营养对不同地理种群周氏啮小蜂生物学特性的影响

【目的】研究补充营养对周氏啮小蜂海南本土地理种群和北京地理种群的生物学参数是否有差异。【方法】设置不同的营养(10%蜂蜜、10%蔗糖、10%葡萄糖、水、空白)分别饲喂周氏啮小蜂,统计2个地理种群的寄生率、羽化率、出蜂量、寿命。【结果】2个地理种群的周氏啮小蜂羽化后不久即可进行交配,补充10%蜂蜜、10%蔗糖、10%葡萄糖能显著提高2个地理种群的寄生率、羽化率、出蜂量、雌雄蜂寿命,补充不同的营养对两个地理种群的雌雄比、发育历期的影响差异不显著;饲喂10%蜂蜜的海南地理种群寄生率最高(84.60%),饲喂10%葡萄糖的海南地理种群羽化率最高(97.47%),饲喂10%葡萄糖的海南地理种群出蜂量最高(60.03头),饲喂10%葡萄糖的海南地理种群雌、雄蜂的寿命均为最高,分别为9.13和8.53d。营养和地理种群的交互作用对周氏啮小蜂雄蜂的寄生率、羽化率、出蜂量、雌雄比、雌雄蜂寿命具有显著影响,而对周氏啮小蜂的发育历期无显著影响。【结论】补充营养可不同程度促进周氏啮小蜂生殖力、寿命及寄生率,对寄生蜂繁育和利用其进行生物防治具有一定的意义。     

白蛾周氏啮小蜂产卵器感器的形态和超微结构

采用扫描电镜观察了白蛾周氏啮小蜂Chouioia cunea Yang产卵器.结果表明,在该蜂的产卵器上共发现了8种感器,分别为毛状感器、栓锥感器、浅凹状感器、乳状感器、钟形感器、栉齿状感器、短锥感器和B(o)hm氏鬃毛.对产卵器感觉器的形态、分布进行了描述.     

利用生命表评价白蛾周氏啮小蜂对美国白蛾的控制作用

为了研究白蛾周氏啮小蜂对侵入性害虫-美国白蛾的控制作用,在山东省烟台市美国白蛾发生区,选择美国白蛾发生程度中等的两块样地,一块样地作为释放白蛾周氏啮小蜂防治美国白蛾的防治区,另一块样地作为对照区。应用生命表技术,连续2年共4代对美国白蛾种群数量进行定点观察。防治区每代美国白蛾在化蛹初期和化蛹盛期各放蜂1次,即每代美国白蛾共放蜂2次。将2年的第1代和第2代观测数据分别综合平均,制成4个生命表。结果表明,在放蜂防治区,第1代和第2代美国白蛾的种群趋势指数（I）分别为0.29和0.14,说明下代美国白蛾种群数量将急剧下降;放蜂区的I值显著小于对照区的8.74和4.48。生命表研究结果清楚地表明,通过人工释放白蛾周氏啮小蜂能够达到良好的控制美国白蛾的效果。     

Effect of different nutrition on the main biological characteristics of Chouioia cunea

补充10％果糖水、10％蔗糖水、10％葡萄糖水、10％蜂蜜水后,均显著提高了亲代蜂的寿命,其中10％的果糖水效果最好,比对照长2.13d;补充10％果糖水和10％葡萄糖水均能显著延长子代蜂的寿命,并显著提高了子代蜂的出蜂量,其中10％的果糖水处理效果最好.发生寄生行为的小蜂,其寿命明显短于未发生寄生行为的小蜂.     

白蛾周氏啮小蜂滞育诱导及滞育后发育

本研究针对人工繁育白蛾周氏啮小蜂Chouioia cunea Yang过程中出现的小蜂滞育现象, 对其滞育诱导的光周期反应及敏感光照虫态进行了调查。结果表明: 沈阳地区的白蛾周氏啮小蜂属长日照型昆虫, 以老熟幼虫进入滞育状态, 但在不同的温度条件下诱导滞育的临界光周期不同, 在18℃时诱导滞育的临界光周期处于13L∶11D和14L∶10D之间; 在21℃和24℃时诱导滞育的临界光周期变短, 处于12L∶12D和13L∶11D之间。白蛾周氏啮小蜂滞育诱导的敏感光照虫态为幼虫期, 且以幼虫的后期最为敏感, 但整个幼虫期接受短光照对滞育的形成更为有利。通过观察白蛾周氏啮小蜂滞育后在18℃, 21℃, 24℃和30℃的恒温条件下的发育历期, 由最小二乘法计算出白蛾周氏啮小蜂老熟幼虫滞育后发育起点温度和有效积温分别为14.60±0.31℃和209.38±8.72日·度。这些结果可为进一步研究白蛾周氏啮小蜂的种蜂长期保存技术和指导商品蜂生产, 正确把握放蜂时机提供理论依据。     

