Patterns of host cell inheritance in the bacterial symbiosis of whiteflies

Whiteflies possess bacterial symbionts Candidatus Portiera aleyrodidium that are housed in specialized cells called bacteriocytes and are faithfully transmitted via the ovary to insect offspring. In one whitefly species studied previously, Bemisia tabaci MEAM1, transmission is mediated by somatic inheritance of bacteriocytes, with a single bacteriocyte transferred to each oocyte and persisting through embryogenesis to the next generation. Here, we investigate the mode of bacteriocyte transmission in two whitefly species, B. tabaci MED, the sister species of MEAM1, and the phylogenetically distant species Trialeurodes vaporariorum. Microsatellite analysis supported by microscopical studies demonstrates that B. tabaci MED bacteriocytes are genetically different from other somatic cells and persist through embryogenesis, as for MEAM1, but T. vaporariorum bacteriocytes are genetically identical to other somatic cells of the insect, likely mediated by the degradation of maternal bacteriocytes in the embryo. These two alternative modes of transmission provide a first demonstration among insect symbioses that the cellular processes underlying vertical transmission of bacterial symbionts can diversify among related host species associated with a single lineage of symbiotic bacteria.     

Cover Caption

The whitefly Bemisia tabaci contains many big bacteriocytes, which house the primary symbiont “Candidatus Portiera aleyrodidarum” and a variety of secondary symbionts (see pages 194–206). The primary symbiont provides essential amino acids to hosts and the secondary symbionts can affect the life parameters of B. tabaci including fecundity, development time and sex‐ratio. In addition, the secondary symbionts of B. tabaci have been reported to confer resistance to natural enemies, heat stress and insecticides. Photo in cover shows a nymph of the whitefly and the two kidney‐like organs are the assemblage of bacteriocytes. Image by Xiao‐Li Bing.     

Differential responses between a vector species Bemisia tabaci and a nonvector species Trialeurodes vaporariorum following ingestion of tomato yellow leaf curl virus

In transmitting plant viruses, insect vectors undergo physiological and behavioral alterations. The whitefly Bemisia tabaci is a vector of tomato yellow leaf curl virus (TYLCV), causing severe damages to various horticultural crop plants. To determine whether whitefly alteration is specific to vector species, the responses to TYLCV ingestion were compared between B. tabaci and Trialeurodes vaporariorum, a nonvector for TYLCV. The two species were reared on TYLCV‐infected and noninfected tomato, a host of TYLCV, and their longevity and fecundity were determined while rearing in either tomato or eggplant, a nonhost of TYLCV. TYLCV‐ingested B. tabaci increased their developmental rates but reduced fecundity when they were reared in either tomato or eggplant compared with those of TYLCV‐free ones. In contrast, TYLCV‐ingested T. vaporariorum did not show any of the aforementioned changes when reared on both plant species. In addition, TYLCV‐ingested B. tabaci increased their levels of three heat shock protein genes ( hsp20, hsp70, and hsp90) against thermal stress, whereas TYLCV‐ingested T. vaporariorum did not. The presence of TYLCV virions was identified in two colonies of both species via polymerase chain reaction analysis. TYLCV was detected in the whole body, saliva, and eggs of B. tabaci, while TYLCV was detected only in the whole body but not in the saliva and eggs of T. vaporariorum. The present results strongly indicated that TYLCV specifically manipulate physiological processes of the vector species, B. tabaci.     

Cloning and expression of heat shock protein genes in two whitefly species in response to thermal stress

Two whitefly species, Trialeurodes vaporariorum and Bemisia tabaci biotype B were shown to have different temperature tolerance and seasonal dynamics. To determine whether this variation in thermal tolerance is related to different expression patterns of heat shock protein (hsp) genes during temperature stress, we obtained complete cDNA sequences for hsp90, hsp70 and hsp20, and analysed their expression profiles across temperature gradients by real‐time quantitative polymerase chain reaction (PCR). Six full‐length cDNAs were cloned and sequenced from these two species. The full‐length cDNAs of hsp90s contain 2166 and 2157 bp open‐reading frames (ORF) which encode proteins with calculated molecular weights of 83 013 and 82 857 Da in T. vaporariorum and B. tabaci, respectively. The 1947 and 1959 bp ORFs of whitefly hsp70s comprise 649 and 653 amino acids with the calculated masses of 70 885 and 71 008 Da in T. vaporariorum and B. tabaci, respectively. Both complete cDNAs of hsp20 of T. vaporariorum and B. tabaci contain 585 bp ORFs and deduced amino acid sequences had molecular weights of 21 559 and 21 539 Da, respectively. The hsp expression profile results showed that temperatures for onset (T_on) or maximal (T_max) induction of hsp expression in T. vaporariorum were generally 2–6°C lower than those in B. tabaci. These results suggest that the T_on (or T_max) of hsps can represent the differences in temperature tolerance of these two whitefly species, and may be used to determine their natural geographical distribution and natural population seasonal dynamics. Significant upregulation of most hsps were observed when temperature stress was lifted, except that hsp70 and hsp20 of B. tabaci did not respond to the cold stress, indicating that response to heat and cold stress may have a different genetic and physiological basis in two whitefly species. These results highlight the importance of understanding the complexity of the heat shock response across multiple isoforms while attempting to link them to whole‐organism traits such as thermal tolerance.     

