A novel rhizobacterium Bk7 for biological control of brown sheath rot of rice caused by Pseudomonas fuscovaginae and its mode of action

The study presents the first report on biocontrol of brown sheath rot disease of rice caused by Pseudomonas fuscovaginae using rhizo-bacterial isolate Bacillus amyloliquefaciens Bk7. Four potential bioactive antagonists were selected from 120 Bacillus isolates. Results obtained from in vitro laboratory assay showed that rhizosphere bacterial strain Bk7 and its metabolites significantly suppressed the growth of Pseudomonas fuscovaginae with 93 % efficacy. In glasshouse experiments, strain Bk7 exhibited biocontrol efficacy of 76.6 % by reducing the disease incidence to 16.9 %, compared to 72.8 % observed in control treatment. In addition, the isolate Bk7 showed the growth promotion efficacy of plant height (GPE, 46.4 %) and fresh weight (GPE, 84.3 %). Characterization of isolate Bk7 revealed its strong capability for biofilm formation, inorganic phosphate solubilization and production of high amounts of Indole-3 acetic acid, siderophores and ammonia in vitro. Results obtained from multiplex PCR assay confirmed the presence of five lipopeptide biosynthetic gene markers (srfAA, fenD, bmyB, bacA and ituC) in the genome of strain Bk7. Moreover, Real-time qPCR of these genes demonstrated that surfactin, iturin and bacylisin coding genes were highly expressed in response to P. fuscovaginae exposure in vitro. Rhizosphere bacterial strain Bk7 was identified as B. amyloliquefaciens strain Bk7 based on the analysis of 16S rDNA internal transcribed spacer sequences and a fatty acid methyl ester analysis. The results obtained from this study showed the potential usefulness of Bk7 as a biocontrol agent in disease control of rice brown sheath rot.     

Quantifying cropping practices in relation to inoculum levels of Fusarium graminearum on crop stubble

This study was conducted in 58 producer‐field locations in Manitoba from 2003 to 2006 to understand how cropping practices influence Fusarium graminearum inoculum levels on stubble of various crops, including wheat, collected from the soil surface. Colonies per m² (CN) were determined and converted to base‐10 logarithm values (log₁₀CN). Mean log₁₀CN of the sampled field for various crops and groups of crops grown in the 3 years prior to sampling were tested to find significant differences. Average log₁₀CN values were also used to determine significant differences between tillage systems and the effect of number of years. Average log₁₀CN values for zero and minimum tillage systems were not different but were significantly higher than values for conventional tillage. A series of three crop rotation scenarios were tested using weighted log₁₀CN values for crop, tillage, their interaction and their squared terms in step‐wise regression models to identify which model was the best predictor of log₁₀CN. This was selected as the cropping practice index (CPI) model and was expressed as: CPI = 1·98423 + 0·55975 (C₂ × C₁ × T)² + 0·4390 (C₂ × T)², where C₁, C₂ and T represent the weighted log₁₀CN values for crops grown 1 and 2 years previously and tillage system, respectively. R² value for this model was 0·933 (P < 0·0001). The reliability of the CPI model was tested using jack‐knife full cross‐validation regression. The resulting R² was 0·899. The CPI model was tested using data collected from seven wheat fields in 2006 (R² = 0·567). The relationship between CPI and FHB index (R² = 0·715) was significant.     

Relationship between disease severity and escape of Pseudoperonospora cubensis sporangia from a cucumber canopy during downy mildew epidemics

