烟粉虱及温室白粉虱为害后对后续粉虱种群的影响

【目的】分析烟粉虱Bemisia tabaci(Gennadius)或温室白粉虱Trialeurodes vaporariorum(Westwood)为害番茄后对后续温室白粉虱和烟粉虱生长、发育、成虫寿命和繁殖等产生的促进或抑制作用,可为明确寄主植物番茄介导的温室白粉虱-烟粉虱的种间互作,开展粉虱的科学防控提供科学依据。【方法】通过在寄主植物番茄叶片上先将B型烟粉虱或温室白粉虱按不同的顺序、时间间隔分开接种,再系统观察后续接上的两种粉虱生长、发育、繁殖等种群参数的变化。【结果】(1)先期接上烟粉虱对后续温室白粉虱的发育、寿命、产卵量有明显的促进作用;这种作用需烟粉虱的持续诱导,若去掉烟粉虱,其对温室白粉虱的促进作用即消失;先期接上温室白粉虱可缩短后续烟粉虱伪蛹期,但温室白粉虱的持续存在不利于烟粉虱的产卵,且明显降低烟粉虱的内禀增长率和净增殖率。(2)先后在番茄上同时接上温室白粉虱可降低后续烟粉虱的单雌产卵量和雌、雄虫的成虫寿命;先后同时接种烟粉虱却显著地增加了温室白粉虱的单雌产卵量;烟粉虱的提前存在降低了后续烟粉虱单雌产卵量。(3)但两种粉虱之间作用存在着一定的时间滞后性。烟粉虱对温室白粉虱的发育、寿命、产卵量产生的促进作用大约在其卵期后的一段时间才能显现出来;温室白粉虱对烟粉虱伪蛹期的缩短作用也需提前诱导;而温室白粉虱对温室白粉虱的促进作用相当滞后,直到成虫期才显现出来。【结论】B型烟粉虱和温室白粉虱之间可通过寄主番茄产生相互影响,前期烟粉虱为害可显著促进后续温室白粉虱卵和若虫的发育,而前期温室白粉虱为害对烟粉虱的发育不利。     

B型烟粉虱与温室白粉虱不同虫态的碱性磷酸酶性质比较

为了探明B型烟粉虱Bemisia tabaci B-biotype 和温室白粉虱Trialeurodes vaporariorum体内的碱性磷酸酶(alkaline phosphatase,ALP)在两者竞争替代中所起的作用,以对硝基苯磷酸二钠 (pNPP)为底物,采用个体测定和群体测定的方法,研究比较了2种粉虱不同虫态中该酶的性质。结果表明：2种粉虱的碱性磷酸酶比活力在整个发育历期均逐渐增加,成虫期达到最大。温室白粉虱2至4龄若虫（伪蛹）期的碱性磷酸酶比活力分别是B型烟粉虱对应龄期酶比活力的2.58、2.68和3.14倍; B型烟粉虱雌雄成虫的碱性磷酸酶比活力分别是温室白粉虱雌雄成虫酶比活力的1.24和1.26倍,且2种粉虱雌虫的酶比活力显著大于其雄虫。2种粉虱2龄若虫到成虫的碱性磷酸酶最适pH均为7.8,最适温度均为47℃;在1龄若虫中均未能检测到该酶活性。测定并比较2种粉虱不同虫态碱性磷酸酶动力学特征参数的结果显示,温室白粉虱碱性磷酸酶在3、4龄若虫的亲和力以及在2, 3, 4龄若虫的酶蛋白浓度均显著大于B型烟粉虱的对应值,而在成虫期2种粉虱的亲和力、酶蛋白浓度无差异,B型烟粉虱的活化能显著小于温室白粉虱。据此推测,B型烟粉虱利用碱性磷酸酶在若虫期进行组织骨化和生长发育不如温室白粉虱,但羽化为成虫后利用其进行解毒代谢则可能强于温室白粉虱。     

一种改进的粉虱成虫生物测定方法

生物测定是检测害虫抗药性的一项重要技术。利用100mL插口圆底聚丙烯离心管对现在应用较多的粉虱成虫生物测定方法——琼脂保湿浸叶法进行了改进。改进后的方法不影响粉虱成虫的持续取食,具有操作简单、结果重复性好及无需对成虫麻醉等优点。同时发现茄子叶片对于B型烟粉虱Bemisia tabaci(Gennadius)B-biotype和温室白粉虱Trialeurodes vaporariorum(Westwood)成虫均是一种非常适合的生测材料。利用该方法分3次独立测定了烯啶虫胺对B型烟粉虱和温室白粉虱混合日龄成虫的毒力,结果具有很好的重复性。     

