Sub-lethal effects of Beauveria bassiana(Balsamo) on field populations of the potato tuberworm Phthorimaea operculella Zeller in China

The potato tuberworm Phthorimaea operculella Zeller, is one of the most important potato pests worldwide including China. Several reports indicate that P. operculella could be controlled biologically by the use of beneficial fungus such as Beauveria bassiana(Bals.-Criv) Vuill. However, limited information is available under growing conditions in China. Thus, this study evaluated the sub-lethal effects of B. bassiana on the offspring of P. operculella by the age-stage, two-sex life table. First instar larva of P. operculella were treated with 1×107 conidia m L–1 of the fungus, and several biological parameters were evaluated. The fecundity, duration of the egg stage, all larval stages, pre-adult stage, and total pre-oviposition period, were significantly shorter than the control treatment. Offspring of treated parents, presented a net reproductive rate and mean generation time of 17.43 per day and 24.98 days, respectively, compared to 65.79 per day and 26.51 days for the untreated ones. This study provides basic information to help understanding the potential long-term effects of entomopathogenic fungi on P. operculella.

Lethal and Sub-Lethal Effects of Transgenic Rice Containing cry1Ac and CpTI Genes on the Pink Stem Borer, Sesamia inferens(Walker)

Lethal and sub-lethal effects of transgenic rice containing cry1Ac and CpTI genes on the pink stem borer, Sesamia inferens, were studied to collect information for ecological risk assessment on insect-resistance of transgenic rice. In vitro insect-feeding bioassays were conducted to evaluate the lethal and sub-lethal effects of transgenic rice lines （II YouKF6 and KF6） containing cry1Ac＋CpTI genes on S. inferens at four different growth stages, viz., seedling, tillering and elongation, booting, and milk and maturing. Transgenic rice at seedling stage showed significantly high lethal effect on S. inferens with the shortest lethal duration for 50 and 100% individuals and the highest corrected mortalities after feeding on transgenic lines at this stage for 3 and 6 d. Followed by tillering and elongation stage, 50 and 100% S. inferens were dead after feeding on transgenic lines at this stage for 4 and 10 d, respectively. Moreover, corrected mortalities for 6 d feeding on transgenic lines at this stage were significantly higher than that at booting, and milk and maturing stages. Lethal effect of KF6 on S. inferens decreased significantly at booting stage. Lethal duration for 50% S. inferens significantly extended and its corrected mortalities for 6 d feeding also declined remarkably. However, lethal effect of II YouKF6 on S. inferens did not decrease significantly at this stage. Transgenic rice at booting, and milk and maturing stages did not show significant lethal effect to S. inferens and it showed the longest lethal duration for 50% individuals and the lowest corrected mortalities for 3 and 6 d feeding. A few larvae of S. inferens could survive, pupate and emerge on these two transgenic lines at booting, and milk and maturing stages. Sub-lethal effect of two transgenic lines on S. inferens also differed significantly between different developmental stages. Continuously feeding on transgenic rice lines at seedling, and tillering and elongation stages delayed the development of larvae and pupae and decreased pupation rate, but no effect was observed on eclosion rate. Larval development was significantly inhibited after feeding on transgenic rice at booting stage, but no obvious effect was observed in pupal stage, pupation and eclosion rate. There were no significant differences for larval and pupal development, pupation, and eclosion rates between feeding on transgenic and control rice lines at milk and maturing stage. Larval and pupal weights significantly declined, but no influence was observed on fecundity when S. inferens infested on transgenic rice at any stage. These showed that there were significant differences in lethal and sub-lethal effects of transgenic rice on S. inferens among developmental stages, and the effects gradually decreased with the increase of growth stages of rice plant.

