Induced Plant Defense Research Articles

A chitosan-coated lentinan-loaded calcium alginate hydrogel induces broad-spectrum resistance to plant viruses by activating Nicotiana benthamiana calmodulin-like (CML) protein 3.

Control of plant virus diseases largely depends on the induced plant defence achieved by the external application of synthetic chemical inducers with the ability to modify defence-signalling pathways. However, most of the molecular mechanisms underlying these chemical inducers remain unknown. Here, we developed a chitosan-coated lentinan-loaded hydrogel and discovered how it protects plants from different virus infections. The hydrogel was synthesized by coating chitosan on the surface of the calcium alginate-lentinan (LNT) hydrogel (SL-gel) to form a CSL-gel. CSL-gels exhibit the capacity to prolong the stable release of lentinan and promote Ca2+ release. Application of CSL-gels on the root of plants induces broad-spectrum resistance against plant viruses (TMV, TRV, PVX and TuMV). RNA-seq analysis identified that Nicotiana benthamiana calmodulin-like protein gene 3 (NbCML3) is upregulated by the sustained release of Ca2+ from the CSL-gel, and silencing and overexpression of NbCML alter the susceptibility and resistance of tobacco to TMV. Our findings provide evidence that this novel and synthetic CSL-gel strongly inhibits the infection of plant viruses by the sustainable release of LNT and Ca2+ . This study uncovers a novel mode of action by which CSL-gels trigger NbCML3 expression through the stable and sustained release of Ca2+ .

Bacterial volatiles as PGPRs: Inducing plant defense mechanisms during stress periods

The increase in population and food demand has led to the excessive utilization of chemical fertilizers. These fertilizers, in turn, affected the soil properties and decreased the fertility of the soil. The side effects of these chemicals have forced scientists to focus on plant growth-promoting rhizobacteria (PGPR) for sustainable farming. Some microbes and plants have a mutual relationship, and they use to help each other in various ways Rhizobacteria that promote plant growth enhance health of the plant and growth by increasing nutrient accessibility and assisting plants during abiotic and biotic stress. By direct or indirect mechanisms, the bacterial releases of volatile organic compounds (VOCs) are crucial in boosting plant development and disease resistance. The release of a range of low-molecular weight hydrocarbons, including the growth-promoting volatile organic compounds, 3‑hydroxy-2-butanone (acetone), 2,3-butanediol, 2-pentylfuran, or dimethylhexadecylamine, was found by chemical analysis of bacterial volatile emissions. The plants provide nutrients and shelter to the microbes. Therefore, PGPRs play a vital role in sustainable agriculture In this review, the mechanism of action of VOCs for growth promotion and how they trigger the plant defense mechanism during biotic and abiotic stress will be addressed.This review will addressthe mechanism of action of VOCs for growth promotion and how they trigger the plant defense mechanism during biotic and abiotic stress.

Advances in understanding the mechanism of resistance to anthracnose and induced defence response in tea plants.

The tea plant (Camellia sinensis) is susceptible to anthracnose disease that causes considerable crop loss and affects the yield and quality of tea. Multiple Colletotrichum spp. are the causative agents of this disease, which spreads quickly in warm and humid climates. During plant-pathogen interactions, resistant cultivars defend themselves against the hemibiotrophic pathogen by activating defence signalling pathways, whereas the pathogen suppresses plant defences in susceptible varieties. Various fungicides have been used to control this disease on susceptible plants, but these fungicide residues are dangerous to human health and cause fungicide resistance in pathogens. The problem-solving approaches to date are the development of resistant cultivars and ecofriendly biocontrol strategies to achieve sustainable tea cultivation and production. Understanding the infection stages of Colletotrichum, tea plant resistance mechanisms, and induced plant defence against Colletotrichum is essential to support sustainable disease management practices in the field. This review therefore summarizes the current knowledge of the identified causative agent of tea plant anthracnose, the infection strategies and pathogenicity of C. gloeosporioides, anthracnose disease resistance mechanisms, and the caffeine-induced defence response against Colletotrichum infection. The information reported in this review will advance our understanding of host-pathogen interactions and eventually help us to develop new disease control strategies.

Aphid infestations reduce monarch butterfly colonization, herbivory, and growth on ornamental milkweed.

