BTH-treated Plants Research Articles

Elicitor-Driven Defense Mechanisms: Shielding Cotton Plants against the Onslaught of Cotton Leaf Curl Multan Virus (CLCuMuV) Disease.

Salicylic acid (SA), benzothiadiazole (BTH), and methyl jasmonate (MeJA) are potential elicitors found in plants, playing a crucial role against various biotic and abiotic stresses. The systemic acquired resistance (SAR) mechanism was evaluated in cotton plants for the suppression of Cotton leaf curl Multan Virus (CLCuMuV) by the exogenous application of different elicitors. Seven different treatments of SA, MeJA, and BTH were applied exogenously at different concentrations and combinations. In response to elicitors treatment, enzymatic activities such as SOD, POD, CAT, PPO, PAL, β-1,3 glucanse, and chitinase as biochemical markers for resistance were determined from virus-inoculated and uninoculated cotton plants of susceptible and tolerant varieties, respectively. CLCuMuV was inoculated on cotton plants by whitefly (Bemesia tabaci biotype Asia II-1) and detected by PCR using specific primers for the coat protein region and the Cotton leaf curl betasatellite (CLCuMuBV)-associated component of CLCuMuV. The development of disease symptoms was observed and recorded on treated and control plants. The results revealed that BTH applied at a concentration of 1.1 mM appeared to be the most effective treatment for suppressing CLCuMuV disease in both varieties. The enzymatic activities in both varieties were not significantly different, and the disease was almost equally suppressed in BTH-treated cotton plants following virus inoculation. The beta satellite and coat protein regions of CLCuMuV were not detected by PCR in the cotton plants treated with BTH at either concentration. Among all elicitors, 1.1 mM BTH was proven to be the best option for inducing resistance after the onset of CLCuMuV infection and hence it could be part of the integrated disease management program against Cotton leaf curl virus.

Effects of a Salicylic Acid Analog on Aphis gossypii and Its Predator Chrysoperla carnea on Melon Plants

The salicylic acid analog BTH (benzo-(1,2,3)-thiadiazole-7-carbothioic-acid S-methyl ester) induces systemic acquired resistance by promoting plant resistance against numerous plant pathogens and some insect pests. The objective of the research was to evaluate the activation of plant defenses with BTH on melon (Cucumis melo L., Cucurbitaceae) and its effects on the herbivore Aphis gossypii Glover, 1877 (Hemiptera: Aphididae) and on the aphid predator Chrysoperla carnea (Stephens, 1836) (Neuroptera: Chrysopidae). Under laboratory conditions, plants were sprayed with BTH (50 g/ha) zero (B0), four (B4), and seven (B7) days prior exposure to insects. B0 treatment resulted in 100% mortality of aphid nymphs and disrupted adult feeding behavior (recorded by electrical-penetration-graphs technique), by prolonging the time to reach the phloem, requiring more probes to first salivation and reducing ingestion activities. There were no effects on feeding behavior of A. gossypii fed on B4 plants but on its life history because fewer nymphs were born, intrinsic rate of natural growth decreased, and mortality increased. There were no effects on biological parameters of aphids reared on B7 plants. Prey consumption by C. carnea larvae when predated A. gossypii fed on BTH-treated plants was not different among treatments. Therefore, BTH enhances the suppression of A. gossypii in the short term, without negative effects on the predatory larva C. carnea, which makes this plant strengthener a useful tool to be considered in integrated pest management programs.

Benzothiadiazole and nitrogen source modify the nitrogen metabolism in cucumber infected with Pseudomonas syringae pv. lachrymans