Specific Lethal Effects of Trace Neutral Red on Amoeba discoides and Trichodina

[ Objective] The aim of this study was to provide the reference for the artificial breeding of aquatic animals. [ Method] Amoeba discoides and Trichodina were treated by neutral red solution with different concentrations to observe the physiological activities of organelles in their death processes. Effects of neutral red solution on the growth of common aquatic animals such as Paramecium caudatum, Euglena viridis and Brachionus plicatilis were analyzed, and the specific lethal mechanism of trace neutral red to A. discoides and Trichodina was also preliminarily studied. [ Re- suit] The neutral red solution at a concentration of 0.5 mg/L damaged the physiological function of contractile vacuole in Trichodina and also had the specific lethal effect on Tdchodina, but it had no effect on the growth and reproduction of non-parasitic protozoa and B. p/icatilis. Neutral red so- lution with certain concentration led to disorder of the physiological functions of A. discoides, such as assimilation and rejection, which was a main factor that caused the death of A. discoides. [ Conclusion] With the advantages such as targeting, safety and easiness to be oxidized and decom- posed, neutral red is an ideal drug for treating the diseases caused by A. discoides and Trichodina, and its suitable concentration is 0. 5 mg/L.

Lethal effect of mononuclear cells derived from human umbilical cord blood differentiating into dendritic cells after in vitro induction of cytokines on neuroblastoma cells

BACKGROUND： Dendritic cell is the most major antigen presenting cell of organism. It is proved in recent studies that human umbilical cord blood mononuclear cells induced and cultured in vitro by recombinant human granulocyte-macrophage colony stimulating factor （rhG-MCSF） and recombinant human interleukin-4 （rhlL-4） can generate a great many dendritic cells and promote the lethal effect of T cells on human neuroblastoma, but it is unclear that whether the lethal effect is associated with the most proper concentration of dendritic cells. OBJEETIVE： To investigate the lethal effect of human umbilical cord blood mononuclear cells induced in vitro by cytokines differentiating into dendritic cells on human neuroblastoma, and its best concentration range. DESIGN ： Open experiment SEI-FING： Department of Pediatrics, the Medical School Hospital of Qingdao University MATERIALS ： The study was carried out in the Shandong Provincial Key Laboratory （Laboratory for the Department of Pediatrics of the Medical School Hospital of Qingdao University） during September 2005 to May 2006. Human umbilical cord blood samples were taken from the healthy newborn infants of full-term normal delivery during October to November 2005 in the Medical School Hospital of Qingdao University, and were voluntarily donated by the puerperas. Main instruments： type 3111 CO2 incubator （Forma Scientific, USA）, type 550 ELISA Reader （Bio-Rad, USA）. Main reagents： neuroblastoma cell line SK-N-SH （Shanghai Institute of Life Science, Chinese Academy of Sciences）, RPMI-1640 culture fluid and fetal bovine serum （Hyclone）, rhlL-4 （Promega, USA）, rhG-MCSF （Harbin Pharmaceutic Group Bioengineering Co.Ltd）, rat anti-human CDla monoclonal antibody and FITC-labeled rabbit anti-rat IgG （Xiehe Stem cell Gene Engineering Co.Ltd）. METHODS： ① Human umbilical cord blood mononuclear cells obtained with attachment methods differentiated into human umbilical cord blood dendritic cells, presenting typical morphology of dendritic cells after in vitro induction by rhG-MCSF and rhlL-4. ② Different concentrations of dendritic cells[ dendritic cells： neuroblastoma cells=20：1,50：1,100：1 （2×10^8 L^-1,5×10^8 L^-1,1×10^9 L^-1）], 1×10^9 L^-1 T cells and 1×10^7 L^-1 neuroblastoma cells were added in the experimental group. 1 ×10^9 L^-1 T cells and 1 ×10^7 L^-1 neuroblastoma cells were added in the control group. ③ Main surface marker CDla molecules of dendritic cells were detected with indirect immunofluorescence, and the percent rate of dendritic cells was counted with ultraviolet light and expressed as the expression rate of CD1a^＋ cells. ④Single effector cells and target cells were respectively set in the experimental group and control group to obtain the lethal effect. The lethal effect of dendritic cells on neuroblastoma cells was indirectly evaluated by detecting cellular survival with MTT assay. The lethal effect（%）= （1-A experimentat well-A effector cell /A target cell well）×100%.⑤The expenmental data were presented as Mean ±SD, and paired t test was used. MAIN OUTCOME MEASURES： ① Morphological characters of dendritic cells in the process of induction and differentiation. ②CD1a^＋ cellular expression rate. ③Lethal effect of dendntic cells on neuroblastoma cells. RESULTS： ①Morphological characters of dendritic cells in the process of induction and differentiation： On the 15^th day after human umbilical cord blood mononuclear cells were induced by rhG-MCSF and rhlL-4, typical morphology of dendritic cells could be seen under an inverted microscope. ②Expression rate of CD1a^＋ cells was （43.12±5.83）%. ③Lethal effect of dendritic cells on neuroblastoma cells： Lethal effect of dendritic cells stimulated T cells in each experimental group （ dendritic cells： neuroblastoma cells=100：1,50：1, 20：1 respectively） on neuroblastoma cells was significantly higher than that in control group[（31.00 ±4.41 ）%, （30.92±5.27）%,（33.57±5.35）%,（26.23±5.20）%, t=3.51,2.98,4.24, P〈 0.01 ）; But the lethal effect of dendntic cells on neuroblastoma was significantly lower when their ratio was 100：1 and 50：1 in comparison with 20：1 （t=2.01,2.36, P 〈 0.05）, and no significant difference in lethal effect existed between the ratio at 100：1 and 50：1 （t=0.06,P 〉 0.05）. CONCLUSION： Dendritic cells differentiated from human umbilical cord blood mononuclear cells after in vitro induction of cytokines can promote the lethal effect of T cells on neuroblastoma cells. The lethal effect is associated with the concentration of dendritic cells within some range.