Fundamental to the development of models to predict the spread of cucurbit downy mildew is the ability to determine the escape of Pseudoperonospora cubensis sporangia from infected fields. Aerial concentrations of sporangia, C (sporangia m^?3), were monitored using Rotorod samplers deployed at 0·5 to 3·0 m above a naturally infected cucumber canopy in two sites in central and eastern North Carolina in 2011, where disease severity ranged from 1 to 40%. Standing crop of sporangia was assessed each morning at 07·00 h EDT and ranged from 320 to 16 170 sporangia m^?2. Disease severity and height above the canopy significantly (P < 0·0001) affected C with mean concentration (C_m) being high at moderate disease. Values of C_m decreased rapidly with canopy height and at a height of 2·0 m, C_m was only 7% of values measured at 0·5 m when disease was moderate. Daily total flux (F_D) was dependent on disease severity and ranged from 5·9 to 2242·3 sporangia m^?2. The fraction of available sporangia that escaped the canopy increased from 0·028 to 0·171 as average wind speed above the canopy for periods of high C increased from 1·7 to 3·6 m s^?1. Variations of C_m and F_D with increasing disease were well described (P < 0·0001) by a log‐normal model with 15% as the threshold above which C_m and F_D decreased as disease severity increased. These results indicate that disease severity should be used to adjust sporangia escape in spore transport simulation models that are used to predict the risk of spread of cucurbit downy mildew.     

杂交籼稻新组合抗旱性鉴定评价及预测研究

为探索有效鉴定、评价及快速预测杂交籼稻新组合抗旱性的指标和方法,选择10个杂交籼稻新组合和1个对照品种为材料,采用盆栽试验进行正常水分和干旱处理。综合分析干旱胁迫条件下水稻产量及相关农艺性状的变化差异,借助产量抗旱系数、模糊数学隶属函数、抗旱性综合评价D值对杂交籼稻新组合抗旱性进行鉴定和评价,并通过主成分分析(PCAM)、相关分析(CAM)和逐步回归分析(SRM)进行抗旱指标筛选。结果表明:不同杂交籼稻新组合间抗旱能力存在显著差异,通过抗旱性综合评价D值聚类分析得出C8—沪旱7A/成旱恢30241属于强抗旱型,C_1—沪旱7A/Z4R(Si)、C_2—沪旱7A/成恢177、C_7—沪旱7A/成旱恢30248、C_9—沪旱7A/成旱恢31708和C_(10)—沪旱7A/成旱恢30218属于抗旱型,C_3—沪旱7A/成恢178、C_4—沪旱7A/成旱恢31701和C_5—沪旱7A/成旱恢31704属于中间类型,冈优725(对照品种)和C_6—沪旱7A/成旱恢31621属于不抗旱型;有效穗数、单穗实粒重和结实率的相对值可作为鉴定与评价杂交籼稻抗旱性的重要综合指标;利用逐步回归构建水稻材料抗旱性预测模型D=(245.231+1.645X_2+2.712X_4+6.494X_6+19.401X_7-2.791X_(10))×10~(-3)能准确、快速预测杂交籼稻新组合抗旱能力(其中:D为供试材料的抗旱综合评价D值,X_2、X_4、X_6、X_7和X_(10)分别为水稻干旱胁迫下有效穗数、单穗实粒数、单穗总粒重、单穗实粒重和千粒重的相对值)。     

UV protectants for Candida oleophila (strain O), a biocontrol agent of postharvest fruit diseases

This study investigated the influence of UV‐B radiation (280–320 nm) on survival of Candida oleophila strain O, an antagonist yeast that prevents postharvest diseases caused by Botrytis cinerea and Penicillium expansum on apple and pear fruits. Lethal doses (LD₅₀ and LD₉₀) were, respectively, 0·89 and 1·45 Kj m^?2 for in vitro exposure and 3·06 and 5·5 Kj m^?2 for in vivo exposure. A screening test of UV‐B protectants for strain O was also evaluated under in vitro and in vivo conditions. The in vitro results showed that sodium ascorbate (0·1% and 0·01%), riboflavin (0·1%) and uric acid (0·1% and 0·01%) were the most effective and most suitable protectants. However, only riboflavin (0·1%) and uric acid (0·1%) were effective under in vivo conditions. The efficacy obtained with strain O against P. expansum, when subjected to UV‐B radiation, was 75·0% and 49·2% for pathogen concentrations of 10⁵ and 10⁶ spores mL^?1, respectively. Adding riboflavin to strain O gave a similar efficacy (64·2%). Applying strain O together with uric acid (0·1%) was less active (47·7%). Nonetheless, its efficacy when applied with the antioxidants sodium ascorbate (71·1%) or ascorbic acid (82·5%) was the greatest. Riboflavin and uric acid were the most cost‐effective protectants, and could be included in the final formulation of strain O when applied preharvest.     

