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1.
Dietary exposure studies are initial steps in environmental risk assessments of genetically engineered plants on non‐target organisms. These studies are conducted in the laboratory where surrogate species are exposed to purified and biologically active insecticidal compounds at higher concentrations than those expected to occur in transgenic crops foliage. Thus, dietary exposure (early tier) tests provide robust data needed to make general conclusions about the susceptibility of the surrogate species to the test substance. For this, we developed suitable artificial diet and used it to establish a dietary exposure test for assessing the toxicity of midgut‐active insecticidal compounds to the larvae of the Asian ladybird beetle Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae). Using boric acid as a model compound, we validated the bioassay established for H. axyridis larvae. An artificial diet containing boric acid which negatively affected survival, development and adult weights was offered to larvae and indicated that the bioassay was able to detect toxic effects of insecticidal substances incorporated in diets. Using this dietary exposure test, environmental risk assessment of Cry1Ac, Cry2Ab, Cry1Ca, Cry1F and the non‐Cry protein Vip3Aa was evaluated by analysing pupation rates, adult emergence rates, 7‐day larval weights, and freshly emerged male and female weights among the toxin treatments and a pure artificial diet. These life‐table parameters did not vary among artificial diets containing 200 μg/g Bt proteins or pure artificial diet. In contrast, boric acid adversely affected all life‐table parameters. Thus on these bases, we concluded H. axyridis larvae are not sensitive to these Bt proteins expressed in genetically engineered crops.  相似文献   

2.
The application of recombinant DNA technology has resulted in many insect-resistant varieties by genetic engineering (GE). Crops expressing Cry toxins derived from Bacillus thuringiensis (Bt) have been planted worldwide, and are an effective tool for pest control. However, one ecological concern regarding the potential effects of insect-resistant GE plants on non-target organisms (NTOs) has been continually debated. In the present study, we briefly summarize the data regarding the development and commercial use of transgenic Bt varieties, elaborate on the procedure and methods for assessing the non-target effects of insect-resistant GE plants, and synthetically analyze the related research results, mostly those published between 2005 and 2010. A mass of laboratory and field studies have shown that the currently available Bt crops have no direct detrimental effects on NTOs due to their narrow spectrum of activity, and Bt crops are increasing the abundance of some beneficial insects and improving the natural control of specific pests. The use of Bt crops, such as Bt maize and Bt cotton, results in significant reductions of insecticide application and clear benefits on the environment and farmer health. Consequently, Bt crops can be a useful component of integrated pest management systems to protect the crop from targeted pests.  相似文献   

3.
谢明  张艳军  吴刚 《昆虫知识》2012,49(4):946-950
利用大蜡螟Galleria mellonella L.诱菌法对转Cry1Ac抗虫棉GK-12及其亲本棉花SM-3的根系区域土壤的虫生真菌种类和数量进行监测,评价转Cry1Ac基因对棉田土壤虫生真菌的影响。实验结果显示:检测到的虫生真菌种类,在转基因棉GK-12及其亲本棉SM-3之间无显著差异,均为球孢白僵菌(Beauveria bassiana)和金龟子绿僵菌(Metarhizium anisopliae);在棉花开花初期、盛花期、结铃期或吐絮期,土壤虫生真菌数量在抗虫棉GK-12和亲本SM-3之间存在显著的差异,其他生育期无显著差异;在距棉花主根不同距离(5 cm\15 cm\25 cm)的区域,土壤虫生真菌数量和种类在抗虫棉GK-12和亲本SM-3之间无显著差异。  相似文献   

4.
With the cultivation of Bt cotton, the produced insecticidal Cry proteins are ingested by herbivores and potentially transferred along the food chain to natural enemies, such as predators. In laboratory experiments with Bollgard II cotton, concentrations of Cry1Ac and Cry2Ab were measured in Lepidoptera larvae (Spodoptera littoralis, Heliothis virescens), plant bugs (Euschistus heros), aphids (Aphis gossypii), whiteflies (Bemisia tabaci), thrips (Thrips tabaci, Frankliniella occidentalis), and spider mites (Tetranychus urticae). Tritrophic experiments were conducted with caterpillars of S. littoralis as prey and larvae of ladybird beetles (Harmonia axyridis, Adalia bipunctata) and lacewings (Chrysoperla carnea) as predators. Immunological measurements (ELISA) indicated that herbivores feeding on Bt cotton contained 5%–50% of the Bt protein concentrations in leaves except whiteflies and aphids, which contained no or only traces of Bt protein, and spider mites, which contained 7 times more Cry1Ac than leaves. Similarly, predators contained 1%–30% of the Cry protein concentration in prey. For the nontarget risk assessment, this indicates that Bt protein concentrations decrease considerably from one trophic level to the next in the food web, except for spider mites that contain Bt protein concentrations higher than those measured in the leaves. Exposure of phloem sucking hemipterans is negligible.  相似文献   

