Validation of RNA interference in western corn rootworm Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae) adults

BACKGROUND: RNA interference (RNAi) is commonly used in insect functional genomics studies and usually involves direct injection of double‐stranded RNA (dsRNA). Only a few studies have involved exposure to dsRNAs through feeding. For western corn rootworm (Diabrotica virgifera virgifera) larvae, ingestion of dsRNA designed from the housekeeping gene, vacuolar ATPase (vATPase) triggers RNAi causing growth inhibition and mortality; however, the effect of dsRNA feeding on adults has not been examined. In this research, WCR adults were fed with vATPase‐dsRNA‐treated artificial diet containing a cucurbitacin bait, which is a proven feeding stimulant for chrysomelid beetles of the subtribe Diabroticina to which rootworms belong. RESULTS: Real‐time PCR confirmed suppression of vATPase expression and western blot analysis indicated reduced signal of a protein that cross‐reacted with a vATPase polyclonal antiserum in WCR adults exposed to artificial diet treated with dsRNA and cucurbitacin bait. Continuous feeding on cucurbitacin and dsRNA‐treated artificial diet resulted in more than 95% adult mortality within 2 weeks while mortality in control treatments never exceeded 20%. CONCLUSIONS: This research clearly demonstrates the effect of RNAi on WCR adults that have been exposed to dsRNA by feeding and establishes a tool to screen dsRNAs of potential target genes in adults. This technique may serve as an alternative to target screening of larvae which are difficult to maintain on artificial diets. Copyright © 2011 Society of Chemical Industry     

Resistance to pyraclostrobin, boscalid and multiple resistance to Pristine® (pyraclostrobin + boscalid) fungicide in Alternaria alternata causing alternaria late blight of pistachios in California

Pristine® (pyraclostrobin + boscalid) is a fungicide registered for the control of alternaria late blight in pistachio. A total of 95 isolates of Alternaria alternata collected from orchards with and without a prior history of Pristine® sprays were tested for their sensitivity towards pyraclostrobin, boscalid and Pristine® in conidial germination assays. The EC₅₀ values for 35 isolates from orchards without Pristine® sprays ranged from 0·09 to 3·14 µg mL^?1 and < 0·01 to 2·04 µg mL^?1 for boscalid and Pristine®, respectively. For pyraclostrobin, 27 isolates had EC₅₀ < 0·01 µg mL^?1 and six had low resistance (mean EC₅₀ value = 4·71 µg mL^?1). Only one isolate was resistant to all three fungicides tested, with EC₅₀ > 100 µg mL^?1. Among 59 isolates from the orchard with a history of Pristine® sprays, 56 were resistant to pyraclostrobin; only two were sensitive (EC₅₀ < 0·01 µg mL^?1) and one was weakly resistant (EC₅₀ = 10 µg mL^?1). For the majority of these isolates EC₅₀ values ranged from 0·06 to 4·22 µg mL^?1 for boscalid and from 0·22 to 7·74 µg mL^?1 for Pristine®. However, seven isolates resistant to pyraclostrobin were also highly resistant to boscalid and Pristine® and remained pathogenic on pistachio treated with Pristine®. Whereas strobilurin resistance is a common occurrence in Alternaria of pistachio, this is the first report of resistance to boscalid in field isolates of phytopathogenic fungi. No cross resistance between pyraclostrobin and boscalid was detected, suggesting that Pristine® resistance appears as a case of multiple resistance.     

Electrical penetration graph evidence that pymetrozine toxicity to the rice brown planthopper is by inhibition of phloem feeding