Baseline and differential sensitivity of Peronophythora litchii (lychee downy blight) to three carboxylic acid amide fungicides

Downy blight, caused by Peronophythora litchii, is an important disease of lychee (litchi) plants in China. The in vitro sensitivities of various asexual stages of P. litchii to the three carboxylic acid amide (CAA) fungicides dimethomorph, flumorph and pyrimorph were studied with four single‐sporangium isolates. None of the three fungicides affected zoospore discharge from sporangia, but they strongly inhibited mycelial growth (mean EC₅₀ values of 0·075, 0·258 and 0·115 mg L^?1, respectively); sporangial production (mean EC₅₀ values of 0·085, 0·315 and 0·150 mg L^?1, respectively); germination of cystospores (mean EC₅₀ values of 0·140, 0·150 and 0·645 mg L^?1, respectively); and germination of sporangia (mean EC₅₀ values of 0·203, 0·5 and 0·743 mg L^?1, respectively). As mycelial growth was the most sensitive stage to dimethomorph and pyrimorph, it was chosen to test baseline sensitivities to the three fungicides. In 2007, from 131 isolates collected in Fujian, Guangdong and Guangxi provinces, 127, 116 and 113 isolates were used to establish baseline sensitivity for dimethomorph, flumorph and pyrimorph respectively. Isolates from different provinces exhibited similar baseline sensitivity to the same fungicide. Baseline sensitivities to dimethomorph, flumorph and pyrimorph were distributed as unimodal curves, with mean EC₅₀ values of 0·082 (± 0·01), 0·282 (± 0·047), and 0·115 (± 0·032) mg L^?1, respectively. This information will serve as a baseline for tracking future changes in sensitivities of P. litchii populations to these three CAA fungicides.     

Baseline sensitivity of natural populations and resistance of mutants of Xanthomonas oryzae pv. oryzae to a novel bactericide,zinc thiazole

During 2009–2010, a total of 323 isolates of Xanthomonas oryzae pv. oryzae were obtained from rice with symptoms of bacterial leaf blight (BLB) in four provinces (Zhejiang, Jiangsu, Anhui and Hubei) in China. These isolates were tested for baseline sensitivity to zinc thiazole, a novel bactericide with strong antibacterial activity against Xanthomonas. The sampled pathogenic population had similar sensitivity to zinc thiazole (0·1–16·8 mg L^?1) in all four regions and over the whole two‐year study period. The baseline sensitivity was distributed as a unimodal curve with a mean EC₅₀ value of 6·79 ± 1·61 mg L^?1. The risk of mutation to resistance of zinc thiazole in X. oryzae pv. oryzae was further evaluated in vitro and in vivo. Twelve zinc thiazole‐resistant mutants were obtained through ultraviolet (UV) irradiation, culturing on zinc thiazole‐amended nutrient agar (NA) plates, and culturing on zinc thiazole‐treated rice plants. These zinc thiazole‐resistant mutants had resistance factors (RF = EC₅₀ value of a mutant / EC₅₀ value of the wildtype parent of this mutant) of 12·4 to 186·1 with a mean RF value of 44·1. Mutants obtained via UV irradiation, culturing on NA plates and culturing on rice plants had mean RF values of 51·8, 24·5 and 14·4, respectively. All mutants showed decreases in resistance to zinc thiazole after 20 successive transfers on bactericide‐free media or 10 successive inoculation–reisolations on bactericide‐free rice plants. No significant difference was found in bacterial growth and sensitivity to bismerthiazol between zinc thiazole‐resistant mutants and their parents. However, a significant decrease was observed in the pathogenicity of zinc thiazole‐resistant mutants compared with their parents, especially for mutants obtained via UV irradiation.     

