转基因逃逸及其环境生物安全评价研究进展——抗虫水稻案例分析

转基因技术研发为提高我国水稻产量和减少劳动力投入提供了巨大机遇。我国对转基因水稻研发进行了大量的投入,目前已培育了具有不同新性状的转基因水稻品系,许多品系已进入生物安全评价阶段。风险评价对转基因水稻的安全生产至关重要,是其商品化生产之前必须解决的问题,其中包括转基因逃逸及其潜在环境影响。对水稻抗虫转基因逃逸及其潜在环境风险的评价包括3个重要环节：（1）通过田间试验和模型模拟检测转基因漂移到非转基因栽培稻及其野生近缘种的频率;（2）检测转基因在栽培稻和野生近缘种后代中的表达;（3）确定转基因对野生近缘种群体适合度和进化潜力的影响。大量研究表明,在近距离的空间范围内栽培稻品种之间的基因漂移频率很低（〉0.1%）,但栽培稻与其野生近缘种的基因漂移频率变异很大。进一步研究还表明,Bt抗虫转基因在栽培稻与普通野生稻后代中均能正常表达,但在其不同生长阶段,表达量有很大变异。在有较高水平的害虫虫压下,含有抗虫转基因的栽培稻及野生近缘种杂交后代与不含转基因的对照相比,抗虫性显著提高且适合度利益明显;但是,在虫害发生水平较低时,含有抗虫转基因的群体与不含抗虫转基因的群体相比没有显著的适合度优势。综上,转基因逃逸到非转基因水稻的频率极低,并且可以通过空间隔离阻断其逃逸。虽然抗虫转基因向杂草稻以及与栽培稻距离较近的野生稻群体的逃逸无法避免,但是野生稻和杂草稻群体周围环境中的总体虫压较低,所以基因漂移带来的环境影响应十分有限。     

转基因植物对农业生物多样性的影响

论述了近年来转基因植物对农业生态系统生物多样性影响的研究进展．主要在遗传多样性、物种多样性和生态系统多样性3个层次上予以评述．包括转基因植物对作物遗传多样性的影响;转基因植物的外源基因向杂草和近缘野生种转移;转基因抗虫植物对目标害虫的影响。抗除草剂转基因植物对作物和杂草的影响,抗病毒转基因植物对病毒的影响;转基因植物对非目标生物的影响,对土壤生态系统的影响等．     

转Bt基因水稻对土壤微生态系统的潜在影响

随着转基因作物商品化应用的增多,对其进行生态风险性评价尤为重要.国内外对转基因作物中外源基因向野生亲缘物种漂移的可能性、昆虫对抗虫转基因作物的耐受性以及转基因作物对生物多样性的潜在影响等问题进行了广泛的研究.文中从Bt杀虫结晶蛋白在土壤中的残留特性、Bt杀虫晶体蛋白对土壤微生物可培养类群和土壤酶活性的影响等方面对转Bt基因抗虫水稻的潜在生态风险性进行了简要综述,以期为同类研究提供有益的信息.     

水稻籼、粳花粉源基因飘流的竞争性

基因飘流是水稻(Oryza sativa L.)转基因安全性研究中的重要内容,研究花粉竞争对基因飘流的影响对于建立并完善转基因飘流模型具有重要意义。本实验分别以籼、粳型转基因水稻和籼、粳常规水稻为花粉供体,不育系为花粉受体,研究籼、粳型双供体在基因飘流中的竞争性。结果表明,同种亚种双供体的基因飘流率与花粉源数量成正相关关系,且基因飘流率与供体花粉数量关系符合S曲线。基因飘流是花粉数量竞争和遗传竞争的综合结果。     

转基因的生物安全问题

国家环保总局 2 0 0 2年 5月 2 1日举行国际生物多样性日新闻发布会 ,会议谈到了国际社会目前十分关注的转基因生物及其产品对生物多样性、生态环境和人体健康可能产生的潜在影响 ,其涉及的主要问题有 :1、转基因生物对非目标生物的影响。释放到环境中的抗虫和抗病类转基因植物 ,除对害虫和病菌致毒外 ,对环境中的许多有益生物也将产生直接或间接的不利影响 ,甚至会导致一些有益生物死亡。2、增加目标害虫的抗性和进化速度。研究表明 ,棉铃虫已对转基因抗虫棉产生抗性 ,专家警告 ,如果这种具有转基因抗性的害虫变成对转基因表达蛋白具有抗性…     

