烟粉虱复合种内共生菌多样性及其生物学意义

烟粉虱复合种Bemisia tabaci（Gennadius） complex内共生菌的种类可分为5类：变形菌门（Proteobacteria）γ变形菌纲的初级内共生菌Candidatus Portiera aleyrodidarum和次级内共生菌,变形菌门α变形菌纲Wolbachia属细菌,衣原体门（Chlamydiae）衣原体纲细菌CandidatusFritschea bemisiae以及拟杆菌门（Bacteroidetes）类噬胞细菌（Cytophaga-like-organism, CLO）。系统发育分析表明,烟粉虱初级内共生菌分为2支,一些初级内共生菌和宿主系统发育并不严格一致; 次级内共生菌分为3支,一些次级内共生菌可能存在水平传播或多次侵入的现象。上述5类内共生菌在烟粉虱不同生物型/地理种群内的分布存在差异。B型烟粉虱可能具有独特的初级、次级内共生菌,并且不含有或很少含有Wolbachia属细菌、衣原体纲细菌Candidatus Fritschea bemisiae以及类噬胞细菌（CLO）。最后探讨了不同内共生菌对烟粉虱宿主的生物学意义及其研究方向。     

不同食物对烟粉虱三种寄生蜂取食寄生等行为的影响

【目的】就不同食物对烟粉虱Bemisia tabaci(Gennadius)寄生蜂取食、寄生等行为的影响进行分析,以阐明食物或营养来源对蚜小蜂取食、寄生等生物学行为的影响机制。【方法】以古桥桨角蚜小蜂Eretmocerus furuhashii、丽蚜小蜂Encarsia formosa和双斑恩蚜小蜂Encarsia bimaculata为研究对象,分别以蜂蜜水、2~3龄烟粉虱若虫及清水进行饲喂,观察和研究分析3种食物来源对3种蚜小蜂取食与寄生等行为的影响。【结果】3种蚜小蜂均以取食15%蜂蜜水的蚜小蜂的取食、搜索与寄生等行为频次最高、分配时间最长,而取食清水的蚜小蜂取食、搜索与寄生等行为频次最低、分配时间最短;取食不同食物后,3种蚜小蜂各种行为的变化趋势一致。【结论】不同食物或营养来源对蚜小蜂的取食及寄生等行为有着显著的影响,进而可以影响到蚜小蜂对烟粉虱的生物防治效率。     

Influence of foliar and systemically applied azadirachtin on host‐plant evaluation behaviour of the sweetpotato whitefly,Bemisia tabaci

Abstract The behaviour of female adult Bemisia tabaci is observed for a period of 20 min after initial contact with untreated cucumber leaves, or leaves either foliar or systemically treated with azadirachtin, to determine whether application of azadirachtin affects the host‐evaluation behaviour and whether the behaviour on treated leaves differs between application methods. Application of azadirachtin deters settling of the whiteflies on host plants. The whiteflies probe for shorter duration and less frequently but spend longer and engage more frequently in labial grooming on treated leaves than on untreated leaves. Behavioural transition between probing and other behavioural elements is less common, and that between labial grooming and other behavioural elements more common, on both treated leaves than on untreated leaves. No difference is detected in the host‐evaluation behaviour of B. tabaci between leaves foliar treated and systemically treated with azadirachtin.     

三种抗生素对B型和Q型烟粉虱内共生菌的去除效果比较研究

利用烟粉虱Bemisia tabaci(Gennadius)内共生菌特异性引物,研究了内共生菌在B、Q型烟粉虱种群中的分布和感染率,同时评价了3种不同的抗生素利福平、氨苄青霉素和硫酸卡那霉素分别在3种不同的浓度下(100.0、50.0及25.0μg/mL)对烟粉虱内共生菌的去除效果。结果表明:B、Q型烟粉虱原生内共生细菌Portiera的带菌率均为100.0%;B、Q型烟粉虱次生内共生菌Hamiltonella的带菌率分别为91.7%和100.0%;B型烟粉虱次生内共生菌Rickettsia的带菌率为87.5%,Q型为0;其它次生内共生菌在B、Q型烟粉虱中均未检测到。利福平、氨苄青霉素和硫酸卡那霉素在3种不同的浓度下均不能去除B、Q型烟粉虱Portiera;利福平、氨苄青霉素在3种不同的浓度下均能完全去除B型烟粉虱Rickettsia,硫酸卡那霉素在不同浓度下去除Rickettsia的效果不同;3种抗生素去除Hamiltonella的能力受抗生素种类以及浓度的影响。同一抗生素在不同浓度下去除Hamiltonella的效果均是100.0μg/mL>50.0μg/mL>25.0μg/mL;不同浓度的抗生素去除Hamiltonella的效果均是利福平>氨苄青霉素>硫酸卡那霉素,各浓度与各抗生素之间的去除Hamiltonella的效果均具有显著性差异。     