轮作休耕模式对土壤细菌群落的影响

以黄土高原半干旱区9种轮作休耕模式土壤为研究对象,分别设残膜覆盖（T₂1）、伏天深耕（T₂2）、施牛羊粪（7 500 kg·hm^-2）+深翻耕（30 cm）+连续休耕3 a（T₂3）、施牛羊粪（7 500 kg·hm^-2）+深翻耕（30 cm）+连续种植豌豆3 a并在盛花期翻压还田（T₂4）、施牛羊粪（7 500 kg·hm^-2）+深翻耕（30 cm）+连续3 a种植豌豆（每kg豌豆种子拌根瘤菌粉8.5 g）并在盛花期翻压还田（T₂5）、玉米秸秆粉碎（还田量7 500 kg·hm^-2）+施牛羊粪（7 500 kg·hm^-2）+深翻耕（30 cm）+连续休耕3 a（T₂6）、玉米秸秆粉碎（还田量7 500 kg·hm^-2）+施牛羊粪（7 500 kg·hm^-2）+深翻耕（30 cm）+连续3 a种植毛苕子并在盛花期翻压还田（T₂7）、玉米秸秆粉碎（还田量7 500 kg·hm^-2）+施牛羊粪（7 500 kg·hm^-2）+深翻耕（30 cm）+连续3 a种植毛苕子并在盛花期翻压还田（T₂8）等8个处理,以连续休耕3 a为对照（CK）,采用16S rRNA扩增子测序技术对土壤细菌群落组成、丰度和多样性等特征进行研究,分析了土壤养分与细菌种群多样性的相关关系。结果表明,9种休耕轮作模式下,土壤细菌群落中相对丰度较高的前5位菌门依次为变形菌门（Proteobacteria）（26.33%～37.33%）、酸杆菌门（Acidobacteria）（17.93%～21.43%）、放线菌门（Actinobacteria）（12.42%～19.41%）、芽单胞菌门（Gemmatimonadetes）（6.98%～10.99%）和绿弯菌门（Chloroflexi）（6.48%～10.50%）。其中T₂1、T₂5处理显著升高了硝化螺旋菌门的相对丰度（分别为50.04%和42.44%）,T₂2处理显著提高了变形菌门和拟杆菌门的相对丰度（分别为31.51%和159.96%）,T₂6、T₂7处理显著增加了芽单胞菌门的相对丰度（分别为28.36%和28.33%）,T₂8处理显著增高了奇古菌门的相对丰度（519.79%）。不同轮作休耕模式的土壤全氮含量均高于CK。细菌群落丰富度指数除T₂7处理外,其余处理均高于CK。冗余分析和Monte Carlo置换检验结果显示,土壤全氮、全钾、速效氮含量对土壤细菌群落结构影响显著（P＜0.05）。     

Implications of bacterial contaminated seed lots and endophytic colonization by Pseudomonas fuscovaginae on rice establishment

The economic impact of seedborne bacterial diseases on rice production provides a major motivation for research on seed health. This paper reports on the endophytic growth of a rifampicin‐marked strain of the seedborne rice pathogen Pseudomonas fuscovaginae. The bacterium was found in most tested seeds indicating that, even without visible discolouration, seed transmission is possible. Crushed discoloured seeds contained more bacterial cells than did non‐crushed discoloured seeds. These bacteria were released during seed soaking, contaminating clean seed and lowering seed germination. Cells of a rifampicin‐resistant strain of P. fuscovaginae, which had been inoculated onto rice seeds, were subsequently recovered from different growth stages and from different rice tissues, thereby indicating endophytic colonization. These results have implications for seedling establishment, as symptomless seeds do not assure disease‐free seeds, and the presence of seedborne bacteria results in poor germination and poor seedling establishment. Elimination of seedborne bacteria by soaking in sodium hypochlorite can increase seed germination. This could be used in developing control strategies, and, if practised regularly, reduce entry of seedborne disease‐causing organisms into crops, resulting in lower disease pressure.     

