RNA interference-mediated tolerance to whitefly (Bemisia tabaci) in genetically engineered tomato

Plant Cell, Tissue and Organ Culture (PCTOC) - Whitefly (Bemisia tabaci) is a polyphagous insect that causes huge damage in several horticultural crops, including tomato, by sucking nutrients from...     

Differential Responses of the Whitefly Bemisia tabaci Symbionts to Unfavorable Low and High Temperatures

The whitefly Bemisia tabaci complex contains many cryptic species, of which the Middle East-Asia Minor 1 (MEAM1) and Mediterranean (MED) are notorious invasive pests. In our field-collected whitefly samples, MEAM1 harbors an obligate primary symbiont “Candidatus Portiera aleyrodidarum” and two secondary symbionts, “Candidatus Hamiltonella defensa” and Rickettsia sp., whereas MED has only “Ca. Portiera aleyrodidarum” and “Ca. Hamiltonella defensa.” Both “Ca. Portiera aleyrodidarum” and “Ca. Hamiltonella defensa” are intracellular endosymbionts residing in the bacteriomes, whereas Rickettsia sp. has a scattered distribution throughout the host body cavity. We examined responses of these symbionts to adverse temperatures as well as survival of the host insects. After cold treatment at 5 or 10 °C or heat treatment at 35 or 40 °C for 24 h, respectively, the infection rates of all symbionts were not significantly decreased based on diagnosis PCR. However, quantitative PCR assays indicated significant reduction of “Ca. Hamiltonella defensa” at 40 °C, and the reduction became greater as the duration increased. Compared with “Ca. Hamiltonella defensa,” “Ca. Portiera aleyrodidarum” was initially less affected in the first day but then showed more rapid reduction at days 3–5. The density of Rickettsia sp. fluctuated but was not reduced significantly at 40 °C. Meanwhile, the mortality rates of the host whiteflies elevated rapidly as the duration of exposure to heat treatment increased. The differential responses of various symbionts to adverse temperatures imply complex interactions among the symbionts inside the same host insect and highlight the importance of taking the whole bacterial community into account in studies of symbioses.     

Whitefly Bemisia tabaci (Homoptera: Aleyrodidae) infestation on cassava genotypes grown at different ecozones in Nigeria

Large-scale screening of cassava, Manihot esculenta Crantz, genotypes for resistance to infestation by whitefly Bemisia tabaci Gennadius, the vector of cassava mosaic geminiviruses, is limited. A range of new cassava elite clones were therefore assessed for the whitefly infestation in the 1999/2000 and 2000/2001 cropping seasons in experimental fields of International Institute of Tropical Agriculture, Ibadan, Nigeria. On each scoring day, between 0600 and 0800 hours when the whiteflies were relatively immobile, adult whitefly populations on the five topmost expanded leaves of cassava cultivars were counted. All through the 6-mo scoring period, there was a highly significant difference in whitefly infestation among the new cassava elite clones. Vector population buildup was observed in Ibadan (forest-savanna transition zone) and Onne (humid forest), 2 mo after planting (MAP). Mean infestation across cassava genotypes was significantly highest (16.6 whiteflies per plant) in Ibadan and lowest in Zaria (0.2). Generally, whitefly infestation was very low in all locations at 5 and 6 MAP. During this period, cassava genotypes 96/1439 and 91/02324 significantly supported higher infestations than other genotypes. Plants of 96/1089A and TMS 30572 supported the lowest whitefly infestation across cassava genotypes in all locations. The preferential whitefly visitation, the differences between locations in relation to whitefly population, cassava mosaic disease, and the fresh root yield of cassava genotypes are discussed.     

寄生烟粉虱的桨角蚜小蜂新记录种

记述寄生烟粉虱Bemisia tabaci(Gennadius)的桨角蚜小蜂属Eretmocerus Haldeman 4个中国新记录种:酋长桨角蚜小蜂E.emiratus,占桥桨角蚜小蜂E.furuhashii,海氏桨角蚜小蜂E.hayati和刻盾桨角蚜小蜂E$CUlpturatus,以及1个大陆新记录种:黑盾桨角蚜小蜂E.melanoscutus.另外,还提供了中国桨角蚜小蜂属分种检索表.     

