红树林沉积物Bt菌株的分离与cry基因型的鉴定（Isolation of Bacillus thuringiensis Strains from Mangrove Sediment Samples and Identification for cry Genes）

从收集自泉州洛江红树林保护区的400个沉积物土壤样品中,分离到18株苏云金芽孢杆菌(Bacillus thuringiensis,Bt).利用PCR-RFLP体系对Bt菌株杀虫晶体蛋白的基因型进行了鉴定,发现15株Bt含有cry1基因,4株含有cry2基因,有3株分别含有cry7、cry8、cry9基因.克隆了一个新型的cry2Ab基因,其编码蛋白与现有Cry2Ab型杀虫晶体蛋白的最高同源性为95%.     

多年连续种植转基因Bt汕优63稻田水体和土壤中Cry1Ab/c蛋白残留调查

以湖北武汉、随州、武穴和孝感以及山东德州5个多年转基因水稻种植区为试验地,在水稻生育期采集转cry1 Ab/c基因水稻Bt汕优63(Bt-SY63)和对照非转基因汕优63(SY63)稻田水体和土壤,使用酶联免疫吸附测定(ELISA)方法对稻田水体和土壤Cry1 Ab/c蛋白残留量进行动态监测.结果显示,与SY63稻田相比,连续种植2～4 a后,不同生育期Bt-SY63稻田水体Cry1Ab/c蛋白残留量大多与同一生育期SY63样品间差异未达显著水平(P＞0.05),最高残留量为0.373 ng· mL-1.与水体Cry1Ab/c蛋白残留情况相似,不同生育期Bt-SY63稻田土壤Cry 1Ab/c蛋白残留量大都低于试剂盒检测限(0.25 ng·g-1),仅随州苗期、德州拔节期和开花期样品有微量残留,鲜土残留量分别为0.261、0.540和0.361 ng·g-1,并分别与SY63样品间差异显著(P＜0.05).另外,多年连续种植Bt-SY63水稻后,2、3和4a种植年限的Bt-SY63稻田水体和土壤中Cry1Ab/c蛋白残留量之间大都没有显著差异(P＞0.05).认为连续种植2、3和4a的Bt-SY63水稻稻田水体和土壤仅存在微量Cry1Ab/c蛋白残留,不会造成Cry1Ab/c蛋白在稻田水体和土壤中的累积.     

基于环介导等温扩增法可视化检测玉米转基因成分

快速准确的转基因检测技术是严格监管转基因作物及其产品的重要支撑;不同转基因作物由于导入不同外源目的基因从而表现出不同性状,因此外源目的基因序列可作为基于核酸检测技术进行转基因筛选的标志物.基于环介导等温扩增(Loop-mediated isothermal amplification,LAMP)技术,以中性红染料作为比色试剂,建立玉米内源基因和玉米转基因成分的基因特异性的可视化快速检测方法.针对玉米内源基因IVR基因,抗虫外源bar、pat基因,和Bt176中的抗除草剂外源cry1Ab基因分别设计了LAMP引物.结果表明,在Real-time LAMP体系下,能够实现对玉米内源IVR基因、转基因玉米品系Bt176中的bar基因和转基因玉米品系TC1507和59122中的pat基因的检测.对转基因玉米品系Bt176中的cry1Ab基因设计的引物能特异性扩增Bt176,而对Bt11和MON810两种转基因玉米品系无效.通过优化Tris-HCl和中性红浓度,在5 mmol/L Tris-HCl和100μmol/L中性红浓度条件下建立了可视化检测体系,反应1 h即可分别实现对玉米内源基因IVR基因,抗虫外源bar、pat基因,和抗除草剂外源cry1Ab基因的检测.中性红可视化检测结果与Real-time LAMP荧光扩增曲线结果一致,体现了该可视化检测方法具有较高的准确性.本研究建立了一种对玉米转基因相关成分的可视化检测体系,该方法既快速又简便且尤其适用于现场检测.(图6表1参17)     