盐碱旱地转基因抗虫棉田昆虫群落多样性

棉花是耐干旱和耐盐碱的经济作物。随着土壤的盐碱化和干旱化, 在人口数量和植棉成本剧增的背景下, 我国黄河流域和长江流域棉花种植面积锐减, 棉花种植被迫向滨海盐碱地和内陆及西北干旱地区转移。本文于2013年和2014年在山东东营滨海盐碱地和河北枣强半干旱轻度盐碱地以非转基因棉(‘中棉所49’)为对照, 以转Bt基因棉(‘中棉所79’)为试验材料, 分别作施农药和不施农药处理, 于每年5月初到9月中旬, 调查取样点棉株及地面上害虫及其天敌的种类和数量, 并分析不同施药处理下转基因和非转基因棉田昆虫群落的生物多样性参数差异。结果表明, 施药和不施药转Bt基因棉田昆虫群落和害虫亚群落昆虫的个体总数均低于非转基因棉田, 其中昆虫群落和害虫亚群落个体数在二者之间差异显著; 转Bt基因棉田昆虫群落和害虫亚群落昆虫的多样性指数和均匀度指数均高于非转基因棉田, 而优势集中性指数均低于非转基因棉田, 但差异均不显著。施药条件下两种棉田的昆虫群落和害虫亚群落昆虫个体总数、多样性指数和均匀度指数均低于不施药棉田, 优势集中性指数均高于不施药棉田, 但转基因棉田和非转基因棉田之间无显著差异。表明转基因抗虫棉在盐碱旱地对棉田靶标害虫具有较好的控制作用, 棉田昆虫群落稳定性较高, 昆虫群落对外界的入侵和干扰缓冲能力强, 而化学农药的使用对昆虫群落杀伤力较大, 容易导致某种昆虫的抗性产生和昆虫群落的不稳定, 但比非盐碱旱地棉田昆虫群落生物多样性低, 棉田生态系统更简单。     

转基因抗虫作物对非靶标昆虫的影响

转基因抗虫作物自 1996年被批准商业化种植以来 ,它的抗虫性和经济效益已得到了普遍肯定 ,同时 ,转基因抗虫作物对非靶标生物的影响 ,如转基因抗虫作物的长期种植 ,是否会导致次要害虫上升为主要害虫 ,是否会影响有益昆虫 ,包括重要经济昆虫、捕食性和寄生性天敌以及重要蝶类的种类及种群数量 ,已成为转基因抗虫作物生态风险评估的重要内容。一些研究结果表明 ,转基因抗虫作物在对靶标害虫有效控制的同时 ,一些对杀虫蛋白不敏感的非靶标害虫有加重危害的趋势 ,由于种植转基因抗虫作物 ,减少了化学农药的使用 ,客观上也使非靶标害虫种群数量上升 ,这对转基因抗虫作物害虫综合治理提出了新的要求。靶标害虫数量的减少直接影响了害虫天敌种群数量 ,靶标害虫取食转基因抗虫作物后发育迟缓 ,也间接影响了天敌昆虫的生长发育 ,转基因抗虫作物的花粉或花蜜是一些重要经济昆虫如蜜蜂、熊蜂和一些寄生蜂 ,甚至捕食性天敌的食物来源 ,或花粉飘落到一些鳞翅目昆虫如家蚕或重要蝶类昆虫的寄主植物上 ,直接或间接对这些昆虫造成一定影响。目前大多数研究表明转基因抗虫作物对非靶标昆虫 ,特别是对有益昆虫没有明显的不利影响 ,也有研究报道认为对某些有益昆虫有一定的不良影响。这为深入开展转基因抗虫作物的生态安全     

