly 1, 2, 3, and 20%, resulted in negative effects on plant growth characteristics. Cheongcheong and Nagdong rice plants grown with the optimum biochar level showed contrasting reactions to WBPH infestation. Specifically, biochar application significantly increased plant growth characteristics of Nagdong when exposed to WBPH infestation and significantly decreased these characteristics in Cheongcheong. The amount of WBPH-induced damage to plants was significantly lower and higher in Nagdong and Cheongcheong, respectively, compared to that in the controls. Higher levels of jasmonic acid caused by the biochar priming effect could have accumulated in response to WBPH infestation, resulting in a maladaptive response to stress, negatively affecting growth and resistance to WBPH in Cheongcheong. This study highlights the importance of investigating the effects of biochar on different rice varieties before application on a commercial scale to avoid potential crop losses.

Herbivore defense responses and associated herbivore defense mechanism as revealed by comparing a resistant wild soybean with a susceptible cultivar

Plants have evolved sophisticated defense mechanisms against herbivores to help them adapt to the environment. Understanding the defense mechanisms in plants can help us control insects in a more effective manner. In this study, we found that compared with Tianlong 2 (a cultivated soybean with insect susceptibility), ED059 (a wild soybean line with insect resistance) contains sharper pubescence tips, as well as lower transcript levels of wound-induced protein kinase (WIPK) and salicylic acid-induced protein kinase (SIPK), which are important mitogen-activated protein kinases involved in early defense response to herbivores. The observed lower transcript levels of WIPK and SIPK induced higher levels of jasmonic acid (JA), JA biosynthesis enzymes (AOC3) and some secondary metabolites in ED059. Functional analysis of the KTI1 gene via Agrobacterium-mediated transformation in Arabidopsis thaliana indicated that it plays an important role in herbivore defense in ED059. We further investigated the molecular response of third-instar Helicoverpa armigera (Hübner) larvae to Tianlong 2 and ED059. We found apoptotic cells only in the midguts of larvae that fed on ED059. Compared with larvae reared on the susceptible cultivar Tianlong 2, transcript levels of catalase (CAT) and glutathione S-transferase (GST) were up-regulated, whereas those of CAR, CHSB, and TRY were down-regulated in larvae that fed on the highly resistant variety ED059. We propose that these differences underlie the different herbivore defense responses of ED059 and Tianlong 2.

Drought stress affects plant metabolites and herbivore preference but not host location by its parasitoids.

One of the main abiotic stresses that strongly affects plant survival and the primary cause of crop loss around the world is drought. Drought stress leads to sequential morphological, physiological, biochemical and molecular changes that can have severe effects on plant growth, development and productivity. As a consequence of these changes, the interaction between plants and insects can be altered. Using cultivated Brassica oleracea plants, the parasitoid Microplitis mediator and its herbivorous host Mamestra brassicae, we studied the effect of drought stress on (1) the emission of plant volatile organic compounds (VOCs), (2) plant hormone titres, (3) preference and performance of the herbivore, and (4) preference of the parasitoid. Higher levels of jasmonic acid (JA) and abscisic acid (ABA) were recorded in response to herbivory, but no significant differences were observed for salicylic acid (SA) and indole-3-acetic acid (IAA). Drought significantly impacted SA level and showed a significant interactive effect with herbivory for IAA levels. A total of 55 VOCs were recorded and the difference among the treatments was influenced largely by herbivory, where the emission rate of fatty acid-derived volatiles, nitriles and (E)-4,8-dimethylnona-1,3,7-triene [(E)-DMNT] was enhanced. Mamestra brassicae moths preferred to lay eggs on drought-stressed over control plants; their offspring performed similarly on plants of both treatments. VOCs due to drought did not affect the choice of M. mediator parasitoids. Overall, our study reveals an influence of drought on plant chemistry and insect-plant interactions.

Arbuscular mycorrhiza increase artemisinin accumulation in Artemisia annua by higher expression of key biosynthesis genes via enhanced jasmonic acid levels.