The Efficacy of Yellow Tape Formulation of Pyriproxyfen against the Sweet Potato Whitefly,Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae)

A tape formulation of pyriproxyfen was prepared by coating the chemical onto yellow plastic film. We evaluated the transovarial ovicidal activity of the tape formulation against Bemisia tabaci and investigated the optimal application method and efficacy in a greenhouse. The tape formulation showed high ovicidal activity of over 80% mortality against B. tabaci for 4 days by a 30-second compulsory contact test. The formulation controlled the B. tabaci population on tomatoes by both “sideways spreading application” and “vertical hanging application”. “Sideways spreading application” was considered to be more practical, although there was no significant difference in efficacy between the two application methods. In a large-scale greenhouse trial of “sideways spreading application”, the tape formulation suppressed the B. tabaci population for 77 days. The results suggested that the tape formulation is promising as a tool for IPM because of its high efficacy and lower environmental impact.     

Tissue-specific gene silencing by RNA interference in the whitefly Bemisia tabaci (Gennadius)

The hemipteran whitefly Bemisia tabaci (Gennadius) species complex and the plant viruses they transmit pose major constraints to vegetable and fiber production, worldwide. The whitefly tissue- and developmental-specific gene expression has not been exhaustively studied despite its economic importance. In 2002, a functional genomic project was initiated, which generated several thousands expressed sequence tags (ESTs) and their sequence. This project provides the basic information to design experiments aimed at understanding and manipulating whitefly gene expression. In this communication, for the first time we provide evidence that the RNA interference mechanism discovered in many organisms, including in Hemiptera, is active in B. tabaci. By injecting into the body cavity long double-stranded RNA (dsRNA) molecules, specifically directed against genes uniquely expressed in the midgut and salivary glands, we were able to significantly inhibit the expression of the targeted mRNA in the different organs. Gene expression levels in RNAi-silenced whiteflies were reduced up to 70% compared to whiteflies injected with buffer or with a green fluorescent protein (GFP)-specific dsRNA. Phenotypic effects were observed in B. tabaci ovaries following dsRNA targeting the whitefly Drosophila chickadee homologue. Disruption of whitefly gene expression opens the door to new strategies aimed at curbing down the deleterious effects of this insect pest to agriculture.     

烟粉虱对寄主植物的适应性

就寄主植物营养对烟粉虱的影响,以及烟粉虱在遭受寄主植物营养胁迫和植物次生代谢物质刺激时的生理适应机制进行了综述.寄主植物体内的氮营养物质对烟粉虱有显著的影响,烟粉虱也可以通过多种方式来适应寄主植物体内氮营养物质的变化.当植物体内氮营养不足时,烟粉虱通过利用体内的共生菌、改变代谢方式、诱导寄主植物改变同化产物形成方向等方式来确保生长发育所必须的氮营养物质.烟粉虱还通过体内乙酰胆碱酯酶和羧酸酯酶来适应植物次生代谢物质的改变.另外,我们还对烟粉虱具有较强暴发潜能的生理机制进行了分析.     

DNA Barcoding of Bemisia tabaci Complex (Hemiptera: Aleyrodidae) Reveals Southerly Expansion of the Dominant Whitefly Species on Cotton in Pakistan

Background

Although whiteflies (Bemisia tabaci complex) are an important pest of cotton in Pakistan, its taxonomic diversity is poorly understood. As DNA barcoding is an effective tool for resolving species complexes and analyzing species distributions, we used this approach to analyze genetic diversity in the B. tabaci complex and map the distribution of B. tabaci lineages in cotton growing areas of Pakistan.

