Functional Characterization of CaSpr2 in Jasmonate-Dependent Induced Defense Against Western Flower Thrips in Capsicum annuum
Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Insects and Plants
2.2. Sequence Comparisons and Phylogenetic Analyses
2.3. TRV-Mediated Silencing Assays
2.4. RNA Isolation and RT-qPCR
2.5. Hormone Quantification
2.6. Selection Preference of WFTs for Host Plants
2.7. The Life Tables of WFT Feeding on TRV-GFP and TRV-CaSpr2-Infected Pepper Plants
3. Results
3.1. Identification and Silencing of CaSpr2 in Pepper
3.2. Silencing of CaSpr2 Exerts a Negative Impact on JA Accumulation and Attracts the Feeding of WFTs
3.3. Silencing of CaSpr2 Is Beneficial for the Developmental Durations of WFTs
3.4. The Survival Rates of WFTs Reared on CaSpr2-Silenced Leaves Significantly Increased
3.5. The Life Expectancy and Reproductive Values of WFTs Fed on TRV-CaSpr2-Infected Pepper Leaves Are Prolonged
3.6. WFTs Demonstrates Better Population Parameters When Feeding on CaSpr2-Silenced Leaves
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Kessler, A.; Baldwin, I.T. Plant responses to insect herbivory: The emerging molecular analysis. Annu. Rev. Plant Biol. 2002, 53, 299–328. [Google Scholar] [CrossRef]
- Reitz, S.R. Biology and ecology of the western flower thrips (Thysanoptera: Thripidae): The making of a pest. Fla. Entomol. 2009, 92, 7–13. [Google Scholar] [CrossRef]
- Steenbergen, M.; Abd-El-Haliem, A.; Bleeker, P.; Dicke, M.; Escobar-Bravo, R.; Cheng, G.; Haring, M.A.; Kant, M.R.; Kappers, I.; Klinkhamer, P.G.L.; et al. Thrips advisor: Exploiting thrips-induced defences to combat pests on crops. J. Exp. Bot. 2018, 69, 1837–1848. [Google Scholar] [CrossRef]
- Maharijaya, A.; Vosman, B.; Steenhuis-Broers, G.; Harpenas, A.; Purwito, A.; Visser, R.G.F.; Voorrips, R.E. Screening of pepper accessions for resistance against two thrips species (Frankliniella occidentalis and Thrips parvispinus). Euphytica 2011, 177, 401–410. [Google Scholar] [CrossRef]
- Gera, A.; Kritzman, A.; Cohen, J.; Raccah, B. First report of impatiens necrotic spot tospovirus (INSV) in Israel. Plant Dis. 1999, 83, 587. [Google Scholar] [CrossRef]
- Maris, P.C.; Joosten, N.N.; Peters, D.; Goldbach, R.W. Thrips resistance in pepper and its consequences for the acquisition and inoculation of tomato spotted wilt virus by the western flower thrips. Phytopathology 2003, 93, 96–101. [Google Scholar] [CrossRef]
- Bielza, P. Insecticide resistance management strategies against the western flower thrips, Frankliniella occidentalis. Pest Manag. Sci. 2008, 64, 1131–1138. [Google Scholar] [CrossRef] [PubMed]
- Mithöfer, A.; Boland, W. Plant defense against herbivores: Chemical aspects. Annu. Rev. Plant Biol. 2012, 63, 431–450. [Google Scholar] [CrossRef]
- Wu, J.; Baldwin, I.T. New insights into plant responses to the attack from insect herbivores. Annu. Rev. Genet. 2010, 44, 1–24. [Google Scholar] [CrossRef] [PubMed]
- Erb, M.; Robert, C.A. Sequestration of plant secondary metabolites by insect herbivores: Molecular mechanisms and ecological consequences. Curr. Opin. Insect Sci. 2016, 14, 8–11. [Google Scholar] [CrossRef] [PubMed]
- Fernández de Bobadilla, M.; Vitiello, A.; Erb, M.; Poelman, E.H. Plant defense strategies against attack by multiple herbivores. Trends Plant Sci. 2022, 27, 528–535. [Google Scholar] [CrossRef]
- Bari, R.; Jones, J.D. Role of plant hormones in plant defence responses. Plant Mol. Biol. 2009, 69, 473–488. [Google Scholar] [CrossRef]
