Resistance development,cross-resistance,and fitness costs associated with Aphis gossypii resistance towards sulfoxaflor and acetamiprid in different geographical regions

Aphis gossypii has become increasingly difficult to manage due to its strong insecticide resistance.In the laboratory,we established sulfoxaflor-resistant and acetamiprid-resistant strains in two A.gossypii populations with different basal insecticide resistance levels,and evaluated the effects of basal insecticide resistance on the resistance development and cross-resistance,as well as differences in fitness.Under the same selection pressure,Yarkant A.gossypii(with low basal insecticide resistance)evolved resistance to sulfoxaflor and acetamiprid more quickly than Jinghe A.gossypii(with high basal insecticide resistance),and the evolution of A.gossypii resistance to sulfoxaflor developed faster than acetamiprid in both Yarkant and Jinghe,Xingjiang,China.The sulfoxaflor-resistant strains selected from Yarkant and Jinghe developed significant cross-resistance to acetamiprid,imidacloprid,thiamethoxam and pymetrozine;while the acetamiprid-resistant strains developed significant cross-resistance to sulfoxaflor,imidacloprid,thiamethoxam,pymetrozine,and chlorpyrifos.The relative fitness of A.gossypii decreased as the resistance to sulfoxaflor and acetamiprid developed.The relative fitness levels of the sulfoxaflor-resistant strains(Yarkant-SulR and Jinghe-SulR)were lower than those of the acetamipridresistant strains(Yarkant-AceR and Jinghe-AceR).In addition,the relative fitness levels of sulfoxaflor-and acetamiprid-resistant strains were lower in Jinghe than in Yarkant.In summary,basal insecticide resistance of A.gossypii and insecticide type affected the evolution of resistance to insecticides in A.gossypii,as well as cross-resistance to other insecticides.The sulfoxaflor-and acetamiprid-resistant A.gossypii strains had obvious fitness costs.The results of this work will contribute to the insecticide resistance management and integrated management of A.gossypii.

A sublethal concentration of afidopyropen suppresses the population growth of the cotton aphid, Aphis gossypii Glover(Hemiptera: Aphididae)

The cotton aphid,Aphis gossypii Glover(Hemiptera:Aphididae),is an important insect pest of cotton crops worldwide.The objectives of this study were to determine the acute toxicity of afidopyropen and whether there are any effects of sublethal concentrations of afidopyropen on the biological characteristics of A.gossypii.The results showed that afidopyropen possesses high acute toxicity to adult A.gossypii with a 72 h LC_(50) value of 1.062 mg L^(–1).A sublethal concentration(LC_(10))of afidopyropen significantly decreased adult longevity,fecundity and oviposition days of female adults in both F0 and F1 generations.The total pre-adult survival of F_(1) progeny was also significantly reduced by 30%at the LC_(10) of afidopyropen.In addition,the nymph developmental time,pre-adult period,adult pre-reproductive period(APRP),and total pre-reproductive period(TPRP)of the F1 progeny were significantly prolonged compared with the control.Several population parameters,including the net reproductive rate(R_(0)),intrinsic rate of increase(r)and finite rate of increase(λ)of F_(1) progeny were significantly decreased by a sublethal afidopyropen concentration exposure.These results indicated that sublethal concentration of afidopyropen can significantly suppress A.gossypii population growth.It would be useful for assessing the overall effects of afidopyropen on A.gossypii.

The influence of Tetranychus cinnabarinus-induced plant defense responses on Aphis gossypii development