Effect of Purified Paper Wasp Ropalidia marginata Venom Toxins on Different Biomolecules in Mice Serum

This study evaluated the effects of purified paper wasp Ropalidia marginata venoms on various biomolecules in the blood serum of albino mice. Changes in the concentration of some important macromolecules, i.e., proteins, free amino acids, uric acid, cholesterol, pyruvic acid, total lipids and glucose were noted down. These alterations were measured after intraperitoneal injection of 40% and 80% 24-hour LD50 purified Ropalidia marginata venom toxin. Serum total protein levels were found to decrease to 78% after 6 hrs, while serum free amino acid levels were significantly increased to 117% 6 hrs after venom injection compared to control. It was also found that serum uric acid levels increased to 138% after 8 hrs of venom injection compared to control. The increase in serum cholesterol i.e. (101% and 106%) and pyruvic acid increased significantly to a maximum value of 106% after 6 hrs of treatment at 40% LD50. Glycogen levels in the gastrocnemius muscle were found to decrease significantly (p-0.05) to 43% and 92% at LD50 after injection of purified Ropalidia marginata venom after 8 h and 80% at LD50 compared to control. Moreover, up to 71% and 81% were obtained at 10 hrs of treatment with the same dose. In the present study, the purified toxins significantly changed the levels of biomolecules in blood serum, indicating their wider effects on cellular physiology due to toxic effects and stress on the animal. These toxins can be good antigens and stimulate immune responses in experimental mice.

The acute lethality and endocrine effect of 1,2,3,7,8-PeCDD in juvenile goldfish (Carassius auratus) in vivo

1,2,3,7,8-PeCDD was administrated to juvenile goldfish （Carassius auratus） by peritoneal injections to explore the acute lethality and endocrine effects of 1,2,3,7,8-PeCDD in vivo. The value of acute median lethal dosage （LD50） of 1,2,3,7,8-PeCDD was determined in acute lethality tests. The endocrine effect of 1,2,3,7,8-PeCDD, whose exposed concentrations were determined based on the LD50 （1.84 mg/kg）, was studied by measuring the plasma vitellogenin （Vtg） content in juvenile male goldfish with enzyme-linked immunosorbent assays （ELISA）. Due to its significant induction of the plasma Vtg after one week＇s exposure in vivo in the 1/2 LD50 and LD30 groups, 1,2,3,7,8-PeCDD might be one of the important contributors to the estrogenic effect of PCDDs in the environment. The values of 1/2 LD50 and LD30 were within the range of the effective dosages of 1,2,3,7,8-PeCDD, indicating that there was a certain relationship between the estrogenic effective dosages and the LD50.