B型烟粉虱和温室白粉虱在温度逆境下的生存特性比较

为了明确B型烟粉虱和温室白粉虱在温度逆境下的生存特性对其种群发展的影响,通过进行高温和低温暴露试验,研究了B型烟粉虱和温室白粉虱卵、伪蛹、成虫在37℃,39℃,41℃,43℃,45℃下暴露1～2 h后的存活率,以及这两种粉虱卵、2～3龄若虫、伪蛹和成虫在2℃下暴露1～12 d后的存活率。结果表明：两种粉虱的卵、伪蛹和成虫在37℃～45℃下暴露1～2 h,其存活率均随着温度的上升而降低;但在相同处理条件下B型烟粉虱3种供试虫态的存活率要高于温室白粉虱。B型烟粉虱在2℃下暴露2～12 d,各供试虫态的存活率迅速下降,卵、2～3龄若虫、伪蛹在2℃下暴露12 d后均不能存活,成虫在2℃下暴露4 d后也全部死亡;而温室白粉虱卵、伪蛹在2℃下暴露12 d后其存活率还能超过45%,成虫在2℃下暴露7 d后仍有80.9%能够存活。结果说明,B型烟粉虱和温室白粉虱对温度逆境的适应性存在差异,B型烟粉虱对高温的适应性要高于温室白粉虱;温室白粉虱对高温敏感,但对低温的适应性要显著高于B型烟粉虱。据此推测,两种粉虱对温度逆境适应性的差异是导致其种群发生存在差异的一个重要原因。     

烟草品种和温度对B型烟粉虱和温室白粉虱生物学参数的影响

在室内20℃、26℃和30℃三个设定温度条件下,研究了CF-965、NC-89和NC-82三个烟草品种对B型烟粉虱和温室白粉虱生长发育和繁殖的影响.结果表明:26℃时两种粉虱在CF-965、NC-89和NC-82三个烟草品种上的发育历期没有明显差异,但温室白粉虱卵期和若虫期的发育历期较烟粉虱依次分别延长了1.32d、2.81d和 2.04d.在CF-965上,B型烟粉虱卵期和若虫期的总存活率和平均单雌产卵量分别为54.19%和55.17粒,温室白粉虱仅分别为31.99%和39.17粒;在NC-89上,B型烟粉虱卵期和若虫期的总存活率和平均单雌产卵量分别为23.47%和37.50粒,温室白粉虱仅分别为15.75%和17.25粒;两种粉虱在NC-82上卵期和若虫期的总存活率和平均单雌产卵量差异不大,表明B型烟粉虱对CF-965和NC-89的适应能力较温室白粉虱强.两种粉虱在20℃时的发育历期最长,32℃时最短,26℃时居中.20°C时烟粉虱卵期和若虫期的存活率20.96%为最低,26℃和32℃时分别达54.19% 和53.29%;而温室白粉虱在32℃条件下存活率11.66%为最低,20℃和 26℃时分别达31 34% 和31.99%.温室白粉虱在20℃时产卵量大于烟粉虱, 在26℃时的产卵量小于烟粉虱,30℃时则不能产卵,但烟粉虱成虫30℃时的产卵量仍可达21.06粒.由此说明烟粉虱在较高温度条件下的适应能力显著强于温室白粉虱.山东烟区烟草生长中期的田间平均温度多在22～30℃,适合B型烟粉虱的发生,应引起高度重视.     

辽宁省冬季温室内越冬粉虱伪蛹的种类鉴定及烟粉虱体内番茄黄化曲叶病毒的检测

为了揭示辽宁省冬季温室内越冬粉虱伪蛹的种类及烟粉虱Bemisia tabaci （Gennadius）携带番茄黄化曲叶病毒（tomato yellow leaf curl virus, TYLCV）情况, 于2012年1月份在辽宁省不同县市区的温室作物上采集了17份粉虱伪蛹样品（每样品含30头粉虱伪蛹） , 镜检鉴别粉虱种类并利用mtCOI基因对烟粉虱生物型进行了鉴定; 检测了烟粉虱携带TYLCV情况并对其PCR扩增产物进行了测序分析。结 果表明： 辽宁省冬季温室内存在越冬温室白粉虱Trialeurodes vaporariorum （Westwood）与烟粉虱。17份粉虱样品中, 11份样品为烟 粉虱样品, 6份样品为温室白粉虱和烟粉虱混合样品。混合样品中, 温室白粉虱仅在锦州凌海（LH）样品中占优势。17份烟粉虱样品（包 括混合样品）中, 仅有4份样品为B型与Q型混合样品, 其他13份样品烟粉虱生物型均为Q型。17份烟粉虱样品中有3份Q型烟粉虱样品检测 到TYLCV, 系统树分析进一步证实该病毒是TYLCV。调查结果为辽宁省粉虱与TYLCV的早期测报和防控提供了科学依据。     

