Conventional and unconventional ubiquitination in plant immunity

Ubiquitination is one of the most abundant types of protein post‐translational modification (PTM) in plant cells. The importance of ubiquitination in the regulation of many aspects of plant immunity has been increasingly appreciated in recent years. Most of the studies linking ubiquitination to the plant immune system, however, have been focused on the E3 ubiquitin ligases and the conventional ubiquitination that leads to the degradation of the substrate proteins by the 26S proteasome. By contrast, our knowledge about the role of unconventional ubiquitination that often serves as non‐degradative, regulatory signal remains a significant gap. We discuss, in this review, the recent advances in our understanding of ubiquitination in the modulation of plant immunity, with a particular focus on the E3 ubiquitin ligases. We approach the topic from a perspective of two broadly defined types of ubiquitination in an attempt to highlight the importance, yet current scarcity, in our knowledge about the regulation of plant immunity by unconventional ubiquitination.     

Structure and evolutionary conservation of the plant N‐end rule pathway

The N‐end rule relates the in vivo half‐life of a protein to the identity of its N‐terminal amino acid residue. While some N‐terminal residues result in metabolically stable proteins, other, so‐called destabilizing residues, lead to rapid protein turnover. The N‐end rule pathway, which mediates the recognition and degradation of proteins with N‐terminal destabilizing residues, is present in all organisms examined, including prokaryotes. This protein degradation pathway has a hierarchical organization in which some N‐terminal residues, called primary destabilizing residues, are directly recognized by specific ubiquitin ligases. Other destabilizing residues, termed secondary and tertiary destabilizing residues, require modifications before the corresponding proteins can be targeted for degradation by ubiquitin ligases. In eukaryotes, the N‐end rule pathway is a part of the ubiquitin/proteasome system and is known to play essential roles in a broad range of biological processes in fungi, animals and plants. While the structure of the N‐end rule pathway has been extensively studied in yeast and mammals, knowledge of its organization in plants is limited. Using both tobacco and Arabidopsis, we identified the complete sets destabilizing and stabilizing N‐terminal residues. We also characterized the hierarchical organization of the plant N‐end rule by identifying and determining the specificity of two distinct N‐terminal amidohydrolases (Nt‐amidases) of Arabidopsis that are essential for the destabilizing activity of the tertiary destabilizing residues Asn and Gln. Our results indicate that both the N‐end rule itself and mechanistic aspects of the N‐end rule pathway in angiosperms are very similar to those of mammals.     

Effect of polyacetylenic acids from Prunella vulgaris on various plant pathogens

Aims: This study is aiming at characterizing antifungal substances from the methanol extract of Prunella vulgaris and at investigating those substances’ antifungal and antioomycete activities against various plant pathogens. Methods and Results: Two polyacetylenic acids were isolated from P. vulgaris as active principles and identified as octadeca‐9,11,13‐triynoic acid and trans‐octadec‐13‐ene‐9,11‐diynoic acid. These two compounds inhibited the growth of Magnaporthe oryzae, Rhizoctonia solani, Phytophthora infestans, Sclerotinia sclerotiorum, Fusarium oxysporum f. sp. raphani, and Phytophthora capsici. In addition, these two compounds and the wettable powder‐type formulation of an n‐hexane fraction of P. vulgaris significantly suppressed the development of rice blast, tomato late blight, wheat leaf rust, and red pepper anthracnose. Conclusions: These data show that the extract of P. vulgaris and two polyacetylenic acids possess antifungal and antioomycete activities against a broad spectrum of tested plant pathogens. Significance and Impact of the Study: This is the first report on the occurrence of octadeca‐9,11,13‐triynoic acid and trans‐octadec‐13‐ene‐9,11‐diynoic acid in P. vulgaris and their efficacy against plant diseases. The crude extract containing the two polyacetylenic acids can be used as a natural fungicide for the control of various plant diseases.     

Sex pheromone of the alfalfa plant bug,Adelphocoris lineolatus

In northern China, due to the large‐scale adoption of transgenic Bacillus thuringiensis Berliner (Bt) cotton, the number of Adelphocoris lineolatus (Goeze) (Hemiptera: Miridae) has increased quickly, causing significant loss in cotton and alfalfa production. One of the environmentally safe strategies without use of pesticides is the application of insect pheromone for mating disruption. In our study, we aim to identify the active components in sexually mature virgin female A. lineolatus, and activity and optimal ratio of these components. By using gas chromatography–mass spectrometry (GC‐MS) and gas chromatography–electroantennographic detection (GC‐EAD), we identified three active compounds: hexyl butyrate (HB), (E)‐2‐hexenyl butyrate (E2HB), and (E)‐4‐oxo‐2‐hexenal (4‐OHE). We examined the release rate of septum and tube dispensers in a wind tunnel. In a field experiment, deletion of 4‐OHE or E2HB resulted in significant suppression of male trap catches, indicating that E2HB or 4‐OHE may be the active pheromone components. Traps baited with a blend of 4‐OHE and E2HB at 2:5, 3:4, and 4:3 caught significantly more males, suggesting that the optimal ratio of 4‐OHE and E2HB could be in the range of 1:1 to 1:2. The addition of a large amount of HB could strongly reduce the attractive activity of both virgin females and artificial lures. We also compared the attractiveness of septum lures and tube lures in field experiments. The septum lures attracted males in the first 3 days. The tube lures caught more males after 3 days and the attractive effects existed up to 5 weeks, suggesting them as a tool for long‐term monitoring and control of plant bugs.     

