龟纹瓢虫在棉株上对棉蚜的搜索与捕食行为

研究了龟纹瓢虫在棉花上对不同密度、不同空间分布、不同来源棉蚜的搜索与捕食行为.结果表明,龟纹瓢虫对棉蚜的搜索活动随棉蚜密度增加而下降;当棉花上有充足的棉蚜时,瓢虫主要搜索和捕食棉花上部的棉蚜,很少捕食棉花中部的棉蚜,极少捕食下部的棉蚜;瓢虫对棉花不同部位上分布的2种不同来源的棉蚜捕食量没有差异;但当棉花上部主要为美9101棉花棉蚜时,瓢虫的搜索活动明显下降.龟纹瓢虫的捕食行为与蚜虫密度和空间分布密切相关.     

Stylet penetration behaviors of the cotton aphid Aphis gossypii on transgenic Bt cotton

Stylet penetration behaviors of cotton aphids Aphis gossypii Glover on a transgenic cotton line "GK-12" expressing Bt toxic protein of Cry1A (Bt cotton) and a non-Bt conventional cotton line "Simian-3" (CK cotton) were recorded with the direct current electrical penetration graph (DC-EPG) technique. Cotton aphids reared on Bt cotton (abbreviated as Bt-aphids) and its parental non-Bt control line (CK-aphids) for more than 20 generations each, were used for recordings on two cotton lines. Among 47 selected parameters reflecting the activities of aphid stylets within plant tissues, there were eight parameters of CK-aphids showing significant differences between the performances of CK-aphids on Bt cotton and CK cotton, while for Bt-aphids, all the parameters were statistically equal between the performances on the two cotton lines. All parameters with significant differences indicated that CK-aphids could penetrate into Bt cotton more easily, but the phloem saps of Bt cotton were not as good as those of regular cotton for CK-aphids. Based on the present results, we concluded that there were some factors in Bt cotton affecting penetration behaviors of CK-aphids, but it just took several generations for CK-aphids to completely adapt Bt cotton, and Bt-aphids could feed on two cotton lines without difficulty.     

Influence of temperature on bionomics of cotton aphid, Aphis gossypii, on cotton

Life table parameters of Aphis gossypii Glover (Homoptera: Aphididae) on Gossypium hirsutum L. were determined at six temperatures (10, 15, 20, 25, 30, and 35 ± 0.5°C) in the laboratory. Relationships of life table parameters with temperature were described with mathematical equations. Development was fastest at 30°C, with a pre-larviposition period of 4.6 d. Survival to adult was greatest at 25°C (81%). Fecundity was highest at 25°C, with a total fecundity of 28.3 nymphs per female and a mean reproductive rate of 3.1 nymphs per female per day. Threshold temperatures for development in the first through fourth instar and the adult were 8.2, 8.0, 7.2, 6.2 and 7.9°C, respectively. The durations of these stages, expressed as temperature sums above these thresholds, were 24.2, 23.7, 23.0, 25.5 and 168.8 degree-days (D°), respectively. A. gossypii achieved its maximum net reproductive number (24.4 nymphs per female) and greatest intrinsic rate of increase (0.386 d–1) at 25°C. The high relative rate of population increase at 25°C results in a daily population increase of 47% and a doubling time of only 1.8 d, illustrating the tremendous growth capacity of A. gossypii populations under favourable conditions. Compared to literature sources, our source of A. gossypii, fed on cotton, showed a comparatively great heat tolerance.     

Kaolin potential for the integrated management of Aphis gossypii Glov. (Homoptera: Aphididae) on cotton

Kaolin-based particle film has been developed in the United States for use in agriculture as crop protectant. This work is aimed to determine the optimal kaolin concentration for use against Aphis gossypii, the most damaging sucking insect of cotton in Benin, West Africa. Following kaolin concentrations have been tested in four different experiments: 2%, 3%, 4%, 5%, 6% and 8%. The effect of kaolin is tested by recording the population dynamics of A. gossypii on cotton after spraying. Results showed that the solution containing 5% of kaolin has significantly reduced aphids' number. This concentration corresponds to a maximum of 18 kg of kaolin per hectare for the entire season when ultra low volume sprayer is used. Higher kaolin concentrations have left a large deposit of kaolin particle on cotton leaves thus handicapping chlorophyll synthesis and the proper growth and development of the plants. To obtain encouraging results, an adequate application method that provides a full and continuous coverage of the plants with kaolin particle film should be used. We suggest conducting trials on this product in practical cotton production conditions in West Africa to assess its potentiality as integrated pest management (IPM) component.     