The influence of nitrogen source and benzothiadiazole (BTH) pretreatment on nitrogen metabolism-related enzyme activities was studied in cucumber plants infected with Pseudomonas syringae pv. lachrymans (Psl). The plants were grown hydroponically in media containing nitrate and urea at different ratios: 100% NO3− (100N), 75% NO3−/25% urea (75N) and 50% NO3−/50% urea (50N) and pretreated with BTH prior to Psl inoculation.In the presence of urea in the nutritional medium the activities of most enzymes in the control plants decreased. Disease development was influenced by nitrogen nutrition and the weakest symptoms occurred in the 75N plants. The BTH-induced disease restriction was dependent on the type of nitrogen nutrition and was most visible in the 50N plants (about 2-fold reduction of infected leaf area compared to the plants not pretreated with BTH). The nitrogen metabolism was modified by BTH and this effect was stronger than that of urea. Irrespective of the type of nitrogen nutrition, in the BTH-treated plants, the activities of nitrate reductase, ferredoxin- and NADH-dependent glutamate synthase decreased whereas those of NADH-dependent glutamate dehydrogenase (NADH-GDH), aminotransferases and NADP-dependent isocitrate dehydrogenase were enhanced compared to the controls. After infection, in all nutritional variants, the strongest effect of BTH pretreatment was observed for NADH-GDH on the 2 day, when its activity increased by 85–256% compared to the infected non-pretreated plants. Our results concerning the glutamate-metabolizing enzymes suggest that BTH treatment combined with low dose of urea may have a positive influence on the nitrogen management leading to the inhibition of disease symptom progress.

Priming of cowpea volatile emissions with defense inducers enhances the plant's attractiveness to parasitoids when attacked by caterpillars.

The manipulation of herbivore-induced volatile organic compounds (HI-VOCs) via the application of the inducers benzo(1,2,3)thiadiazole-7-carbothioic acid S-methyl ester (BTH) and laminarin (β-1,3-glucan) is known to enhance the attractiveness of caterpillar-damaged cotton and maize plants to parasitoids. To test if this is also the case for legumes, we treated cowpea (Vigna unguiculata var. unguiculata) with these inducers and studied the effects on HI-VOC emissions and the attraction of three generalist endoparasitoids. After the inducers had been applied and the plants subjected to either real or mimicked herbivory by Spodoptera littoralis caterpillars, females of the parasitoids Campoletis sonorensis and Microplitis rufiventris showed a strong preference for BTH-treated plants, whereas Cotesia females were strongly attracted to both BTH- and laminarin-treated plants with real or mimicked herbivory. Treated plants emitted more of certain HI-VOCs, but considerably less indole and linalool and less of several sesquiterpenes. Multivariate data analysis revealed that enhanced wasp attraction after treatment was correlated with high relative concentrations of nonanal, α-pinene, (E)-β-ocimene and (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), and with low relative concentrations of indole, (S)-linalool and (E)-β-farnesene. Inducer treatments had no significant effect on leaf consumption by the caterpillars. Our findings confirm that treating cowpea plants with inducers can enhance their attractiveness to biological control agents. © 2017 Society of Chemical Industry.

Root-mediated signal transmission of systemic acquired resistance against above-ground and below-ground pathogens.

Background and Aims Plants modulate defence signalling networks in response to various biotic stresses via inter-organ communications. The root-mediated transmission of systemic acquired resistance (SAR) against soil-borne and air-borne plant pathogens from SAR-induced plants to neighbouring plants subjected to local chemical and pathogen treatments was evaluated. Methods The first two plants out of ten Nicotiana benthamiana seedlings were pre-treated with the SAR-triggering chemical benzothiadiazole (BTH). All ten seedlings were then challenged with two pathogenic bacteria, i.e. the root (bacterial wilt) pathogen Ralstonia solanacearum and the leaf (wildfire) pathogen Pseudomonas syringae pv. tabaci, at 7 d after SAR induction. Key Results Disease severity was noticeably lower in BTH-pre-treated plants than in the control. Surprisingly, two plants located next to BTH-treated plants exhibited reduced disease symptoms indicating that SAR signal transmission occurred through the root system. Determinant(s) secreted from the root system were search for and it was found that salicylic acid (SA) is a major molecule involved in SAR transmission through the root. Analysis of the expression of the defence-related genes N. benthamiana pathogenesis-related gene 1a (NbPR1a) and NbPR2 confirmed that BTH treatment elicited SAR via root-root transmission between plants. Plants with knock-down of the multiple resistance component SGT1 and SA biosynthesis-related gene ICS1 by Tobacco rattle virus-mediated virus-induced gene silencing exhibited a lack of root-mediated SAR transmission. The biological relevance of this finding was validated by challenge with the SAR-inducing avirulent pathogen P. syringae pv. syringae instead of BTH, which produced similar results. Conclusions Our findings demonstrated that SAR is transmissible through the root system from SAR-triggered plants to neighbouring plants.

