Behavioral Effects and Retention of Protein Immunomarkers on Plum Curculio Conotrachelus nenuphar (Coleoptera: Curculionidae)

Protein immunomarking can be used to track the dispersal of insects in the field or identify plant–insect interactions. By marking insects with known proteins and recapturing them, their movement or host use can be quantified with Enzyme Linked Immunosorbent Assay (ELISA). Before using this technique, retention and behavioral effects of these markers should be evaluated to ensure that the insect’s natural behaviors are conserved. Here, we tested the effects of protein markers on the plum curculio, Conotrachelus nenuphar (Coleoptera: Curculionidae) using two different application methods. This weevil is native to North American and a pest of tree fruit and blueberry in the United States and causes damage resulting in near complete crop loss if left untreated. We tested the effects of marking adult C. nenuphar with two inexpensive food-based immunoprotein markers, bovine casein (cow’s milk) and chicken albumin (egg whites) on climbing distance (total cm), lateral movement (total cm), and lateral movement speed (cm/s), as well as retention time of protein immunomarkers. Neither protein immunomarker affected C. nenuphar movement or climbing, although females climbed significantly greater distances than males. ELISA assays detected 37.5–56.2% of milk protein and 56.2–59.3% of egg on the insect 7 d after application depending on application method. Our findings indicate that food-based protein immunomarkers can be used in future studies to test C. nenuphar movement within host plants without impacting behavior. The use of protein immunomarking will allow studies that will lead to behaviorally based management tactics.     

Optimization of process parameters influencing the submerged fermentation of extracellular lipases from Pseudomonas aeruginosa, candida albicans and Aspergillus flavus

The present study was aimed at optimization, production and partial purification of lipases from Pseudomonas aeruginosa, Candida albicans and Aspergillus flavus. Various nutritional and physical parameters affecting lipase production such as carbon and nitrogen supplements, pH, temperature, agitation speed and incubation time were studied. Refined sunflower oil (1% v/v) and tryptone at a pH of 6.2 favored maximum lipase production in Pseudomonas at 30 degrees C and 150 rpm, when incubated for 5 days. In C. albicans refined sunflower oil (3% v/v) and peptone resulted in maximum lipase production at pH 5.2, 30 degrees C and 150 rpm, when incubated for 5 days. In A. flavus coconut oil (3% v/v) and peptone yielded maximum lipase at pH 6.2, 37 degrees C, 200 rpm after an incubation period of 5 days. The lipases were partially purified by ammonium sulphate precipitation and dialysis. In P. aeruginosa enzyme activity of the dialyzed fraction was found to be 400 U mL-' and for C. albicans 410 U mL(-1). The dialysed lipase fraction from A. flavus demonstrated an activity of 460 U mL(-1). The apparent molecular weights of the dialyzed lipases were determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The dialyzed lipase fraction obtained from P. aeruginosa revealed molecular weights of 47, 49 and 51 kDa, whereas, lipases from C. albicans and A. flavus demonstrated 3 bands (16.5, 27 and 51 kDa) and one band (47 kDa), respectively. These extracellular lipases may find wide industrial applications.     

Effects of cultural conditions on high temperature tolerance of Lentinula edodes mycelia

The effects of several cultural conditions on high temperature tolerance of vegetative mycelia of five Lentinula edodes strains were investigated. Mycelium of longer culture age (70 days) was significantly shown high temperature tolerance compared to mycelium of shorter culture ages (14 and 30 days) for four strains, whereas SA142 that showed reverse. When the culture plates were pretreated at 30 and 33 degrees C for 48 h incubation before heat treatment (40 degrees C, 8 h), mycelia of L. edodes strains were shown high temperature tolerance. Effects of nutritional factors in BM (basic medium) for the high temperature tolerance of L. edodes strains were also investigated. While yeast extract (as nitrogen source) and starch (as carbon source) were added to BM media, as a result cultured vegetative mycelia of L. edodes strains were significantly shown high temperature tolerance against heat treatment (40 degrees C, 6 and 8 h). Further more, addition of bases (adenine + cytosine), vitamin (biotin) and organic acid (tartaric acid) to BM media, consequently vegetative mycelia of some L. edodes strains were also effective to increase high temperature tolerance.     