短时间高温处理对棉蚜存活的影响

为探索棉蚜Aphis gossypii Glover种群动态与高温的关系,研究了不同温度(30、34、38、42、46℃)、不同处理时间(1、2、5、8h)对棉蚜存活的影响。结果表明:在同一处理时间条件下,棉蚜的存活率随着温度的升高而降低;在同一处理温度下,棉蚜的存活率随处理时间的延长而降低。30～38℃温度范围内,棉蚜存活率降低缓慢,从42℃开始棉蚜的存活率迅速降低。棉蚜半数致死温度随着处理时间的延长而降低;相同处理时间条件下,若蚜半数致死温度高于成蚜,若蚜较成蚜更耐高温。     

棉蚜微卫星DNA的克隆及其多态性检测

微卫星ＤＮＡ (ｍｉｃｒｏｓａｔｅｌｌｉｔｅＤＮＡ)是由 2～ 6ｂｐ核苷酸序列组成的简单串联重复序列 ,广泛存在于基因组的间隔顺序和内含子等非编码区。由于其高度的多态性 ,呈共显性遗传 ,并且其多态性可用ＰＣＲ技术结合电泳被检测出来 ,且可克服ＤＮＡ多位点方法 (如ＲＡＰＤ及ＤＮＡ指纹图等 )中把来自不同位点但具有相同大小的等位基因混淆等优点 ,使其已成为生物群体遗传结构与变异及彼此关系研究中的一种极有价值的分子遗传标记工具。棉蚜Ａｐｈｉｓｇｏｓｓｙｐｉｉ是一种重要的农业害虫 ,其寄主广 ,且季节分化明显。目前已记载…     

Genetic diversity of the cotton aphid Aphis gossypii in the unstable environment of a cotton growing area

1 Spatial and temporal habitat heterogeneity represented by annual crops is a major factor influencing population dynamics of phytophagous insect pests such as the cotton aphid Aphis gossypii Glover. We studied the effects of instability of the cotton agroecosystem resulting from the temporary availability of the plant resource and the repeated use of insecticides on the genetic variability of the cotton aphids.
2 Samples of A. gossypii were collected in cotton plots, treated or not with insecticides and from vegetable crops (Malvaceae, Cucurbitaceae and Solanaceae) within the cotton growing area of northern Cameroon. The genetic structure of the samples was assessed using eight microsatellite markers. Insecticide resistance was estimated through the detection of two mutations in the ace -1 gene that are associated with insensitivity of acetylcholinesterase to carbamate and organophosphate insecticides.
3 The results obtained show that both host plants and insecticides act in genetic structuring of A. gossypii . Ninety-three percent of aphids collected on cotton were characterized by the same microsatellite multilocus genotype, Burk1 , which also displays the insecticide resistant alleles.
4 During the dry season, the cotton crop season after, the genotype Burk1 was principally found on two other malvaceous cultivated plants, rosella and okra, acting as suitable reservoir plants. The ability of the cotton aphid to move among asynchronous suitable habitats in response to changes in resource availability enables the pest to exploit unstable cropping systems. An understanding of the cotton aphid life system may aid to improve strategies for integrated resistance management.     