咖啡果小蠹的假死行为及短时高温对其致死作用

咖啡果小蠹[Hypothenemus hampei(Ferrari)]是2019年新入侵海南的一种严重威胁全世界咖啡生产的害虫,对海南甚至全国咖啡产业的健康发展构成重大威胁。为尽快了解咖啡果小蠹的习性,以及为今后该害虫的防控技术提供基础数据,本研究观察了咖啡果小蠹的假死行为和耐饥能力,并探讨短时高温对其致死效应。结果表明:24 h不饲喂任何食料对咖啡果小蠹成虫的假死行为无显著影响;45℃短时高温处理2 h显著提高了咖啡果小蠹成虫出现假死反应的比例和时长,假死反应出现比例达72.0%,假死时长为320.33 s,显著高于26℃下的假死时长(43.26 s)。咖啡果小蠹成虫具有较强的耐饥能力,在不提供任何食料的情况下,其最长存活时间可达204 h,平均存活时长达90.17 h。咖啡果小蠹成虫经短时高温处理后,处理温度与处理时长均显著影响其存活率,随着处理温度的升高及处理时间的延长,死亡率逐渐增加。45℃处理6 h后校正死亡率达94.92%;处理12 h的死亡率达100%;36℃和39℃处理6 h及以下时长对咖啡果小蠹的存活能力影响较小,36℃和39℃处理6 h,处理后24 h的校正死亡率分别为16.94%和23.73%。不同短时高温处理的半致死时间(Lt50)表现为随处理温度的升高而缩短,且随着观察时间的延长,Lt50随着缩短,其中45℃处理后24 h和48 h的Lt50最短,分别为3.11 h和2.70 h;36℃处理的Lt50最长,分别为17.85 h和11.42 h。不同短时高温处理6 h,处理后24 h和48 h的半致死温度(LT50)分别为39.53℃和38.66℃,LT50随观察时间的延长而降低。

定量磷酸化蛋白质组解析17β-雌二醇致死效应的细胞调控过程

17β-雌二醇(E2)是人体内一种重要的内分泌激素,在生理浓度下(0.2~1.0 nmol/L)对生殖系统、乳腺等靶器官的生长发育起着重要的调节作用。但很多研究表明,高剂量(μmol/L~mmol/L)的E2能够诱导肿瘤组织消退和细胞凋亡,其具体调控机制尚不明确。本工作聚焦于高剂量(μmol/L)的E2致死效应,首先分析了μmol/L水平的E2对HeLa细胞表型的影响,发现在1~10μmol/L下E2以浓度依赖的形式抑制HeLa细胞增殖,并诱导HeLa细胞发生死亡,其中,用5μmol/L E2处理2天后可使约74%的HeLa细胞增殖受到抑制,并引起约50%的HeLa细胞死亡。在此基础上,为了探究高剂量E2诱导细胞死亡的内在调控过程,将基于固相萃取(SPE)的固定化钛离子亲和色谱技术(Ti 4+-IMAC)与基于数据非依赖采集模式(DIA)的蛋白质组定量技术结合,用于筛选HeLa细胞内参与高剂量(μmol/L)E2致死效应调控过程的磷酸化位点。最终,在5μmol/L E2和二甲基亚砜(DMSO)处理的HeLa细胞中共鉴定到超过10000个磷酸化位点;t检验分析发现,在E2处理后,有924个磷酸化位点(对应599个蛋白质)的丰度发生了显著变化(显著性水平(p)<0.01,|log 2(倍数变化)|≥1),推测其可能参与调控E2致死效应过程。此外,有453个磷酸化位点(对应325个蛋白质)仅单独发生在E2或DMSO处理后的HeLa细胞样品中,表明这些磷酸化位点在E2处理后发生了磷酸化或去磷酸化,也可能参与E2致死效应的调控过程。分别对以上两种方式筛选的E2调控的磷酸化蛋白质进行富集分析,发现这些磷酸化蛋白质主要参与细胞分裂、核糖体/核质转运、信使核糖核酸(mRNA)加工/剪接及转录等过程,表明高剂量的E2可能通过调控核糖体及mRNA加工等过程影响蛋白质转录,进而诱导细胞发生死亡。此外,我们发现表皮生长因子受体(EGFR)和丝裂原活化蛋白激酶(MAPK)家族蛋白(包括MAPK1、MAPK4和MAPK14)上多个磷酸化位点的修饰水平在高剂量E2处理后发生了明显变化,表明EGFR和MAPK信号通路可能在雌激素诱导的细胞死亡中起着重要调控作用。本实验得到的磷酸化蛋白质组定量结果有助于进一步了解高剂量E2的内在调控过程,为后续解析高剂量E2的作用机制及疾病的治疗提供了参考。

