Enzymatic activity and development of Frankliniella occidentalis (Thysanoptera: Thripidae) in response to exogenous calcium treatments of kidney bean plants.

Frankliniella occidentalis Pergande (Thysanoptera: Thripidae) is an agricultural pest threatening various horticultural crops worldwide. Inducing plant resistance is an ecologically beneficial and potentially effective method for controlling F. occidentalis. As an essential nutrient element, exogenous calcium enhances plant-induced resistance. This study investigated the effects of CaCl2 on the secondary metabolites of kidney bean plants and detoxifying and digestive enzymes in F. occidentalis. We found that treatment of plants and treatment time and also the interactions of the 2 factors significantly affected secondary metabolites contents (tannin, flavonoids, total phenol, alkaloid, and lignin) of kidney bean leaves, which indicated that that the effect of treatment of plants on secondary metabolites varied with treatment time. Moreover, when thrips fed on CaCl2-treated plants, the activities of detoxifying enzymes, enzymes glutathione S-transferase and cytochrome P450 substantially increased compared to those in which thrips fed on control plants. However, the activity of carboxylesterase significantly decreased. The detoxifying enzyme genes CL992.contig6, CYP4PN1, and CYP4PJ2 were significantly upregulated at 24 and 48 h. The activities of digestive enzymes (α-amylase, chymotrypsin, and lipase) increased substantially in F. occidentalis. The digestive enzyme gene, FoAMY-1, was significantly upregulated at 24 and 48 h after treatment. The pupation rate and pupal weight of F. occidentalis were significantly reduced. The results indicated that exogenous CaCl2-induced metabolic changes in kidney bean plants and altered the enzymatic activity and development of F. occidentalis that fed upon them.

Frankliniella occidentalis pathogenic fungus Lecanicillium interacts with internal microbes and produces sublethal effects.

Frankliniella occidentalis (Thysanoptera: Thripidae) is a pest that feeds on various crops worldwide. A prior study identified Lecanicillium attenuatum and L. cauligalbarum as pathogens of F. occidentalis. Unfortunately, the potential of these two entomopathogenic fungi for the biocontrol of F. occidentalis has not been effectively evaluated. The internal microbes (endosymbionts and the gut microbiota) of insects, especially gut bacteria, are crucial in regulating the interactions between the host and intestinal pathogens. The role of thrips internal microbes in the infection of these two entomopathogenic fungi is also unknown. Therefore, biological control of thrips is immediately needed, and to accomplish that, an improved understanding of the internal microbes of thrips against Lecanicillium infection is essential. The virulence of the two pathogenic fungi against F. occidentalis increased with the conidia concentration. Overall, the LC50 of L. cauligalbarum was lower than that of L. attenuatum, and the pathogenicity degree was adult > pupa > nymphs. The activities of protective enzymes include superoxide dismutase (SOD), catalase (CAT), peroxidase (POD); detoxification enzymes include polyphenol oxidase (PPO), glutathione s-transferase (GSTs), and carboxylesterase (CarE); hormones include ecdysone and juvenile hormone; and the composition and proportion of microorganisms (fungi and bacteria) in F. occidentalis infected by L. cauligalbarum and L. attenuatum have changed significantly. According to the network correlation results, there was a considerable correlation among the internal microbes (including bacteria and fungi), enzyme activities, and hormones, which indicates that in addition to bacteria, internal fungi of F. occidentalis are also involved in the L. cauligalbarum and L. attenuatum infection process. In addition, the development time of the surviving F. occidentalis exposed to L. cauligalbarum or L. attenuatum was significantly shorter than that of the control group. Furthermore, the intrinsic rate of increase (rm), finite rate of increase (λ), net reproductive rate (R0), mean generation time (T), and gross reproductive rate (GRR) were significantly lower in the treatment groups than in the control group. L. attenuatum and L. cauligalbarum have biocontrol potential against F. occidentalis. In addition to bacteria, internal fungi of F. occidentalis are also involved in the infection process of insect pathogenic fungi. Disruption of the internal microbial balance results in discernible sublethal effects. Such prevention and control potential should not be ignored. These findings provide an improved understanding of physiological responses in thrips with altered immunity against entomopathogenic fungal infections, which can guide us toward the development of novel biocontrol strategies against thrips.

Multigenerational variation in the nutrients and digestion of western flower thrips (Frankliniella occidentalis) depends on the nutritive quality of different foods.

