Chloroplast DNA molecular characterization and leaf volatiles analysis of mint (Mentha; Lamiaceae) populations in China

Non-coding chloroplast DNA (cpDNA) was used to evaluate genetic diversity and relationships within the section Mentha collected from China. Leaf volatiles of Mentha spicata accessions and its parent were analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). To this end, genetic diversity of 30 Mentha accessions from different geographic origins of China, representing six species and two hybrids, was assessed. Molecular studies grouped the samples mainly according to genetic relationship established by conventional methods. Cluster analysis based on the leaf volatiles chemical composition of M. spicata accessions defined two main chemotypes: 1,8-cineole-piperitenone oxide type and limonene-carvone type. The results suggested that there was a high genetic variability among individuals of Mentha in China. A distinct correlation between cpDNA marker and volatile oils in M. spicata accessions was revealed. Moreover, the chemical composition followed maternal inheritance in M. spicata accessions. In conclusion, the genetic diversity of Mentha populations as shown in this study should play a critical role in future selection and breeding programs.     

Hybanthus parviflorus (Violaceae): Insecticidal activity of a South American plant

Hybanthus parviflorus is a perennial shrub, widely distributed in the tropical and subtropical regions of America, and is known as ‘violetilla’ in Argentina. Previous phytochemical studies of this species led us to the isolation and determination of the primary structure of a novel macrocyclic polypeptide, the cyclotide hypa A. Here the insecticidal activities of extracts of H. parviflorus were determined against Ceratitis capitata Wied., the Mediterranean fruit fly or ‘Medfly’. Mortality in the different life stages of Medfly, total mortality and modifications of the insect’s physiology caused by 50% EtOH and CH₂Cl₂ extracts of H. parviflorus were evaluated. In addition, we determined the occurrence of ursolic acid, β-sitosterol and the polyphenols quercetin, quercetin-3-methyl ether, apigenin, luteolin, kaempferol, rutin, caffeic acid and chlorogenic acid. The promising insecticidal activity of 50% EtOH extracts of H. parviflorus and its purified fractions was related to the presence of cyclotides. The insecticidal activity of CH₂Cl₂ extracts could be related to the presence of polyphenols, ursolic acid and β-sitosterol.     

Insecticidal activities of crude extracts and phospholipids from Chenopodium ficifolium against melon and cotton aphid, Aphis gossypii

Several organic solvent extracts of Chenopodium ficifolium were tested for their insecticidal activity against melon and cotton aphid, Aphis gossypii, on cucumber plants. Both methanol and ethanol extracts, at 5000 μg ml⁻¹, were highly active giving over 80% control. The other crude extracts displayed moderate or weak insecticidal activity giving control in the range of 16–69%. Two phospholipids were isolated as insecticidal active substances from C. ficifolium. Their chemical structures were identified as 1-palmitoyl-2-(3-trans)-hexadecenoyl-sn-glycero-3-phosphoglycerol and 1-palmitoyl-2-linoleoyl-3-glycerophosphocholine by GC–MS, EDS, mass and NMR spectral analyses. Both compounds displayed a dose-dependent mortality of A. gossypii. Furthermore, the liquid formulation that was obtained by partitioning with n-hexane from the methanol extract of C. ficifolium controlled melon and cotton aphid on cucumber plants effectively. These results indicate that extracts of C. ficifolium have potential for development as botanical insecticides for controlling A. gossypii infesting cucumber plants.     

Insecticidal formulation based on Xylopia aethiopica essential oil and kaolinite clay for maize protection

Insecticidal formulations based on the essential oil of Xylopia aethiopica (Annonaceae) and kaolinite-clay (particles size less than 50 μm) are efficient in protecting stored grains. Ingestion-contact insecticide tests have been conducted using the maize weevil Sitophilus zeamais (Motsch.) (Curculionidae). Residual action of terpenic constituents adsorbed on clay was assessed. Bioassays indicated that S. zeamais adult mortality increased with the quantity of active ingredient. The formulation based on essential oil and clay produced mortality rates ranging from 22% at 2.5% (w/w) to 100% at 10% (w/w). The lethal concentration producing a 50% mortality rate (LC50) was 4.35% (w/w). Efficacy also depended on storage conditions and duration of the formulation, decreasing from 91 to 37% for formulation stored in closed boxes and from 91 to 17% for formulation stored in open boxes, after 1 and 15 days respectively.     