Nitrogen forms modulate effects of benzothiadiazole and arbutin on cucumber sugar metabolism

The effect of benzothiadiazole (BTH) and arbutin (ARB) on sugar metabolism and plant fitness in cucumber growing hydroponically in media with different doses of NO3 - and urea as nitrogen sources (100 % NO3 -, 75 % NO3 - + 25 % urea, and 50 % NO3 - + 50 % urea) was studied on the 7th and 14th day after the treatment. The glucose, sucrose, and chlorophyll (Chl) content, acid and alkaline invertases and lactate dehydrogenase activities, as well as leaf area of the 3rd and 5th leaves were determined. Urea changed the plant sugar metabolism in a dose-, time- and leaf-age-dependent manners and influenced a cucumber response to the BTH and ARB treatments. The BTH caused a significant cessation of growth, a decrease in Chl content, a reduction of leaf area, and an enhancement of lactate dehydrogenase and alkaline invertase activities. In the older leaves of the BTH-treated plants, a high accumulation of glucose and sucrose was found. At the lower dose of urea, the metabolic changes were limited. In the ARB-treated plants, the Chl content remained unchanged in all the nitrogen variants. In these plants, decrease in glucose and sucrose content and in both invertase activities was observed mainly in younger leaves of the plants grown on the high dose of urea. The ARB improved the fitness of the cucumber plants grown in the presence of urea.

WRKY45-dependent priming of diterpenoid phytoalexin biosynthesis in rice and the role of cytokinin in triggering the reaction.

Plant activators such as benzothiadiazole (BTH) protect plants against diseases by priming the salicylic acid (SA) signaling pathway. In rice, the transcription factor WRKY45 plays a central role in this process. To investigate the mechanism involved in defense-priming by BTH and the role of WRKY45 in this process, we analyzed the transcripts of biosynthetic genes for diterpenoid phytoalexins (DPs) during the rice–Magnaporthe oryzae interaction. The DP biosynthetic genes were barely upregulated in BTH-treated rice plants, but were induced rapidly after M. oryzae infection in a WRKY45-dependent manner. These results indicate that the DP biosynthetic genes were primed by BTH through WRKY45. Rapid induction of the DP biosynthetic genes was also observed after M. oryzae infection to WRKY45-overexpressing (WRKY45-ox) plants. The changes in gene transcription resulted in accumulation of DPs in WRKY45-ox and BTH-pretreated rice after M. oryzae infection. Previously, we reported that cytokinins (CKs), especially isopentenyladenines, accumulated in M. oryzae-infected rice. Here, we show that DP biosynthetic genes are regulated by the SA/CK synergism in a WRKY45-dependent manner. Together, we propose that CK plays a role in mediating the signal of M. oryzae infection to trigger the induction of DP biosynthetic genes in BTH-primed plants.Electronic supplementary materialThe online version of this article (doi:10.1007/s11103-014-0221-x) contains supplementary material, which is available to authorized users.

Induction of Salicylic Acid–Mediated Defense Response in Perennial Ryegrass Against Infection by Magnaporthe oryzae

Incorporation of plant defense activators is an innovative approach to development of an integrated strategy for the management of turfgrass diseases. The effects of salicylic acid (SA), benzothiadiazole (BTH, chemical analog of SA), jasmonic acid (JA), and ethephon (ET, an ethylene-releasing compound) on development of gray leaf spot in perennial ryegrass (Lolium perenne L.) caused by Magnaporthe oryzae were evaluated. Gray leaf spot disease incidence and severity were significantly decreased when plants were treated prior to inoculation with SA, BTH, and partially by ET but not by JA. Accumulation of endogenous SA and elevated expression of pathogenesis-related (PR)-1, PR-3.1, and PR-5 genes were associated with inoculation of plants by M. oryzae. Treatment of plants with SA enhanced expression levels of PR-3.1 and PR-5 but did not affect the PR-1 level, whereas BTH treatment enhanced relative expression levels of all three PR genes. Microscopic observations of leaves inoculated with M. oryzae revealed higher frequencies of callose deposition at the penetration sites in SA- and BTH-treated plants compared with the control plants (treated with water). These results suggest that early and higher induction of these genes by systemic resistance inducers may provide perennial ryegrass with a substantial advantage to defend against infection by M. oryzae.

