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1.
Hortic Res ; 10(10): uhad178, 2023 Oct.
Article in English | MEDLINE | ID: mdl-37868619

ABSTRACT

Protease inhibitors promote herbivore resistance in diverse plant species. Although many inducible protease inhibitors have been identified, there are limited reports available on the biological relevance and molecular basis of constitutive protease inhibitors in herbivore resistance. Here, we identified a serine protease inhibitor, CsSERPIN1, from the tea plant (Camellia sinensis). Expression of CsSERPIN1 was not strongly affected by the assessed biotic and abiotic stresses. In vitro and in vivo experiments showed that CsSERPIN1 strongly inhibited the activities of digestive protease activities of trypsin and chymotrypsin. Transient or heterologous expression of CsSERPIN1 significantly reduced herbivory by two destructive herbivores, the tea geometrid and fall armyworm, in tea and Arabidopsis plants, respectively. The expression of CsSERPIN1 in Arabidopsis did not negatively influence the growth of the plants under the measured parameters. Our findings suggest that CsSERPIN1 can inactivate gut digestive proteases and suppress the growth and development of herbivores, making it a promising candidate for pest prevention in agriculture.

2.
Hortic Res ; 7(1): 52, 2020.
Article in English | MEDLINE | ID: mdl-32257238

ABSTRACT

Plants release large amounts of volatile organic compounds (VOCs) in response to attackers. Several VOCs can serve as volatile signals to elicit defense responses in undamaged tissues and neighboring plants, but many questions about the ecological functions of VOCs remain unanswered. Tea plants are impacted by two harmful invaders, the piercing herbivore Empoasca (Matsumurasca) onukii Matsuda and the pathogen Colletotrichum fructicola. To determine the VOC signals in tea, we confirmed CsOPR3 as a marker gene and set up a rapid screening method based on a 1.51 kb CsOPR3 promoter fused with a ß-glucuronidase (GUS) reporter construct (OPR3p::GUS) in Arabidopsis. Using this screening system, a terpenoid volatile (E)-nerolidol was identified as a potent signal that elicits plant defenses. The early responses triggered by (E)-nerolidol included the activation of a mitogen-activated protein kinase and WRKY, an H2O2 burst, and the induction of jasmonic acid and abscisic acid signaling. The induced plants accumulated high levels of defense-related chemicals, which possessed broad-spectrum anti-herbivore or anti-pathogen properties, and ultimately triggered resistance against Empoasca onukii and Colletotrichum fructicola in tea. We propose that these findings can supply an environmentally friendly management strategy for controlling an insect pest and a disease of tea plants.

3.
Physiol Plant ; 169(4): 529-543, 2020 Aug.
Article in English | MEDLINE | ID: mdl-32196677

ABSTRACT

Multiple isoforms of chitinases participate in plant defense against outside invaders. However, the functions of hydrolase family 19 (GH19) chitinases on pest control remain largely unknown. Here we reported the isolation and functional analysis of a gene CsChi19, which encodes a GH19 endochitinase protein of 332 amino acid residues from tea plant (Camellia sinensis). CsChi19 expression levels were upregulated in response to mechanical wounding, infestation by two important pests: the tea geometrid Ectropis grisescens and the tea green leafhopper Empoasca (Matsumurasca) onukii, a fungal pathogen Colletotrichum fructicola, and treatment with two phytohormones: jasmonic acid (JA) and salicylic acid. CsChi19 was heterologously expressed in Escherichia coli, and its catalytic function was further elucidated. The protein could hydrolyze colloidal chitin, and the optimum temperature and pH for its activity was 40°C and pH 5.0. CsChi19 were found to be toxic to tea pests when they were fed on artificial diets containing this protein. Interestingly, the regurgitant derived from E. grisescens fed with artificial diets containing CsChi19 protein induced stronger expression of CsMPK3, more JA burst, more accumulation of defense-related secondary metabolites, and more emission of volatiles than the regurgitant derived from E. grisescens fed only with artificial diets. Our results provide first evidence that CsChi19 is involved in mediating a novel defense mechanism of tea plant through altering the composition of the regurgitant.