Comparative analysis of essential oil components of two Cryptomeria species from China

The essential oils obtained by hydrodistillation from leaves of Chinese native Cryptomeria japonica and Cryptomeria fortunei were analyzed by GC and GC-MS. Fifty-seven compounds were identified in the essential oils of C. japonica with α-elemol (20.12%), kaur-16-ene (14.84%), β-phellandrene (5.97%), β-elemene (5.87%), α-eudesmol (5.62%) and β-eudesmol (5.03%) as main constituents. Forty components were identified in the oil of C. fortunei with kaurene (34.04%), α-elemol (13.34%), γ-eudesmol (10.80%), β-eudesmol (10.16%), α-pinene (2.75%) and γ-cadinene (1.92%) as the most abundant components. This study demonstrated the occurrence of α-elemol chemotype in C. japonica and kaurene chemotype in C. fortunei from China. The essential oil compositions of two Cryptomeria samples were shown that they can be used for green plant protection, pharmaceutical, perfume and food industries.     

Insecticidal effects of Ungernia severtzovii bulb extracts against the grain aphid Schizaphis graminum (Rondani)

Insecticidal effect of Ungernia severtzovii bulbs extract has been investigated against the grain aphid Schizaphis graminum. LC₅₀ value of the ethanolic extract of U. severtzovii against the female aphids was 0.235% (2.35 g/l). There was 100% mortality of the subsequent nymphs of the treated females at 0.25-1.0%. The extract was equally active against the nymphs at the highest concentrations. Hexane and ethanolic extracts were the most active against the aphids of all the extracts. Fractionation of the hexane extract by TLC yielded three fractions. Fraction II was the most active (>90% mortality) against the aphids. Xanthoxylin was identified as the major constituent in fraction II of the hexane extract and may be responsible for the insecticidal effect of U. severtzovii.     

Insecticidal and repellent activity of selected essential oils against of the pollen beetle, Meligethes aeneus (Fabricius) adults

The essential oils from 9 aromatic plants were tested on repellency and mortality of Meligethes aeneus adults. All the tested essential oils caused high mortality of M. aeneus adults in the tarsal tests. The lethal doses after 6 h exposure were ranged between 197 and 1508 μg cm⁻². Essential oils obtained from Carum carvi and Thymus vulgaris were most efficient where LD₅₀ was estimated as 197 and 250 μg cm⁻², respectively.Repellency declined in all the essential oils as a function of time. The longest persistence time was determined for essences obtained from C. carvi and T. vulgaris where significantly the highest repellent index of 65.6% and 63.8%, respectively, was determined. Repellent index lower than 15% was determined for the remaining essential oils.     

Repellency of selected biorational insecticides to potato psyllid, Bactericera cockerelli (Hemiptera: Psyllidae)

Bactericera cockerelli has recently become a major concern because of its direct feeding and vectoring of bacterial diseases in many solanaceous crops. The repellency of four biorational insecticides, MOI-201 (a Chinese medicine plant extract), Requiem (a plant extract of Chenopodium ambrosioides), BugOil (a mixture of four plant essential oils), and SunSpray oil (a mineral oil), to B. cockerelli adults was tested on tomato. In a no-choice test, all the insecticides had significant repellency to adults and deterred oviposition as compared with untreated controls. Of the four insecticides, the two oils showed a stronger repellency to adults and deterred oviposition more strongly than Requiem or MOI-201. In a choice test, all insecticides had significant repellency to adults and deterred oviposition compared to untreated controls. Of the four tested insecticides, <1 adults and no eggs were found on the leaves treated with SunSpray Oil, BugOil or Requiem 3 d after treatment. The repellency rates of these three insecticides were 77.2–95.4%. MOI-201 also repelled adults significantly and deterred oviposition compared to untreated controls even though it was the least effective insecticide among the four evaluated. In conclusion, all four insecticides tested showed significant repellency to B. cockerelli adults and deterred oviposition, especially the two oils. The overall repellency to potato psyllid adults can be arranged in a descending order of SunSpray oil > BugOil > Requiem > MOI-201. These insecticides could be used in integrated pest management programs targeted against the potato psyllid on solanaceous crops.     