Dose-effect correlation of chloride de-icing salt on Euonymus japonicus

In order to prevent severe pollution by de-icing salt on greenery along urban roads, a half lethal dose （LD_50）for a plant population was confirmed through stress simulation of chloride de-icing salt on Euonymus japonicus, with an ianalysis of physiological changes, statistics on mortality rate on plant populations and mathematic modeling during a 30- day subacute toxicity test. The results indicate that a significant positive correlation in the early stages and a significant negative correlation in the later stages were observed between the amount of chlorophyll a and b in plants and a cumulative dose of de-icing salt. The amounts of free proline in plants and the dose of de-icing salt were positively correlated Over the entire period. No significant correlation in the initial stage, but a significant negative correlation in later stages was observed between the soluble protein and the dose of de-icing salt. LDs0 of this chloride agent on E. japonicus is 5 kg.（L·m2）-1 over 30 days.

农药对熊蜂的生态毒理研究进展

熊蜂(Bombus spp.)因其独特形态特征及生物学特性成为农业生产中重要传粉昆虫,在维持自然及农业生态系统平衡发挥重要作用。近年来,熊蜂多样性呈减少趋势,农药不合理施用为重要因素之一。本文基于农药对熊蜂的毒理学研究现状,系统整理了熊蜂急性致死和慢性亚致死毒理研究方法,围绕农药对熊蜂的急性毒性,亚致死剂量农药对熊蜂精子活性及体温调节等生理发育,颜色分辨及学习记忆行为、肠道微生物结构及解毒因子等方面影响展开综述,对研究中存在的问题和未来研究方向进行了思考和展望,旨在为农药的科学使用、生物环境安全评估及熊蜂多样性保护提供参考。

反潜战斗部对耐压壳结构的毁伤威力评估方法与实验

针对反潜(鱼雷/深弹)战斗部多类型、水下爆炸多效应以及目标结构多样式等特点,研究归一化的战斗部毁伤威力表征与评估方法。基于最具代表性的艇体结构(耐压壳)毁伤,综合考虑大变形、撕裂破口和聚能穿孔等效应,提出一种通过综合毁伤威力指数量化表征并评估战斗部毁伤威力的方法,适用于各种典型反潜战斗部以及典型单层、双层壳体结构。设计模拟单层、双层壳体结构的毁伤效应靶标,进行爆破、聚能和聚爆3种模拟战斗部的海上试验,分别获得了3种战斗部对两种壳体结构的综合毁伤威力指数,并据此完成了量化对比与分析。研究结果表明:所提评估方法可量化给出同一战斗部对不同艇体结构,以及不同战斗部对同一艇体结构的毁伤威力差别;同等条件下,爆破和聚能战斗部分别对单层、双层壳体靶标具有相对更大的毁伤威力,聚爆战斗部对两种类型结构靶标伤威力均优于单纯的爆破或聚能战斗部;不同战斗部对双层壳体靶标较单层壳体靶标的毁伤威力均有所下降,其中爆破战斗部下降90%以上,聚爆战斗部下降近80%,而聚能战斗部下降只有不到20%。

