甜菜夜蛾核型多角体病毒中国分离株的分离鉴定及毒力测定

本文对分离自中国的甜菜夜蛾病毒进行提纯、鉴定.生物测定的结果表明其对二龄、三龄甜菜夜蛾的LC50分别为6.6× 104,2.6×105PIB/mL.     

甜菜夜蛾核型多角体病毒中国分离株的分离鉴定及毒力测定

本文对分离自中国的甜菜夜蛾病毒进行提纯、鉴定。生物测定的结果表明其对二龄、三龄甜菜夜蛾的LC50 分别为 6 .6× 10 4,2 .6× 10 5PIB/mL .     

温度对甜菜夜蛾核型多角体病毒流行的影响

在恒温条件下 ,研究了甜菜夜蛾 3龄初幼虫感染核型多角体病毒后的病死速率、病死时间分布与温度关系。结果表明 ,在2 9℃以下 ,随温度的升高 ,病死率增加 ,幼虫病死速率加快 ,病死持续时间缩短 ;该病毒的热抑制温度在 2 7℃左右。改进的Schoolfield模型、Stinner模型可很好地描述幼虫病死速率与温度关系。甜菜夜蛾种群饲毒后的每日病死率可用时间 -剂量 -死亡率模型较好地拟合 ,模型模拟值与实测值有较好的吻合 (Hosmer- L em oshow统计量检验不显著 ) ,方程中各项系数经 t检验达极显著水平 ;不同温度下的幼虫累计病死时间分布可用 Weibull模型、Gompertz模型及 L ogistic模型拟合 ,模型经 F检验显著 ,模型中各系数经 t检验均达到或接近显著水平。用剩余平方和 Q比较各模型的拟合程度 ,以 L ogistic模型拟合最好 ,Gompertz模型次之 ,Weibull模型拟合效果稍差。上述模型可以用来模拟分析不同温度下的幼虫病死时间分布和幼虫病死速率     

不同营养条件下甜菜夜蛾幼虫感染核型多角体病毒的敏感性及取食趋性

为探讨不同营养条件下甜菜夜蛾Spodoptera exigua对甜菜夜蛾核型多角体病毒(Spodoptera exigua multiple nucleopolyhedrovirus,SeMNPV)的敏感性及取食行为响应,本研究通过改变甜菜夜蛾人工饲料配方建立高蛋白低糖(简称高蛋白)、中蛋白中糖(简称中蛋白)、低蛋白高...     

斜纹夜蛾核型多角体病毒分子生物学研究进展

斜纹夜蛾核型多角体病毒(Spodoptera lituramulticapsid nucleopolyhedrovirus,SpltMNPV)属NPV科A亚群。近几年有关该病毒的序列测定、基因结构、功能和表达调控等系统的分子生物学研究工作进展迅速,特别是对一些重要基因的结构分析,有助于筛选毒力较强的杀虫毒株,并为这一病毒杀虫剂的改良和发展以及组建昆虫杆状病毒表达载体奠定基础。综述了与SpltMNPV相关的分子生物学领域的研究进展。     

红棕灰夜蛾核型多角体病毒的研究

红棕灰夜蛾 Sarcopolia illoba(Butler)是鳞翅目夜蛾科害虫。分布于我国东北、华北、华东、华中及苏联、日本、朝鲜、印度等地。幼虫多食性,为害烟草、甘蓝、马铃薯、甜莱、大豆、豌豆、菜豆等作物。1984年在哈尔滨大豆田中,从该虫幼虫自然死虫体内分离出一株核型多角体病毒。我们对该病毒的形态学进行了观察。这种病毒在国内外尚未见报道。并对12种夜蛾科害虫:迹幽夜蛾(Discestra stigmosa)、甘蓝夜蛾(Mamestra brassicae)、黄地老虎(Scotia segetum)、小地老虎(S.ipsilon)、银纹夜蛾     