Methods/Principal Findings

Sequence diversity in the DNA barcode region (mtCOI-5′) was examined in 593 whiteflies from Pakistan to determine the number of whitefly species and their distributions in the cotton-growing areas of Punjab and Sindh provinces. These new records were integrated with another 173 barcode sequences for B. tabaci, most from India, to better understand regional whitefly diversity. The Barcode Index Number (BIN) System assigned the 766 sequences to 15 BINs, including nine from Pakistan. Representative specimens of each Pakistan BIN were analyzed for mtCOI-3′ to allow their assignment to one of the putative species in the B. tabaci complex recognized on the basis of sequence variation in this gene region. This analysis revealed the presence of Asia II 1, Middle East-Asia Minor 1, Asia 1, Asia II 5, Asia II 7, and a new lineage “Pakistan”. The first two taxa were found in both Punjab and Sindh, but Asia 1 was only detected in Sindh, while Asia II 5, Asia II 7 and “Pakistan” were only present in Punjab. The haplotype networks showed that most haplotypes of Asia II 1, a species implicated in transmission of the cotton leaf curl virus, occurred in both India and Pakistan.

Conclusions

DNA barcodes successfully discriminated cryptic species in B. tabaci complex. The dominant haplotypes in the B. tabaci complex were shared by India and Pakistan. Asia II 1 was previously restricted to Punjab, but is now the dominant lineage in southern Sindh; its southward spread may have serious implications for cotton plantations in this region.     

Bemisia tabaci (Homoptera: Aleyrodidae) biotypes in India

Host plant performance, esterase, and virus transmission tests revealed cassava-strain and sweetpotato-strain populations of whitefly Bemisia tabaci (Gennadius) biotypes in India. Individuals from the sweetpotato-reared population did not breed on cassava, Manihot esculenta Crantz, and the cassava-strain-reared individuals failed to develop on sweetpotato, Ipomoea batatus (L.) Lam. Eggplant, Solanum melongena L., and tobacco, Nicotiana tabacum L., were common hosts for both biotypes. The cassava-strain whiteflies but not the sweetpotato-reared whiteflies successfully transmitted cassava mosaic virus from disease-infected cassava seedlings to healthy cassava seedlings. Presence of biotypes in B. tabaci is reported for the first time from India.     

烟粉虱对四种蔬菜寄主的选择性

为探讨利用趋避作用防治烟粉虱的可能性,进行了3方面试验：1）烟粉虱Bemisia tabaci (Gennadius) 对黄瓜、花椰菜、油菜和莴苣等4种蔬菜的寄主选择性; 2）互喷烟粉虱不同嗜好的寄主植物汁液对其寄主选择性的影响; 3）不同间作方式对黄瓜叶上烟粉虱成虫密度的影响。结果表明,烟粉虱对供试植物有明显的选择性,其嗜好程度为花椰菜>黄瓜>油菜>莴苣。喷施莴苣和花椰菜汁液对烟粉虱的寄主选择性均有显著影响,喷施莴苣原液后,花椰菜和黄瓜上的成虫数分别减少79.6%和87.4%,花椰菜上的着卵量减少84.3%;喷施花椰菜原液后,莴苣上的成虫数和着卵量分别增加82.4%和79.2%,表明不同蔬菜中的内含物是引起烟粉虱对寄主选择性的重要原因。在黄瓜中间作莴苣可使黄瓜叶上烟粉虱成虫数平均减少76.5%;间作花椰菜时,通过诱集作用,也可使黄瓜叶上的成虫数平均降低69.7%。这些结果为利用喷施蔬菜汁液和间作方式控制烟粉虱危害提供了理论依据。     

Modelling temperature-dependent bionomics of Bemisia tabaci (Q-biotype)

Abstract.  The influence of temperature (17, 21, 25, 30 and 35 °C) on life-history traits of a Q-biotype Bemisia tabaci population on tomato is studied. Temperature-dependent relationships are characterized for immature developmental rate, immature survival, fecundity, longevity and intrinsic rate of increase. Development time vary from 20 days at 30 °C to 56 days at 17 °C and the lowest thermal threshold is estimated at 10.2 °C. The optimal temperature for immature development is 32.5 °C. Total fecundity (eggs per female) ranges from 105.3 (at 21 °C) to 41 (at 35 °C). The longevity decreases with temperature increase. The intrinsic rate of increase ranges from 0.0450 (at 17 °C) to 0.123 (at 30 °C). The functional relationships between temperature and life-history parameters are used to evaluate the effect of temperature on the population dynamics. Such mathematical relationships could provide a basis for future development of population models.     