Chemical variation in essential oil of Cryptomeria fortunei from various areas of China

Essential oils were extracted by hydrodistillation from the leaves of Cryptomeria fortunei from the principal habitats in China. The essential oil yields of samples from various locations ranged from 0.61 to 1.66% (w/w). GC-MS analysis revealed eighty-seven compounds, constituting 85.89-94.75% of the total essential oils. The main constituents were kaurene (0-44.69%), phyllocladene (0.07-32.98%), kaur-16-ene (0-32.28%), elemol (5.28-18.92%), α-cadinol (0-11.39%), α-eudesmol (0-11.21%), α-pinene (2.32-9.15%). The samples were separated into 3 chemotypes by hierarchical cluster analysis, according to the composition of the corresponding essential oils. Chemical composition of essential oils was significantly related to nine geographic distributions among populations. Our results revealed chemical polymorphism in the essential oils of the C. fortunei leaf in China. The chemical variability could be attributed to genetic and environmental factors. The chemical diversity of the oils offers opportunity to choose C. fortunei leaves and essential oils with preferential compound for green plant protection, pharmaceutical, perfume and food industries.     

Insecticidal effect of recombinant endophytic bacterium containing Pinellia ternata agglutinin against white backed planthopper, Sogatella furcifera

White backed planthopper (WBPH; Sogota furcifera Horvath) has become the major threat to rice crops throughout Asia, damaging plants both through its feeding behavior and by acting as a virus vector. Here, we developed a novel method for biologically controlling WBPH by using endophytic bacterium to express anti-pest plant lectins. Strain SJ-10 of an endophytic bacterium, characterized as Enterobacter cloacae by morphological, physiological, biochemical and 16s rDNA characteristics, was isolated from rice seedlings. The Pinellia ternate agglutinin (PTA) gene was cloned into SJ-10 for expression. The positive transformant, selected by antibiotic resistance, was evaluated using PCR, SDS-PAGE and Western blot assay. After inoculation, rSJ-10 could colonize rice plants so that they expressed PTA, and then the rice was shown to have insecticidal activity against WBPH. The results showed that rSJ-10 could significantly decrease the survival and fecundity of WBPH fed on rice seedlings (p < 0.01). At day 19, the fecundity of WBPH inoculated with rSJ-10, or with wild-type SJ-10 was decreased by 86.1%, and 25.6%, respectively. At day 22, numbers of WBPH on rice in the control were 19.4 times greater than on rice inoculated with rSJ-10. At day 26, the rice seedlings all died in the control group, but the seedlings inoculated with rSJ-10 grew well. The results showed that the rice seedlings inoculated with rSJ-10 expressing PTA protein were endowed with the anti-pest activity against WBPH. Further work is needed to investigate whether the rice plants expressing rPTA are toxic to mammals. This research highlights a way to biologically control planthoppers by recombinant endophytic bacteria expressing plant lectins.     

High level of resistance to Sclerotinia sclerotiorum in introgression lines derived from hybridization between wild crucifers and the crop Brassica species B. napus and B. juncea

Sclerotinia rot caused by the fungus Sclerotinia sclerotiorum is one of the most serious and damaging diseases of oilseed rape and there is keen worldwide interest to identify Brassica genotypes with resistance to this pathogen. Complete resistance against this pathogen has not been reported in the field, with only partial resistance being observed in some Brassica genotypes. Introgression lines were developed following hybridization of three wild crucifers (viz. Erucastrum cardaminoides, Diplotaxis tenuisiliqua and E. abyssinicum) with B. napus or B. juncea. Their resistance responses were characterized by using a stem inoculation test. Seed of 54 lines of B. napus and B. juncea obtained from Australia, India and China through an Australian Centre for International Agricultural Research (ACIAR) collaboration programme were used as susceptible check comparisons. Introgression lines derived from E. cardaminoides, D. tenuisiliqua and E. abyssinicum had much higher levels (P < 0.001) of resistance compared with the ACIAR germplasm. Median values of stem lesion length of introgression lines derived from the wild species were 1.2, 1.7 and 2.0 cm, respectively, as compared with the ACIAR germplasm where the median value for stem lesion length was 8.7 cm. This is the first report of high levels of resistance against S. sclerotiorum in introgression lines derived from E. cardaminoides, D. tenuisiliqua and E. abyssinicum. The novel sources of resistance identified in this study are a highly valuable resource that can be used in oilseed Brassica breeding programmes to enhance resistance in future B. napus and B. juncea cultivars against Sclerotinia stem rot.     