转Bt抗虫稻对地上非靶标节肢动物的生态风险性

通过转入Bt基因使作物获得抗虫性,减少杀虫剂施用量,为害虫管理开辟了一条重要途径。但公众关注与担忧其环境释放可能存在生物安全与生态风险。本文在系统评述转基因抗虫水稻的生态风险、安全性评价研究内容与方法基础上,重点分析了转基因抗虫水稻对非靶标害虫、捕食性天敌、寄生蜂、中性昆虫及其在食物链上传递的生态风险与安全性,提出了未来研究发展的方向。     

转Cry1Ah基因抗虫玉米HGK60对田间节肢动物及杂草多样性的影响

近年来,随着转基因技术快速发展,转基因作物环境释放风险成为需要重点关注的问题,而对生物多样性的影响研究是客观评价其风险的重要手段。本研究以2018年海南春、冬两季转Cry1Ah基因抗虫玉米‘HGK60'及其对照常规玉米‘郑58'为研究对象,重点探讨了其对田间节肢动物、虫害及杂草多样性的影响。 结果表明: 田间调查共记录节肢动物43753头,隶属7目19科69种,转基因抗虫玉米HGK60与对照常规玉米郑58的节肢动物群落组成、结构及各生态指标均无显著差异;与郑58玉米相比,转基因抗虫玉米HGK60对钻蛀类害虫具有明显的抗性;田间杂草隶属8科16种,转基因抗虫玉米HGK60与郑58玉米田间杂草在密度及各生态指标上差异均不显著,且二者在整个生育期的变化趋势基本一致。说明转Cry1Ah基因抗虫玉米HGK60的种植对田间节肢动物及杂草多样性无显著的影响。该结果为转基因作物的环境风险研究提供了一定的数据支持。     

我国转基因水稻商品化应用的潜在环境生物安全问题

转基因水稻的研发和商品化应用将为提高我国水稻的生产力提供新的机遇,并缓解我国的粮食安全问题.转基凶水稻的人规模环境释放和商品化生产可能会带来一定的环境生物安全问题,处理不好会影响转基因水稻的进一步研究和发展.通常所指的环境生物安全问题主要包括以下几个方面:(1)抗生物胁迫转基因对非靶标生物的影响及效应;(2)外源基因向非转基因作物和野生近缘种逃逸及其可能带来的生态后果;(3)转基因作物对农业生态系统、土壤微生物以及生物多样性的潜在影响;(4)抗生物胁迫转基因的长期使用导致靶标生物对转基因产生抗性等.为了安全有效和持续利用转基因生物技术及其产品,有必要对转基因水稻的环境生物安全性进行科学评价.基于风险评价的原则,本文对转基因水稻在我国商品化生产和大规模种植可能带来的环境生物安全问题进行了理性分析,希望为我国转基因水稻商品化应用的决策和生物安全评价提供科学依据.     

Constitutive expression of a cowpea trypsin inhibitor gene,CpTi, in transgenic rice plants confers resistance to two major rice insect pests

The gene encoding a cowpea trypsin inhibitor (CpTI), which confers insect resistance in trangenic tobacco, was introduced into rice. Expression of the CpTi gene driven by the constitutively active promoter of the rice actin 1 gene (Act1) leads to high-level accumulation of the CpTI protein in transgenic rice plants. Protein extracts from transgenic rice plants exhibit a strong inhibitory activity against bovine trypsin, suggesting that the proteinase inhibitor produced in transgenic rice is functionally active. Small-scale field tests showed that the transgenic rice plants expressing the CpTi gene had significantly increased resistance to two species of rice stem borers, which are major rice insect pests. Our results suggest that the cowpea trypsin inhibitor may be useful for the control of rice insect pests.     