保健砂对文昌鸡产蛋性能及蛋壳品质的影响

为探讨保健砂在蛋鸡生产中的合理应用,将450只文昌鸡（24周龄蛋鸡）随机分为5组,每组90只。对照组饲喂基础日粮和添加剂,试验组饲喂基础日粮、添加剂和保健砂,各试验组添加比例分别为2％、4％、6％和8％。同期对照组与试验组Ⅳ的产蛋率、料蛋比差异均显著（P<0.05）,对照组与试验组Ⅰ、Ⅱ、Ⅲ的产蛋率、料蛋比差异不显著（P>0.05）,平均蛋重、蛋壳厚度以及破损率各试验组与对照组对比差异均不显著（P>0.05）。试验结果说明饲料中添加4％～6％保健砂,有助于提高文昌鸡的产蛋率、减少饲料消耗,提高养鸡生产经济     

Rates of internal blackspot bruise development in potato tubers under conditions of elevated temperatures and gas pressures

Internal blackspot bruising was produced in Russet Burbank tubers by dropping a 150 g weight 40 cm through a slotted guide tube and incubating the tubers for varying time periods at temperatures ranging from 10 to 80 C. When potatoes were bruised following 1 to 5 months storage, those tubers incubated at 10 C required 48 hr for maximum blackspot development. Increasing the incubation temperature accelerated blackspot formation with an optimum response at 36 to 40 C. At 40 C tubers reached maximum discoloration within 6 hr. Bruised tubers were also exposed to combinations of varied temperatures and varied gas pressures of 0.3 to 2.1 kg/cm² using air, O₂, CO₂, and N₂. Pressurized air slightly enhanced the temperature effect at 40 C; pressurized O₂ had no significant effect upon blackspot development, while CO₂ and N₂ inhibited blackspot formation. Blackspot bruises incurred in the field during harvest required a longer incubation period at a lower temperature than did bruises inflicted on tubers held in storage for 1 to 5 months.     

Potato clone variation in blackspot and soft rot susceptibility,redox potential,ascorbic acid,dry matter and potassium

Tubers of 11 potato clones were compared for susceptibility to blackspot and soft rot, redox potential, ascorbic acid, dry matter, and potassium content. Evaluations were made before and after 6 mo storage at 3.9C. Clonal behavior was generally consistent over 2 yr. Significant differences occurred among clones for each factor except redox potential. Blackspot and soft rot were positively correlated in 5 of 8 comparisons. Neither blackspot nor soft rot was correlated with ascorbic acid or redox potential. Blackspot was positively correlated with dry matter in the fall but not after storage and was negatively correlated with potassium. Soft rot evaluated in air or air plus 8% CO₂ was positively correlated with dry matter in only 2 of 8 comparisons. Soft rot susceptibility did not increase with storage when inoculated discs were incubated in air but did increase when the discs were incubated in air plus 8% CO₂. Ascorbic acid was negatively correlated with redox potential only before storage and decreased to varying degrees in all clones during storage.     

Bacterial soft rot potential in washed potato tubers in relation to temperatures of tubers and water during simulated commercial handling practices

The bacterial soft rot potential in potato tubers was affected by differences in temperature between tubers and suspensions ofErwinia carotovora at the time of inoculation by immersion. In general, disease severity after incubation was higher when tuber temperatures were higher as opposed to equal to or lower than that of water containing cells of the bacterium. Disease severity was also greater when tubers were stored at 20° to 23°C in contrast to 4° or 26° to 30°C for 16–18 h prior to immersion. Tubers with a pulp temperature of 8°C were more prone to bruise injury as compared with those at 18°C when struck with a pendulum type bruising instrument. When the injured tubers were inoculated and incubated, most bruises became infected with soft rot bacteria.     

Modification of ELISA by Replacing Incubation of Microtiter Plates in an Incubator with Their Shaking in PVY,PVM and PLRV Detection