Carmine spider mites （Tetranychus cinnabarinus） and cotton aphids （Aphis gossypii） are both serious pests of cotton, and cause reductions in yields of this key agricultural crop. In order to gain insights into how plant defense responses induced by one herbivore species affect the behavior and performance of another, we examined how infestation with T. cinnabarinus influences the development of A. gossypfi using cotton as a model. In this study, we measured the activities of several important biochemical markers and secondary metabolites in the leaves of cotton seedlings responding to infestation by T. cinnabarinus. Furthermore, the influences of T. cinnabarinus infestation on the development ofA. gossypfi in cotton were also examined. Our data showed that the activities of several key defense enzymes, including phenylalanine ammonia-lyase （PAL）, peroxidase （POD）, lipoxygenase （LOX）, and polyphenol oxidase （PPO）, were substantially increased in cotton seedlings responding to spider mite infestation. Further, the contents of gossypol and condensed tannins, key defensive compounds, were significantly enhanced in leaves of cotton seedlings following T. cinnabarinus infestation. Moreover, the T. cinnabarinus-induced production of defense enzymes and secondary metabolites was correlated with infestation density. The developmental periods of A. gossypii on cotton seedling leaves infested with T. cinnabarinus at densities of 10 and 15 individuals cm-2 were 1.16 and 1.18 times that of control, respectively. Meanwhile, the mean relative growth rates of A. gossypfi on cotton leaves infested with T. cinnabarinus at densities of 8, 10 and 15 individuals cm-2 were significantly reduced. Therefore, these data suggested that the developmental periods of A. gossypfi were significantly lengthened and the mean relative growth rates were markedly reduced when cotton aphids were reared on plants infested with high densities of spider mites. This research sheds light on the role that inducible defense responses played in plant-mediated interspecific interactions between T. cinnabarinus and A. gossypfi.

The damage risk evaluation of Aphis gossypii on wheat by host shift and fitness comparison in wheat and cotton

Aphids are considered as one of the key pests for wheat production worldwide. Major aphid species that infest wheat in China include Sitobion avenae, Rhopalosiphum padi, Schizaphis graminum and Metopolophium dirhodum. However, during our wheat field survey in Wenshang County of Shangdong＇Province, China, we observed that Aphis gossypii can feed on wheat. The damage risk of A. gossypii on wheat was assessed using host shift method. A population of A. gossypii collected from a wheat field in 2015 and another population reared on cotton under laboratory conditions for a decade without exposure to insecticides were used in the study. The results of host shift demonstrated that the A. gossypfi colony from wheat has not yet developed wheat specialization. Moreover, the assessment of A. gossypii fitness on wheat and cotton showed that fecundity and net reproductive rate of A. gossypii population fed on wheat was significantly higher comparing to the population fed on cotton, whether the initial host of A. gossypii population was wheat or cotton. This study raises a warning that the cotton aphid has potential to establish well on wheat and it may cause significant effects under specific circumstances. Therefore, future studies are required to evaluate the effects of A. gossypfi on wheat production.

Comparative transcriptional analysis and identification of hub genes associated with wing differentiation of male in Aphis gossypii

Background:Aphis gossypii Glover(Hemiptera:Aphididae),a worldwide polyphagous phloem-feeding agricultural pest,has three wing morphs(winged parthenogenetic female,gynopara,and male)in the life cycle.The exclusive males could fly from summer hosts to winter hosts,which are essential for gene exchanges of cotton aphid populations from different hosts or regions.However,the molecular mechanism of wing differentiation of male in A.gossypii remains unclear.Results:Morphological observation of male A.gossypii showed that there is no distinct difference in the external morphologies of the 1st and 2nd instar nymphs.The obvious differentiation of wing buds started in the 3rd instar nymph and was visible via naked eyes in the 4th instar nymphal stage,then adult male emerged with full wings.According to morphological dynamic changes,the development of wings in males were divided into four stages:preliminary stage(the 1st instar to 2nd instar),prophase(the 3rd instar),metaphase(the 4th instar),anaphase(the 5th instar).Results of feeding behavior monitoring via EPG(electrical penetration graph)technology indicated that although the male cotton aphids had strong desire to feed(longer duration of C 55.24%,F 5.05%and Pd waves 2.56%),its feeding efficiency to summer host cotton was low(shorter E13.56%and E2 waves 2.63%).Dynamic transcriptome analysis of male aphid at 5 different developmental periods showed that in the 3rd instar nymph,the number of up-regulated DEGs was significant increased,and time-course gene transcriptional pattern analyses results also showed that numerous genes categorized in clusters 3,5,and 8 had the highest expressed levels,which were consistent with morphological changes of wing buds.These results indicate that the 3rd instar nymph is the critical stage of wing bud differentiation in males.Furthermore,through pathway enrichment analysis of DEGs and WGCNA,it revealed that the neuroactive ligand-receptor interaction,Ras signaling pathway,dopaminergic synapse,circadian entrainment and the corresponding hub genes of PLK1,BUB1,SMC2,TUBG,ASPM,the kinesin family members(KIF23,KIF20,KIF18-19)and the novel subfamily of serine/threonine(Aurora kinase A and Aurora kinase B)probably played an important role in the critical stage of wing bud differentiation.Conclusion:This study explored morphological changes and genes transcriptional dynamics males in cotton aphid,revealed the phenomenon of low feeding efficiency of winged males on summer host cotton,and identified key signaling pathways and potential hub genes potentially involved in wing bud differentiation of male in A.gossypii.