Structure–biological activity relationships in triterpenic saponins: the relative activity of protobassic acid and its derivatives against plant pathogenic fungi

BACKGROUND: Triterpenic saponins from Sapindus mukorossi Gaertn. and Diploknema butyracea JF Gmelin were evaluated for in vitro antifungal activity against four phytopathogenic fungi. The study of the structure–antifungal activity relationships of protobassic acid saponins was widened by including semi‐synthetic derivatives. RESULTS: Diploknema butyracea saponins exhibited significant antifungal activity against three fungi (ED₅₀ 230–455 µg mL^?1), whereas S. mukorossi saponin was effective against two fungi (ED₅₀ 181–407 µg mL^?1). The n‐butanol extract after preparative HPLC separation provided two saponins from D. butyracea saponin mixture: 3‐O‐[β‐D ‐glucopyarnosyl‐β‐D ‐glucopyranosyl]‐16‐α‐hydroxyprotobassic acid‐28‐O‐[arabinopyranosyl‐glucopyranosyl‐xylopyranosyl]‐arabinopyranoside (MI‐I), and 3‐O‐β‐D ‐glucopyranosyl‐glucopyranosyl‐glucopyranosyl‐16‐α‐hydroxyprotobassic acid‐28‐O‐[arabinopyranosyl‐xylopyranosyl‐arabinopyranosyl]‐apiofuranoside (MI‐III). The single saponin extracted from S. mukorossi saponin mixture was identified as 3‐O‐[O‐acetyl‐β‐D ‐xylopyranosyl‐β‐D ‐arabinopyranosyl‐β‐D ‐rhamnopyranosyl] hederagenin‐28‐O[β‐D ‐glucopyranosyl‐β‐D ‐glucopyranosyl‐β‐D ‐rhamnopyranosyl] ester (SM‐I). Monodesmosides resulting from the partial degradation of hederagenin and hydroxyprotobassic acid bisdesmosides exhibited significant reduction in antifungal effect. Further removal of sugar moiety yielded complete loss in activity. The antifungal activity of the triterpenic saponins was associated with their aglycone moieties, and esterification of the hydroxyl group led to change in antifungal activity. CONCLUSION: Sapindus mukorossi saponin, which is effective against Rhizoctonia bataticola (Taub.) Briton Jones and Sclerotium rolfsii Sacc., can be exploited for the development of a natural fungicide. A sugar moiety is a prerequisite for the antifungal activity of triterpenic saponin. Copyright © 2010 Society of Chemical Industry     

In vivo pharmacokinetics of pyribenzoxim in rats

Pyribenzoxim, benzophenone O‐[2,6‐bis(4,6‐dimethoxypyrimidin‐2‐yloxy)benzoyl]oxime, is a new post‐emergence herbicide providing broad‐spectrum weed control in rice fields. [¹⁴C]Pyribenzoxim was used to study the pharmacokinetics of the compound after oral administration of a dose of 1000 mg kg^?1 to male Sprague–Dawley rats. The material balance ranged from 97.3 to 99.7% of the administered dose and urinary and fecal recovery accounted for 97.1%, with the majority of radioactivity recovered in feces (88.6%) by 168 h after treatment. Elimination as volatile products or as carbon dioxide was negligible. The following values were obtained for the compound in the blood: AUC_0–168h, 28400 µg equiv h g^?1; T_max, 12 h; C_max, 372 µg equiv g^?1; half‐life, 53 h. Radioactivity in tissue decreased from 96.1% of applied radiocarbon at 6 h to 0.4% at 168 h and the highest concentration of radioactivity among the tissues was observed in liver while the lowest residues were found in brain. The elimination half‐lives of radioactivity from tissues was in the range of 7 to 77 h and T_max values of 12, 24 and 12 h were observed for blood, liver and kidney, respectively. Except for that in the digestive tract, the tissue‐to‐blood ratio (TBR) was highest in the liver. © 2001 Society of Chemical Industry     

The role of pendimethalin in the integrated management of propanil-resistant Echinochloa colona in Central America