Fitness of Encarsia sophia (Hymenoptera: Aphelinidae) parasitizing Trialeurodes vaporariorum and Bemisia tabaci (Hemiptera: Aleyrodidae)

Abstract Fitness and efficacy of Encarsia sophia (Girault & Dodd) (Hymenoptera: Aphelinidae) as a biological control agent was compared on two species of whitefly (Hemiptera: Aleyrodidae) hosts, the relatively smaller sweetpotato whitefly, Bemisia tabaci (Gennadius) biotype ‘B’, and the larger greenhouse whitefly, Trialeurodes vaporariorum (Westwood). Significant differences were observed on green bean (Phaseolus vulgaris L.) in the laboratory at 27 ± 2°C, 55%± 5% RH, and a photoperiod of 14: 10 h (L: D). Adult parasitoids emerging from T. vaporariorum were larger than those emerging from B. tabaci, and almost all biological parameters of E. sophia parasitizing the larger host species were superior except for the developmental times of the parasitoids that were similar when parasitizing the two host species. Furthermore, parasitoids emerging from T. vaporariorum parasitized more of these hosts than did parasitoids emerging from B. tabaci. We conclude that E. sophia reared from larger hosts had better fitness than from smaller hosts. Those from either host also preferred the larger host for oviposition but were just as effective on smaller hosts. Therefore, larger hosts tended to produce better parasitoids than smaller hosts.     

Genetic diversity and geographic distribution of Bemisia tabaci and Trialeurodes vaporariorum in Costa Rica

The tobacco whitefly Bemisia tabaci (Gennadius) cryptic species complex and of the greenhouse whitefly Trialeurodes vaporariorum (Westwood) are extensively reported as destructive pests in vegetable crops worldwide. A survey was conducted in 2011 and 2012 to determine the occurrence and genetic diversity present in the populations of these whiteflies in the major vegetable production areas of Costa Rica. Insect samples were collected from sweet pepper (Capsicum annuum L.), tomato (Solanum lycopersicum L.), common bean (Phaseolus vulgaris L.) and weeds present in commercial crops either in open field or greenhouse conditions. PCR‐RFLP analysis of mitochondrial cytochrome c oxidase subunit 1 gene (mtCOI) sequences of 621 whitefly individuals confirmed the presence of the Mediterranean (MED) type of the B. tabaci and of T. vaporariorum in most sampled regions. Also, individuals of the Middle East‐Asia Minor 1 (MEAM1) type of the B. tabaci were observed in low numbers. Contingency analyses based on type of crop, geographical region, whitefly species, year of collection and production system confirmed that T. vaporariorum was the most frequent species in vegetable production areas in Costa Rica, both in greenhouses and in open fields. B. tabaci MED is likely spreading to new areas of the country, whereas B. tabaci MEAM1 was mostly absent or rarely found. Comparisons of mtCOI sequences from B. tabaci individuals revealed the presence of four B. tabaci sequence haplotypes (named MED‐i, MED‐ii, MEAM1‐i, MEAM1‐xviii) in Costa Rica, three of them identical to B. tabaci haplotypes previously reported in the Western Hemisphere and other parts of the world. Analysis of sequences of T. vaporariorum individuals revealed a more complex population with the presence of 11 haplotypes, two of which were identical to T. vaporariorum sequences reported from other countries.     

