夹竹桃皂甙对福寿螺的毒杀效果及其对水稻幼苗的影响

室内条件下研究了夹竹桃皂甙对福寿螺的杀灭效果及其对水稻幼苗的影响。结果表明,福寿螺的致死率随着皂甙浓度的增加和处理时间的延长而显著上升,其中50mg/L 浓度皂甙处理48h的致死率达100%,与0.156g/L氯硝柳胺水溶液的灭螺效果相当。皂甙对福寿螺的杀灭效果与福寿螺的大小有关。皂甙对壳高h＜10 mm的福寿螺的杀螺效果最好, 而对20≤h<30 mm的杀螺效果最差。水稻幼苗的存活率随着皂甙浓度的增加而显著增加,且低浓度皂甙处理24h也具有较高的值。此外,采用水培方法研究了皂甙对水稻幼苗鲜重的影响。高浓度(≥40 mg/L)的皂甙处理7d显著地抑制了水稻幼苗根部和地上部分的鲜重,其中对根部的影响大于地上部分。当处理时间延长到14d时,皂甙对水稻幼苗鲜重的抑制作用逐渐减弱,仅50mg/L 皂甙处理对根部鲜重有显著地抑制。而在氯硝柳胺水溶液处理下,水稻幼苗停止生长。综合结果表明夹竹桃皂甙是一种环境友好且能高效防治福寿螺的灭螺剂。     

除草剂胁迫对空心莲子草叶甲种群的影响及应对策略

为探讨除草剂胁迫对空心莲子草叶甲种群的影响,寻求合理施用除草剂,有效保护叶甲,提高其控草效果的应对策略,利用叶片残留法测试了在草甘膦、农达、水花生净与35%苄·丁可湿粉等除草剂胁迫下空心莲子草叶甲卵孵化率,幼虫、成虫存活率以及蛹羽化率。结果表明被测试的4种除草剂均不同程度地降低空心莲子草叶甲卵孵化率,幼虫与成虫存活率及蛹羽化率,特别是水花生净28.0、35.0 g/L两种浓度处理,35%苄·丁可湿粉4.0 g/L浓度处理显著降低卵孵化率;农达13.4 g/L浓度处理显著降低1龄前期幼虫存活率,农达40.1 g/L与 53.4 g/L两种浓度处理显著降低2龄前期幼虫存活率;水花生净28.0 g/L浓度处理显著降低3龄前期幼虫存活率;草甘膦97.6 g/L与 122.0 g/L浓度处理、农达26.7 g/L与66.8 g/L浓度处理、35%苄·丁可湿粉2.0 g/L浓度处理显著降低后期蛹的羽化率;35%苄·丁可湿粉1.0 g/L浓度处理显著降低羽化后3 d雌虫的存活率。除草剂除直接杀死空心莲子草叶甲,降低其种群数量外,使叶甲失去食物与避护所,快速地引起叶甲种群崩溃。建议合理施药,并在施药时,随机留若干小斑块不施药,保护叶甲,可望提高其田间控草效果。     

皂荚提取物的杀螺活性

[目的]研究皂荚生物农药活性,开发利用皂荚资源,发展环境友好的绿色植物源农药。[方法]采用室内生测和田间试验研究皂荚壳乙醇提取物的杀螺活性。[结果]皂荚提取物对福寿螺有显著的毒杀活性,对幼螺和成螺72 h的LC₅₀分别为40.56、109.83 mg·L^-1。田间试验表明,皂荚提取物对福寿螺有较好的防效,施用40 g·m^-2的皂荚提取物处理7 d后卵块减少率为100.00%（成螺失去产卵的能力）,防效为（99.12±1.26）%。[结论]皂荚提取物对福寿螺较好的生物防治效果,是一种潜在的生物杀螺剂。     

雌雄福寿螺耐寒能力的差异

为了解福寿螺耐寒性是否存在性别差异,对华南地区的雌雄福寿螺进行了过冷却研究及低温胁迫下存活率调查,并对入冬前后其体内耐寒性生理生化物质进行测定.结果表明: 雌雄螺的过冷却点均值分别为-6.83 ℃和-6.26 ℃,雌螺过冷却点显著低于雄螺;进入冬季后,福寿螺自由水含量明显下降,结合水、甘油、脂肪、糖原含量均出现明显上升,除甘油外,其余各耐寒性生理指标均存在极显著的性别差异;未经过冷驯化的雌雄福寿螺经过10 ℃处理7 d后存活率均在90%以上,继续在5 ℃下培养7 d后存活率出现明显的性别差异和壳高差异.野外调查发现在福寿螺自然种群中雌螺多于雄螺.表明福寿螺雌螺的耐寒能力强于雄螺.这对进一步探索福寿螺的生态适应性、自然性比及其耐寒性机制有一定的参考价值.     

