Myzus persicae (green peach aphid) feeding on Arabidopsis induces the formation of a deterrent indole glucosinolate

Cruciferous plants produce a wide variety of glucosinolates as a protection against herbivores and pathogens. However, very little is known about the importance of individual glucosinolates in plant defense and the regulation of their production in response to herbivory. When Myzus persicae (green peach aphid) feeds on Arabidopsis aliphatic glucosinolates pass through the aphid gut intact, but indole glucosinolates are mostly degraded. Although aphid feeding causes an overall decrease in Arabidopsis glucosinolate content, the production of 4-methoxyindol-3-ylmethylglucosinolate is induced. This altered glucosinolate profile is not a systemic plant response, but is limited to the area in which aphids are feeding. Aphid feeding on detached leaves causes a similar change in the glucosinolate profile, demonstrating that glucosinolate transport is not required for the observed changes. Salicylate-mediated signaling has been implicated in other plant responses to aphid feeding. However, analysis of eds5, pad4, npr1 and NahG transgenic Arabidopsis, which are compromised in this pathway, demonstrated that aphid-induced changes in the indole glucosinolate profile were unaffected. The addition of purified indol-3-ylmethylglucosinolate to the petioles of cyp79B2 cyp79B3 mutant leaves, which do not produce indole glucosinolates, showed that this glucosinolate serves as a precursor for the aphid-induced synthesis of 4-methoxyindol-3-ylmethylglucosinolate. In artificial diets, 4-methoxyindol-3-ylmethylglucosinolate is a significantly greater aphid deterrent in the absence of myrosinase than its metabolic precursor indol-3-ylmethylglucosinolate. Together, these results demonstrate that, in response to aphid feeding, Arabidopsis plants convert one indole glucosinolate to another that provides a greater defensive benefit.     

Myzus persicae (green peach aphid) salivary components induce defence responses in Arabidopsis thaliana

Myzus persicae (green peach aphid) feeding on Arabidopsis thaliana induces a defence response, quantified as reduced aphid progeny production, in infested leaves but not in other parts of the plant. Similarly, infiltration of aphid saliva into Arabidopsis leaves causes only a local increase in aphid resistance. Further characterization of the defence-eliciting salivary components indicates that Arabidopsis recognizes a proteinaceous elicitor with a size between 3 and 10 kD. Genetic analysis using well-characterized Arabidopsis mutants shows that saliva-induced resistance against M. persicae is independent of the known defence signalling pathways involving salicylic acid, jasmonate and ethylene. Among 78 Arabidopsis genes that were induced by aphid saliva infiltration, 52 had been identified previously as aphid-induced, but few are responsive to the well-known plant defence signalling molecules salicylic acid and jasmonate. Quantitative PCR analyses confirm expression of saliva-induced genes. In particular, expression of a set of O -methyltransferases, which may be involved in the synthesis of aphid-repellent glucosinolates, was significantly up-regulated by both M. persicae feeding and treatment with aphid saliva. However, this did not correlate with increased production of 4-methoxyindol-3-ylmethylglucosinolate, suggesting that aphid salivary components trigger an Arabidopsis defence response that is independent of this aphid-deterrent glucosinolate.     

Identification of indole glucosinolate breakdown products with antifeedant effects on Myzus persicae (green peach aphid)

The cleavage of glucosinolates by myrosinase to produce toxic breakdown products is a characteristic insect defense of cruciferous plants. Although green peach aphids ( Myzus persicae ) are able to avoid most contact with myrosinase when feeding from the phloem of Arabidopsis thaliana , indole glucosinolates are nevertheless degraded during passage through the insects. A defensive role for indole glucosinolates is suggested by the observation that atr1D mutant plants, which overproduce indole glucosinolates, are more resistant to M. persicae , whereas cyp79B2 cyp79B3 double mutants, which lack indole glucosinolates, succumb to M. persicae more rapidly. Indole glucosinolate breakdown products, including conjugates formed with ascorbate, glutathione and amino acids, are elevated in the honeydew of M. persicae feeding from atr1D mutant plants, but are absent when the aphids are feeding on cyp79B2 cyp79B3 double mutants. M. persicae feeding from wild-type plants and myrosinase-deficient tgg1 tgg2 double mutants excrete a similar profile of indole glucosinolate-derived metabolites, indicating that the breakdown is independent of these foliar myrosinases. Artificial diet experiments show that the reaction of indole-3-carbinol, a breakdown product of indol-3-ylmethylglucosinolate, with ascorbate, glutathione and cysteine produces diindolylmethylcysteines and other conjugates that have antifeedant effects on M. persicae . Therefore, the post-ingestive breakdown of indole glucosinolates provides a defense against herbivores such as aphids that can avoid glucosinolate activation by plant myrosinases.     

