Different exposure methods to neonicotinoids influenced biochemical characteristics in cotton aphid,Aphis gossypii Glover (Hemiptera:Aphididae)

We measured lipid, carbohydrate and protein content in three strains of cotton aphid, Aphis gossypii (Hemiptera: Aphididae) from very resistant to neonicotinoids up to semi sensitive and sensitive strains in different exposure method of spraying to neonicotinoids. We observed changes in energy source rates at which each substrate was metabolised under starvation, selection and residue stress method spraying that assist in metabolisation of their biochemical parameters. These particular exposure methods influenced some of biochemical parameters in cotton aphid. Results indicated that Sugar, glycogen, total protein and lipid in aphids have significant changes. Studies showed that among three strains called Ag-R, Ag-M and Ag-D which have different susceptibilities to neonicotinoids, strain Ag-R was the most tolerated aphids in counter of imidacloprid and thiametoxam, strain Ag-D was somewhat more tolerated to these insecticides and strain Ag-M was the most sensitive strain which has the resistance factor of nearly 890 in starvation method and was different depending on the method of exposure. Among energy sources, the total lipid in susceptible strain were decreasing more than resistant strain, whereas total proteins were increasing in resistant strain compared with sensitive strain. Total Glycogen was affected significantly in the stress condition which caused an increase and was the most resistant strain.     

棉花型和黄瓜型棉蚜(Aphis gossypii Glover)的寄主适应性及转移通道

采用寄主转接建立生命表的方法研究了棉花型和黄瓜型棉蚜对不同寄主植物的适应性,以及两寄主型棉蚜是否可通过中间桥梁寄主实现寄主互换的问题。结果表明,两寄主型棉蚜直接互换寄主后,其存活和繁殖力显著下降,表现为棉花型和黄瓜型棉蚜的净增殖率比在原寄主上分别下降980倍和12倍,平均世代寿命缩短5～12d。两寄主型棉蚜均能利用木槿植物,并且适应性没有显著差异。但是两寄主型棉蚜均不能在车前草和大叶黄杨上存活和繁殖后代。西葫芦作物对棉蚜在木槿、棉花和黄瓜寄主上的相互转移起到了重要的桥梁寄主作用。冬寄主木槿上棉蚜可通过甜瓜或西葫芦转移到黄瓜寄主上,棉花和黄瓜上棉蚜也可通西葫芦作物分别转移到黄瓜和棉花作物上,从而形成棉蚜在不同寄主植物间的相互转移通道,造成为害和病毒病的扩张。     

Demographic parameters of cotton aphid, Aphis gossypii Glover （Homoptera： Aphididae） on five cotton cultivars

Life table parameters of the cotton aphid, Aphis gossypii Glover, were estimated on five cotton cultivars （‘ Sealand＇ ,‘Siokra＇ ,＇ Vararnin＇ ,‘ Bakhtegan＇ and ‘ Sahel＇ ）. Demographic parameters of the cotton aphid were assessed at 27.5 ± 1 ℃, 65% ＋ 10% RH and a photoperiod of 14：10 （L： D）h. The shortest developmental time for the nymphal stages was 5.5 days on ‘Siokra＇ and the longest was 6.1 days on‘ Sealand＇. The highest offspring per female was 29.6 on ‘Sahel＇ and the lowest was 15.3 on ‘Sealand＇. The rm values varied from 0.272 on ‘Sealand＇ to 0.382 （day^-1） on ‘Varamin＇. Jackknife estimates of the A. gossypii parameters on these cultivars indicated the greatest developmental rate and fecundity on ‘ Varamin＇ and the poorest on ‘ Sealand＇ cultivar.     

不同寄主植物与棉蚜酯酶活性的关系

通过测定不同越冬寄主植物棉蚜Aphis gossypii种群α-乙酸萘酯（α-NA）酯酶的活力,结果表明：山东高密棉田棉蚜种群酯酶活力是北京花椒棉蚜酯酶活力的2.4倍,石榴、木槿上棉蚜酯酶活力是花椒棉蚜酯酶活力的1.3-1.5倍,不同寄主植物棉蚜α-NA酯酶和β-NA酯酶的频率分布表明：鼠李、花椒棉蚜的α-NA酯酶频率分布范围较集中,酯酶活力（每0.033头蚜虫的OD值）小于0.10时的频率分别为70%和62%,其个体累积率达50%时的酯酶活性（EF50）分别为0.08和0.085(每0.033头蚜虫的OD值）;石榴、木槿和棉苗上α-NA酯酶频率分别为25%、31%和45%,其EF50分别的0.154、0.1368和0.1138,酯酶活力明显比鼠李和花椒棉蚜高;高密棉蚜为4%,EF50为0.2113,频率分布范围较宽。β-NA酯酶的频率分布,鼠李、花椒、木槿和棉苗上棉蚜酯酶活力（每0.033头蚜虫的OD值）小于0.10时为52%-62%的个体,EF50分别为0.098、0.084、0.102和0.091,寄主之间差异不大,而石榴和高密棉蚜分别为23%和3%,EF50分别为0.135和0.2136,与其它4个寄主有明显差异。     