雌雄福寿螺耐寒能力的差异

为了解福寿螺耐寒性是否存在性别差异,对华南地区的雌雄福寿螺进行了过冷却研究及低温胁迫下存活率调查,并对入冬前后其体内耐寒性生理生化物质进行测定.结果表明:雌雄螺的过冷却点均值分别为-6.83℃和-6.26℃,雌螺过冷却点显著低于雄螺;进入冬季后,福寿螺自由水含量明显下降,结合水、甘油、脂肪、糖原含量均出现明显上升,除甘油外,其余各耐寒性生理指标均存在极显著的性别差异;未经过冷驯化的雌雄福寿螺经过10℃处理7d后存活率均在90%以上,继续在5℃下培养7 d后存活率出现明显的性别差异和壳高差异.野外调查发现在福寿螺自然种群中雌螺多于雄螺.表明福寿螺雌螺的耐寒能力强于雄螺.这对进一步探索福寿螺的生态适应性、自然性比及其耐寒性机制有一定的参考价值.     

干燥方式对尾巨桉树皮提取物总三萜含量及其杀螺活性的影响

为探讨干燥方式对尾巨桉树皮提取物总三萜含量的影响和其杀螺作用机理,实验以尾巨桉树皮为研究对象,比较鼓风干燥和冷冻干燥两种干燥方式对其树皮乙醇提取物总三萜含量及杀螺活性的影响。结果表明,当乙醇体积分数为60%时,鼓风干燥和冷冻干燥的尾巨桉树皮乙醇提取物中总三萜含量最高,分别为10.81%和13.90%。两种干燥方式得到的尾巨桉树皮乙醇提取物对福寿螺均有较强的毒杀活性,在50mg/L的浓度下处理72h,福寿螺的死亡率分别到达93.1%和100.00%;同时尾巨桉树皮提取物还能显著抑制福寿螺离水上爬,抑制效果优于同浓度下的对照药剂茶皂素(P<0.05)。与空白对照组相比,经鼓风干燥的尾巨桉树皮提取物处理后,福寿螺头足和肝脏的蛋白质含量变化不大(P>0.05);经过冷冻干燥的尾巨桉树皮提取物处理后,福寿螺头足中蛋白质含量显著降低(P<0.05),而肝脏中蛋白质含量无明显变化(P>0.05)。鼓风干燥和冷冻干燥的尾巨桉树皮提取物均可有效抑制福寿螺头足部中LDH、AKP和AST/GOT的活性(P<0.05)以及肝脏中ALT/GPT和AST/GOT的活性(P<0.05)。冷冻干燥的尾巨桉树皮提取物中三萜含量以及杀螺活性均高于鼓风干燥的树皮提取物。     

常用萃取剂提取福寿螺卵中类胡萝卜素的初步研究

【背景】福寿螺为世界性恶性入侵水生动物,也是我国公布的第一批外来人侵物种之一。福寿螺大量啃食为害水稻、茭白、白莲等重要农作物,对我国南方各省农业生产造成了巨大威胁,因此,预防和控制福寿螺灾害显得尤为重要。合理利用福寿螺能有效控制福寿螺的数量和危害,是生物防治的一个重要部分。福寿螺卵中含丰富的类胡萝卜素,充分利用螺卵中的类胡萝卜素能拓展福寿螺的利用途径和方法。【方法】为探寻福寿螺卵中类胡萝卜素的提取方法,本研究采用甲醇、无水乙醇、丙酮、乙酸乙酯、二氯甲烷、石油醚等6种常用萃取剂提取福寿螺卵中类胡萝卜素,用紫外可见光分光光度计测定其含量。【结果】结果显示,不同萃取剂中类胡萝卜素的提取量不同,醇类为较适合的提取液（甲醇〉无水乙醇〉丙酮）。【结论与意义】本研究对福寿螺卵中类胡萝卜素的提取方法进行了探索和研究,找出了合适的提取液,为拓展福寿螺的利用途径,以及福寿螺的综合防治提供了参考。     