周氏啮小蜂(海南种群)对椰子织蛾的寄生功能反应

【目的】周氏啮小蜂(海南种群)是寄生椰子织蛾蛹期的本土天敌。研究周氏啮小蜂(海南种群)对椰子织蛾的功能反应,能为该蜂室内扩繁和室外控害利用提供基础。【方法】在室内观察周氏啮小蜂(海南种群)寄生行为基础上,研究了营养(10%蜂蜜)、寄主蛹(初、中、晚)期对周氏啮小蜂(海南种群)寄生功能反应的影响。【结果】镜下观察1头周氏啮小蜂(海南种群)可寄生多头椰子织蛾蛹,寄生量随寄主密度的增大而增加,但当寄主密度达到一定程度后,其增长率随寄主密度的增大逐渐减缓,属于HollingⅡ功能反应。在无营养条件下,周氏啮小蜂(海南种群)对椰子织蛾初、中、晚期蛹的理论最大寄生量分别为1.5、1.4、1.2头·d~(-1);在10%蜂蜜饲喂条件下,周氏啮小蜂(海南种群)对椰子织蛾初、中、晚期蛹的理论最大寄生量分别为1.8、1.7、1.3头·d~(-1)。【结论】HollingⅡ型拟合结果表明,椰子织蛾初期蛹是周氏啮小蜂(海南种群)寄生的最佳时期,补充营养能够提高椰子织蛾的寄生力。     

白蛾周氏啮小蜂气味结合蛋白OBP1与寄主挥发物的分子对接研究

【目的】白蛾周氏啮小蜂为重大入侵害虫美国白蛾的主要天敌。本课题组前期通过转录组测序技术筛选出8个主要在白蛾周氏啮小蜂雌性触角中表达的气味结合蛋白OBPs。然而目前,对这些OBPs的具体结构和功能仍不清楚。因此,选取一个在雌性周氏啮小蜂触角特异表达的气味结合蛋白OBP1,通过分子对接技术模拟寄主挥发物与OBP1的结合情况。【方法】通过Swiss-Model对白蛾周氏啮小蜂气味结合蛋白CcOBP1进行同源建模,获得该蛋白的三维结构。从Pubchem下载γ-丁内酯、邻苯二甲酸二甲酯和萘等11种小分子的三维结构。用Schrodinger Suites 2015-2中的maestro10.2软件进行分子对接。【结果】在11种挥发物中,有3种与CcOBP1结合特性较好的小分子物质,分别是γ-丁内酯、邻苯二甲酸二甲酯和萘。【结论】白蛾周氏啮小蜂气味结合蛋白CcOBP1与γ-丁内酯、邻苯二甲酸二甲酯和萘结合特性较好,CcOBP1的功能可能与白蛾周氏啮小蜂的趋避效应相关,该结果初步探明了白蛾周氏啮小蜂OBP1的功能,可为白蛾周氏啮小蜂嗅觉分子机制的研究积累数据。     

多杀菌素对天敌昆虫白蛾周氏啮小蜂的毒力评估

[目的] 生物防治与化学防治相结合是害虫综合防治的重要策略,蛹寄生蜂白蛾周氏啮小蜂是一种重要的天敌昆虫,在美国白蛾的生物防控中起关键作用。本研究旨在评估农林生产中常用的广谱、低毒杀虫剂多杀菌素对白蛾周氏啮小蜂的安全性。[方法] 采用药膜法检测多杀菌素对小蜂的毒力,并检测亚致死浓度多杀菌素处理后,小蜂体内解毒酶活性的变化。[结果] 多杀菌素对小蜂无短期（1 h）触杀作用,但随施药时间延长,其对小蜂的毒杀作用增强,25 mg·L^-1多杀菌素施药2 h后,可诱导约18.67%小蜂个体死亡。施药4 h后,不同浓度多杀菌素对小蜂的毒杀作用差异最明显,低浓度（5 mg·L^-1）多杀菌素可诱导11.33%小蜂死亡,而中、高浓度多杀菌素（≥ 15 mg·L^-1）可诱导77.33%~88.00%个体死亡。施药6 h后,LC₅₀值达7.44 mg·L^-1,安全系数为0.31,属高风险性农药。此外,亚致死浓度多杀菌素可明显诱导小蜂羧酸酯酶活性提高,对乙酰胆碱酯酶与谷胱甘肽-S-转移酶活性具有剂量效应。[结论] 多杀菌素对白蛾周氏啮小蜂的毒力作用较强,在美国白蛾生防区需慎用。     