Potential cotton aphid,Aphis gossypii,population suppression by arthropod predators in upland cotton

The cotton aphid, Aphis gossypii Glover, predation rate of convergent lady beetle, Hippodamia convergens Guerin-Meneville, was determined by assigning a single predator randomly to each of four prey density treatments in the laboratory. Prey densities included 25, 50, 100, and 200 aphids per Petri dish arena. Predation response was recorded at 1, 4, 8, 16, 24, and 48 h after assigning predators to their prey treatments. Rate of consumption increased through time, with all 25 aphids consumed during the first 4 h of the experiment. At the highest density, adult lady beetle consumed on average 49, 99, 131, 163, 183, and 200 aphids within 1, 4, 8, 16, 24 and 48 h, respectively. Predators showed a curvilinear feeding response in relation to total available time, indicating that convergent lady beetles have the potential to suppress larger populations of aphids through continuous feeding by regulating their predation efficiency during feeding. The analysis of age-specific mortality in absence of prey revealed that lady beetles could survive for an extended period of time （more than 2 weeks） without prey. The ability of a predator to survive without prey delays or prevents the rebound of pest populations that is a significant factor in natural biological control. A two-year field sampling of 10 cotton arthropod predator species showed that spiders （27%） were the most dominant foliage dwelling predators in the Texas High Plains cotton followed by convergent lady beetles （23.5%）, hooded beetles （13.5%）, minute pirate bugs （11%）, green lacewings （9.5%）, bigeyed bugs （7.5%）, scymnus beetles （3%）, soft-winged flower beetles （2%）, damsel bugs （1.5%）, and assassin bugs （1.5%）. A field cage study showed that one H. convergens adult per plant released at prey density of one aphid per leaf kept the aphid population below economic threshold for the entire growing season.     

Potential cotton aphid,Aphis gossypii,population suppression by arthropod predators in upland cotton

The cotton aphid, Aphis gossypii Glover, predation rate of convergent lady beetle, Hippodamia convergens Guerin‐Meneville, was determined by assigning a single predator randomly to each of four prey density treatments in the laboratory. Prey densities included 25, 50, 100, and 200 aphids per Petri dish arena. Predation response was recorded at 1, 4, 8, 16, 24, and 48 h after assigning predators to their prey treatments. Rate of consumption increased through time, with all 25 aphids consumed during the first 4 h of the experiment. At the highest density, adult lady beetle consumed on average 49, 99, 131, 163, 183, and 200 aphids within 1, 4, 8, 16, 24 and 48 h, respectively. Predators showed a curvilinear feeding response in relation to total available time, indicating that convergent lady beetles have the potential to suppress larger populations of aphids through continuous feeding by regulating their predation efficiency during feeding. The analysis of age‐specific mortality in absence of prey revealed that lady beetles could survive for an extended period of time (more than 2 weeks) without prey. The ability of a predator to survive without prey delays or prevents the rebound of pest populations that is a significant factor in natural biological control. A two‐year field sampling of 10 cotton arthropod predator species showed that spiders (27%) were the most dominant foliage dwelling predators in the Texas High Plains cotton followed by convergent lady beetles (23.5%), hooded beetles (13.5%), minute pirate bugs (11%), green lacewings (9.5%), bigeyed bugs (7.5%), scymnus beetles (3%), soft‐winged flower beetles (2%), damsel bugs (1.5%), and assassin bugs (1.5%). A field cage study showed that one H. convergens adult per plant released at prey density of one aphid per leaf kept the aphid population below economic threshold for the entire growing season.     

Two different genes encoding acetylcholinesterase existing in cotton aphid (Aphis gossypii).

Two acetylcholinesterase (AChE) genes, Ace1 and Ace2, have been cloned from cotton aphid, Aphis gossypii Glover, using the rapid amplification of cDNA ends (RACE) technique. To the best of our knowledge, this should be the first direct molecular evidence that multiple AChE genes exist in insects. The Ace1 gene was successfully amplified along its full length of 2371 bp. The open reading frame is 2031 bp long and encodes 676 amino acids (GenBank accession No. AF502082). The Ace2 gene was amplified as a mega-fragment of 2130 bp lacking part of 5'-end untranslated region (UTR). The open reading frame is 1992 bp long and ecodes a protein of 664 amino acids (GenBank accession No. AF502081). Both genes have the conserved amino acids and features shared by the AChE family, but share only 35% identity in amino acid sequence. The Ace1 gene is highly homologous to the AChE gene of Schizaphis graminum (AF321574) with 95% identity, and Ace2 to that of Myzus persicae (AF287291) with 92% identity. Phylogenetic analysis showed that the two cloned AChEs of A. gossypii are different in evolution. The phylogenetic tree generated by the PHYLIP program package inferred that AChE2 of A. gossypii is a more ancestral form of AChE. Homology modeling of structures using Torpedo californica (2ACE_) and Drosophila melanogaster (1Q09:A) native acetylcholinesterase structure as main template indicated that the two AChEs of Aphis gossypii might have different three-dimensional structures. Alternative splicing of Ace1 near the 5'-end resulting in two proteins differing by the presence or absence of a fragment of four amino acids is also reported.     