Roles of small RNAs in plant disease resistance

The interaction between plants and pathogens represents a dynamic competition between a robust immune system and efficient infectious strategies. Plant innate immunity is composed of complex and highly regulated molecular networks, which can be triggered by the perception of either conserved or race‐specific pathogenic molecular signatures. Small RNAs are emerging as versatile regulators of plant development, growth and response to biotic and abiotic stresses. They act in different tiers of plant immunity, including the pathogen‐associated molecular pattern‐triggered and the effector‐triggered immunity. On the other hand, pathogens have evolved effector molecules to suppress or hijack the host small RNA pathways. This leads to an arms race between plants and pathogens at the level of small RNA‐mediated defense.Here, we review recent advances in small RNA‐mediated defense responses and discuss the challenging questions in this area.     

Effects of plant quality and ant defence on herbivory rates in a neotropical ant‐plant

1. Understanding the degree to which populations and communities are limited by both bottom‐up and top‐down effects is still a major challenge for ecologists, and manipulation of plant quality, for example, can alter herbivory rates in plants. In addition, biotic defence by ants can directly influence the populations of herbivores, as demonstrated by increased rates of herbivory or increased herbivore density after ant exclusion. The aim of this study was to evaluate bottom‐up and top‐down effects on herbivory rates in a mutualistic ant‐plant. 2. In this study, the role of Azteca alfari ants as biotic defence in individuals of Cecropia pachystachya was investigated experimentally with a simultaneous manipulation of both bottom‐up (fertilisation) and top‐down (ant exclusion) factors. Four treatments were used in a fully factorial design, with 15 replicates for each treatment: (i) control plants, without manipulation; (ii) fertilised plants, ants not manipulated; (iii) unfertilised plants and excluded ants and (iv) fertilised plants and ants excluded. 3. Fertilisation increased the availability of foliar nitrogen in C. pachystachya, and herbivory rates by chewing insects were significantly higher in fertilised plants with ants excluded. 4. Herbivory, however, was more influenced by bottom‐up effects – such as the quality of the host plant – than by top‐down effects caused by ants as biotic defences, reinforcing the crucial role of leaf nutritional quality for herbivory levels experienced by plants. Conditionality in ant defence under increased nutritional quality of leaves through fertilisation might explain increased levels of herbivory in plants with higher leaf nitrogen.     

Effects of an artificial blend of host‐infested plant volatiles on plant attractiveness to parasitic wasps

For the biological control of diamondback moth (DBM) larvae in commercial greenhouses, we have previously identified a blend of volatiles that attracted Cotesia vestalis, a parasitoid of DBM larvae. Here, we tested the effects of an artificial volatile blend on the attractiveness of komatsuna plants (Japanese mustard spinach; Brassica rapa var. perviridis) to C. vestalis under greenhouse conditions. First, we showed that female C. vestalis preferred infested komatsuna plants to uninfested plants in the greenhouse. Under the same conditions, placing the artificial attractants near both infested and uninfested plants did not affect the wasps’ preference. However, when comparing infested komatsuna plants coupled with the artificial attractants with infested plants without them, significantly more female C. vestalis were attracted to the former. The possible use of artificial C. vestalis attractants for the biological control of DBM is discussed.     

Signal transduction by plant heterotrimeric G‐protein

Heterotrimeric G‐proteins are complexes that regulate important signalling pathways essential for growth and development in both plants and animals. Although plant cells are composed of the core components (Gα, Gβ and Gγ subunits) found in animal G‐proteins, the complexities of the architecture, function and signalling mechanisms of those in animals are dissimilar to those identified in some plants. Current studies on plant G‐proteins have improved knowledge of the essential physiological and agronomic properties, which when harnessed, could potentially impact global food security. Extensive studies on the molecular mechanisms underlying these properties in diverse plant species will be imperative in improving our current understanding of G‐protein signalling pathways involved in plant growth and development. The advancement of G‐protein signalling networks in distinct plant species could significantly aid in better crop development. This review summarizes current progress, novel discoveries and future prospects for this area in potential crop improvement.     