B型烟粉虱在甘蓝、西葫芦和辣椒上的刺吸取食行为

利用刺吸电波技术（EPG）记录了B型烟粉虱Bemisia tabaci在甘蓝Brassica oleracea L. var. caqitata、西葫芦Cucurbita pepo L.和辣椒Capsicum annuum L.上的刺吸取食波形。结果表明,3种寄主植物的适合度为甘蓝>西葫芦>辣椒。在所观察的25头烟粉虱中,到达甘蓝、西葫芦和辣椒韧皮部的个体数分别为24、19和15,其中持续吸食头数为22、10和1。在第一阶段（从刺吸开始到第一个E(pd)1出现）,烟粉虱在甘蓝、西葫芦和辣椒3种植物上非刺探时间所占比例分别为9.59%、23.55%和36.97%,三者差异显著; 在第二阶段（从第一个E(pd)1出现到第一次韧皮部持续吸食）,烟粉虱在甘蓝和西葫芦上所考察的4个指标无显著差异; 在第三阶段（从第一次韧皮部持续吸食至记录结束）,烟粉虱在甘蓝上第一次韧皮部持续吸食后的刺探次数（4.73）显著低于在西葫芦上的刺探次数（13.40）。在自由状态下,烟粉虱60 min内在甘蓝、西葫芦和辣椒上留存粉虱的百分比分别为100％、88％和28％,这进一步验证了上述依据EPG记录得到的结果。     

The effects of physiological factors and host plant experience on the ovipositional activity of the sweet potato whitefly, Bemisia tabaci

Both egg load and levels of vitellogenin and vitellin increased when Bemisia tabaci Gennadius (Homoptera: Aleyrodidae), were deprived of host plants. These increases resulted in significant increases in rates of oviposition. There was no difference in total fecundity between whiteflies temporarily deprived of hosts and a control treatment that had continuous access to hosts. The results of previous exposure to a preferred host, melon (Cucumis melo), and a non-preferred host, cotton (Gossypium hirsutum), were also examined. B. tabaci deposited significantly more eggs during a 6-h period on melon than on cotton, regardless of host species they were previously confined to. In addition, whiteflies previously exposed to cotton, deposited more eggs than females previously exposed to melon. This higher rate of oviposition resulted in a low egg load as well as lower levels of vitellogenin and vitellin. Conversely, a low rate of oviposition led to high egg load and high levels of vitellogenin and vitellin. Individuals from the same treatment exhibited variation in egg load, which was negatively and significantly correlated with previous oviposition activity. We conclude that the effects of previous host plant exposure on oviposition were, in part, due to behavioral differences that are not related to the reproductive physiology.     

华东、华南和西南地区烟粉虱的种群及寄主分化

烟粉虱Bemisia tabaci是一个复合种,它具有的生物型分化、较强的传播病毒的能力和抗药性、较快的繁殖速率等特征使其成为我国农业生产中重要害虫之一。本研究利用细胞色素线粒体氧化酶Ⅰ基因,对采集自江苏、广东和云南三省的烟粉虱样本进行了生物型鉴定,并对烟粉虱生物型与寄主植物之间的关联性开展了调查。结果表明,在广东和云南省,都存在未鉴定的土著种群与入侵的B型、Q型共存的现象;同时,在本研究中广东省尚未采集到Q型烟粉虱,而在江苏采集到的粉虱样本全部为入侵型。研究结果还表明,相对于入侵种而言,土著种群显示出更强的寄主植物趋同性;丰富的寄主植物以及本身具有的多食性特性有助于B型、Q型等生物型在世界各地的广泛入侵。     