苏云金芽孢杆菌新基因cry1Ab17的克隆和生物信息学

利用高效克隆PCR产物的专用载体pMD18T,直接从苏云金芽孢杆菌WB9的PCR产物中克隆了cry1Ab17新基因.测序结果表明,该基因(GenBank登录号为AY646166)由3471个碱基组成,其编码的蛋白质含有1156个氨基酸残基,其中亲水性氨基酸占30.8%,疏水性氨基酸占45.2%,酸性氨基酸占12.9%,碱性氨基酸占11.1%.氨基酸序列的同源性分析结果表明,Cry1Ab17蛋白与已报道的Cry1Ab蛋白同源性为95.4%～99.7%,该蛋白的4个氨基酸残基———Pro170、Gly449、Gly796和Gly863与其它已报道Cry1Ab蛋白相应位置的氨基酸残基均不同.在核苷酸序列和氨基酸序列多重比较的基础上,应用PAUP4.0构建了Cry1A蛋白家族的系统发育树.SignalP分析结果显示,Cry1Ab17蛋白中不含信号肽序列.此外,对Cry1Ab17蛋白的二级结构和3个结构域也进行了预测和分析.图4表2参15     

转Bt-cry1Ac棉花花粉对意大利蜜蜂生长发育的影响

研究了取食转Bt-cry1Ac棉花花粉对意大利蜜蜂的影响,主要包括意大利蜜蜂的生长发育、体内酶活性的变化.结果发现,取食转Bt-cry1Ac基因棉花花粉的意大利蜜蜂与取食非转基因亲本棉花花粉的蜜蜂(CK)相比,4、5、6日龄幼虫体重差异不显著,幼虫及蛹的历期也没有明显差异.取食转Bt-cry1Ac基因棉花花粉的意大利蜜蜂6日龄幼虫体内的谷胱甘肽-S-转移酶、总蛋白酶活力与CK相比,没有显著差异.取食转Bt-cry1Ac基因棉花花粉的意大利蜜蜂幼虫体内的α-乙酸萘酯酶、乙酰胆碱酯酶活力极显著高于CK,强碱性、弱碱性类胰蛋白酶活力极显著低于CK,类胰凝乳蛋白酶活力显著低于CK.另外,采用酶联免疫(ELISA)方法在取食转Bt-cry1Ac基因棉花花粉的蜜蜂6日龄幼虫体内能够检测到Bt杀虫蛋白.表4参12     

mCry1Ac基因在抗虫转基因玉米Bt799中的表达特征

采用双抗夹心酶联免疫法(ELISA)定量检测转mCry1 Ac基因抗虫玉米Bt799在不同生育期不同植株部位的mCry1Ac蛋白含量。结果显示:mCry1 Ac基因在整个生育期玉米叶、茎、根和种子中均能表达,mCry1Ac蛋白含量为(0.82±0.10)~(15.83±1.77)μg·g-1;随着生育期和植株部位的不同,mCry1Ac蛋白含量呈现明显的时空动态变化,其中,叶、茎和根中mCry1Ac蛋白含量随转基因玉米生育期的推移均呈增加趋势,并均在完熟期达最高;在除苗期外的其他各生育期叶中mCry1Ac蛋白含量均显著高于其他植株部位,而在完熟期种子中mCry1Ac蛋白含量在各植株部位中最低,为(2.86±1.71)μg·g-1。     

苏云金芽胞杆菌YBT-1520中SigL及其增强子结合蛋白的结构与功能分析

为揭示SigL及其增强子结合蛋白(EBPs)在苏云金芽胞杆菌(Bt)中的调控功能,在全基因组测序的基础上,采用生物信息学方法,对YBT-1520菌株的SigL及其EBPs的结构和功能进行深入分析.结果表明,YBT-1520基因组中存在1个SigL和6个EBPs,而且EBPs在结构上具有丰富的多样性,包含了EBPs的所有可能的结构域组织类型.SigL所调控的基因涉及11个假定的COG代谢途径,其中包括能量代谢、氨基酸代谢、翻译与细胞周期等.根据EBPs在基因组的位置推测,YBT-1520的EBPs参与γ-氨基丁酸代谢途径、精氨酸代谢途径、支链脂肪酸降解途径、多糖分解代谢等代谢途径的调控.本研究将为揭示Bt杀虫晶体蛋白大量表达的调控机制提供新的思路.     

