Fungicide resistance of Botrytis cinerea in tomato greenhouses in the Canary Islands and effectiveness of non-chemical treatments against gray mold

Tomato greenhouses in the Canary Islands, Spain, were surveyed to estimate frequencies of resistance to benzimidazoles, dicarboximides, anilinopyrimidines and N-phenylcarbamates in Botrytis cinerea. Resistance to carbendazim, iprodione, pyrimethanil and diethofencarb was found in 74.2, 86.4, 28.8 and 31.8 % of isolates, respectively. Benzimidazole- and anilinopyrimide-resistant isolates were highly resistant, showing EC₅₀ values above 500 µg/ml carbendazim and a mean EC₅₀ value of 28.42 µg/ml pyrimethanil, respectively. By contrast, a low level of resistance was observed among dicarboximide-resistant isolates (mean EC₅₀ value of 1.81 µg/ml iprodione). Phenotypes with double resistance to carbendazim and iprodione, and triple resistance to carbendazim, iprodione and pyrimethanil were the most common, occurring in 36.4 and 28.8 % of isolates. The surveyed greenhouses had never been treated with fenhexamid and Signum? (pre-packed mixture of boscalid and pyraclostrobin), and baseline sensitivities of B. cinerea isolates to these fungicides were determined. The EC₅₀ values were within the range of 0.009–0.795 µg/ml fenhexamid and of 0.014–0.48 µg/ml Signum. In addition, available formulations based on elicitors of plant defense response and biocontrol agents were evaluated against B. cinerea in tomato plants under semi-controlled greenhouse conditions, the yeast Candida sake CPA-1 being able to reduce gray mold significantly when it was applied on petiole wounds and the plants were inoculated 24 h later. Likewise, C. sake was effective against B. cinerea in harvested tomato fruits, yeast-treated tomatoes showed a 70.66 and 30.31 % reduction in the diameters of decay lesions compared with controls after 10 days of storage at 20 and 9 °C, respectively.     

Baseline sensitivities to azoxystrobin and tebuconazole in Sclerotinia sclerotiorum isolates from sunflower in Iran related to sensitivities to carbendazim and iprodione

In this study, sensitivities of 156 Sclerotinia sclerotiorum isolates collected from sunflower fields of West Azarbaijan province, Iran, were assessed to carbendazim and iprodione, and the baseline sensitivities were established for azoxystrobin and tebuconazole. Resistance to carbendazim and iprodione was observed in 53.85% and 4.49% of the isolates, respectively. The 50% effective concentration (EC₅₀) values of azoxystrobin for the isolates ranged from 0.017 to 3.515 μg/ml with a mean of 0.330 μg/ml, and 8.97% of the strains showed low levels of resistance to the fungicide. However, in the presence of salicylhydroxamic acid, all isolates were sensitive to azoxystrobin and EC₅₀ values ranged from 0.015 to 0.263 μg/ml with a mean of 0.086 μg/ml. All isolates were found to be sensitive to tebuconazole, and EC₅₀ values ranged from 0.003 to 0.177 μg/ml with a mean of 0.036 μg/ml. Among the multiple-resistant isolates, the strains exhibiting resistance to both carbendazim and iprodione were detected in the highest frequency (4.49%). No correlation was observed between mycelial growth and aggressiveness with fungicide sensitivity of the isolates suggesting the absence of fitness cost associated with resistance to the studied fungicides. The results indicated that iprodione, azoxystrobin and tebuconazole could be effectively used in rotation or mixture in spray programmes to manage S. sclerotiorum in the region. The baselines established for azoxystrobin and tebuconazole would be useful in monitoring the fungal populations in the province to assess possible shifts in fungicide sensitivity of S. sclerotiorum isolates in the future.     