白蛾周氏啮小蜂毒液对美国白蛾蛹血细胞免疫的影响

【目的】明确白蛾周氏啮小蜂Chouioia cunea毒液对其寄主美国白蛾Hyphantria cunea蛹细胞免疫的影响。【方法】采用Na_2-EDTA分离美国白蛾蛹颗粒细胞,尼龙毛法分离浆血细胞,再利用细胞离体培养法,测评了白蛾周氏啮小蜂毒液对寄主美国白蛾两种血细胞包囊作用和吞噬作用的影响。【结果】美国白蛾颗粒细胞的包囊指数强于浆血细胞。白蛾周氏啮小蜂毒液对颗粒细胞和浆血细胞的包囊指数均有明显的抑制作用,毒液浓度越大,抑制作用越强,两种血细胞的包囊作用均呈先增长后降低的趋势。在所有浓度毒液处理下,颗粒细胞的包囊指数在12 h时最强。未经小蜂毒液处理的浆血细胞包囊指数在15 h时达到最强,但经浓度为0.01~0.03 VRE/μL的毒液处理后浆血细胞的包囊指数在12 h时达到最强,而经浓度为0.04~0.10 VRE/μL的毒液处理后包囊指数在9 h时最强。白蛾周氏啮小蜂毒液对美国白蛾蛹颗粒细胞的吞噬作用强于浆血细胞。毒液对两种血细胞的吞噬能力均有明显的抑制作用,但毒液处理对浆血细胞的吞噬作用影响较小。【结论】白蛾周氏啮小蜂毒液可以抑制美国白蛾蛹颗粒细胞和浆血细胞的包囊作用和吞噬作用,且随着毒液浓度的增加,两种血细胞的免疫作用显著下降。     

周氏啮小蜂非典型气味受体基因的cDNA序列及进化分析

【目的】对重大林业害虫美国白蛾Hyphantria cunea(Drury)的重要寄生性天敌周氏啮小蜂Chouioia cunea Yang(膜翅目:姬小蜂科)一个Or83b直同源基因的cDNA全长序列CcOr1进行多角度的进化分析,为研究周氏啮小蜂嗅觉分子机制奠定基础。【方法】通过转录组测序的方法鉴定CcOr1基因全长,并对该基因进行序列及进化分析。【结果】该基因长度为1 428 bp,编码475个氨基酸;蛋白二级结构预测具脊椎动物G蛋白典型的七跨膜结构域。【结论】该基因较为保守,主要接受负选择压力。作为寄主广泛的寄生蜂,周氏啮小蜂与寄主单一的类群相比,其同义突变率较低,密码子偏好性较高。     

Mass rearing and augmentative releases of the native parasitoid Chouioia cunea for biological control of the introduced fall webworm Hyphantria cunea in China

The fall webworm, Hyphantria cunea Drury, is an economically important introduced pest in China. A native pupal endoparasitoid, Chouioia cunea Yang (Hymenoptera: Eulophidae, Tetrastichinae) causes considerable mortality of H. cunea pupae in some areas of China. Alternative hosts of C. cunea were tested to find potential substitute hosts for mass rearing the parasitoid. Antheraea pernyi Guerin-Meneville (Lepidoptera: Saturniidae) was chosen as a surrogate host as a single pupa produced an average of 6552 wasps and a maximum of 11,256 per pupa. A mass-rearing technique and successful release program of this parasitoid is described. Parasitism by C. cunea in the areas where it was released averaged 67.74% and was usually over 80%. Other native parasitoids, such as Coccygomimus disparis (Viereck), C. parnasae (Viereck) (Hymenoptera: Ichneumonidae) and Exorita japonica Townes (Diptera: Tachnidae), caused the total average parasitism to exceed 90% on average, reaching a maximum of 96.28% in the release areas. These successful releases indicate that an introduced pest species can also be controlled by the mass-rearing and release of native parasitoids in the country of introduction.     