Identification of an insecticidal polyacetylene derivative from Chrysanthemum macrotum leaves

Compounds responsible for insecticidal properties of Chrysanthemum macrotum (D.R.) Ball. leaves against Spodoptera littoralis Boiduval caterpillars have been investigated. The screening of the insecticidal activity was performed by incorporating methanol, buthanol or ethyl acetate extracts, or some chromatographic fractions to the caterpillars’ artificial diet. It was noted that extracts and fractions ameliorated or disturbed nutritional indexes, being not always toxic for caterpillars. Among the tested fractions, one pure compound with a high insecticidal activity (percentage of mortality 66.7%) was purified. The nuclear magnetic resonance study allowed its identification as a polyacetylene derivative, in particular a spiroketal enol ether one.     

A comparison of monitoring systems used for Ceratitis species (Diptera: Tephritidae) in South Africa

For 16 years a bucket-type trap known as the Sensus fruit fly trap has been used to monitor three fruit fly pest species Ceratitis capitata, Ceratitis rosa and Ceratitis cosyra in South African fruit industries. The relative efficiencies of lures sold for monitoring fruit flies with the Sensus trap in South Africa were determined in field experiments where laboratory-reared C. capitata, C. rosa and C. cosyra were released in a mango orchard within a few metres of Sensus traps containing either Capilure (trimedlure or tert-butyl 4 (or 5) -chloro-2-methylcyclohexane carboxylate), Ceratitislure (protein hydrolysate plus β-caryophyllene) or Questlure (protein hydrolysate plus plant extracts). When using 12-day-old flies, Capilure caught 3 times more C. capitata males than C. rosa males and this difference was more extreme when 3-day-old flies were released. Ceratitislure caught significantly more 12-day-old C. cosyra males than 12-day-old C. capitata males, but the difference was reversed when 3-day-old flies were compared. Questlure showed the least differences between species and age but recovered the lowest proportion of released species. Further comparisons were conducted in an orchard with wild flies using other known attractants in larger yellow Probodelt bucket traps. Capilure caught more male C. capitata than BioLure Fruit Fly 3-component, but BioLure 2-component (trimethylamine and ammonium acetate) was more effective than Questlure for C. capitata females. The 3-component lure was also more effective than both Capilure and Questlure for male and female C. rosa, respectively. Ceratitislure was the most effective lure for male C. cosyra flies and BioLure 3-component was more effective than Ceratitislure and Questlure for female C. cosyra flies. The intervention threshold of 4 flies/trap/week previously used in citrus with Capilure for C. rosa was lowered to 2 C. rosa/trap/week when using the Sensus trap due to the lower sensitivity of this trap-lure combination found for C. rosa in this study. The 3-component lure, or the 2-component combination of trimethylamine and ammonium acetate in a ratio of 1:8 in the Sensus trap capsule, would be more effective for both sexes of all three Ceratitis species than the Questlure that is currently being used.     