Pre-emergence activity of pendimethalin on propanil-resistant jungle rice (Echinochloa colona) was demonstrated in glasshouse trials. Both susceptible and resistant populations, collected from Costa Rica, were controlled by 1·25 kg ha^-1, the usual application rate used in the field where Rottboellia cochinchinensis is also a problem. When applied post-emergence, propanil performance was improved by the addition of low doses of pendimethalin to the herbicide mixture. A propanil-resistant selection was controlled by 0·23 kg ha^-1 pendi-methalin+0·54 kg ha^-1 propanil at the one-to-two leaf stage, and 0·23 kg ha^-1 pendimethalin+1·08 kg propanil at the three-to-four leaf stage compared to 1·08 kg and 2·16 kg ha^-1 respectively when propanil was applied alone. This suggests that pendimethalin improves post-emergence control in the field compared to the standard propanil treatment and can provide residual pre-emergence control of late-germinating individuals, so reducing the propanil selection pressure. For effective jungle rice control, growers apply propanil (3·84 kg ha^-1) at 10 and 20 days after planting (DAP) followed by one application of fenoxaprop-P-ethyl (0·045 kg ha^-1) at 35 DAP. Field experiments, conducted in dry-seeded upland rice in southern Costa Rica, demonstrated that under high jungle-rice population pressure, one application of pendimethalin at 1·5 kg ha^-1 provided an effective replacement for propanil, resulting in reduced weed-control costs. ©1997 SCI     

超高效液相色谱-串联质谱法检测水稻及稻田环境中速灭威的残留

建立了超高效液相色谱-串联质谱(UPLC-MS/MS)检测稻田土壤、田水、水稻植株、谷壳和糙米样品中速灭威残留的分析方法。样品经乙腈提取及盐析处理后,用乙二胺-N-丙基(PSA)和白炭黑(SiO2·nH2O)净化,UPLC-MS/MS 多离子反应监测模式下测定。结果表明,在0.005~1 mg/L 范围内,速灭威的仪器响应值与进样质量浓度间呈良好线性关系,相关系数(r)大于0.98。当添加水平为0.01~5 mg/kg(田水样品中为0.005~1 mg/L)时,速灭威在不同样品基质中的平均回收率为76.7% ~107.8%,相对标准偏差(RSD)为1.7% ~8.5%,最小检出量(LOD)均为2.0×10-13g,最低检测浓度(LOQ) 除在田水样品中为0.005 mg/L外,其余均为0.01 mg/kg。当按推荐剂量的1.5倍(有效成分45 g/hm2)分别施药2次和3次后,采用所建方法测得距最后一次施药10、14和21 d采收的糙米样品中速灭威的最终残留量均为未检出(低于0.01 mg/kg)。     

Pathogenicity and host‐parasite relationships of the root‐knot nematode Meloidogyne incognita on celery

Pathogenicity and host‐parasite relationships in root‐knot disease of celery (Apium graveolens ) caused by Meloidogyne incognita race 1 were studied under glasshouse conditions. Naturally and artificially infected celery cv. D’elne plants showed severe yellowing and stunting, with heavily deformed and damaged root systems. Nematode‐induced mature galls were spherical and/or ellipsoidal and commonly contained more than one female, males and egg masses with eggs. Feeding sites were characterized by the development of giant cells that contained granular cytoplasm and many hypertrophied nuclei. The cytoplasm of giant cells was aggregated along their thickened cell walls and consequently the vascular tissues within galls appeared disrupted and disorganized. The relationship between initial nematode population density (Pi) and growth of celery plants was tested in glasshouse experiments with inoculum levels that varied from 0 to 512 eggs and second‐stage juveniles (J₂) mL^?1 soil. Seinhorst's model y = m + (1 – m)z^P–T was fitted to height and top fresh weight data of the inoculated and control plants. The tolerance limit with respect to plant height and fresh top weight of celery to M. incognita race 1 was estimated as 0·15 eggs and J₂ mL^?1 soil. The minimum relative values (m) for plant height and top fresh weight were 0·37 and 0·35, respectively, at Pi ≥ 16 eggs and J₂ mL^?1 soil. The maximum nematode reproduction rate (Pf/Pi) was 407·6 at an initial population density (Pi) of 4 eggs and J₂ mL^?1 soil.     