城市居民区圆柏花粉浓度的时空变化及其影响因素

通过对散粉期间居民区建筑不同位置、不同高度的圆柏花粉浓度进行了全天昼夜监测。结果表明居民楼群环境对圆柏花粉的时空分布有一定的影响,具体变化规律为:1)城市居民区内圆柏花粉浓度日变化为双峰单谷型,正午前后是首次高峰期,平均浓度为25粒/(h.cm2),凌晨左右为次高峰期,谷值出现在下午18:00时到人们晚上休息之前,平均浓度为2粒/(h.cm2)。2)总体上城市楼群边缘圆柏花粉浓度垂直分布梯度为1.5 m>13.5 m>7.5 m,楼群中心圆柏花粉浓度垂直变化梯度为13.5 m>7.5 m>1.5 m。但是不同时段内,由于主导因素不同,花粉浓度的垂直分布不尽一致。最后,还分析了气象要素对圆柏花粉浓度的影响。     

一株对鳞翅目害虫高毒力的苏云金芽孢杆菌菌株基因型鉴定及新基因cry2Ah3克隆

HYW-8是本实验室自行分离的一株对鳞翅目害虫具有高毒力的苏云金芽孢杆菌,PCR-RFLP鉴定含有cry1Ab、cry1E基因和一种新的cry2A基因,根据cry2A全长基因序列设计特异引物,获得全长基因.测序结果表明,该基因开放阅读框为1 896 bp,编码632个氨基酸,相对分子质量(Mr)是70.774×103,等电点pI=7.79,与已知的cry2Ah2基因编码的蛋白的同源性最高为98%.该基因序列已在GenBank中登记注册,登录号为GU073380,并被国际Bt杀虫晶体蛋白基因命名委员会正式命名为cry2Ah3.SDS-PAGE分析,HYW-8菌株表达130×103大小蛋白,未见70×103大小蛋白.     

磺胺类抗生素复合污染降解菌的筛选及其降毒作用

磺胺类抗生素是环境中检测出率较高的一类污染物.微生物降解是一种相对安全、高效且成本低的污染物去除技术,而关于磺胺类抗生素复合污染降解菌对该类抗生素的去除及其降毒能力方面的研究较少.因此,以3种磺胺类抗生素:磺胺吡啶(SP)、磺胺氯哒嗪(SCP)和磺胺二甲嘧啶(SM2)为碳源,从土壤中筛选3种磺胺类抗生素复合污染的降解菌,应用时间毒性微板分析法,测定3种抗生素及其复合污染物在降解前、后对指示生物蛋白核小球藻的毒性效应,并分析降解菌对3种抗生素及其复合物污染物的降毒能力.结果表明,以SP、SCP和SM2为碳源筛选出2株(S1和S2)降解菌,其中,S1菌株对3种抗生素的降解能力优于S2菌株,并初步鉴定S1降解菌为马氏棒杆菌属;S1降解菌的生长状况和降解能力最佳条件:3种抗生素的混合浓度为1 500 mg·L-1,pH=7.0,温度30℃,转速为150 r·min-1以及接种量为2.0％;S1菌株降解后的抗生素及其复合污染物对蛋白核小球藻的毒性明显低于降解前的毒性,降毒率在99％以上;S1降解菌对3种磺胺类抗生素的降解能力为SM2＞SP＞SCP.     