- Hickman, R.; Van Verk, M.C.; Van Dijken, A.J.H.; Mendes, M.P.; Vroegop-Vos, I.A.; Caarls, L.; Steenbergen, M.; Van der Nagel, I.; Wesselink, G.J.; Jironkin, A.; et al. Architecture and dynamics of the jasmonic acid gene regulatory network. Plant Cell 2017, 29, 2086–2105. [Google Scholar] [CrossRef]
- Santino, A.; Taurino, M.; De Domenico, S.; Bonsegna, S.; Poltronieri, P.; Pastor, V.; Flors, V. Jasmonate signaling in plant development and defense response to multiple (a)biotic stresses. Plant Cell Rep. 2013, 32, 1085–1098. [Google Scholar] [CrossRef]
- Schuman, M.C.; Baldwin, I.T. The layers of plant responses to insect herbivores. Annu. Rev. Entomol. 2016, 61, 373–394. [Google Scholar] [CrossRef] [PubMed]
- Parde, V.D.; Sharma, H.C.; Kachole, M.S. In vivo inhibition of Helicoverpa armigera gut pro-proteinase activation by non-host plant protease inhibitors. J. Insect Physiol. 2010, 56, 1315–1324. [Google Scholar] [CrossRef]
- Scott, I.M.; Thaler, J.S.; Scott, J.G. Response of a generalist herbivore Trichoplusia ni to jasmonate-mediated induced defense in tomato. J. Chem. Ecol. 2010, 36, 490–499. [Google Scholar] [CrossRef]
- Shivaji, R.; Camas, A.; Ankala, A.; Engelberth, J.; Tumlinson, J.H.; Williams, W.P.; Wilkinson, J.R.; Luthe, D.S. Plants on constant alert: Elevated levels of jasmonic acid and jasmonate-induced transcripts in caterpillar-resistant maize. J. Chem. Ecol. 2010, 36, 179–191. [Google Scholar] [CrossRef]
- Abe, H.; Ohnishi, J.; Narusaka, M.; Seo, S.; Narusaka, Y.; Tsuda, S.; Kobayashi, M. Function of jasmonate in response and tolerance of Arabidopsis to thrip feeding. Plant Cell Physiol. 2008, 49, 68–80. [Google Scholar] [CrossRef] [PubMed]
- Egger, B.; Koschier, E.H. Behavioural responses of Frankliniella occidentalis Pergande larvae to methyl jasmonate and cis-jasmone. J. Pest Sci. 2014, 87, 53–59. [Google Scholar] [CrossRef] [PubMed]
- Abe, H.; Shimoda, T.; Ohnishi, J.; Kugimiya, S.; Narusaka, M.; Seo, S.; Narusaka, Y.; Tsuda, S.; Kobayashi, M. Jasmonate-dependent plant defense restricts thrips performance and preference. BMC Plant Biol. 2009, 9, 97. [Google Scholar] [CrossRef]
- Chen, G.; Kim, H.K.; Klinkhamer, P.G.; Escobar-Bravo, R. Site-dependent induction of jasmonic acid-associated chemical defenses against western flower thrips in Chrysanthemum. Planta 2019, 251, 8. [Google Scholar] [CrossRef]
- Omer, A.D.; Granett, J.; Karban, R.; Villa, E.M. Chemically-induced resistance against multiple pests in cotton. Int. J. Pest Manag. 2001, 47, 49–54. [Google Scholar] [CrossRef]
- Escobar-Bravo, R.; Klinkhamer, P.G.L.; Leiss, K.A. Induction of jasmonic acid-associated defenses by thrips alters host suitability for conspecifics and correlates with increased trichome densities in tomato. Plant Cell Physiol. 2017, 58, 622–634. [Google Scholar] [CrossRef]
- Li, C.Y.; Liu, G.H.; Xu, C.C.; Lee, G.I.; Bauer, P.; Ling, H.Q.; Ganal, M.W.; Howe, G.A. The tomato suppressor of prosystemin-mediated responses2 gene encodes a fatty acid desaturase required for the biosynthesis of jasmonic acid and the production of a systemic wound signal for defense gene expression. Plant Cell 2003, 15, 1646–1661. [Google Scholar] [CrossRef] [PubMed]
- Avila, C.A.; Arévalo-Soliz, L.M.; Jia, L.; Navarre, D.A.; Chen, Z.; Howe, G.A.; Meng, Q.-W.; Smith, J.E.; Goggin, F.L. Loss of function of FATTY ACID DESATURASE7 in tomato enhances basal aphid resistance in a salicylate-dependent manner. Plant Physiol. 2012, 158, 2028–2041. [Google Scholar] [CrossRef] [PubMed]