Subject(s)
Camellia sinensis , Chitinases , Lepidoptera , Animals , Plant Growth Regulators , Tea
4.
Pest Manag Sci ; 76(4): 1304-1312, 2020 Apr.
Article in English | MEDLINE | ID: mdl-31595641

ABSTRACT

BACKGROUND: The tea leafhopper, Empoasca onukii Matsuda (Hemiptera: Cicadellidae), is a major pest of tea plants in China. Here, we evaluated the repellent properties of eight volatile chemicals alone and in various combinations as tools for the management of this pest in tea gardens. These chemicals were from the Alliaceae and other aromatic plants, and are known to repel various insect species. RESULTS: Among the eight volatile compounds, dimethyl disulfide (DMDS), 1,8-cineole and allyl methyl sulfide were significantly repellent towards E. onukii adults. DMDS and 1,8-cineole were mixed to formulate a binary repellent. Under field conditions, spraying and slow-release applications of the mixture significantly decreased the density of E. onukii adults. The repelling effect after spraying was very short, only ∼ 2 days, but the slow-release mixture had a longer term repelling effect on E. onukii adults. High emission of the slow-release mixture, which was achieved by increasing the number of slow-release bottles, had a stronger repellent effect than low emission. Moreover, when the amount emitted was sufficient, the slow-release mixture significantly decreased the number of leafhopper nymphs in a treated tea-plant line, and significantly decreased the number of leafhopper adults and nymphs in a tea-plant line adjacent to the treated area. CONCLUSION: This study demonstrates the repellent action of a mixture of DMDS and 1,8-cineole applied by a slow-release method against E. onukii in a tea plantation. This mixture has potential applications in integrated pest management schemes. © 2019 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.


Subject(s)
Camellia sinensis , Hemiptera , Animals , China , Nymph , Tea
5.
Insects ; 11(1)2019 Dec 23.
Article in English | MEDLINE | ID: mdl-31877916

ABSTRACT

Since the identification of the Ectropis grisescens sex pheromone, no effective control technology based on this pheromone has yet been developed and evaluated. In this study, pheromone proportion and dosage, sustained-release dispensers, and pheromone lure-matched traps were optimized. The mass trapping technology developed with the above optimized parameters was tested in a field trial. The results show that two compounds, (Z,Z,Z)-3,6,9-octadecatriene and (Z,Z)-3,9-cis-6,7-epoxy-octadecadiene, at a ratio of 30:70 and impregnated into rubber septa at 1 mg, were the most attractive to male moths. These compounds provided the best performance when combined with a sticky wing trap. Adult male moth monitoring data showed that there was a lower population density in the trapping plot compared with the control plot, and there was a clear difference during the peak adult occurrence of the first five insect generations in 2017. The effect of mass trapping on the larva population was investigated in 2018; the control efficiency reached 49.27% after trapping of one generation of adults and was further reduced to 67.16% after two successive adult moth generations, compared with the control plot. The results of the present study provide a scientific basis for the establishment of sex pheromone-based integrated pest management strategies.

6.
J Plant Physiol ; 243: 153047, 2019 Dec.
Article in English | MEDLINE | ID: mdl-31639538

ABSTRACT

Acyl-CoA oxidase (ACX; EC 1.3.3.6) plays a vital role in the biosynthesis of jasmonic acid (JA) in plant peroxisomes. We previously identified an herbivore-induced gene CsACX1 in tea plant (Camellia sinensis) and showed CsACX1 was involved in the wound-induced synthesis of jasmonic acid (JA). Here, another ACX gene CsACX3 was isolated from tea plant. CsACX3 was predicted to consist of 684 amino acid residues. CsACX3 can be induced by mechanical wounding, JA application, and infestation by the tea geometrid Ectropis obliqua Prout and the tea green leafhopper Empoasca (Matsumurasca) onukii Matsuda. These expression patterns are consistent with the previously reported expression pattern of CsACX1 under such treatments. Recombinant CsACX3 showed preference for medium-chain acyl-coA oxidase substrates (C8- to C14-CoA). CsACX3 expression could also be induced by the infection of a pathogen Colletotrichum gloeosporioides (Cgl), and the increased ACX activities in tea plants were correlated with the Cgl-induced CsACX3 expression. Cgl could not induce the expression of CsACX1, which showed preference for C12- to C16-CoA substrates. The constitutive expression of CsACX3 rescued wound-induced JA biosynthesis and enhanced the Cgl-induced JA biosynthesis in Arabidopsis mutant atacx1. However, constitutive expression of CsACX1 could not enhance the Cgl-induced JA biosynthesis in atacx1 plant. These results indicate that CsACX1 and CsACX3 functions overlap and have distinct roles in the wound- and pathogen-activated de novo JA synthesis via enzymatic routes that utilize different ACX isozymes in tea plant.