乙草胺降解菌系的分离鉴定及其生长降解特性研究

乙草胺是一种主要用于防除禾本科杂草的酰胺类除草剂,苄嘧磺隆是一种用于防除阔叶杂草的磺酰脲类除草剂,乙·苄复配制剂由于扩大了除草谱而被广泛应用于稻田中,但大量使用也带来了环境隐患。本研究从长期施用乙·苄复配制剂的稻田土壤中筛选到1个可耐受600 mg/L苄嘧磺隆的乙草胺降解复合菌系,从中分离到4个细菌菌株66C-1、66C-2、66C-3、66C-4,利用形态、生理生化及分子生物学方法,分别鉴定为粘质沙雷式菌(Serratia marcescens)、Psedomona shunanensis、恶臭假单胞菌(Pseudomonas putida)和硝化还原假单胞菌(Pseudomonas nitroreducens)。这4个菌株等比例混合构成的降解菌系HXDW可以利用乙草胺为唯一碳源、但不能利用丁草胺、异丙甲草胺和吡氟酰草胺等其他酰胺类除草剂。降解菌系HXDW在乙草胺初始浓度为100 mg/L的无机盐培养基中,0.1%~2.0%Na Cl浓度范围内生长良好,最适培养温度为25℃。提取降解菌系不同部位粗酶液对50 mg/L乙草胺进行处理,胞外提取液的24 h乙草胺降解率为44.95%,表明降解菌系HXDW对乙草胺的降解作用主要存在于胞外。     

福寿螺的形态构造

本文对引进我国的一种大型淡水养殖螺类——福寿螺(大瓶螺)的外部形态、内部构造进行了描述,其中内部构造部分为国内首次系统报道,并附有形态解剖图。在解剖中我们发现,福寿螺有一些特殊的结构:(1)栉状鳃着生于外套腔的右侧,这与所属的中腹足目及瓶螺科的特征(鳃位于外套腔的左侧)明显不同。(2)胃的内壁有许多列隆起,其上有磨状齿,用以磨碎食物;胃腔凹面的底部有一呈倒U型的胃楯。(3)大动脉的基部膨大成副心脏,这在腹足纲动物中很少见。(4)雄性生殖器有前列腺囊,囊内的前列腺呈细丝状,长可达70毫米。     

厌氧丁草胺降解菌BAD-20的分离鉴定及降解特性研究

【目的】分离并鉴定能够降解除草剂丁草胺的厌氧微生物菌株,研究其厌氧降解丁草胺的特性和代谢途径,为深入研究丁草胺厌氧降解机制提供依据。【方法】以丁草胺为碳源作为选择压力从水稻田土壤中富集驯化丁草胺降解菌,利用16S rRNA基因系统发育分析结合菌株培养特征对降解菌株进行初步鉴定,利用液相色谱-时间飞行质谱(LC-TOF-MS)检测菌株降解丁草胺的代谢产物。【结果】筛选到一株降解丁草胺的厌氧细菌,命名为BAD-20,初步鉴定为嗜蛋白质菌属(Proteiniphilum),菌株BAD-20降解丁草胺的最适条件为温度30–35℃、pH 7.5–8.0和0–0.5%NaCl,在有氧条件下该菌不能降解丁草胺。最适条件下,菌株BAD-20在10d降解90%的20mg/L丁草胺。菌株BAD-20还能降解甲草胺、乙草胺、丙草胺,降解效率从高到低依次为甲草胺乙草胺丙草胺丁草胺,对这些氯乙酰胺除草剂的降解动力学符合一级动力学方程。鉴定到2个丁草胺降解代谢产物,分别是N-(2,6-二乙基苯基)-N-(丁氧甲基)乙酰胺(DEPBMA)和N-(2,6-二乙基苯基)乙酰胺(DEPA),表明菌株BAD-20降解丁草胺的起始步骤为脱氯,随后脱去N-丁氧甲基。【结论】本研究富集分离到一株降解丁草胺的厌氧细菌嗜蛋白质菌属(Proteiniphilum) BAD-20,为深入研究丁草胺厌氧降解机制及研发含丁草胺废水厌氧生物处理技术提供依据。     

in-silico analysis suggests alterations in the function of XisA protein as a possible mechanism of butachlor toxicity in the nitrogen fixing cyanobacterium Anabaena sp. PCC 7120