6种活性化合物对旋毛虫各期虫体的体外杀伤作用

为了筛选有效活性化合物对旋毛虫不同发育时期虫体的体外杀伤作用,采用体外培养的方式测定三氯生等6种活性化合物对旋毛虫的体外杀伤效果,用倒置显微镜观察旋毛虫形态并计数。结果显示,三氯生对旋毛虫肌幼虫、成虫、新生幼虫均有明显杀伤作用(P<0.01),没食子酸辛酯对旋毛虫成虫和新生幼虫有明显杀伤作用(P<0.01),吡咯-2-羧酸、邻羟基苯乙酸、亚精胺盐酸盐、1,5-戊二胺仅对旋毛虫新生幼虫有杀伤作用(P<0.05)。亚精胺盐酸盐在高浓度下对肌幼虫有杀伤作用但效果不佳。结果表明,这6种活性化合物对旋毛虫各时期有虽有不同程度的杀伤作用,但只有三氯生对旋毛虫3个时期均具有明显的杀伤效果,该筛选结果为进一步开展体内试验研究提供了依据。

取食Cry3Aa蛋白后黄曲条跳甲成虫中肠组织的病理变化

为明确Cry3Aa蛋白对黄曲条跳甲成虫的致死效果及作用机理,采用浸叶法测定了黄曲条跳甲成虫取食Cry3Aa蛋白后的死亡情况,并利用透射电镜观察了Cry3Aa蛋白处理后成虫中肠组织的病理学变化。结果显示,取食浓度为2 mg/mL的Cry3Aa蛋白6 d和12 d后黄曲条跳甲成虫的死亡率分别为33.33%和77.78%;透射电镜观察发现,蛋白处理后成虫中肠发生了明显的病理变化,并且随着时间的延长,病理变化更加明显,主要表现为:柱状细胞变形、细胞间隙增大、微绒毛缩短并肿胀、线粒体变形、内质网肿胀。本研究表明黄曲条跳甲成虫中肠是Cry3Aa蛋白的重要作用部位,研究结果为揭示Cry3Aa蛋白对黄曲条跳甲成虫的作用机理奠定了基础。

多杀霉素协同绿豆含水量对绿豆象的致死效果研究

利用多杀霉素结合控制绿豆含水量,测试对绿豆象的致死效果,以期减少多杀霉素的使用量,延缓抗性的产生。控制绿豆含水量采用拌种法测试了多杀霉素对绿豆象成虫与卵的致死效果。结果表明,绿豆象卵与成虫的死亡率随多杀霉素浓度的增加和绿豆含水量的降低而升高;绿豆象卵对多杀霉素更敏感,利用多杀霉素对绿豆象成虫的亚致死LC_(10)浓度处理绿豆象的卵,其死亡率高达65%以上。建议利用多杀霉素控制绿豆象时,将绿豆含水量控制在9.5%~11.5%范围较佳。研究结果为绿豆的绿色仓储治理提供了参考。

生物化学农药 S- 烯虫酯防治储粮害虫研究进展

储粮害虫综合治理策略包括使用熏蒸剂和防护剂。当前,一些长期使用有机磷防护剂已在世界范围内逐步被淘汰,并被更安全的杀虫剂取代,如拟除虫菊酯和昆虫生长调节剂(IGR)。IGR包括保幼激素类似物(JHA)、蜕皮激素和几丁质合成抑制剂,因其昆虫特异性被认为是安全的。烯虫酯是储藏物害虫防治中使用最广泛的JHA。最初在储藏物中使用的烯虫酯配方中含有R-和S-形式的外消旋混合物,但现在只使用S-烯虫酯异构体。几十年来,烯虫酯受到了广泛的关注,主要集中研究其直接致死效应,但最近研究更多地关注其亚致死效应。文章全面综述国内外烯虫酯发展历程和现状,特别强调其对储藏物害虫的防治作用。