5.
Abstract.  1. Changes in the frequency of Cry1Ac resistance genes and shifts in tolerance of cotton bollworm, Helicoverpa armigera , to the Cry1Ac toxin were assessed using bioassays of F1 and F2 offspring of isofemale lines from Anci County of Hebei Province (a multiple-crop system including corn, soybean, peanut, and Bt cotton) and Xiajin County of Shandong Province (an intensive Bt cotton planting area) in Northern China during 2002–2005.
2. A conservative analysis of the overall results indicated that there was a small increase in the frequency of major, non-recessive resistance genes over time.
3. The relative average development ratings [RADR – growth rate of a line on a Bt diet in proportion to the growth rate on a non-Bt (NBT) diet] of the bollworm larvae in F1 tests increased significantly from year to year, indicating a gradual trend towards higher tolerance to Cry1Ac in the field populations.
4. There were also significant positive correlations between RADR of the lines in the F1 generation and the RADR of their F2 offspring, indicating that the tolerance was genetically based.
5. Quantitative genetic simulation analysis showed that resistance of H . armigera to Bt cotton in Xiajin could evolve to a high level in 11–15 years if no effective resistance management measures are carried out.  相似文献   

6.
Abstract The effect of transgenic double genes, Cry1A + CpTI cotton and Cry1Ac toxin on the parasitoid, Campoketis chlorideae Uchida of cotton bollworm, Helicoverpa armigera (Hübner), was investigated in the laboratory. Helicoverpa armigera larvae when in the first, second and third instar could not survive if fed on transgenic cotton leaves. Consequently, C. chlorideae larvae could not complete their development if parasitizing on such hosts. After H. armigera larvae were reared on transgenic or traditional cotton leaves for 12J48 hours, they were parasitized by C. chlorideae females. Parasitized larvae continued to feed on transgenic or traditional cotton for 12–48 h. The present results showed that the body weight of larvae of the parasitoids were significantly reduced when parasitized hosts fed on transgenic cotton leaves compared to those fed on traditional cotton. Duration of egg and larvae stage were significantly prolonged, pupal and adult weight of C. chloridae was decreased when the host larvae fed on transgenic cotton leaves longer than 48 h. The development duration of C. chlorideae pupae on the hosts fed on transgenic cotton leaves in each treatment was not significantly different from those of controls. The longevity of parasitoid females and males fed with a solution containing Cry1Ac toxin was not significantly different with that of the control.  相似文献   

7.
1. Studies have shown that Cry proteins of the bacterium Bacillus thuringiensis expressed in transgenic plants can be acquired by nontarget herbivores and predators. A series of studies under field and controlled conditions was conducted to investigate the extent to which Cry1Ac protein from Bt transgenic cotton reaches the third trophic level and to measure the amount of protein that herbivores can acquire and expose to predators. 2. Levels of Cry1Ac in Bt cotton leaves decreased over the season. Among herbivores (four species), Cry1Ac was detected in lepidopteran larvae and the amount varied between species. Among predators (seven species), Cry1Ac was detected in Podisus maculiventris and Chrysoperla rufilabris. 3. In the greenhouse, only 14% of the Cry1Ac detected in the prey (Spodoptera exigua larvae) was subsequently found in the predator P. maculiventris. Detection of Cry1Ac protein in Orius insidiosus, Geocoris punctipes and Nabis roseipennis was probably limited by the amount of prey consumed that had fed on Bt cotton. 4. Purified Cry1Ac was acquired by the small predatory bug G. punctipes but at much higher concentration than found in plants or in lepidopteran larvae. 5. Bt protein was shown to move through prey to the third trophic level. Predatory heteropterans acquired Cry1Ac from prey fed Bt cotton, but acquisition was dependent on the concentration of Cry1Ac conveyed by the prey and the amount of prey consumed. The type and availability of prey capable of acquiring the protein, coupled with the generalist feeding behaviour of the most common predators in the cotton ecosystem, probably constrain the flow of Cry1Ac through trophic levels.  相似文献   

