PSR (paternal sex ratio) chromosomes: the ultimate selfish genetic elements

PSR (paternal sex ratio) chromosomes are a type of supernumerary (or B) chromosomes that occur in haplodiploid arthropods. They are transmitted through sperm but then cause loss of the paternal chromosomes (except themselves) early in development. As a result, PSR chromosomes convert diploid fertilized eggs (which would normally develop into females) into haploid males that carry a PSR chromosome. Because they act by completely eliminating the haploid genome of their hosts, PSR chromosomes are the most extreme form of selfish or parasitic DNA known. PSR was originally described in the parasitic wasp Nasonia vitripennis (Pteromalidae). A second PSR chromosome has been found in Trichogramma kaykai, an egg parasitoid from a different family of Hymenoptera (Trichogrammatidae). We argue that PSR chromosomes are likely to be widespread in haplodiploid organisms, but have so far gone under reported due to a paucity of population genetic studies in haplodiploids. We describe the two known PSR systems and related phenomena, and models indicating the conditions conducive to increase of PSR like chromosomes in haplodiploids.     

Effect of fertilized,unfertilized, and UV‐irradiated hosts on parasitism and suitability for Trichogramma parasitoids

Trichogramma spp. (Hymenoptera: Trichogrammatidae) parasitoids have been commonly used as biological control agents in insect pest management. Host quality is believed to influence parasitism, host preference, and suitability for parasitoids. To date, limited studies have compared the parasitism of Trichogramma parasitoids on fertilized, unfertilized, and sterilized host eggs. Hence, we studied the performance of three Trichogramma egg parasitoids, Trichogramma japonicum Ashmead, Trichogramma chilonis Ishii, and Trichogramma leucaniae Pang & Chen, on fertilized, unfertilized, and ultraviolet (UV)‐irradiated fertilized (UVF) eggs of rice moth, Corcyra cephalonica (Stainton) (Lepidoptera: Pyralidae). In a no‐choice test, T. japonicum and T. leucaniae parasitized significantly more fertilized or UVF than unfertilized hosts, and T. chilonis parasitized significantly more UVF than either fertilized or unfertilized hosts. In a choice test, all three Trichogramma parasitoids parasitized UVF hosts the most, and unfertilized hosts the least. There were similar percentages of adult emergence and female progeny among fertilized, unfertilized, and UVF hosts for all three Trichogramma parasitoids, except that T. japonicum had significantly lower adult emergence on fertilized hosts. We also found that all three Trichogramma parasitoids developed slower on unfertilized hosts. Regardless of host treatments, T. leucaniae had the longest developmental time and T. chilonis had the shortest. We conclude that Trichogramma parasitoids prefer parasitizing UVF eggs of C. cephalonica without negative effects on their emergence and sex allocation.     

The origin of a selfish B chromosome triggering paternal sex ratio in the parasitoid wasp Trichogramma kaykai

This study uses molecular and cytogenetic methods to determine the origin of a B chromosome in some males of the wasp Trichogramma kaykai. This so-called paternal sex ratio (PSR) chromosome transmits only through sperm and shortly after fertilization triggers degeneration of the paternal genome, while keeping itself intact. The resulting embryos develop into haploid B-chromosome-carrying males. Another PSR chromosome with a very similar mode of action is found in the distantly related wasp Nasonia vitripennis and its origin was traced by transposon similarity to the genus Trichomalopsis, which is closely related to Nasonia. To determine whether both PSR chromosomes have a similar origin we aimed to reveal the origin of the Trichogramma PSR chromosome. Using fluorescent in situ hybridization, we discovered a major satellite repeat on the PSR chromosome, the 45S ribosomal DNA. Analysis of the internal transcribed spacer 2 (ITS2) of this repeat showed the presence of multiple ITS2 sequences on the PSR chromosome resembling either the ITS2 of T. oleae or of T. kaykai. We therefore conclude that the Trichogramma PSR chromosome originates from T. oleae or a T. oleae-like species. Our results are consistent with different origins for the PSR chromosomes in Trichogramma and Nasonia.     