Redox responses of Arabidopsis thaliana to the green peach aphid,Myzus persicae

The green peach aphid (Myzus persicae) is a phloem-feeding insect that causes economic damage on a wide array of crops. Using a luminol-based assay, a superoxide-responsive reporter gene (Zat12::luciferase), and a probe specific to hydrogen peroxide (HyPer), we demonstrated that this aphid induces accumulation of reactive oxygen species (ROS) in Arabidopsis thaliana. Similar to the apoplastic oxidative burst induced by pathogens, this response to aphids was rapid and transient, with two peaks occurring within 1 and 4 hr after infestation. Aphid infestation also induced an oxidative response in the cytosol and peroxisomes, as measured using a redox-sensitive variant of green fluorescent protein (roGFP2). This intracellular response began within minutes of infestation but persisted 20 hr or more after inoculation, and the response of the peroxisomes appeared stronger than the response in the cytosol. Our results suggest that the oxidative response to aphids involves both apoplastic and intracellular sources of ROS, including ROS generation in the peroxisomes, and these different sources of ROS may potentially differ in their impacts on host suitability for aphids.     

The Arabidopsis thaliana/Myzus persicae model system demonstrates that a single gene can influence the interaction between a plant and a sap-feeding insect

We have developed an Arabidopsis thaliana/Myzus persicae model system to allow the dissection of plant/insect interactions at a molecular genetic level. This allows the examination of the role of single plant genes in the interaction between the plant and an aphid. Our initial studies have exploited an Arabidopsis genotype in which the function of the amino acid transporter ANT1 has been abolished. This mutation results in a change in the proportions of several amino acids within the phloem sieve elements (SEs) resulting in an increase in the proportion of essential amino acids. This has been measured using aphid stylectomy to collect SE samples, followed by a novel micellar electrokinetic chromatography method for amino acid analysis. The SE content represents the aphid's diet, and use of electrical penetration graph technology and honeydew clocks have demonstrated that this altered diet results in a change in the feeding rate of the aphid. Balance sheets can be produced to show the amount (nmoles/24 h) of each of 18 amino acids taken up and excreted by aphids feeding on wild type and ant1 mutant plants. The data show that aphids feeding on the ant1 mutant take up larger amounts of amino acids. However, we could not detect any effect on the reproductive rate of the aphids. The results show that, under experimental conditions, this model system can be used to identify plant genes that control the behaviour and fecundity of an insect pest.     

球孢白僵菌对温室蚜虫的侵染生物学

研究了球孢白僵菌在不同浓度、施菌方式、温度和相对湿度下对温室蚜虫的侵染力以及接种后对蚜虫的侵染速率。结果表明,高(109个孢子.mL-1)、中(108个孢子.mL-1)、低(107个孢子.mL-1)3个浓度剂量对蚜虫都有较强的致病力,且浓度越大,蚜虫的死亡率越高,死亡时间越提前。用浸渍法和孢子浴法接种蚜虫,6 d的累积死亡率分别为100%和31.1%。在测试的3个温度(22、26、30℃)中,26℃时蚜虫侵染力最强,第3天出现死亡高峰,第5天时累积死亡率就达到100%,明显高于22、30℃的处理。相对湿度越大,球孢白僵菌的致病力越强,蚜虫死亡速度越快。在温湿度组合中相对湿度为95%时,温度对白僵菌的侵染力几乎无影响,但影响发病速度,相对湿度低于95%时,26℃的侵染力始终高于22和30℃时的侵染力。通过接种后不同时间段用0.2%百菌清处理蚜虫测定该菌株的侵染速率,结果表明接种后24 h是该球孢白僵菌有效侵染蚜虫的关键时期。     