Variation between laboratory populations in the performance of the parasitoid Encarsia formosa on two host species, Bemisia tabaci and Trialeurodes vaporariorum

We tested the hypothesis that populations of the parthenogenetic parasitic wasp Encarsia formosa Gahan (Hymenoptera: Aphelinidae) differed in their ability to use two different host species, Bemisia tabaci Gennadius (Homoptera: Aleyrodidae) and Trialeurodes vaporariorum Westwood (Homoptera: Aleyrodidae). Of the three wasp populations tested, two populations had been reared for many generations on B. tabaci and one population had been reared for many years on T. vaporariorum. Performance was measured by the number of whitefly nymphs that were successfully parasitized by individual wasps, and performance on either host was measured in separate experiments. There was variation between wasp populations in their performance on the host B. tabaci, with one wasp population reared for many years on this host performing considerably better than the other two populations. There were no significant differences between populations in their use of the preferred host, T. vaporariorum. The experiments were conducted in such a way that we could distinguish heritable differences between populations from environmentally-induced conditioning differences due to the immediate host from which an individual wasp enclosed. In either experiment there were no significant effects of conditioning, although there was a trend within each population for wasps conditioned on T. vaporariorum to have higher performance than those conditioned on B. tabaci. Thirdly, we conducted a selection experiment, initiated with wasps from a single population historically reared on T. vaporariorum, to measure the effect of laboratory rearing on different hosts for 17 generations. We did not see any difference in the performance of wasps on B. tabaci after this period of rearing on either of the two hosts. In summary, populations of E. formosa do differ in their relative performance on B. tabaci. The one population that was tested further did not show any response to selection by rearing, but the ability to respond to selection on performance may not be equal for all populations. The possibility that wasp populations have differential performance on particular hosts may affect the use of this species as a biological control agent.     

Host preferences and biotic potential of <Emphasis Type="Italic">Trialeurodes vaporariorum</Emphasis> and <Emphasis Type="Italic">Bemisia tabaci</Emphasis> (Hemiptera: Aleyrodidae) in tomato and pepper

Whiteflies Bemisia tabaci (Gennadius) and Trialeurodes vaporariorum (Westwood) (Hemiptera: Aleyrodidae) are important pests in pepper (Capsicum annuum L.) and tomato (Solanum lycopersicum L.) crops in many countries. Contrary to what is observed for all other countries, in Uruguay, B. tabaci is mainly found on pepper and rarely on tomato, while T. vaporariorum is exclusively found on tomato. This study tested the oviposition preferences and biotic potential of these two whiteflies reared on both host plants. The developmental time, survival rates, longevity, fecundity and main population parameters were characterized. Both whitefly species showed different preference patterns regarding their host plants. T. vaporariorum preferred tomato instead of pepper to oviposit. Their developmental time is longer on pepper. B. tabaci preferred pepper, but the difference from tomato was not very strong. Pepper affects the biotic expression of T. vaporariorum negatively, while B. tabaci is able to develop equally on both host plants. These results show that the distribution differences of both whiteflies observed on both host plants could have a biological basis.     

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。     

Intracellular symbionts drive sex ratio in the whitefly by facilitating fertilization and provisioning of B vitamins

Symbionts can regulate animal reproduction in multiple ways, but the underlying physiological and biochemical mechanisms remain largely unknown. The presence of multiple lineages of maternally inherited, intracellular symbionts (the primary and secondary symbionts) in terrestrial arthropods is widespread in nature. However, the biological, metabolic, and evolutionary role of co-resident secondary symbionts for hosts is poorly understood. The bacterial symbionts Hamiltonella and Arsenophonus have very high prevalence in two globally important pests, the whiteflies Bemisia tabaci and Trialeurodes vaporariorum, respectively. Both symbionts coexist with the primary symbiont Portiera in the same host cell (bacteriocyte) and are maternally transmitted. We found that elimination of both Hamiltonella and Arsenophonous by antibiotic treatment reduced the percentage of female offspring in whiteflies. Microsatellite genotyping and cytogenetic analysis revealed that symbiont deficiency inhibited fertilization in whiteflies, leading to more haploid males with one maternal allele, which is consistent with distorted sex ratio in whiteflies. Quantification of essential amino acids and B vitamins in whiteflies indicated that symbiont deficiency reduced B vitamin levels, and dietary B vitamin supplementation rescued fitness of whiteflies. This study, for the first time, conclusively demonstrates that these two intracellular symbionts affect sex ratios in their whitefly hosts by regulating fertilization and supplying B vitamins. Our results reveal that both symbionts have the convergent function of regulating reproduction in phylogenetically-distant whitefly species. The 100% frequency, the inability of whiteflies to develop normally without their symbiont, and rescue with B vitamins suggests that both symbionts may be better considered co-primary symbionts.Subject terms: Microbial ecology, Applied microbiology     