BACKGROUND: Pymetrozine is a valuable novel insecticide for control of sucking insects, including the brown planthopper Nilaparvata lugens (Stål), one of the most serious pests on rice. This study was conducted to elucidate the action mechanisms of pymetrozine on the feeding behaviour of the planthopper. RESULTS: The activity test showed that pymetrozine primarily functioned as an antifeedant that caused starvation and death in N. lugens, rather than having neurotoxicity. Pymetrozine‐treated insects died at a significantly slower speed than insects treated with starvation. Electrical penetration graph (EPG) data indicated that pymetrozine significantly increased the duration of non‐probing periods and had a strong inhibition to phloem ingestion. The inhibition was strongly dose dependent, resulting in a complete suppression of the activity in the phloem region when the pymetrozine concentration was increased to 400 mg L^?1. Starvation caused by inhibition of phloem ingestion might be a major toxicity mechanism of pymetrozine. EPG data also showed that pymetrozine had no significant effect on stylet movement and duration of xylem sap ingestion. CONCLUSION: The study revealed that pymetrozine disturbed the feeding behaviour of N. lugens mainly by increasing the non‐probe period and inhibiting phloem ingestion. The inhibition resulted in a slow death similar to starvation. Copyright © 2011 Society of Chemical Industry     

Composition and biological activity of essential oils from Labiatae against Nezara viridula (Hemiptera: Pentatomidae) soybean pest

BACKGROUND: Plant essential oils have been recognised as an important natural source of insecticide. This study analysed the chemical constituents and bioactivity of essential oils that were isolated via hydrodistillation from Origanum vulgare L. (oregano) and Thymus vulgaris L. (thyme) against eggs, second instar and adults of Nezara viridula (L.). RESULTS: The major component of oregano was p‐cymene, and, for thyme, thymol. The ovicidal activity was tested by topical application; the essential oil from thyme was more effective. The fumigant activity was evaluated in an enclosed chamber; the LC₅₀ values for oregano were 26.8 and 285.6 µg mL^?1 for nymphs and adults respectively; for thyme they were 8.9 µg mL^?1 for nymphs and 219.2 µg mL^?1 for adults. To evaluate contact activity, a glass vial bioassay was used; the LC₅₀ values for oregano were 1.7 and 169.2 µg cm^?2 for nymphs and adults respectively; for thyme they were 3.5 and 48.8 µg cm^?2 respectively. The LT₅₀ analyses for contact and fumigant bioassays indicated that thyme was more toxic for nymphs and adults than oregano. Both oils produced repellency on nymphs and adults. CONCLUSION: These results showed that the essential oils from O. vulgare and T. vulgaris could be applicable to the management of N. viridula. Copyright © 2011 Society of Chemical Industry     

Synthesis of 2-amino-6-oxocyclohexenylsulfonamides and their activity against Botrytis cinerea

BACKGROUND: With the objective of exploring the fungicidal activity of 2‐oxocyclohexylsulfonamides (2), a series of novel 2‐amino‐6‐oxocyclohexenylsulfonamides (6 to 23) were synthesised, and their fungicidal activities against Botrytis cinerea Pers. were evaluated in vitro and in vivo. RESULTS: The compounds were characterised by IR, ¹H NMR and elemental analysis. Bioassay results of mycelial growth showed that compounds 6 to 23 had a moderate antifungal activity against B. cinerea. N‐(2‐methylphenyl)‐2‐(2‐methylphenylamino)‐4,4‐dimethyl‐6‐oxocyclohexenylsulfonamide (13) and N‐(2‐chlorophenyl)‐2‐(2‐chlorophenylamino)‐6‐oxocyclohexenylsulfonamide (21) showed best antifungal activities, with EC₅₀ values of 8.05 and 10.56 µg mL^?1 respectively. Commercial fungicide procymidone provided an EC₅₀ value of 0.63 µg mL^?1. The conidial germination assay showed that most of compounds 6 to 23 possessed excellent inhibition of spore germination and germ‐tube elongation of conidia of B. cinerea. For in vivo control of B. cinerea colonising cucumber leaves, the compound N‐cyclohexyl‐2‐(cyclohexylamino)‐4,4‐dimethyl‐6‐oxocyclohexenylsulfonamide (19) showed a better control effect than the commercial fungicide procymidone. CONCLUSION: The present work demonstrated that 2‐amino‐6‐oxocyclohexenylsulfonamides can be used as possible new lead compounds for further developing novel fungicides against B. cinerea. Copyright © 2011 Society of Chemical Industry     

Evaluation of alternative rice planthopper control by the combined action of oil-formulated Metarhizium anisopliae and low-rate buprofezin