Western flower thrips (WFTs), Frankliniella occidentalis (Thysanoptera, Thripidae), is one of the most serious pests that attack rose flowers. Little is known about the effect of different parts of the rose flower on nutritional contents and digestive enzyme activities in thrips. This study assessed variations in the nutritional contents and digestive enzyme activities in the second-instar larvae and adults WFTs fed on 3 food types (rose petals, rose flowers, and honey solution + kidney bean pods) for multiple generations. The highest contents of soluble sugar (in 10% honey solution + kidney bean pods), amino acid (in rose flowers), and protein (in rose flowers) were observed, respectively. Soluble sugar and protein contents in the second-instar larvae and adults fed on rose petals decreased in the F1 generation but increased in the F2 generation and remained at higher levels until the F7 generation. Feeding of thrips with 3 food types increased the lipid content in the F1 generation, which peaked in the F2 generation and remained high until the F7 generation. In most cases, α-amylase and trypsin activities significantly decreased in the F1 generation after feeding on rose petals and then prominently increased in the F2 generation. In contrast, chymotrypsin activity remarkably increased and peaked in the F1 generation after second-instar larvae thrips fed on rose petals. There were correlations among the contents of 3 nutrient related positively with the activities of α-amylase and trypsin in WFTs second-instar larvae and adults, respectively. Overall, variations in the nutrient properties of the 3 food types caused changes in nutrient contents and digestive enzyme activities in thrips.

Effects of Different Parts of the Rose Flower on the Development, Fecundity, and Life Parameters of Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae).

Frankliniella occidentalis (Pergande) is an important horticultural pest that causes serious damage to rose plants, which is one of its preferred foods. In this study, rose petals, rose flowers, and 10% honey solution + kidney bean pods were chosen as foods to assess their influence on the growth, development and fecundity of F. occidentalis. The results showed that developmental time of immature F. occidentalis with the following trend: rose flowers <10% honey solution + kidney bean pods < rose petals < kidney bean pods. The longevities of both female and male adults were lowest when feeding on the rose petals and were highest when feeding on rose flowers. The fecundity was in the following order: rose flowers >10% honey solution + kidney bean pods > rose petals > kidney bean pods. The net reproductive rate (R0), intrinsic rate of increase (r), and finite rate of increase (λ) of F. occidentalis feeding on rose petals and kidney bean pods were lower than those feeding on rose flowers and 10% honey solution + kidney bean pods. The development, longevity, fecundity, and parameters have significantly changed since F1 generation after feeding with the three food types. The results indicated that different parts of rose flowers had a significant effect on the development of thrips, and nectar and pollen had a positive effect on thrips population increase and reproduction.

Exogenous Calcium Suppresses the Oviposition Choices of Frankliniella occidentalis (Thysanoptera: Thripidae) and Promotes the Attraction of Orius similis (Hemiptera: Anthocoridae) by Altering Volatile Blend Emissions in Kidney Bean Plants.

Frankliniella occidentalis is a destructive pest of horticultural plants, while Orius similis is a natural enemy of thrips. It has been demonstrated that exogenous calcium could induce plant defenses against herbivore attack. We examined whether CaCl2 supplementation altered the volatile emissions of kidney bean plants, which influence the oviposition preference of F. occidentalis. We also assessed the influence of volatile cues on O. similis. Using Y-tube olfactometer tests, we found that exogenous CaCl2 treatment inhibited the selectivity of F. occidentalis but attracted O. similis. In addition, CaCl2 treatment reduced the oviposition preference of F. occidentalis. Gas chromatography-mass spectrometry analyses revealed that CaCl2 treatment altered the number and relative abundance of the volatile compounds in kidney bean plants and that (E)-2-hexen-1-ol, 1-octen-3-ol, ß-lonone, and (E,E)-2,4-hexadienal might be potential olfactory cues. Furthermore, the results of the six-arm olfactometer test indicated that 1-octen-3-ol (10-2 µL/µL), ß-lonone (10-2 µL/µL), and (E,E)-2,4-hexadienal (10-3 µL/µL) repelled F. occidentalis but attracted O. similis. Overall, our results suggested that exogenous CaCl2 treatment induced defense responses in kidney bean plants, suggesting that CaCl2 supplementation may be a promising strategy to enhance the biological control of F. occidentalis.

Combined Jasmonic Acid and Ethylene Treatment Induces Resistance Effect in Faba Bean Plants Against Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae).

Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) is a serious invasive pest in China. In this study, we determined whether exogenous jasmonic acid (JA) and ethylene (ET) treatments could induce resistance against F. occidentalis in faba bean plants. First, we investigated the effects of different concentrations of JA or ET alone on F. occidentalis and then assessed the effects of optimal concentrations of JA and ET combined. Our results showed that the optimal concertation of JA was 2 mmol/L and ET was 0.5 mmol/L. JA + ET mixture showed the greatest inhibitory effect in terms of oviposition and feeding. JA with ET was found to induce changes in the activities of lipoxygenase (LOX), allene oxide synthase (AOS), polyphenol oxidase (PPO), 1-aminocyclopropane 1-carboxylic acid synthase (ACS), and trypsin inhibitor (TI). This treatment also activated or inhibited the relative expression levels of LOX1, ACO2, ACS2, and AP2/ERF. Treatment of faba bean plants with JA and ET significantly prolonged F. occidentalis development and adult preoviposition period, significantly reduced per-female oviposition, and altered male longevity and offspring demographic parameters. These results indicate that JA with ET can induce defenses against the growth and development of F. occidentalis in faba bean plants.

Copper Chaperone for Superoxide Dismutase FoCCS1 in Frankliniella occidentalis May Be Associated with Feeding Adaptation after Host Shifting.

Western flower thrips (Frankliniella occidentalis) pose a serious threat to the global vegetable and flower crop production. The regulatory mechanism for superoxide dismutase (SOD) in the feeding adaptation of F. occidentalis after host shifting remains unclear. In this study, the copper chaperone for SOD (CCS) and manganese SOD (MnSOD) genes in F. occidentalis were cloned, and their expression levels at different developmental stages was determined. The mRNA expression of FoCCS1 and FoMnSOD2 in F. occidentalis second-instar larvae and adult females of F1, F2, and F3 generations was analyzed after shifting the thrips to kidney bean and broad bean plants, respectively. The F2 and F3 second-instar larvae and F2 adult females showed significantly upregulated FoCCS1 mRNA expression after shifting to kidney bean plants. The F1 second-instar larvae and F2 adult females showed significantly upregulated FoCCS1 mRNA expression after shifting to broad bean plants. The RNA interference significantly downregulated the FoCCS1 mRNA expression levels and adult females showed significantly inhibited SOD activity after shifting to kidney bean and broad bean plants. F. occidentalis adult females subjected to RNA interference and released on kidney bean and broad bean leaves for rearing, respectively, significantly reduced the survival rate and fecundity. These findings suggest that FoCCS1 plays an active role in regulating the feeding adaptation ability of F. occidentalis after host shifting.

Estimated Divergence Times of Lecanicillium in the Family Cordycipitaceae Provide Insights Into the Attribution of Lecanicillium.

Background: The genus Lecanicillium W.Gams & Zare is a recognized insect pathogen but members of the genus have been found parasitizing various hosts including arthropods, nematodes, plants, and fungi. The new classification system for fungi proposed to reject Lecanicillium and transfer some of the species to the genus Akanthomyces. However, the attribution problem of most species in the original genus Lecanicillium remains unsolved. The current study aimed to improve understanding of the pivotal internal phylogeny in Lecanicillium by estimating the divergence times of Lecanicillium to provide additional insights into the status of this genus within the family Cordycipitaceae. Results: Dating analyses support the supposition that the ancestor of Lecanicillium was in the Cretaceous period (84.36 Mya, 95% HPD: 72.12-94.74 Mya). After originating from a common ancestor, eight clades of Lecanicillium were derived and evolved independently in parallel with other genera of Cordycipitaceae. Based on the clear divergence age estimates, Lecanicillium clade 8 originated earlier as an independent group in the Cretaceous period (75.61 Mya, 95% HPD: 63.31-87.54 Mya), while Lecanicillium clades 1-7 originated later as an independent group in the boundary of the Cretaceous and Paleogene periods (64.66 Mya, 95% HPD: 52.75-76.74 Mya). Lecanicillium huhutii formed an independent branch in a polytomy together with a clade containing Lecanicillium tenuipes (BI posterior probabilities 1, ML bootstrap 100%). Conclusion: The pivotal internal phylogeny, origin, and evolutionary history of Lecanicillium in the family Cordycipitaceae were investigated. Phylogenetic and morphological analyses indicated that there are eight representative clades (four representative branches of evolutionary history), including clade 1 (members have a relatively uniform sporulation structure comprising globose heads with a higher number of conidia), clade 8 (including all members of Gamszarea), clades 2-5 (the differences of the divergence time estimations were smaller compared with other clades), and clade 6-7 (members are close to Gibellula, Hevansia, and Ascopolyporus). Based on the above findings, a novel spider-pathogenic fungus, Lecanicillium huhutii, is described. All other species in Lecanicillium clade 1 (Lecanicillium araneogemum, L. nodulosum, L. pissodis, and L. uredinophilum) should be transferred to the genus Akanthomyces. Furthermore, the monotypic genus Parengyodontium should be merged with the genus Gamszarea. More novel species need to be discovered to thoroughly resolve the attribution problem of Lecanicillium. Finally, no major lineages of Lecanicillium were correlated with the nearby Cretaceous-Tertiary extinction event, indicating that the diversity of Lecanicillium is more likely to be caused by long-term environmental adaptation and coevolution with insects rather than by dramatic extinction events.