Effect of the primary host for production of both sexes on the mating interaction in an autoparasitoid species

Encarsia sophia (Girault and Dodd) is an autoparasitoid in the hymenopteran family Aphelinidae. The females develop as primary parasitoids on whitefly nymphs (primary hosts), whereas the males develop as hyperparasitoids on their own species or on other primary parasitoid species (secondary hosts). The autoparasitoids not only parasitise whiteflies but also kill them with strong host-feeding capacity. In this study, female and male E. sophia were reared on the primary hosts Trialeurodes vaporariorum and Bemisia tabaci ‘Q’, and the host-feeding and parasitism of wasps on both whitefly species were determined for the four possible different mating combinations: (i) E. sophia females reared on B. tabaci (ESF-BT) mated with E. sophia males from B. tabaci (ESM-BT), (ii) E. sophia females reared on T. vaporariorum (ESF-TV) mated with E. sophia males from T. vaporariorum (ESM-TV), (iii) ESF-BT mated with ESM-TV, and (iv) ESF-TV mated with ESM-BT. ESF-TV mated with ESM-TV killed the largest percentage of whitefly nymphs through host feeding. The ESF-TV with larger body size mating with larger ESM-TV killed more whitefly nymphs through host feeding than those mating with smaller ESM-BT. Whether B. tabaci or T. vaporariorum were used as hosts, ESF-TV mated with ESM-TV and ESM-BT and ESF-BT mated with ESM-BT significantly parasitised more whitefly nymphs than ESF-BT mated with ESM-TV. In general, ESF-BT mated with ESM-TV killed significantly fewer whitefly nymphs through parasitism and host feeding than the other three mating combinations on both whitefly species. These results indicated that the performance of autoparasitoids on insect pests was not only dependent on females but was also affected by mating with males from different primary host species.     

Temporal changes of symbiont density and host fitness after rifampicin treatment in a whitefly of the Bemisia tabaci species complex

Microbial symbionts are essential or important partners to phloem‐feeding insects. Antibiotics have been used to selectively eliminate symbionts from their host insects and establish host lines with or without certain symbionts for investigating functions of the symbionts. In this study, using the antibiotic rifampicin we attempted to selectively eliminate certain symbionts from a population of the Middle East‐Asia Minor 1 whitefly of the Bemisia tabaci species complex, which harbors the primary symbiont “Candidatus Portiera aleyrodidarum” and two secondary symbionts “Candidatus Hamiltonella defensa” and Rickettsia. Neither the primary nor the secondary symbionts were completely depleted in the adults (F0) that fed for 48 h on a diet treated with rifampicin at concentrations of 1–100 μg/mL. However, both the primary and secondary symbionts were nearly completely depleted in the offspring (F1) of the rifampicin‐treated adults. Although the F1 adults produced some eggs (F2), most of the eggs failed to hatch and none of them reached the second instar, and consequently the rifampicin‐treated whitefly colony vanished at the F2 generation. Interestingly, quantitative polymerase chain reaction assays showed that in the rifampicin‐treated whiteflies, the density of the primary symbiont was reduced at an obviously slower pace than the secondary symbionts. Mating experiments between rifampicin‐treated and untreated adults demonstrated that the negative effects of rifampicin on host fitness were expressed when the females were treated by the antibiotic, and whether males were treated or not by the antibiotic had little contribution to the negative effects. These observations indicate that with this whitefly population it is not feasible to selectively eliminate the secondary symbionts using rifampicin without affecting the primary symbiont and establish host lines for experimental studies. However, the extinction of the whitefly colony at the second generation after rifampicin treatment indicates the potential of the antibiotic as a control agent of the whitefly pest.     