BACKGROUND: High resistance of brown planthopper (BPH) Nilaparvata lugens Stål to common insecticides is a challenge for control of the pest. An alternative control strategy based on the combined application of fungal and chemical agents has been evaluated. RESULTS: Three gradient spore concentrations of oil‐formulated Metarhizium anisopliae (Metschnikoff) Sorokin (Ma456) were sprayed onto third‐instar nymphs in five bioassays comprising the low buprofezin rates of 0, 10, 20, 30 and 40 µg mL^?1 respectively. Fungal LC₅₀ after 1 week at 25 °C and 14:10 h light:dark photoperiod decreased from 386 conidia mm^?2 in the buprofezin‐free bioassay to 40 at the highest chemical rate. Buprofezin (LC₅₀: 1647, 486 and 233 µg mL^?1 on days 2 to 4) had no significant effect on the fungal outgrowths of mycosis‐killed cadavers at the low application rates. The fungal infection was found to cause 81% reduction in reproductive potential of BPH adults. In two 40 day field trials, significant planthopper (mainly BPH) control (54–60%) was achieved by biweekly sprays of two fungal candidates (Ma456 and Ma576) at 1.5 × 10¹³ conidia ha^?1 and elevated to 80–83% by incorporating 30.8 g buprofezin ha^?1 into the fungal sprays. CONCLUSION: The combined application of the fungal and chemical agents is a promising alternative strategy for BPH control. Copyright © 2010 Society of Chemical Industry     

Evaluation on the fitness and population projection of Nilaparvata lugens in response to elevated CO2 using two-sex life table

Abstract

The life history and fitness of Nilaparvata lugens being reared under ambient level (current CO₂ concentration), medially elevated (550?µL/L) and highly elevated (750?µL/L) CO₂ concentration for long-term generation were compared using two-sex life table instead of traditional age-specific life table. The results showed that significantly longer larval duration and lower fecundity of N. lugens were observed in 750?µL/L relative to 550?µL/L treatment (P?<?0.05). Accordingly, 550?µL/L CO₂ significantly enhanced population parameters of N. lugens, including significantly higher intrinsic rate of increase (r), finite rate (λ) and net reproductive rate (R_O), but not for 750?µL/L CO₂. Taken together, N. lugens performs enhanced development rate, fecundity and survival in individual life history and higher potential in population multiplication under 550?µL/L CO₂ level, while only indicates the less enhanced development rate and survival without significant increased capacity of population expansion under 750?µL/L CO₂ level. These results should facilitate predicting the fitness and potential population damage of N. lugens, which is valuable for the integrated control of N. lugens in the future increasing CO₂ concentration.     

Biocontrol traits of two Paenibacillus polymyxa strains SQR‐21 and WR‐2 in response to fusaric acid,a phytotoxin produced by Fusarium species

The aim of the present study was to investigate the effects of the phytotoxin fusaric acid (FA) on the biocontrol traits of two biocontrol strains Paenibacillus polymyxa WR‐2 and SQR‐21. The results showed that the growth of both WR‐2 and SQR‐21 decreased with increasing FA concentration, and at 70 and 80 μg mL^?1 FA, respectively, the strains were unable to grow. The biocontrol traits of both strains were negatively affected by FA concentration higher than 2·5 μg mL^?1. However, at 2·5 μg mL^?1 FA, biofilm formation and root colonization were not affected and there was even a positive effect on the production of spores and hydrolytic enzymes (protease and β‐l,3‐glucanase). The production of fusaricidin‐type antifungal compounds was increased with an increase in FA concentration up to 50 and 60 μg mL^?1 for WR‐2 and SQR‐21, respectively. The production of antifungal volatile organic compounds by WR‐2 and SQR‐21 was increased only at 2·5 μg mL^?1 FA. The effect of FA on the overall metabolic activity of WR‐2 and SQR‐21 was also determined. This study will help to understand the response of P. polymyxa strains to FA and will help to improve their biocontrol efficiency.     