苜蓿丫纹夜蛾核型多角体病毒（AcMNPV）大立方形多角体突变株的分子…

利用空斑技术,从既能形成多角体又含ＴＫ酶基因的苜蓿丫纹夜蛾重组核型多角体病毒ＡｃＭＮＰＶ－ＴＫ中,纯化了一株形成大立方形多角体的病毒突变ＡｃＭＮＰＶ－ＴＤｍ１５１３。用Ｅｏｏｒｌ、ＲｓｔⅠ、ＢｇｌⅡ及ＫｐｎⅡ等限制性内切酶对它做了酶切分析,并克隆了多角体蛋白全基因及其侧翼部分的片段。对所克隆的部分了理测定结果,发现在多角体蛋白基因读码框内只出现了一个碱基的突变,导致了第２５位的氨基酸密码子由ＧＧＴ     

甜菜夜蛾核多角体病毒泛素基因的克隆及原核表达

甜菜夜蛾核多角体病毒（Spotoptera exigua multi-nucleopolyhedrovirus,SeMNPV)泛素基因ubiquitin被克隆和序列分析,该基因编码区全长243bp,编码80个氨基酸残基,预计蛋白质分子量为9.4kDa。将这一ubiquitin基因克隆到原核表达载体pET-28a上,转化至BL21（DE3）中,用IPTG进行诱导表达,对表达的条件进行优化,用异源的泛素单克隆抗体检测目的蛋白,Western blot 实验证明所表达的蛋白是泛素蛋白。同时,我们制备了特异性的抗体,为以后的研究工作做了基础,通过计算机软件Gendoc对不同来源的泛素进行分析,结果显示,病毒中的泛素与真核细胞中的泛素相比较,泛素的氨基酸序列有较大的变化,杆状病毒的深入素基因在分子进化上可能有比较独特的途径。     

甜菜夜蛾核型多角体病毒对拟澳洲赤眼蜂的影响：种群参数比较和PCR检测

用生命表方法和PCR检测研究了甜菜夜蛾核型多角体病毒（Spodopteraexiguamulticapsid nucleopolyhedrovirus, SeMNPV）对拟澳洲赤眼蜂Trichogramma confusum Viggiani的影响。结果表明,在25±1℃下,取食含SeMNPV蜂蜜和纯蜂蜜（对照）的两组赤眼蜂雌蜂的寿命、繁殖力、羽化率、产雌率均无显著差异;内禀增长率（r_m）分别为0.3524 和0.3418,周限增长率（λ）为1.42和1.41,非常接近;净繁殖率（RO）分别为 124.34和142.57,平均世代历期为13.6和14.5天, 种群倍增时间分别为1.96和2.03天。对5头取食含SeMNPV蜂蜜的雌蜂的子代的PCR 检测结果也均为阴性。因此,结论认为SeMNPV对拟澳洲赤眼蜂成蜂并无明显影响,体内携带病毒的亲代蜂并不能将病毒传给子代。     

寄主植物对甜菜夜蛾三龄幼虫抗寒力的影响

寄主植物是影响昆虫抗寒性的主要因子之一。研究了不同温度下甜菜夜蛾Spodoptera exigua(Hübner)3龄幼虫取食小白菜、甘蓝、葱和菠菜后,对过冷却能力和体内冷冻保护剂的影响。结果表明,寄主植物对甜菜夜蛾3龄幼虫的过冷却能力有显著性影响,其中以取食甘蓝的幼虫过冷却点最低。温度和寄主植物对其过冷却点、结冰点和虫体含水量有明显的交互作用。寄主植物对其体内的海藻糖含量有显著性影响,而对甘油和糖原含量没有显著性影响。温度和寄主植物仅对海藻糖含量有显著的交互作用。随着温度的升高,取食不同寄主的幼虫体内海藻糖和糖原含量的变化趋势完全相反,认为海藻糖是由糖原转化而来。研究结果提示冬季合适的寄主植物有利于甜菜夜蛾低龄幼虫越冬。     