Relative climatic,edaphic and management controls of plant functional trait signatures

Objective: To identify the relative roles of climatic, edaphic and management factors in controlling the weighted mean traits of vegetation. Location: Eleven sites in Europe and one in Israel undergoing transitions in land use. Material and Methods: Standardised methods were used to collect information on species traits and attributes from plots covering a range of land uses at each site. This was combined with abundance data to create a plot × trait matrix. Variance partitioning was used to identify the relative roles of climate, soil and management on the weighted and unweighted mean traits of the vegetation in the full data set, and the data set divided into vegetative traits (including life‐form, clonality, defence and a range of leaf traits) and traits linked to regeneration via seeds (including seed mass, dispersal and pollination mechanism). Results: Variance partitioning of the full data set showed that climate (18.7%), explained more variance in the weighted mean traits of the vegetation than climate and soil together (9.2), soil (6.9) and management (6.1). There was a similar distribution of variance explained for both vegetative and regeneration via seed traits, although more variance was explained for the latter. This restricted set of climatic, edaphic and management variables could explain 45‐50% of the variance in the weighted mean traits of the vegetation between plots. There were only small differences between analyses of the weighted and unweighted data. Conclusions: Despite large variations in climate and soils between sites, there was still a separate and recognisable impact of management on the mean weighted traits of the vegetation. There was also a degree of shared variation between the three groups of factors, indicating that the response of plant traits to one group of factors may not be predictable because they may be modulated by their response to other groups.     

Translational genomics for plant breeding with the genome sequence explosion

The use of next‐generation sequencers and advanced genotyping technologies has propelled the field of plant genomics in model crops and plants and enhanced the discovery of hidden bridges between genotypes and phenotypes. The newly generated reference sequences of unstudied minor plants can be annotated by the knowledge of model plants via translational genomics approaches. Here, we reviewed the strategies of translational genomics and suggested perspectives on the current databases of genomic resources and the database structures of translated information on the new genome. As a draft picture of phenotypic annotation, translational genomics on newly sequenced plants will provide valuable assistance for breeders and researchers who are interested in genetic studies.     

Bacterial and plant natriuretic peptides improve plant defence responses against pathogens

Plant natriuretic peptides (PNPs) have been implicated in the regulation of ions and water homeostasis, and their participation in the plant immune response has also been proposed. Xanthomonas citri ssp. citri contains a gene encoding a PNP‐like protein (XacPNP) which has no homologues in other bacteria. XacPNP mimics its Arabidopsis thaliana homologue AtPNP‐A by modifying host responses to create favourable conditions for pathogen survival. However, the ability of XacPNP to induce plant defence responses has not been investigated. In order to study further the role of XacPNP in vivo, A. thaliana lines over‐expressing XacPNP, lines over‐expressing AtPNP‐A and AtPNP‐A‐deficient plants were generated. Plants over‐expressing XacPNP or AtPNP‐A showed larger stomatal aperture and were more resistant to saline or oxidative stress than were PNP‐deficient lines. In order to study further the role of PNP in biotic stress responses, A. thaliana leaves were infiltrated with pure recombinant XacPNP, and showed enhanced expression of genes related to the defence response and a higher resistance to pathogen infections. Moreover, AtPNP‐A expression increased in A. thaliana on Pseudomonas syringae pv. tomato (Pst) infection. This evidence led us to analyse the responses of the transgenic plants to pathogens. Plants over‐expressing XacPNP or AtPNP‐A were more resistant to Pst infection than control plants, whereas PNP‐deficient plants were more susceptible and showed a stronger hypersensitive response when challenged with non‐host bacteria. Therefore, XacPNP, acquired by horizontal gene transfer, is able to mimic PNP functions, even with an increase in plant defence responses.     

The in vivo synthesis of plant sesquiterpenes by Escherichia coli.

Three plant genes encoding (+)-delta-cadinene, 5-epi-aristolochene, and vetispiradiene cyclases were expressed in Escherichia coli to evaluate the potential of this bacterium to synthesize sesquiterpenes in vivo. Various growth temperatures, carbon sources, and host strains were examined to optimize terpene production. The highest levels of sesquiterpene production occurred when the enzymes were expressed in strain DH5alpha from the trc promoter (Ptrc) of the high-copy plasmidpTrc99A in M9 medium supplemented with 0.2% (v/v) glycerol at 30 degrees C for 5-epi-aristolochene and vetispiradiene and 37 degrees C for (+)-delta-cadinene. The highest concentrations of sesquiterpenes observed were 10.3 microg of (+)-delta-cadinene, 0.24 microg of 5-epi-aristolochene (measured as (+)-delta-cadinene equivalents), and 6.4 microg of vetispiradiene (measured as (+)-delta-cadinene equivalents) per liter of culture. These sesquiterpene production levels are >500-fold lower than carotenoid production, both of which are synthesized from endogenous trans-farnesyl diphosphate (FDP) in E. coli. Based on these results, we conclude that the limiting factor for sesquiterpene synthesis in E. coli is the poor expression of the cyclase enzyme and not supply of the FDP precursor.     