First interception of Bemisia tabaci Mediterranean (Q biotype) in Serbia

The whitefly Bemisia tabaci (Gennadius) is a cosmopolitan insect pest and causes great damage to many agricultural crops by direct feeding, and transmitting many plant viruses causing disastrous effects. The global distribution of B. tabaci was previously investigated, but so far, it has not been confirmed in Serbia. Sampling conducted on plants at three localities in Serbia revealed the presence of three whitefly species, one is B. tabaci Mediterranean (MED) species, a quarantine pest. Additionally, Trialeurodes vaporariorum Westwood and an Aleurodes sp. that was not identified to the species level were also confirmed. The last two are common and widespread species in Serbia. Whitefly collections were made from cabbage, kale and oilseed rape plants (Brassica oleracea, B. oleracea var. sabauda, B. napus, respectively) grown in the field, on sunflower (Helianthus annuus) grown in the greenhouse and on potted hibiscus (Hibiscus sp.) and poinsettia (Euphorbia pulcherrima) plants grown indoors. DNA sequence analysis from a portion of the Cytochrome Oxidase I gene (COI) revealed the presence of B. tabaci MED species, which is considered to be a very important pest worldwide and has been reported from neighbouring countries, however, not yet from Serbia. Bemisia tabaci MED is recorded here for the first time in Serbia. This first interception suggests its introduction with ornamentals grown indoors, with no outdoor reports and that it is not yet spread. This report is an important alert for local authorities to take the necessary steps in monitoring and preventing its possible expansion.     

Ultrastructure of the endosymbionts of the whitefly,Bemisia tabaci andTrialeurodes vaporariorum

Summary The ultrastructure of the mycetocytes and mycetome micro-organisms of the sweetpotato whitefly,Bemisia tabaci Genn. andTrialeurodes vaporariorum West are described. InB. tabaci, two morphologically distinct types of micro-organisms were observed in mycetocytes. The predominant type lacked a distinct cell wall, was pleomorphic in shape with a surrounding vacuole. The second type was a coccoid organism, with inner and outer cell membranes. The coccoid organism was often found in groups of varying number within vacuoles, and in many cases appeared to be undergoing degradation. InT. vaporariorum mycetocytes, pleomorphic and coccoid organisms were found, although the coccoid micro-organism inT. vaporariorum, had a thicker cell wall than the coccoid micro-organism inB. tabaci.Abbreviations C coccoid micro-organism - P pleomorphic micro-organism     

The status ofBemisia tabaci (Hom.: Aleyrodidae),Trialeurodes vaporariorum (Hom.: Aleyrodidae) and their natural enemies in Crete

The non «B» biotype ofBemisia tabaci (Gennadius) is recorded for the first time in Crete in 1992, in the north east and south east of the island.Trialeurodes vaporariorum (Westwood) is the predominant whitefly on plants in the north and west of the island. Three surveys of Crete were made in 1992 and 1993 for natural enemies ofB. tabaci andT. vaporariorum and resulted in the collection of 4 species ofEncarsia, (plus a number of species that are unidentifiable at this time), anEretmocerus sp. (unidentifiable at this time) and a fungal pathogen,Paecilomyces farinosus (Dickson Ex Fries) Brown &; Smith.Encarsia adrianae was identified fromT. vaporariorum; which constitutes its most westerly distribution point and a new host record respectively.B. tabaci andT. vaporariorum were found on horticultural crops, ornamentals and weeds. Populations of both whitefly species were severely depleted on field hosts throughout the island during the winter of 1992/93. Climatic constraints, competition withT. vaporariorum in otherwise suitable niches, effective natural enemies and an observed low level of polyphagy may explain the present limited distribution of the non «B» biotype ofB. tabaci in Crete.     

Abundance of Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae) and its parasitoids on vegetables and cassava plants in Burkina Faso (West Africa)

The whitefly Bemisia tabaci is a pest of many agricultural and ornamental crops worldwide and particularly in Africa. It is a complex of cryptic species, which is extremely polyphagous with hundreds of host plants identified around the world. Previous surveys in western Africa indicated the presence of two biotypes of the invasive MED species (MED‐Q1 and MED‐Q3) living in sympatry with the African species SSA and ASL. This situation constitutes one of the rare cases of local coexistence of various genetic entities within the B. tabaci complex. In order to study the dynamics of the distribution and abundance of genetic entities within this community and to identify potential factors that could contribute to coexistence, we sampled B. tabaci populations in Burkina Faso in 2015 and 2016 on various plants, and also their parasitoids. All four genetic entities were still recorded, indicating no exclusion of local species by the MED species. While B. tabaci individuals were found on 55 plant species belonging to eighteen (18) families showing the high polyphagy of this pest, some species/biotypes exhibited higher specificity. Two parasitoid species (Eretmocerus mundus and Encarsia vandrieschei) were also recorded with E. mundus being predominant in most localities and on most plants. Our data indicated that whitefly abundance, diversity, and rate of parasitism varied according to areas, plants, and years, but that parasitism rate was globally highly correlated with whitefly abundance suggesting density dependence. Our results also suggest dynamic variation in the local diversity of B. tabaci species/biotypes from 1 year to the other, specifically with MED‐Q1 and ASL species. This work provides relevant information on the nature of plant–B. tabaci‐parasitoid interactions in West Africa and identifies that coexistence might be stabilized by niche differentiation for some genetic entities. However, MED‐Q1 and ASL show extensive niche overlap, which could ultimately lead to competitive exclusion.     