Anthropogenic disturbance is driving global biodiversity loss, including the monarch butterfly (Danaus plexippus), a dietary specialist of milkweed. In response, ornamental milkweed plantings are increasingly common in urbanized landscapes, and recent evidence indicates they have conservation value for monarch butterflies. Unfortunately, sap-feeding insect herbivores, including the oleander aphid (Aphis nerii), frequently reach high densities on plants in nursery settings and urbanized landscapes. Aphid-infested milkweed may inhibit monarch conservation efforts by reducing host plant quality and inducing plant defenses. To test this, we evaluated the effects of oleander aphid infestation on monarch oviposition, larval performance, and plant traits using tropical milkweed (Asclepias curassavica), the most common commercially available milkweed species in the southern U.S. We quantified monarch oviposition preference, larval herbivory, larval weight, and plant characteristics on aphid-free and aphid-infested milkweed. Monarch butterflies deposited three times more eggs on aphid-free versus aphid-infested milkweed. Similarly, larvae fed aphid-free milkweed consumed and weighed twice as much as larvae fed aphid-infested milkweed. Aphid-free milkweed had higher total dry leaf biomass and nitrogen content than aphid-infested milkweed. Our results indicate that oleander aphid infestations can have indirect negative impacts on urban monarch conservation efforts and highlight the need for effective Lepidoptera-friendly integrated pest management tactics for ornamental plants.

Oviposition by Plagiodera versicolora on Salix matsudana cv. 'Zhuliu' alters the leaf transcriptome and impairs larval performance.

Insect egg deposition can induce plant defenses against their larvae. Previous studies have primarily focused on herbaceous plant defenses; however, little is known about how the Salicaceae respond to insect egg deposition and defend themselves against herbivores. By combining plant defense gene studies and bioassays, we investigated the effect of the coleoptera Plagiodera versicolora egg deposition on willow (Salix matsudana cv. 'Zhuliu') and examined the interactions at the plant resistance and transcriptome levels. RNA-seq data were utilized to analyze changes in the leaf transcriptome with and without oviposition, and also the changes in the leaf transcriptome of feeding-damaged leaves with and without prior oviposition. P. versicolora oviposition on willow leaves resulted in altered expression levels of transcripts associated with plant stress and metabolic responses. Compared with leaves with no oviposition, leaves with egg deposition showed a slight increase in phenylpropanoid biosynthesis and phytohormone signaling genes after larval feeding. The RNA-seq analysis revealed alterations in willow transcripts in response to leaf beetle infestations. Bioassays indicated that oviposition by P. versicolora on willows reduced subsequent larvae performance, suggesting that prior oviposition by P. versicolora could increase willows' resistance to larvae. This study advances our knowledge of how oviposition by coleoptera insects induces changes in the resistance of leaves to herbivory in the Salicaceae family.

Melatonin and strigolactone mitigate chromium toxicity through modulation of ascorbate-glutathione pathway and gene expression in tomato

Chromium (Cr) is considered one of the most hazardous metal contaminant reducing crop production and putting human health at risk. Phytohormones are known to regulate chromium stress, however, the function of melatonin and strigolactones in Chromium stress tolerance in tomato is rarely investigated. Here we investigated the potential role of melatonin (ML) and strigolactone (SL) on mitigating Chromium toxicity in tomato. With exposure to 300 μM Cr stress a remarkable decline in growth (63.01%), biomass yield (50.25)%, Pigment content (24.32%), photosynthesis, gas exchange and Physico-biochemical attributes of tomato was observed. Cr treatment also resulted in oxidative stress closely associated with higher H2O2 generation (215.66%), Lipid peroxidation (50.29%), electrolyte leakage (440.01%) and accumulation of osmolytes like proline and glycine betine. Moreover, Cr toxicity up-regulated the transcriptional expression profiles of antioxidant, stress related and metal transporter genes and down-regulated the genes related to photosynthesis. The application of ML and SL alleviated the Cr induced phytotoxic effects on photosynthetic pigments, gas exchange parameters and restored growth of tomato plants. ML and SL supplementation induced plant defense system via enhanced regulation of antioxidant enzymes, ascorbate and glutathione pool and transcriptional regulation of several genes. The coordinated regulation of antioxidant and glyoxalase systems expressively suppressed the oxidative stress. Hence, ML and SL application might be considered as an effective approach for minimizing Cr uptake and its detrimental effects in tomato plants grown in contaminated soils. The study may also provide new insights into the role of transcriptional regulation in the protection against heavy metal toxicity.