番茄黄化曲叶病毒对温室白粉虱适合度的影响

【目的】温室白粉虱Trialeurodes vaporariorum(Westwood)是为害北方地区花卉蔬菜的主要粉虱种类,烟粉虱Bemisia tabaci(Gennadius)则在近些年逐渐频繁的花卉贸易活动中扩散至黑龙江省部分地区并取代了白粉虱成为当地温室害虫的优势物种,番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)是烟粉虱传播的一种重要双生病毒,对作物的危害十分严重,然而该病毒对本地白粉虱的影响及对烟粉虱与白粉虱种间竞争关系的影响尚待研究。【方法】本研究观察记录感染番茄黄化曲叶病毒的番茄植株上温室白粉虱和烟粉虱的种群动态及番茄形态和部分生理指标的变化。【结果】结果表明:1)有烟粉虱滋生的番茄植株矮小,根系发达;2)有白粉虱滋生的番茄略微矮粗,影响较小;3)感染病毒的番茄矮粗或矮小,根部无明显变化;4)染毒带虫相对于带虫处理而言,在白粉虱试验中,番茄植株矮小,根系生物量也骤减;与此相反,在烟粉虱试验中,番茄的反应相对缓和;5)此外,不同酶类对植株染毒的响应不同:Ach E酶活高不利于植株,而GST酶活高则有利于植株。【结论】总体而言:烟粉虱单独作用很不利于苗期番茄,白粉虱对苗期番茄没有明显的直接影响;而感染病毒会缓解烟粉虱对番茄的强烈刺激,而加重白粉虱对番茄的作用,即染毒使带白粉虱的苗期番茄生长发育受到明显抑制。与番茄变化情况相对应的是,染毒番茄上烟粉虱产卵较少,但在发育前期(从卵到伪蛹)存活率较高;染毒番茄上白粉虱产卵较多,但在前期存活率较低。本研究可为高纬度地区粉虱综合防控提供参考。     

三种粉虱触角、口针及跗节感受器的超微结构观察

对烟粉虱Bemisia tabaci、温室白粉虱Trialeurodes vaporariorum和禾粉虱Aleurocybotus indicus3种粉虱的触角、口针及跗节感受器的超微结构进行了观察。利用扫描电镜观察了烟粉虱、温室白粉虱和禾粉虱3种粉虱触角、口针及跗节化学感受器的类型和分布,进行了比较。3种粉虱触角上都有刚毛感受器、锥形感受器2种感受器和微毛,烟粉虱触角还有钟形感受器和腔锥感受器;温室白粉虱和禾粉虱触角均没有发现钟形感受器;3种粉虱的口针和跗节上均只有刚毛感受器和微毛,且形态结构相似。     

不同邻作作物对番茄田间烟粉虱种群动态的影响

为明确不同邻作模式下番茄田间烟粉虱的种群动态与空间分布格局,于云南省昆明市选取与西葫芦、黄瓜、玫瑰等作物邻作的番茄田及单作番茄田为研究样地,通过五点取样法进行调查,采用四分位法对不同作物邻作的番茄田间烟粉虱种群动态进行分析。结果表明:在烟粉虱的主要发生期,番茄单作田间烟粉虱种群数量最高,雌成虫达26.69头·叶^-1,雄成虫达6.74头·叶^-1,若虫达16.47头·叶^-1,与西葫芦、黄瓜、玫瑰邻作的番茄田间烟粉虱种群数量均较低,且与黄瓜邻作的番茄田间烟粉虱主要发生时期较短,发生高峰期最晚。在整个调查期内,所有邻作模式的番茄田间烟粉虱雌虫数量均高于雄虫;在烟粉虱的主要发生期,与西葫芦邻作的番茄田间烟粉虱性比最高,达8.20。不同邻作模式番茄田间烟粉虱成虫数量均在番茄植株上部叶片最多,下部叶片最少;烟粉虱若虫数量均在番茄植株中部叶片最多,上部叶片最少。在番茄植株不同部位,烟粉虱成虫和若虫数量均在番茄单作田间最大,在邻作玫瑰的番茄田最少。可考虑在烟粉虱严重为害的番茄种植区附近种植玫瑰等花卉,以降低烟粉虱的种群数量。     