Relationship between susceptibility to neonicotinoids and population dynamics of cotton aphid,Aphis gossypii Glover (Hemiptera: Aphididae)

Efficiency of imidacloprid and thiametoxam on population growth parameters of Aphis gossypii Glover (Hemiptera: Aphididae) was assessed using three bioassay methods including; residual, starvation and FAO dip methods. Aphids were assayed under laboratory conditions at 23 ± 2°C, photoperiod of 16:8 (L:D) and 70% relative humidity. Aphids were transferred to sprayed leaves and Petri dishes in residual and starvation methods, respectively. There was no feeding for the aphids in starvation method. In FAO dip method, insects were dipped for 5 s in pesticide solutions and then transferred to fresh leaves. Results revealed that LC₅₀ values calculated with the starvation and residual methods were respectively 15 and 11% more than FAO dip method (for imidacloprid) and 22 and 18% (for thiametoxam). The LC₅₀ value in starvation method was 3% more than the residual method. Calculated LC₅₀ in starvation and residual methods with imidacloprid respectively caused 160 and 34% increase in intrinsic rates of increase (r _m) and net reproductive rate (R ₀) in comparison with FAO dip method. Generation time (T) and doubling time (DT) were respectively 59 and 62% less than those in FAO dip method. In contrast, thiametoxam (LC₅₀ concentration) in starvation and residual method lead to 9 and 67% increase in r _m and R ₀ parameters compared to FAO dip method. However T and DT were respectively 65 and 92% less than mentioned parameters in FAO dip method. There was not any significant difference between life table parameters calculated by residual and starvation bioassay methods.     

转Bt基因抗虫棉对棉蚜的杀虫剂敏感性及解毒酶系的影响

采用浸叶法,以亲本常规棉作对照,室内测定了吡虫啉和辛硫磷对在转Bt基因棉上取食不同世代棉蚜Aphis gossypii Glover的毒力,并用酶标仪测定了转Bt基因棉对棉蚜酯酶、羧酸酯酶和乙酰胆碱酯酶活力的影响。结果表明,棉蚜对吡虫啉和辛硫磷的敏感性并未受Bt棉的影响,2种杀虫剂对分别取食Bt棉1代和多代（60代以上）的棉蚜与取食亲本常规棉棉蚜的LC₅₀值之间均无显著差异。在抗虫棉GK12上取食1代和21代的棉蚜,其酯酶、羧酸酯酶和乙酰胆碱酯酶的活力与取食亲本常规棉的棉蚜之间差异不显著。可见,转Bt基因棉对棉蚜的杀虫剂敏感性和解毒酶活力无显著影响。     

棉花型和瓜型棉蚜产生有性世代能力的分化

为了明确寄主专化性是否会影响棉蚜的繁殖策略或生活史对策,采用低温和短光照组合（18℃,L∶D=8∶16）分别对棉花型和瓜型棉蚜进行有性世代的诱导,比较两寄主专化型棉蚜产生有翅蚜、雄性母、雌性母及无翅孤雌蚜的能力。结果表明：两寄主专化型棉蚜在产生有性世代能力上存在显著差异,表现为棉花型棉蚜在诱导后代中产生有翅蚜、雄性母和雌性母的比率显著高于瓜型棉蚜,并且雌、雄性母产生的时间明显早于瓜型棉蚜。瓜型棉蚜在诱导条件下产生无翅孤雌蚜的比率显著高于棉花型棉蚜,并且发现有不产生有性世代的营专性孤雌生殖个体,而在棉花型棉蚜中没有发现。产生性母蚜的棉花型和瓜型棉蚜均可同时产生孤雌胎生后代。寄主型与诱导时间长短对棉蚜有性世代的产生存在交互作用。采集于田间棉花和黄瓜上的棉蚜,也表现为棉花上的易产生有性世代,在诱导的第2代中产生性母蚜的比率显著高于黄瓜上的棉蚜,并且性母蚜比率在诱导的第2代与第3代间无显著差异; 但黄瓜上的棉蚜性母蚜产生比率第3代显著高于第2代。由此推测,棉蚜寄主专化性的形成与生活史特性的分化有关。     

Ecological specialization of the aphid Aphis gossypii Glover on cultivated host plants

Many plant-feeding insect species considered to be polyphagous are in fact composed of genetically differentiated sympatric populations that use different hosts and between which gene flow still exists. We studied the population genetic structure of the cotton-melon aphid Aphis gossypii that is considered as one of the most polyphagous aphid species. We used eight microsatellites to analyse the genetic diversity of numerous samples of A. gossypii collected over several years at a large geographical scale on annual crops from different plant families. The number of multilocus genotypes detected was extremely low and the genotypes were found to be associated with host plants. Five host races were unambiguously identified (Cucurbitaceae, cotton, eggplant, potato and chili- or sweet pepper). These host races were dominated by asexual clones. Plant transfer experiments using several specialized clones further confirmed the existence of host-associated trade-offs. Finally, both genetic and experimental data suggested that plants of the genus Hibiscus may be used as refuge for the specialized clones. Resource abundance is discussed as a key factor involved in the process of ecological specialization in A. gossypii.     