Butachlor, a commonly used herbicide adversely affects the nitrogen fixing capability of Anabaena, an acclaimed nitrogen fixer in the Indian paddy fields. The nitrogen fixation in Anabaena is triggered by the excision of nifD element by xisA gene leading to rearrangement of nifD forming nifHDK operon in the heterocyst of Anabaena sp. PCC7120. Functional elucidation adjudged through in-silico analysis revealed that xisA belongs to integrase family of tyrosine recombinase. The predicted functional partners with XisA protein that have shown cooccurence with this protein in a network are mainly hypothetical proteins with unknown functions except psaK1 whose exact function in photosystem I is not yet known. The focus of this study was to find out the relation between XisA and butachlor using in-silico approaches. The XisA protein was modeled and its active sites were identified. Docking studies revealed that butachlor binds at the active site of XisA protein hampering its excision ability vis-à-vis nif genes in Anabaena sp. PCC7120. This study reveals that butachlor is not directly involved in hampering the nitrogen fixing ability of Anabaena sp. PCC7120 but by arresting the excision ability of XisA protein necessary for the functioning of nif gene and nitrogen fixation.     

Construction and analysis of an intergeneric fusant able to degrade bensulfuron-methyl and butachlor

Rhodococcus sp. BX2 degrades bensulfuron-methyl but not butachlor, and Acinetobacter sp. LYC-1 degrades butachlor but not bensulfuron-methyl. Functional strains were constructed through protoplast fusion of Rhodococcus sp. BX2 and Acinetobacter sp. LYC-1 to generate fusants with an improved ability to simultaneously degrade bensulfuron-methyl and butachlor. Initial identification and stability tests of the fusants were performed. Three fusants with eighth transfer on plates containing two antibiotics and two herbicides were obtained. F1 also grew well in an inorganic salt solution containing bensulfuron-methyl and butachlor. F1 was characterized by its parents’ morphological and physio-biochemical features. F1 not only had bands in common with BX2 and LYC-1, but also had its own specific bands analyzed by Random Amplified Polymorphic DNA. The genetic similarity indices between F1 and BX2 and F1 and LYC-1 were 0.507 and 0.470, respectively. The percentages bensulfuron-methyl and butachlor degradation by F1 in an inorganic salt solution supplemented with 100 mg/L bensulfuron-methyl and 100 mg/L butachlor were 65.35 and 62.41 %, respectively, and the percentages in soil contaminated with 10 mg/kg bensulfuron-methyl and 10 mg/kg butachlor with an inoculum size of 5 % at 34 °C and at a pH of 7.5 after 35 days were 63.74 and 61.53 %, respectively. It was demonstrated that F1 could simultaneously degrade bensulfuron-methyl and butachlor.     

福寿螺对稻田水生植物群落实验的影响

福寿螺是在我国南方地区危害水稻生产的一种外来入侵软体动物,已被世界自然保护联盟入侵生物专家组列为100种危害最为严重的外来入侵物种之一。在水域生态系统中福寿螺主要以摄食各种水生植物而生存。以水稻、大薸、水葫芦、粉绿狐尾藻、水花生和鸭舌草构成的稻田水生植物群落为对象,研究了低密度（4 株/m²）、中密度（8 株/m²）和高密度（12 株/m²）福寿螺处理对稻田水生植物群落的影响,同时结合福寿螺的食物选择性实验分析其相关行为特性。结果表明：1）低密度、中密度和高密度福寿螺处理1周内水稻密度均急剧下降,高密度福寿螺造成水稻密度下降超过70%,2周后福寿螺危害水稻秧苗的程度减轻;2）各密度处理福寿螺均抑制了水花生和鸭舌草扩散,第8周时高密度福寿螺处理中水花生和鸭舌草的密度仅为对照的30%和25%。同时,低密度福寿螺处理中的水花生和鸭舌草的密度显著高于中密度和高密度处理。不同密度福寿螺处理对水葫芦、大薸和粉绿狐尾藻影响不明显;3）水稻的日均密度增长率为负值,低密度、中密度和高密度福寿螺处理间存在明显差异,且均显著低于对照。水花生和鸭舌草的日均密度增长率虽为正值,但显著低于对照,水葫芦、大薸和粉绿狐尾藻的日均密度增长率和对照无差异。福寿螺日均密度增长迅速,各密度处理无显著差异。福寿螺对水稻、水花生、鸭舌草的选择性显著高于水葫芦、粉绿狐尾藻和大薸;4） 福寿螺引入8周后稻田水生植物群落物种多样性Simpson指数、Shannon指数和均匀度指数均显著降低。Simpson指数的大小顺序为：对照 > 低密度 > 中密度、高密度,Shannon指数和均匀度指数的大小顺序为：对照 > 低密度 > 中密度>高密度。说明福寿螺对供试水生植物有明显的选择性,其入侵会严重危害稻田水生植物群落的物种多样性。     