烟粉虱及海氏桨角蚜小蜂对中国番茄黄化曲叶病毒感染烟草的嗅觉反应

双生病毒可通过调控寄主植物促进媒介昆虫烟粉虱种群增长,然而病毒侵染植物后是否通过调控植物挥发物来影响烟粉虱及其天敌的嗅觉反应还未见报道。【目的】本文旨在研究烟草植株感染中国番茄黄化曲叶病毒(Tomato yellow leaf curl China virus,TYLCCNV)后对烟粉虱Bemisia tabaci(Gennadius)及其重要寄生性天敌海氏桨角蚜小蜂Eretmocerus hayati(Zolnerowich and Rose)嗅觉反应行为的影响。【方法】利用Y形嗅觉仪方法,我们测试了烟粉虱及海氏桨角蚜小蜂对带毒植株、健康植株及烟粉虱危害植株的选择偏好性。【结果】烟粉虱及海氏桨角蚜小蜂选择携带TYLCCNV病毒的烟草显著多于健康烟草植株,但烟草被病毒与烟粉虱共同侵染时,烟粉虱对带毒烟草的选择仍显著多于无毒植株,而寄生蜂虽然仍较多选择带毒植株,但无显著差异。【结论】这些结果表明烟粉虱及海氏桨角蚜小蜂偏好选择携带TYLCCNV病毒的烟草,但这种偏好作用在烟粉虱取食共同危害时有一定程度的减弱。本研究首次报道了双生病毒侵染植物可增加烟粉虱及其天敌对植物的选择作用,并就其功能及机制进行了讨论。     

两种棉花防御反应次生代谢物质及其对烟粉虱羧酸酯酶活性的影响

通过烟粉虱Bemisia tabaci(Gennadius)取食为害诱导棉花防御反应次生代谢物质槲皮素和蛋白酶抑制剂的作用,以及2种次生物质对烟粉虱主要解毒酶羧酸酯酶的诱导表达作用,初步探索棉花防御与害虫反防御策略。结果表明烟粉虱取食对棉叶内槲皮素和蛋白酶抑制剂的诱导表达迅速但短暂,取食后1～3d,对槲皮素的最大诱导表达量为2.09mg/g,对蛋白酶抑制剂的最大诱导表达量为1.85U。同时槲皮素和蛋白酶抑制剂2种植物次生代谢物质在0.005%至1%浓度范围内,均能够诱导烟粉虱体内羧酸酯酶比活力过量表达。在浓度为0.1%时,羧酸酯酶的比活力达到最大值,分别为0.0725OD/30min和0.07825OD/30min;2个处理的剂量效应趋势非常相似。     

Delayed development of the whitefly (Bemisia tabaci) and increased parasitism by Encarsia bimaculata in response to sublethal doses of piperonyl butoxide

Abstract Effects of sublethal piperonyl butoxide (PB) on parasitization of Bemisia tabaci (Gennadius ) (Hemiptera: Aleyrodidae) by Encarsia bimaculata Heraty et Polaszek (Hymenoptera: Aphelinidae) were evaluated both in cage and greenhouse experiments. When first, second and third instar B. tabaci nymphs were treated with PB, all but the first instar were significantly prolonged. Data indicated that sublethal PB could improve E. bimaculata parasitism rates without influencing parasitoid eclosion rates. Prolonged development increased rates of parasitism by E. bimaculata, from 17.6% to 24.7% in cages, presumably by increasing the duration of host exposure. Sublethal PB combined with E. bimaculata as an integrated approach to control B. tabaci was evaluated using life table parameters under greenhouse conditions. Indices of population trend (I) calculated from life tables were estimated at 4.6 for B. tabaci exposed to PB and parasitoids compared to 14.1 with parasitoids alone and 23.5 in untreated controls. The results showed that after PB was sprayed and parasitoids introduced, development of B. tabaci was delayed and the peak of each stage was postponed. The older nymphal stage had highest mortality, primarily due to mortality caused by parasitism by E. bimaculata.     

Effect of planting date and nitrogen fertilization on photosynthesis and soluble carbohydrate contents of cotton in relation to silverleaf whitefly （Bemisia tabaci biotype “B”） populations

The impacts of planting date and nitrogen fertilization on cotton （Gossypium hirsutum L.） photosynthesis and soluble carbohydrate contents in relation to silverleaf whitefly, Bemisia tabaci （Gennadius） biotype “B”, populations were examined in field experiments. Cotton planted in late April and early June was treated with 0, 112, 168 and 224 kg/N hectare in soil using urea fertilizer. The mean photosynthetic rate of April-planted cotton was 4%-20% higher than that of June-planted cotton early in the season, but 10%- 18% lower than that of June-planted cotton late in the season. The photosynthetic rates for both planting dates were positively correlated with levels of added nitrogen. While levels of glucose for both planting dates were positively correlated with nitrogen levels, fructose and sucrose levels were not. The mean levels of fructose were up to 40% lower, while that of sucrose were up to 59% higher, in April-planted cotton than in June-planted cotton. Levels of photosynthetic rate or stomatal conductance were not correlated with adult whitefly densities for either planting date. Levels of glucose and fructose were positively correlated with whitefly densities only for June-planted cotton late in the season.     