Subject(s)
Acyl-CoA Oxidase/genetics , Camellia sinensis/genetics , Cyclopentanes/metabolism , Gene Expression , Oxylipins/metabolism , Plant Proteins/genetics , Acyl-CoA Oxidase/metabolism , Amino Acid Sequence , Animals , Base Sequence , Camellia sinensis/enzymology , Camellia sinensis/metabolism , Colletotrichum/physiology , Feeding Behavior , Food Chain , Hemiptera/physiology , Isoenzymes/genetics , Isoenzymes/metabolism , Moths/physiology , Phylogeny , Plant Diseases/microbiology , Plant Proteins/chemistry , Plant Proteins/metabolism , Recombinant Proteins/genetics , Recombinant Proteins/metabolism
7.
J Plant Res ; 132(2): 285-293, 2019 Mar.
Article in English | MEDLINE | ID: mdl-30758750

ABSTRACT

Green leaf volatiles (GLVs) play a vital role in enhancing herbivore-associated defense responses, but the mechanism by which they precisely regulate such responses is not well understood. (Z)-3-Hexenol (z3HOL), an important component of GLVs, effectively activates the defense of tea plants (Camellia sinensis) against a tea geometrid (TG) Ectropis obliqua Prout. To elucidate the molecular mechanisms of defense activation by z3HOL, RNA-Sequencing was employed to investigate the effect of z3HOL on transcriptome responses to TG in tea plants. A total of 318 upregulated genes were identified, and expression of 10 unigenes was validated by quantitative real-time PCR. Among these 318 upregulated genes, 56 were defense-related, including 6 key enzyme genes in jasmonic acid, and ethylene biosynthesis, 24 signal transduction genes, and 12 insect-responsive transcription factors. Most of the defense-related genes are induced by JA, TG, or wounding treatments, suggesting that JA signaling plays a vital role in z3HOL-induced tea defense against TG.


Subject(s)
Camellia sinensis/metabolism , Herbivory , Moths , Transcriptome , Animals , Cyclopentanes/metabolism , Ethylenes/biosynthesis , Hexanols , Larva , Oxylipins/metabolism
8.
Sci Rep ; 9(1): 814, 2019 01 28.
Article in English | MEDLINE | ID: mdl-30692583

ABSTRACT

The tea plant (Camellia sinensis) suffers heavily from a harmful piercing pest, the tea green leafhopper (TLH) Empoasca (Matsumurasca) onukii Matsuda. In the present study, we studied the effect of an efficient elicitor of plant disease resistance, the ß-1,3-glucan laminarin, on the induced defense against TLH in tea plants. Defense responses elicited by laminarin in tea include the activation of mitogen-activated protein kinases and WRKY, the burst of H2O2, salicylic acid, and abscisic acid, and the accumulation of direct-defense chemicals (including chitinase, phenylalanine ammonia lyase, callose, polyphenol oxidase, and flavonol synthase), as well as the production of volatile compounds. The laminarin-treated tea plants reduced the performance of TLH and enhanced the attractiveness to the egg parasitoid wasp of TLH, Stethynium empoascae Subba Rao. In the field experiment, laminarin application effectively reduced the number of TLH by attracting parasitoids. These results suggest that laminarin can induce protection against TLH by regulating signaling pathways in tea plant. Our study also proposes an environment friendly strategy for the integrated management of an economically important piercing pest.