Fumigant Toxicity of Citruspeel Oils against Adult and Immature Stages of Storage Insect Pests

The biological action of citruspeel oils was shown to depend on a strong fumigant action. Bioassays conducted in air-tight glass chambers showed that all the six citrus oils tested had vapour toxicity towards adults of Callosobruchus maculatus F., Sitophilus zeamais Motsch. and Dermestes maculatus Deg. The 24-h LC₅₀ value of limepeel oil (a typical citrus oil) vapour against C. Maculatus was 7·99 μl litre⁻¹ which made it 1·5 and 1·6 times less toxic against the smaller S. zeamais and the larger D. maculatus adult insects. When immature stages were fumigated, limepeel oil vapour had 24-h LC₅₀ values of 7·8 and 21·5 μl litre⁻¹ against eggs of C. maculatus and D. maculatus respectively, and 9·1, 17·8 and 23·1, 23·9 μl litre⁻¹ against early larvae, pupae of C. maculatus and late larvae, pupae of D. maculatus respectively. X-ray studies showed that fumigated C. maculatus larvae within cowpea grains died immediately without further development. The bioactivities of five other citruspeel oils were similar to that of limepeel oil. Bioassays showed that sorption of citruspeel oil fumes occurred in the presence of grains or strips of dried fish, and that this tended to reduce the amount available for fumigant action outside the materials. The problems presented by sorption may hinder the development of citrus oils into practical fumigants for large-scale treatments of stored commodities.     

Development and validation of a set of standard area diagrams to aid in estimation of spot blotch severity on wheat leaves

This study aimed to develop and validate a standard area diagram (SAD) set to quantify the severity of spot blotch, caused by Bipolaris sorokiniana, on wheat leaves. The proposed SAD set includes images of leaves with 11 distinct disease severities (0·1, 1, 5, 10, 20, 30, 40, 50, 60, 70 and 83%). The SAD set was validated by 12 raters without experience in evaluating plant disease. Lin's concordance correlation analysis of estimated versus actual disease severity (based on image analysis) showed that precision and accuracy improved for all raters using the SAD set in contrast to assessments made without it. The SAD set improved accuracy (coefficient of bias, C_b = 0·88 and 0·99, without and with the SAD set, respectively) and agreement (Lin's concordance correlation coefficient, ρ_c = 0·81 and 0·96 without and with the SAD set, respectively) of the estimates of severity. The severity estimates were also more reliable when using the SAD set (coefficient of determination, R² = 0·76 unaided and R² = 0·92 with the SAD set, and intra‐class correlation ρ = 0·79 without the SAD set and ρ = 0·95 using the SAD set). The SAD set proposed in this study will improve the accuracy and reliability of estimates of spot blotch severity on wheat leaves.     

Presence of Muscarinic Acetylcholine Receptors in the Cattle Tick Boophilus microplus and in Epithelial Tissue Culture Cells of Chironomus tentans

A muscarinic acetylcholine receptor (mAChR) has been demonstrated and partially characterized in larvae of the cattle tick Boophilus microplus. Its properties are compared with mAChR from an epithelial cell line from the dipteran insect Chironomus tentans. Competition studies with cholinergic ligands of different specificity revealed the muscarinic nature of the cholinergic receptors investigated in both species. In homogenates from tick larvae, specific binding sites for [³H]quinuclidinyl benzilate (QNB) with high affinity (1·2±(0·13) nM ; B_max 22·5 pmol mg protein⁻¹) were detected that do not bind nicotinic compounds specifically. The estimated IC₅₀ values for nicotine, imidacloprid and α-bungarotoxin were all in the mM range. Additionally, with tick larvae, high-affinity nicotinic binding sites were detected with [³H]nicotine which could be displaced by high concentrations of imidacloprid or QNB. The estimated IC₅₀ values for nicotine, α-bungarotoxin, imidacloprid and QNB were 43(±8) nM , 0·8(±0·2) μM , 2·8(±0·6) μM and 78(±1·9) μM , respectively. With homogenates of the non-neuronal insect cell line from C. tentans, only high-affinity binding sites for [³H]QNB were found. Muscarinic antagonists selectively displaced [³H]quinuclidinyl benzilate (QNB) binding to tick larvae homogenates. The mAChR of B. microplus preferred pirenzepine (IC₅₀ 2·13(±1·02) μM ) among different subtype-specific mAChR antagonists (4-DAMP had IC₅₀ 49·9(±9·13) μM and methoctramine had IC₅₀ 121(±14·2) μM ) indicating a type of binding site similar to the vertebrate M1 mAChR subtype. The tick muscarinic receptor seems to be a G-protein-coupled receptor, as concluded from the 4·8-fold reduction in receptor affinity for binding of the muscarinic agonist oxotremorine M upon treatment with the non-hydrolysable GTP-analogue γ-S-GTP. Binding data for the agonists oxotremorine M (IC₅₀ 71·3(±19·6) μM ) and carbachol (IC₅₀ 253(±87·1) μM ) parallel the biological efficacy of these compounds, in that, while oxotremorine M showed some activity against ticks, carbachol was ineffective.     