Mutations of the β‐tubulin gene fragments from carbendazim‐resistant isolates of Pestalotiopsis sp. causing strawberry leaf blight in Chiang Mai,Thailand

Screening of field isolates of Pestalotiopsis sp. from strawberry leaf blight in Thailand identified 56 carbendazim‐resistant isolates. Of 56 isolates, 39 highly resistant (HR) phenotypes grew well on PDA amended with carbendazim even at ≥500 mg/L. Isolates with carbendazim‐resistant phenotype had a conspicuous mutation at particular sites in the beta‐tubulin (β‐tubulin) gene sequence. A β‐tubulin encoding gene from this pathogen was cloned and sequenced. Analysis of the β‐tubulin gene in highly resistant (HR) isolates showed a substitution at codon 79 and 102 from serine (AGT) to lysine (AAA) and valine (GTA) to alanine (GCA), respectively. The detection of such point mutations in the β‐tubulin gene allows the rapid screening to detect carbendazim‐resistant isolates in the field.     

Baseline sensitivity and control efficacy of fluazinam against Sclerotinia sclerotiorum in Henan Province,China

Sclerotinia stem rot, caused by Sclerotinia sclerotiorum, is a devastating disease in Henan Province, of the main rapeseed production areas in China. Fluazinam belongs to the broad‐spectrum phenylpyridinamine fungicides, which have high activity in inhibiting the mycelial growth of S. sclerotiorum. In this study, 191 field isolates were obtained from different oilseed rape fields in Henan Province, before being exposed to fluazinam in 2015. The baseline sensitivity of S. sclerotiorum to fluazinam was established. The effective concentration for 50% inhibition of mycelial growth (EC₅₀) ranged from 0.0019 to 0.0337 μg/ml, and the mean EC₅₀ value was 0.0084 ± 0.0055 μg/ml. The range of the frequency distribution was narrow. The results of a cross‐resistance assay revealed no cross‐resistance between fluazinam and carbendazim, dimethachlone, boscalid or fludioxonil. Field efficacy tests showed that the control efficacies of fluazinam (50% WG) applied at 150, 225 and 300 g ai ha^?1 were 67%, 73% and 88%, respectively. In contrast, the control efficacies of boscalid (50% WG) and carbendazim (50% WP) applied at 225 and 1,500 g ai ha^?1 were 71% and 52%, respectively.     

Baseline sensitivity of Botrytis cinerea to fluazinam and cross‐resistance

Grey mould, caused by the fungus Botrytis cinerea Pers ex Fr., is a very destructive and important disease worldwide. Fluazinam is a phenylpyridinamine fungicide with broad‐spectrum activities. The baseline sensitivity of B. cinerea to fluazinam is yet to be established in Henan Province, China. In this study, a total of 117 field isolates of B. cinerea were collected from 49 commercial greenhouses in different locations of Henan Province, in 2016, and the sensitivities of these isolates to fluazinam were determined based on mycelial growth. The effective concentration for 50% (EC₅₀) values ranged from 0.0038 to 0.0441 μg/ml, and the mean EC₅₀ value was 0.0201 ± 0.0081 μg/ml for mycelial growth. The frequency distribution range presented a unimodal curve. To define the cross‐resistance relationships, the linear correlation coefficients of the EC₅₀ values between fluazinam and carbendazim, procymidone, pyrimethanil or boscalid were analysed. The results showed that no correlation was observed between fluazinam and the other tested fungicides. These results provide important information to growers for the prevention and control of grey mould.     

Baseline sensitivity of Fusarium graminearum from wheat fields in Henan,China, to metconazole and analysis of cross resistance with carbendazim and phenamacril

The filamentous plant pathogenic fungus Fusarium graminearum is one of the most important pathogens causing Fusarium head blight (FHB) in wheat in the Henan Province of China. Metconazole is among the demethylation inhibitor (DMI) fungicides with a higher inhibitory activity on the mycelial growth of F. graminearum. In 2016 and 2017, 119 single spore isolates of F. graminearum, prior to being exposed to metconazole, were recovered from 52 wheat fields near 11 cities in Henan Province. The inhibitory activity of metconazole on the mycelia of the Henan F. graminearum population was determined, and EC₅₀ values were calculated. The range of EC₅₀ values of the Henan F. graminearum population to metconazole was 0.0103 to 0.0775 μg/ml with an average EC₅₀ value of 0.0293 ± 0.0114 μg/ml. The sensitivity frequency distribution curve presented a single peak in a narrow range. No cross-resistance was found between the DMI fungicide metconazole and the benzimidazole fungicide carbendazim or the cyanoacrylate fungicide phenamacril. Therefore, these sensitivity data could be used as the baseline of F. graminearum susceptibility to metconazole in the Henan Province and provide the basis for monitoring metconazole resistance in this area.     

