Exotic wildland weeds serve as reservoirs for a newly introduced cole crop pest,Bagrada hilaris (Hemiptera: Pentatomidae)

The role of wildland weeds that serve as alternate hosts for insect pests has not been directly examined for the potential to sustain pest populations or facilitate pest outbreaks. The development of weed biological control programmes is also complicated by weed species that are closely related to economically important or native plants, especially rare or special status species. In recent surveys in southern California, USA, we found a newly introduced insect pest of cole crops, Bagrada hilaris Burmeister (Bagrada bug), feeding on Brassicaceae weeds in riparian areas adjacent to agricultural fields where cole crops are routinely grown. Insect populations grew to levels well over action thresholds and caused severe damage to populations of the invasive weed, Lepidium latifolium (perennial pepperweed). The numerical response of B. hilaris on L. latifolium and other Brassicaceae weeds in natural areas may pose a significant challenge to effectively managing pest populations in crops. However, the accidental introduction of this insect provides the opportunity to examine plant–insect interactions with important implications for development of biologically based control methods for weeds.     

Root damage to apple plants by cockchafer larvae induces a change in volatile signals below‐ and above‐ground

Volatile organic compounds (VOCs) mediate communication between plants and insects. Plants under insect herbivore attack release VOCs either at the site of attack or systemically, indicating within‐plant communication. Some of these VOCs, which may be induced only upon herbivore attack, recruit parasitoids and predatory insects to feed on the attacking insects. Moreover, some plants are able to ‘eavesdrop’ on herbivore‐induced plant volatiles (HIPVs) to prime themselves against impending attack; such eavesdropping exemplifies plant–plant communication. In apple orchards, the beetle Melolontha melolontha L. (Coleoptera: Scarabaeidae) is an important insect pest whose larvae live and feed on roots for about 4 years. In this study, we investigated whether the feeding activity of M. melolontha larvae (1) alters the volatile profile of apple roots, (2) induces the release of HIPVs systemically in the leaves, and (3) whether infested plants communicate to neighbouring non‐infested conspecifics through HIPVs. To answer these questions, we collected constitutive VOCs from intact M9 roots as well as M. melolontha larvae‐damaged roots using a newly designed ‘rhizobox’, to collect root‐released volatiles in situ, without damaging the plant root system. We also collected VOCs from the leaf‐bearing shoots of M9 whose roots were under attack by M. melolontha larvae and from shoots of neighbouring non‐infested conspecifics. Gas chromatography‐mass spectrometry analysis showed that feeding activity of M. melolontha larvae induces the release of specific HIPVs; for instance, camphor was found in the roots only after larvae caused root damage. Melolontha melolontha also induced the systemic release of methyl salicylate and (E,E)‐α‐farnesene from the leaf‐bearing shoots. Methyl salicylate and (E,E)‐α‐farnesene were also released by the shoots of non‐infested neighbouring conspecifics. These phenomena indicate the induction of specific VOCs below‐ and above‐ground upon M. melolontha larvae feeding on apple roots as well as plant–plant communication in apple plants.     

Attraction of the potential biocontrol agent Altica cyanea by volatile compounds of three species of Ludwigia weeds from rice fields

Ludwigia adscendens (L.) Hara, Ludwigia parviflora Roxb., and Ludwigia octovalvis (Jacq.) Raven (Onagraceae) are abundant weeds in rice fields in India. These weeds compete with rice for resources in fields and this results in reduction of grain yield. Altica cyanea (Weber) (Coleoptera: Chrysomelidae) is a biocontrol agent of the three rice-field weeds. Hence, it is relevant to study host preference of A. cyanea using volatile cues of these three weeds. Therefore, we attempted to identify volatiles from leaves of the three Ludwigia species attracting A. cyanea, which could be used as an attractant during early emergence of the weeds in rice fields. In Y-tube olfactometer assays, A. cyanea females were more attracted to natural volatiles of plants after 48 h of feeding by adults than to volatiles of undamaged plants. The volatile organic compounds from undamaged plants, and plants after 6 and 48 h of feeding by A. cyanea were identified and quantified by gas chromatography and mass spectrometry (GC–MS) and GC-flame ionization detection (FID), respectively. In total, 25, 29, and 29 volatile compounds were detected in headspaces of undamaged L. adscendens, L. parviflora, and L. octovalvis, respectively, whereas 32, 35, and 34 compounds, respectively, were detected after 48 h of feeding by A. cyanea. Methyl jasmonate predominated among the volatile compounds in all treatments, but this compound was not attractive to A. cyanea. Females were attracted by synthetic blends of 3-hexanol, α-pinene, linalool oxide, and phytol in amounts mimicking those in each of the three Ludwigia species after 48 h of feeding by A. cyanea. The blends mimicking L. adscendens and L. parviflora included geraniol, whereas the blend mimicking L. parviflora also included 1-tridecanol. These synthetic blends may be helpful to monitor A. cyanea in biocontrol programmes.     