Effects of temperature on the growth and reproduction characteristics of Bemisia tabaci B‐biotype and Trialeurodes vaporariorum

The development period, survival rate, longevity and fecundity of two whiteflies, Bemisia tabaci B‐biotype and Trialeurodes vaporariorum (Homoptera: Aleyrodidae) were compared under different temperature laboratory conditions (15°C, 18°C, 21°C and 24°C). Egg development of B. tabaci B‐biotype was significantly longer compared with that of T. vaporariorum at 15°C, 18°C and 24°C. Significantly longer pseudo‐pupae development and lower survival rate were found in B. tabaci B‐biotype at 15°C compared with those at 18°C, 21°C and 24°C. Significantly higher fecundity was found in B. tabaci B‐biotype at 24°C compared with that at 15°C, 18°C and 21°C. However, the fecundity of T. vaporariorum was significantly lower at 24°C relative to that at 15°C, 18°C and 21°C. Significantly shorter 1st instar larval development was found in T. vaporariorum compared with that of B. tabaci at 15°C and 18°C. Significantly longer 2nd instar larval development was found in B. tabaci and T. vaporariorum at 15°C compared with that at 18°C, 21°C and 24°C. However, significantly shorter 3rd instar larval development was found in T. vaporariorum compared with that of B. tabaci at 15°C, 18°C and 24°C. The adaptive divergence of tolerance to relatively low temperature may be an important factor that results in the interspecific differentiation between the seasonal dynamics of these two whiteflies in China.     

B型烟粉虱与浙江非B型烟粉虱的竞争

为了了解近年来入侵中国的B型烟粉虱(Bemisiatabaci)取代本地非B型烟粉虱的潜能,在室内将一个B型与一个浙江非B型烟粉虱种群混合饲养在不同寄主植物上,跟踪观察混合种群中两个生物型个体数量相对比例的变化。结果表明,当两种生物型在棉花(Gossypiumhirsutum)上以相同初始数量共存竞争时,经过6代,非B型完全被B型替代;而在西葫芦(Cucurbitapepo)上以相同初始数量共存竞争时,只经过2代,非B型即完全被B型替代。在棉花上,即使以非B型占87%、B型占13%开始共存竞争,经过225d后,非B型也完全被B型替代。这说明B型烟粉虱具有在短期内竞争取代浙江非B型烟粉虱的能力。经分析,B型除了寄主范围比非B型的宽这一点对其竞争有利外,较强的内在竞争潜能也是其能成功入侵并替代本地非B型烟粉虱的一个重要原因。     

基于地统计学的新疆棉田烟粉虱(Bemisia tabaci (Gennadius))危害动态与时空分布

应用地统计学(GS)的原理与方法研究了外来入侵有害生物烟粉虱(Bemisia tabaci (Gennadius))为害新疆棉田的时空动态,并与经典统计学进行对比分析.两种方法一致表明,烟粉虱成虫在空间上呈聚集分布的格局,而各时期成虫的聚集程度依据空间变异随机程度所占的比例不同而不同.经频次分布检验,以零频率法参数拟合的负二项分布来表达其空间分布型最为合适.运用GS的分析方法,进一步得到种群分布面积变化与扩散的趋势,并依此对昆虫种群的扩散模型进行模拟.棉田烟粉虱成虫在田间7～8月份均存在一定的空间相关性,随机程度为19.22%～49.99%;空间相关距离(相关程)在一个月内从32m急速增至6372m,随后在2000～3000m的范围波动.从整个发生过程看出,烟粉虱从越冬场所顺风侵入大田后,迁飞扩散在很大程度上受风向的影响,属于典型的借助风力扩散的昆虫,其在棉田的垂直分布则与吐鲁番地区独特的暖温带大陆性干旱荒漠气候特征有关.顺风扩散时多从棉株上部叶片开始危害, 而逆风扩散时从棉株中、下部叶片危害.烟粉虱在棉株上建立稳定种群后,中、下部虫口密度要略高于上部, 这是烟粉虱对吐鲁番地区特殊气候的适应.     