甘蓝与黄瓜寄主上B型烟粉虱和温室白粉虱蜜露糖分、氨基酸和挥发物组分的比较分析

粉虱蜜露是粉虱寄生性天敌搜索寄主的主要利它素源。应用离子色谱分别对甘蓝与黄瓜上B型烟粉虱(Bemisia tabaci B-biotype)蜜露以及黄瓜上温室白粉虱Trialeurodes vaporariorum蜜露的接触性利它素糖和氨基酸组分和含量进行了比较研究。结果表明：2种粉虱在不同寄主植物上的蜜露均富含糖和氨基酸,其中糖含量占绝对优势,甘蓝上B型烟粉虱蜜露、黄瓜上B型烟粉虱蜜露和黄瓜上温室白粉虱蜜露中的糖含量分别是相应氨基酸含量的42.5、2.6和5.4倍,其中糖类物质中又以寡糖含量占绝对优势,分别占89.3%、81.7%和88.2%。不同寄主植物和粉虱种类显著影响蜜露中糖和氨基酸的组成和含量。其中,甘蓝上B型烟粉虱蜜露中的寡糖以二糖占优势,占97.3%;二糖中又以蔗糖异构糖和松二糖占优势,分别占52.7%和35.4%。黄瓜上B型烟粉虱蜜露和温室白粉虱蜜露寡糖中以三糖和四糖占优势,分别占73.1%和85.4%;优势糖水苏（四）糖和松三糖分别占40.3%和 26.2%及49.9%和27.0%。甘蓝上B型烟粉虱蜜露中氨基酸以丙氨酸占优势,含量为66.5%;而黄瓜上B型烟粉虱及温室白粉虱蜜露中氨基酸以甘氨酸含量最高,分别占氨基酸总量的38.2%和51.7%。应用GC-MS对甘蓝上B型烟粉虱蜜露和黄瓜上温室白粉虱蜜露挥发物组分的鉴定结果显示,两种粉虱蜜露中共同含有的主要挥发物为邻苯二甲酸二（2-乙基）己酯。     

Interspecific interactions between Bemisia tabaci biotype B and Trialeurodes vaporariorum (Hemiptera: Aleyrodidae)

Bemisia tabaci (Gennadius) biotype B and Trialeurodes vaporariorum (Westwood) are invasive whitefly species that often co-occur on greenhouse-grown vegetables in northern China. Although B. tabaci biotype B has been present in China for a relatively short period of time, it has become dominant over T. vaporariorum. We studied the interspecific competitive interactions between the two species in single or mixed cultures at 24 ± 1 °C, 40 ± 5% RH, and L14:D10 h photoperiod. Female longevity on tomato was not significantly different between species, but B. tabaci reproduced 4.3 to 4.9 fold more progeny. The ratio of female to male progeny in both instances was greater for B. tabaci. When cultured on tomato, cotton, and tobacco, B. tabaci developed 0.8, 3.3, and 4.7 d earlier in single culture, and 1.8, 3.9, and 4.3 d earlier in mixed culture. B. tabaci displaced T. vaporariorum in four, five and six generations when the initial ratios of B. tabaci to T. vaporariorum were 15:15, 20:10, or 10:20 on tomato. Populations of B. tabaci were 2.3 fold higher than that of T. vaporariorum on tomato plants for seven consecutive generations in single culture. B. tabaci performed better in development, survival, fecundity, and female ratio. We conclude that B. tabaci could displace T. vaporariorum in as short as four generations in a controlled greenhouse environment when they start at equal proportions. Warmer greenhouse conditions and an increase in total greenhouse area could be contributing factors in the recent dominance of B. tabaci.     

