Feeding of Whitefly on Tobacco Decreases Aphid Performance via Increased Salicylate Signaling

Background

The feeding of Bemisia tabaci nymphs trigger the SA pathway in some plant species. A previous study showed that B. tabaci nymphs induced defense against aphids (Myzus persicae) in tobacco. However, the mechanism underlying this defense response is not well understood.

Methodology/Principal Findings

Here, the effect of activating the SA signaling pathway in tobacco plants through B. tabaci nymph infestation on subsequent M. persicae colonization is investigated. Performance assays showed that B. tabaci nymphs pre-infestation significantly reduced M. persicae survival and fecundity systemically in wild-type (WT) but not salicylate-deficient (NahG) plants compared with respective control. However, pre-infestation had no obvious local effects on subsequent M. persicae in either WT or NahG tobacco. SA quantification results indicated that the highest accumulation of SA was induced by B. tabaci nymphs in WT plants after 15 days of infestation. These levels were 8.45- and 6.14-fold higher in the local and systemic leaves, respectively, than in controls. Meanwhile, no significant changes of SA levels were detected in NahG plants. Further, biochemical analysis of defense enzymes polyphenol oxidase (PPO), peroxidase (POD), β-1,3-glucanase, and chitinase demonstrated that B. tabaci nymph infestation increased these enzymes’ activity locally and systemically in WT plants, and there was more chitinase and β-1, 3-glucanase activity systemically than locally, which was opposite to the changing trends of PPO. However, B. tabaci nymph infestation caused no obvious increase in enzyme activity in any NahG plants except POD.

Conclusions/Significance

In conclusion, these results underscore the important role that induction of the SA signaling pathway by B. tabaci nymphs plays in defeating aphids. It also indicates that the activity of β-1, 3-glucanase and chitinase may be positively correlated with resistance to aphids.     

Root and shoot jasmonic acid induced plants differently affect the performance of Bemisia tabaci and its parasitoid Encarsia formosa

Plants employ both direct and indirect defenses to protect themselves from attacks by herbivores and pathogens. To date most aboveground and belowground interaction studies have focused on interactions between plants and leaf-chewing herbivores, while the plant defence on the performance of phloem-feeding insects, induced by above- and belowground interaction, has been less explored. Here, jasmonic acid (JA) was used to mimic herbivore-induced responses in Chinese broccoli (Brassica oleracea var. alboglabra) roots (RJA) and shoots (SJA). The effects of JA-induced plant defenses on the performance of the phloem-feeding whitefly, Bemisia tabaci, and its aphelinid parasitoid Encarsia formosa were investigated. The results indicated that SJA induction has a much larger negative effect on B. tabaci than RJA: nymphs develop slower and have a lower survivorship. Also, females live shorter and have a lower fecundity on SJA plants compared to those on RJA and untreated control (CON) plants. The intrinsic rate of increase (r_m) of B. tabaci on SJA plants was 0.089, which was significantly lower than those on CON and RJA plants (0.115 and 0.104, respectively). The parasitoid E. formosa, on the other hand, shows a significantly faster development when parasitizing whitefly hosts feeding on SJA plants, whereas parasitism rate, longevity and fecundity were similar to those on RJA and CON plants. The current study reveals that plants induced with exogenous JA vary in both their resistance to whitefly and suitability for parasitoids, depending on the organ to which the JA was applied. Root and shoot JA applications also have contrasting effects on the phloem-feeding insect B. tabaci and its parasitoid, that is, SJA induction leads to more negative effects on whitefly performance than RJA induction, but its parasitoid performs better on hosts reared on SJA plants. These results show that top-down and bottom-up processes governing herbivore populations on Chinese broccoli are working in concert to increase plant resistance when plants are induced by SJA application.     