Impact of transgenic cotton plants on a non-target pest, Aphis gossypii Glover

Abstract.  1. Transgenic crops have shown great promise for the control of target pest insects, but in some cases they can also influence non-target species. This study investigated the impact of Bt and Bt+CpTI transgenic cottons on the non-target cotton aphid, Aphis gossypii Glover, by comparing life-table parameters, feeding behaviour, and the fluctuating asymmetry of morphological traits of aphids reared on transgenic cotton and those on untransformed control plants.
2. Aphids on the Bt+CpTI cotton showed a shorter reproductive duration and maximum lifespan, lower survival rates and potential maximum fecundity, and an earlier occurrence of peak daily mortality in the first or second generation. However, the aphid population soon developed fitness and overcame the negative effect in the second or third generation. The aphids on the Bt cotton had significantly longer reproductive durations in the first generation, higher survival rates in the third generation, and apparently larger potential maximum fecundity in all three generations.
3. The percentages of accumulated duration of feeding waveforms E1 and E2 were significantly lower in aphids on the Bt+CpTI cotton than in those on the Bt or control cotton, whereas the frequencies of moving and finding feeding sites, and probe behaviour were significantly higher.
4. Fluctuating asymmetry in three morphological characters of aphids reared on transgenic and control cotton was detected. The fluctuating asymmetry value of the third segment of antenna in aphids on Bt+CpTI cotton was significantly higher than that of aphids on Bt or control cotton. Based on the fluctuating asymmetry value, the stress of cotton on the aphids could be ranked as Bt+CpTI cotton > Bt cotton > control cotton.     

棉蚜寄主专化型及其形成的行为机理

通过生活在甜瓜和棉花上的棉蚜Aphisgossypii Glover的行为,研究棉蚜的寄主专化型及其形成的行为机理。生物学观察显示： 两类棉蚜在寄主植物相互交换以后,定居数显著减少,棉花蚜型棉蚜的繁殖系数及若虫存活率显著下降,说明棉蚜存在甜瓜蚜型和棉花蚜型两种寄主专化型。通过刺探电位技术研究棉蚜的取食行为,以探索其寄主专化型形成的行为机理。结果表明： 甜瓜蚜型棉蚜在棉花上的取食行为容易被中断,但其口针定位韧皮部的能力并没有显著削弱;而棉花蚜型棉蚜在甜瓜上的取食行为受到更大的影响,口针无法顺利定位至韧皮部,并在2 h内根本无法在筛管内取食。生物学观察和EPG取食行为分析都显示： 与甜瓜蚜型棉蚜相比,棉花蚜型棉蚜对寄主的要求更严格-寄主专化程度更高,对寄主的利用率更高。     

棉花型和黄瓜型棉蚜对寄主植物的适合度

采用寄主转换建立生命表及触角电位 (EAG)方法比较了棉蚜两寄主专化型 (棉花型和黄瓜型 )对不同夏季作物的适合度。结果表明 ,棉花型蚜转到黄瓜上及黄瓜型蚜转接到棉花上均不能正常产仔繁殖和建立种群。两种蚜型均不能在茄子和苋菜上建立种群。黄瓜型蚜能在豇豆上建立种群 ,但棉花型蚜不能 ,表现出两种寄主型棉蚜对夏寄主利用上存在显著差异。受棉蚜为害 12～ 36 h后的黄瓜或棉花植株仍不适合于棉花型或黄瓜型蚜 ,表现出黄瓜型蚜不能在被棉花型蚜为害过的棉株上正常存活和繁殖 ,棉花型蚜也不能在被黄瓜型蚜为害过的黄瓜苗上存活和繁殖。两种寄主型蚜对不同寄主叶片丙酮提取物的触角电位表明 ,黄瓜型蚜对棉花、哈密瓜和马铃薯叶片提取物的反应显著强于棉花型蚜 ,而对黄瓜和甜瓜叶片提取物的反应上两种蚜型差异不显著。文中同时对棉花型和黄瓜型棉蚜产生的生态机制进行了讨论。     

Symbiotic microbial studies in diverse populations of Aphis gossypii,existing on altered host plants in different localities during different times