Draft Genome Sequence of Rickettsia sp. Strain MEAM1, Isolated from the Whitefly Bemisia tabaci

We report the draft genome sequence of the Rickettsia sp. strain MEAM1, which is a facultative symbiont from an invasive species of the whitefly Bemisia tabaci. The total length of the assembled genome is approximately 1.24 Mb, with 335 scaffolds and 1,247 coding sequences predicted within the genome.     

Genetic relationships among biotypes of Bemisia tabaci (Hemiptera: Aleyrodidae) based on AFLP analysis

Genetic similarities between 13 samples belonging to nine reference biotypes and two field populations of Bemisia tabaci (Gennadius), one field population of B. medinae Gómez-Menor and another of B. afer Priesner & Hosny, were evaluated using amplified fragment length polymorphism (AFLP) markers. The results indicate that B. tabaci biotypes can be grouped together with a minimum similarity coefficient of 0.32 and separated from the two other species with a similarity coefficient of 0.07. Bemisia tabaci biotypes were grouped in four clusters which comprised: (i) Near East and Indian subcontinent biotypes; (ii) B and Q biotypes plus a Nigerian population from cowpea; (iii) New World A biotype; and (iv) S biotype and a Nigerian population from cassava. These results were consistent with a previous grouping of biotypes based on RAPD-PCR analysis. The AFLP assay allowed the scoring of a total of 354 polymorphic bands in two reaction events with the use of two primer combinations.     

Host plant influence on susceptibility of Bemisia tabaci (Hemiptera: Aleyrodidae) to insecticides

A resistance monitoring program conducted for the polyphagous whitefly, Bemisia tabaci (Gennadius), in Imperial Valley, CA, USA generated a large set of LC50s for adults collected from broccoli, cantaloupe and cotton crops over a four-year period. A vial bioassay and, subsequently, a yellow-sticky card bioassay produced similar temporal profiles of relative susceptibilities to the pyrethroid insecticide bifenthrin. Both bioassays revealed that whiteflies collected from broccoli were significantly less susceptible to bifenthrin compared to the other two crops. A similar finding was observed for endosulfan and the mixture of bifenthrin+endosulfan in the yellow-sticky card bioassay. The possibility that seasonal differences contributed to the observed differences in susceptibility provided the impetus to conduct a reciprocal transfer experiment using broccoli (or kale) and cantaloupe grown simultaneously in the field and greenhouse. Whitefly adults collected from an organic farm over three consecutive weeks had significantly higher LC50s on kale than those collected the same day on cantaloupe. After culturing in the greenhouse on broccoli or cantaloupe and testing again, LC50s remained significantly higher on broccoli after one week and again at the F1 generation. In contrast, whiteflies originating on kale in the field and transferred to cantaloupes in the greenhouse had significantly reduced LC50s at the F1 generation. When tested against the bifenthrin+endosulfan mixture, significantly higher LC50s were generated for whiteflies reared on broccoli in the greenhouse at one week and the F1 compared to the field source from cantaloupes. The consistently higher LC50s for whiteflies on broccoli and other Brassica spp. crops, compared to cantaloupes or cotton, point to statistically significant host-plant influences that are expressed in both field-collected and greenhouse-reared populations of whiteflies.     