ELISA (enzyme-linked immunosorbent assay) is a sensitive and reliable method of plant virus detection. It is commonly used in daily research carried out by scientific institutions and laboratories working on the certification of potato tubers. The key stage in this method is a 3–4-h-long incubation of microtiter plates with IgG and with a conjugate in an incubator at a temperature of 37 °C. The aim of the research was to replace this type of incubation process with a technique of mechanically shaking the plates using a shaker to induce a vibrating movement. Three durations of shaking, performed at room temperature, were adopted: 30, 60 and 90 min with two incubation periods at a temperature of 37 °C: 60 and 180 min which were applied at the stage of coating the IgG plates, following addition of the conjugate. The assessment was made for three dilutions of lyophilized sap from leaf of potatoes (1:10, 1:100, 1:1,000). Replacing the stages of plates incubation with IgG and conjugate at 37 °C with mechanical shaking allowed the whole process of DAS-ELISA to be reduced below 3–4 h without any significant impact on its quality. The process turned out to be equally efficient as the 3-h-long incubation in an incubator for PVY, PVM and PLRV detection by means of DAS ELISA. Applying the 90-min-long incubation on a shaker in comparison to a 3-h-long incubation in an incubator gave comparable or even slightly improved results. The reaction background, i.e. the value of absorbance for sap from healthy plants (negative control) was very low in all the combinations irrespective of the time of reading after the substrate was placed. No significant differences for this parameter were found between the combinations and times of reading. Only in the case of PLRV was a clearly visible decrease in test sensitivity found (no positive reactions) at diluted sap over 1:10. Moreover, it was observed that an increase in dilutions impacted the length of reaction. The dilution 1:10 seemed to be the most favorable (maximum 1:100 for PVY and PVM), wherein the sensitivity and pace of staining the substrate for each of the methods did not provoke any doubts regarding the reliability of the test.     

Germination of Astragalus hamosus L. and Coronilla scorpioides (L.) as influenced by temperature

Seed germination of two pasture legumes was tested under dark conditions at three alternating temperatures of 5-20, 15-20, 20-35 degrees C and constant 20 degrees C. Duration of the alternating temperatures was 12 h. The seedling counts were checked every day during four weeks. Species differed significantly in germination response to temperature. A. hamosus was more sensitive to temperature than C. scorpioides. The temperature of 20-35 degrees C was more favourable for the germination of A. hamosus than constant 20 degrees C which markedly reduced its germination. The interaction between accessions and temperatures for total germination was significant in both species. In A. hamosus, almost 50% of germination was achieved by all accessions by the first week at alternating 20-35 degrees C temperature while in C. scorpioides, average total germination was significantly low (4%) even by the 28th day. A total rate of 11 and 9% of germination were recorded by two accessions from Ouesslatia (210) and Agareb (19), respectively. A considerable variation for germination was observed among accessions of both species. Temperature requirements for seed germination of the studied species suggest why they are relatively easy or difficult to establish in the field. A. hamosus, with very small seeds required relatively high alternating temperature for germination than C. scorpioides which was insensitive to temperature and has relatively bigger seeds.     

Changes in the content of free sulphydryl groups during postharvest wheat and flour maturation and their influence on technological quality

The main aim of this study was to determine the changes in free sulphydryl content during postharvest wheat and flour maturation. The content of free sulphydryl groups was determined from wet gluten over 50 days of wheat postharvest maturation and over 14 days of flour maturation by varying incubation temperatures (30 and 37 °C) and incubation times (0, 45, 90 and 135 min). The amount of free sulphydryl groups increased during postharvest wheat and flour maturation as well as with the increase in temperature and gluten incubation time. The additional aim of this study was to find the interrelation between the content of free sulphydryl groups and selected parameters of technological quality by means of Principal Component Analysis. During wheat/flour maturation, the strengthening of protein structure was observed manifested by the increase in gluten index and Mixolab protein network weakening (C2), and changes in dough resistance as well. The incubation temperature of 37 °C affected the weakening of protein structure manifested by the decrease in gluten index and resistance, and increase in extensibility. Strong differentiation of examined varieties in terms of selected quality indicators was observed upon pre-defined maturation period, probably due to the expression of their intrinsic properties.     

Thermal Biology of Liorhyssus hyalinus (Hemiptera: Rhopalidae) and Nysius simulans (Hemiptera: Lygaeidae), Fed on the Milky Stage of Maize Grains

When quinoa, Chenopodium quinoa Willd., is cultivated in South America outside of its Andean origin, the heteropterans Liorhyssus hyalinus (Fabricius) and Nysius simulans Stål may emerge as important pests. Here we studied the development and reproduction of both species at different constant temperatures in the laboratory. Egg and nymphal development were investigated at 18, 22, 26, 30, 34, and 36°C. For both species, egg incubation time significantly decreased as the temperature increased. Nymphs did not successfully develop at 18°C and the total nymphal time significantly decreased as the temperature increased from 22 to 36°C. Based on a linear day-degree (DD) model, the lower developmental threshold (LDT) temperatures for eggs and nymphs were estimated to be 16.0 and 17.9°C for L. hyalinus, and 16.1 and 19.7°C for N. simulans, respectively. Thermal requirements for egg and nymphal development were 68.6 and 114.8 DD for L. hyalinus, and 77.7 and 190.3 DD for N. simulans, respectively. Reproduction and adult longevity were studied at 22, 26, 30, and 34°C. For both species preoviposition time decreased as temperature increased, and the oviposition period was longest at 26°C. The highest fecundity and egg viability were observed at 30°C, whereas longevities were higher at 22–26°C than at 30–34°C. As the lowest tested temperatures were not suitable to both heteropterans and 30°C was found to be the optimal temperature for development and reproduction, peak densities are expected in warm areas and seasons.     