Elevated CO2-mediated plant VOCs change aggravates invasive thrips occurrence by altering their host-selection behaviour

Globally rising atmosphere CO₂ has been predicted to affect the emission of plant volatile organic compounds (VOCs) and the interaction with insect herbivores. CO₂-mediated plant VOCs change might contribute to develop effective management strategies for insect pests by using VOCs related biological control methods. In this study, we analysed the effect of elevated CO₂ on the host-selection behaviour of western flower thrips (Frankliniella occidentalis) and studied how they were affected by the release of VOCs from kidney bean plants (Phaseolus vulgaris). A four-chamber olfactometer was used to quantify the host-selection of F. occidentalis for P. vulgaris. Elevated CO₂ increased F. occidentalis' selection for P. vulgaris wounded by mechanical damage (MD) and thrips infestation (TI) that might via regulating the gene expression of CSP1, CSP1-q, OBP1 and OBP1-q in F. occidentalis. Besides, we cultivated kidney beans at ambient CO₂ (400 ppm) and elevated CO₂ (800 ppm), and quantified the emission of plant VOCs by using GC-MS. Thirty VOCs belonging to ten chemical groups were identified from P. vulgaris, including aromatic hydrocarbons, ethers, alkanes, cycloalkanes, alcohols, alkenes, aromatic derivatives, phenols, ketones and esters. Furthermore, six VOCs from P. vulgaris were significantly affected by CO₂ level, wounding way and the interaction between them, including ethyl benzene, 1,3-dimethyl benzene, 1,3-dimethyl-4-ethyl benzene, (E,E,E,E)-squalene, 2,6-ditert-butyl-4-methyl phenol and dioctyl phthalate. Our study indicates that elevated CO₂ might increase the host-selection of F. occidentalis for wounded P. vulgaris due to the changed plant VOCs.     

The potential of Commelina benghalensis as a forage for ruminants

To evaluate the potential of Commelina benghalensis as a forage for ruminants, effects of plant maturity on chemical composition, rumen degradability as well as its increased dietary inclusion level on intake, digestibility and N balance in sheep fed Sorghum almum were investigated with forage obtained from the wild, re-established and harvested at 6, 10 and 14 weeks of growth. Composite herbage samples were analyzed for dry matter (DM), chemical components, total extractable phenolics (TEP) and amino acid content. In sacco rumen degradability measurements used six individually confined wethers (8 ± 0.5 months of age; 21 ± 2.6 kg live-weight (LW)) fitted with rumen cannulae and fed a ration of 3:1 fresh S. almum and Medicago sativa hay (about 1:2 on a DM basis). In sacco bags containing 5 g each of dry herbage were inserted into the rumen and withdrawn sequentially after 0, 12, 24, 36 and 48 h. Metabolizable energy (ME) was estimated from 24 h in vitro gas production. In the digestibility study, 12 wethers fitted with rumen canula were housed in metabolic crates and allotted to four treatment diets (i. e., D0, D10, D20 and D30) constituted from fresh S. almum and pre-wilted C. benghalensis in a randomized complete block design. The control diet (D0) was 3 kg fresh S. almum (≈535.5 g DM/wether/d about 30 g/kg LW), whereas D10, D20 and D30 were D0 +300, 600 or 900 g of wilted C. benghalensis (≈34, 68 or about 102 g DM/wether/d), respectively. The study lasted for 21 d. Dry matter, fibre and TEP content increased (P<0.001) with maturity of the forage, whereas those of CP and EE decreased (P<0.0001) over the same period. Amino acids (AA) also declined with maturity (P<0.05). Rumen degradability of DM and OM were unaffected, but DM intake increased linearly (P<0.0001) at a decreasing rate (Q: P<0.05) and DM digestibility (DMD) and N intake increased linearly (P<0.01 and P<0.0001, respectively) as level of C. benghalensis in the diet increased. Results indicate that advancing maturity affected chemical composition, but not rumen degradability, of C. benghalensis and also demonstrated that inclusion of C. benghalensis in S. almum diet improved intake, digestibility and N intake, suggesting its potential use as a feed supplement.     