Mefenoxam sensitivity and fitness analysis of Phytophthora nicotianae isolates from nurseries in Virginia, USA

Mefenoxam is one of the most commonly used fungicides for managing diseases caused by Phytophthora spp. on ornamentals. The objectives of this study were to determine whether Phytophthora nicotianae, a destructive pathogen of numerous herbaceous annual and perennial plant species in nurseries, has developed resistance to mefenoxam, and to evaluate the fitness of mefenoxam‐resistant isolates. Ninety‐five isolates of P. nicotianae were screened for sensitivity to mefenoxam on 20% clarified V8 agar at 100 a.i. µg mL^?1. Twenty‐five isolates were highly resistant to this compound with EC₅₀ values ranging from 235·2 to 466·3 µg mL^?1 and four were intermediately resistant with EC₅₀ values ranging from 1·6 to 2·9 µg mL^?1. Sixty‐six isolates were sensitive with EC₅₀ values less than 0·04 µg mL^?1. Nine resistant and seven sensitive isolates were tested for mefenoxam sensitivity on Pelargonium × hortorum cv. White Orbit. Mefenoxam provided good protection of pelargonium seedlings from colonization by sensitive isolates, but not by any resistant isolates. Four resistant and four sensitive isolates were compared for fitness components and their relative competitive ability on Lupinus Russell Hybrids in the absence of mefenoxam. Resistant isolates outcompeted sensitive ones within 3 to 6 sporulation cycles on lupin seedlings, regardless of their initial proportions in mixed zoospore inoculum. Resistant isolates exhibited greater infection rate and higher sporulation ability than sensitive ones when they were applied separately onto lupins. These results suggest that fungicide resistance may pose a serious challenge to the continued effectiveness of mefenoxam as a control option for nursery growers.     

Synergistic and antagonistic interactions of terpenes against Meloidogyne incognita and the nematicidal activity of essential oils from seven plants indigenous to Greece

BACKGROUND: Biorational means for phytonematode control were studied within the context of an increasingly ecofriendly pest management global approach. The nematicidal activity and the chemical composition of essential oils (EOs) isolated from seven plants grown in Greece and ten selected compounds extracted from them against second‐stage juveniles (J2) of Meloidogyne incognita (Kof. & White) Chitwood were evaluated using juvenile paralysis experiments. Additionally, synergistic and antagonistic interactions between nematicidal terpenes were studied using an effect addition model, with the comparison made at one concentration level. RESULTS: The 96 h EC₅₀ values of Foeniculum vulgare Mill., Pimpinella anisum L., Eucalyptus meliodora A Cunn ex Schauer and Pistacia terebinthus L. were 231, 269, 807 and 1116 µg mL^?1, respectively, in an immersion bioassay. Benzaldehyde (9 µg mL^?1) was the most toxic compound, followed by γ‐eudesmol (50 µg mL^?1) and estragole (180 µg mL^?1), based on 96 h EC₅₀ values. The most potent terpene pairs between which synergistic actions were found, in decreasing order, were: trans‐anethole/geraniol, trans‐anethole/eugenol, carvacrol/eugenol and geraniol/carvacrol. CONCLUSION: This is the first report on the activity of F. vulgare, P. anisum, E. meliodora and P. terebinthus, and additionally on synergistic/antagonistic nematicidal terpene interactions, against M. incognita, providing alternative methods for nematode control. Copyright © 2010 Society of Chemical Industry     

Insecticidal activities of leaf and twig essential oils from Clausena excavata against Aedes aegypti and Aedes albopictus larvae