Searching behaviour of Encarsia formosa as mediated by colour and honeydew

The habitat- and host-searching behaviour of female Encarsia formosa Gahan (Hymenoptera: Aphelinidae) was assessed using an airflow olfactometer and a filter paper test. Responses to different odour cues, colours, host-produced honeydew, non-host honeydew and single carbohydrates were determined. The parasitoid was not attracted to or arrested by odours emanating from clean tobacco leaves, tobacco leaves heavily infested with the host Trialeurodes vaporariorum (Westwood) (Homoptera: Aleyrodidae) and covered with honeydew, or honeydew alone. However, E. formosa females showed a significant response to green light transmitted through a tobacco leaf. The yellow part of the spectrum was partly responsible for this response. Thus, the long-range orientation is random with respect to the presence of hosts. Filter paper tests showed that the short-range searching behaviour is influenced by water soluble, non-volatile contact-kairomones contained in the host-produced honeydew. Contact with honeydew excreted by L3/L4 T. vaporariorum resulted in longer searching times than honeydew from adult T. vaporariorum or L3/L4 Bemisia tabaci Gennadius (Homoptera: Aleyrodidae). No difference was found between the response to honeydew excreted by adult and L3/L4 B. tabaci. The parasitoids' response to honeydew was unaffected by the host plant on which the whiteflies had fed. Non-host honeydew and single carbohydrates also affected the searching behaviour of E. formosa but to a lower extent than host honeydew. The possible differences in the carbohydrate and amino acid composition of the honeydew excreted by different life-stages of T. vaporariorum and B. tabaci are discussed.     

Feeding preference of Macrolophus caliginosus (Heteroptera: Miridae) on Bemisia tabaci and Trialeurodes vaporariorum (Homoptera: Aleyrodidae)

A study of predation choices of Macrolophus caliginosus Wagner (Heteroptera: Miridae) late instars and adults, when offered various developmental stages (eggs and nymphs) of the recently established whitefly Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae), was made based on two preference indices. In addition, prey choices of late instars when presented with three ratios of Trialeurodes vaporariorum (Westwood) (Homoptera: Aleyrodidae) and B. tabaci at a similar developmental stage (eggs, young or late instars) were assessed. M. caliginosus preferred older nymphs of B. tabaci than any other stage. It also chose T. vaporariorum over B. tabaci, unless the latter consisted of > 75% of the available prey. These results suggested that M. caliginosus might interfere with parasitoids such as Encarsia, Eretmocerus, or Amitus spp. because all three species emerge from the host pupal case. Furthermore, in mixed infestations, M. caliginosus preference for T. vaporariorum might either negatively affect the control of B. tabaci, or, contrarily, enhance the predator population, before a B. tabaci outbreak occurs in the greenhouse.     

两种入侵粉虱热激基因Hsp70的克隆及温度胁迫下的种间差异表达

为了检测B型烟粉虱和温室白粉虱在受到温度胁迫时热激基因Hsp70的差异表达谱,克隆了2种粉虱Hsp70基因的全长cDNA序列,并用实时定量PCR的方法分析了在不同温度条件下该基因的表达谱。结果表明:B型烟粉虱和温室白粉虱的Hsp70基因BTH-sp70和TVHsp70(GenBank登录号分别为DQ093385和EU861391),在5’-非编码区(UTR)中有类似TATA-box样元件;在3’-UTR中有poly(A)信号AATAA和富含A-T区;根据基因开放阅读框序列(ORF)推导的氨基酸序列均含有全部3个Hsp70蛋白家族高度保守的基序。在整个检测的温度范围内(-19℃～46℃),诱导温室白粉虱Hsp70表达的起始温度(Ton)或最大温度(Tmax)要比B型烟粉虱低2.5℃～7.5℃。这些结果表明,所克隆的基因属于有功能的温度诱导型Hsp70基因;在基因表达水平上,温室白粉虱比B型烟粉虱更耐冷,而后者更耐热;Hsp70的Ton(或者Tmax)能代表这2种粉虱的温度耐受能力。本研究结果在一定程度上解释了自然界中这2种粉虱种群地理分布和季节发生差异的原因。     