Benzothiadiazole-elicited defense priming and systemic acquired resistance against bacterial and viral pathogens of pepper under field conditions

Like the innate immunity in mammals, plants have developed an induced resistance, referred to as systemic acquired resistance (SAR). Recently, defense priming that is not related to the direct activation of defenses, but instead elicits more rapid induction of resistance mechanisms following trigger application, has been proposed to explain the long-lasting effect of SAR. However, the majority of previous studies have focused on understanding the molecular mechanism underlying priming under in vitro and laboratory conditions. This study examined whether defense priming occurred and was detectable with SAR marker genes by a chemical elicitor, benzothiadiazole (BTH), under field conditions. Pepper seedling application of 0.5 mM BTH was sufficient to prime the CaPR4 gene for 20 days as well as to induce SAR against bacterial spot caused by Xanthomonas axonopodis. Transcriptome analysis revealed to prime defense hormonal signaling and antimicrobial compound production genes. At the end of the season, when bacterial spot and Cucumber mosaic virus disease outbreaks naturally occurred, BTH-treated plants demonstrated less disease symptoms. Our results indicate that the priming of SAR genes plays a critical role in plant protection against pathogens under natural conditions.

Trichoderma harzianum T39 induces resistance against downy mildew by priming for defense without costs for grapevine

Downy mildew caused by Plasmopara viticola is one of the most destructive grapevine diseases worldwide. The biocontrol agent Trichoderma harzianum T39 (T39) has previously been shown to be an inducer of grapevine resistance, and we aimed at characterizing the molecular mechanisms activated by T39 and the energy costs of the induced resistance in terms of plant growth. Here, we showed that T39 reduced downy mildew severity on susceptible grapevines under controlled greenhouse conditions by a direct modulation of defense-related genes and the activation of priming for enhanced expression of these genes after pathogen inoculation. The stronger local than systemic modulation of defense-related genes corresponded to an higher local than systemic disease control in T39-treated plants. The activation of a priming state was confirmed by the absence of any negative effect of T39 on grapevine growth, shoot and root weight, leaf dimension and chlorophyll content, in contrast to benzothiadiazole (BTH) applications. Priming of defense gene expression was greater in T. harzianum T39- than in BTH-treated plants. The modulation of marker genes suggested the involvement of jasmonic acid and ethylene signals in the defense processes induced by T39, in contrast to the salicylic acid pathway activated by BTH. These results offer a greater understanding of the mechanisms underlying the grapevine induced resistance and indicate that T. harzianum T39 can be used to control downy mildew without apparent costs for grapevine growth.

U romyces appendiculatus Infection in BTH-Treated Bean Plants: Ultrastructural Details of a Lost Fight

The mechanisms of BTH [benzo-(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester]-induced resistance against bean rust caused by Uromyces appendiculatus have been explored in Phaseolus vulgaris by light and transmission electron microscopy, following the infection progression in plants challenged 7 days after treatment. While BTH did not affect uredospore germination and fungal penetration in the substomatal cavity, a first impairment to the colonization appeared evident about 48-96 h after inoculation, with alterations of infection hypha structure and reduction in mycelium expansion. No differences were found in this phase regarding the formation and ultrastructure of haustoria in untreated and BTH-treated plants, except for the deposition of electron-opaque material in the extrahaustorial matrix of the latter. A second and decisive impairment in fungal progression was observed at 7-10 days after inoculation when host cell penetrated, or in close contact with the fungal hyphae, were impregnated by phenolic compounds. The same was observed in fungal walls, particularly around haustoria, thus hampering the biotrophic habitus of the fungus and further mycelium spreading. This, in turn, prevented the evasion of fungal reproductive structures, the uredinia, and the appearance of visible symptoms. No particular ultrastructural alterations were observed in most of the penetrated cells, even at late stages of infection, indicating that BTH treatment does not induce host cells to respond with a hypersensitive reaction (HR). A parallel time course of the expression of phenylalanine ammonia lyase (PAL) gene, the key enzyme for the synthesis of phenylpropanoidic phytoalexins and many other phenolics, has shown that PAL mRNA is strongly and persistently transcripted in BTH-treated plants since the 6th h after treatment, though no apparent ultrastructural alterations were detectable up to some days after pathogen challenging. This indicates that BTH, at the employed concentration of 0.3 mM, directly activates the plant's own defences, thus accounting for the observed full protection against bean rust.