春季烟蚜有翅蚜空间分布及迁入量估计的初步研究

本文通过对春季烟田烟蚜有翅蚜的抽样调查,研究有翅蚜的空间分布并进行迁人量估计。采用频次分布方法进行有翅蚜空间分布型测定,拟合结果既符合泊松分布又符合负二项分布;通过计算有翅蚜在不同叶位上的概率分布,结果表明,有翅蚓适入后大部分集中于烟株上部;通过定期调查烟株有翅蚜数量,并采用定积分中的抛物线法近似计算有翅蚜迁入总量,结果表明,有翅蚜迁入量约为４７９头／百株。     

云南主要烟区烟蚜抗药性的监测

采用浸叶法测定了云南省红河弥勒、楚雄东华、昆明寻甸、丽江七河及昭通昭阳共5个烟区的烟蚜Myzuspersicae(Sulzer)种群对7种常用杀虫剂的抗性水平。结果表明,5个烟区烟蚜对甲胺磷均产生了中等或中等程度以上抗性。红河弥勒和昭通昭阳两烟区烟蚜对氧化乐果产生了中等或中等程度以上抗性;楚雄东华、红河弥勒、昭通昭阳和丽江七河烟区烟蚜对硫丹产生了中等或中等程度以上抗性。对其余杀虫剂尚处于低抗以下水平。     

桃蚜MpAChE基因RNAi表达载体构建及转化

通过害虫取食植物表达害虫发育关键基因dsRNA的转基因植株,分析能否通过抑制害虫特定基因的表达来防控害虫。本研究利用RT-PCR技术从桃蚜中克隆乙酰胆碱酯酶基因383 bp cDNA片段,命名为MpAChE。进一步利用该MpAChE基因片段构建植物RNAi表达载体RNAi-MpAChE,并通过浸花法转化野生型拟南芥,通过卡那霉素抗性筛选转化植株,PCR及Southern杂交进一步鉴定转基因植株。结果表明:克隆的cDNA片段与桃蚜中已克隆的乙酰胆碱酯酶(GenBank登录号AY147797)cDNA序列核苷酸一致性为99%。卡那霉素抗性初步筛选和PCR进一步鉴定,获得25株阳性转基因植株。从25株中随机选择的5株阳性植株,Southern杂交均为阳性。经接种桃蚜初步鉴定,转基因植株对蚜虫的抗性效果不显著。     

四株蜡蚧轮枝菌对桃蚜的侵染力及对其生殖力的影响

通过研究来自不同寄主的 4株蜡蚧轮枝菌菌株对桃蚜的侵染致死力、发病强度指数、以及对无翅成蚜生殖力及子代的影响 ,综合探讨病菌的侵染致病力及其潜力。试验 (2 4℃ ,10 0 % RH,5× 10 6孢子 /ml)结果表明 ,蜡蚧轮枝菌对无翅成蚜的侵染致死力和发病指数总体上是一致的 ,但二者在 4菌株间有明显差异 ,强弱依次为 VL FNL 95≥ VLNTV94 >VL ASA87>VLKTV79;对不同虫态 ,二者也有较大差异 ,对无翅成蚜明显高于有翅若蚜和 2～ 3龄若蚜。上述 4菌株对无翅成蚜的感染死亡高峰 ,依次为 4～ 5天、4～ 7天、4～ 8天和 5～ 9天 ,感染死亡分别为 84 .2 1～ 10 0 %、75.0 0～ 10 0 %、4 8.2 8～ 10 0 %和 2 9.6 3～ 77.78%。其中对 2～ 3龄若蚜 ,菌株间差异不很大。前 2个菌株 6天可达最高发病指数 ,虫体表面布满菌丝和分生孢子。与对照相比 ,可明显抑制无翅成蚜的生殖力。处理后 3天 ,产仔蚜量明显减少 ;4～ 5天锐减 ;8天种群产仔蚜量降低 39.59～ 4 8.37%。     