Impact of insect-resistant transgenic rice on target insect pests and non-target arthropods in China

Progress on the research and development of insect-resistant transgenic rice, especially expressing insecticidal proteins from Bacillus thuringiensis （Bt）, in China has been rapid in recent years. A number of insect-resistant transgenic rice lines/varieties have passed restricted and enlarged field testing, and several have been approved for productive testing since 2002 in China, although none was approved for commercial use until 2006. Extensive laboratory and field trials have been conducted for evaluation of the efficiency of transgenic rice on target lepidoteran pests and potential ecological risks on non-target arthropods. The efficacy of a number of transgenic rice lines currently tested in China was excellent for control of the major target insect pests, the rice stem borers （Chilo suppressalis, Scirpophaga incertulas, Sesamia inferens） and leaffolder （ Cnaphalocrocis medinalis）, and was better than most insecticides extensively used by millions of farmers at present in China. No significantly negative or unintended effects of transgenic rice on non-target arthropods were found compared with non-transgenic rice. In contrast, most of the current insecticides used for the control of rice stem borers and leaffolders proved harmful to natural enemies, and some insecticides may directly induce resurgence of rice planthoppers. Studies for developing a proactive insect resistance management of transgenic rice in the future are discussed to ensure the sustainable use of transgenic rice.     

Transgenic plants expressing the AaIT/GNA fusion protein show increased resistance and toxicity to both chewing and sucking pests

The adoption of pest‐resistant transgenic plants to reduce yield losses and decrease pesticide use has been successful. To achieve the goal of controlling both chewing and sucking pests in a given transgenic plant, we generated transgenic tobacco, Arabidopsis, and rice plants expressing the fusion protein, AaIT/GNA, in which an insecticidal scorpion venom neurotoxin (Androctonus australis toxin, AaIT) is fused to snowdrop lectin (Galanthus nivalis agglutinin, GNA). Compared with transgenic tobacco and Arabidopsis plants expressing AaIT or GNA, transgenic plants expressing AaIT/GNA exhibited increased resistance and toxicity to one chewing pest, the cotton bollworm, Helicoverpa armigera. Transgenic tobacco and rice plants expressing AaIT/GNA showed increased resistance and toxicity to two sucking pests, the whitefly, Bemisia tabaci, and the rice brown planthopper, Nilaparvata lugens, respectively. Moreover, in the field, transgenic rice plants expressing AaIT/GNA exhibited a significant improvement in grain yield when infested with N. lugens. This study shows that expressing the AaIT/GNA fusion protein in transgenic plants can be a useful approach for controlling pests, particularly sucking pests which are not susceptible to the toxin in Bt crops.     

转基因水稻外源基因向近缘种群漂流和渐渗的研究进展

水稻是我国最重要的粮食作物之一,我国有8亿以上的人口以稻米作为主食。但在水稻生产中,由于病、虫、草害及不良气候等逆境因子的影响,严重制约了水稻的高产、稳产。转基因生物技术的迅速发展,为水稻抗性育种提供了新途径。自20世纪80年代以来,我国全方位地开展了转基因水稻的研发,目前已经培育出大量的抗病、抗虫、抗除草剂和抗逆的转基因水稻品种,这将为提高我国水稻的生产力和确保粮食安全做出重要的贡献。但转基因水稻的基因漂流及其可能带来的生物安全问题备受关注。已有报道证明,外源转基因可以通过异交向非转基因品种和野生近缘种漂流。在不同的试验条件下,抗除草剂基因有0．05％-0．53％逃逸的可能,其向不育系的最大漂移频率可达4．518％。抗虫基因向相邻非转基因水稻的平均漂移频率最高为0．875％。因此,本文对水稻与其近缘野生种的杂交情况,转基因水稻外源基因向非转基因品种、野生近缘种以及野生非近缘种的漂流和渐渗及其潜在的生态环境风险等方面进行了简要分析,并对转基因水稻的发展进行了展望,以期为转基因水稻的安全应用提供参考。     