Complex interactions between symbiotic bacteria and insects ultimately result in equilibrium in all aspects of life in natural insect populations. In this study, abundance of principal symbiotic bacteria was estimated using qPCR in 1553 individuals of aphids, Aphis gossypii. Aphids were sampled from primary and secondary host plants—hibiscus and cotton. Hibiscus aphids were collected from 24 different locations in April, September, and November, whereas cotton aphids were collected between 2015 and 2017 from areas with wide variations in climatic conditions. About 30%–45% aphids were recorded with the most dominant symbiont, Arsenophonus. The other symbionts were in low frequency, and about 7% of aphids were noted with Hamiltonella, Acinetobacter, and Microbacterium, and 3% of aphids were verified with Serratia and Pseudomonas. Aphids infected with Hamiltonella, Arsenophonus, and Serratia can influence Buchnera densities. Hamiltonella has positive interaction with densities of Arsenophonus and Serratia. Almost 100% coinfection of Hamiltonella and Arsenophonus was detected in Xinxiang aphids and 50% coinfection was reported in aphids from North China, while no coinfection was detected in Hainan aphids. These findings describe the prevalence pattern and richness of core community of symbiotic bacteria in naturally occurring populations of A. gossypii and provide new insights for the study of symbiotic bacteria.     

南京地区棉蚜的飞行活动节律及其飞行能力

昆虫的飞行活动规律及飞行能力是研究其能否迁飞的基础。通过采用春季到秋季20 m高空黄盆诱蚜、高空所诱蚜和春季木槿树上有翅蚜的卵巢解剖,以及春夏秋三季田间有翅蚜的吊飞试验等方法,研究了南京地区棉蚜Aphis gossypii的飞行活动节律和飞行能力。结果表明,有翅棉蚜的日羽化高峰出现在19:00～20:00。2001年南京地区棉蚜的春、秋两季迁飞高峰分别在5月8日和10月25日。5月份高空诱集的棉蚜中,95.7%个体的卵巢小管数在7条以下,而木槿上羽化后1天的有翅蚜中有35.2%个体的卵巢小管数在7条以上;高空诱蚜和木槿上蚜的平均卵巢小管数存在极显著差异,分别为3.94±1.65和5.88±1.92。8月中下旬棉田棉蚜存在低空飞行行为,并且出现飞行高峰时有翅蚜的卵巢小管数平均在6条以下,超过6条则停止飞行。羽化后1～2天有翅棉蚜吊飞个体的飞行比率和平均飞行距离表现为春、秋季显著大于夏季,三季的最长飞行距离分别为3.89 km、6.15 km和1.44 km。     

棉蚜种群季节性变异的生态条件

研究棉蚜在不同温度下取食不同生育期棉叶后种群的变动情况。通过Weibull函数模拟、生命表分析和高氏距离分析比较表明,无论在苗期和蕾期,随着渐度的升高棉蚜种群的死亡高峰日均提前,世代寿命明显缩短;然而,在20-25℃范围内,相同温度时棉蚜生活在棉花蕾期叶上的繁殖力明显高于苗期叶上的繁殖力。在30℃时繁殖力普遍明显下降,温度20-25℃最适于棉蚜的生长与繁殖。且棉花殖生长期比营养生长期更适于棉蚜的生存和繁殖,温度与棉花生育期都是导致苗蚜与伏蚜生长与繁殖差异的外界因素。     

Variation in clonal diversity in glasshouse infestations of the aphid, Aphis gossypii Glover in southern France

Aphis gossypii is an aphid species that is found throughout the world and is extremely polyphagous. It is considered a major pest of cotton and cucurbit species. In Europe, A. gossypii is assumed to reproduce exclusively by apomictic parthenogenesis. The present study investigates the genetic diversity of A. gossypii in a microgeographic, fragmented habitat consisting of eight glasshouses of cucurbit crops. This analysis, which was based on the results from seven microsatellite loci, has confirmed that A. gossypii populations in southern France are primarily asexual, as only 12 nonrecombinant genotypic classes (clones) were identified from 694 aphids. Moreover, a high proportion of the aphids (87%) had one of three common genotypes. No significant correlation was found between genotypic class and host plant species. Within a glasshouse population of A. gossypii, a significant reduction in clonal diversity was observed as the spring/summer season progressed. The final predominance of a clone could result from interclonal competition. At the microgeographic level (i.e. glasshouses within a 500-m radius), significant genetic subdivision was detected and could be attributed to founder effects and the limitation of gene flow imposed by the enclosed nature of the glasshouse structure. Finally, the three common clones of A. gossypii detected in 1996 reappeared in spring 1997 following the winter extinction, together with rare clones that had not previously been seen. The probability that A. gossypii overwinters within refuges at a microgeographic scale from which populations are renewed each spring is discussed.     

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.     

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.