寄主植物对甜菜夜蛾的发育和繁殖及体内酯酶活性的影响

在恒温(2 8±1 )℃条件下考察了葱、苋菜、豇豆、蕹菜、菜心5种寄主植物对甜菜夜蛾SpodopteraexiguaH櫣ｂｎｅｒ的营养效应及体内羧酸酯酶、乙?铛ッ富钚缘挠跋?,结果表明,不同寄主植物对甜菜夜蛾幼虫和蛹的发育历期、存活率、蛹重、雌成虫产卵量等有显著影响,其中对雌成虫产卵量的影响最大。雌成虫产卵量最高为取食菜心的处理,其次是取食葱的处理。寄主植物对甜菜夜蛾体内羧酸酯酶、乙酰胆碱酯酶活性也有显著影响,但这些酶的活性与发育历期、存活率、产卵量无明显相关性。在进行预测预报、田间防治时应充分考虑寄主植物对甜菜夜蛾发育历期、产卵量、解毒酶活性的影响。     

甜菜夜蛾在四种寄主植物上的生命表参数比较研究

在控制条件下对甜菜夜蛾Spodoptera exigua(Hübner)在白菜、大葱、甘蓝和豇豆上的生命表参数进行了比较研究。结果表明,甜菜夜蛾幼虫在4种寄主植物上均为5个龄期。幼虫和蛹在大葱上的发育历期最长,在白菜上最短。甘蓝叶片饲养的甜菜夜蛾生殖力最高,单雌产卵量为1015.8粒,豇豆叶片上饲养的生殖力最低,为496.1粒。甜菜夜蛾在甘蓝上的内禀增长率和净增殖率最高,分别为0.237和287.82,在大葱上最低,分别为0.172和173.90。在大葱上甜菜夜蛾幼虫存活率较低,在其他3种寄主植物上较高。甜菜夜蛾的特定年龄生殖率在甘蓝叶片上最高,第22天单雌产卵量高达453.6粒。研究结果表明,在选取的4种植物中,甘蓝是甜菜夜蛾的最适寄主。     

异色瓢虫规模化生产技术及瓢虫工厂的建立

异色瓢虫Harmonia axyridis(Pallas)是一种重要的捕食性天敌,工厂化人工饲养异色瓢虫是利用其进行蚜虫等小型害虫生物防治的有效手段。本文报道了在河南省济源白云实业有限公司建立的基于甜菜夜蛾Spodoptera exigua(Hübner)低龄幼虫的工厂化生产的技术流程。利用这一技术,可以大量、稳定地生产异色瓢虫各虫态个体,基本实现了异色瓢虫的工厂化生产。文中还探讨了异色瓢虫产业化进程中存在的问题,期望瓢虫天敌产业化和商业化能够有所突破。     

Physiological response of Colorado potato beetle and beet armyworm larvae to depletion of wound-inducible proteinase inhibitors in transgenic potato plants

Larvae of Colorado potato beetle (CPB), Leptinotarsa decemlineata, and beet armyworm (BAW), Spodoptera exigua, reared on potato plants in which wound-induced accumulation of proteinase inhibitors (PIs) was largely reduced through antisense-mediated depletion of a specific lipoxygenase (LOX H3) had significantly larger weight gains than those fed on non-transformed plants. The midgut endoproteolytic activities of CPB larvae fed on non-transformed potato were significantly higher than those from larvae fed on LOX-H3-deficient plants. However, none of these proteolytic activities was inhibited by potato leaf extracts, regardless of the plant that they were fed on. Taken together, these data suggest that CPB, a leaf-feeding specialist of solanaceous plants, is largely adapted to the inducible PIs of potato, though the metabolic cost associated with the hyperproduction of digestive proteases may account for the 14-31% lower weight gain of larvae fed on non-transformed plants. The effect of LOX-H3 depletion on insect performance was more evident with larvae of the polyphagous BAW (52-63% higher weight gain and 73% higher fecundity when reared on LOX-H3-deficient plants). The poorer larval performance of BAW on non-transformed plants may be due to the susceptibility to inhibition by potato leaf tissues of most BAW digestive proteases. Indeed, BAW larvae fed on non-transformed potato showed a significant reduction in most endoproteolytic activities compared to larvae fed on LOX-H3-deficient plants, suggesting a that these insects deal poorly with induced plant defences in potato.     