Changes in the content of free sulphydryl groups during postharvest wheat and flour maturation and their influence on technological quality

The main aim of this study was to determine the changes in free sulphydryl content during postharvest wheat and flour maturation. The content of free sulphydryl groups was determined from wet gluten over 50 days of wheat postharvest maturation and over 14 days of flour maturation by varying incubation temperatures (30 and 37 °C) and incubation times (0, 45, 90 and 135 min). The amount of free sulphydryl groups increased during postharvest wheat and flour maturation as well as with the increase in temperature and gluten incubation time. The additional aim of this study was to find the interrelation between the content of free sulphydryl groups and selected parameters of technological quality by means of Principal Component Analysis. During wheat/flour maturation, the strengthening of protein structure was observed manifested by the increase in gluten index and Mixolab protein network weakening (C2), and changes in dough resistance as well. The incubation temperature of 37 °C affected the weakening of protein structure manifested by the decrease in gluten index and resistance, and increase in extensibility. Strong differentiation of examined varieties in terms of selected quality indicators was observed upon pre-defined maturation period, probably due to the expression of their intrinsic properties.     

稻鱼生态系统对水稻产量及其构成因素的影响

为研究不同生态综合系统对水稻产量及构成因素的影响,以稻鱼生态系统(RF)为例,选择稻鱼鸡和稻鳅两种模式,加入细绿萍,共设5个处理:稻鱼鸡(A)、稻鱼鸡萍(B)、稻鳅(C)、稻鳅萍(D)、常规稻作(CK),比较不同生态综合系统成熟期水稻干物质重的差异.结果表明,稻鱼生态系统显著提高了水稻产量,平均增产12.13%(6.93%~22.23%),且RF中含萍模式的增产量大于不含萍模式,平均增产4.55%;稻鳅模式的增产量大于稻鱼鸡模式,平均增产31.54%.单株实粒数、有效穗数和结实率呈现不同程度的增加;从成熟期的茎、叶、穗的干物质重来说,稻鱼生态系统高于常规处理,特别是穗重平均提高了13.76%,说明稻鱼生态系统有利于水稻增产增收.     

Safety of human therapeutic morphine vaccine employing Lohmann specific pathogen free eggs

One of the sensitive and standard tests to control the safety of a vaccine is the inoculation of such vaccine to the air pocket of Lohmann specific pathogen free eggs. The aim of this study is to control the safety of morphine vaccine. This study reveals the safety of morphine vaccine by employing Lohmann specific pathogen free embryo eggs. The changeable parameters in this test were: weight of eggs, safety of eggs embryo, vaccine concentration, normal saline and temperature of the incubator. To study, the safety of morphine vaccine, we used 30 eggs (after controlling the safety of eggs and their embryos) which were divided into two groups of control (15 eggs) and test (15 eggs). After weighing the eggs, the eggs under experiment were inoculated with morphine vaccine and the control group was inoculated with physiological solution. Both groups were incubated and weight of the eggs and chickens were determined accordingly. The eggs of each group were controlled by their weights showing healthy, normal growth and evolution. The comparison between the weights of control and experimental groups did not show any significant changes. Exactly growth and evolution of each group were found equally to be balancing for three weeks after injection. Finally all eggs were observed to be safe, alive and in evolutionary form. By comparing the growth and evolution amongst each egg in the group under experiment, after injection, the eggs did not show any adverse reaction after inoculation with therapeutic human morphine vaccine.     