除草剂苄嘧磺隆对粗梗水蕨配子体生长和性器官分化的影响

 稻田施用的除草剂可能会污染邻近水域,对非靶标水生植物产生影响。国家二级重点保护野生植物粗梗水蕨(Ceratopteris pteridoides)主要分布于稻田及其周围水域,其有性繁殖敏感阶段与除草剂施用时间一致,配子体易暴露在除草剂污染中。为了评价除草剂污染对水生态环境特别是对非靶标国家重点保护野生水生植物的影响,保护生物多样性和水生生态环境,该文研究了除草剂苄嘧磺隆(Bensulfuron-methyl)对粗梗水蕨配子体生长和性器官分化的影响。结果表明,苄嘧磺隆对粗梗水蕨孢子萌发没有影响,但对其配子体生长有抑制作用。苄嘧磺隆浓度和粗梗水蕨配子体面积呈明显的剂量效应关系,苄嘧磺隆对粗梗水蕨配子体生长抑制的EC₅₀(半数效应浓度)为0.086μg&;#8226;L^-1,低于已报道的环境背景浓度。随着苄嘧磺隆浓度升高,粗梗水蕨的雌雄同体配子体逐渐减少,且配子体上颈卵器形成时间推迟。在苄嘧磺隆为10μg&;#8226;L^-1时,粗梗水蕨没有雌雄同体配子体形成。以上研究结果表明,低浓度苄嘧磺隆可抑制粗梗水蕨配子体的生长并影响其性器官的分化,使粗梗水蕨有性繁殖能力减弱。     

Screen of Bacillus thuringiensis toxins for transgenic rice to control Sesamia inferens and Chilo suppressalis

Transgenic rice to control stem borer damage is under development in China. To assess the potential of Bacillus thuringiensis (Bt) transgenes in stem borer control, the toxicity of five Bt protoxins (Cry1Aa, Cry1Ab, Cry1Ac, Cry1Ba and Cry1Ca) against two rice stem borers, Sesamia inferens (pink stem borer) and Chilo suppressalis (striped stem borer), was evaluated in the laboratory by feeding neonate larvae on artificial diets containing Bt protoxins. The results indicated that Cry1Ca exhibited the highest level of toxicity to both stem borers, with an LC₅₀ of 0.24 and 0.30 μg/g for C. suppressalis and S. inferens, respectively. However, S. inferens was 4-fold lower in susceptibility to Cry1Aa, and 6- and 47-fold less susceptible to Cry1Ab and Cry1Ba, respectively, compared to C. suppressalis. To evaluate interactions among Bt protoxins in stem borer larvae, toxicity assays were performed with mixtures of Cry1Aa/Cry1Ab, Cry1Aa/Cry1Ca, Cry1Ac/Cry1Ca, Cry1Ac/Cry1Ba, Cry1Ab/Cry1Ac, Cry1Ab/Cry1Ba, and Cry1Ab/Cry1Ca at 1:1 (w/w) ratios. All protoxin mixtures demonstrated significant synergistic toxicity activity against C. suppressalis, with values of 1.6- to 11-fold higher toxicity than the theoretical additive effect. Surprisingly, all but one of the Bt protoxin mixtures were antagonistic in toxicity to S. inferens. In mortality-time response experiments, S. inferens demonstrated increased tolerance to Cry1Ab and Cry1Ac compared to C. suppressalis when treated with low or high protoxin concentrations. The data indicate the utility of Cry1Ca protoxin and a Cry1Ac/Cry1Ca mixture to control both stem borer populations.     

