Colorado potato beetles (leptinotarsa decemlineata) adapt to proteinase inhibitors induced in potato leaves by methyl jasmonate

doi:10.1016/0022-1910(95)00073-4

Journal of Insect Physiology

Volume 41, Issue 12, December 1995, Pages 1071-1078

https://doi.org/10.1016/0022-1910(95)00073-4 Get rights and content

Abstract

Potato plants were treated with gaseous methyl jasmonate (MJ) to obtain leaves with high induced levels of cysteine and aspartic proteinase inhibitors. Induced papain inhibitor activity was estimated at 4% of total protein. Other conditions produced leaves with low and moderate levels of this inhibitor. Development of Colorado potato beetle larvae was similar when they were reared on leaves containing low, moderate and high levels of papain inhibitor. Nevertheless, general proteinase activity was significantly reduced (42%) in insects reared on the high inhibitor diet, while proteinase activity that was insensitive to induced inhibitors in juice from MJ-treated leaves had increased two-fold. Activities towards the specific cysteine proteinase substrate p-Glu-Phe-Leu-pNA were the same in guts from insects reared on the three leaf types. However, juice from MJ-treated leaves inhibited as much as 67% of this activity in guts of insects reared on the low inhibitor diet compared to only 27% of the activity in gut extracts from insects reared on MJ-treated leaves, indicating a 2.5-fold induction of cysteine proteinase activity insensitive to potato proteinase inhibitors. None of the activities towards another specific cysteine proteinase substrate l-Arg-pNA were sensitive to inhibitors from MJ-treated leaves, but guts of insects fed these leaves had a 3.5-fold induction of this proteinase activity compared to those reared on plants containing low papain inhibitor levels. These data suggest that Colorado potato beetle larvae compensated for inhibited gut proteolytic activity during chronic intake of papain inhibitors by synthesizing insensitive proteinase(s).