Volatiles from Marina neglecta: Biocide effect on insect vectors of tropical diseases in Southern Mexico

Preventive measures based in the control of insect vectors are considered as the best choice to decrease the incidence of insect-borne diseases. Herein we report on the volatile content of the leaf essential oils from Marina neglecta, a medicinal plant distributed in the tropical regions of southern Mexico. In order to investigate the chemical variation of the essential oils, a volatile screening was performed during the four seasons of the years 2016–2019. Simultaneously, their biological activity was tested on distinct life stages of Meccus pallidipennis, M. bassolsae, Aedes aegypti and A. albopictus. Essential oils were mainly constituted of β-pinene (>30%) β-caryophyllene (>25%) and germacrene D (>13%). Dorsal-abdomen application of essential oils on triatomines, revealed an efficient LC₅₀ for nymphs of the stages I to III (4 µg/insect), nymphs of the stages IV to V (5–6 µg/insect), and adults (7–8 µg/insect). The LT₅₀ for the stages I to III was between 6 and 8 h, whereas that for the stages IV to V and adults oscillated between 12 and 16 h and 22 to 26 h, respectively. Fumigation experiments performed on nymph V, demonstrated that 300 µg L^?1 air produced 100% mortality after 72 h post-treatment. Among tested volatiles, β-pinene and β-caryophyllene produced a comparable mortality rate (p < 0.01) than that of essential oils in the stages assayed. Essential oils showed strong larvicidal (LC₅₀, 24–36 µg mL^?1) and adulticidal (35–48 µg mL^?1) activities in mosquito species with an LT₅₀ of 4.5 h and 25–35 min, respectively. The evaluation of β-pinene produced a significant mortality rate (p < 0.01) in larvae whereas germacrene D was the most effective volatile (p < 0.01) against adults of both mosquito species. According to our results, β-pinene was the most effective volatile against the four insect species evaluated and its effect was comparable to that of the essential oil.     

The effect of feeding rate on growth of pectoral fin spine of juvenile great sturgeon Huso huso (Linnaeus, 1758)

The effect of two feeding rates (0.5 and 1.5% of total body weight) was assessed on the growth of pectoral fin spines of captive juvenile great sturgeon, Huso huso, after second year of life. The fish received Oxytetracycline (OTC) twice in the first and second years of their lives under basic diet. During the 5‐month experimental period, juveniles (mean 1,187.4 g, 0.1 standard deviation [SD]), n = 50) were reared with two feeding rates under similar conditions in 10 fiberglass tanks (1.5 m³). The fish were fed manually with a commercial diet twice a day (35% Biomar, Nersac, France) throughout the experiment. The OTC marks were distinguished in all pectoral fin spine sections under ultraviolet light. The means of the first and second annular radii were 806.6 µm (27.2 SD) and 2,246.5 µm (50.2 SD), respectively. The marginal increment analysis beyond the second OTC mark revealed a significantly smaller marginal increment for low feeding rate treatment (143.9, 11.2 SD) as compared to the high feeding rate one (269.0, 14.6 SD). The results indicate the slower growth rate in the fish fed the low feeding treatments seen in the pectoral fin spine formation, which can be used as an indicator of recent feeding history in sturgeon juveniles. The best daily feeding rate for great sturgeon of 2,460 g was determined to be 1.5% body weight/day in this study.     