Life Table and Preference Choice of Frankliniella occidentalis (Thysanoptera: Thripidae) for Kidney Bean Plants Treated by Exogenous Calcium.

Exogenous calcium (Ca) has been used to induce host plant resistance in response to abiotic and biotic stresses, including from thrips attack. The aim of this study was to determine whether exogenously applied Ca affects the performance of Frankliniella occidentalis. We assessed the development time, total longevity, reproduction, and population parameters of F. occidentalis, and its preference choice on Ca-treated or untreated control kidney bean plants under laboratory conditions. The results showed that F. occidentalis fed on Ca-treated leaves had a longer developmental time but lower longevity (female and male) and fecundity than F. occidentalis fed on control leaves. Population parameters, including the intrinsic rate of increase (r), finite rate of increase (λ), and net reproductive rate (R0), were all found higher in control leaves than in Ca-treated leaves, and the mean generation time (T) was shorter. In preference choices, the number of thrips on control plants was higher than the number of thrips on Ca-treated kidney bean plants. Overall, our results indicated that exogenous Ca pretreatment on kidney bean plants affected the life history and preference choice of F. occidentalis, suggesting Ca might be used as a promising elicitor of inducible plant defense against thrips.

Establishment of a Faba Bean Banker Plant System with Predator Orius strigicollis for the Control of Thrips Dendrothrips minowai on Tea Plants under Laboratory Conditions.

The stick tea thrip Dendrothrips minowai (Priesner) (Thysanoptera: Thripidae) is a destructive pest in tea plantations in south and southwest China. To control this pest, a non-crop banker plant system was developed using a polyphagous predator Orius strigicollis (Poppius) (Heteroptera: Anthocoridae) with the black bean aphid Aphis fabae (Scopoli) (Hemiptera: Aphididae) as an alternative prey and the faba bean Vicia faba as the banker plant to support the predator in targeting the pest. The fitness of A. fabae on tea plants and faba bean was evaluated to determine its host specificity. Moreover, the control efficacy of the banker plant system on D. minowai on tea plants was tested in the laboratory and compared with that of direct release of O. strigicollis. The experiments showed that faba bean was an excellent non-crop host for A. fabae because, while the aphid population increased quickly on faba bean, it could only survive for up to 9 days on tea plants. Compared with direct release of O. strigicollis, lower densities of pest were observed when introducing the banker plant system. Our results indicate that this banker plant system has the potential to be implemented in the field to improve the control of the pest thrips.

Orius similis (Hemiptera: Anthocoridae): A Promising Candidate Predator of Spodoptera frugiperda (Lepidoptera: Noctuidae).

The bug Orius similis Zheng is a native generalist predator of insect pests in southern China. The fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith), is a notorious defoliator that is now an economically important insect pest throughout China. To investigate the ability of O. similis to control FAW, we evaluated the predatory capacity, behavior, and functional response of O. similis with respect to FAW and their olfactory response. Both females and males successfully preyed on FAW eggs and first-instar larvae but not on second-instar or older larvae. Adult O. similis only attacked and killed one egg or one larva at a time before sucking the prey, and similar predatory behavior was also observed with regard to FAW egg masses. Both female and male O. similis exhibited type II functional response when preying on FAW eggs and first-instar larvae. Maximum estimated prey consumed per day was 23.7 eggs and 26.2 larvae for adult females and 22.5 eggs and 19.6 larvae for adult males. Moreover, in a Y-tube olfactometer experiment, both female and male O. similis exhibited a significant preference for maize (Zea mays L.) seedlings damaged by FAW over clean air, healthy seedlings, FAW feces, and FAW, suggesting that seedlings damaged by FAW may releases specific volatiles attracting the predator. Collectively, the results of the study suggest that O. similis is a promising candidate for the biological control of FAW eggs and first-instar larvae, particularly given its attraction to FAW-damaged leaves, which may enable it to locate the target prey rapidly.