- Casarrubias-Castillo, K.; Montero-Vargas, J.M.; Dabdoub-González, N.; Winkler, R.; Martinez-Gallardo, N.A.; Zañudo-Hernández, J.; Avilés-Arnaut, H.; Délano-Frier, J.P. Distinct gene expression and secondary metabolite profiles in suppressor of prosystemin-mediated responses2 (spr2) tomato mutants having impaired mycorrhizal colonization. PeerJ 2020, 8, e8888. [Google Scholar] [CrossRef] [PubMed]
- Li, L.; Li, C.; Lee, G.I.; Howe, G.A. Distinct roles for jasmonate synthesis and action in the systemic wound response of tomato. Proc. Natl. Acad. Sci. USA 2002, 99, 6416–6421. [Google Scholar] [CrossRef]
- Sánchez-Hernández, C.; López, M.G.; Délano-Frier, J.P. Reduced levels of volatile emissions in jasmonate-deficient spr2 tomato mutants favour oviposition by insect herbivores. Plant Cell Environ. 2006, 29, 546–557. [Google Scholar] [CrossRef]
- Aiyar, A. The use of CLUSTAL W and CLUSTAL X for multiple sequence alignment. Methods Mol. Biol. 2000, 132, 221–241. [Google Scholar] [CrossRef]
- Kumar, S.; Stecher, G.; Tamura, K. MEGA7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol. Biol. Evol. 2016, 33, 1870–1874. [Google Scholar] [CrossRef]
- Liu, Y.L.; Schiff, M.; Dinesh-Kumar, S.P. Virus-induced gene silencing in tomato. Plant J. 2002, 31, 777–786. [Google Scholar] [CrossRef]
- Liu, Y.L.; Schiff, M.; Marathe, R.; Dinesh-Kumar, S.P. Tobacco Rar1, EDS1 and NPR1/NIM1 like genes are required for N-mediated resistance to tobacco mosaic virus. Plant J. 2002, 30, 415–429. [Google Scholar] [CrossRef]
- Ratcliff, F.; Martin-Hernandez, A.M.; Baulcombe, D.C. Tobacco rattle virus as a vector for analysis of gene function by silencing. Plant J. 2001, 25, 237–245. [Google Scholar] [CrossRef]
- Almeida Trapp, M.; De Souza, G.D.; Rodrigues-Filho, E.; Boland, W.; Mithöfer, A. Validated method for phytohormone quantification in plants. Front. Plant Sci. 2014, 5, 417. [Google Scholar] [CrossRef] [PubMed]
- Li, H.; Chen, Y.X.; Lu, C.C.; Tian, H.J.; Lin, S.; Wang, L.; Linghu, T.T.; Zheng, X.; Wei, H.; Fan, X.J.; et al. Chemosensory protein regulates the behavioural response of Frankliniella intonsa and Frankliniella occidentalis to tomato zonate spot virus-infected pepper (Capsicum annuum). PLoS Pathog. 2023, 19, e1011380. [Google Scholar] [CrossRef]
- Chi, H. Life-table analysis incorporating both sexes and variable development rates among individuals. Environ. Entomol. 1988, 17, 26–34. [Google Scholar] [CrossRef]
- Chi, H.; Liu, H.S.I. Two new methods for study of insect population ecology. Bull. Inst. Zool. Acad. Sin. 1984, 24, 225–240. [Google Scholar]
- Li, D.; Zhi, J.; Yue, W.; Zhang, T.; Liu, L. Resistance to spinetoram affects host adaptability of Frankliniella occidentalis (Thysanoptera: Thripidae) based on detoxifying enzyme activities and an age-stage-two-sex life table. Environ. Entomol. 2022, 51, 780–789. [Google Scholar] [CrossRef] [PubMed]
- Chi, H.; You, M.; Atlıhan, R.; Smith, C.L.; Kavousi, A.; Özgökçe, M.S.; Güncan, A.; Tuan, S.-J.; Fu, J.-w.; Xu, Y.-y.; et al. Age-Stage, two-sex life table: An introduction to theory, data analysis, and application. Entomol. Gen. 2020, 40, 103–124. [Google Scholar] [CrossRef]
- Chung, E.; Seong, E.; Kim, Y.C.; Chung, E.J.; Oh, S.K.; Lee, S.; Park, J.M.; Joung, Y.H.; Choi, D. A method of high frequency virus-induced gene silencing in chili pepper (Capsicum annuum L. cv. Bukang). Mol. Cells 2004, 17, 377–380. [Google Scholar] [CrossRef]