The relationship between egg load and fecundity among Trichogramma parasitoids

1. The relationships between parasitoid egg load, size, and age (3–72 h) for Trichogramma minutum, T. platneri, and T. pretiosum, reared from two factitious hosts, Ephestia kuehniella and Sitotroga cerealella, were evaluated to test the hypothesis that 24‐h egg load can be used to estimate the fecundity of Trichogramma parasitoids. 2. Egg load increased in relation to female age over the first 3 days of adult life for all three Trichogramma species to a mean egg storage capacity of 46.7 eggs for T. minutum, 41.1 for T. pretiosum, and 35.7 for T. platneri. At 24 h of age, T. minutum had matured enough eggs to fill 67% of its storage capacity, in comparison with 74% for T. pretiosum and 91% for T. platneri. There was a positive relationship between egg load and parasitoid size for all ages of the three Trichogramma species reared from both hosts (with the exception of T. platneri at 3 h post emergence), accounting for 14–69% of the variance in egg load. 3. The potential fecundity, realised (3 day cumulative) fecundity, and oviposition rate (potential fecundity/longevity) of T. platneri were all related linearly to size‐dependent variation in 24‐h egg load, but only the realised fecundity of T. pretiosum, and none of the reproductive characteristics of T. minutum. It is suggested that 24‐h egg load may not be an accurate measure of egg storage capacity in parasitoids and should be used cautiously to represent fecundity. 4. The potential fecundity of seven Trichogramma species reared from E. kuehniella varied from 55 to 150, but neither potential fecundity nor oviposition rate was related significantly to egg load (represented by eggs laid during first 24 h). Selection to avoid egg depletion in the attack of gregarious hosts appears most likely to account for the variation in potential fecundity among Trichogramma species.     

Induction of paternal genome loss by the paternal-sex-ratio chromosome and cytoplasmic incompatibility bacteria (Wolbachia): A comparative study of early embryonic events

Paternal genome loss (PGL) during early embryogenesis is caused by two different genetic elements in the parasitoid wasp, Nasonia vitripennis. Paternal sex ratio (PSR) is a paternally inherited supernumerary chromosome that disrupts condensation of the paternal chromosomes by the first mitotic division of fertilized eggs. Bacteria belonging to the genus Wolbachia are present in Nasonia eggs and also disrupt paternal chromosome condensation in crosses between cytoplasmically incompatible strains. Cytoplasmic incompatibility Wolbachia are widespread in insects, whereas PSR is specific to this wasp. PGL results in production of male progeny in Nasonia due to haplodiploid sex determination. The cytological events associated with PGL induced by the PSR chromosome and by Wolbachia were compared by fluorescent light microscopy using the fluorochrome Hoescht 33258. Cytological examination of eggs fertilized with PSR-bearing sperm revealed that a dense paternal chromatin mass forms prior to the first metaphase. Quantification of chromatin by epifluorescence indicates that this mass does undergo replication along with the maternal chromatin prior to the first mitotic division but does not replicate during later mitotic cycles. Contrary to previous reports using other staining methods, the paternal chromatin mass remains condensed during interphase and persists over subsequent mitotic cycles, at least until formation of the syncytial blastoderm and cellularization, at which time it remains near the center of the egg with the yolk nuclei. Wolbachia-induced PGL shows several marked differences. Most notable is that the paternal chromatin mass is more diffuse and tends to be fragmented during the first mitotic division, with portions becoming associated with the daughter nuclei. Nuclei containing portions of the paternal chromatin mass appear to be delayed in subsequent mitotic divisions relative to nuclei free of paternal chromatin. Crosses combining incompatibility with PSR were cytologically similar to Wolbachia-induced PGL, although shearing of the paternal chromatin mass was reduced. Wolbachia may, therefore, block an earlier stage of paternal chromatin processing in the fertilized eggs than does PSR. © 1995 Wiley-Liss, Inc.     