Biochemical Characterization of Detoxifying Enzymes in Dimethoate-Resistant Strains of Melon Aphid, Aphis gossypii(Hemiptera: Aphididae)

The melon aphid, Aphis gossypii Glover (Hemiptera: Aphididae), is a highly polyphagous sap sucking pest on wide varieties of crops including cotton and vegetables. It is a notorious vector of many plant viruses that are persistently and non-persistently transmitted. In nature, aphids are regulated by their natural enemies. However, chemical control remains a major management tool even though resistance to insecticides has been documented worldwide. A better understanding of mechanisms by which insecticide resistance occurs and its early detection is desirable to develop effective management strategies. The present investigation was conducted to study the development of resistance to an organophosphate (OP) compound-dimethoate, identify biochemical mechanism(s) involved in resistance and study cross-resistance to imidacloprid in laboratory selected A. gossypii strains in comparison to susceptible strains. Bioassay studies revealed that the LC50 values increased dramatically with dimethoate selection in resistant strains and the resistance ratio (RR) was 270-, 243- and 210-fold greater than that of the susceptible strains by 30th generation. Further, biochemical assays revealed enhanced activities of carboxylesterases (CarE), glutathione S-transferases (GSTs) and cytochrome P450-mediated p-Nitroanisole O-demethylase (PNOD) in resistant strains supporting their role in dimethoate detoxification. This study thus revealed that enhanced activity of detoxifying enzymes viz., CarE, GSTs and PNODs is one of the mechanisms underlying dimethoate resistance in A. gossypii collected from South India. Interestingly, the possibility of negatively correlated cross-resistance to imidacloprid was identified in three OP- resistant strains exhibiting 2.97-, 2.56- and 3.76-fold sensitivity to imidacloprid (a novel neonicotinoid). This indicated that the latter was less affected by the resistance mechanism(s) present.

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.

Differential mRNA expression levels and gene sequences of carboxylesterase in both deltamethrin resistant and susceptible strains of the cotton aphid, Aphis gossypii

Extensive use of insecticides on cotton has prompted resistance development in the cotton aphid, Aphis gossypii （Glover） in China, A deltamethrin-selected population of cotton aphids from Xinjiang Uygur Autonomous Region, China with 228,59-fold higher resistance to deltamethrin was used to examine how carboxylesterase conferred resistance to this pyrethroid insecticide. The carboxylesterase activity in the deltamethrin-resistant strain was 3.67-, 2,02- and 1.16-fold of the susceptible strain when using α-naphthyl acetate （α-NA）, β-naphthyl acetate （β-NA） and α-naphthyl butyrate （α-NB） as substrates, respectively, Carboxylesterase cDNA was cloned and sequenced from both deltamethrinresistant and susceptible strains. The cDNA contained 1581 bp open reading frames （ORFs） coding a 526 amino acid protein. Only one amino acid substitution （Val^87-Ala） was observed between deltamethrin-resistant and susceptible strains but it is not genetically linked to resistance by the catalytic triad and signature motif analysis. The real-time polymerase chain reaction analysis indicated that the resistant strain had a 6.61-fold higher level of carboxylesterase mRNA than the susceptible strain. The results revealed that up-regulation of the carboxylesterase gene, not modified gene structure, may be responsible for the development of resistance in cotton aphids to deltamethrin.