- Li, M.Y.; Yu, G.H.; Cao, C.L.; Liu, P. Metabolism, signaling, and transport of jasmonates. Plant Commun. 2021, 2, 100231. [Google Scholar] [CrossRef]
- Howe, G.A.; Ryan, C.A. Suppressors of systemin signaling identify genes in the tomato wound response pathway. Genetics 1999, 153, 1411–1421. [Google Scholar] [CrossRef]
- Sun, J.Q.; Jiang, H.L.; Li, C.Y. Systemin/jasmonate-mediated systemic defense signaling in tomato. Mol. Plant 2011, 4, 607–615. [Google Scholar] [CrossRef] [PubMed]
- Lee, G.I.; Howe, G.A. The tomato mutant spr1 is defective in systemin perception and the production of a systemic wound signal for defense gene expression. Plant J. 2003, 33, 567–576. [Google Scholar] [CrossRef] [PubMed]
- Li, C.; Zhao, J.; Jiang, H.; Wu, X.; Sun, J.; Zhang, C.; Wang, X.; Lou, Y.; Li, C. The wound response mutant suppressor of prosystemin-mediated responses6 (spr6) is a weak allele of the tomato homolog of CORONATINE-INSENSITIVE1 (COI1). Plant Cell Physiol. 2006, 47, 653–663. [Google Scholar] [CrossRef] [PubMed]
- Zhao, X.Q.; He, Y.Q.; Liu, Y.X.; Wang, Z.F.; Zhao, J. JAZ proteins: Key regulators of plant growth and stress response. Crop J. 2024, 12, 1505–1516. [Google Scholar] [CrossRef]



| Parameter | TRV-GFP | TRV-CaSpr2 | |||
|---|---|---|---|---|---|
| n | Mean ± SE | n | Mean ± SE | ||
| First instar larval stage (d) | 99 | 1.000 ± 0.000 | 93 | 0.973 ± 0.012 | * |
| Second instar larval stage (d) | 90 | 4.428 ± 0.081 | 88 | 4.620 ± 0.083 | * |
| Prepupal stage (d) | 88 | 1.006 ± 0.017 | 88 | 0.989 ± 0.018 | ns |
| Pupal stage (d) | 88 | 2.460 ± 0.048 | 88 | 2.403 ± 0.048 | ns |
| Preadult duration (d) | 88 | 12.869 ± 0.084 | 88 | 13.012 ± 0.079 | ns |
| Preadult survival rate | 99 | 0.889 ± 0.032 | 93 | 0.946 ± 0.023 | ns |
| Female longevity (d) | 40 | 12.296 ± 1.379 | 41 | 17.463 ± 0.889 | * |
| Male longevity (d) | 48 | 9.135 ± 0.536 | 47 | 12.138 ± 0.762 | * |
| Total preoviposition period (d) | 31 | 15.097 ± 0.249 | 41 | 14.976 ± 0.162 | ns |
| Fecundity | 40 | 48.643 ± 7.747 | 41 | 79.584 ± 5.922 | * |
| Population Parameter | TRV-GFP | TRV-CaSpr2 | |
|---|---|---|---|
| r (d−1) | 0.143 ± 0.011 | 0.174 ± 0.008 | * |
| λ (d−1) | 1.154 ± 0.012 | 1.190 ± 0.009 | * |
| R0 (offspring/individual) | 19.661 ± 3.932 | 35.105 ± 4.856 | * |
| T (d) | 20.701 ± 0.510 | 20.393 ± 0.323 | ns |
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Chen, X.; Lin, S.; Linghu, T.; Yu, Y.; Li, H.; Chen, Y.; Wei, H.; Chen, Y. Functional Characterization of CaSpr2 in Jasmonate-Dependent Induced Defense Against Western Flower Thrips in Capsicum annuum. Insects 2026, 17, 152. https://doi.org/10.3390/insects17020152
Chen X, Lin S, Linghu T, Yu Y, Li H, Chen Y, Wei H, Chen Y. Functional Characterization of CaSpr2 in Jasmonate-Dependent Induced Defense Against Western Flower Thrips in Capsicum annuum. Insects. 2026; 17(2):152. https://doi.org/10.3390/insects17020152
Chicago/Turabian StyleChen, Xi, Shuo Lin, Tingting Linghu, Yun Yu, Heng Li, Yixin Chen, Hui Wei, and Yong Chen. 2026. "Functional Characterization of CaSpr2 in Jasmonate-Dependent Induced Defense Against Western Flower Thrips in Capsicum annuum" Insects 17, no. 2: 152. https://doi.org/10.3390/insects17020152
APA StyleChen, X., Lin, S., Linghu, T., Yu, Y., Li, H., Chen, Y., Wei, H., & Chen, Y. (2026). Functional Characterization of CaSpr2 in Jasmonate-Dependent Induced Defense Against Western Flower Thrips in Capsicum annuum. Insects, 17(2), 152. https://doi.org/10.3390/insects17020152