黄秋葵叶粉对海兰褐蛋鸡部分生产性能及蛋品质的影响

研究了黄秋葵叶粉对产蛋高峰期海兰褐蛋鸡部分生产性能和蛋品质的影响，旨在找出最适宜添加量，为进一步开发新型天然饲料添加剂奠定基础。试验组黄秋葵叶粉添加量分别为3％（T3）、4％（T4）、5％（T5）、6％（T6），对照组为不添加黄秋葵叶粉组（0％，T0）。试验共进行12周，预饲期1周。结果表明：试验组蛋重较对照组均有不同程度的提高，T3、T4组的产蛋率、产蛋量比对照组有所提高，T4、T5组采食量和T4组的料蛋比较对照组有所降低；各试验组在试验鸡蛋黄的着色效果均较对照组有显著提高（p＜0.05）；各试验组哈     

Changes in peroxidase and phenols activity in apple fruit inoculated with antagonistic Pseudomonas fluorescens isolates and Botrytis mali

The biocontrol activity of three isolates of Pseudomonas fluorescens against gray mold of apple fruit caused by Botrytis mali and their ability to induce biochemical defense response in apple tissue were investigated. Apple fruit (Malus domestica) wounds were inoculated with 20 microL bacterial suspension (10(8) CFU mL(-1)) of Pseudomonas fluorescens followed 24 h later by 20 microL of conidial suspension of B. mali (10(5) conidia mL(-1)). The apples were then incubated at 20 degrees C for 11 days. Lesion diameters were evaluated 6 and 10 days after pathogen inoculation. In addition to controlling gray mold, these three isolates of P. fluorescens caused increase in peroxidase activities that reached maximum levels 2-6 days after pathogen inoculation. Phenolic accumulation was increased in apple fruit treated with antagonists and inoculated with B. mali and exhibited the highest level 6-8 days after treatment. The ability of P. fluorescens to increase activities of peroxidase and levels of phenol compounds maybe one of mechanism responsible its biocontrol activity.     

TDZ和BA对诱导大豆胚轴植株再生的影响

大豆未成熟种子和萌动种子胚轴在含有ＴＤＺ和ＢＡ的培养基上,通过器官发生,可获得再生株。培养基中同时加入ＴＤＺ和ＢＡ〈能够增加不定芽的形成和再生株的数量,其中以２．０ｍｇ／ＬＢＡ＋０．７５ｍｇ／ＬＴＤＡ处理再生效率最高。诱导培养基中适当增加ＮＡＡ浓度有利于不定芽的分化。     

Methods to increase the starch content of potato tuber callus tissue

Summary Attempts were made to increase the starch content of callus tissue from tubers of cvs Lemhi Russet and Russet Burbank by varying growth regulator concentration, sugar source, sugar concentration, temperature and callus shape. Starch content was always higher in media containing no growth regulators. Callus spread over the media contained more starch than unbroken rectangular or cube-shaped callus. Sucrose was superior to glucose and fructose for starch formation. A sucrose concentration of 20 % produced more starch than lower concentrations. Callus growth decreased as the sucrose concentration increased. ‘Lemhi Russet’ callus incubated with 8 % sucrose at 20, 25 and 30 °C contained 1.2, 0.4 and 0.8 % starch respectively, which was much higher than in ‘Russet Burbank’ callus. With varying amounts of mannitol as an osmoticum in 8 % sucrose, the most starch was found in the medium containing 5 % mannitol. Research Paper 92B2 of the Idaho Agricultural Experiment Station     

Latent infection of Botrytis cinerea in rose flowers and combined chemical and physiological control of the disease

A high percentage of rose flowers collected during winter from greenhouses around Israel developed grey mould disease when incubated in the laboratory in favourable conditions. Symptomless flowers of various cultivars were incubated in a humidity chamber after surface disinfection. More than 50% of the symptomless flowers yielded the pathogen Botrytis cinerea Pers., thus showing the presence of latent infection which probably occurred 3–8 days earlier in the commercial greenhouses. Initially, the symptoms which occurred on affected petals were mainly restricted lesions. However, when flowers were incubated under favourable conditions, necrosis developed and spread. Chemical control of grey mould was tested on cut rose flowers. Partial control of the disease was correlated with reduction in necrosis and a relative increase in restricted lesions (<2 mm in diameter). Of the 18 fungicides which were tested, the following were the most efficient during incubation at 20°C: metomeclan, dichlofluanid, myclozolin, N-phenylcarbamate plus carbendazim (NPC + MBC), polyoxin D, prochloraz and iprodione. When incubated at 10°C, the most effective fungicides against grey mould of rose flowers were NPC + MBC, polyoxin D, chlorothalonil, the detergent Tween 20, and the inhibitor of ethylene activity, silver thiosulphate (STS). Diphenylamine was more effective at 4°C than at 10° or 20°C. STS in combination with each of the fungicides chlorothalonil, polyoxin B, myclozolin or merpan was additively more effective than STS or each of the respective fungicides alone.