Activity of benzothiadiazole on chrysanthemum yellows phytoplasma (‘ Candidatus Phytoplasma asteris’) infection in daisy plants

A commercial preparation of the plant resistance elicitor benzothiadiazole (BTH) (Bion, Syngenta Crop Protection) was tested for its capacity to induce systemic resistance against chrysanthemum yellow phytoplasma (CYP) infection in the Chrysanthemum carinatum plant. Following one 2.4 mM BTH application, plants were exposed to CYP-infective Macrosteles quadripunctulatus leafhoppers. Symptom development and phytoplasma multiplication in the test plants were measured over time. BTH application delayed symptom development and phytoplasma multiplication in treated plants compared with the control ones. CYP titre and symptom severity were significantly lower for the first two weeks post-inoculation in treated plants compared with the control ones, suggesting that systemic acquired resistance (SAR) induced by BTH in C. carinatum is temporary. Higher concentrations of BTH resulted in phytotoxic effects involving the whitening of apical leaves. BTH application was ineffective in reducing the transmission efficiency of CYP by its leafhopper vector. Actually, in both single and group transmission tests, the proportion of infected plants was similar among BTH-treated and untreated plants. The survival of M. quadripunctulatus was unaffected by feeding on BTH-treated daisy plants. Moreover, when leafhoppers were allowed to choose between treated and untreated plants, they showed no preference. We conclude that SAR induced in daisy plants by BTH has no detrimental effects on the vector leafhopper. If the activity of BTH against phytoplasmas is confirmed also on other phytoplasma/host–plant associations, BTH applications might be included in new, more environmentally friendly, integrated management strategies of phytoplasmoses.

BTH-mediated antioxidant system responses in apple leaf tissues

BTH (S-methylbenzo-1,2,3-thiadiazole-7-carbothiate), an active compound of the commercial preparation Bion, has been studied as an elicitor of resistance to fire blight ( Erwinia amylovora) in apple. However, the biochemical mechanisms of its action are not fully elucidated. Our study indicated that BTH at the best time of its protection activity (2–14 days after application) induced changes in prooxidant–antioxidant balance in the leaves of apple trees, but in different ways in the enzymatic and nonenzymatic antioxidants. Glutathione as low molecular antioxidant as well as superoxide anion radical and lipid peroxides as oxidants exhibited changes at the early phase of BTH action. Glutathione-dependent enzymes were strongly affected by the elicitor used. On the 2nd day glutathione transferase (GST) and glutathione peroxidase (GSH-Px) activities increased by about 70% and 30% above the control, respectively. GST activity normalized about the 14th day but GSH-Px at the same time showed 27% of the control value. Among enzymes utilising hydrogen peroxide only catalase showed increase (37%) at the early phase of experiment. Compared with the control, BTH-treated plants did not show changes in ascorbate peroxidase and phenylalanine ammonia-lyase activities. Tocopherol (TOC) level diminished starting from the 7th day after BTH treatment and on the 14th day it was only 28% of the control. It is proposed that extinguishing of BTH-mediated signal resulted from TOC and glutathione action. The diminished ascorbate level at all examined times may play a crucial role in BTH-mediated cell growth regulation. The direct influence of BTH on lipid metabolism should be also taken into consideration.

Benzo-(1,2,3)-thiadiazole-7-carbothioic S-methyl ester (BTH) stimulates defense reactions in Xanthosoma sagittifolium

The root rot disease caused by Pythium myriotylum is responsible for about 70% of cocoyam production loss in Cameroon. The potential of benzo-(1,2,3)-thiadiazole-7-carbothioic S-methyl ester (BTH) to trigger resistance in cocoyam (Xanthosoma sagittifolium) plants against P. myriotylum was investigated. Under controlled conditions, BTH was an efficient elicitor of some defense reactions in cocoyam. Application of 0.2 mg ml−1 of BTH on leaves 7 days before inoculation of roots with P. myriotylum enhanced the activities of peroxidase (Pox) and polyphenoloxidase (PPO) as well as the total phenolic content. This resistance was noted as a decrease in disease incidence and severity in BTH-treated plants. This increase in Pox activities was correlated with two new isoforms in a white (sensitive) cultivar inoculated after stimulation. In a yellow (resistant) cultivar, stimulation was characterized by the appearance of one isoform. Qualitative analysis of phenolic compounds by HPLC showed an increase of hydroxycinnamic and flavonoid derivatives after inoculation. We also observed the appearance of a new caffeoylshikimic acid derivative after stimulation followed by inoculation of both cultivars. The findings indicated that the pattern of induction is different and depends on the variety.