It is becoming increasingly evident that the formation of arbuscular mycorrhiza (AM) enhances secondary metabolite production in shoots. Despite mounting evidence, relatively little is known about the underlying mechanisms. This study suggests that increase in artemisinin concentration in Artemisia annua colonized by Rhizophagus intraradices is due to altered trichome density as well as transcriptional patterns that are mediated via enhanced jasmonic acid (JA) levels. Mycorrhizal (M) plants had higher JA levels in leaf tissue that may be due to induction of an allene oxidase synthase gene (AOS), encoding one of the key enzymes for JA production. Non-mycorrhizal (NM) plants were exogenously supplied with a range of methyl jasmonic acid concentrations. When leaves of NM and M plants with similar levels of endogenous JA were compared, these matched closely in terms of shoot trichome density, artemisinin concentration, and transcript profile of artemisinin biosynthesis genes. Mycorrhization increased artemisinin levels by increasing glandular trichome density and transcriptional activation of artemisinin biosynthesis genes. Transcriptional analysis of some rate-limiting enzymes of mevalonate and methyl erythritol phosphate (MEP) pathways revealed that AM increases isoprenoids by induction of the MEP pathway. A decline in artemisinin concentration in shoots of NM and M plants treated with ibuprofen (an inhibitor of JA biosynthesis) further confirmed the implication of JA in the mechanism of artemisinin production.

Signals induced by exogenous nitric oxide and their role in controlling brown rot disease caused by Monilinia fructicola in postharvest peach fruit

Recent evidence suggests that nitric oxide (NO) signaling plays an important role in plant–pathogen interactions and that aconitase is a major target of NO. In the present study on the signaling role of NO in the elicitation of defense responses in peach fruit against Monilinia fructicola and subsequent effect on brown rot disease, 15 μM NO solution induced disease resistance in harvested peaches. As a potentiated elicitor, NO induced high levels of endogenous NO and superoxide (O2 −), hydrogen peroxide (H2O2), and NADPH oxidase and Ca2+-ATPase activity in the fruit. Aconitase activity in peach fruit was inhibited by NO. Activity of partially purified aconitase was inhibited in vitro by sodium nitroprusside (SNP) and H2O2; however, the inhibition could be relieved by carboxy-2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (cPTIO) or catalase (CAT), indicating that the defense response and signals induced by NO transduction depend on aconitase and conditions leading to elevated levels of NO; otherwise, H2O2 would inactivate aconitase directly in fruit. Treatment with NO resulted in salicylic acid (SA) accumulating during storage. Higher levels of jasmonic acid (JA) were detected in NO-treated fruit 48 h after the treatment. But after NO was removed, the level of SA and JA were lower than in the control. The results suggest that exogenous NO enhances resistance of harvested peach fruit against the fungus by inducing signals such as endogenous NO, reactive oxygen species (ROS), SA and JA and by inhibiting aconitase activity.

Laboratory testing and molecular analysis of the resistance of wild and cultivated soybeans to cotton bollworm, Helicoverpa armigera (Hübner)

Identifying a superior soybean variety with high defoliator resistance is important to avoid yield loss. Cotton bollworm (Helicoverpa armigera Hübner) is one of the major defoliators of soybean (Glycine max [L.] Merr.) worldwide. In this study, we evaluated the effect of H. armigera larvae on ED059, a wild soybean (Glycine soja Sieb. et Zucc.), and three cultivated soybean varieties: Tianlong 2, PI 535807, and PI 533604, in choice and no-choice assays. The percentage of ED059 leaflets consumed by H. armigera was lower than that of the three cultivated soybeans. Larvae that fed on ED059 exhibited low weight gain and high mortality rate. Waldbauer nutritional indices suggested that ED059 reduced the growth, consumption, and frass production of H. armigera larvae. Larvae that fed on ED059 showed lower efficiency of conversion of ingested and of digested food than those that fed on Tianlong 2 and PI 533604. However, they showed statistically similar consumption index and approximate digestibility compared with those fed on the three cultivated soybeans. Quantitative real-time PCR analysis revealed that 24h after insect attack, ED059 had higher transcript levels of Kunitz trypsin inhibitor 3, Cysteine proteinase inhibitor 2, and Nerolidol synthase 1 but a lower transcript level of Pathogenesis-related protein 1 than Tianlong 2. The gene expression results were consistent with the presence of higher levels of jasmonic acid (JA) and transcript levels of the JA biosynthesis enzyme allene oxide cyclase 3 in ED059 than in Tianlong 2. Our findings indicate that ED059 is a superior soybean line with strong insect resistance that may be mediated via the JA pathway.