The impact of plant associations on Macrosteles quadrilineatus management in carrots

Habitat diversification can influence the interactions of insects with plants and this can be used in agroecosystems for the management of pest populations. Plant diversification can be achieved through planting crops, such as trap crops, or by adjusting weed management. Aster leafhopper, Macrosteles quadrilineatus Forbes (Hemiptera: Cicadellidae), is a polyphagous species that uses cereals, vegetables, and weeds as host plants. The influence of weeds on M. quadrilineatus abundance was investigated experimentally in carrot [Daucus carota L. cv. Canada (Apiaceae)] field plots by adjusting the level of management of two groups of weeds (broadleaf and grass) and by comparing it to weed‐free plots. The preference of M. quadrilineatus for different cereal and weed species relative to carrots was tested in choice test assays. Habitat context influenced the abundance of M. quadrilineatus in the field experiments. The presence of border crops such as oat, rye, barley, wheat, and triticale did not significantly attract or repel this insect to carrot plots compared to the no‐border treatment. However, spelt‐bordered plots had 42% fewer M. quadrilineatus than three treatments, triticale, wheat, and barley, that had the highest insect abundance. The type of weed management affected M. quadrilineatus abundance in carrot plots, but not the frequency of herbicide application. Plots that had carrot growing with broadleaf‐weeds had about 59% fewer M. quadrilineatus compared with those growing with crabgrass or carrot alone. In the greenhouse choice tests, grasses (e.g., cereals) attracted and broadleaf‐weeds repelled M. quadrilineatus relative to carrots. In summary, carrot growers may be able to manage this pest by reducing the interaction of cereal cover crops with carrots and eliminating grassy weeds in commercial production fields.     

Effect of lead on the physiological,biochemical and ultrastructural properties of Leucaena leucocephala

• Heavy metals are characterised by a relatively high density and cause genotoxic, cytotoxic and mutagenic effects on plants, animals and humans. Lead (Pb) is one of the heavy metals that causes toxicity to plants and animals.
• This experiment was conducted using a hydroponic technique to study the effects of Pb(NO₃)₂ on physiological, biochemical and ultrastructural characteristics in Leucaena leucocephala seedlings. Plants were grown in a growth chamber for 21 days in Hoagland’s solution supplemented with 0 (control), 25, 50, 100, 300, 500 and 700 µm Pb(NO₃)₂.
• Shoot heights as well as root lengths decreased significantly in Pb‐treated plants with 300, 500 and 700 µm . In Pb‐treated plants with high Pb concentrations, photosynthesis rate (P_N), stomatal conductance (g_s) and transpiration rate (E) decreased. Total protein and carbohydrate content in Pb‐treated plants with 300, 500 and 700 µm increased significantly in leaves. Moreover, in Pb‐treated plants with 300, 500 and 700 µm Pb(NO₃)₂, mesophyll cells had enlarged chloroplasts with disrupted thylakoid membranes associated with large starch grains. In contrast, Pb treatments with 25, 50 µm and 100 µm were not toxic to the plants. Thick sections of roots of Pb‐treated plants with 300, 500 and 700 µm Pb showed distinct changes in structure of epidermal and cortical cells. Moreover, thin sections of roots of Pb‐treated plants with 300, 500 and 700 µm Pb had thickened walls of xylem cells.
• These results will shed more light in understanding the effects of heavy metal stress on plants.

     

Effectiveness of aromatic plant species for repelling and preventing oviposition of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae)