Comparative susceptibility of Ostrinia furnacalis, Ostrinia nubilalis and Diatraea saccharalis (Lepidoptera: Crambidae) to Bacillus thuringiensis Cry1 toxins

Transgenic corn hybrids that express toxins from Bacillus thuringiensis (Bt) are highly effective against the European corn borer, Ostrinia nubilalis (Hübner), and the closely related Asian corn borer, Ostrinia furnacalis (Guenée). Since the registration of Bt corn hybrids in the U.S. in 1996, there has been a great deal of information generated on O. nubilalis. However, relatively little information exists for O. furnacalis. To help determine whether the information generated for O. nubilalis can be leveraged for decisions regarding the use of transgenic Bt corn against O. furnacalis, experiments were designed to determine whether the pattern of sensitivity to various Bt Cry1 toxins is similar between the two species. Test insects included laboratory-reared O. furnacalis originating from Malaysia, a Bt-susceptible laboratory colony of O. nubilalis maintained at the University of Nebraska-Lincoln (UNL) and an out-group consisting of the sugarcane borer, Diatraea saccharalis (F.), from Louisiana which represents a different genus from the same family. O. furnacalis and O. nubilalis exhibited a similar pattern of susceptibility to all the Cry1 toxins and were highly susceptible to the range of Bt toxins tested including Cry1Aa, Cry1Ab, Cry1Ac and Cry1F. Both of the Ostrinia species were more tolerant to Cry1Ba compared with D. saccharalis, although sensitivity of O. furnacalis was intermediate and did not differ significantly from that of O. nubilalis and D. saccharalis. D. saccharalis was also susceptible to the range of toxins tested but unlike the two Ostrinia species, was more tolerant to Cry1F and more susceptible to Cry1Ba. These results indicate that both of the Ostrinia corn borer species are similar in sensitivity to the Cry1Aa, Cry1Ab, Cry1Ac, Cry1Ba and Cry1F toxins, thus suggesting shared toxin receptors and mechanisms of toxicity for the two species.     

Comparative overwintering host range of three Adelphocoris species (Hemiptera: Miridae) in northern China

In northern China, Adelphocoris suturalis, Adelphocoris lineolatus and Adelphocoris fasciaticollis (Hemiptera: Miridae) are common pests of cotton and several other crops. These species have vastly diverse geographic distribution, seasonal dynamics and abundance, the underlying causal factors of which are poorly understood. In this study, the importance of a broad range of plant species as overwintering hosts for each Adelphocoris sp. was compared. Nymphal emergence from a total of 126 plant species was monitored at two distinct locations. The eggs of A. suturalis successfully eclosed from un-plowed cotton field soil and 115 plant species, primarily pastures, weeds and agricultural crops. The eggs of A. lineolatus successfully eclosed from 40 plant species, mainly pastures and weeds. Finally, A. fasciaticollis overwintered on 35 plant species, primarily tree species, weeds and agricultural crops. In conclusion, the most common and widely distributed mirid species, A. suturalis, overwintered on a comparatively broader range of plants compared to the other two species. These observations help to understand the differences in geographical distribution and abundance of the three Adelphocoris species, and constitute the basis for forecasting and pest management protocols for Adelphocoris spp. in China.     

A comparative view of grain development in Brachypodium distachyon

Cereal grains are both at the forefront of agriculture as a food source and in basic botanical terms are unique fruit forms. Analyses and characterisation of their development and composition are therefore invaluable in both applied and basic biological research. A key approach in driving forward this research is a comparative one and this approach is facilitated when there are sequence, gene expression and functional resources available for a variety of species. In this article we review and assess the current status of resources for Brachypodium distachyon as a model system and we then focus specifically on recent studies characterising and comparing grain development, organisation and composition in this species.     

Composition of essential oil, concrete, absolute and SPME analysis of Tagetes patula capitula

The chemical compositions of essential oil, concrete, absolute from the capitula of Tagetes patula (family Asteraceae) were analysed by GC-FID and GC/MS. The major compounds identified were (Z)-β-ocimene, (E)-β-ocimene, terpinolene, (Z)-ocimenone, (E)-ocimenone and δ-elemene. In addition, the volatiles of live and plucked capitula (flowers) were analysed by SPME technique using PDMS/DVB/CAR fiber. The SPME-GC-FID analyses of live capitula showed that the volatiles were rich in monoterpenoids in comparison to the plucked capitula. On the other hand, the plucked capitula recorded with significant increase in sesquiterpenoids in comparison to the living capitula.     