Plant Virus Differentially Alters the Plant's Defense Response to Its Closely Related Vectors

Background

The whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae), is one of the most widely distributed agricultural pests. In recent years, B. tabaci Q has invaded China, and Q has displaced B in many areas now. In a number of regions of the world, invasion by B and/or Q has been followed by outbreaks of tomato yellow leaf curl virus (TYLCV). Our previous study showed TYLCV directly and indirectly modified the feeding behavior of B. tabaci in favor of Q rather than B.

Methodology/Principal Findings

In this study, we quantified the salicylic acid (SA) titers and relative gene expression of SA in tomato leaves that were infested with viruliferous or non-viruliferous B and Q. We also measured the impacts of exogenous SA on the performance of B and Q, including the effects on ovary development. SA titer was always higher in leaves that were infested with viruliferous B than with viruliferous Q, whereas the SA titer did not differ between leaves infested with non-viruliferous B and Q. The relative gene expression of SA signaling was increased by feeding of viruliferous B but was not increased by feeding of viruliferous Q. The life history traits of B and Q were adversely affected on SA-treated plants. On SA-treated plants, both B and Q had lower fecundity, shorter longevity, longer developmental time and lower survival rate than on untreated plants. Compared with whiteflies feeding on control plants, those feeding on SA-treated plants had fewer oocytes and slower ovary development. On SA-treated plants, viruliferous B had fewer oocytes than viruliferous Q.

Conclusions/Significance

These results indicate that TYLCV tends to induce SA-regulated plant defense against B but SA-regulated plant defense against Q was reduced. In other words, Q may have a mutualistic relationship with TYLCV that results in the reduction of the plant''s defense response.     

Allergenicity assessment of Allium sativum leaf agglutinin, a potential candidate protein for developing sap sucking insect resistant food crops

Background

Mannose-binding Allium sativum leaf agglutinin (ASAL) is highly antinutritional and toxic to various phloem-feeding hemipteran insects. ASAL has been expressed in a number of agriculturally important crops to develop resistance against those insects. Awareness of the safety aspect of ASAL is absolutely essential for developing ASAL transgenic plants.

Methodology/Principal Findings

Following the guidelines framed by the Food and Agriculture Organization/World Health Organization, the source of the gene, its sequence homology with potent allergens, clinical tests on mammalian systems, and the pepsin resistance and thermostability of the protein were considered to address the issue. No significant homology to the ASAL sequence was detected when compared to known allergenic proteins. The ELISA of blood sera collected from known allergy patients also failed to show significant evidence of cross-reactivity. In vitro and in vivo assays both indicated the digestibility of ASAL in the presence of pepsin in a minimum time period.

Conclusions/Significance

With these experiments, we concluded that ASAL does not possess any apparent features of an allergen. This is the first report regarding the monitoring of the allergenicity of any mannose-binding monocot lectin having insecticidal efficacy against hemipteran insects.     

Defense suppression benefits herbivores that have a monopoly on their feeding site but can backfire within natural communities

Background

Plants have inducible defenses to combat attacking organisms. Hence, some herbivores have adapted to suppress these defenses. Suppression of plant defenses has been shown to benefit herbivores by boosting their growth and reproductive performance.

Results

We observed in field-grown tomatoes that spider mites (Tetranychus urticae) establish larger colonies on plants already infested with the tomato russet mite (Aculops lycopersici). Using laboratory assays, we observed that spider mites have a much higher reproductive performance on russet mite-infested plants, similar to their performance on the jasmonic acid (JA)-biosynthesis mutant def-1. Hence, we tested if russet mites suppress JA-responses thereby facilitating spider mites. We found that russet mites manipulate defenses: they induce those mediated by salicylic acid (SA) but suppress those mediated by JA which would otherwise hinder growth. This suppression of JA-defenses occurs downstream of JA-accumulation and is independent from its natural antagonist SA. In contrast, spider mites induced both JA- and SA-responses while plants infested with the two mite species together display strongly reduced JA-responses, yet a doubled SA-response. The spider mite-induced JA-response in the presence of russet mites was restored on transgenic tomatoes unable to accumulate SA (nahG), but russet mites alone still did not induce JA-responses on nahG plants. Thus, indirect facilitation of spider mites by russet mites depends on the antagonistic action of SA on JA while suppression of JA-defenses by russet mites does not. Furthermore, russet mite-induced SA-responses inhibited secondary infection by Pseudomonas syringae (Pst) while not affecting the mite itself. Finally, while facilitating spider mites, russet mites experience reduced population growth.