A new active piggyBac-like element in Aphis gossypii

Nine piggyBac-like elements （PLEs） were identified from the cotton aphid Aphis gossypii Glover. All the PLEs shared high sequence similarity with each other. However, eight of the nine PLEs were unlikely to encode functional transposase due to the existence of disruptive mutations within the coding regions. The other one PLE contained major characteristics of members in the piggyBac family, including TTAA target site duplications, inverted terminal repeats （ITRs）, and an open reading frame （ORF） coding for a transposase with a putative DDD domain. This one with an intact transposase ORF was named AgoPLE1.1. The predicted transposase shared 47% similarity with that of Trichoplusia ni piggyBac IFP2. Phylogenetic analyses showed that AgoPLE1.1 was most related to the Heliothis virescens PLE1.1 （HvPLE1.1） element, with 45% and 60% similarity at the nucleotide and amino acid levels, respectively. A functional assay demonstrated that AgoPLE1.1 encoded a functional transposase and was able to cause precise excision in cell cultures. On the other hand, few genomic insertion polymorphisms of AgoPLEI were observed in the genome of the cotton aphid. These observations suggested that AgoPLE1.1 was a PLE that invaded the cotton aphid genome in recent periods and retained its activity.

Sieve element occlusion provides resistance against Aphis gossypii in TGR-1551 melons

Feeding behavior and plant response to feeding were studied for the aphid Aphis gossypii Glover on susceptible and resistant melons(cv.Iroquois and TGR-1551,respectively).Average phloem phase bout duration on TGR-1551 was<7% of the duration on Iroquois.Sixty-seven percent of aphids on TGR-1551 never produced a phloem phase that attained ingestion(EPG waveform E2)in contrast to only 7% of aphids on Iroquois.Average bout duration of waveform E2(scored as zero if phloem phase did not attain E2)on TGR-1551 was<3% of the duration on Iroquois.Conversely,average bout duration of EPG waveform El(sieve element salivation)was 2.8 times greater on TGR-1551 than on Iroquois.In a second experiment,liquid nitrogen was used to rapidly cryofix leaves and aphids within a few minutes after the aphids penetrated a sieve element.Phloem near the penetration site was then examined by confocal laser scanning microscopy.Ninety-six percent of penetrated sieve elements were occluded by protein in TGR-1551 in contrast to only 28% in Iroquois.Usually in TGR-1551,occlusion was also observed in nearby nonpenetrated sieve elements.Next,a calcium channel blocker,trivalent lanthanum,was used to prevent phloem occlusion in TGR-1551,and A.gossypii feeding behavior and the plants phloem response were compared between lanthanum-treated and control TGR-1551.Lanthanum treatment eliminated the sieve element protein occlusion response and the aphids readily ingested phloem sap from treated plants.This study provides strong evidence that phloem occlusion is a mechanism for resistance against A.gossypii in TGR-1551.

Morph-specific differences in metabolism related to flight in the wing-dimorphic Aphis gossypii

The cotton aphid, Aphis gossypii Glover, is a wing-dimorphic species, which causes globally important agricultural losses. In this present study, we compared the biochemical basis of wing polymorphism in A. gossypii with respect to trade-off of energy resources, including glycogen, trehalose, lipids （total lipid, triglyceride and phospholipid）, free fatty acids, and soluble protein between dispersal and reproduction morphs during the wing-bud nymph and adulthood. Total lipid, triglyceride and free fatty acids were significantly higher in winged versus wingless morphs at 12 h of adulthood, the period during which alates are able to fly. By contrast, the wingless morph contained more glycogen than the winged morph from the 4th nymphal stage to adulthood. Trehalose content in the wingless morph was also higher than that in the winged morph during the 3rd and 4th nymphal stages, but vice versa at 12 h of adulthood. Finally, soluble protein content increased from nymphs to adults and was higher during adulthood in aptera versus alate. Whole-body water content in 12-h adults was significantly higher in apterae than that in alatae. These results indicate significant physiological differences between morphs related to specialization for flight.

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.