Airborne induction and priming of plant defenses against a bacterial pathogen.

Herbivore-induced plant volatiles affect the systemic response of plants to local damage and hence represent potential plant hormones. These signals can also lead to "plant-plant communication," a defense induction in yet undamaged plants growing close to damaged neighbors. We observed this phenomenon in the context of disease resistance. Lima bean (Phaseolus lunatus) plants in a natural population became more resistant against a bacterial pathogen, Pseudomonas syringae pv syringae, when located close to conspecific neighbors in which systemic acquired resistance to pathogens had been chemically induced with benzothiadiazole (BTH). Airborne disease resistance induction could also be triggered biologically by infection with avirulent P. syringae. Challenge inoculation after exposure to induced and noninduced plants revealed that the air coming from induced plants mainly primed resistance, since expression of PATHOGENESIS-RELATED PROTEIN2 (PR-2) was significantly stronger in exposed than in nonexposed individuals when the plants were subsequently challenged by P. syringae. Among others, the plant-derived volatile nonanal was present in the headspace of BTH-treated plants and significantly enhanced PR-2 expression in the exposed plants, resulting in reduced symptom appearance. Negative effects on growth of BTH-treated plants, which usually occur as a consequence of the high costs of direct resistance induction, were not observed in volatile organic compound-exposed plants. Volatile-mediated priming appears to be a highly attractive means for the tailoring of systemic acquired resistance against plant pathogens.

Benzothiadiazole affects the leaf proteome in arctic bramble (Rubus arcticus).

Benzothiadiazole (BTH) induces resistance to the downy mildew pathogen, Peronospora sparsa, in arctic bramble, but the basis for the BTH-induced resistance is unknown. Arctic bramble cv. Mespi was treated with BTH to study the changes in leaf proteome and to identify proteins with a putative role in disease resistance. First, BTH induced strong expression of one PR-1 protein isoform, which was also induced by salicylic acid (SA). The PR-1 was responsive to BTH and exogenous SA despite a high endogenous SA content (20-25 microg/g fresh weight), which increased to an even higher level after treatment with BTH. Secondly, a total of 792 protein spots were detected in two-dimensional gel electrophoresis, eight proteins being detected solely in the BTH-treated plants. BTH caused up- or down-regulation of 72 and 31 proteins, respectively, of which 18 were tentatively identified by mass spectrometry. The up-regulation of flavanone-3-hydroxylase, alanine aminotransferase, 1-aminocyclopropane-1-carboxylate oxidase, PR-1 and PR-10 proteins may partly explain the BTH-induced resistance against P. sparsa. Other proteins with changes in intensity appear to be involved in, for example, energy metabolism and protein processing. The decline in ATP synthase, triosephosphate isomerase, fructose bisphosphate aldolase and glutamine synthetase suggests that BTH causes significant changes in primary metabolism, which provides one possible explanation for the decreased vegetative growth of foliage and rhizome observed in BTH-treated plants.

Benzothiadiazole Induces the Accumulation of Phenolics and Improves Resistance to Powdery Mildew in Strawberries

Benzothiadiazole (BTH) enhanced the accumulation of soluble and cell-wall-bound phenolics in strawberry leaves and also improved the resistance to powdery mildew infection under greenhouse conditions. The most pronounced change was seen in the levels of ellagitannins, which increased up to 2- to 6-fold 4 days after the BTH application, but persisted only in the inoculated plants. The induction of phenolic metabolism by BTH was also reflected in the fruits, several compounds being increased in inoculated, BTH-treated plants. Basal salicylic acid (SA) content was high in strawberry leaves, but increased in a similar fashion to other phenolics after the treatments. Several phenolic compounds were identified in strawberries for the first time. For example, ellagic acid deoxyhexose, three agrimoniin-like ellagitannins, sanguiin H-10- and lambertianin C-like ellagitannins in the leaves, ellagic acid, p-coumaric acid, gallic acid, and kaempferol hexose in the cell-wall-bound fraction of the leaves, and kaempferol malonylglucoside in the fruits. The findings show that BTH can enhance the accumulation of phenolics in strawberry plants which may then be involved in the BTH-induced resistance to powdery mildew.