灰飞虱海藻糖酶基因的克隆及RNA干扰效应

RNA干扰(RNAi)不但可以用于研究基因的功能,还可以通过沉默靶标基因干扰特定的生命过程。因此,通过深入研究,发掘高效专一性靶基因和RNAi技术,有可能开辟针对性的害虫RNAi防控新途径。本研究通过灰飞虱Laodelphax striatellus转录组数据分析并结合RACE技术,克隆了灰飞虱两种海藻糖酶的全长基因,分别命名为LSTre-1和LSTre-2,其GenBank登录号分别为JQ027050和JQ027051。它们均具有海藻糖酶基因的典型特征,与已报道的其他昆虫的海藻糖酶基因具有很高的相似性,并表现出一定的虫种亲缘关系。其中LSTre-1为水溶性海藻糖酶基因,全长2042 bp,开放阅读框编码602个氨基酸,前端有25个氨基酸的信号肽,但无跨膜结构域;LSTre-2为膜结合型海藻糖酶基因,全长2619 bp,开放阅读框编码618个氨基酸,前端有26个氨基酸的信号肽,有2个疏水性跨膜结构域。利用喂食法研究2种海藻糖酶基因dsRNA对灰飞虱的致死效应,发现靶向水溶性酶基因的干扰效应略高于靶向膜结合型的,但两种海藻糖酶基因的dsRNA都可以显著抑制灰飞虱海藻糖酶基因的表达,降低其活力,还能显著抑制试虫的生长,大幅增加试虫死亡率。结果提示,通过适宜途径干扰海藻糖酶基因可以开发防治灰飞虱的新途径。

诱集植物香根草对二化螟幼虫致死的作用机制

【目的】香根草(Vetiveria zizanioides)能够有效诱集水稻害虫二化螟(Chilo suppressalis)雌成虫在其上产卵,但孵化出的幼虫在香根草上不能完成生活史。本研究旨在明确香根草对二化螟幼虫致死的作用机制,为开发以香根草为基础的水稻螟虫可持续控制新技术提供理论依据。【方法】通过有机溶剂萃取和硅胶柱层析方法提取香根草有毒活性物质,并利用饲料混毒法测定各提取组分对二化螟3龄幼虫的生物活性,同时利用生物化学方法测定水稻和香根草中营养物质含量以及二化螟3龄幼虫取食水稻和香根草后体内相关解毒酶和消化酶的活性变化。【结果】香根草石油醚萃取物处理二化螟3龄幼虫9、12、15 d后,死亡率分别为38.89%、57.41%、85.19%,显著高于乙酸乙酯萃取物、正丁醇萃取物等其他提取组分处理二化螟幼虫后的死亡率。进一步从香根草石油醚萃取物中分离获得组分1(Fr1)和组分5(Fr5),对二化螟3龄幼虫的具有较高毒性,以0.05 g·mL^(-1)浓度混入人工饲料饲喂3 d后死亡率分别为85.00%和67.67%,说明香根草中含有对二化螟幼虫具有致死作用的有毒活性物质。香根草中总蛋白、纤维素、总糖、氨基酸等营养物质含量分别为9.45μg、1.61%、1.45%、4.00%,均显著低于水稻中相应营养物质含量,其中水稻中蛋氨酸含量是香根草中的7.0倍。而香根草中单宁含量则显著高于水稻中的含量,是水稻中的1.31倍。此外,二化螟取食水稻和香根草3 d后,取食香根草的3龄幼虫体内蛋白酶、淀粉酶、海藻糖酶和蔗糖酶等消化酶的活性显著低于取食水稻幼虫体内相应消化酶的活性;6 d后,取食香根草的3龄幼虫体内细胞色素P450酶活性显著低于取食水稻的幼虫;9 d后,取食香根草的3龄幼虫体内羧酸酯酶CarE的活性显著低于取食水稻的幼虫。与取食水稻的3龄幼虫相比,取食香根草的幼虫体内谷胱甘肽硫转移酶GSTs的活性也有所降低,但是两者在统计学上没有显著差异。【结论】香根草作为二化螟的致死型诱集植物,对二化螟幼虫的致死作用主要表现在两方面:一是香根草中含有对二化螟幼虫具有致死作用的有毒活性物质,这些物质通过抑制幼虫体内解毒酶CarE和P450酶的活性,使幼虫逐渐丧失解毒代谢能力,最终死亡;二是香根草相对水稻营养物质匮乏,二化螟幼虫取食香根草后营养不均衡,从而影响体内消化酶活性,造成消化功能紊乱,影响其正常生理活动,最终死亡。