Conclusions

Our results show that the benefits of suppressing plant defenses may diminish within communities with natural competitors. We show that suppression of defenses via the JA-SA antagonism can be a consequence, rather than the cause, of a primary suppression event and that its overall effect is determined by the presence of competing herbivores and the distinct palette of defenses these induce. Thus, whether or not host-defense manipulation improves an herbivore’s fitness depends on interactions with other herbivores via induced-host defenses, implicating bidirectional causation of community structure of herbivores sharing a plant.

     

The role of jasmonic acid and ethylene crosstalk in direct defense of Nicotiana attenuata plants against chewing herbivores

We examined performance of herbivores on plants lacking either jasmonate (JA, asLOX3) or ethylene (ET, mETR1) signaling or both (mETR1asLOX3). Plant defenses against Manduca sexta caterpillars were strongly impaired in JA-deficient asLOX3 plants; however, making asLOX3 plants ethylene insensitive did not further increase the performance of the larvae on a mETR1asLOX3 genetic cross. This result demonstrates the dominant role of JA over ET in the regulation of plant defenses against herbivores. However, ET-insensitivity combined with otherwise normal levels of JA in mETR1 plants promoted faster caterpillar growth, which correlated with reduced accumulation of the alkaloidal direct defense nicotine in mETR1 compared to WT plants. Our data points to an important accessory function of ET in the activation of JA-regulated plant defenses against herbivores at the level of alkaloid biosynthesis in the roots and/or accumulation in the leaves.Key words: herbivory, jasmonic acid and ethylene crosstalk, Nicotiana attenuata, nicotine, trypsin proteinase inhibitors (TPIs)     

Towards a reporter system to identify regulators of cross-talk between salicylate and jasmonate signaling pathways in Arabidopsis

The plant signaling hormones salicylic acid (SA) and jasmonic acid (JA) are regulators of inducible defenses that are activated upon pathogen or insect attack. Cross-talk between SA- and JA-dependent signaling pathways allows a plant to finely tune its response to the attacker encountered. In Arabidopsis, pharmacological experiments revealed that SA exerts a strong antagonistic effect on JA-responsive genes, such as PDF1.2, indicating that the SA pathway can be prioritized over the JA pathway. SA-mediated suppression of the JA-responsive PDF1.2 promoter was exploited for setting up a genetic screen aiming at the isolation of signal transduction mutants that are impaired in this cross-talk mechanism. The PDF1.2 promoter was fused to the herbicide resistance gene BAR to allow for life/death screening of a population of mutagenized transgenic plants. Non-mutant plants should survive herbicide treatment when methyl jasmonate (MeJA) is applied, but suppression of the JA response by SA should be lethal in combination with the herbicide. Conversely, crucial SA/JA cross-talk mutants should survive the combination treatment. SA effectively suppressed the expression of the PDF1.2::BAR transgene. However, suppression of the BAR gene did not result in suppression of herbicide resistance. Hence, a screening method based on quantitative differences in the expression of a reporter gene may be better suited to identify SA/JA cross-talk mutants. Here, we demonstrate that the PDF1.2::GUS reporter will be excellently suited in this respect.Key words: plant defense, salicylic acid, jasmonic acid, cross-talk, mutant screen, Arabidopsis     

Herbivore and phytohormone induced defensive response in kale against cabbage butterfly, Pieris brassicae Linn