Host-parasite interactions between whiteflies and their parasitoids

There is relatively little information available concerning the physiological and biochemical interactions between whiteflies and their parasitoids. In this report, we describe interactions between aphelinid parasitoids and their aleyrodid hosts that we have observed in four host-parasite systems: Bemisia tabaci/Encarsia formosa, Trialeurodes vaporariorum/E. formosa, B. tabaci/Eretmocerus mundus, and T. lauri/Encarsia scapeata. In the absence of reported polydnavirus and teratocytes, these parasitoids probably inject and/or produce compounds that interfere with the host immune response and also manipulate host development to suit their own needs. In addition, parasitoids must coordinate their own development with that of their host. Although eggs are deposited under all four instars of B. tabaci, Eretmocerus larvae only penetrate 4th instar B. tabaci nymphs. A pre-penetrating E. mundus first instar was capable of inducing permanent developmental arrest in its host, and upon penetration stimulated its host to produce a capsule (epidermal in origin) in which the parasitoid larva developed. T. vaporariorum and B. tabaci parasitized by E. formosa initiated adult development, and, on occasion, produced abnormal adult wings and eyes. In these systems, the site of parasitoid oviposition depended on the host species, occurring within or pressing into the ventral ganglion in T. vaporariorum and at various locations in B. tabaci. E. formosa's final larval molt is cued by the initiation of adult development in its host. In the T. lauri-E. scapeata system, both the host whitefly and the female parasitoid diapause during most of the year, i.e., from June until the middle of February (T. lauri) or from May until the end of December (E. scapeata). It appears that the growth and development of the insects are directed by the appearance of new, young foliage on Arbutus andrachne, the host tree. When adult female parasitoids emerged in the spring, they laid unfertilized male-producing eggs in whiteflies containing a female parasitoid [autoparasitism (development of male larvae utilizing female parasitoid immatures for nutrition)]. Upon hatching, these male larvae did not diapause, but initiated development, and the adult males that emerged several weeks later mated with available females to produce the next generation of parasitoid females. Thus, the interactions that exist between whiteflies and their parasitoids are complex and can be quite diverse in the various host-parasitoid systems.     

温室粉虱和烟粉虱3个隐种中热激蛋白基因hsp70和hsp90含量的比较分析

【目的】昆虫适应新环境的能力与其对温度的耐受能力密切相关。热激蛋白HSP70和HSP90具有提高生物体温度耐受性的功能。烟粉虱Bemisia tabaci (Gennadius)不同隐种和不同种粉虱对温度的适应能力有差异,这与它们的热激蛋白基因拷贝数的差异可能相关。【方法】利用实时荧光定量PCR方法,检测入侵型烟粉虱MED隐种和MEAM1隐种、本地型烟粉虱AsiaⅡ1隐种以及温室粉虱Trialeurodes vaporariorum (Westwood)基因组DNA中热激蛋白基因hsp70和hsp90的拷贝数。【结果】不同种类的粉虱和烟粉虱不同隐种体内的hsp70和hsp90的含量均有较大差异,其中hsp70和hsp90拷贝数在入侵型烟粉虱MED和MEAM1隐种中含量较其他两种均高,而在土著种AsiaⅡ1隐种中含量最低,在温室粉虱中居中。此外,相同物种雌雄成虫hsp70和hsp90的拷贝数也不同,雌虫hsp70和hsp90拷贝数约为雄虫的2倍。【结论】不同种粉虱及烟粉虱不同隐种的hsp70和hsp90的拷贝数可能与其耐热性差异相关。本研究为解释不同种粉虱、烟粉虱不同隐种及其不同性别的耐热性差异机制提供了进一步的依据。     

Within-plant distribution and sampling of single and mixed infestations of Bemisia tabaci and Trialeurodes vaporariorum (Homoptera: Aleyrodidae) in winter tomato crops

In several areas of Spain, the greenhouse whitefly, Trialeurodes vaporariorum (Westwood), and the sweet potato whitefly, Bemisia tabaci (Gennadius), coexist in tomato, Lycopersicon esculentum Miller. For integrated pest management decision-making, it is important to know the abundance of each species, because they exhibit different abilities to transmit viruses, are susceptible to different biological control agents, and have different responses to insecticides. This study was conducted to provide information on the vertical distribution of T. vaporariorumn and B. tabaci in tomato plants grown in greenhouses in winter and to determine the optimal sampling unit and the sample size for estimating egg and nymphal densities of both whitefly species. Eggs of T. vaporariorum were mainly located on the top stratum of the plant, whereas B. tabaci eggs were mainly found on the middle stratum. Nymphs of both species mainly concentrated in the bottom stratum of the plant. When pest abundance and low relative variation were considered, the bottom stratum was selected as the most convenient for sampling nymphs of both whitefly species. Conversely, the same two criteria indicated that either the top or the middle strata could be used when sampling T. vaporariorum and B. tabaci eggs. Several different sampling units were compared to optimize the estimation of nymphal and egg densities in terms of cost efficiency. One disk (1.15 cm in diameter) per leaflet collected from the top stratum of the tomato plant was the most efficient sampling unit for simultaneously estimating the egg densities of the two whitefly species.