高温和密度效应对棉蚜死亡和繁殖的影响

在室内采用叶子圆片培养基饲养棉蚜的方法,研究不同温度(32、34、36、38、40℃)和密度(每皿5、25、50、75)对棉蚜死亡和繁殖的影响.结果表明:随着温度的升高、密度和持续天数的增加,棉蚜累计死亡率呈上升趋势.互补重对数模型模拟结果表明:β值随着密度的增加而减小,说明随着棉蚜密度的增加,温度对棉蚜的作用减弱;不同持续天数的γj值不同,说明棉蚜条件死亡率受温度和作用时间的共同影响.双因素方差分析结果表明,温度和密度显著影响棉蚜的繁殖率,且存在交互作用.32～36℃范围内,棉蚜繁殖率随密度的增加呈降低趋势,40℃条件下,不同密度棉蚜繁殖率没有差异,说明随着温度的升高,密度对棉蚜的作用逐渐减弱.即在不同的温度区间和密度范围内,温度和密度对棉蚜的作用大小、主次发生了改变.研究结果为棉蚜种群监测、预警和改进害虫控制方法提供了科学依据.     

棉蚜获得黄瓜花叶病毒的行为与取食过程的关系

利用刺吸电位（EPG）及其即时中断技术研究了棉蚜Aphis gossypii传播黄瓜花叶病毒(CMV)的机理,分析电势落差波(pd)及其亚波形在棉蚜获得CMV的取食行为过程中的作用。结果表明：棉蚜的获毒需要pd波的发生,它的获毒效率与穿刺病株细胞膜的次数呈正相关。证明了这种行为是一种细胞内行为。对植物细胞内穿刺产生的电位落差（pd）波亚波形分析的结果显示,棉蚜的获毒发生在pd波的Ⅱ-3阶段,与棉蚜主动吸食植物细胞汁液的活动相关, 这支持了蚜虫获毒的 “吸入假说”。     

Influence of constant temperatures on life history parameters of the cotton aphid, Aphis gossypii, infesting cotton

Laboratory clip-cage studies were conducted to quantify the temperature-dependent development, survivorship, and reproduction and to generate life history characteristics and population growth parameters of the cotton aphid, Aphis gossypii Glover, on phenologically standardized greenhouse-grown cottons at 10, 15, 20, 25, 30, and 35 degrees C. The developmental thresholds were estimated to be 6.3, 6.7, 5.9, 5.9, and 6.3 degrees C for first to fourth instars and for total nymphal development, respectively. The maximum rate of development were estimated to occur at 32.2, 30.8, 30.4, 30.0, and 30.2 degrees C for first to fourth instars and for total nymphal development, respectively. Increased temperature resulted in more rapid decline in survivorship, which was particularly sharp at 35 degrees C, dropping from 94 to 17% in 5 d. Number of days elapsed until first deposition of progeny increased progressively and sharply at temperatures 10 (26 d) to 15 (15 d) to 20 degrees C (8 d) and stabilized at 5 d for 25, 30, and 35 degrees C. Average lifetime fecundity of females rose from a low of 9.76 progeny at 10 degrees C to a peak of 58.9 progeny at 30 degrees C and declined sharply to 17.3 at 35 degrees C. Finite rate of population growth was highest at 25 degrees C and lowest at 10 degrees C. Although stage-specific developmental maxima occurred between 30 and 32 degrees C, a nonlinear regression model estimated 28.6 degrees C to be the optimum temperature for overall cotton aphid development, reproduction, and population increase.     