Subject(s)
Camellia sinensis/parasitology , Disease Resistance , Glucans/pharmacology , Hemiptera/pathogenicity , Plant Proteins/genetics , Abscisic Acid/metabolism , Animals , Camellia sinensis/drug effects , Camellia sinensis/genetics , Extracellular Signal-Regulated MAP Kinases/genetics , Gene Expression Regulation, Plant/drug effects , Hydrogen Peroxide/metabolism , Salicylic Acid/metabolism , Signal Transduction/drug effects , Transcription Factors/genetics , Volatile Organic Compounds/metabolism
9.
Plant Physiol Biochem ; 135: 132-140, 2019 Feb.
Article in English | MEDLINE | ID: mdl-30529979

ABSTRACT

The biosynthesis of jasmonic acid (JA) in plant peroxisomes requires the action of acyl-CoA oxidase (ACX; EC 1.3.3.6). Multiple isoforms of ACXs have been identified in various annual herbaceous plants, but the genes encoding these enzymes in perennial woody plants are yet to be fully investigated. In this study, an ACX gene named CsACX1 (GeneBank accession: KX650077.1) was isolated from tea plant (Camellia sinensis L.). CsACX1 was predicted to consist of 664 amino acid residues. Transcriptional analysis revealed that CsACX1 can be induced by mechanical wounding, JA application, and infestation by the tea geometrid Ectropis obliqua Prout and the tea green leafhopper Empoasca (Matsumurasca) onukii Matsuda. To further elucidate the function of CsACX1, it was heterologously expressed in a bacterial system and characterized. Recombinant CsACX1 showed preference for C12 ∼ C16-CoA substrates. The constitutive expression of CsACX1 can rescue wound-related JA biosynthesis in Arabidopsis mutant acx1. CsACX1 was expressed in different organs, predominantly in flowers. Notably, CsACX1 transcripts were detected up-regulated during flower opening, and the JA levels were correlated with CsACX1 expression. All these results enrich our knowledge of the regulatory pathway involved in the JA biosynthesis in tea, and helps further understand the defense mechanism of tea plant against insects.


Subject(s)
Acyl-CoA Oxidase/physiology , Camellia sinensis/physiology , Cyclopentanes/metabolism , Flowers/metabolism , Genes, Plant/physiology , Oxylipins/metabolism , Plant Growth Regulators/metabolism , Plant Proteins/physiology , Acyl-CoA Oxidase/genetics , Animals , Arabidopsis , Camellia sinensis/genetics , Camellia sinensis/metabolism , Escherichia coli , Flowers/growth & development , Flowers/physiology , Gene Expression Regulation, Plant , Genes, Plant/genetics , Hemiptera , Herbivory , Phylogeny , Plant Growth Regulators/physiology , Plant Proteins/genetics , Plants, Genetically Modified , Real-Time Polymerase Chain Reaction , Sequence Alignment , Sequence Analysis, DNA
10.
J Econ Entomol ; 112(1): 277-283, 2019 02 12.
Article in English | MEDLINE | ID: mdl-30508146

ABSTRACT

Ectropis grisescens Warren 1894 (Lepidopotera: Geometridae) and Ectropis obliqua Prout 1915 (Lepidopotera: Geometridae) are the most destructive chewing pests in China's tea plantations. Ectropis grisescens sex pheromone lures and E. obliqua nucleopolyhedrosis virus (EoNPV) are two species-specific and effective bio-control technologies to control these pests. Because these two species are morphologically similar, tea growers are unable to discriminate them by visual inspection. Hence, determining whether to use E. grisescens sex pheromone lures or EoNPV is difficult without knowledge on the geographical distribution of these two Ectropis species in China. In this study, we developed an efficient identification method, which is considerably cheaper and faster than sequencing the cytochrome c oxidase I gene. Overall, 2,588 E. grisescens and E. obliqua samples, collected from 13 provinces and municipalities in China covering the major regions where these pests have been reported, were identified. All insect samples from southern Jiangsu Province were identified as E. obliqua. Both Ectropis species were mix-distributed at the Anhui-Zhejiang Province border areas, whereas E. grisescens was mostly collected from the other sampling sites. Thus, E. obliqua might be mainly distributed at the junction of Jiangsu, Anhui, and Zhejiang Provinces. In contrast, E. grisescens has a considerably wide distribution area and is the main lepidopteran pest in the tea plantations of China. Our results contribute to improve the management of E. grisescens and E. obliqua populations and provide new insights for further studies on these two species.