Occurrence of fenhexamid resistance in Botrytis cinerea from greenhouse strawberries in China

This study assessed the fenhexamid sensitivity of 143 Botrytis cinerea isolates collected from greenhouse strawberries in five regions of China between 2012 and 2013, and identified four isolates with moderate levels of resistance: two from the Xinjiang Uygur Autonomous Region and two from Hebei Province. The baseline fenhexamid sensitivity of B. cinerea exhibited a unimodal distribution with a mean EC₅₀ value of 0.20 ± 0.10 μg/ml (SD). The EC₅₀ values of the fenhexamid‐resistant isolates ranged from 0.05 to 0.40 μg/ml. Molecular analysis of the fenhexamid target gene erg27 revealed that the resistant isolates collected from Xinjiang (163‐6 and 163‐22) contained three mutations that led to amino acid changes (V365A, E368D and A378T) known to be associated with fenhexamid resistance, but that the isolates from Hebei lacked any mutations, indicating that an alternative mechanism could be responsible for their resistance. Most of the biological characteristics of the fenhexamid‐resistant isolates, such as mycelial growth, sclerotia production and pathogenicity, did not significantly differ from those of the sensitive ones (p ≤ .05), but it was noted that some of the resistant isolates exhibited reduced rates of sporulation and spore germination. In addition, the resistant isolates exhibited lower osmotic sensitivity than the sensitive ones. The study found no evidence of cross‐resistance with other fungicides, but that there was negative cross‐resistance with procymidone, iprodione, carbendazim and pyraclostrobin, which indicates that the inclusion of these fungicides within an integrated pest management (IPM) programme could help to minimize the risk of fenhexamid resistance developing in B. cinerea.     

Sensitivity determination and resistance risk assessment of Rhizoctonia solani to SDHI fungicide thifluzamide

Laboratory experiments were conducted to determine (a) the baseline sensitivity of Rhizoctonia solani to thifluzamide and (b) the risk of the fungus developing resistance to the fungicide. Thifluzamide sensitivity was assessed for 227 isolates of R. solani collected from 12 provinces of China from 2007 to 2011. One insensitive isolate GD‐1 was obtained from the field, and the EC₅₀ values of the 226 sensitive isolates had a unimodal frequency distribution with a mean of 0.0351 µg mL^?1. Nine resistant mutants were generated using thifluzamide‐amended media or UV radiation in the laboratory. The resistance was stable for all mutants after 10 transfers on PDA medium. Fitness of the most resistant mutants was lower than that of the sensitive isolates, implying a lower competitiveness of the mutants relative to sensitive isolates in field. Cross‐resistance was detected between thifluzamide and the Succinate dehydrogenase inhibitors (SDHIs) fenfuram, carboxin, penflufen and boscalid, but not between thifluzamide and difenoconazole, carbendazim, propiconazol, SYP‐2815 (quinone outside inhibitor (QoI) fungicide developed in China), fluazinam, jinggangmycin, pyrimorph or mepronil. The SDHI fungicide fluopyram did not inhibit R. solani. Taken together, these results suggest that the risk of R. solani developing resistance to thifluzamide is low to moderate.     