应用RNA干扰技术调节大鼠c-jun基因的表达

目的:应用RNA干扰(RNAi)技术调节大鼠c-jun基因在Cos-7细胞中的表达.方法:分别构建大鼠c-jun基因的RNA干扰载体和真核表达GFP(绿色荧光蛋白)载体,将两者共转染Cos-7细胞,镜下观察大鼠C-Jun-GFP融合蛋白的表达,应用Western blot方法检测抑制效率.结果:酶切和测序结果表明,大鼠c-jun基因的RNA干扰载体和真核表达GFP载体构建成功,镜下及Western blot共转染结果均显示随着RNA干扰载体浓度的增加C-Jun-GFP融合蛋白表达量逐渐减少.结论:在Cos-7细胞中应用RNAi技术成功调节大鼠c-jun基因的表达.     

Field evaluation of resistance to pyriproxyfen in Bemisia tabaci (B biotype)

We determined effects of aerial sprays of the insect growth regulator pyriproxyfen on sweetpotato whitefly, Bemisia tabaci (Gennadius) (B biotype), in Arizona cotton (Gossypium spp.) fields. We measured survival for males and females from a susceptible strain and a laboratory-selected resistant strain, as well as for hybrid female progeny from crosses between the strains. Insects were exposed directly to pyriproxyfen sprays in the field or indirectly in the laboratory by rearing them on sprayed leaves collected from the field. In all tests, survival was higher for the resistant strain than the susceptible strain, but did not differ between sexes in each strain. Survival to the adult stage did not differ between eggs and nymphs directly exposed to sprays. For susceptible and hybrid individuals, survival was lower on leaves collected the day of spraying than on leaves collected 2 wk after spraying. In contrast, survival of resistant individuals did not differ based on the timing of exposure. Dominance of resistance to pyriproxyfen depended on the type of exposure. Resistance was partially or completely dominant in direct exposure bioassays and on leaves collected 2 wk after spraying (h > 0.6). Resistance was partially recessive on leaves collected the day of spraying (mean h = 0.34). Rapid evolution of resistance to pyriproxyfen could occur if individuals in field populations with traits similar to those of the laboratory-selected strain examined here were treated intensively with this insecticide.     

Assessment of cross-resistance potential to neonicotinoid insecticides in Bemisia tabaci (Hemiptera: Aleyrodidae)

Laboratory bioassays were carried out with four neonicotinoid insecticides on multiple strains of Bemisia tabaci (Gennadius) to evaluate resistance and cross-resistance patterns. Three imidacloprid-resistant strains and field populations from three different locations in the southwestern USA were compared in systemic uptake bioassays with acetamiprid, dinotefuran, imidacloprid and thiamethoxam. An imidacloprid-resistant strain (IM-R) with 120-fold resistance originally collected from Imperial Valley, California, did not show cross-resistance to acetamiprid, dinotefuran or thiamethoxam. The Guatemala-resistant strain (GU-R) that was also highly resistant to imidacloprid (RR=109-fold) showed low levels of cross-resistance when bioassayed with acetamiprid and thiamethoxam. However, dinotefuran was more toxic than either imidacloprid or thiamethoxam to both IM-R and GU-R strains as indicated by low LC50s. By contrast, a Q-biotype Spanish-resistant strain (SQ-R) of B. tabaci highly resistant to imidacloprid demonstrated high cross-resistance to the two related neonicotinoids. Field populations from Imperial Valley (California), Maricopa and Yuma (Arizona), showed variable susceptibility to imidacloprid (LC50s ranging from 3.39 to 115 microg ml(-1)) but did not exhibit cross-resistance to the three neonicotinoids suggesting that all three compounds would be effective in managing whiteflies. Yuma populations were the most susceptible to imidacloprid. Dinotefuran was the most toxic of the four neonicotinoids against field populations. Although differences in binding at the target site and metabolic pathways may influence the variability in cross-resistance patterns among whitefly populations, comparison of whitefly responses from various geographic regions to the four neonicotinoids indicates the importance of ecological and operational factors on development of cross-resistance to the neonicotinoids.