This study assesses the effectiveness of essential oils for repelling whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae), and deterring oviposition. We evaluated essential oils from four plants: Curcuma longa (Zingiberaceae), Litsea cubeba (Lauraceae), Piper nigrum (Piperaceae) and Zanthoxylum bungeanum (Rutaceae), under laboratory and glasshouse conditions. In the laboratory, ethanol‐extracted essential oils were applied uniformly on both sides of a tomato leaf in the treatment groups, and the same volume of liquid but without essential oil was used for the control. Results showed that essential oils from L. cubeba, P. nigrum, C. longa and Z. bungeanum provided some repellency of whiteflies and reduced oviposition. Oils from L. cubeba repelled adult females (69.14% and 62.49% repellency at 24 and 48 hr, respectively), and oils from C. longa deterred oviposition (68.46% and 65.94% at 24 and 48 hr exposures, respectively). In the glasshouse, potted tomato plants with eight leaves were sprayed with treatments and control material. Five‐day‐old adult female whiteflies were released in a cage between both pots. Essential oils from L. cubeba repelled females 54.77 at 24 hr and P. nigrum 44.37% at 48 hr whereas oils from P. nigrum reduced oviposition by 43.35 at 24 hr and Z. bungeanum 21.08% at 48 hr, under glasshouse conditions. Essential oils show potential to be used to reduce feeding and oviposition, and thus being important components of integrated pest management programs. However, future research is needed to investigate individual compounds of these oils against whitefly, and to enhance their effectiveness.     

Seed germination and seedling survival traits of invasive and non-invasive congeneric <Emphasis Type="Italic">Ruellia</Emphasis> species (Acanthaceae) in Yucatan,Mexico

We compared requirements for seed germination and seedling establishment for Ruellia nudiflora, an invasive species in Yucatan, Mexico with those of the congeneric non-invasive R. pereducta. Germination and seedling survival rates were higher for R. nudiflora than for R. pereducta under high light. Additionally, the ranges of temperature and water potential that allow germination for R. nudiflora were much broader than those of R. pereducta. Seedlings of R. nudiflora exhibited higher survival to drought by shedding their leaves during drought, an important strategy in environments under extreme drought. Seedlings of R. nudiflora also exhibited higher extreme temperature tolerance than R. pereducta seedlings. Overall, traits exhibited by R. nudiflora such as ability to germinate under a wide range of conditions, adaptation to environmental stress and high tolerance to environmental heterogeneity during the seedling stage, have been repeatedly recognized as determinants of colonization success of invasive species in open disturbed areas.     

Antagonistic,Anti-oxidant,Anti-inflammatory and Anti-diabetic Probiotic Potential of Lactobacillus agilis Isolated From the Rhizosphere of the Medicinal Plants

Potential probiotic bacteria can be used as a biotherapeutic agent and a sustainable alternative to antibiotics, as an anti-oxidative, anti-inflammatory, and anti-diabetic agent without causing any serious side effects. Mostly human-friendly Lactic acid bacteria (LAB) have been isolated from the animal-human origin to be used as biotherapeutic agents or to produce useful metabolites (nutraceutical). However, less information is known about the role of medicinal plants associated LAB as biotherapeutic agents. The isolation of 115 human-friendly Lactobacillus strains was done from the rhizosphere of the medicinal plants Ocimum tenuiflorum, Azadirachta indica, Ficus carica. The obtained bacteria were then tested for their safe status before being using it for a beneficial purpose. Out of 115 strains, 29 (25%) were negative for blood hemolytic activities. Among these 29 isolates, three isolates did not show a breakdown of gelatin and were recognized as safe. Antibiotic resistance assay showed resistance of two of them against antibiotics discs of Streptomycin (10 µg), Ciprofloxacin (20 µg), Vancomycin (30 µg), Metronidazole (10 µg), Ampicillin (5 µg), Chloramphenicol (30 µg), Kanamycin (30 µg), Erythromycin (15 µg), Penicillin (10 µg) and Tetracycline (30 µg). The bacterial isolate (T-2) was found safe that was identified as Lactobacillus agilis by sequence analysis of 16 s rRNA gene and processed in vitro as an anti-bacterial, anti-oxidant, anti-diabetic, and anti-inflammatory agent. Free radical scavenging activities and inhibition of α-amylase activities for Lactobacillus agilis were found relative to standard drug values as 68% and 73% and 51.3% and 65.3%, respectively. The in-vitro anti-inflammatory assay showed 61.6% (Lactobacillus agilis) while showed 69% (aspirin) activity for denaturation albumin protein. The results suggested that Lactobacillus agilis can be used as a potential probiotic strain as well as can be used to produce nutraceuticals.     