Subject(s)
Moths/classification , Animals , Base Sequence , China , Geography , Moths/genetics , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length
11.
Front Physiol ; 9: 1602, 2018.
Article in English | MEDLINE | ID: mdl-30487755

ABSTRACT

The sibling species Ectropis grisescens and E. obliqua are the major chewing tea pests in China. A difference in sex pheromone components plays a central role in premating isolation in these two species. To investigate the mechanism of premating isolation in these two Ectropis species, we sequenced the transcriptomes of the antennae of female and male E. obliqua individuals and performed phylogenetic analyses, abundance analyses, and tissue expression profile analyses to compare the olfactory genes involved in the detection of sex pheromones. A total of 36 odorant-binding proteins (OBPs) and 52 olfactory receptors (ORs) were identified in E. obliqua. Phylogenetic analyses showed that EoblOBP2, 3, and 25 were grouped in the pheromone-binding protein clade with EgriOBP2, 3, 25, and another lepidopteran PBP. EoblOR25 and 28 were grouped with EgriOR25, 28, and pheromone receptors for the detection of Type-I sex pheromone components. EoblOR24, 31, 37, and 44 were grouped with EgriOR24, 31, 37, and 44. All of these 4 EoblORs and 4 EgriORs showed higher abundance in male antennae than in female ones. Therefore, OBP2, 3, 25 and OR24, 31, 37, 44 of E. grisescens and E. obliqua might be responsible for sex pheromone component detection. However, the sequences of these genes in E. grisescens and E. obliqua were more than 90% identical. This indicates that these orthologous genes might play similar roles in the detection of sex pheromones. In contrast, the observed OBPs and ORs differed in abundance between the antennae of the two Ectropis species. Therefore, we speculate that these two Ectropis species use the different transcript levels of PRs to differentiate sex pheromone components. The results of the present study might contribute in deciphering the mechanism for premating isolation in these species and may be of use in devising strategies for their management.

12.
G3 (Bethesda) ; 8(3): 899-908, 2018 03 02.
Article in English | MEDLINE | ID: mdl-29317471

ABSTRACT

Moths can biosynthesize sex pheromones in the female sex pheromone glands (PGs) and can distinguish species-specific sex pheromones using their antennae. However, the biosynthesis and transportation mechanism for Type II sex pheromone components has rarely been documented in moths. In this study, we constructed a massive PG transcriptome database (14.72 Gb) from a moth species, Ectropis grisescens, which uses type II sex pheromones and is a major tea pest in China. We further identified putative sex pheromone biosynthesis and transportation-related unigenes: 111 cytochrome P450 monooxygenases (CYPs), 25 odorant-binding proteins (OBPs), and 20 chemosensory proteins (CSPs). Tissue expression and phylogenetic tree analyses showed that one CYP (EgriCYP341-fragment3), one OBP (EgriOBP4), and one CSP (EgriCSP10) gene displayed an enriched expression in the PGs, and that EgriOBP2, 3, and 25 are clustered in the moth pheromone-binding protein clade. We considered these our candidate genes. Our results yielded large-scale PG sequence information for further functional studies.


Subject(s)
Gene Expression Profiling , Genes, Insect , Moths/genetics , Moths/metabolism , Sex Attractants/metabolism , Animals , Biological Transport , Camellia sinensis/parasitology , Computational Biology/methods , Female , Gene Expression Regulation , High-Throughput Nucleotide Sequencing , Male , Molecular Sequence Annotation , Moths/classification , Organ Specificity , Phylogeny , Sex Attractants/biosynthesis , Transcriptome
13.
J Econ Entomol ; 111(2): 629-636, 2018 04 02.
Article in English | MEDLINE | ID: mdl-29361007

ABSTRACT

The tea leafhopper, Empoasca onukii Matsuda, is a serious pest of the tea plant. E. onukii prefers to inhabit vigorously growing tender tea leaves. The host selection of E. onukii adults may be associated with plant volatile compounds (VOCs). We sought to identify potentially attractive VOCs from tea leaves at three different ages and test the behavioral responses of E. onukii adults to synthetic VOC blends in the laboratory and field to aid in developing an E. onukii adult attractant. In darkness, the fresh or mature tea leaves of less than 1-mo old could attract more leafhoppers than the mature branches (MB) that had many older leaves (leaf age >1 mo). Volatile analysis showed that the VOC composition of the fresh leaves was the same as that of the mature leaves, but linalool and indole were not at detectable levels in VOCs from the MB. Moreover, the mass ratio differed for each common volatile in the three types of tea leaves. When under competition with volatiles from the MB, the leafhoppers showed no significant tropism to each single volatile but could be attracted by the synthetic volatile blend imitating the fresh leaves. With the removal of some volatile components, the effective synthetic volatile blend was mixed with (Z)-3-hexen-1-ol, (Z)-3-hexenyl acetate, and linalool at a mass ratio of 0.6:23:12.6. These three volatiles may be the key components for the host selection of E. onukii adults and could be used as an attractant in tea gardens.