Population dynamics of Xanthomonas campestris pv. vitians on different plant species and management of bacterial leaf spot of lettuce under greenhouse conditions

The population dynamics of Xanthomonas campestris pv. vitians (Xcv) was studied both externally and internally in lettuce, tomato and pepper plants. In addition, the application of bactericides during transplant production period was carried out for the management of bacterial leaf spot of lettuce under greenhouse conditions. Epiphytic populations of Xcv were recovered on leaves of lettuce plants (10⁵ CFU/g) 5 weeks after sprayed than the other plant species when inoculated with 10⁸ CFU/ml of Xcv. When plants of each crop species infiltrated with the bacterium at 10⁵ CFU/ml, the highest populations were developed in lettuce (10⁸ CFU/cm²) followed by pepper with 10⁶ CFU/cm² and tomato with 10⁵ CFU/cm² 10-days after infiltration. Application of a mixture of Maneb and Kocide or Kocide alone as a foliar spray on lettuce significantly reduced the incidence and disease severity of bacterial leaf spot by 29 and 27% respectively. Spread of the bacterium and development of the disease may partly be managed by avoiding intercropping of these plants commonly grown in close proximity to lettuce. For the management of leaf-associated populations of Xcv in lettuce, use of a mixture of Maneb and Kocide is advocated to minimize the effect of attacks.     

Improvement of essential oil yield of oil-bearing (Rosa damascena Mill.) due to surfactant and maceration

The essential oil content of the oil-bearing rose (Rosa damascene Mill.) is relatively low, around 0.3-0.4 mL kg⁻¹ in fresh flowers. There is a need to increase essential oil yield of oil-bearing rose. The objective was to examine the effect of Tween 20 (polyoxyethylene sorbitan monolaurate) applied with, or without, maceration of flowers on oil content and composition of oil-bearing rose harvested at beginning of flowering, full bloom, and end of flowering. Addition of Tween 20 at 1000 mL L⁻¹ and 2500 mL L⁻¹ increased essential oil yield by 26% (to 0.44 mL kg⁻¹) and 54% (to 0.54 mL kg⁻¹) respectively relative to the untreated control that gave 0.35 mL kg⁻¹ yield. Maceration, in combination with the addition of Tween 20 at 1000 mL L⁻¹ and Tween 20 at 2500 mL L⁻¹, increased oil yield by 69% (to 0.59 mL kg⁻¹) and 94% (to 0.68 mL kg⁻¹) respectively. Among the three phenological phases of harvest, harvesting at the beginning of flowering gave the highest yield followed by the full bloom and then by the end of flowering phases. Since the interaction effect was not significant, the differences obtained among the treatments were regardless of the phase, and vice versa. Treatments did not significantly alter composition of the essential oil. Postharvest pre-extraction application of Tween 20 in combination with maceration could be used in the rose industry for increasing the essential oil yield.     

Chemical composition and antioxidant and antimicrobial activity of essential oil of Artemisia annua L. from Bosnia

Hydrodistilled volatile oil obtained from the aerial parts of Artemisia annua L., cultivated near Sarajevo, Bosnia, was analyzed by GC-MS. More than one hundred compounds were identi?ed, representing 95.5% of the total oil. The major constituents of essential oil were oxygenated monoterpenes, artemisia ketone (30.7%) and camphor (15.8%). Isolated essential oil was tested for radical-scavenging ability using the stable DPPH radical, the ABTS radical, for reducing power ability with a test based on the reduction of ferric cations, for reducing ability of hydroxy radical in ORAC assay, and for metal chelating ability using the ferrozine assay. In all tests oil did not show a prominent antioxidant activity, but still comparable with thymol, an already known antioxidant. The screening of antimicrobial activity of oil was individually evaluated against representatives of Gram-positive, Gram-negative bacteria and fungi, using the agar diffusion method. All tested microorganisms were inhibited by essential oil. To the best of our knowledge, this is the first report of antimicrobial activity of essential oil of A. annua against Haemophilus influenzae, Enterococcus faecalis, Streptococcus pneumoniae, Micrococcus luteus and Candida krusei microbial strains. The antioxidant, antibacterial and antifungal activity of essential oil of A. annua from Bosnia is presented here for the first time and extends our knowledge in the range of valuable biological activities and possible roles in therapy associated with this medicinal herb.