Chalcone analogues: Synthesis,activity against Meloidogyne incognita,and in silico interaction with cytochrome P450

To contribute the development of new products to control plant‐parasitic nematodes, 12 chalcone analogues were synthesized and screened for activity against Meloidogyne incognita. Three caused mortality greater than negative controls in second‐stage juvenile M. incognita, with values varying from 19.9% to 100%. The most active chalcone analogue was (1E,4E)‐1,5‐di(4‐nitrophenyl)‐2‐butylpenta‐1,4‐dien‐3‐one (compound 6 ), which had an LC₅₀ value of 41 µg/ml. Under the same conditions, the commercial nematicide Carbofuran^® (2,2‐dimethyl‐2,3‐dihydro‐1‐benzofuran‐7‐yl methylcarbamate) presented an LC₅₀ equal to 101 µg/ml. When this chalcone analogue was applied to tomato plants infested with M. incognita, reductions in the numbers of galls and eggs of 51% and 68% were observed, respectively. According to in silico studies, the enzyme target of compound 6 in M. incongita is cytochrome P450, which is important for the oxidation of several substances in the nematode. Therefore, compound 6 is potentially useful for the development of new products to control M. incongita.     

Antifungal Susceptibility Profile of Candida Albicans Isolated from Vulvovaginal Candidiasis in Xinjiang Province of China

We investigated the antifungal susceptibility profiles of 207 independent Candida albicans strains isolated from patients with vulvovaginal candidiasis (VVC) in Xinjiang Province of China. Using CLSI M27-A3 and M27-S4 guidelines, anidulafungin and micafungin were the most active drugs against C. albicans showing an MIC₅₀/MIC₉₀ corresponding to 0.016/0.0313 µg/mL, followed by caspofungin (0.25/0.25 µg/mL), posaconazole (0.125/0.5 µg/mL), ravuconazole (0.063/1 µg/mL), itraconazole (0.125/1 µg/mL), amphotericine B (0.5/1 µg/mL), isavuconazole (0.063/2 µg/mL), 5-flucytosine (1/2 µg/mL), voriconazole (0.125/4 µg/mL), and fluconazole (0.5/4 µg/mL). 96.1% (199)–100.0% (207) isolates were sensitive to the three echinocandins tested, amphotericine B and 5-flucytosine. The in vitro activity of triazoles against all isolates tested was variable; itraconazole and voriconazole had reduced the activity to almost half of the isolates (55.1% (114) and 51.2% (106) susceptible, respectively). Fluconazole was active against 76.3% (158) isolates tested. The new triazoles ravuconazole, isavuconazole and posaconazole showed good in vitro potency against 89.9% (186)–95.2% (197) of isolates with the geometric mean MIC (µg/mL) of 0.10, 0.12 and 0.14 µg/mL, respectively. In conclusion, our study indicates that for effective management of systemic candidiasis in Xinjiang Province of China, it is important to determine the susceptibility profiles of isolated C. albicans from patients with VVC.

     

In vitro antioxidant and cytotoxic activities of polyherbal extracts from Vetiveria zizanioides,Trichosanthes cucumerina,and Mollugo cerviana on HeLa and MCF-7 cell lines