How does the pattern of root metabolites regulating beneficial microorganisms change with different grazing pressures?

Grazing disturbance can change the structure of plant rhizosphere microbial communities and thereby alter the feedback to promote plant growth or induce plant defenses. However, little is known about how such changes occur and vary under different grazing pressures or the roles of root metabolites in altering the composition of rhizosphere microbial communities. In this study, the effects of different grazing pressures on the composition of microbial communities were investigated, and the mechanisms by which different grazing pressures changed rhizosphere microbiomes were explored with metabolomics. Grazing changed composition, functions, and co-expression networks of microbial communities. Under light grazing (LG), some saprophytic fungi, such as Lentinus sp., Ramichloridium sp., Ascobolus sp. and Hyphoderma sp., were significantly enriched, whereas under heavy grazing (HG), potentially beneficial rhizobacteria, such as Stenotrophomonas sp., Microbacterium sp., and Lysobacter sp., were significantly enriched. The beneficial mycorrhizal fungus Schizothecium sp. was significantly enriched in both LG and HG. Moreover, all enriched beneficial microorganisms were positively correlated with root metabolites, including amino acids (AAs), short-chain organic acids (SCOAs), and alkaloids. This suggests that these significantly enriched rhizosphere microbial changes may be caused by these differential root metabolites. Under LG, it is inferred that root metabolites, especially AAs such as L-Histidine, may regulate specific saprophytic fungi to participate in material transformations and the energy cycle and promote plant growth. Furthermore, to help alleviate the stress of HG and improve plant defenses, it is inferred that the root system actively regulates the synthesis of these root metabolites such as AAs, SCOAs, and alkaloids under grazing interference, and then secretes them to promote the growth of some specific plant growth-promoting rhizobacteria and fungi. To summarize, grasses can regulate beneficial microorganisms by changing root metabolites composition, and the response strategies vary under different grazing pressure in typical grassland ecosystems.

Fungal Endophytes: An Alternative Biocontrol Agent against Phytopathogenic Fungi

There has been renewed interest in the application of endophytic fungi to control phytopathogenic fungi, which cause significant damage to crop health, ultimately leading to losses in agricultural productivity. Endophytic fungi inhibit pathogens via different modes of action—mycoparasitism, competition (for nutrients and ecological niches), antibiosis, and induction of plant defense—thus demonstrating the ability to control a wide range of phytopathogenic fungi in different growth phases and habitats. However, many studies have been conducted under laboratory conditions, and there is a huge lack of studies in which real field testing was performed. Aspergillus, Clonostachys, Coniothyrium, Trichoderma, and Verticillium have been proven to be the most effective fungal biocontrol agents. Trichoderma is regarded as the most promising group in commercial formulations. In this study, we attempted to emphasize the significance of fungal endophytes in controlling phytopathogenic fungi, while reporting recent advances in endophytic biology and application.

Rhizoctonia solani AG1 IA extracellular polysaccharides: Structural characterization and induced resistance to rice sheath blight

Sheath blight, caused by Rhizoctonia solani (R. solani), is one of the most serious diseases of rice. Extracellular polysaccharides (EPS) are complex polysaccharides secreted by microbes that have a pivotal role in the plant-microbe interaction. At present, many studies have been carried out on R. solani, but it is not very clear whether the EPS is secreted by R. solani exists. Therefore, we isolated and extracted the EPS from R. solani, two kinds of EPS (EW-I and ES-I) were obtained by DEAE-cellulose 52 and Sephacryl S-300HR column further purification, and their structures were characterized by FT-IR, GC-MS, and NMR analysis. The results showed that EW-I and ES-I had similar monosaccharide composition but different molar ratio, they were composed of fucose, arabinose, galactose, glucose, and mannose with a ratio of 7.49: 27.72: 2.98: 6.66: 55.15 and 3.81: 12.98: 6.15: 10.83: 66.23, and their backbone may be composed of →2)-α-Manp-(1→ residues, beside ES-I was highly branched compared to EW-I. The exogenous application of EW-I and ES-I had no effect on the growth of R. solani AG1 IA itself, but their pretreatment of rice induced plant defense through activation of the salicylic acid pathway, resulting in enhanced resistance to sheath blight.