寄主植物对松墨天牛幼虫体内酶系活性的影响

室内采用在马尾松、黑松、火炬松、湿地松和雪松皮下接入松墨天牛Monochamus alternatus Hope幼虫的方法,探明5种寄主植物对松墨天牛幼虫的存活、钻蛀取食及纤维素酶、酯酶、羧酸酯酶活性的影响。结果表明,5种寄主植物对松墨天牛幼虫存活和钻蛀取食有显著影响,接种于马尾松的天牛幼虫存活率100%;接种于雪松的松墨天牛幼虫存活率仅55.56%。松墨天牛取食不同寄主后,其体内纤维素酶、酯酶和羧酸酯酶活性存在显著差异。其中以取食火炬松的Cx酶活性最高,为(4.5754±0.1975)μmol·g-1(FW)·min-1,C1酶和β-l,4-葡萄糖苷酶活性以取食马尾松的松墨天牛幼虫消化道中最高,分别为(2.8199±0.0510)μmol·g-1(FW)·min-1与(3.0793±0.1733)μmol·g-1(FW)·min-1;而以取食黑松和马尾松的酯酶、羧酸酯酶活性最高,分别为1372.74mmoD/min·μg和744.82mmoD/min·μg。     

甜菜夜蛾飞行肌中与能量代谢有关的酶活性

通过生化方法测定了参与飞行能源物质代谢的5种酶活力的变化,即3-磷酸甘油醛脱氢酶(GAPDH)、3-磷酸甘油脱氢酶(GDH)、乳酸脱氢酶(L DH)、柠檬酸合酶(CS)和3-羟酰辅酶A脱氢酶(HOAD)。结果表明,1、3、5日龄处女蛾的GAPDH和CS酶活力显著高于7日龄蛾;但HOAD的活性恰恰与之相反,其酶活力随日龄的增加逐渐增强。GDH初羽化时活性较低,但到3日龄后增加到最大值,以后随日龄的增加活性逐渐下降,L DH的活性一直比较低,且不同日龄的酶活力差异不显著。交配状况和幼虫期饥饿处理对成虫的代谢酶活力没有显著影响。在第7代选育家系中,强飞家系的GAPDH酶活力显著高于短飞和不飞;而在第8代中强飞家系的上述5种酶的活性均高于短飞家系,而第10代中仅有CS的活性在强飞和短飞家系中存在显著差异,其他均差异不显著。这说明甜菜夜蛾在初始阶段主要利用糖类作为能源物质,而后期飞行又可通过脂类代谢提供能量,且不同日龄的处女蛾和选择家系的飞行能力与代谢酶活性呈现一定的正相关关系。     