In-vitro antioxidant,antimutagenic and cancer cell growth inhibition activities of Rhododendron arboreum leaves and flowers

In the current investigation, the active principles of the methanol extracts of Rhododendron arboreum leaves (MEL) and flowers (MEF) were investigated with the help of ultra-high performance liquid chromatography (UHPLC), amino acid analyzer and gas chromatography mass spectrometry (GC-MS). UHPLC revealed different polyphenols present in the extracts. GC-MS identified 20 phytochemicals in leaves and 17 in the flowers, whereas, amino acid analyzer confirmed 11 amino acids in leaves and 10 in the flowers. The extracts were subjected to the investigation of biological activity through analysis of antioxidant activity in different in vitro assays, antimutagenic activity in Ames assay and cancer cell growth inhibition activity by MTT (3-4,5 dimethylthiazol-2,5 diphenyltetrazolium bromide) assay. MEL showed higher antioxidant activity in lipid peroxidation inhibition assay (95.32 ± 0.37%) than MEF (77.09 ± 4.17%) with IC₅₀103.6 µg/ml for MEL and 271.17 µg/ml for MEF. In nitric oxide scavenging assay, an activity of 94.46 ± 0.32% (IC₅₀ 150.13) was observed in MEF followed by 83.71 ± 0.74% (IC₅₀ 179.52) in MEL. The antimutagenic activity of both the extracts was evaluated against sodium azide, 4-nitro-O-phenylenediamine and 2-aminofluorene mutagens in TA-98 and TA-100 strains of Salmonella typhimurium. The analysis was carried out using pre- and co-incubation modes. However, both extracts were observed to possess considerable antimutagenic activity against different known mutagens, flowers came out to be more effective than the leaves in terms of % inhibition. The extracts also exhibited significant cancer cell growth inhibition activity, when tested against 3 cancer cell lines namely, Human cervical cancer cell line (HeLa), Breast cancer cell line (MCF7) and Lung cancer cell line (A549). In case of HeLa and A549, MEL showed higher activity of 64.62 ± 2.65 and 75.08 ± 1.68% as compared to 53.11 ± 2.84 and 45.92 ± 2.43% in MEL, respectively. The EC₅₀ values for MEL in HeLa and A549 were noted to be 232.76 and 155.38 µg/ml, respectively, whereas, MEF had EC₅₀ of 395.50 µg/ml in HeLa and 660.26 µg/ml in A549. Further, MEF showed higher cytotoxicity in MCF7 cell line (84.93 ± 1.17%) followed by the MEL (73.57 ± 1.27%) with EC₅₀ value of 95.16 µg/ml for MEF followed by 172.19 µg/ml for MEL. The biological activities of the extracts can be attributed to the phyto-constituents identified by sophisticated instruments.The biological activities of the extracts can be attributed to the active principles identified by sophisticated instruments.     

Population Structure and Development of Resistance to Hymexazol Among Fusarium solani Populations from Date Palm,Citrus and Cucumber

A study was conducted to investigate genetic diversity and sensitivity to hymexazol among 80 isolates of Fusarium solani complex obtained from date palm (30), citrus (31) and cucumber (19). Characterization based on sequences of the EF1α and ITS rRNA showed that isolates belong to F. solani complex MLST type 3 + 4. AFLP analysis produced 980 polymorphic loci, 80 AFLP genotypes and moderate levels of genetic diversity (H = 0.2494). Clustering of the isolates was not related to the host or the geographical origin of the isolates. Analysis of molecular variance (amova ) indicated the existence of a low level of genetic differentiation among populations obtained from different hosts (F_st = 0.0162) and regions (F_st = 0.0066). This may provide evidence for frequent movement of inoculum among hosts and regions in Oman, which could be attributed to cultural practices employed by farmers. Isolates of F. solani displayed variation in sensitivity to hymexazol, with EC₅₀ values ranging from 2 to 5745 μg/ml (mean = 878 μg/ml); 19% of the isolates have an EC₅₀ value of more than 1000 μg/ml. Findings are discussed in terms of the factors that affect diversity in F. solani isolates. The study reports for the first time the development of resistance to hymexazol among F. solani isolates from date palm, citrus and cucumber.     

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.