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Protease inhibitors (PIs) have been traditionally recognized by their potential biomedical application in events with exacerbation of endogenous proteases activity. Plant PIs have gained interest as naturally occurring molecules, which usually show lower environmental impact residual toxicity than synthetic compounds. In this work, we isolated, cloned, expressed and purified a novel trypsin inhibitor from S. tuberosum subsp. andigenum var. overa, named oPTI. A significant over-expression of the oPTI coding gene after 48 h exposure of methyl jasmonate compared to the gene of reference. This inhibitor showed a molecular mass of 12 kDa and a Ki of 7.3 × 10⁻⁷ M. Finally, we evaluated the antimicrobial activity of oPTI against different pathogenic microorganisms. The oPTI demonstrated inhibitory effect on the growth of Acinetobacter baumannii S-1, Acinetobacter baumannii R, Acinetobacter calcoaceticus R, Acinetobacter calcoaceticus S, Bacillus stearothermophilus, Escherichia coli, Pseudomonas aeruginosa, Salmonella braenderup, Salmonella enteritidis, Salmonella typhimurium and Yersinia enterocolitica strains. This study represents the first report for the antimicrobial activity of a plant PI over a wide range of microorganisms. Our studies reinforce the importance of natural PIs as promising molecules for their potential application in the biomedical field and/or in the food industry as natural food preservatives.
Insect predator odors protect herbivore from fungal infection
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One way that researchers manipulate host plant defenses is to treat foliage with solutions of signaling molecules, like jasmonic acid or salicylic acid. Such treatments have been shown to induce the production of defensive compounds like protease inhibitors and polyphenoloxidase in solanaceous plants including potato (Bolter and Jongsma 1995, Rivard et al. 2004). Colorado potato beetles are a devastating pest of solanaceous crops like potato, Solanum tuberosum L., due in part to their ability to rapidly develop resistance to synthetic chemical insecticides (Whalon et al. 2013).
Lethal predators and pathogens can reduce the size of pest populations and help regulate crop-damaging herbivores. Additionally, predators like the spined soldier bug (Podisus maculiventris) can elicit non-consumptive effects in herbivores such as the Colorado potato beetle (Leptinotarsa decemlineata), causing changes in prey behavior, physiology and life history. While it is known that lethal predators and pathogens interact, there is little known about the interaction of predator-induced non-consumptive effects and pathogens on their shared hosts. Using the spined soldier bug, which is known to elicit non-consumptive effects on potato beetles (e.g. reduced feeding), we studied the interaction of predator-induced non-consumptive effects and the fungal pathogen Beauveria bassiana on the survival of Colorado potato beetle larvae. In laboratory microcosms we found a 16–50% increase in survival of fungus-treated beetles held with non-lethal male stink bugs. We also studied the predator-fungus interaction on jasmonic acid-treated plants to assess whether elevated levels of host plant defenses made larvae more susceptible to fungal infection. Beetles’ susceptibility to fungal infection did not change when provided jasmonic-acid treated foliage as food. Finally, we determined that two volatile compounds (E-2-hexenal and alpha-terpineol), produced as part of the bugs’ aggregation pheromone, reduced conidial germination, resulting in higher levels of beetle survival. Our findings suggest that stink bug odors protect prey against fungal pathogens.
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Colorado potato beetle (CPB) is one of the most devastating invasive insects and it is native to North America. It feeds on several wild species of the genus Solamum, such as S. elaeagnifolium and S. rostratum Dunal, and is one of the major pests of potato and eggplant. Beginning in the early 19th century, CPB has rapidly spread across North America, Europe, and Central Asia. CPB was first reported to invade Xinjiang of China in 1993 and it was effectively controlled in Mori County. Since 2013, CPB has also been found in Jilin and Heilongjiang in Northeast China, and it likely migrated to these provinces from Russia. Thus, China has become the frontier for the global CPB spread, and risk management and monitoring systems for this pest are urgently needed. Here, we summarize pest management methods that are used in areas at the frontier of the CPB invasion, and put forward frameworks for further preventing and controlling of the spread of CPB. The management methods for CPB can also serve as an example for the control of invasive species mitigation in frontier areas.
Population-associated heterogeneity of the digestive Cys protease complement in Colorado potato beetle, Leptinotarsa decemlineata
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Citation Excerpt :
An interesting case is L. decemlineata, which has become somewhat of a model species for the study of herbivorous insect proteases and protease–inhibitor interactions in plant/insect systems (Cingel et al., 2016; Oppert et al., 2014; Smid et al., 2013; Vorster et al., 2015). A wealth of literature has described basic midgut protease profiles (Brunelle et al., 1999; Goulet et al., 2008; Gruden et al., 1998, 2003, 2004; Michaud et al., 1993; Michaud et al., 1995, 1996; Novillo et al., 1997; Sainsbury et al., 2012b; Srp et al., 2016; Thie and Houseman, 1990; Visal et al., 1998; Visal-Shah et al., 2001; Vorster et al., 2015) and protease-related compensatory processes in response to dietary challenges (Bolter and Jongsma, 1995; Bolter and Latoszek-Green, 1997; Brunelle et al., 2004; Cloutier et al., 1999, 2000; Gruden et al., 2003, 2004; Overney et al., 1997; Rivard et al., 2004; Smid et al., 2013, 2015) in L. decemlineata. By contrast, no article yet has assessed midgut complement variability in different populations of this species and the eventual consequences of such variability on the potency of recombinant inhibitor candidates to protect potato in different agricultural regions.
Herbivorous insects use complex protease complements to process plant proteins, useful to adjust their digestive functions to the plant diet and to elude the antidigestive effects of dietary protease inhibitors. We here assessed whether basic profiles and diet-related adjustments of the midgut protease complement may vary among populations of the insect herbivore Colorado potato beetle (Leptinotarsa decemlineata). Two laboratory colonies of this insect were used as models, derived from insect samples collected in potato fields ∼1200 km distant from each other in North America. Synchronized 4th-instar larvae reared on potato were kept on this plant, or switched to tomato or eggplant, to compare their midgut cathepsin activities and content of intestain Cys proteases under different diet regimes. Cathepsin D activity, cathepsin L activity, cathepsin B activity and total intestain content shortly after larval molting on potato leaves were about two times lower in one population compared to the other. By comparison, cathepsin D activity, cathepsin B activity, total intestain content and relative abundance of the most prominent intestain families were similar in the two populations after three days regardless of the plant diet, unlike cathepsin L activity and less prominent intestain families showing population-associated variability. Variation in Cys protease profiles translated into the differential efficiency of a Cys protease inhibitor, tomato cystatin SlCYS8, to inhibit cathepsin L activity in midgut extracts of the two insect groups. Despite quantitative differences, SlCYS8 single variants engineered to strongly inhibit Cys proteases showed improved potency against cathepsin L activity of either population. These data suggest the feasibility of designing cystatins to control L. decemlineata that are effective against different populations of this insect. They underline, on the other hand, the practical relevance of considering natural variability of the protease complement among L. decemlineata target populations, eventually determinant in the success or failure of cystatin-based control strategies on a large-scale basis.

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Colorado potato beetles (leptinotarsa decemlineata) adapt to proteinase inhibitors induced in potato leaves by methyl jasmonate

Abstract

Analyt Biochem.

J. Insect Physiol.

J. Insect Physiol.

Prot. Express. Purif.

Analyt Biochem.

J. Biol. Chem.

J. Insect Physiol.

J. Insect Physiol.

FEBS Lett

Comp. Biochem. Physiol.

J. Insect Physiol.

Insect Biochem. Molec. Biol.

Insect Biochem.

Biochem. Biophys. Acta

Simultaneous formation of chymopapain inhibitor and cubical crystals in tomato leaves

Can. J. Bot.

Effect of wounding and jasmonates on the physico-chemical properties and flea beetles defence response of canola seedlings, Brassica napus. L

Can J. Plant Sci.

Effect of induced resistance mechanisms in potato and tomato plants on the Colorado potato beetle

Methyl jasmonate induces papain inhibitor(s) in tomato leaves

Plant Physiol.