The constitutive and induced resistance were studied in two varieties (Khanyari and Kawdari) of kale, Brassica oleracea var. acephala in response to cabbage butterfly, Pieris brassicae infestation and exogenous application of jasmonic acid (JA) and salicylic acid (SA). Phenols, condensed tannins, flavonoids and proteins were measured at six days after JA (1?mM) and SA (1?mM) application and/or insect infestation. Plant damage and larval weights were also recorded. Khanyari variety showed highest amounts of phenols (208.23?μg/g FW), condensed tannin (347.76?μg/g FW), flavonoids (175.61?μg/g FW) and proteins (0.71?mg/g FW) in plants pre-treated with JA and infested with insects. The PAL activity was high in response to JA application followed by insect infestation. Insects reared on Khanyari and Kawdari plants pre-treated with JA prior showed significantly reduced larval weights (97.88 and 102.46?mg, respectively). Damage was low in plants pre-treated with JA in Khanyari at 3, 6 and 9?days after treatment (10.52%, 8.52%, and 5.30%, respectively). Thus, JA can play an important role in plant defense in kale against P. brassicae.     

Plant-Generated Artificial Small RNAs Mediated Aphid Resistance

Background

RNA silencing is an important mechanism for regulation of endogenous gene expression and defense against genomic intruders in plants. This natural defense system was adopted to generate virus-resistant plants even before the mechanism of RNA silencing was unveiled. With the clarification of that mechanism, transgenic antiviral plants were developed that expressed artificial virus-specific hairpin RNAs (hpRNAs) or microRNAs (amiRNAs) in host plants. Previous works also showed that plant-mediated RNA silencing technology could be a practical method for constructing insect-resistant plants by expressing hpRNAs targeting essential genes of insects.

Methodology/Principal findings

In this study, we chose aphid Myzus persicae of order Hemiptera as a target insect. To screen for aphid genes vulnerable to attack by plant-mediated RNA silencing to establish plant aphid resistance, we selected nine genes of M. persicae as silencing targets, and constructed their hpRNA-expressing vectors. For the acetylcholinesterase 2 coding gene (MpAChE2), two amiRNA-expressing vectors were also constructed. The vectors were transformed into tobacco plants (Nicotiana tabacum cv. Xanti). Insect challenge assays showed that most of the transgenic plants gained aphid resistance, among which those expressing hpRNAs targeting V-type proton ATPase subunit E-like (V-ATPaseE) or tubulin folding cofactor D (TBCD) genes displayed stronger aphicidal activity. The transgenic plants expressing amiRNAs targeting two different sites in the MpAChE2 gene exhibited better aphid resistance than the plants expressing MpAChE2-specific hpRNA.

Conclusions/Significance

Our results indicated that plant-mediated insect-RNA silencing might be an effective way to develop plants resistant to insects with piercing-sucking mouthparts, and both the selection of vulnerable target genes and the biogenetic type of the small RNAs were crucial for the effectiveness of aphid control. The expression of insect-specific amiRNA is a promising and preferable approach to engineer plants resistant to aphids and, possibly, to other plant-infesting insects.     

Root nematode infection enhances leaf defense against whitefly in tomato

The foliar response to different herbivores sharing the same hosts is an important topic for the study of plant-insect interactions. Plants evolve local and systemic resistant strategies to cope with herbivores. Many researchers have characterized the mechanisms of leaf responses to insect infestation; however, the fact that roots serve as systemic resistance modulators to leaf herbivores has been widely ignored. Here, we report that tomato (Solanum lycopersicum) plants infected with southern root-knot nematodes (Meloidogyne incognita)—which feed on the roots to form nodules—enhanced leaf defenses against aboveground attackers, specifically, the whitefly (Bemisia tabaci). Our results show that nematode infection reduced the whitefly population abundance because of conferring a stronger SA-dependent defense pathway against whitefly than in tomato plants without nematode infection. Meanwhile, nematode-infected tomato plant also activated the foliar JA-dependent defense pathway at 4 h after whitefly infestation. However, the foliar JA-dependent defense under whitefly infestation alone was suppressed, with the JA content being nearly 30 % lower than that in tomato plants co-infected with nematodes and whiteflies. Furthermore, nematode infection significantly decreased the plant nitrogen concentration in leaves and roots. As a result, nematode infection reduced the number of whiteflies by enhancing foliar SA-dependent defense, activating JA-dependent defense and decreasing nitrogen nutrition. Our results suggest that underground nematode infection significantly enhances the defense ability of tomato plants against whitefly.     