Ubiquitin E3 ligase activity of Ralstonia solanacearum effector RipAW is not essential for induction of plant defense in Nicotiana benthamiana.

As one of the most destructive bacterial phytopathogens, Ralstonia solanacearum causes substantial annual yield losses of many important crops. Deciphering the functional mechanisms of type III effectors, the crucial factors mediating R. solanacearum-plant interactions, will provide a valuable basis for protecting crop plants from R. solanacearum. Recently, the NEL (novel E3 ligase) effector RipAW was found to induce cell death on Nicotiana benthamiana in a E3 ligase activity-dependent manner. Here, we further deciphered the role of the E3 ligase activity in RipAW-triggered plant immunity. We found that RipAWC177A, the E3 ligase mutant of RipAW, could not induce cell death but retained the ability of triggering plant immunity in N. benthamiana, indicating that the E3 ligase activity is not essential for RipAW-triggered immunity. By generating truncated mutants of RipAW, we further showed that the N-terminus, NEL domain and C-terminus are all required but not sufficient for RipAW-induced cell death. Furthermore, all truncated mutants of RipAW triggered ETI immune responses in N. benthamiana, confirming that the E3 ligase activity is not essential for RipAW-triggered plant immunity. Finally, we demonstrated that RipAW- and RipAWC177A-triggered immunity in N. benthamiana requires SGT1 (suppressor of G2 allele of skp1), but not EDS1 (enhanced disease susceptibility), NRG1 (N requirement gene 1), NRC (NLR required for cell death) proteins or SA (salicylic acid) pathway. Our findings provide a typical case in which the effector-induced cell death can be uncoupled with immune responses, shedding new light on effector-triggered plant immunity. Our data also provide clues for further in-depth study of mechanism underlying RipAW-induced plant immunity.

Variation in the Outcome of Plant-Mediated Pathogen Interactions in Potato: Effects of Initial Infections on Conspecific vs. Heterospecific Subsequent Infections

Plants are often attacked sequentially by multiple enemies. Pathogen sequential co-infections can lead to indirect interactions mediated by plant induced responses whose outcome is contingent on differences in the magnitude and type of plant induced defences elicited by different species or guilds. To date, however, most studies have tested unidirectional effects of one pathogen on another, not discerning between conspecific vs. heterospecific infections, and often not measuring plant induced responses underlying such outcomes. To address this, we conducted a greenhouse experiment testing for the impact of initial infection by two leaf pathogens (Alternaria solani and Phytophthora infestans) on subsequent infection by each of these pathogens on potato (Solanum tuberosum) plants, and also measured induced plant defences (phenolic compounds) to inform on interaction outcomes. We found contrasting results depending on the identity of the initially infecting pathogen. Specifically, initial infection by A. solani drove induced resistance (lower necrosis) by subsequently infecting A. solani (conspecific induced resistance) but had no effect on subsequent infection by P. infestans. In contrast, initial infection by P. infestans drove induced resistance to subsequent infection by both conspecifics and A. solani. Patterns of plant induced defences correlated with (and potentially explained) induced resistance to conspecific but not heterospecific (e.g., in the case of P. infestans) subsequent infection. Overall, these results further our understanding of plant-mediated pathogen interactions by showing that plant-mediated interactions between pathogen species can be asymmetrical and in some cases not reciprocal, that pathogen species can vary in the importance of conspecific vs. heterospecific effects, and shed mechanistic insight into the role of plant induced responses driving such interactions.

The Fall Armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae), Influences Nilaparvata lugens Population Growth Directly, by Preying on Its Eggs, and Indirectly, by Inducing Defenses in Rice.

The fall armyworm (FAW), Spodoptera frugiperda, has become one of the most important pests on corn in China since it invaded in 2019. Although FAW has not been reported to cause widespread damage to rice plants in China, it has been sporadically found feeding in the field. If FAW infests rice in China, the fitness of other insect pests on rice may be influenced. However, how FAW and other insect pests on rice interact remains unknown. In this study, we found that the infestation of FAW larvae on rice plants prolonged the developmental duration of the brown planthopper (BPH, Nilaparvata lugens (Stål)) eggs and plants damaged by gravid BPH females did not induce defenses that influenced the growth of FAW larvae. Moreover, co-infestation by FAW larvae on rice plants did not influence the attractiveness of volatiles emitted from BPH-infested plants to Anagrus nilaparvatae, an egg parasitoid of rice planthoppers. FAW larvae were able to prey on BPH eggs laid on rice plants and grew faster compared to those larvae that lacked available eggs. Studies revealed that the delay in the development of BPH eggs on FAW-infested plants was probably related to the increase in levels of jasmonoyl-isoleucine, abscisic acid and the defensive compounds in the rice leaf sheaths on which BPH eggs were laid. These findings indicate that, if FAW invades rice plants in China, the population density of BPH may be decreased by intraguild predation and induced plant defenses, whereas the population density of FAW may be increased.