Various metabolites exist in the medicinal plants have lot of potential to cure various diseases and disorders. Plants such as, Vetiveria zizanioides, Trichosanthes cucumerina, and Mollugo cerviana were collected from Western Ghats, Tamilnadu, India. Phytochemicals were extracted from these plants using various organic solvents and tested against Gram-positive and Gram-negative bacteria. The phytochemicals such as, carbohydrate, alkaloids, steroids, saponins, flavonoids and tannin were detected from these medicinal plants. Among the extracts, methanol showed potent activity and this solvent was used to extract polyherbal medicinal plants. Methanol extract of V. zizanioides was found to be highly active against E. coli (27 ± 2 mm), P. mirabilis (19 ± 3 mm) and B. subtilis (18 ± 2 mm). Ethyl acetate extract showed high activity against E. coli (24 ± 2 mm), P. mirabilis (22 ± 3 mm) and B. subtilis (20 ± 1 mm). These three plants were taken at 1:1:1 ratio and extracted with methanol at 1:10 ratio and synergistic activity was tested against bacterial pathogens. Synergistic activity of polyherbal extract was analyzed. The extracted crude herbal medicine was found to be effective against Staphylococcus aureus, E. coli, Enterbacter sp., Pseudomonas aeruginosa, Bacillus subtilis and Proteus mirabilis. The zone of inhibition was 33 ± 3 mm, 17 ± 2 mm, 22 ± 2 mm, 40 ± 2 mm, 33 ± 1 mm and 38 ± 2 mm zone of inhibition against E. coli, S. aureus, P. aeruginosa, P. mirabilis, B. subtilis and Enterobacter sp. Polyherbal extract was found to be highly effective against P. mirabilis and Enterobacter sp. MIC values of polyherbal extract ranged from 29 ± 2.5 µg/ml to 34 ± 2.5 µg/ml. MIC value was found to be less against P. mirabilis and was high against S. aureus. Antioxidant property varied between 49 ± 3% and 95.3 ± 2%. At 20 µg/ml antioxidant activity was reported as 49 ± 3% and it was increased at higher concentrations of polyherbal extract. Two cell lines (HeLa and MCF cell lines) were selected to analyze cytotoxic activity of polyherbal extract. The methanol extract of polyherbal fraction showed cytotoxicity against these two cell lines. The LC50 value was 467 ± 2.9 µg/ml against HeLa cell line and >800 µg/ml against MCF-7 cell lines. The polyherbal extract showed antibacterial, antioxidant and anticancer activities.     

Effect of mechanical damage on emission of volatile organic compounds from plant leaves and implications for evaluation of host plant specificity of prospective biological control agents of weeds

Assessment of host plant specificity is a critical step in the evaluation of classical biological control agents of weeds which is necessary for avoiding possible damage to non-target plants. Volatile organic compounds (VOCs) emitted by plants likely play an important role in determining which plants attract and are accepted by a prospective arthropod agent. However, current methods to evaluate host plant specificity usually rely on empirical choice and no-choice behavioural experiments, with little knowledge about what chemical or physical attributes are stimulating the insect. We conducted experiments to measure the quantitative and qualitative effects on emission of VOCs caused by simple mechanical damage to leaves of plants known to differ in suitability and attractiveness to a prospective agent. More VOCs were detected from damaged than from undamaged leaves for all three species tested. Discriminant analysis was able to correctly distinguish the taxonomic identity of all plants based on their VOC profiles; however, the VOCs that discriminated species among undamaged leaves were completely different from those that discriminated among damaged leaves. Thus, damaged and undamaged plants present different VOC profiles to insects, which should be considered when conducting host plant specificity experiments. An unacceptable non-target plant, Centaurea cineraria, emitted all except one of the VOCs that were emitted by its preferred host plant, Centaurea solstitialis, indicating the importance of compounds that are repellant in host plant specificity. Centaurea cyanus emitted fewer VOCs than C. solstitialis, which suggests that it lacked some VOCs important for host plant recognition.     

Somatic embryogenesis and shoot organogenesis from the hypocotyl slices and free radical scavenging activity of regenerants of collard greens (Brassica oleracea L. var. acephala)