Subject(s)
Behavior, Animal , Camellia sinensis/chemistry , Hemiptera , Volatile Organic Compounds/analysis , Animals
14.
Front Physiol ; 8: 953, 2017.
Article in English | MEDLINE | ID: mdl-29209233

ABSTRACT

Tea grey geometrid (Ectropis grisescens), a devastating chewing pest in tea plantations throughout China, produces Type-II pheromone components. Little is known about the genes encoding proteins involved in the perception of Type-II sex pheromone components. To investigate the olfaction genes involved in E. grisescens sex pheromones and plant volatiles perception, we sequenced female and male antennae transcriptomes of E. grisescens. After assembly and annotation, we identified 153 candidate chemoreception genes in E. grisescens, including 40 odorant-binding proteins (OBPs), 30 chemosensory proteins (CSPs), 59 odorant receptors (ORs), and 24 ionotropic receptors (IRs). The results of phylogenetic, qPCR, and mRNA abundance analyses suggested that three candidate pheromone-binding proteins (EgriOBP2, 3, and 25), two candidate general odorant-binding proteins (EgriOBP1 and 29), six pheromone receptors (EgriOR24, 25, 28, 31, 37, and 44), and EgriCSP8 may be involved in the detection of Type-II sex pheromone components. Functional investigation by heterologous expression in Xenopus oocytes revealed that EgriOR31 was robustly tuned to the E. grisescens sex pheromone component (Z,Z,Z)-3,6,9-octadecatriene and weakly to the other sex pheromone component (Z,Z)-3,9-6,7-epoxyoctadecadiene. Our results represent a systematic functional analysis of the molecular mechanism of olfaction perception in E. grisescens with an emphasis on gene encoding proteins involved in perception of Type-II sex pheromones, and provide information that will be relevant to other Lepidoptera species.

15.
J Chem Ecol ; 43(6): 557-562, 2017 Jun.
Article in English | MEDLINE | ID: mdl-28601939

ABSTRACT

Insect cuticular hydrocarbons (CHCs) play important roles in chemical communication, as well as having ecological and physiological roles. The use of CHCs for mate recognition has been shown in many insect genera, but little is known about their use in the tea weevil Myllocerinus aurolineatus. Here, we provide evidence that CHCs on the surface of sexually mature M. aurolineatus females act as contact sex pheromones, facilitating mate recognition and eliciting copulatory behavior in male weevils. Using gas chromatography-mass spectrometry, we identify n-pentacosane and n-heptacosane as two potential contact pheromone components. Results from arena bioassays showed that n-pentacosane is a component of a contact pheromone of M. aurolineatus. Further results from the Y-tube olfactometer bioassays showed that n-pentacosane also acts as a volatile attractant. Our results greatly improve our understanding of the chemical ecology of M. aurolineatus.


Subject(s)
Alkanes/chemistry , Alkanes/metabolism , Sex Attractants/chemistry , Sex Attractants/physiology , Weevils/chemistry , Weevils/physiology , Animals , Female , Gas Chromatography-Mass Spectrometry , Male , Olfactometry , Sexual Behavior, Animal , Volatile Organic Compounds/chemistry
16.
Gene ; 615: 18-24, 2017 Jun 05.
Article in English | MEDLINE | ID: mdl-28322995

ABSTRACT

12-Oxophytodienoate reductase (OPR) is a key enzyme in the biosynthesis of jasmonic acid (JA), which plays an important role in plant defense responses. Although multiple isoforms of OPRs have been identified in various annual herbaceous plants, genes encoding these enzymes in perennial woody plants have yet to be fully investigated. In the tea plant, Camellia sinensis (L.), no OPR genes have been isolated, and their possible roles in tea plant development and defense mechanism remain unknown. In this study, a putative OPR gene, designated as CsOPR3, was isolated from tea plants for the first time through the rapid amplification of cDNA ends. The open reading frame of CsOPR3 is 1197bp in length, and encodes a protein of 398 amino acids. Real-time qPCR analysis revealed that CsOPR3 was expressed in different organs. In particular, CsOPR3 was highly expressed in flowers, leaves and stems but was weakly expressed in roots and seeds. CsOPR3 expression could be rapidly induced by mechanical wounding, and increased JA levels were correlated with the wound-induced CsOPR3 expression. The infestation of the tea geometrid (TG) Ectropis obliqua Prout, regurgitant derived from TG and exogenous JA application could enhance the CsOPR3 expression. Our study is the first to report that CsOPR3 plays an important role in JA biosynthesis and tea plant defense against herbivorous insects.