Inactivation of Wolbachia Reveals Its Biological Roles in Whitefly Host

Background

The whitefly Bemisia tabaci is cryptic species complex composed of numerous species. Individual species from the complex harbor a diversity of bacterial endosymbionts including Wolbachia. However, while Wolbachia is known to have a number of different roles, its role in B. tabaci is unclear. Here, the antibiotic rifampicin is used to selectively eliminate Wolbachia from B. tabaci so as to enable its roles in whitefly development and reproduction to be explored. The indirect effects of Wolbachia elimination on the biology of Encarsia bimaculata, a dominant parasitoid of B. tabaci in South China, were also investigated.

Methodology/Principal Finding

qRT-PCR and FISH were used to show that after 48 h exposure to 1.0 mg/ml rifampicin, Wolbachia was completely inactivated from B. tabaci Mediterranean (MED) without any significant impact on either the primary symbiont, Portiera aleyrodidarum or any of the other secondary endosymbionts present. For B. tabaci MED, Wolbachia was shown to be associated with decreased juvenile development time, increased likelihood that nymphs completed development, increased adult life span and increased percentage of female progeny. Inactivation was associated with a significant decrease in the body size of the 4^th instar which leads us to speculate as to whether Wolbachia may have a nutrient supplementation role. The reduction in nymph body size has consequences for its parasitoid, E. bimaculata. The elimination of Wolbachia lead to a marked increase in the proportion of parasitoid eggs that completed their development, but the reduced size of the whitefly host was also associated with a significant reduction in the size of the emerging parasitoid adult and this was in turn associated with a marked reduction in adult parasitoid longevity.

Conclusions/Significance

Wolbachia increases the fitness of the whitefly host and provides some protection against parasitization. These observations add to our understanding of the roles played by bacterial endosymbionts.     

Selection of Beauveria bassiana isolates for control of the whiteflies Bemisia tabaci and Trialeurodes vaporariorum on the basis of their virulence, thermal requirements, and toxicogenic activity

As part of a 3-fold approach to select potential mycoinsecticides for whitefly control, we evaluated infectivity, thermal requirements, and toxicogenic activity of the entomopathogenic fungus Beauveria bassiana (Ascomycota: Clavicipitaceae) under laboratory conditions. Twenty-five native B. bassiana isolates and a commercially available mycoinsecticide (based on B. bassiana) were evaluated for virulence to fourth instar nymphs of sweetpotato whitefly, Bemisia tabaci, and greenhouse whitefly, Trialeurodes vaporariorum, at a concentration of 1 × 10⁷ conidia/ml. All isolates were pathogenic for both whitefly species, whereas mortality rates varied from 3 to 85%. A second series of bioassays was conducted on 10 selected isolates using four 10-fold concentrations ranging from 1 × 10⁵ to 1 × 10⁸ conidia/ml. Median lethal concentrations (LC₅₀) of the four most virulent isolates varied from 1.1 × 10⁵ to 6.2 × 10⁶ conidia/ml and average survival time (AST) of treated nymphs from 5.9 to 7.4 days. T. vaporariorum were significantly more susceptible to all B. bassiana isolates than B. tabaci. The thermal biology of the eight most virulent isolates to both whitefly species was investigated at six temperatures (10–35 °C). The colony radial growth rate was estimated from the slope of the linear regression of colony radius on time and data were then fitted to a modified generalized β function that accounted for 90.5–99.3% of the data variance. Optimum temperatures for extension rate ranged from 23.1 to 27.1 °C, whereas maximum temperatures for fungal growth varied from 31.8 to 36.6 °C. On the basis of their virulence and thermal requirements, three isolates showed promise as candidates for whitefly management in Mediterranean greenhouses. Whilst in vitro production of macromolecular compounds toxic to Galleria mellonella larvae was not a requisite for virulence, ASTs of larvae injected with Sephadex G-25 fractions from candidate isolates ranged from 1.4 to 3.7 days compared with 5–6 days for non-toxic G-25 fractions. In addition, proteinase K treatment significantly reduced their toxic activity suggesting that they were proteins and revealing the potential of these isolates to be further improved through biotechnology to kill the pest more quickly.