释放异色瓢虫对甜瓜蚜虫的防控效果

为探究在设施栽培条件下,释放异色瓢虫(Harmonia axyridis)防治甜瓜蚜虫(Aphis gossypii Glover)的综合效果,按不同益害比释放异色瓢虫,评价其在设施条件下对甜瓜蚜虫的防控效果以及天敌和害虫的消长动态。结果表明:异色瓢虫对于甜瓜蚜虫具有良好的防控效果,特别是长期防控效果优于化学农药,益害比1∶5与益害比1∶10的投放密度对蚜虫的防控效果好,两者比较差异未达显著水平,在释放后28 d时防效仍在70%以上。但考虑到生产成本,益害比1∶10为甜瓜上异色瓢虫释放的最佳密度;前期异色瓢虫定殖量随着蚜虫数量增加而升高,14 d时天敌投放处理的瓢虫定殖量达到峰值,故应结合病虫害测报,在蚜虫发生高峰期前尽早释放。

Studies on the Host Biotypes and Its Cause of Cotton Aphidin Nanjing,China

The fitness of cotton aphid, Aphis gossypii Glover, on different host plants was studied. The results showed that the cotton aphid population in Nanjing formed host preference bio-types on cucumber and cotton. The aphids from cotton and cucumber could not inter-transplant successfully between the two host plants. The apterous and alate offspring of stem mother from hibiscus could not survive, reproduce and form population normally when they were transplanted to cucumber and water melon, but they could form normally when transplanted to cotton and muskmelon. There was no significant difference between the stylet length of cotton aphids from cucumber and cotton, so the stylet length was not the cause of the host preference. When the aphids originated on hibiscus were transplanted to cotton, water melon and muskmelon, the stylet could reach host phloem to absorb host plant’s nutrients, but it couldn’t when transplanted to cucumber. Cotton aphid can feed on muskmelon without secretion of water-soluble saliv

内吸性杀虫剂对瓜蚜生物活性及种群实时增长率的影响

为探索新型精准施药方式,筛选防治瓜蚜(Aphis gossypii Glover)的高效内吸性药剂,采用茎部用药法,测定了9种内吸性杀虫剂对黄瓜(Cucumis sativus Linn.)瓜蚜的校正致死率及试验期间的种群实时增长率。结果表明,20%啶虫脒可溶液剂和10%氟啶虫酰胺散粒剂对瓜蚜的内吸活性最高,分别在施用杀虫剂后10、15 d达到100.0%的校正致死率,且施用杀虫剂后20 d校正致死率均稳定在95%以上。各杀虫剂中,10%氟啶虫酰胺散粒剂速效性最差,但持效性最好;其次为70%吡虫啉散粒剂和10%烯啶虫胺水剂,最大校正致死率分别出现在施用杀虫剂后10 d,分别为99.3%和95.0%,且施用杀虫剂后20 d均稳定在90%以上;25%噻虫嗪散粒剂、10%溴氰虫酰胺悬浮剂、25%吡蚜酮可湿性粉剂、20%呋虫胺散粒剂和22.4%螺虫乙酯悬浮剂所有浓度下的最大致死率均低于90%。中、高浓度处理下,20%啶虫脒可溶液剂和70%吡虫啉散粒剂在试验中期对瓜蚜种群增长有明显的阶段性抑制作用,10%氟啶虫酰胺散粒剂在后期持续抑制瓜蚜种群增长。这说明啶虫脒、吡虫啉和氟啶虫酰胺对黄瓜瓜蚜具有较高的内吸活性。

The impact of allelochemicals on the differential expression of symbiotic bacteria in cotton aphids

Insects have developed a good adaptive mechanism in response to environmental stresses in the long-term evolution. They have developed a helpful metabolism system to resist plant allelochemicals. Insects also harbor different kinds of symbiotic bacteria, which provide them a competitive advantage. Here, using cotton aphid as an example, we investigated the effects of four plant allelochemicals on the differential expression of symbiotic bacteria based on transcriptome data. We also studied the composition of symbiotic bacteria and function on pathway level in three kinds of aphids. We found that the bacteria have a significant role in resisting the plant allelochemicals stress and host plant selection by aphids. These results should be useful to investigate the environmental adaption mechanism of aphids in the view of symbiotic bacteria. These results would offer a new insight for improving strategy of aphids and developing new pest control systems.

Stylet penetration behaviors of the cotton aphid Aphis gossypfi 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 Cry 1A （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.