田间转Bt基因水稻上稻飞虱卵量、孵化率及天敌作用

为了评估转基因水稻释放后对非靶标害虫及其天敌的影响, 本研究在浙江大学实验农场和中国水稻研究所实验农场2008和2009年连续两年监测了转Bt基因水稻对田间稻飞虱着卵量、卵孵化率、卵被捕食率、卵被寄生率和卵死亡率的影响。结果表明: 两年2个调查地点3个水稻品系（克螟稻1、克螟稻2和秀水11）上的稻飞虱平均每分蘖着卵量、卵孵化率、卵被捕食率和卵被寄生率的趋势基本一致, 而且田间3个水稻品系上稻飞虱卵量的高峰期均持续2周左右; 但是两年2个调查地点3个水稻品系上稻飞虱卵死亡率则没有一致的趋势。此外, 总体上3个水稻品系上稻飞虱平均每分蘖着卵量、卵孵化率、卵被捕食率、卵被寄生率和卵死亡率差异均不显著（P>0.05）。因此, 稻田中转Bt基因水稻对稻飞虱卵没有显著的影响。     

Population-scale laboratory studies of the effect of transgenic plants on nontarget insects

Studies of the effects of insect-resistant transgenic plants on beneficial insects have, to date, concentrated mainly on either small-scale "worst case scenario" laboratory experiments or on field trials. We present a laboratory method using large population cages that represent an intermediate experimental scale, allowing the study of ecological and behavioural interactions between transgenic plants, pests and their natural enemies under more controlled conditions than is possible in the field. Previous studies have also concentrated on natural enemies of lepidopteran and coleopteran target pests. However, natural enemies of other pests, which are not controlled by the transgenic plants, are also potentially exposed to the transgene product when feeding on hosts. The reduction in the use of insecticides on transgenic crops could lead to increasing problems with such nontarget pests, normally controlled by sprays, especially if there are any negative effects of the transgenic plant on their natural enemies. This study tested two lines of insect-resistant transgenic oilseed rape (Brassica napus) for side-effects on the hymenopteran parasitoid Diaeretiella rapae and its aphid host, Myzus persicae. One transgenic line expressed the delta-endotoxin Cry1Ac from Bacillus thuringiensis (Bt) and a second expressed the proteinase inhibitor oryzacystatin I (OC-I) from rice. These transgenic plant lines were developed to provide resistance to lepidopteran and coleopteran pests, respectively. No detrimental effects of the transgenic oilseed rape lines on the ability of the parasitoid to control aphid populations were observed. Adult parasitoid emergence and sex ratio were also not consistently altered on the transgenic oilseed rape lines compared with the wild-type lines.     

Variant cry1Ab entomocidal Bacillus thuringiensis toxin gene facilitates the recovery of an increased number of lepidopteran insect resistant independent rice transformants against yellow stem borer (Scirpophaga incertulus) inflicted damage

The primary technical constraint plant scientists face in generating insect resistant transgenic crops with insecticidal Bacillus thuringiensis (Bt) crystal protein (Cry) genes remains failing to generate sufficiently large numbers of effective resistant transgenic plant lines. One possible means to overcome this challenge is through deployment of a Cry toxin gene that contains high levels of insecticidal specific activity for target insect pests. In the present study, we tested this hypothesis using a natural variant of the Cry1Ab toxin under laboratory conditions that possessed increased insecticidal potency against the yellow stem borer (YSB, Scirpophaga incertulus), one of the most damaging rice insect pests. Following adoption of a stringent selection strategy for YSB resistant transgenic rice lines under field conditions, results showed recovery of a significantly higher number of YSB resistant independent transgenic plant lines with the variant cry1Ab gene relative to transgenic plant lines harbouring cry1Ab berliner gene. Structural homology modelling of the variant toxin peptide with the Cry1Aa toxin molecule, circular dichroism spectral analysis, and hydropathy plot analysis indicated that serine substitution by phenylalanine at amino acid position 223 of the Cry1Ab toxin molecule resulted in a changed role for α-helix 7 in domain I of Cry1Ab for enhanced toxicity.     