中国水稻土碳循环研究进展

文章首先分析了水稻土在碳循环研究中的地位和重要性，进而对我目水稻土碳循环的研究现状作了较为详尽的阐述，对其主要研究结论进行了深入的剖析。中国大而积的水稻土自1980年以来显示出有机碳库增加现象，说明水稻田对大气CO2可能产生汇效应。水耕熟化过程足有机碳的积累过程，水稻土的碳密度是早作土碳密度的2-3倍。水稻土的同碳能力与土壤的微团聚体的粒径有关。但对于水稻土中有机碳的分布和结合状态与农业管理措施、水稻土质量变化、农业生态环境变化的关系仍不清楚。因而建议就这一问题从土壤物理学、化学和生物学的相互作朋与土壤微团聚体中矿物质、有机质和微生物的相互结合关系的层面上进行多学科研究。同时提出了今后我困水稻土碳循环的重点研究方向和领域，即从整体和系统的角度来研究碳循环和平衡，从不同的时空区域来研究碳循环的过程和强度。     

Evaluating losses in ecosystem services in nature reserves in Sichuan,China

Nature reserves have developed rapidly over the decades in China and play a significant role in the function of ecosystem services. The function of ecosystem services in nature reserves, however, has tended to decline in recent years due to natural hazards and human activities. Based on land use, the variation of ecosystem services value (ESV) during 2000–2010 in national nature reserves in Sichuan was evaluated. We analyzed the comprehensive effects of natural disasters and human activities on the decline of ESVs. Total ESV in the national nature reserves in Sichuan was approximately 2741.35 million US$ in 2000, 2616.81 million US$ in 2005, and 2499.06 million US$ in 2010, representing a decrease of 242.29 million US$, or 8.84%, in the decade. Forestland, grassland, wetland, and water bodies played vital roles in the function of ecosystem services, with an aggregated ESV of 99% of the total. The largest proportion of the total ESV was the protection of biodiversity at 48.6%. ESV of all land use types and single ecosystem services continued to decline during 2000–2010. A mechanism of adaptable risk prevention should be established, and unreasonable human activities should be avoided to protect ecosystems and to improve the functions of ecosystem services.     

森林土壤固碳机理研究进展

全球碳平衡中,土壤有机碳储量为2000 Pg,植被碳储量500 Pg,大气碳储量785 Pg,土壤中有机碳变化是影响大气温室气体含量的重要因素。中国人工林总量世界第一,可以通过造林树种的选择,增加森林土壤的碳汇功能,它主要通过4种机理来实现,包括稳定性有机-矿物复合体的形成、持久性封存的深层碳的增加、耐分解有机物成份的积累、以及土壤团聚体结构中碳的物理性保护。中国近年来对木材的需求上升,导致大量短轮伐期人工林的种植,采伐、火烧炼山、施肥与整地等营林措施对土壤碳汇功能形成重大影响。因而,通过加强对中国人工林土壤固碳机理的研究,通过人为措施实现具有较强固碳能力的森林类型,从而提高人工森林生态系统的固碳能力,它对中国减排增汇战略具有重要意义,而加强对不同营林措施对碳汇功能影响的研究是中国当前面临的重大课题。     

Employment of passive sampling in monitoring indoor air quality in selected residences in a Tri-city area in Poland

Time-weighted average concentrations of selected volatile compounds were measured in chosen residences in a Tri-City area of Poland by means of passive sampling. The results were compared to those obtained by dynamic technique – sorption tubes filled with Tenax TA sorbent. Results obtained by employing the two techniques were similar. Total volatile organic compound (TVOC) parameters were also determined. An attempt was also made to evaluate the influence of outdoor air pollutants on indoor air quality.     

中国公众参与环境管理的研究

回顾了中国公众参与环境管理的发展历程,分析了公众参与环境管理存在的问题。针对公众参与环境保护管理的有效性的影响因子,例如主观因素、传统背景、时效因素及执行技巧等进行一系列的探讨,并根据公众参与计划的设计构思、公众参与计划的关键两方面的讨论,提出增加公众参与环境保护管理有效性的一些建议。     

Environmental education in comprehensive schools in Greece

The new institution of comprehensive high school, in Greece today, aims to relate the school with the natural, social, economical and cultural environment. In this frame of process, interrelation and dynamic contact of school with the holistic environment, the subject “Ecology and Environment”; of the 2nd class (16–17 years old), informs, sensitizes and activizes the anxious young generation. The textbook of the subject determines and analyzes the human impact and their consequences to the natural ecosystems. In the total content there are also numerous examples from the natural, man‐made and cultural environment of Greece.     