Experimental Growth Conditions affect Direct and Indirect Defences in two Cotton Species

Cotton has been used as a model plant to study direct and indirect plant defence against herbivorous insects. However, the plant growing conditions could have an important effect on the outcome of such plant defence studies. We examined how common experimental growth conditions influence constitutive and inducible defences in two species of cotton, Gossypium hirsutum and G. herbaceum. We induced plants by applying caterpillar regurgitant to mechanical wounds to compare the induction levels between plants of both species grown in greenhouse or phytotron conditions. For this we measured defence metabolites (gossypol and heliocides) and performance of Spodoptera frugiperda caterpillars on different leaves, the emission of plant volatiles, and their attractiveness to parasitic wasps. Induction increased the levels of defence metabolites, which in turn decreased the performance of S. frugiperda larvae. Constitutive and induced defence levels were the highest in plants grown in the phytotron (compared to greenhouse plants), G. hirsutum and young leaves. Defence induction was more pronounced in plants grown in the phytotron and in young leaves. Also, the differences between growing conditions were more evident for metabolites in the youngest leaves, indicating an interaction with plant ontogeny. The composition of emitted volatiles was different between plants from the two growth conditions, with greenhouse-grown plants showing more variation than phytotron-grown plants. Also, G. hirsutum released higher amounts of volatiles and attracted more parasitic wasps than G. herbaceum. Overall, these results highlight the importance of experimental abiotic factors in plant defence induction and ontogeny of defences. We therefore suggest careful consideration in selecting the appropriate experimental growing conditions for studies on plant defences.

Pectate Lyase from Fusarium sacchari Induces Plant Immune Responses and Contributes to Virulence.

Fusarium sacchari is one of the primary pathogens causing Pokkah Boeng disease (PBD) in sugarcane in China. Pectate lyases (PL), which play a critical role in pectin degradation and fungal virulence, have been extensively studied in major bacterial and fungal pathogens of a wide range of plant species. However, only a few PLs have been functionally investigated. In this study, we analyzed the function of the pectate lyase gene, FsPL, from F. sacchari. FsPL is a key virulence factor of F. sacchari and can induce plant cell death. FsPL also triggers the pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) response in Nicotiana benthamiana, as reflected by increases in reactive oxygen species (ROS) production, electrolyte leakage, and callose accumulation, as well as the upregulation of defense response genes. In addition, our study also found that the signal peptide of FsPL was necessary for induced cell death and PTI responses. Virus-induced gene silencing showed that FsPL-induced cell death in Nicotiana benthamiana was mediated by leucine-rich repeat (LRR) receptor-like kinases BAK1 and SOBIR1. Thus, FsPL may not only be a critical virulence factor for F. sacchari but may also induce plant defense responses. These findings provide new insights into the functions of pectate lyase in host-pathogen interactions. IMPORTANCE Pokkah Boeng disease (PBD) is one of the main diseases affecting sugarcane in China, seriously damaging sugarcane production and economic development. Therefore, it is important to clarify the pathogenic mechanisms of this disease and to provide a theoretical basis for the breeding of PBD-resistant sugarcane strains. The present study aimed to analyze the function of FsPL, a recently identified pectate lyase gene from F. sacchari. FsPL is a key virulence factor of F. sacchari that induces plant cell death. Our results provide new insights into the function of pectate lyase in host-pathogen interactions.

Naïve Pine Terpene Response to the Mountain Pine Beetle (Dendroctonus ponderosae) through the Seasons.