8.
转Cry1Ac+Cry2Ab基因棉对棉蚜生命表参数及种群动态的影响   总被引:1,自引:0,他引:1  
为研究新型转Cry1Ac+Cry2Ab基因棉对棉蚜Aphis gossypii Glover生命表参数及种群动态的影响。2010—2011年以常规棉中棉所49为对照,对新型转Cry1Ac+Cry2Ab基因棉在室内进行了生物测定和田间进行了系统的调查。结果表明,和常规棉相比,转Cry1Ac+Cry2Ab基因棉花上棉蚜的净增值率降低81.69%,差异达显著水平;内禀增长率和周限增长率分别降低65.00%和13.01%,但差异不显著;平均世代周期和种群加倍时间分别增加5.54%和154.19%,后者差异达显著水平。和常规棉相比,2010年转Cry1Ac+Cry2Ab基因棉花百株苗蚜、伏蚜和秋蚜的数量分别降低10.79%、37.18%和17.49%,差异均未达显著水平;2011年转Cry1Ac+Cry2Ab基因棉花百株苗蚜的数量增加2.03%,伏蚜和秋蚜的数量分别降低37.41%和64.03%,差异均未达显著水平。  相似文献   

9.
Post‐market monitoring (PMM) consistent with Swiss and European Union legislation should ensure the detection and prevention of adverse effects on the environment possibly deriving from commercial cultivation of genetically modified (GM) crops. Insect‐resistant GM crops (such as Bt‐maize) raise particular questions regarding disturbances of biological control functions provided by beneficial insects such as predators and parasitoids (so‐called natural enemies). Consensus among regulators, scientists and the agricultural biotech industry on appropriate PMM plans allowing the detection and possibly prevention of such adverse effects is still lacking. The aims of this study were to identify the necessity for PMM of Bt‐maize expressing Cry1Ab on natural enemies and to develop an appropriate PMM plan. The approach chosen consisted in determining what type of monitoring is most appropriate to address potential effects of Bt‐maize on natural enemies during commercial cultivation. This included identifying whether there remain substantial scientific uncertainties that would support case‐specific monitoring. Existing pre‐market risk assessment data indicate that Bt‐maize (Cry1Ab) comprises a negligible risk for disturbances in biological control functions of natural enemies. As a consequence, a faunistic monitoring of specific groups of natural enemies is not considered an appropriate approach to detect failures in biological control functions. Alternatively, an approach is proposed that consists in indirectly analysing biological control functions by surveying outbreaks of maize herbivores. Unusual herbivore outbreaks could indicate failures in biological control functions of natural enemies. Data could be collected via questionnaires addressed to farmers growing Bt‐maize. Significant correlations between unusual occurrences of specific maize herbivores and the cultivation of Bt‐maize would subsequently need specific studies to determine possible causalities in more detail. The here proposed approach has the advantage of covering different natural enemy groups. It represents a cost‐effective strategy to obtain scientifically sound data as a basis for regulatory decision‐making.  相似文献   

10.
【背景】转基因棉花在商业化种植之前,必须评价其环境安全性。其中新型棉花材料的生存竞争能力和对物种丰富度的影响是评价的重要内容。【方法】以转Cry1Ac+Cry2Ab基因棉为试验材料,转Cry1Ac棉花中棉所41和非转基因棉花中棉所49为对照品种,分别于2014年5~9月对棉花株高、主茎叶片数、叶绿素含量、比叶面积、果枝数、蕾铃数等生长参数进行比较,同时对二代、三代和四代棉铃虫发生期棉田物种丰富度进行系统调查。【结果】转Cry1Ac+Cry2Ab棉花的生长势与转Cry1Ac棉花和非转基因棉花基本相当,没有表现出明显的竞争优势;产量构成参数在成铃和脱落等方面比非转基因棉表现出良好的优势。对棉田节肢动物物种丰富度的影响表明,转Cry1Ac+Cry2Ab棉花对靶标害虫棉铃虫具有良好的控制效果,对主要刺吸性害虫棉蚜、棉蓟马、烟粉虱、绿盲蝽与天敌龟纹瓢虫、草间小黑蛛、草蛉和小花蝽等的种群丰富度在个别时期有所影响,但总体上与转Cry1Ac棉田和非转基因棉田没有显著性差异。【结论与意义】转Cry1Ac+Cry2Ab棉花无竞争优势,但目标性状优势较好;对棉田节肢动物物种丰富度无明显影响。研究结果为新型转Cry1Ac+Cry2Ab棉花对棉田环境安全方面的研究进一步补充了内容,为转基因棉花的环境安全评价提供科学数据。  相似文献   