Phloem-feeding herbivory on flowering melon plants enhances attraction of parasitoids by shifting floral to defensive volatiles

Emission of herbivore-induced plant volatiles (HIPVs) can differ according to the type of herbivory and the plant development stage, ultimately affecting recruitment of the natural enemy. Little is known about plant defenses induced at the flowering stage by phloem-feeding insects. We investigated the olfactory preference of Encarsia desantisi parasitoids and the chemical profile of flowering melon plants induced or not by the phloem-feeding of Bemisia tabaci whiteflies. In addition, we tested whether the parasitoids were attracted to synthetic defensive HIPVs, which mimicked whitefly-infested flowering melons. The parasitoids recognized volatiles from undamaged melons but preferred the scent of host-infested melons in olfactometry assays. Amounts of most individual volatiles did not differ between plant treatments; however, only whitefly-induced melons released methyl salicylate and tetradecane, compounds known to attract parasitoids. Interestingly, grouping volatiles by chemical classes revealed that whitefly-infested melon released larger amounts of monoterpenes and smaller amounts of benzenoids than undamaged melons, which might underlying the parasitoid attraction and indicate a possible trade-off between defensive and reproductive defenses at the melon flowering stage. Additionally, E. desantisi preferred the mix of synthetic and defensive HIPVs over hexane (control), opening a new avenue for further investigations in using olfactory lures for B. tabaci biological control. This study is the first report of induced defenses in melon plants and their mediation in a tritrophic interaction, as well as the first record of E. desantisi behavioral preference for HIPVs.     

Acylsucrose-producing tomato plants forces Bemisia tabaci to shift its preferred settling and feeding site

Background

The whitefly Bemisia tabaci (Genn.) causes dramatic damage to plants by transmitting yield-limiting virus diseases. Previous studies proved that the tomato breeding line ABL 14-8 was resistant to B. tabaci, the vector of tomato yellow leaf curl disease (TYLCD). This resistance is based on the presence of type IV glandular trichomes and acylsucrose production. These trichomes deter settling and probing of B. tabaci in ABL 14-8, which reduces primary and secondary spread of TYLCD.

Methodology/Principal Findings

Whitefly settlement preference was evaluated on the adaxial and abaxial leaf surfaces of nearly-isogenic tomato lines with and without B. tabaci-resistance traits, ‘ABL 14-8 and Moneymaker’ respectively, under non-choice and free-choice conditions. In addition, the Electrical Penetration Graph technique was used to study probing and feeding activities of B. tabaci on the adaxial and abaxial leaf surfaces of the same genotypes. B. tabaci preferred to settle on the abaxial than on the adaxial surface of ‘Moneymaker’ leaves, whereas no such preference was observed on ABL 14-8 tomato plants at the ten-leaf growth stage. Furthermore, B. tabaci preferred to feed on the abaxial than on the adaxial leaf surface of ‘Moneymarker’ susceptible tomato plants as shown by a higher number of sustained phloem feeding ingestion events and a shorter time to reach the phloem. However, B. tabaci standard probing and feeding behavior patterns were altered in ABL 14-8 plants and whiteflies were unable to feed from the phloem and spent more time in non-probing activities when exposed to the abaxial leaf surface.

Conclusions/Significance

The distorted behavior of B. tabaci on ABL 14-8 protects tomato plants from the transmission of phloem-restricted viruses such as Tomato yellow leaf curl virus (TYLCV), and forces whiteflies to feed on the adaxial side of leaves where they feed less efficiently and become more vulnerable to natural enemies.