Insect herbivores must contend with constitutive and induced plant defenses. The mountain pine beetle, Dendroctonus ponderosae Hopkins (Coleoptera: Curculionidae, Scolytinae) has expanded its range east of the Rocky Mountains into the western boreal forest and is encountering evolutionarily naïve lodgepole pines (Pinus contorta) and jack pines (Pinus banksiana). Pinus contorta and P. banksiana in the expanded range have different constitutive and induced defenses in response to wounding and inoculation with fungal associates of D. ponderosae. In the historic range, previous studies have examined phloem terpene content prior to and just after D. ponderosae mass attack, but the terpene profile of attacked trees post-overwintering is unknown. We examined the response of mature P. contorta and P. banksiana trees to experimentally-applied mass attack by D. ponderosae and quantified phloem terpenes at three time points, pre-attack, post-attack (same season), and the following spring, post-overwintering. Phloem content of total terpenes as well as many individual terpenes increased after D. ponderosae attack but were only significantly higher than pre-attack levels at the post-overwintering time point in both P. contorta and P. banksiana. The absence of a significant increase in phloem terpenes in the month following attack in naïve pines is a potential cause for increased D. ponderosae offspring production reported in naïve P. contorta. Beetle attack density did not influence the phloem terpene profiles of either species and there was no significant interaction between attack density and sampling time on terpene content. High phloem terpenes in trees that are attacked at low densities could prime these trees for defense against attacks in the following season but it could also make these trees more apparent to early-foraging beetles and facilitate efficient mass attack at low D. ponderosae population densities in the expanded range.

Bactericidal and plant defense elicitation activities of Eucalyptus oil decrease the severity of infections by Xylella fastidiosa on almond plants.

The activity of Eucalyptus essential oil against eleven strains pertaining to six species of plant pathogenic bacteria was studied using growth inhibition and contact assays. All strains were susceptible to the formulation EGL2, and Xylella fastidiosa subspecies and Xanthomonas fragariae were the most sensitive. The bactericidal effect was strong causing 4.5 to 6.0 log reductions in survival in 30min at concentrations in the range of 0.75 to 15.0 μl/ml depending on the bacteria tested. Transmission electron microscopy of the formulation EGL2 against the three X. fastidiosa subspecies studied allowed the observation of a strong lytic effect on bacterial cells. In addition, the preventive spray application of EGL2 to potted pear plants subsequently inoculated with Erwinia amylovora significantly decreased the severity of infections. Almond plants treated by endotherapy or soil drenching, and then inoculated with X. fastidiosa showed a significant decrease in disease severity as well as in the levels of the pathogen, depending on the strategy used (endotherapy/soil drenching, preventive/curative). The treatment by endotherapy in almond plants induced the expression of several genes involved in plant defense. It was concluded that the reduction of infections by the Eucalyptus oil treatments was due to the combination of its bactericidal and plant defense induction activities.

Disentangling the resistant mechanism of Fusarium wilt TR4 interactions with different cultivars and its elicitor application.

Fusarium wilt of banana, especially Tropical Race 4 (TR4) is a major factor restricting banana production. Developing a resistant cultivar and inducing plant defenses by elicitor application are currently two of the best options to control this disease. Isotianil is a monocarboxylic acid amide that has been used as a fungicide to control rice blast and could potentially induce systemic acquired resistance in plants. To determine the control effect of elicitor isotianil on TR4 in different resistant cultivars, a greenhouse pot experiment was conducted and its results showed that isotianil could significantly alleviate the symptoms of TR4, provide enhanced disease control on the cultivars 'Baxi' and 'Yunjiao No.1' with control effect 50.14% and 56.14%, respectively. We compared the infection processes in 'Baxi' (susceptible cultivars) and 'Yunjiao No.1' (resistant cultivars) two cultivars inoculated with pathogen TR4. The results showed that TR4 hyphae could rapidly penetrate the cortex into the root vascular bundle for colonization, and the colonization capacity in 'Baxi' was significantly higher than that in 'Yunjiao No.1'. The accumulation of a large number of starch grains was observed in corms cells, and further analysis showed that the starch content in 'Yunjiao No. 1' as resistant cultivar was significantly higher than that in 'Baxi' as susceptible cultivar, and isotianil application could significantly increase the starch content in 'Baxi'. Besides, a mass of tyloses were observed in the roots and corms and these tyloses increased after application with isotianil. Furthermore, the total starch and tyloses contents and the control effect in the corms of 'Yunjiao No.1' was higher than that in the 'Baxi'. Moreover, the expression levels of key genes for plant resistance induction and starch synthesis were analyzed, and the results suggested that these genes were significantly upregulated at different time points after the application of isotianil. These results suggest that there are significant differences between cultivars in response to TR4 invasion and plant reactions with respect to starch accumulation, tyloses formation and the expression of plant resistance induction and starch synthesis related genes. Results also indicate that isotianil application may contribute to disease control by inducing host plant defense against TR4 infection and could be potentially used together with resistant cultivar as integrated approach to manage this destructive disease. Further research under field conditions should be included in the next phases of study.