生物扰动在水层-底栖界面耦合中的作用

综合评述了生物扰动在水层-底栖界面耦合中的作用研究进展和热点问题,并进行了展望。国际上十分重视水层-底栖界面耦合过程的研究并已有一定基础,生物扰动作为海洋生态学的重要内容之一早在20世纪50—60年代就已开展了工作,但直到10余年前才真正开始定量研究,进入了实验模拟、现场观测与建立模型相结合的新阶段。国内的生物扰动研究尚处于起步阶段,加上技术方法落后,一直未能取得突破性进展。目前国内外关于水层-底栖界面生物扰动效应研究基本上都在海洋中开展,尤其在河口、近岸和浅海水域进行,湖泊和河流研究明显薄弱。作为水生态动力学的重要分支和前沿领域,生物扰动研究具有广阔前景。急需针对我国水域特点,引进先进的现场观测和室内测试手段,进一步揭示水层-底栖界面耦合过程中的生物扰动机制,更精准和更详细地掌握水域中各种物质的生物地球化学循环的全过程,为控制水体内源污染释放、富营养化治理和生态建模等提供科学依据。     

Variation in male courtship costs in butterflies

A cost of mating is common to both sexes but has predominantly been examined in females. In species where males provide resources to females at copulation, male mating costs are expected to be high as nutrient provisioning enhancing female fecundity is assumed to carry costs. In addition, males frequently court females prior to mating, which is known to carry survival costs to both sexes. However, the magnitude and basis of variation in males’ mating costs remains largely unknown. Here, I examine the effect of nutrient provisioning and courtship on male longevity across full-sib families in the paternally investing green-veined white butterfly, Pieris napi. Copulating males suffered a survival cost as did courting males prevented from copulating, indicating the courtship component of mating is costly. Male P. napi release aphrodisiacs during courtship to promote mating, indicating that these compounds may also be costly to produce. Contrary to expectation, nutrient provisioning was not associated with reduced survival relative to males only allowed to court females, although it is possible that this could be masked by the potentially elevated courtship rates of courting males relative to mating males. Families differed in magnitude of reduced male survivorship, indicating a likely genetic basis to variation in costs of courtship and copulation. Male weight was unrelated to longevity and mating success, whereas longevity strongly influenced male mating success, indicating lifespan is an important male fitness trait in this species.     

Is the antipredatory response in behaviour reflected in stress measured in faecal corticosteroids in a small rodent?

Predation risk has been shown to alter various behaviours in prey. Risk alters activity, habitat use and foraging, and weight decrease might be a consequence of that. In mammals, studies on physiological measures affected by risk of predation, other than weight, are rare. We studied in two separate laboratory experiments foraging, hoarding behaviour and expression of stress measured non-invasively from the faeces in the bank vole (Clethrionomys glareolus), a common boreal rodent. Voles were exposed to predation risk using odours of the least weasels (Mustela nivalis nivalis). Distilled water served as control. In the first experiment, we found that foraging effort, measured as sunflower seeds taken from seed trays filled with sand, was significantly lower in trays scented with weasel odour. Both immediate consumption of seeds and hoarding were affected negatively by the weasel odour. Females hoarded significantly more than males in autumn. In the second experiment, the negative effect of weasel odour on foraging was consistent over a 3-day experiment, but the strongest effect was observed in the first night. Foraging increased over the time of the experiment, which might reflect either energetic compensation during a longer period of risk, predicted in the predation risk allocation hypothesis, or habituation to the odour-simulated risk. Despite decreased foraging under predation risk, stress measured as corticosteroid metabolite concentration in vole faeces was not affected by the weasel odour treatment. In conclusion, we were able to verify predation-risk-mediated changes in the foraging effort of bank voles but no physiological stress response was measured non-invasively, probably due to great individual variation in secretion of stress hormones.