水浮莲对水稻竞争效应、产量与土壤养分的影响

为了解入侵植物水浮莲在稻田生产中的扩散规律和对水稻农业性状的影响,通过田间水稻田实验,按照de Wit 取代试验方法和添加系列设计方法研究水浮莲与水稻(云稻2 号)混种对植株形态和生物量影响、种间竞争效应、水稻产量以及对土壤养分的影响。结果表明,混种条件下水浮莲母株株高、分蘖数、生物量和开花株数均受到水稻的明显抑制,受到的抑制率显著高于其对水稻的抑制率。水浮莲的种间竞争大于种内竞争(RY 小于1.0)而水稻的种内竞争大于种间竞争(RY 大于1.0),水浮莲混种比例大于和等于1:1 时(RYT大于1.0)与水稻不存在竞争作用,而小于1:1时(RYT 小于1.0)其存在着竞争作用,水浮莲对水稻的竞争力(CB 小于0)小于水稻。混种条件下水稻有效穗数和产量有明显提高,增产比例为3.54%-13.38%。生长过程中水浮莲对土壤钾、磷元素消耗大于水稻,而有机质和氮元素消耗小于水稻;混种条件下水稻明显降低水浮莲对土壤养分消耗,且二者在土壤养分上没有竞争关系。所有这些表明,入侵稻田的水浮莲与水稻生长过程中其形态、生物量等方面都处于劣势,而且一定的水浮莲密度有利于抑制水浮莲对土壤养分的消耗和促进水稻生长繁殖及其产量的提高,因此为满足饲料利用和环境净化,建议在正常耕作稻田中可对水浮莲进行适当的应用。     

Reprint of: Saprolegnia strains isolated from river insects and amphipods are broad spectrum pathogens

Saprolegnia species are destructive pathogens to many aquatic organisms and are found in most parts of the world. Reports based on phylogenetic analysis suggest that Saprolegnia strains isolated from aquatic animals such as crustaceans and frogs are close to Saprolegnia strains isolated from infected fish or fish eggs and vice versa. However, it has often been assumed that host specificity occurs for each individual isolate or strain. Here we demonstrate that Saprolegnia spp. can have multiple hosts and are thus capable of infecting different aquatic organisms. Saprolegnia delica, Saprolegnia hypogyna, and 2 strains of Saprolegnia diclina were isolated from aquatic insects and amphipods while S. delica, Saprolegnia ferax, Pythium pachycaule, and a Pythium sp. were isolated from the water of a medium to fast flowing river. The ITS region of the rRNA gene was sequenced for all isolates. In challenge experiments, all four isolates from insects were found to be highly pathogenic to eggs of Atlantic salmon (Salmo salar) and embryos of the African clawed frog (Xenopus laevis). We found that Saprolegnia spp. isolated from salmon eggs were also able to successfully establish infection in nymphs of stonefly (Perla bipunctata) and embryos of X. laevis). These results suggest that Saprolegnia spp. are capable of infecting multiple hosts, which may give them an advantage during seasonal variation in their natural environments.     

Cladochytriales—a new order in Chytridiomycota

Recently, molecular and ultrastructural analyses have resulted in revised phylogenetic hypotheses in the phylum Chytridiomycota. The order Chytridiales, once considered monophyletic, has been subdivided into several new orders. However, the most recent analyses indicate that the emended Chytridiales is also polyphyletic. One monophyletic lineage in Chytridiales includes Cladochytrium, Nowakowskiella, and five other genera. Many of the chytrids in this clade have often been observed growing on decaying plant tissue and other cellulosic substrates from aquatic habitats and moist soils. In this study we analysed combined nu-rRNA gene sequences (partial SSU and LSU) of 30 isolates from North American aquatic and soil samples. Based on molecular monophyly and zoospore ultrastructure, we designate this clade as a new order, Cladochytriales, which includes four families: Cladochytriaceae, Nowakowskiellaceae, Septochytriaceae fam. nov., and Endochytriaceae.