BACKGROUND: The current study investigates, for the first time, the mosquito larvicidal activities of leaf and twig essential oils from Clausena excavata Burm. f. and their individual constituents against Aedes aegypti L. and Aedes albopictus Skuse larvae. The yields of essential oils obtained from hydrodistillation were compared, and their constituents were determined by GC‐MS analyses. RESULTS: The LC₅₀ values of leaf and twig essential oils against fourth‐instar larvae of Ae. aegypti and Ae. albopictus were 37.1–40.1 µg mL^?1 and 41.1–41.2 µg mL^?1 respectively. This study demonstrated that C. excavata leaf and twig essential oils possess mosquito larvicidal activity, inhibiting the growth of mosquito larvae for both species at a low concentration. In addition, results of larvicidal assays showed that the effective constituents in leaf and twig essential oils were limonene, γ‐terpinene, terpinolene, β‐myrcene, 3‐carene and p‐cymene. The LC₅₀ values of these constituents against both mosquito larvae were below 50 µg mL^?1. Among these effective constituents, limonene had the best mosquito larvicidal activity, with LC₅₀ of 19.4 µg mL^?1 and 15.0 µg mL^?1 against Ae. aegypti and Ae. albopictus larvae respectively. CONCLUSION: The findings suggested that the essential oils from Clausena excavata leaf and twig and their effective constituents may be explored as a potential natural larvicide. Copyright © 2008 Society of Chemical Industry     

Inheritance mode and realized heritability of resistance to imidacloprid in the brown planthopper,Nilaparvata lugens (Stål) (Homoptera: Delphacidae)

BACKGROUND: The brown planthopper, Nilaparvata lugens (Stål), is a serious pest that causes enormous losses to the rice crop in Asia. The genetic basis of imidacloprid resistance was investigated in N. lugens. RESULTS: The resistant strain, selected for imidacloprid resistance from a field population of N. lugens collected from Nanjing, Jiangsu Province, China, showed a 964‐fold resistance compared with the laboratory strain. Progenies of reciprocal crosses (F₁ and F₁′) showed similar dose–mortality responses (LC₅₀) to imidacloprid, and also exhibited a similar degree of dominance (D), 0.58 for F₁ and 0.63 for F₁′. Chi‐square analyses of self‐bred and backcross progenies (F₂, F₂′ and BC respectively) rejected the hypothesis for a single gene control of the resistance. The estimated realized heritability (h²) of imidacloprid resistance was 0.1141 in the resistant strain of N. lugens. CONCLUSION: The results showed that imidacloprid resistance in N. lugens was autosomal and was expressed as an incompletely dominant trait, probably controlled by multiple genes. Copyright © 2009 Society of Chemical Industry     

Identification and characterisation of ten glutathione S-transferase genes from oriental migratory locust, Locusta migratoria manilensis (Meyen)

BACKGROUND: Synthetic pyrethroids are the primary insecticides that are widely used for controlling Locusta migratoria manilensis (Meyen), a major pest in eastern and southern Asia and the Pacific region. In this paper, ten cDNAs encoding glutathione S‐transferases (GSTs) were sequenced and characterised in L. migratoria manilensis. The effects of deltamethrin on the ten GST gene expressions were studied. RESULTS: Phylogenetic analysis revealed nine GSTs in three different classes, including seven in sigma, one in delta and one in theta. The remaining GST (LmGSTu1) was unclassified. RT‐PCR analysis showed that most GST genes were expressed in all tissues examined, including the foregut, midgut, gastric caecum, hindgut, Malpighian tubules, fat bodies, muscles, spermaries and ovaries, except that LmGSTs2, LmGSTs4, LmGSTs7 and LmGSTu1 were expressed in several tissues. LmGSTu1 appeared to be the only gene whose expressions could not be detected in eggs. Real‐time quantitative PCR showed that deltamethrin at 0.08 and/or 0.12 µg mL^?1 increased almost all ten GST gene expressions in third‐instar nymph locusts. However, deltamethrin at 0.16 and/or 0.2 µg mL^?1 decreased the expressions of LmGSTd1, LmGSTs1, LmGSTs5 and LmGSTs6. CONCLUSION: The increases in GST gene expressions after deltamethrin exposure in L. migratoria manilensis might result in its elevating tolerance to other insecticides and xenobiotics. Copyright © 2011 Society of Chemical Industry     

Effects of pymetrozine, an antifeedant of Hemiptera, on Asian citrus psyllid, Diaphorina citri, feeding behavior, survival and transmission of Candidatus Liberibacter asiaticus