Subject(s)
Camellia sinensis/physiology , Herbivory , Oxidoreductases Acting on CH-CH Group Donors/genetics , Plant Proteins/genetics , Animals , Camellia sinensis/drug effects , Camellia sinensis/genetics , Cloning, Molecular , Cyclopentanes/metabolism , Cyclopentanes/pharmacology , Gene Expression Regulation, Plant , Moths , Oxidoreductases Acting on CH-CH Group Donors/metabolism , Oxylipins/metabolism , Oxylipins/pharmacology , Plant Proteins/metabolism
17.
Environ Entomol ; 44(5): 1425-33, 2015 Oct.
Article in English | MEDLINE | ID: mdl-26314008

ABSTRACT

The lacewing Chrysopa phyllochroma Waesmael is a polyphagous predator of many pests. Releasing lacewings is an important component of biological control programs, but it is difficult to establish populations on field crops. Electrophysiological and behavioral responses to 10 common plant volatiles were recorded to screen for lacewing-attracting compounds. Electroantennographic assays indicated that all of the tested compounds elicited responses from C. phyllochroma. Three green-leaf volatiles-(E)-2-hexenal, (Z)-3-hexenyl acetate, and (Z)-3-hexenol-produced the strongest responses. Weaker responses were observed to six terpenes-ocimene, linalool, (3E)-4,8-dimethyl-1,3,7-nonatriene, (E,E)-α-farnesene, limonene, and nerolidol-and to methyl salicylate. Using a Y-tube olfactometer, the behavioral assays of the eight most active compounds demonstrated that four-(Z)-3-hexenyl acetate, (Z)-3-hexenol, (3E)-4,8-dimethyl-1,3,7-nonatriene, and linalool-were significant attractants for C. phyllochroma at specific concentrations. Three common plant volatile compounds-(Z)-3-hexenyl acetate, (3E)-4,8-dimethyl-1,3,7-nonatriene, and linalool-were also found to significantly enhance female ovipositing, resulting in a concentration of eggs. These observations are important for lacewing release as a pest control measure because they suggest means for retaining individuals and establishing populations using common plant volatiles.


Subject(s)
Insecta/physiology , Plants/chemistry , Terpenes/pharmacology , Volatile Organic Compounds/pharmacology , Animals , Behavior, Animal/drug effects , Biological Control Agents , Electrophysiological Phenomena , Female , Odorants , Oviposition/physiology , Plant Leaves/chemistry , Sense Organs/physiology
18.
Pest Manag Sci ; 71(1): 96-104, 2015 Jan.
Article in English | MEDLINE | ID: mdl-24616107

ABSTRACT

BACKGROUND: A plant-based 'push-pull' strategy for Ectropis obliqua (Prout) (Lepidoptera: Geometridae) is being developed using semiochemicals in the volatiles of Rosmarinus officinalis (Lamiaceae). The aim of this study was to identify and quantify the bioactive components within R. officinalis by gas chromatography-electroantennographic detection (GC-EAD) and gas chromatography-mass spectrometry (GC-MS), and to test the antennal and behavioural responses of E. obliqua to these chemicals. The emission dynamics of bioactive chemicals was also monitored. RESULTS: GC-EAD experiments indicated that E. obliqua antennae responded to the following volatile compounds from R. officinalis: myrcene, α-terpinene, γ-terpinene, linalool, cis-verbenol, camphor, α-terpineol and verbenone, which were the minor constituents. Based on the dose-dependent antennal and behavioural responses of E. obliqua to these bioactive compounds, myrcene, γ-terpinene, linalool, cis-verbenol, camphor and verbenone were found to play a key role in repelling the moths, and the mixture that included all eight compounds was significantly more effective. The maximum emissions of these semiochemicals occurred at nightfall. CONCLUSIONS: The specifically bioactive compounds in R. officinalis volatiles are responsible for repelling E. obliqua adults. Results indicate that R. officinalis should be considered as a potential behaviour-modifying stimulus for 'push' components when developing 'push-pull' strategies for control of E. obliqua using semiochemicals.