11.
Four genetically modified (GM) maize (Zea mays L.) hybrids (coleopteran resistant, coleopteran and lepidopteran resistant, lepidopteran resistant and herbicide tolerant, coleopteran and herbicide tolerant) and its non‐GM control maize stands were tested to compare the functional diversity of arthropods and to determine whether genetic modifications alter the structure of arthropods food webs. A total number of 399,239 arthropod individuals were used for analyses. The trophic groups’ number and the links between them indicated that neither the higher magnitude of Bt toxins (included resistance against insect, and against both insects and glyphosate) nor the extra glyphosate treatment changed the structure of food webs. However, differences in the average trophic links/trophic groups were detected between GM and non‐GM food webs for herbivore groups and plants. Also, differences in characteristic path lengths between GM and non‐GM food webs for herbivores were observed. Food webs parameterized based on 2‐year in‐field assessments, and their properties can be considered a useful and simple tool to evaluate the effects of Bt toxins on non‐target organisms.  相似文献   

12.
文章以转Cry1Ac基因棉(中棉所41)和常规棉(中棉所49)为对照,研究了转Cry1Ac+Cry2Ab基因棉(639020)在棉花生长的关键时期——蕾期(二代棉铃虫发生期)、花期(三代棉铃虫发生期)和花铃期(四代棉铃虫发生期)对棉铃虫的控制作用,同时研究了639020棉田主要捕食性天敌(中华草蛉幼虫、龟纹瓢虫、小花蝽和草间小黑蛛)对烟粉虱的捕食功能,明确了639020棉花在生长的关键时期对棉铃虫的控制效果及对棉田主要捕食性天敌捕食功能反应的影响。结果表明,639020棉花对二代和三代棉铃虫具有良好的控制作用,抗虫性分别比中棉所41提高了52.85%和16.22%,其中前者差异达显著水平,后者差异不显著。在棉花蕾期、花期和花铃期,639020棉田棉铃虫落卵量都比中棉所41棉田和中棉所49棉田低(除二代棉铃虫发生期);棉铃虫幼虫数量都极显著低于常规棉,且都低于防治指标,但与中棉所41棉田无显著差异。639020棉田中华草蛉、龟纹瓢虫、小花蝽和草间小黑蛛对烟粉虱的捕食功能与中棉所41棉田和常规棉田相比无显著变化。研究结果以期为新型转基因棉花环境安全性研究及其外源基因的抗虫遗传效应和生产应用前景进行安全性评价。  相似文献   

13.
在大田栽培条件下,以转Bt基因抗虫棉GK-12和常规棉泗棉3号为材料,在棉花的苗期、现蕾期、花铃期和吐絮期分别测定棉花叶面细菌、真菌和放线菌的数量变化,并在花铃期和吐絮期对叶面细菌生理群的数量和多样性进行分析.结果表明:棉花叶面可培养微生物数量与其生长发育呈正相关,叶面可培养微生物的数量一般由苗期开始增多,到花铃期达最高峰,吐絮期明显减少;花铃期转Bt基因抗虫棉叶面细菌生理群Simpson指数、Shannon-Wiener指数和均匀度比常规棉升高,吐絮期比常规棉下降.  相似文献   

14.
转Bt基因抗虫棉的生态风险及治理对策   总被引:9,自引:3,他引:9  
评述了转Bt基因抗虫棉的生态风险及治理对策。其生态风险主要表现在目标害虫的抗性和对非目标生物群落的变化。目标害虫与转基因抗虫棉的互相作用和抗虫棉杀虫毒素的时空表达方式是目标害虫抗性发展的主要途径。在转基因抗虫棉田中,虽然对目标害虫的防治次数大为减少,但害虫和天敌群落的稳定性仍不如常规棉田,某种次要害虫大发生的可能性较大。认为将转基因抗虫棉纳入综合防治体系并培育更加高效的抗虫棉是治理目标害虫抗性和防止次要害虫上升的重要措施。  相似文献   