Effects of elevated CO2 and plant genotype on interactions among cotton, aphids and parasitoids

Effects of CO2 level （ambient vs. elevated） on the interactions among three cotton （Gossypium hirsutum） genotypes, the cotton aphid （Aphis gossypii Glover）, and its hymenoptera parasitoid （Lysiphlebiajaponica Ashrnead） were quantified. It was hypothesized that aphid-parasitoid interactions in crop systems may be altered by elevated CO2, and that the degree of change is influenced by plant genotype. The cotton genotypes had high （M9101）, medium （HZ401） and low （ZMS13） gossypol contents, and the response to elevated CO2 was genotype-specific. Elevated C02 increased the ratio of total non-structural carbohydrates to nitrogen （TNC ： N） in the high-gossypol genotype and the mediumgossypol genotype. For all three genotypes, elevated CO2 had no effect on concentrations of gossypol and condensed tannins. A. gossypii fitness declined when aphids were reared on the high-gossypol genotype versus the low-gossypol genotype under elevated CO2. Furthermore, elevated CO2 decreased the developmental time of L. japonica associated with the high-gossypol genotype and the low-gossypol genotype, but did not affect parasitism or emergence rates. Our study suggests that the abundance of A. gossypii on cotton will not be directly affected by increases in atmospheric CO2. We speculate that A. gossypii may diminish in pest status in elevated COz and high-gossypol genotype environments because of reduced fitness to the high-gossypol genotype and shorter developmental time of L. japonica.

Ultrastructure of compatible and incompatible interactions in phloem sieve elements during the stylet penetration by cotton aphids in melon

Resistance of the melon line TGR-1551 to the aphid Aphis gossypii is based on preventing aphids from ingesting phloem sap. In electrical penetration graphs （EPGs）, this resistance has been characterized with A. gossypii showing unusually long phloem salivation periods （waveform El） mostly followed by pathway activities （waveform C） or if followed by phloem ingestion （waveform E2）, ingestion was not sustained for more than 10 min. Stylectomy with aphids on susceptible and resistant plants was performed during EPG recording while the stylet tips were phloem inserted. This was followed by dissection of the penetrated leaf section, plant tissue fixation, resin embedding, and ultrathin sectioning for transmission electron microscopic observation in order to study the resistance mechanism in the TGR. The most obvious aspect appeared to be the coagulation of phloem proteins inside the stylet canals and the punctured sieve elements. Stylets of 5 aphids per genotype were amputated during sieve element （SE） salivation （El） and SE ingestion （E2）. Cross-sections of stylet bundles in susceptible melon plants showed that the contents of the stylet canals were totally clear and also, no coagulated phloem proteins occurred in their punctured sieve elements. In contrast, electron-dense coagulations were found in both locations in the resistant plants. Due to calcium binding, aphid saliva has been hypothesized to play an essential role in preventing/suppressing such coagulations that cause occlusion of sieves plate and in the food canal of the aphid＇s stylets. Doubts about this role of E 1 salivation are discussed on the basis of our results.

Influences of elevated CO2 and pest damage on the allocation of plant defense compounds in Bt-transgenic cotton and enzymatic activity of cotton aphid

Plant allocation to defensive compounds by elevated CO2-grown nontransgenic and transgenic Bt cotton in response to infestation by cotton aphid, Aphis gossypii （Glover） in open-top chambers under elevated CO2 were studied. The results showed that significantly lower foliar nitrogen concentration and Bt toxin protein occurred in transgenic Bt cotton with and without cotton aphid infestation under elevated CO2. However, significantly higher carbon/nitrogen ratio, condensed tannin and gossypol were observed in transgenic Bt cotton ＂GK-12＂ and non-transgenic Bt cotton ＇Simian-3＇ under elevated CO2. The CO2 level and cotton variety significantly influenced the foliar nitrogen, condensed tannin and gossypol concentrations in the plant leaves after feeding by A. gossypii. The interaction between CO2 level x infestation time （24 h, 48 h and 72 h） showed a significant increase in cotton condensed tannin concentrations, while the interaction between CO2 level x cotton variety significantly decreased the true choline esterase （TChE） concentration in the body ofA. gossypi. This study exemplified the complexities of predicting how transgenic and non-transgenic plants will allocate defensive compounds in response to herbivorous insects under differing climatic conditions. Plant defensive compound allocation patterns and aphid enzyme changes observed in this study appear to be broadly applicable across a range of plant and herbivorous insect interactions as CO2 atmosphere rises.