Purification and characterization of acetylcholinesterase from cotton aphid (Aphis gossypii Glover)

A simple and effective method was set up to purify acetylcholinesterase (AChE, EC3.1.1.7) from the cotton aphid, Aphis gossypii Glover. The procedure involved filtration on a sephadex G-25 column, separation with sephadex G-200 and procainamide affinity column. AChE from both susceptible and resistant strains were purified to a single band as resolved on denaturing polyacrylamide gel electrophoresis (SDS-PAGE). The specific activity increased by 35,100- and 33,680-fold with a yield of 30.3 and 29.8%, respectively. The molecular mass of the purified AChE was about 63,500 Dalton as determined by SDS-PAGE. However, three bands resolved on PAGE gel electrophoresis, leading to the inference that native AChE exists in three forms. The optimum conditions for measuring the activity of purified AChE with kinetic method were 0.02M phosphate buffer, pH7.2, 0.02 mM 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB), and 25 degrees C. Investigation also revealed that crude extract and purified AChE had different kinetic characteristics and inhibitory properties. They responded differently to varied DTNB, ATChI, and phosphate buffer ion concentrations, as well as pH, temperature, and inhibitors. The purified AChE was more sensitive to eserine, methamidophos, and pirimicarb. Especially for resistant aphids, the sensitivity of purified AChE to methamidophos and pirimicarb was enhanced 6.43 and 11.73 times, respectively. We infer that one or more factors in the crude extract from the resistance strain have more influence on AChE sensitivity. Further study is needed to investigate the basis of these observations.     

棉蚜抗氧化乐果品系及敏感品系羧酸酯酶性质的比较

在室内用氧化乐果逐代筛选的棉蚜抗性品系,相对于敏感品系的抗性倍数是17。用α-乙酸萘酯（α-NA）、α-丁酸萘酯（α-NB）、α-磷酸萘酯（α-NP）和β-磷酸萘酯（β-NP）作底物比较研究了氧化乐果抗性和敏感品系棉蚜Aphis gossypii羧酸酯酶的比活力、米氏常数（K_m）和最大反应速度V_max）等有关的动力学常数。以α-NA和α-NB作底物时,抗性品系棉蚜的比活力显著低于敏感品系的;以α-NP和β-NP作底物时,两个品系棉蚜的比活力、K_m和V_max没有明显差异。用α-NA、β-NA作底物染色做酯酶同工酶电泳,抗性品系棉蚜的酯酶同工酶染色比敏感品系棉蚜的浅。     

棉蚜飞行行为与卵巢发育的关系

采用高空黄盆诱蚜、卵巢解剖、体型测量及起飞角度测定方法研究了南京地区棉蚜飞行行为与卵巢发育间的关系。结果表明 ,5月份高空诱蚜的卵巢小管数与个体大小间成显著正相关 ,而木槿上有翅蚜的体长与卵巢小管数间相关不显著。棉蚜起飞角度的大小与卵巢小管数目呈显著的负相关 ,棉蚜的最大起飞角度为 4 4°左右。     

Mutations in acetylcholinesterase associated with insecticide resistance in the cotton aphid, Aphis gossypii Glover

Two acetylcholinesterase genes, Ace1 and Ace2, have been fully cloned and sequenced from both organophosphate-resistant and susceptible clones of cotton aphid. Comparison of both nucleic acid and deduced amino acid sequences revealed considerable nucleotide polymorphisms. Further study found that two mutations occurred consistently in all resistant aphids. The mutation F139L in Ace2 corresponding to F115S in Drosophila acetylcholinesterase might reduce the enzyme sensitivity and result in insecticide resistance. The other mutation A302S in Ace1 abutting the conserved catalytic triad might affect the activity and insecticide sensitivity of the enzyme. Phylogenetic analysis showed that insect acetylcholinesterases fall into two subgroups, of which Ace1 is the paralogous gene whereas Ace2 is the orthologous gene of Drosophila AChE. Both subgroups contain resistance-associated AChE genes. To avoid confusion in the future work, a nomenclature of insect AChE is also suggested in the paper.