Effect of electrical conductivity on survival of Phytophthora alni,P. kernoviae and P. ramorum in a simulated aquatic environment

This study investigated survival of the pathogens Phytophthora ramorum, P. alni and P. kernoviae as zoospores or sporangia in response to an important water quality parameter, electrical conductivity (EC), at its range in irrigation water reservoirs and irrigated cropping systems. Experiments with different strengths of Hoagland’s solution showed that all three pathogens survived at a broad range of EC levels for at least 3 days and were stimulated to grow and sporulate at ECs > 1·89 dS m^?1. Recovery of initial populations after a 14‐day exposure was over 20% for P. alni subsp. alni and P. kernoviae, and 61·3% and 130% for zoospores and sporangia of P. ramorum, respectively. Zoospore survival of these pathogens at ECs < 0·41 dS m^?1 was poor, barely beyond 3 days in pure water; only 0·3% (P. alni), 2·9% (P. kernoviae) and 15·1% (P. ramorum) of the initial population survived after 14 days at EC = 0·21 dS m^?1. The variation in rates of survival at different EC levels suggests that these pathogens survive better in cropping systems than in irrigation water. Containment of run‐off and reduction in EC levels may therefore be non‐chemical control options to reduce the risk of pathogen spread through natural waterways and irrigation systems.     

水稻抗白叶枯病基因Xa47(t) a的敲除及抗性鉴定

为鉴定水稻Xa47(t)a基因对白叶枯病的抗性,以水稻种质L214为材料,通过构建针对Xa47(t)a基因的Xa47(t)a-Cas9敲除载体来获得敲除突变体,利用生物信息学技术对突变的Xa47(t)a基因进行突变类型分析,同时采用实时荧光定量PCR(quantitative real-time PCR,qPCR)技术分析突变体中Xa47(t)a及病程相关基因的表达情况,并于水稻孕穗期对突变体及其野生型植株接种11株白叶枯病菌Xanthomonas oryzae pv. oryzae菌株进行抗性鉴定。结果表明,在Xa47(t)a基因的第2外显子区域进行基因编辑后成功获得25株T₀代突变体株系;测序分析发现T₀代突变体株系中有13种不同的突变体类型,其中纯合突变体有3种类型,且突变位点均在靶标位点的11位碱基处缺失1～4个A碱基;氨基酸序列分析发现大部分突变体中Xa47a编码的蛋白翻译会提前终止,由原来的803个氨基酸变为144～166个氨基酸;qPCR分析结果表明突变体中Xa47(t)a及大部分病程相关基因的表达水平显著低于野生型株系;抗性鉴...     

高毒力金龟子绿僵菌的筛选及其胞外蛋白酶产量测定

以大蜡螟Galleria mellonella L.幼虫为供试昆虫,对从果园土壤中筛选出的金龟子绿僵菌Metarhizium anisopliae的生物活性进行了测定,共获得4株高毒力菌株,其LT50值在1.85～2.56 d之间,LD50值在6.11×102～1.55×104 spores之间。利用明胶-琼脂平板分析法对筛选出的高毒力绿僵菌菌株产胞外蛋白酶的水平进行了测定,结果显示,各菌株间产酶水平差异较大,且菌株产酶水平的高低与其对大蜡螟幼虫的毒力之间存在明显的线性相关性。