营养物质和水分对韭菜迟眼蕈蚊成虫繁殖和寿命的影响

【目的】研究韭菜迟眼蕈蚊Bradysia odoriphaga成虫对营养物质的选择补充习性,找出对其产卵繁殖起重要作用的关键因子,为其有效防控提供依据。【方法】通过室内成虫单独配对试验,研究不同营养物质(花粉、蜂蜜、维生素、糖、醋、酒)和水分对韭菜迟眼蕈蚊成虫繁殖和寿命的影响。【结果】随着花粉、蜂蜜和维生素浓度的增加,成虫的繁殖力下降;糖醋酒液单一成分和混合液对成虫繁殖和寿命的影响与水分相近;补充供试营养物质和水分对韭菜迟眼蕈蚊的繁殖力均明显高于干燥对照。补充水分的成虫繁殖力最高,产卵率超过70%,单雌平均产卵量达70粒,雌雄成虫寿命最长均超过3 d。土壤湿度对产卵有显著影响,土壤相对湿度为40%和50%时,成虫的落卵率超过25%,落卵量大于480粒,显著高于其它湿度(P<0.05)。【结论】水分是影响韭菜迟眼蕈蚊成虫繁殖和寿命的关键因子,供试的营养物质对韭菜迟眼蕈蚊繁殖没有促进作用,因此在韭菜迟眼蕈蚊成虫盛发期可以通过控制田间土壤湿度来影响韭菜迟眼蕈蚊繁殖。     

不同食物对烟粉虱三种寄生蜂取食寄生等行为的影响

【目的】就不同食物对烟粉虱Bemisia tabaci(Gennadius)寄生蜂取食、寄生等行为的影响进行分析,以阐明食物或营养来源对蚜小蜂取食、寄生等生物学行为的影响机制。【方法】以古桥桨角蚜小蜂Eretmocerus furuhashii、丽蚜小蜂Encarsia formosa和双斑恩蚜小蜂Encarsia bimaculata为研究对象,分别以蜂蜜水、2~3龄烟粉虱若虫及清水进行饲喂,观察和研究分析3种食物来源对3种蚜小蜂取食与寄生等行为的影响。【结果】3种蚜小蜂均以取食15%蜂蜜水的蚜小蜂的取食、搜索与寄生等行为频次最高、分配时间最长,而取食清水的蚜小蜂取食、搜索与寄生等行为频次最低、分配时间最短;取食不同食物后,3种蚜小蜂各种行为的变化趋势一致。【结论】不同食物或营养来源对蚜小蜂的取食及寄生等行为有着显著的影响,进而可以影响到蚜小蜂对烟粉虱的生物防治效率。     

Effect of zinc nutrition on sucrose biosynthesis in maize

Zinc deficiency caused an accumulation of ¹⁴C into malic acid, sugar phosphates, sugar nucleotides, glucose, fructose, phosphoenolpyruvate, glycine and alanine, whereas the ¹⁴C labelling in sucrose decreased. The activity of sucrose synthetase (EC 2.4.1.13) was unaffected up to the 15th day and thereafter it declined. Severe Zn deficiency reduced the biosynthesis of total protein and sucrose synthetase by 50 and 20%, respectively.     

Effect of nutrition on the LH response to calf removal and GnRH

Ten primiparous crossbred cows were assigned to two dietary groups at calving. One group received 120% and the other group received 80% of the National Research Council (NRC) recommended allowance of dietary energy for primiparous cows. At 60 days postpartum, calves were removed from their dams. Blood samples were collected from the cows at 15-min intervals for 8 hr beginning at the time of calf removal and again 24 hr, 48 hr and 72 hr after calf removal. At 72 hr after calf removal, all cows were given 200 ug GnRH intravenously. At calf removal, serum LH concentrations were higher (P<0.01) for cows on 120% (0.9 +/- 0.03 ng/ml) compared to cows on 80% (0.5 +/- 0.03 ng/ml) of recommendations. Serum LH concentrations increased (1.6 +/- 0.1 ng/ml, P<0.01) by 24 hr in cows on the highenergy diet. In contrast, a similar increase was not observed in cows on the low-energy diet until 48 hr after calf removal (1.4 +/- 0.2 ng/ml, P<0.01). These contrasting patterns in serum LH concentrations resulted in a diet by time interaction (P<0.01). Serum LH concentrations increased in both dietary energy groups following GnRH injection, but the response was greater (P<0.01) in cows on the low-energy diet compared to the cows fed the high-energy diet. These results indicate that inadequate dietary energy delays the LH response to calf removal and increases the LH response to exogenous GnRH.     