Response of Protective Enzymes in Western Flower Thrips (Thysanoptera: Thripidae) to Two Leguminous Plants.

The western flower thrips, Frankliniella occidentalis, is a major invasive pest of commercially important crops worldwide. We compared the activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) and the expressions of two putative SOD and two putative POD sequences in second instar larvae and adults after three generations of adaptation to kidney bean and broad bean plants. The results showed that the SOD, POD, and CAT activities in adults were significantly higher than those in the second instar larvae. The SOD activities were significantly higher in both the second instar larvae and the adults fed on kidney bean (Phaseolus vulgaris) plants versus broad bean (Vicia faba) plants, whereas the POD and CAT activities showed the opposite trend. The gene expression data showed that the FoPOD-2 expression levels were lower in the second instar larvae after three generations of feeding on broad bean plants versus kidney bean plants. The expression levels of FoSOD-1 and FoSOD-2, and FoPOD-1 under broad bean plant treatment were higher than those under kidney bean plant treatment. Additionally, gene expression fluctuated among the different generations. Our results indicated that western flower thrips demonstrated plasticity in gene expression and activity of protective enzymes, which is related to their adaptability to the host plants. Western flower thrips can change the expression of protective enzyme genes and enzyme activity in vivo to better adapt to kidney bean and broad bean plants.

Lecanicilliumcauligalbarum sp. nov. (Cordycipitaceae, Hypocreales), a novel fungus isolated from a stemborer in the Yao Ren National Forest Mountain Park, Guizhou.

A new species of entomopathogenic fungi, Lecanicilliumcauligalbarum, was discovered from a survey of invertebrate-associated fungi in the Yao Ren National Forest Mountain Park in China. The synnemata of this species emerged from the corpse of a stemborer (Lepidoptera), which was hidden amongst pieces of wood on the forest floor. It differs from morphologically similar Lecanicillium species mainly in its short conidiogenous cells and ellipsoid to ovoid and aseptate conidia. Phylogenetic analysis of a combined data set comprising ITS, SSU, LSU, TEF, RPB1 and RPB2 sequence data supported the inclusion of L.cauligalbarum in the Lecanicillium genus and its recognition as a distinct species.

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To evaluate the sublethal effects of spinetoram and azadirachtin on western flower thrips, Frankliniella occidentalis, leaf dipping method was used to determine their sublethal concentrations (LC25) on the 2nd instar nymph, and their influences on development and reproduction of F. occidentalis were studied. The results showed exposure of sublethal concentrations of spinetoram and azadirachtin to F. accidentalis had different degrees of effects on this insect pest. Under bisexual reproduction, the LC25 spinetoram had no significant influences on pre-oviposition period, female adult longevity and fecundity, but male adult longevity was significantly shorter than the control. The LC25 azadirachtin significantly reduced fecundity and prolonged pre-oviposition period. Under parthenogenesis, the LC25 spinetoram and azadirachtin extended the pre-oviposition duration, whereas the LC25 azadirachtin shortened the female adult longevity and significantly decreased fecundity. The LC25 spinetoram and azadirachtin had different influences on developmental duration of each stage of next generation. The immature stage in treatment group of the LC25 spinetoram was shorter than that in treatment group of the LC25 azadirachtin, under bisexual reproduction or parthenogenesis. Intrinsic rate of increase (rm) and finite rate of increase (λ) of population treated by the LC25 spinetoram were higher than those of the control, whereas the rm, R0, and λ of population treated by the LC25 azadirachtin were lower than those of the control. The findings indicated that the effects of the LC25 spinetoram and azadirachtin on the development and reproduction of F. accidentalis were different. The LC25 spinetoram had certain stimulating effect, whereas the LC25 azadirach-tinon had significant inhibitory effect. Two biopesticides' influences were related with the reproductive patterns of F. accidentalis.