Fitness costs and morphological change of laboratory‐selected thiamethoxam resistance in the B‐type Bemisia tabaci (Hemiptera: Aleyrodidae)

Thiamethoxam has been used as a key insecticide to control the whitefly, B‐type Bemisia tabaci, for several years in China with no known cases of resistance in field populations. To evaluate the risk of resistance, a field population was collected and resistant strains were developed by exposure to thiamethoxam in the laboratory. After selection for 36 generation, a strain with 60‐fold resistance was successfully identified. Fitness analysis by constructing life tables, demonstrated that resistant B‐type whiteflies had obvious fitness disadvantages in their development and reproduction. The fitness of resistant B‐type whiteflies decreased dramatically, to only one‐half that of the susceptible strain. Some changes in the morphological characteristics of the resistant strain were observed. The lengths of first, second and third instars of the resistant strain were significantly smaller than those of the susceptible strain, and the width of the first and the fourth instars were also significantly smaller than in the susceptible strain. Our results suggest that the B‐type B. tabaci has the potential to develop high resistance to thiamethoxam, and that the resistance changed the morphology of the insects. The slow development of resistance and the lower fitness of resistant B. tabaci strains may result in a quick recovery of sensitivity when the population is no longer in contact with thiamethoxam in the field.     

Real‐time PCR quantification of Bemisia tabaci (Homoptera: Aleyrodidae) B‐biotype remains in predator guts

The cotton whitefly, Bemisia tabaci (Gennadius) B‐biotype, is fed on by a wide variety of generalist predators, but there is little information on these predator–prey interactions, especially under field conditions. In this study, a real‐time polymerase chain reaction (PCR) assay was developed to quantify B. tabaci B‐biotype remains in predator gut. The B. tabaci B‐biotype genomic DNA copy number was referred to the actual amount of BT1 isolate, the B. tabaci B‐biotype specific DNA fragment. The numbers of BT1 isolate in one B. tabaci B‐biotype egg, individual adult and a single red‐eyed nymph were 2.56 × 10³, 2.56 × 10⁴, and 1.29 × 10⁴ copies, respectively. When Propylaea japonica adults fed on one, two, four, eight or 16 red‐eyed nymphs, the detected numbers of BT1 isolate ranged from 2.77 × 10⁴ to 4.05 × 10⁵ copies, forming a strong linear relationship (R² = 0.9899). Following the consumption of two red‐eyed nymphs, prey DNA was detectable in 100% of P. japonica at t = 0, decreasing to 80.0% and 60.0% after 1–4 h and 8 h of digestion, respectively, with 3.36 × 10⁴–1.25 × 10³ BT1 isolate copies. The predation by field‐collected predators, 26 larvae of P. japonica, and of Harmonia axyridis each, Chrysopa spp. larvae (Chrysopa pallens and C. formosa, 18 individuals in total), and a single adult of Scymnus hoffmanni, 19 adults of Orius sauteri and nine adult spiders (Erigonnidium graminicolum and Neoscona doenitzi), on B. tabaci B‐biotype were quantified. Of the 99 analysed predator individuals, 3.65 × 10²–4.60 × 10⁵ copies of BT1 isolate, equivalent to 0.8–18.8 red‐eyed nymphs were detected. These results suggest that TaqMan real‐time PCR technology may provide a rapid and sensitive method for quantifying B. tabaci B‐biotype remains in predator guts and will be invaluable in assessing the food web relationship between prey and arthropod predators.     

Discrimination of cassava-associated Bemisia tabaci in Africa from polyphagous populations, by PCR–RFLP of the internal transcribed spacer regions of ribosomal DNA

Restriction fragment length polymorphism (RFLP) analysis of the ribosomal DNA internal transcribed spacer regions of Bemisia tabaci was used to distinguish cassava‐associated populations from other host‐associated populations. Endonuclease restriction profile analysis indicated that cassava‐associated populations from Africa represent a distinct group, with a significant level of separation into subgroups that were not linked to geographical origin. Analysis of molecular variance (amova ) revealed that a high proportion of the total genetic variation (47%) was attributable to among‐population differences within the host‐associated groups. Principal coordinate analysis supported the differentiation between the cassava and the non‐cassava group, a result which was in agreement with the cluster analysis of the restriction fragment profile. Internal transcribed spacer RFLP markers, especially SmaI, identified in this study can be used to monitor the spread of B. tabaci biotypes, especially of the more virulent biotype B that has so far not been reported in the cassava‐growing belt of Africa.