烟蚜在山东烟田的发生与消长

对山东烟区烟蚜Myzus persicae(Sulzer)的发生消长规律进行7年的系统调查。结果表明:烟蚜的发生为单峰型曲线,发生高峰大致在每年的5月底6月初,大多数年份不能形成第2个蚜量高峰,且烟蚜的发生以无翅蚜为主。并对以无翅蚜为主以及近年来烟蚜田间发生的数量较少的原因进行分析。     

Host selection by winged colonisers within the Myzus persicae group: a contribution towards understanding host specialisation

Abstract.  1.  Myzus persicae sensu lato demonstrates considerable genetic variation in respect to adaptation to host plants. The subspecies M. persicae nicotianae shows a preference for tobacco, while M. persicae sensu stricto ( s. str. ) for other herbaceous plants. Given that winged colonisers of several aphid species play an important role in selecting host plants, here their role in the host specialisation observed in M. persicae was examined in choice and no-choice tests conducted outdoors, in performance studies, and in DC Electrical Penetration Graph (DC-EPG) studies.
2. In outdoor choice tests, 77% of spring migrants of M. persicae nicotianae chose tobacco, whereas equal proportions of M. persicae s. str. selected tobacco and pepper. In no-choice tests, spring migrants settled more quickly after alighting on host rather than on non host plants, and significantly more alate M. persicae s. str. (27%) than M. persicae nicotianae (2%) left tobacco after walking briefly on the leaf surface, whilst no significant differences were found on pepper. Cross-host transfers significantly reduced the fecundity of both summer and spring migrants of the two subspecies. Finally, the results of no-choice tests and DC-EPG studies showed that winged aphids distinguished their host through cues located on the plant surface or in subcutaneous tissues perceived prior to the initiation of feeding.
3. This study demonstrates the important role of winged colonisers in the evolution of host specialisation in M. persicae . The multifarious divergent selection that the two host forms experience, i.e. the selection against cross-migrants and their subsequent generations, is a crucial factor involved in the development and maintenance of host specialisation and promotes the parallel evolution of improved host-recognition ability.     

桃蚜虫霉病始发期的初始感染与接触传染

自2002年10月至2005年5月在杭州上空诱获桃蚜(Myzus persicae)的迁飞性有翅蚜2351头,将其带回室内在(21±1)℃和12L∶12D条件下单头饲养12d,其中639头在定殖后7d内发病死亡,初始感染率达27·2%。在病死有翅蚜中,99·4%系5种蚜科专化性虫霉侵染所致,新蚜虫疠霉(Pandora neoaphidis)的发生比例高达81·4%。带病迁飞的有翅蚜在发病死亡前具有一定生殖力,定殖后第6天平均累计产若蚜(2·4±0·13)头,远低于同期未带病有翅蚜产下的(11·6±0·33)头。在带病有翅蚜建立的后代蚜群中,接触传染在母蚜死亡2d后即可见到。在定殖后第12天,二级感染占总观察蚜群数的13·3%,三级感染占总观察蚜群数的4·4%,占二级感染蚜群数的33·3%。此时,未带病有翅蚜的后代蚜群的平均活蚜数达(50·6±2·30)头,而带病有翅蚜的后代蚜群平均仅有(21·5±1·98)头。结果表明,虫霉病能够通过寄主带病迁飞传至寄主迁入地,并在后代蚜群中相互传染而起到调节蚜虫数量增长的作用。     