Subject(s)
Arthropod Antennae/drug effects , Insect Repellents/pharmacology , Moths/drug effects , Rosmarinus/chemistry , Volatile Organic Compounds/pharmacology , Animals , Behavior, Animal/drug effects , Electrophysiological Phenomena , Female , Gas Chromatography-Mass Spectrometry , Insect Repellents/analysis , Male , Olfactometry , Photoperiod
19.
J Chem Ecol ; 40(10): 1080-9, 2014 Oct.
Article in English | MEDLINE | ID: mdl-25378120

ABSTRACT

Herbivore-induced plant volatiles have been reported to play a role in the host-searching behavior of herbivores. However, next to nothing is known about the effect of volatiles emitted from tea plants infested by Ectropis obliqua larvae on the behavior of conspecific adults. Here, we found that tea plants infested by E. obliqua caterpillars for 24 h were more attractive to both virgin male and female E. obliqua adults than were intact, uninfested tea plants; moreover, mated female E. obliqua moths were more attracted by infested tea plants and preferentially oviposited on these plants, whereas male moths were repelled by infested plants once they had mated. Volatile analysis revealed that the herbivore infestation dramatically increased the emission of volatiles. Among these volatiles, 17 compounds elicited antennal responses from both male and female virginal moths. Using a Y-tube olfactometer, we found that 3 of the 17 chemicals, benzyl alcohol, (Z)-3-hexenyl hexanoate, and (Z)-3-hexenal, were attractive, but two compounds, linalool and benzyl nitril, were repellent to virgin male and female moths. One chemical, (Z)-3-hexenyl acetate, was attractive only to virgin males. Mated females were attracted by three compounds, (Z)-3-hexenyl hexanoate, (Z)-3-hexenyl acetate, and (Z)-3-hexenal; whereas mated males were repelled by (Z)-3-hexenol. The findings provide new insights into the interaction between tea plants and the herbivores, and may help scientists develop new measures with which to control E. obliqua.


Subject(s)
Appetitive Behavior , Herbivory , Moths/physiology , Tea/physiology , Tea/parasitology , Volatile Organic Compounds/metabolism , Animals , Female , Host-Parasite Interactions , Larva/physiology , Male , Moths/anatomy & histology , Oviposition
20.
J Plant Res ; 127(4): 565-72, 2014 Jul.
Article in English | MEDLINE | ID: mdl-24888389

ABSTRACT

We investigated the effect of the SHAM treatment of tea plants on their induced defense on a tea geometrid (TG), Ectropis obliqua Prout. Treatment of tea leaves with SHAM reduced the performance of TG and TG-elicited level of the lipoxygenase gene CsiLOX1 and the putative allene oxide synthase gene CsiAOS1. The release of wound-induced green leaf volatiles (GLVs) and the expression of the hydroperoxide lyase (HPL) gene CsiHPL1 were also reduced by SHAM treatment. The negative effect of SHAM dramatically reduced the total hebivore-induced plant volatiles (HIPVs) and the attractiveness to the parasitoid wasp Apanteles sp. These results indicated that SHAM may negatively mediate tea defense response against TG by modulating the wound-induced emission of GLVs, the expression of genes involved in oxylipin pathway, and the emission of other HIPV compounds that mediate direct and indirect defenses.


Subject(s)
Camellia sinensis/drug effects , Herbivory/drug effects , Moths/physiology , Salicylamides/pharmacology , Aldehyde-Lyases/genetics , Animals , Camellia sinensis/enzymology , Camellia sinensis/genetics , Camellia sinensis/parasitology , Cyclopentanes/metabolism , Cytochrome P-450 Enzyme System/genetics , Female , Food Chain , Gas Chromatography-Mass Spectrometry , Gene Expression Regulation, Plant/drug effects , Moths/parasitology , Oxylipins/metabolism , Plant Leaves/drug effects , Plant Leaves/enzymology , Plant Proteins/genetics , Plant Proteins/metabolism , Real-Time Polymerase Chain Reaction , Signal Transduction , Volatile Organic Compounds/metabolism , Wasps/physiology
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