Brassica oleracea var. acephala is an important leafy vegetable that has been widely consumed as a high-nutrient, low-calorie food. Because of the plant’s biennial and self-incompatibility nature, biotechnological approaches are alternative way for propagation and breeding improvements. Since tissue culture studies have been focused in other B. oleracea representatives, the aim of the present study was to achieve effective regeneration protocol distinctive for collard greens, and evaluate the total phenolic content and antioxidant activity of regenerants. The effect of 3 cytokinins [thidiazuron (TDZ), 6-benzyladenine (BA) and 6-furfuryladenine (kinetin, KIN)] at increasing concentrations (0, 5, 7.5, 10, 20 or 30 µM) in combination with tenfold lower concentration of 1-naphtaleneacetic acid (NAA) (0, 0.5, 0.75, 1, 2 or 3 µM, respectively) on the regeneration from hypocotyl slices was studied. Histological analysis revealed the two regeneration pathways, somatic embryogenesis and shoot organogenesis, simultaneously occurred in the same explant, regardless of the cytokinin/NAA combinations used. The regeneration frequency of 95.9%, with 7.5 morphogenic structures regenerated per explant, and the healthy appearance of regenerated plants indicated the optimal combination 20 µM TDZ?+?2 µM NAA. TDZ at 5 µM provided the high somatic embryo proliferation rate by generation of secondary embryos (7.79) along with the lowest rate of their abnormalities. Embryo-developed plants were successfully acclimatised (above 90%). The plants regenerated and proliferated on TDZ-containing media had higher total phenolic content that correlated with the highest free radical scavenging activity (IC₅₀?=?19.09 µg ml^??1).

     

Detection and molecular characterization of carbendazim-resistant Colletotrichum truncatum Isolates causing anthracnose of soybean in Thailand

A loss of fungicide efficacy, particularly for carbendazim, was noted in soybean fields in Thailand and was considered to be due to the development of Colletotrichum truncatum resistance. The carbendazim sensitivity of C. truncatum populations isolated from various soybean fields in Thailand was thus evaluated with in vitro sensitivity assays and molecular characterization of mutations in the sequences of the ß₂-tubulin (TUB2) gene that confer carbendazim resistance in the pathogen. Among 52 isolates, 46 isolates were classified as highly resistant (HR) to carbendazim (EC₅₀ > 1,000 µg/ml). All HR isolates grew on PDA amended with carbendazim at 1,000 µg/ml. Six isolates were classified as carbendazim sensitive (S) (EC₅₀ < 1 µg/ml). Mycelial growth on PDA amended with 1 µg/ml carbendazim was inhibited by over 50% compared with growth on PDA alone. When a partial TUB2 gene from the isolates was amplified and analysed using predicted amino acid sequences, an alteration from glutamic acid to alanine at codon 198 (E198A) was found in 45 HR isolates for which the EC₅₀ was higher than 2000 µg/ml. This mutation resulted from a nucleotide substitution from adenine to cytosine (GA G → GC G). The other HR isolate, CtPhS_1, with EC₅₀ of 1,127 µg/ml, had an alteration at codon 200 (F200Y) (TT C → TA C).     

Compared with conventional PCR assay,qPCR assay greatly improves the detection efficiency of predation

Studies of predation can contribute greatly to understanding predator–prey relationships and can also provide integral knowledge concerning food webs and multi‐trophic level interactions. Both conventional polymerase chain reaction (cPCR) and quantitative PCR (qPCR) have been employed to detect predation in the field because of their sensitivity and reproducibility. However, to date, few studies have been used to comprehensively demonstrate which method is more sensitive and reproducible in studies of predation. We used a Drosophila melanogaster‐specific DNA fragment (99 bp) to construct a tenfold gradient dilution of standards. Additionally, we obtained DNA samples from Pardosa pseudoannulata individuals that fed on D. melanogaster at various time since feeding. Finally, we compared the sensitivity and reproducibility between cPCR and qPCR assays for detecting DNA samples from feeding trials and standards. The results showed that the cPCR and qPCR assays could detect as few as 1.62 × 10³ and 1.62 × 10¹ copies of the target DNA fragment, respectively. The cPCR assay could detect as few as 48 hr post‐feeding of the target DNA fragment. But the qPCR assay showed that all spiders were positive after consuming prey at various time intervals (0, 24, 48, 72, and 96 hr). A smaller proportion of the technical replicates were positive using cPCR, and some bands on the agarose gel were absent or gray, while some were white and bright for the same DNA samples after amplification by cPCR. By contrast, a larger proportion of the technical replicates were positive using qPCR and the coefficients of variation of the Ct value for the three technical replicates of each DNA sample were less than 5%. These data showed that qPCR was more sensitive and highly reproducible in detecting such degraded DNA from predator's gut. The present study provides an example of the use of cPCR and qPCR to detect the target DNA fragment of prey remains in predator's gut.