Postharvest Application of Acibenzolar-S-Methyl Activates Salicylic Acid Pathway Genes in Kiwifruit Vines.

The plant defence inducer Actigard® (acibenzolar-S-methyl [ASM]) is applied before flowering and after fruit harvest to control bacterial canker in kiwifruit caused by Pseudomonas syringae pv. actinidiae. Pre-flowering application of ASM is known to upregulate defence gene expression; however, the effect of postharvest ASM on defence gene expression in the vine is unknown. In this study, the expression of eight "defence marker" genes was measured in the leaves of Actinidia chinensis var. chinensis, "Zesy002," and Actinidia chinensis var. deliciosa, "Hayward," vines after postharvest treatment with ASM and/or copper. There were two orchards per cultivar with harvest dates approximately three weeks apart for investigating potential changes in responsiveness to ASM during the harvest period. In all trials, postharvest ASM induced the expression of salicylic-acid-pathway defence genes PR1, PR2, PR5, BAD, DMR6, NIMIN2, and WRKY70. Gene upregulation was the greatest at 1 day and 7 days after treatment and declined to the control level after 3 weeks. In "Zesy002", the ASM-induced response was greater at the early harvest site than at the late harvest site. This decline was concomitant with leaf yellowing and a reduction in RNA yield. Effects of postharvest ASM on gene expression did not persist into the following spring, nor were vines conditioned to respond more strongly to pre-flowering ASM application.

Diketopiperazine cyclo(-l-Leu-l-Phe) with plant elicitation activity and anti-oomycete activity against Plasmopara viticola.

The aim of this study was to contribute to the reduction of synthetic chemical fungicide application in viticulture by using cyclo(-l-Leu-l-Phe) (cLF) produced by Bacillus subtilis KS1, a candidate for biological control agent. cLF is a diketopiperazine and belongs to the family of 2,5-diketopiperazines. KS1 secreted micromolar levels of cLF into culture medium. Micromolar concentrations of cLF structure-dependently decreased by ∼90% both disease severity and lesion density of downy mildew in grapevine cultivated in a growth chamber. Microscopic observation demonstrated that cLF inhibited Plasmopara viticola haustorium formation by 80% but not zoospore germination on leas disks. Interestingly, millimolar concentrations of cLF induced plant defense response, such as expression of genes encoding chitinase and β-1,3-glucanase, in grapevine leaves through the salicylic acid and jasmonate signaling pathways. We demonstrated that cLF was a weapon against P. viticola infection. Further evaluation of cLF in field trials is required to uncover its inherent characteristics.

Identification of dominant taxa of sooty moulds and their impact on the leaf microbiome.

Sooty moulds are a widespread group of saprophytic ascomycetes that obtain nutrients from honeydew excreted by sap-feeding insects and coat plant tissue with mycelia. Research on sooty moulds has focused on fungal morphology and phylogeny-based taxonomy, but little research has been conducted on the community structure. In this study, the PacBio sequencing platform was used to systematically analyse the fungal and bacterial diversity of the sooty mould community on camphor trees at two sampling sites. Six dominant sooty mould genera were identified, of which three genera of Dothideomycetes were enriched only in diseased samples, while three genera of Eurotiomycetes were present in both healthy and diseased samples. Bacterial diversity and co-occurrence network analysis indicated that the sooty moulds had an effect on the leaf surface bacterial communities but not on the endophyte communities. There was a close correlation between the six dominant pathogenic groups and bacteria in the soot layer. Transcriptomic data from Cinnamomum camphora samples showed that the sooty moulds that did not penetrate plant cells not only affected plant photosynthesis but also induced plant defence responses. This study systematically studied the microbial community of sooty moulds, indicating the close relationship between sooty moulds and the bacterial communities.