The Hordeum vulgare signalling protein MAP kinase 4 is a regulator of biotic and abiotic stress responses

Mitogen activated protein kinase (MAP kinase) signal transduction pathways are important eukaryotic mechanisms for regulating cellular responses to stress. The objective of this work was to investigate the role of the barley MAP kinase HvMPK4 (a homologue of the Arabidopsis MAP kinase AtMPK1) in the plant response to biotic and abiotic stress. Transgenic barley plants bearing antisense or overexpression constructs for HvMPK4 were produced, and RNA blot analysis showed that HvMPK4 gene expression was much reduced in the antisense lines and approximately double in the overexpression lines. Three independent lines of each construct were tested for their response to a fungal pathogen and to salt treatment. The antisense lines were more resistant to the hemibiotrophic fungal pathogen Magnaporthe grisea, and showed enhanced levels of salicylic acid (SA) and of hydrogen peroxide following infection; HvMPK4 is thus a negative regulator of SA production post infection. The overexpression lines had constitutively higher levels of jasmonic acid and enhanced levels of ethylene following infection but were not more resistant to the pathogen. However the overexpression lines showed greater tolerance to abiotic stress, as following 2 weeks of salt treatment these lines showed less reduction in fresh and dry weight, accumulated less salt in the leaves and contained enhanced levels of the osmoprotectant amino acid, proline.

High levels of jasmonic acid antagonize the biosynthesis of gibberellins and inhibit the growth of Nicotiana attenuata stems

Hormones play pivotal roles in regulating plant development, growth, and stress responses, and cross-talk among different hormones fine-tunes various aspects of plant physiology. Jasmonic acid (JA) is important for plant defense against herbivores and necrotic fungi and also regulates flower development; in addition, Arabidopsis mutants over-producing JA usually have stunted stems and wound-induced jasmonates suppress Arabidopsis growth, suggesting that JA is also involved in stem elongation. Gibberellins (GAs) promote stem and leaf growth and modulate seed germination, flowering time, and the development of flowers, fruits, and seeds. However, little is known about the interaction between the JA and GA pathways. Two calcium-dependent protein kinases, CDPK4 and CDPK5, are important suppressors of JA accumulation in a wild tobacco species, Nicotiana attenuata. The stems of N.attenuata silenced in CDPK4 and CDPK5 (irCDPK4/5 plants) had dramatically increased levels of JA and exhibited stunted elongation and had very high contents of secondary metabolites. Genetic analysis indicated that the high JA levels in irCDPK4/5 stems accounted for the suppressed stem elongation and the accumulation of secondary metabolites. Supplementation of GA(3) to irCDPK4/5 plants largely restored normal stem growth to wild-type levels. Measures of GA levels indicated that over-accumulation of JA in irCDPK4/5 stems inhibited the biosynthesis of GAs. Finally, we show that JA antagonizes GA biosynthesis by strongly inhibiting the transcript accumulation of GA20ox and possibly GA13ox, the key genes in GA production, demonstrating that high JA levels antagonize GA biosynthesis in stems.

Transcriptomic analysis of oxylipin biosynthesis genes and chemical profiling reveal an early induction of jasmonates in chickpea roots under drought stress

Drought is one of the major constraints in subtropical agriculture. Therefore improving water stress tolerance is of great importance to breed for drought tolerance in future. The first plant organ sensing dehydration is the root. Aim of the present work was to clarify the potential impact of the phyto-oxylipins pathway on drought tolerance of chickpea (Cicer arietinum), the third important legume crop worldwide. Therefore, we measured the expression of key genes involved in oxylipins metabolism by qPCR on samples from stressed and non-stressed roots of a drought-tolerant and a drought-sensitive chickpea variety using commercially available TaqMan assays. We demonstrate that the drought tolerant variety reacts to drought with sustained and earlier activation of a specific lipoxygenase (Mt-LOX 1) gene, two hydroperoxide lyases (Mt-HPL 1 and Mt-HPL 2), an allene oxide synthase (Mt-AOS), and an oxo-phytodienoate reductase (Mt-OPR). We further show that gene over-expression positively correlates with the levels of major oxylipin metabolites from the AOS branch of the pathway, which finally leads to the synthesis of jasmonates. Higher levels of jasmonic acid (JA), its precursor 12-oxophytodienoic acid (OPDA) and the active form JA-isoleucine (JA-Ile) were especially detected in the root tissues of the tolerant variety, prompting us to assume a role of jasmonates in the early signalling of drought stress in chickpea and its involvement in the tolerance mechanism of the drought-tolerant variety.