桃蚜对吡虫啉次亚致死用量反应的时间-剂量-死亡率模拟分析

采用电动弥雾法测定了桃蚜对吡虫啉次亚致死浓度(0.01～1.0μg·ml^-1)为期1周的反应。所获数据经时间-剂量-死亡率模拟分析,获得了根据不同期望杀蚜水平的致死浓度和致死时间,且二者互为函数关系,即达到同一期望杀蚜水平的时间越长,所需药液浓度越低,反之亦然.以弥雾后第1d和第5d为例,LC50及其95％置信限分别为2.8(1.8～4.6)和0.5(0．4～0．6)μg·ml^-1,LC90分别为7．1(3．8～13．0)和1.2(1.0～1．5)μg·ml^-1,LC99分别为11.9(6.0～23.9)和2.0(1.5～2．8)μg·ml^-1,当药液浓度为1．0、2．0和3.0μg·ml^-1时,LT50分别为2.9、1.4和1.0d,L1、90分别为5．8、3.6和2．4d。结果表明,吡虫啉的田间常规使用浓度(～33μg·ml^-1)是按快速击倒要求确定的,它比24h内LC99的上限还高;稍放宽时间要求,达到特定杀蚜水平所需的药液浓度可大幅降低。     

Screening for resistance against Myzus persicae and Macrosiphum euphorbiae among wild Solanum

As a consequence of selection for productive traits, the genetic diversity of Solanum tuberosum cultivars has been drastically reduced. With the goal to develop aphid-resistant potato cultivars, our objective was to quantify the resistance of 14 accessions belonging to five wild potato species ( Solanum chomatophilum , Solanum stoloniferum stoloniferum , Solanum bukasovii , Solanum marinasense and Solanum medians ) against two aphid pests, Myzus persicae and Macrosiphum euphorbiae . Aphids were reared under controlled conditions in microcages clipped on the abaxial face of mature leaves of the Solanum species. The five wild Solanum species induced more than 90% of nymph mortality in M. persicae . The mortality rate of Ma. euphorbiae was also >90% on all S. chomatophilum , S. stoloniferum and S. medians accessions and on S. bukasovii -PI 414155. When enough adult aphids emerged, that is for Ma. euphorbiae on three S. bukasovii and three S. marinasense accessions, the prereproductive period, the adult survival and fecundity were assessed. These parameters allowed us to calculate the intrinsic rate of natural increase ( r_m ), the finite rate of increase and the population doubling time (DT). The r_m and the finite rate of increase were significantly reduced, whereas the population's DT was significantly enhanced for most (five out of six) wild Solanum accessions. All accessions of S. chomatophilum , S. stoloniferum and S. medians and S. bukasovii -PI 414155 were highly resistant to both M. persicae and Ma. euphorbiae . In conclusion, S. chomatophilum -PI 310943 and -PI 310990, and S. stoloniferum -PI 195167, -PI 201855 and -PI 275248, can be amenable for potato breeding programmes.     

Spatial and temporal dynamics of Myzus persicae clones in fields and suction traps

1 The population of peach‐potato aphid Myzus persicae in Scotland is comprised almost entirely of long‐term asexual clones. 2 Over a ten year period, M. persicae from Scottish fields and suction traps were analysed with six microsatellite markers. 3 Out of 1497 individuals analysed, 14 clones (denoted A–N) comprised over 98% of the collection. 4 Some clones were particularly abundant but most clones had a widespread distribution on all available plants. 5 Clones E and L had distinct features in their distributions as clone L was geographically totally restricted to the north east of Scotland and clone E showed a marked preference for brassica crops. 6 Clones E and L provide direct evidence of a role for local adaptations in the distribution of M. persicae clones.     

Behavioural responses of the seven‐spot ladybird Coccinella septempunctata to plant headspace chemicals collected from four crop Brassicas and Arabidopsis thaliana,infested with Myzus persicae