     

Effect of azoxystrobin and kresoxim‐methyl on rice blast and rice grain yield in China

Sensitivity to azoxystrobin and kresoxim‐methyl of 80 single‐spore isolates of Magnaporthe oryzae was determined. The EC₅₀ values for azoxystrobin and kresoxim‐methyl in inhibiting mycelial growth of the 80 M. oryzae isolates were 0.006–0.056 and 0.024–0.287 µg mL^?1, respectively. The EC₅₀ values for azoxystrobin and kresoxim‐methyl in inhibiting conidial germination of the M. oryzae populations were 0.004–0.051 and 0.012–0.105 µg mL^?1, respectively. There was significant difference in sensitivity to azoxystrobin or kresoxim‐methyl between the tested isolates representing differential sensitivity to carbendazim (MBC) and kitazin P (IBP); however, there was no correlation between this difference in sensitivity to azoxystrobin or kresoxim‐methyl and sensitivity to MBC or IBP, indicating that there was no cross‐resistance between azoxystrobin or kresoxim‐methyl and MBC or IBP. In the protective and curative experiments, kresoxim‐methyl exhibited higher protective and curative activity than azoxystrobin when applied at 150 and 250 µg mL^?1 accordingly, while azoxystrobin exhibited stronger inhibitory activity against M. oryzae isolates than that of kresoxim‐methyl in the in vitro test. The results of field experiments also suggested that both azoxystrobin and kresoxim‐methyl at 187.5 g.a.i. ha^?1 gave over 73% control efficacy in both sites, exhibiting excellent activity against rice blast. Taken together, azoxystrobin and kresoxim‐methyl could be a good substitute for MBC or IBP for controlling rice blast in China, but should be carefully used as they were both at‐risk.     

In vitro sensitivity of Verticillium fungicola to selected fungicides

Twenty isolates of Verticillium fungicola var. fungicola collected from diseased fruit-bodies of Agaricus bisporus from prochloraz-treated crops, were exposed to a range of concentrations of six chemicals (benomyl, chlorothalonil, formaldehyde, iprodione, prochloraz-Mn-complex and prochloraz + carbendazim) in vitro. EC₅₀ values were determined for each fungus-fungicide combination. All isolates were more sensitive to prochloraz-Mn-complex (EC₅₀ values less than 5 mg 1^–1) than to the remainder fungicides, and only seven isolates were moderately sensitive (EC₅₀ values between 5 and 50 mg 1^–1) to prochloraz + carbendazim. All isolates were moderately sensitive to formaldehyde, whereas the majority of isolates were very resistant to the other three fungicides (benomyl, chlorothalonil and iprodione).     

Novel anthranilic amide derivatives bearing the chiral thioether and trifluoromethylpyridine: Synthesis and bioactivity

Ten anthranilic amides bearing skeletons of chiral thioether and trifluoromethylpyridine (5a-5j) were designed and synthesized. Bioassays indicated that some of compounds had excellent insecticidal activity. For example, compounds 5a, 5e and 5g had the median lethal concentrations (LC₅₀) against Plutella xylostella of 7.3, 8.7 and 8.1 µg/mL respectively. The LC₅₀ of 5a against Ostrinia nubilalis and Pseudaletia separata were 21.7 and 44.2 µg/mL respectively. Anti-TMV tests indicated that some compounds also showed good antiviral activity. For instance, the curative activities of compounds 5b and 5e were 57.2% and 63.6%, and with half maximal effective concentration (EC₅₀) of 304.5 and 203.0 µg/mL, respectively, which were much higher than these of ribavirin (39.4%, EC₅₀ = 819.8 µg/mL) and ningnanmycin (56.2%, EC₅₀ = 361.4 µg/mL). The molecular docking between the most active compounds and ryanodine receptor of the Plutella xylostella were also discussed. Those results indicated that the novel anthranilic amide derivatives in present work were worthy of further research and development as novel pesticides.     