悬浮物对虾夷扇贝致死效应的研究

虾夷扇贝原产于太平洋西部和北部的俄罗斯、朝鲜及日本沿岸等地,适于在低温、高盐度海洋环境中生存、生长。为确定虾夷扇贝适宜的生存环境,研究了悬浮物对虾夷扇贝的致死作用。结果表明:(1)悬浮物对虾夷扇贝的急性致死效应(72h)不强;(2)高浓度悬浮物对虾夷扇贝具有很强的慢性致死作用。

转cry1Ac和CpTI双基因抗虫水稻对二化螟和大螟的致死效应及田间螟虫构成的影响

就转crylAc＋CpTI双基因抗虫水稻不同生育期对二化螟Chilo suppressalis和大螟Sesamia inferens的室内致死特性及田间螟虫的构成进行了研究。室内测定结果表明，不同生育期转基因水稻对二化螟、大螟都表现明显的致死效应，但水稻生长后期的致死效果降低。转基因水稻对大螟的致死效应显著弱于对二化螟的，其中，二化螟除在齐穗期和成熟期有少量幼虫（0．5％～6．4％）存活到第4天外，其余均在第4天死亡；大螟在两种转基因水稻上的存活率高于二化螟，且少量个体（〈1．6％）还能化蛹、羽化，但化蛹率和羽化率均明显低于在非转基因对照上的。早、晚两季水稻的田间调查结果表明，转基因水稻上两种螟虫虫口数量均显著低于相应的非转基因对照品种，转基因水稻上二化螟虫口减退率〉99％；大螟虫口减退率相对较低，早、晚稻上有所不同，其中早稻〉93％，晚稻仅44％～64％。转基因水稻上残存螟虫中，大螟所占比例明显上升，推测转基因水稻对两种螟虫致死效应差异可能是其主要原因。

转cry1Ac＋CpTI基因水稻对大螟的致死和亚致死效应

【目的】研究转cry1Ac+CpTI基因水稻对大螟的致死和亚致死效应,为转基因抗虫水稻的生态风险评价提供依据。【方法】采用活体取食生测的方法系统研究苗期、分蘖拔节期、孕穗期和灌浆成熟期4个生育期的转cry1Ac+CpTI基因水稻KF6和ⅡYouKF6对大螟的致死和亚致死效应。【结果】两转基因水稻在苗期对大螟的致死效应最为明显,其对50%和100%个体的致死作用时间最短,大螟在该生育期的转基因水稻上取食3d及6d后,校正死亡率最高;其次为分蘖拔节期,两转基因水稻在4.0d和10.0d内能分别对50%和100%的大螟个体产生致死作用,在该生育期的两转基因水稻上取食6d后,校正死亡率均明显高于孕穗期和灌浆成熟期;KF6在孕穗期对大螟的致死效应显著下降,其对50%大螟个体的致死作用时间显著延长,且取食6d后的校正死亡率也显著下降。但ⅡYouKF6对大螟的致死效应并没有明显下降;两转基因水稻在灌浆成熟期对大螟的致死效应显著下降,对50%大螟个体的致死作用时间显著长于其它生育期,取食3d和6d后的校正死亡率均显著低于其它生育期;部分大螟能在转基因水稻的孕穗期和灌浆成熟期完成个体发育。不同生育期的两转基因水稻对大螟的亚致死效应也存在显著差异,取食苗期和分蘖拔节期的转基因水稻能抑制大螟幼虫和蛹的发育,降低化蛹率,但对羽化率没有显著影响;取食孕穗期的转基因水稻能显著抑制大螟幼虫的发育,但对蛹历期、化蛹率和羽化率均无影响;灌浆成熟期的转基因水稻对幼虫和蛹的发育速率、化蛹率和羽化率均无明显影响。在任一生育期的转基因水稻上取食,均能降低大螟幼虫的体重和蛹重,但不影响成虫的生殖力。【结论】不同生育期的转基因水稻,对大螟的致死和亚致死效应存在显著差异,随生育期的增加,致死和亚致死效应逐渐减弱。