From Field to Health: A Simple Way To Increase the Nutraceutical Content of Grape As Shown by NO-Dependent Vascular Relaxation

Polyphenolic grapevine components involved in plant resistance against pathogens possess various pharmacological properties that include nitric oxide (NO)-dependent vasodilation and anti-inflammatory and free radical scavenging activities, which may explain the protective effect of moderate red wine consumption against cardiovascular disease. The aim of this work was (a) to verify the possibility that preharvest treatments of grapevine with a plant activator, benzo-(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester (BTH), could lead to an enriched nutraceutical potential of wine and (b) to characterize the profile of metabolites responsible for pharmacological activity. Plant spraying at the end of veraison, with a water suspension of BTH (0.3 mM), led to increased whole anthocyanin content as confirmed by HPLC comparative analysis. Extracts from berry skins of BTH-treated grapevines caused NO-dependent vasorelaxation, with a concentration-response curve that was significantly shifted to the left of the control non-BTH-treated curve. Moreover, 1:1000 dilutions of berry extracts from BTH-treated plants significantly increased basal production of guanosine 3',5'-cyclic monophosphate (cGMP) in human vascular endothelial cells when compared to the corresponding extracts of untreated plants. These results show that BTH treatment increases anthocyanin content of grape extracts, as well as their ability to induce NO-mediated vasoprotection. No increase of anthocyanin content was observed in the wine extracts from BTH-treated vines. It is concluded that BTH treatment could be exploited to increase the nutraceutical potential of grapes.

Benzothiadiazole Induces Local Resistance to Bemisia tabaci (Hemiptera: Aleyrodidae) in Tomato Plants

Plant resistance to the B and Q biotypes of sweetpotato whitefly, Bemisa tabaci (Gennadius), induced by benzo [1,2,3] thiadiazole-7-carbothioic acid-S-methyl ester (BTH or acibenzolar-S-methyl) in tomato 'Marmande' plants was evaluated in free-choice and no-choice assays under different conditions. BTH is the active ingredient of the Syngenta plant activator Bion. BTH treatment affected host preference of B. tabaci (B and Q biotypes) adults on plants sprayed with Bion at 0.2 and 0.4 g/liter during the earlier days of free-choice assays. As a consequence, a decrease in the total number of eggs (although female fecundity was not affected) and in the final number of pupae and empty pupal cases was observed. The effect produced by BTH applied at 0.1 g/liter Bion was not significant. In no-choice assays, a reduction of the numbers of first-stage larvae and total individuals and a delay in insect development were observed when local treatment was restricted to one leaflet per plant, 5 d before B. tabaci (biotype B) infestation. This acquired resistance induced by BTH seemed to be locally expressed because of the differences between treated and nontreated leaflets in the same plants, whereas no differences in nontreated leaflets were observed between BTH-treated and control plants.

Benzothiadiazole enhances resveratrol and anthocyanin biosynthesis in grapevine, meanwhile improving resistance to Botrytis cinerea.

Pre-harvest multiple treatments of grapevine (cv. Merlot) with the plant activator benzothiadiazole (BTH, 0.3 mM) enhanced trans-resveratrol content in berries by about 40%. An even more striking effect was observed on anthocyanin synthesis, particularly on malvidine 3-glucoside, malvidine 3-(6-O-acetyl)glucoside and malvidine 3-(6-O-p-coumaroyl)glucoside, whose amounts were more than doubled. These data were obtained with a novel and time-saving HPLC method, set up for the simultaneous detection of stilbenes and anthocyanins, using an RF-10Axl fluorimetric detector instrument, with excitation at 330 nm and emission at 374 nm, and a SPD-Avp UV detector with absorption at 520 nm. Furthermore, BTH treatments induced systemic acquired resistance in grapevine, as assessed by inoculating clusters from treated and untreated plants with Botrytis cinerea. Disease severity, estimated according to the percentage of infected berries per cluster, was significantly reduced in grapes from BTH-treated plants. These results indicate that BTH treatments, besides improving the content of two important classes of nutraceuticals, with their well-known antioxidant, antitumoral, and phytoestrogenic activities, could be exploited in vineyard to protect grape against gray mould infection, thereby limiting an excessive use of fungicides