Establishment of an EC50 database of pesticides using a Vibrio fischeri bioluminescence method

An EC₅₀ database was established to assess the acute toxicity of 16 PESTANAL pesticide standards and of seven pesticide commercial formulations using a Vibrio fischeri bioluminescence method. Half maximal effective concentration ( EC₅₀) is defined as the concentration of pollutant (in this case, pesticide) destroying 50% of the bacteria population and causing 50% bioluminescence inhibition, after a specified exposure time. Linear curves of bioluminescence inhibition versus pesticide concentration and EC₅₀ values were obtained for exposure times (t) of 5 or 15 min for these pesticides. The EC₅₀ values ranged from 6.90 × 10^?4 to 0.83 mg/ml (t = 5 min), and from 9.00 × 10^?4 to 0.37 mg/ml (t = 15 min) for pesticide standards, plus from 0.0077 to 0.74 mg/ml (t = 5 min), and from 0.0076 and 0.57 mg/ml (t = 15 min) for pesticide commercial formulations. The EC₅₀ database allowed classification of the pesticides under study into three categories according to their toxicity: very toxic, toxic and moderately toxic. These results demonstrated that the establishment of an EC₅₀ database and of linear curves of bioluminescence inhibition versus the pesticide concentration resulted in very important and irreplaceable tools to estimate the global and individual toxicity of pesticides present in environmental samples.     

Characterization of adaptability components of Brazilian isolates of Macrophomina pseudophaseolina

Macrophomina pseudophaseolina is a new Macrophomina species reported on different crop and weed species in Brazil, India and Senegal, but to date there are no studies about its adaptability components. In this work, a collection of 62 M. pseudophaseolina isolates obtained from roots of the weed species Trianthema portulacastrum and Boerhavia diffusa collected in Northeastern Brazil, was used to: (a) study the effect of temperature and salinity on mycelial growth, (b) to determine their sensitivity to the fungicide carbendazim and (c) to assess their aggressiveness on melon and watermelon seedlings. Results showed variability among M. pseudophaseolina isolates. The optimum temperature for mycelial growth ranged between 26.4 and 38.1ºC. NaCl reduced the in vitro growth of all isolates, which were also highly sensitive to the fungicide carbendazim, exhibiting EC₅₀ values ranging from 0.013 to 0.089 mg/L a.i. Disease severity values on melon and watermelon seedlings showed that M. pseudophaseolina isolates were more aggressive in melon than in watermelon. Information about adaptability components of M. pseudophaseolina obtained in this study could be incorporated on breeding programs for melon and watermelon crops.     

Sensitivity to boscalid in field isolates of Sclerotinia sclerotiorum from rapeseed in Henan Province,China

Sclerotinia stem rot caused by Sclerotinia sclerotiorum is an important disease of oilseed rape in Henan province of China. Boscalid belongs to succinate dehydrogenase inhibitor (SDHI) fungicides, many of which have strong antifungal activity against S. sclerotiorum. In 2015, a total of 175 isolates of S. sclerotiorum were collected from diseased oilseed rape plants in seven different regions of Henan Province. The EC₅₀ values of 175 isolates of S. sclerotiorum to boscalid ranged from 0.0073 to 0.3880 μg ml^?1, and the mean EC₅₀ value was 0.15 ± 0.09 μg ml^?1. The frequency distribution was unimodal. There was no cross‐resistance between boscalid and carbendazim, procymidone, iprodione, dimethachlone, fludioxonil or fluazinam. Field experiments showed that control efficacies of treatments with boscalid (50% WG) at 225, 300 and 375 g ai ha^?1 were 71%, 81% and 90%, respectively. In contrast, the control efficacy of carbendazim (50% WP) at 1,500 g ai ha^?1 was only 52%.     

Anti‐HSV‐1 and HSV‐2 Flavonoids and a New Kaempferol Triglycoside from the Medicinal Plant Kalanchoe daigremontiana