Interaction of plant polysomes with the actin cytoskeleton

Protein composition and functional activity of various polysome subpopulations isolated from Vicia faba L. leaves and Triticum aestivum L. and Hordeum vulgare L. seedlings were studied. Membrane- and cytoskeleton-bound polysomes were more active in the wheat germ cell-free translational system than free polysomes. Several non-ribosomal proteins were detected in the polysome preparations by gel electrophoresis and Western blot analysis: (1) a canonical actin of mol wt 42 kDa; (2) a 40 kDa protein, demonstrating affinity for ribosomes, sharing some determinants with actin, and present predominantly in the subpopulations of bound polysomes; and (3) an acidic ribosome-associated p40 evenly distributed between free and bound polysomes. The possibility of involvement of these proteins in interactions between polysomes and the actin cytoskeleton is discussed.     

Differential responses of soil bacteria,fungi, archaea and protists to plant species richness and plant functional group identity

Plants are known to influence belowground microbial community structure along their roots, but the impacts of plant species richness and plant functional group (FG) identity on microbial communities in the bulk soil are still not well understood. Here, we used 454‐pyrosequencing to analyse the soil microbial community composition in a long‐term biodiversity experiment at Jena, Germany. We examined responses of bacteria, fungi, archaea, and protists to plant species richness (communities varying from 1 to 60 sown species) and plant FG identity (grasses, legumes, small herbs, tall herbs) in bulk soil. We hypothesized that plant species richness and FG identity would alter microbial community composition and have a positive impact on microbial species richness. Plant species richness had a marginal positive effect on the richness of fungi, but we observed no such effect on bacteria, archaea and protists. Plant species richness also did not have a large impact on microbial community composition. Rather, abiotic soil properties partially explained the community composition of bacteria, fungi, arbuscular mycorrhizal fungi (AMF), archaea and protists. Plant FG richness did not impact microbial community composition; however, plant FG identity was more effective. Bacterial richness was highest in legume plots and lowest in small herb plots, and AMF and archaeal community composition in legume plant communities was distinct from that in communities composed of other plant FGs. We conclude that soil microbial community composition in bulk soil is influenced more by changes in plant FG composition and abiotic soil properties, than by changes in plant species richness per se.     

Emerging role of roots in plant responses to aboveground insect herbivory

Plants have evolved complex biochemical mechanisms to counter threats from insect herbivory. Recent research has revealed an important role of roots in plant responses to above ground herbivory (AGH). The involvement of roots is integral to plant resistance and tolerance mechanisms. Roots not only play an active role in plant defenses by acting as sites for biosynthesis of various toxins and but also contribute to tolerance by storing photoassimilates to enable future regrowth. The interaction of roots with beneficial soil‐borne microorganisms also influences the outcome of the interaction between plant and insect herbivores. Shoot‐to‐root communication signals are critical for plant response to AGH. A better understanding of the role of roots in plant response to AGH is essential in order to develop a comprehensive picture of plant‐insect interactions. Here, we summarize the current status of research on the role of roots in plant response to AGH and also discuss possible signals involved in shoot‐to‐root communication.     

Integration of two herbivore‐induced plant volatiles results in synergistic effects on plant defence and resistance

Plants can use induced volatiles to detect herbivore‐ and pathogen‐attacked neighbors and prime their defenses. Several individual volatile priming cues have been identified, but whether plants are able to integrate multiple cues from stress‐related volatile blends remains poorly understood. Here, we investigated how maize plants respond to two herbivore‐induced volatile priming cues with complementary information content, the green leaf volatile (Z)‐3‐hexenyl acetate (HAC) and the aromatic volatile indole. In the absence of herbivory, HAC directly induced defence gene expression, whereas indole had no effect. Upon induction by simulated herbivory, both volatiles increased jasmonate signalling, defence gene expression, and defensive secondary metabolite production and increased plant resistance. Plant resistance to caterpillars was more strongly induced in dual volatile‐exposed plants than plants exposed to single volatiles.. Induced defence levels in dual volatile‐exposed plants were significantly higher than predicted from the added effects of the individual volatiles, with the exception of induced plant volatile production, which showed no increase upon dual‐exposure relative to single exposure. Thus, plants can integrate different volatile cues into strong and specific responses that promote herbivore defence induction and resistance. Integrating multiple volatiles may be beneficial, as volatile blends are more reliable indicators of future stress than single cues.