BACKGROUND: Vector‐borne plant disease management can be enhanced by deployment of antifeedants in addition to the use of broad‐spectrum neurotoxic insecticides. The effects of pymetrozine on Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, feeding behaviour, survival and transmission of Candidatus Liberibacter asiaticus (Las), the presumed causal pathogen of huanglongbing, were investigated. RESULTS: Pymetrozine applied at 52 and 104 µg mL^?1 to citrus plants [Swingle citrumelo (X Citroncirus webberi Ingram and Moore)] modified the feeding behavior of ACP and increased the amount of time spent performing non‐penetration behaviors while decreasing the time spent performing ingestion behaviors compared with the controls 1 day after treatment. However, the antifeedant effect of pymetrozine subsided 5 days after application. Pymetrozine reduced the survival of both adults and nymphs on treated plants compared with the control. However, it had a greater impact on survival of nymphs than on survival of adults. Pymetrozine applied at 52 and 104 µg mL^?1 on Las‐infected ‘Valencia’ sweet orange plants [Citrus sinensis L. (Osbeck)] reduced acquisition (12 and 21% respectively) and transmission (11 and 18% respectively) of Las by feeding ACP adults compared with the controls; however, these reductions were not statistically significant. CONCLUSIONS: Pymetrozine exhibited moderate antifeedant effects by modifying the feeding behavior of ACP adults with short residual activity. The impact of pymetrozine on survival of nymphs was greater than on adults at the higher concentrations tested. Pymetrozine also reduced the acquisition and transmission of Las by feeding ACP adults up to 21 and 18%, respectively, compared with untreated controls. Copyright © 2010 Society of Chemical Industry     

Inheritance of resistance to Bacillus thuringiensis Cry1Ac toxin in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) from India

BACKGROUND: The cotton bollworm, Helicoverpa armigera (Hübner), is one of the most serious insect pests of cotton. It has developed resistance to almost all groups of chemical insecticides because of their intensive use. The failure of insecticides to control H. armigera has been a strong incentive for the adoption of transgenic cotton (Bt cotton). However, the value of Bt could be diminished by widespread resistance development to Bt toxins in insect populations. Therefore, understanding the genetic basis of resistance is essential for developing and implementing strategies to delay and monitor resistance. RESULTS: A resistant strain designated as BM‐R was obtained from the cross of adults from Bathinda () and Muktsar (), Punjab, India, which showed the highest survival (60.68%) and LC₅₀ value (1.396 µg mL^?1 diet). Similarly, a laboratory‐maintained strain from Hoshiarpur, Punjab, showed maximum susceptibility to Cry1Ac toxin with the lowest LC₅₀ value (0.087 µg mL^?1), and was designated as HP‐S. The genetic purity of both strains was confirmed by RAPD profile analysis at each generation, and genetic similarity reached more than 90% after the third generation. Continuous maintenance of the resistant BM‐R strain on Cry1Ac resulted in an increase in LC₅₀ from 0.531 µg mL^?1 in F₀ to 4.28 µg mL^?1 in F₁₄ and 7.493 µg mL^?1 in F₁₉, while the LC₅₀ values for HP‐S larvae on diet without Cry1Ac increased to 0.106 and 0.104 µg mL^?1, which lay within the fiducial limits of the baseline LC₅₀ value. The mode of inheritance of resistance was elucidated through bioassay response of resistant, susceptible heterozygotes and backcross progeny to Cry1Ac incorporated in semi‐synthetic diet. CONCLUSION: Based on dominance, degree of dominance and backcross values, resistance was inferred to be polygenic, autosomal and inherited as a recessive trait. Copyright © 2011 Society of Chemical Industry     

Assessment of the high-dose concept and level of control provided by MON 87701 × MON 89788 soybean against Anticarsia gemmatalis and Pseudoplusia includens (Lepidoptera: Noctuidae) in Brazil