15.
Cotton‐ and maize‐producing insecticidal crystal (Cry) proteins from the bacterium, Bacillus thuringiensis (Bt), have been commercialized since 1996. Bt plants are subjected to environmental risk assessments for non‐target organisms, including natural enemies that suppress pest populations. Here, we used Cry1F‐resistant Spodoptera frugiperda (J.E. Smith) and Cry1Ac and Cry2Ab‐resistant Trichoplusia ni (Hübner) as prey for the assassin bug, Zelus renardii (Kolenati), a common predator in maize and cotton fields. In tritrophic studies, we assessed several fitness parameters of Z. renardii when it fed on resistant S. frugiperda that had fed on Bt maize expressing Cry1F or on resistant T. ni that had fed on Bt cotton expressing Cry1Ac and Cry2Ab. Survival, nymphal duration, adult weight, adult longevity and female fecundity of Z. renardii were not different when they were fed resistant‐prey larvae (S. frugiperda or T. ni) reared on either a Bt crop or respective non‐Bt crops. ELISA tests demonstrated that the Cry proteins were present in the plant at the highest levels, at lower levels in the prey and at the lowest levels in the predator. While Z. renardii was exposed to Cry1F and Cry1Ac and Cry2Ab when it fed on hosts that consumed Bt‐transgenic plants, the proteins did not affect important fitness parameters in this common and important predator.  相似文献   

16.
转Bt基因抗虫棉根际微生物区系和细菌生理群多样性的变化   总被引:51,自引:2,他引:51  
在大田栽培条件下 ,以转 Bt基因抗虫棉 GK-12和常规棉花泗棉 3号作为材料 ,在棉花不同发育时期 ,于 2 0 0 1和 2 0 0 2连续两年测定棉花根际土壤细菌、放线菌和真菌数量的变化 ,并在 2 0 0 2年棉花的花铃期和吐絮期对根际细菌生理群的数量和多样性进行了分析 ,结果表明 :虽然不同年份和生育期棉花根际微生物数量存在差异 ,但是 ,年度间和相同的发育时期棉花根际微生物的数量变化趋势一致。在棉花的苗期和吐絮期 ,转 Bt基因抗虫棉根际微生物的数量与对照差异不显著 ;在棉花的花铃期 ,转 Bt基因抗虫棉根际细菌的数量比对照增加 ,放线菌的数量差异不显著 ,而真菌的数量变化没有规律。在棉花发育的花铃期和吐絮期 ,Bt棉根际细菌生理群的总数量比常规棉增加 ,但是根际细菌生理群的 Simpson指数、Shannon-Wiener指数和细菌生理群分布的均匀度下降  相似文献   

17.
转基因棉花Bt毒蛋白的表达及其生态学效应   总被引:9,自引:0,他引:9  
张少燕  谢宝瑜 《昆虫知识》2002,39(5):328-335
苏云金杆菌毒蛋白 (Bacillusthuringiensisgtoxicprotein)基因导入棉花植株后获得的转Bt棉可以特异性地毒杀棉铃虫及鳞翅目的一些其它害虫 ,有效地保护棉花植株不受此类棉花害虫的危害。Bt毒蛋白在棉花植株中的表达受一些内外界因素的影响而呈明显的时空变化。转Bt棉除了严重影响靶标害虫自身外 ,还能对其它一些非靶标昆虫和环境产生影响。另外 ,该文还对害虫对Bt棉的抗性以及防止害虫产生抗性的治理对策进行了综述  相似文献   