1 Insects using olfactory stimuli to forage for prey/hosts are proposed to encounter a ‘reliability–detectability problem’, where the usability of a stimulus depends on its reliability as an indicator of herbivore presence and its detectability. 2 We investigated this theory using the responses of female seven‐spot ladybirds Coccinella septempunctata (Coleoptera: Coccinellidae) to plant headspace chemicals collected from the peach‐potato aphid Myzus persicae and four commercially available Brassica cultivars; Brassica rapa L. cultivar ‘turnip purple top’, Brassica juncea L. cultivar ‘red giant mustard’, Brassica napus L. cultivar ‘Apex’, Brassica napus L. cultivar ‘Courage’ and Arabidopsis thaliana. For each cultivar/species, responses to plants that were undamaged, previously infested by M. persicae and infested with M. persicae, were investigated using dual‐choice Petri dish bioassays and circular arenas. 3 There was no evidence that ladybirds responded to headspace chemicals from aphids alone. Ladybirds significantly preferred headspace chemicals from B. napus cv. Apex that were undamaged compared with those from plants infested with aphids. For the other four species/cultivars, there was a consistent trend of the predators being recorded more often in the half of the Petri dish containing plant headspace chemicals from previously damaged and infested plants compared with those from undamaged ones. Furthermore, the mean distance ladybirds walked to reach aphid‐infested A. thaliana was significantly shorter than to reach undamaged plants. These results suggest that aphid‐induced plant chemicals could act as an arrestment or possibly an attractant stimulus to C. septempunctata. However, it is also possible that C. septempunctata could have been responding to aphid products, such as honeydew, transferred to the previously damaged and infested plants. 4 The results provide evidence to support the ‘reliability–detectability’ theory and suggest that the effectiveness of C. septempunctata as a natural enemy of aphids may be strongly affected by which species and cultivar of Brassica are being grown.     

不同品种烤烟对烟蚜羧酸酯酶和谷胱甘肽转移酶活性的影响

为明确不同烤烟品种对烟蚜Myzus persicae(Sulzer)解毒酶活力的影响,比较了9个品种烤烟上的绿色型烟蚜的解毒酶活力,发现烤烟品种对烟蚜α-乙酸萘酯羧酸酯酶、β-乙酸萘酯羧酸酯酶、谷胱甘肽转移酶活力有显著影响,不同品种烤烟上烟蚜的α-乙酸萘酯羧酸酯酶、β-乙酸萘酯羧酸酯酶、谷胱甘肽转移酶均有差异。烤烟品种V2、云烟87和红大上的烟蚜羧酸酯酶活力较高,云烟317、云烟85和K326居中,K346、K358和G28则较低。9个品种烤烟对烟蚜谷胱甘肽转移酶活力有所影响,但差异不大。     

异色瓢虫对设施栽培桃树桃蚜的捕食功能反应研究

为明确异色瓢虫对设施栽培桃树上桃蚜的自然控制力,在室内研究了异色瓢虫成虫自身密度和不同蚜虫密度对捕食功能的影响。结果表明,异色瓢虫对桃蚜的捕食功能反应符合Holling-Ⅱ圆盘方程,其拟合模型为Na=0.898Nt/(1+0.0045Nt),每头异色瓢虫在1 d内对桃蚜的最大捕食量为200头,捕食每头桃蚜的处置时间Th=0.005d。异色瓢虫自身密度对桃蚜捕食作用有一定制约,拟合Watt竞争模型方程为A=86.441P-0.6592。     

Transmission of potato leafroll virus in relation to the honeydew excretion of Myzus persicae

Honeydew excretion of single Myzus persicae nymphs on potato leafroll virus (PLVR)-infected Physalis floridana was studied during the acquisition access period (AAP) in relation to the efficiency of virus transmission.
With increasing length of the AAP, the percentage of nymphs that transmitted the virus increased. These nymphs produced significantly more honeydew droplets during the AAP on PLRV-infected P. floridana plants than nymphs which failed to transmit the virus. However, the number of honeydew droplets excreted during the AAP by transmitting nymphs did not affect the length of the latency period. Nymphs which infected the first test plant after a short latency period produced a similar amount of honeydew during the AAP to those with a longer latency period.
Honeydew excretion recorded on plants of varied age, showed that nymphs feeding on bottom leaves of infected plants produced more honeydew droplets than on comparable leaves of healthy plants. On infected plants, nymphs produced more honeydew droplets on bottom leaves with pronounced symptoms than on top leaves that hardly showed any symptom of PLRV infection.
The concentration of viral antigen measured by ELISA was lower in top leaves than in bottom leaves of infected plants. Nevertheless, nymphs feeding on top leaves transmitted the virus more efficiently than those which used bottom leaves as virus source. When bottom leaves were used as a virus source, the percentage of viruliferous nymphs decreased with plant age. These results indicate that the availability of virus for acquisition by aphids declines with increasing plant age and symptom severity.