重大外来入侵害虫——美国白蛾生物防治技术研究

美国白蛾Hyphantria cunea(Drury)是重大外来入侵害虫,对我国林业和园林绿化造成重大危害,尤其是近年来美国白蛾传入北京,严重威胁着我国“绿色奥运”的顺利举办和首都园林绿化及生态环境安全。根据美国白蛾多发生在居民区、乡镇和城市的特点,作者开展对环境安全的生物防治技术研究。汲取前苏联等国家从美国白蛾的原产地引进天敌进行生物防治没有成功的经验和教训,立足调查和寻找我国本土天敌。经过21年的努力,发现和筛选出白蛾周氏啮小蜂Chouioia cunea Yang(新属新种)这种寄生美国白蛾的特优天敌;研究这种小蜂的行为学、生态学、生物学、解剖学及人工大量繁殖、放蜂防治技术;筛选出人工大量繁殖小蜂的替代寄主,解决小蜂的大量繁殖的瓶颈问题;通过放蜂防治试验,取得良好的防治效果。同时,筛选出防治美国白蛾的高毒力HcNPV病毒毒株,研制出美国白蛾人工饲料,利用人工饲料大量饲养美国白蛾幼虫,然后接种扩增病毒,成功解决HcNPV病毒大量扩增生产和常年生产的技术难题,做到病毒的规模化生产和常年生产,保证大面积生产防治的需要和病毒质量。研究出利用天敌昆虫和病原微生物综合应用的防治模式:在美国白蛾幼虫期喷洒HcNPV病毒,蛹期释放白蛾周氏啮小蜂,达到既控制当代美国白蛾的危害,又有效抑制其下代的种群数量,取得长期的持续控制美国白蛾的防治效果。上海市、大连市、烟台市和青岛市利用该项技术已经完全控制美国白蛾,特别是上海市利用本项生物防治技术防治后,已经连续6年没有再发现美国白蛾。由于该项生物防治技术利用的是自然界原有的控制害虫的生态因子,因而对环境安全,做到既消灭害虫,又保证生态环境和人畜的安全,也保护生物多样性。该项研究成果也为北京采用“绿色”防治技术有效控制美国白蛾提供重要的科技支撑和技术保障。     

Effect of nutrition on growth and virulence of the entomopathogenic fungus Beauveria bassiana

Three isolates of the entomopathogen Beauveria bassiana along with one strain of Metarhizium anisopliae were cultured on seven media with different carbon/nitrogen (C/N) ratios. The effect of nutrition on virulence of the isolates was evaluated via measurement of colony growth, spore yield, germination speed, conidial C/N ratio and Pr1 (a serine protease) activity. 'Osmotic stress' medium produced the lowest colony growth with low numbers of conidia in all isolates. However, these conidia showed a high germination rate and virulence. However, conidial Pr1 activity was low in some isolates. In most but not in all cases conidia from 1% yeast extract, 2% peptone and low (10 : 1) C/N medium had higher Pr1 activity compared with conidia from other media. However, in some instances we could not conclude that there was a relationship among germination rate, conidial Pr1 activity and virulence. C/N ratio of conidia was statistically different among various media and fungal isolates. Conidia with lower C/N ratio generally produced lower LT(50) (lowest median lethal time) values (more virulent). Insect-passaged conidia from different media had lower C/N ratio compared with similar conidia from artificial cultures. Therefore, they should be more virulent than in vitro produced conidia. As germination rate, conidial Pr1 activity and C/N ratio are independent of host, it seems that host-related determinants such as insect cuticle and physiology and environmental conditions may influence host susceptibility and therefore fungal isolate virulence towards host insects.     

Effect of potassium nutrition of rice on rhizosphere redox status

With four soils differing in K supplying power and with four rice cultivars (Oryza sativa L.) differing in K uptake kinetic parameters, the relationship between K fertilizer application and soil redox status in rhizosphere and; the distribution of ferrous iron and other toxic substances on the root surface and in the rhizosphere; and the effect of K supply on uptake of reduced iron by rice plants have been studied.The results show that K application on K-deficient soils reduced the content of active reducing substances and ferrous iron in the soil, raised the soil redox potential in the rhizosphere, increased the Eh value of rice roots and lowered the content of iron in the rice plants. These effects of K varied with different rice cultivars. When no K fertilizer was applied, active reducing substances and ferrous iron in rhizosphere soils were decreased more by the rice cultivars absorbing K strongly (e.g. Shanyou 64) than by cultivars absorbing K weakly (e.g. Zhongguo 91). Therefore, the diminution of the toxic substances by K application in the weakly K-absorbing cultivars was more significant.The observation of a rhizobox separated by a nylon screen showed that appreciably more iron oxides, compared with the control, were deposited at or adjacent to the root surfaces of the rice plant supplied with K fertilizer, fully demonstrating the relationship between K nutrition and the total oxidizing power of rice plants. According to the distribution of active reducing substances and ferrous iron, the oxidizing range of the rice root extended in K application treatment a few centimeters away from the root plane. K application to rice affected the soil redox status in rhizosphere in many ways. The main effect was an increase of the oxidizing power of the rice root. As a result, the value of soil Eh was increased, the contents of active reducing substances and ferrous iron were lowered, as well as the number of oxygen consuming microorganisms.