Development and field performance of nitrogen use efficient rice lines for Africa

Nitrogen (N) fertilizers are a major input cost in rice production, and its excess application leads to major environmental pollution. Development of rice varieties with improved nitrogen use efficiency (NUE) is essential for sustainable agriculture. Here, we report the results of field evaluations of marker‐free transgenic NERICA4 (New Rice for Africa 4) rice lines overexpressing barley alanine amino transferase (HvAlaAT) under the control of a rice stress‐inducible promoter (pOsAnt1). Field evaluations over three growing seasons and two rice growing ecologies (lowland and upland) revealed that grain yield of pOsAnt1:HvAlaAT transgenic events was significantly higher than sibling nulls and wild‐type controls under different N application rates. Our field results clearly demonstrated that this genetic modification can significantly increase the dry biomass and grain yield compared to controls under limited N supply. Increased yield in transgenic events was correlated with increased tiller and panicle number in the field, and evidence of early establishment of a vigorous root system in hydroponic growth. Our results suggest that expression of the HvAlaAT gene can improve NUE in rice without causing undesirable growth phenotypes. The NUE technology described in this article has the potential to significantly reduce the need for N fertilizer and simultaneously improve food security, augment farm economics and mitigate greenhouse gas emissions from the rice ecosystem.     

Bt水稻对田间非靶标害虫种群动态的影响

以转cry1Ab/cry1Ac基因水稻汕优63(以下简称汕优63/Bt)为材料,亲本汕优63(以下简称汕优63/CK)为对照,在田间自然感虫条件下研究汕优63/Bt对稻田几种非靶标害虫种群消长动态的影响。结果表明,汕优63/Bt上稻苞虫Pelopidas mathias(Fabricius)和稻眉眼蝶Mycalesis gotamaMoore幼虫数量都显著低于对照品种,但汕优63/Bt对稻苞虫的毒性强于稻眉眼蝶。汕优63/Bt上稻飞虱(白背飞虱Sogatellafurcifera(Horvath)和褐飞虱Nilaparvatalugens(Stal))混合种群数量通常与对照品种基本一致,在少数调查时间显著高于对照品种。相反,除少数调查时间与对照品种没有显著差异外,稻叶蝉(黑尾叶蝉Nephotettixcincticeps(Uhler)和二点黑尾叶蝉N.viriscens(Distant))混合种群数量通常显著高于对照品种。稻飞虱和叶蝉成为汕优63/Bt上的主要害虫。     

Impacts of nitrogen fertilizer on major insect pests and their predators in transgenic Bt rice lines T2A‐1 and T1C‐19

Nitrogen is a critical factor for plant development and nitrogen input is one of the important tactics to enhance the development and yield of crops. Nevertheless, nitrogen input could influence the occurrence of insects positively or negatively. Nitrogen is also one of the main elements composing the insecticidal crystal (Cry) protein. Cry protein production could affect nitrogen partitioning in Bt plants and as such nitrogen input may influence insect pest management in transgenic Bt rice, Oryza sativa L. (Poaceae). To test this possibility, we evaluated the impacts of nitrogen regimes on the main insect pests and their predators on two Bt rice lines, T2A‐1 and T1C‐19, expressing Cry2A and Cry1C, respectively, and their non‐transgenic parental counterpart MH63. The results showed that Cry proteins with different nitrogen regimes have enough insecticidal activity on rice leaffolder, Cnaphalocrocis medinalis Guenée (Lepidoptera: Crambidae), in both laboratory and field experiments. Laboratory studies indicated that relevant parameters of ecological fitness in brown planthopper, Nilaparvata lugens (Stål) (Hemiptera: Delphacidae), a non‐target insect pest, were significantly affected by nitrogen input both on Bt and MH63 rice lines. Nymphal survival, female adult longevity, and egg hatchability in N. lugens differed significantly among rice varieties. The experiments conducted in rice fields also demonstrated that nitrogen was positively correlated with the abundance of N. lugens on Bt rice, similar to that on MH63 rice. The abundances of two predators – the wolf spider Pirata subpiraticus (Boesenberg & Strand) (Araneae: Lycosidae) and the bug Cyrthorhinus lividipennis Reuter (Hemiptera: Miridae) – were significantly affected by rice growth stages but not by nitrogen input and rice varieties. In conclusion, the above results indicate that high nitrogen regimes for Bt rice (T2A‐1 and T1C‐19) and non‐Bt rice (MH63) cannot facilitate the management of insect pests.