18.
Laboratory feeding experiments were carried out to study prey-mediated effects of artificial diet containing Bacillus thuringiensis proteins on immature Chrysoperla carnea. Activated Cry1Ab toxin and the protoxins of Cry1Ab and Cry2A were mixed into standard meridic diet for Spodoptera littoralis (Boisduval) larvae at the following concentrations; for Cry1Ab toxin, 25, 50, 100 g g–1 diet were used; for Cry1Ab protoxin, the concentration was doubled (50 g g–1 diet, 100 g g–1 diet and 200 g g–1 diet) to give relative comparable levels of toxin concentration. Cry2A protoxin was incorporated into the meridic diet at one concentration only (100 g g–1 diet). For the untreated control, the equivalent amount of double distilled water was added to the meridic diet. Individual C. carnea larvae were raised on S. littoralis larvae fed with one of the respective treated meridic diets described above. The objectives were to quantify and compare the resulting effects on mortality and development time of C. carnea with those observed in two previous studies investigating prey-mediated effects of transgenic Cry1Ab toxin-producing corn plants and the other studying effects of Cry1Ab toxin fed directly to C. carnea larvae. Mean total immature mortality for chrysopid larvae reared on B. thuringiensis-fed prey was always significantly higher than in the control (26%). Total immature mortality of C. carnea reared on Cry1Ab toxin 100 g g–1 diet-fed prey was highest (78%) and declined with decreasing toxin concentration. Cry1Ab protoxin-exposed C. carnea larvae did not exhibit a dose response. Prey-mediated total mortality of Cry1Ab protoxin-exposed chrysopid larvae was intermediate (46–62%) to Cry1Ab toxin exposed (55–78%) and Cry2A protoxin (47%) exposed C. carnea. In agreement with the previous studies, total development time of C. carnea was not consistently, significantly affected by the Bt-treatments except at the highest Cry1Ab toxin concentration. However, both highest mortality and delayed development of immature C. carnea raised on Cry1Ab toxin 100 g g–1 diet – fed prey may have been confounded with an increased intoxication of S. littoralis larvae that was observed at that concentration. At all other B. thuringiensis protein concentrations S. littoralis was not lethally affected. Comparative analysis of the results of this study with those of the two previous studies revealed that in addition to prey/herbivore by B. thuringiensis interactions, also prey/herbivore by plant interactions exist that contribute to the observed toxicity of B. thuringiensis – fed S. littoralis larvae for C. carnea. These findings demonstrate that tritrophic level studies are necessary to assess the long-term compatibility of insecticidal plants with important natural enemies.  相似文献   

19.
Protease inhibitors expressed in transgenic plants can provide enhanced levels of resistance to important pest species. A sequential approach for testing the effects of protease inhibitor-expressing crops on nontarget herbivorous insects has been developed. The approach consists of five tiers. The first two tiers comprise the selection phase. In tier one, field surveys are used to characterise the nontarget invertebrate fauna of a crop. In tier 2, histochemical assays are used to identify the subset of herbivores with a particular class of digestive proteolytic enzymes. In the assessment phase a combination of laboratory worst-case scenario studies (tier 3) and controlled environment or small-scale field trials (tier 4) are used to evaluate the impact of the protease inhibitor-expressing plants on the selected nontarget species. In the final tier, field trials are used to compare the relative effect of transgenic plants and current management practices, such as pesticide use, on selected species. The first four tiers of the approach are described using potatoes expressing cystatins, a family of cysteine proteinase inhibitors, as an example. Although the plants have enhanced levels of resistance to potato cyst nematodes (PCN), Globodera pallida and Globodera rostochiensis, the results establish that they have negligible impact on the nontarget herbivorous insect, Eupteryx aurata.  相似文献   

20.
Abstract 1 Coleomegilla maculata DeGeer feeds on corn pollen in the field, but the degree to which this predator relies on corn pollen as part of its diet is not well understood. We quantified the amount of pollen consumed by C. maculata second, third and fourth instars and adults in the field. 2 Laboratory experiments were conducted to determine the digestion rate and duration of different stadia or stages using temperature regimens that reflected field conditions during anthesis. Coleomegilla maculata larvae and adults were collected from the field and the amount of pollen in their digestive tracts was determined gravimetrically. The rate of digestion, duration of each life stage and the field observations were used to estimate the amount of pollen consumed by second, third and fourth instars and adults. 3 Our models estimate that larvae consume 0.66, 1.67 and 3.30 mg of pollen during the second, third and fourth stadia, respectively. Adults consumed an estimated 13.15 mg during anthesis. 4 The relevance of our results to ecological risk assessment of transgenic insecticidal corn and predator life history strategies is discussed. The results presented here are a first attempt to quantify pollen consumption by a predator, and future areas of research are suggested.  相似文献   

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