Kalanchoe daigremontiana (Crassulaceae) is a medicinal plant native to Madagascar. The aim of this study was to investigate the flavonoid content of an aqueous leaf extract from K. daigremontiana (Kd), and assess its antiherpetic potential. The major flavonoid, kaempferol 3‐O‐β‐d ‐xylopyranosyl‐(1 → 2)‐α‐l ‐rhamnopyranoside ( 1 ), was isolated from the AcOEt fraction (Kd‐AC). The BuOH‐soluble fraction afforded quercetin 3‐O‐β‐d ‐xylopyranosyl‐(1 → 2)‐α‐l ‐rhamnopyranoside ( 2 ) and the new kaempferol 3‐O‐β‐d ‐xylopyranosyl‐(1 → 2)‐α‐l ‐rhamnopyranoside‐7‐O‐β‐d ‐glucopyranoside ( 3 ), named daigremontrioside. The crude extract, Kd‐AC fraction, flavonoids 1 and 2 were evaluated using acyclovir‐sensitive strains of HSV‐1 and HSV‐2. Kd‐AC was highly active against HSV‐1 (EC₅₀ = 0.97 μg/ml, SI > 206.1) and HSV‐2 (EC₅₀ = 0.72 μg/ml, SI > 277.7). Flavonoids 1 and 2 showed anti‐HSV‐1 (EC₅₀ = 7.4 μg/ml; SI > 27 and EC₅₀ = 5.8 μg/ml; SI > 8.6, respectively) and anti‐HSV‐2 (EC₅₀ = 9.0 μg/ml; SI > 22.2 and EC₅₀ = 36.2 μg/ml; SI > 5.5, respectively) activities, suggesting the contribution of additional substances to the antiviral activity.     

Sprouted and Freeze‐Dried Wheat and Oat Seeds – Phytochemical Profile and in Vitro Biological Activities

This research was carried out to study phytochemical profile, in vitro antioxidant capacity, reducing power, anti‐hyperglycemic, anti‐inflammatory activities and simulated gastrointestinal digestion of 7‐day old cereal sprouts: spelt wheat ‘Nirvana’ (WSSpe), wheat ‘Simonida’ (WSSim), oat ‘Golozrni’ (OSG) and oat ‘Jadar’ (OSJ). OSG expressed significantly higher (P ≤ 0.05) total phenols (TPC) and flavonoids content (TFC), antioxidant capacities (DPPH and ABTS assays) and reducing power (EC₅₀^DPPH = 2.12 mg/ml; EC₅₀^ABTS = 0.87 mg/ml; EC_0.5^RP = 12.24 mg/ml) as well as anti‐hyperglycemic activity (EC₅₀^AHgA = 0.96 mg/ml). WSSpe had the highest content of chlorophyll (131.23 mg/100 g) and carotenoids (22.84 mg/100 g). WSSim possessed the most potent anti‐inflammatory activity (2.71 mg/ml), though not significantly different from OSG (2.77 mg/ml). The in vitro simulation of gastro‐intestinal digestion showed higher release of phenolic compounds in intestinal than in gastric fluid.     

Utility of internally transcribed spacer region of rDNA (ITS) and β‐tubulin gene sequences to infer genetic diversity and migration patterns of Colletotrichum truncatum infecting Capsicum spp.

Anthracnose is among the most economically important diseases affecting pepper (Capsicum spp.) production in the tropics and subtropics. Of the three species of Colletotrichum implicated as causal agents of pepper anthracnose, C. truncatum is considered to be the most destructive in agro‐ecosystems worldwide. However, the genetic variation and the migration potential of C. truncatum infecting pepper are not known. Five populations were selected for study and a two‐locus (internally transcribed spacer region, ITS1‐5.8S‐ITS2, and β‐tubulin, β‐TUB) sequence data set was generated and used in the analyses. Sequences of the ITS region were less informative than β ‐ tubulin gene sequences based on comparisons of DNA polymorphism indices. Trinidad had the highest genetic diversity and also had the largest effective population size in pairwise comparisons with the other populations. The Trinidad population also demonstrated significant genetic differentiation from the other populations. AMOVA and STRUCTURE analyses both suggested significant genetic variation within populations more so than among populations. A consensus Maximum Likelihood tree based on β‐TUB gene sequences revealed very little intraspecific diversity for all isolates except for Trinidad. Two clades consisting solely of Trinidad isolates may have diverged earlier than the other isolates. There was also evidence of directional migration among the five populations. These findings may have a direct impact on the development of integrated disease management strategies to control C. truncatum infection in pepper.