不同寄主植物继代饲养后草地贪夜蛾幼虫生理酶活性差异

为了明确继代饲喂不同寄主植物对草地贪夜蛾幼虫体内生理酶活性的影响,在室内分别用3种寄主植物叶片饲养草地贪夜蛾多代,测定其F3代5龄幼虫体内保护酶（SOD,POD,CAT）、消化酶（脂肪酶、胃蛋白酶和α-淀粉酶）和解毒酶（GSTs,CarEs,CYP450s）活性差异,并分析不同寄主植物次生代谢物含量与幼虫体内酶活及其生长发育的相关性。结果表明,取食马铃薯叶片的F3代幼虫总发育历期最长,取食小麦的最短,两者间差异显著;小麦叶片中的单宁、总酚、类黄酮和生物碱含量均显著低于玉米和马铃薯;草地贪夜蛾幼虫体内保护酶活性均为取食小麦叶片的最低,取食马铃薯叶片的最高,其中取食马铃薯叶片的草地贪夜蛾体内POD和SOD酶活性达取食小麦的1.34和1.26倍;以3种寄主为食的F3代幼虫脂肪酶和α-淀粉酶活性变化不大,但持续饲喂马铃薯叶片3个世代可导致草地贪夜蛾幼虫体内胃蛋白酶活性显著升高,达10.502 U/mg. prot,为持续饲喂玉米叶片幼虫（1.508 U/mg. prot）的6.96倍以上。连续饲喂3种寄主叶片后的幼虫GSTs和CarEs活性无显著性差异,但取食玉米叶片的幼虫体内细胞色素P450s（CYP450s）活性显著高于取食小麦和马铃薯的幼虫。可见,连续多代取食次生物质含量不同的寄主植物会使草地贪夜蛾体内部分酶活性发生变化,并对幼虫的生长发育产生影响。     

In vivo development of the entomogeneous hyphomycetePaecilomyces farinosus in hostSpodoptera exigua (beet armyworm) larvae

The in vivo development of the entomogenous hyphomycetePaecilomyces farinosus inSpodoptera exigua (beet armyworm) larvae was examined using light and electron microscopic techniques. Blastospores injected into larval hemocoels (500 blastospores/larva) were immediately ingested by phagocytic hemocytes, and no fungal cells were detected in the hemolymph until 36 h post-injection. As indicated by immunocytochemical methods, the in vivo-produced blastosopres, in contrast to in vitro blastospores, lacked a galacto-mannan surface layer required for opsonization by aS. exigua humoral lectin. Therefore, these in vivo cells were not recognized by phagocytic granulocytes and were freely-circulating in the hemolymph. Hyphae differentiating from the blastospores were recognized by the hemocytes and induced formation of multicellular hemocytic nodules. By 72 h post-injection, mycelia were observed emerging from the nodules and by 96 h, larvae had become mummified due to extensive proliferation of the fungus throughout host tissues. Neither phagocytosis of the initially injected in vitro-produced blastospores nor nodule formation around hyphal cells later in the infection process was effective in stopping fungal growth. The in vivo development ofP. farinosus was similar to that of another hyphomycete,Beauveria bassiana except that in the latter case, extensive nodule formation was inhibited by the production of fungal metabolites.     

Prolonged root hypoxia effects on enzymes involved in nitrogen assimilation pathway in tomato plants

In order to investigate the effects of root hypoxia (1–2% oxygen) on the nitrogen (N) metabolism of tomato plants (Solanum lycopersicum L. cv. Micro-Tom), a range of N compounds and N-assimilating enzymes were performed on roots and leaves of plants submitted to root hypoxia at the second leaf stage for three weeks. Obtained results showed that root hypoxia led to a significant decrease in dry weight (DW) production and nitrate content in roots and leaves. Conversely, shoot to root DW ratio and nitrite content were significantly increased. Contrary to that in leaves, glutamine synthetase activity was significantly enhanced in roots. The activities of nitrate and nitrite reductase were enhanced in roots as well as leaves. The higher increase in the NH₄⁺ content and in the protease activities in roots and leaves of hypoxically treated plants coincide with a greater decrease in soluble protein contents. Taken together, these results suggest that root hypoxia leaded to higher protein degradation. The hypoxia-induced increase in the aminating glutamate dehydrogenase activity may be considered as an alternative N assimilation pathway involved in detoxifying the NH₄⁺, accumulated under hypoxic conditions. With respect to hypoxic stress, the distinct sensitivity of the enzymes involved in N assimilation is discussed.Key words: tomato, hypoxia, nitrogen, glutamine synthetase, protease, glutamate dehydrogenase