BACKGROUND: Genetically modified MON 87701 × MON 89788 soybean (Glycine max), which expresses the Cry1Ac and EPSP‐synthase proteins, has been registered for commercial use in Brazil. To develop an Insect Resistance Management (IRM) program for this event, laboratory and field studies were conducted to assess the high‐dose concept and level of control it provides against Anticarsia gemmatalis and Pseudoplusia includens. RESULTS: The purified Cry1Ac protein was more active against A. gemmatalis [LC₅₀ (FL 95%) = 0.23 (0.15–0.34) µg Cry1Ac mL^?1 diet] than P. includens [LC₅₀ (FL 95%) = 3.72 (2.65–4.86) µg Cry1Ac mL^?1 diet]. In bioassays with freeze‐dried MON 87701 × MON 89788 soybean tissue diluted 25 times in an artificial diet, there was 100% mortality of A. gemmatalis and up to 95.79% mortality for P. includens. In leaf‐disc bioassays and under conditions of high artificial infestation in the greenhouse and natural infestation in the field, MON 87701 × MON 89788 soybean showed a high level of efficacy against both target pests. CONCLUSIONS: The MON 87701 × MON 89788 soybean provides a high level of control against A. gemmatalis and P. includes, but a high‐dose event only to A. gemmatalis. Copyright © 2012 Society of Chemical Industry     

Influence of the nitrogen form on in vitro organogenesis in Equisetum arvense

The physiological and biochemical mechanisms of organogenesis in Equisetum arvense have not been clarified yet. However, high concentrations of nitrogen have been shown to exert an inhibitory effect on in vitro tuber formation in E. arvense. The aim of this study was to clarify the influence of the form of nitrogen in a medium on in vitro organogenesis in E. arvense. Single‐node segments of E. arvense rhizomes were cultured in the test medium. The NH₄‐N and NO₃‐N concentrations of the test medium, respectively, were adjusted by adding NH₄H₂PO₄ and KNO₃ to the basal medium. The basal medium was a nitrogen‐free, modified form of White's medium. Vegetative shoots were newly formed in the test tubes for concentrations of NO₃‐N and NH₄‐N that exceeded 56 mg L^?1. However, no rhizome was formed at NH₄‐N concentrations exceeding 28 mg L^?1. The number of newly formed tubers decreased at an NH₄‐N concentration of 28 mg L^?1 and no tuber was formed at NH₄‐N concentrations exceeding 56 mg L^?1. In summary, although the presence of NO₃‐N in the medium did not inhibit in vitro rhizome or tuber formation in E. arvense, the presence of NH₄‐N in the medium exerted a strong inhibitory effect on the in vitro formation of both of these organs.     

Sensitivity of Botrytis cinerea to chitosan and acibenzolar‐S‐methyl

BACKGROUND: The antifungal properties of chitosan and acibenzolar‐S‐methyl were evaluated to assess their potential for protecting grapes against Botrytis cinerea Pers.: Fr. isolated from Vitis vinifera L. The objectives were to determine the effects of these compounds on the in vitro development of B. cinerea and to assess their effectiveness at controlling grey mould on grapes stored at different temperatures. RESULTS: Both agents significantly inhibited the radial growth of this fungus species. The EC₅₀ was 1.77 mg mL^?1 for chitosan and 3.44 mg mL^?1 for acibenzolar‐S‐methyl. In addition, single grapes treated with aqueous solutions of chitosan (1.0 and 2.5 mg mL^?1) and acibenzolar‐S‐methyl (1.0 and 3.0 mg mL^?1) were inoculated with B. cinerea and incubated at both 4 and 24 °C. After 4 days at 24 °C, all the concentrations of chitosan and acibenzolar‐S‐methyl significantly reduced B. cinerea growth. However, at 4 °C, significant differences were only observed between chitosan at 2.5 mg mL^?1 and acibenzolar‐S‐methyl at both 1.0 and 3.0 mg mL^?1 and the corresponding controls. After 3 days at 24 °C, the greatest reduction in lesion size was obtained in grapes pretreated with acibenzolar‐S‐methyl at 3.0 mg mL^?1. Only the highest doses of these products significantly reduced the lesion diameters when grapes were stored for 3 days at 4 °C. CONCLUSIONS: Chitosan and acibenzolar‐S‐methyl could directly inhibit the growth of Botrytis cinerea in vitro and confer resistance on grapes against grey mould. Pretreatment with these compounds could be an alternative to traditional fungicides in post‐harvest disease control in grapes. Copyright © 2010 Society of Chemical Industry