Reduced competitive ability due to Wolbachia infection in the parasitoid wasp Trichogramma kaykai

Several hymenopteran parasitoids are infected with parthenogenesis‐inducing (PI) Wolbachia. Infected wasps produce daughters instead of sons from unfertilized eggs. Thus far, little is known about the direct effects of PI Wolbachia on their host's fitness. Here, we report reduced competitive ability due to Wolbachia infection in a minute parasitoid wasp, Trichogramma kaykai Pinto and Stouthamer (Hymenoptera: Trichogrammatidae). Immature survival of infected individuals in a host parasitized by a single infected female, laying a normal clutch of eggs, was lower than those parasitized by a single uninfected individual. When the offspring of infected and uninfected females shared the same host, the infected immatures had significantly lower survival rates than their uninfected counterparts. The survival rate of infected immatures was higher when they competed with other infected immatures from a different infected parent than in competition with uninfected immatures of conspecific wasps. Thus, the host Trichogramma can suffer a substantial reduction in fitness when it is infected with the PI Wolbachia. We discuss why such a reduction is to be expected when populations of infected and uninfected individuals co‐occur, and how the reduced competitive ability of PI Wolbachia influences the spread of the bacteria in the field.     

The paternal sex ratio chromosome in the parasitic wasp Trichogramma kaykai condenses the paternal chromosomes into a dense chromatin mass.

A recently discovered B chromosome in the parasitoid wasp Trichogramma kaykai was found to be transmitted through males only. Shortly after fertilization, this chromosome eliminates the paternal chromosome set leaving the maternal chromosomes and itself intact. Consequently, the sex ratio in these wasps is changed in favour of males by modifying fertilized diploid eggs into male haploid offspring. In this study, we show that in fertilized eggs at the first mitosis the paternal sex ratio (PSR) chromosome condenses the paternal chromosomes into a so-called paternal chromatin mass (PCM). During this process, the PSR chromosome is morphologically unaffected and is incorporated into the nucleus containing the maternal chromosomes. In the first five mitotic divisions, 67% of the PCMs are associated with one of the nuclei in the embryo. Furthermore, in embryos with an unassociated PCM, all nuclei are at the same mitotic stage, whereas 68% of the PCM-associated nuclei are at a different mitotic phase than the other nuclei in the embryo. Our observations reveal an obvious similarity of the mode of action of the PSR chromosome in T. kaykai with that of the PSR-induced paternal genome loss in the unrelated wasp Nasonia vitripennis.     

Effects of deletions on mitotic stability of the Paternal-Sex-Ratio (PSR) chromosome from Nasonia

Paternal-Sex-Ratio (PSR) is a B chromosome that causes all-male offspring in the parasitoid wasp Nasonia vitripennis. It is only transmitted via sperm of carrier males and destroys the other paternal chromosomes during the first mitotic division of the fertilized egg. Because of haplodiploidy, the effect of PSR is to convert diploid (female) eggs into haploid eggs that develop into PSR-bearing males. The PSR chromosome was previously found to contain several families of repetitive DNA, which appear to be present in local blocks. PSR chromosomes with irradiation-induced deletions have decreased rates of transmission and increased variation in transmission. This study investigates whether these differences in transmission of deletion chromosomes are due to mitotic instability. Two deleton chromosomes (E306 and F316) and the wild-type PSR chromosome were examined. A cytogenetic assay of testes revealed that wild-type PSR males contained the chromosome in 98%–100% of their spermatocytes. Similar counts from carriers of two delection chromosomes were lower and varied between individuals from 50%–100%. One F316 male did not contain the chromosome in any of its spermatocytes although the chromosome was present in somatic tissues based on hybridization to PSR-specific repetitive DNA. A molecular analysis of males found the wild-type PSR chromosome to be present in all somatic tissues. Tissue specific differences in the presence of PSR were found in several males from the two deletion lines. The results show that deletions can result in mosaicism due to increased mitotic instability of PSR. Such individuals sometimes partially or completely fail to transmit the chromosome. Patterns of mosaicism of B chromosomes in other organisms are discussed.by P.B. Moens     

The paternal sex ratio chromosome induces chromosome loss independently of Wolbachia in the wasp Nasonia vitripennis

 The paternal sex ratio chromosome (PSR) is a paternally-inherited supernumerary chromosome found in some males of Nasonia vitripennis. PSR induces the loss of N. vitripennis’s paternal autosomes in early fertilized embryos. Previous examinations have not directly addressed the complication of PSR’s co-occurrence with Wolbachia. Wolbachia is the name assigned to a group of cytoplasmic bacteria which induce numerous reproductive alterations in their hosts. In Nasonia, Wolbachia cause cytoplasmic incompatibility (CI) which also results in paternal chromosome loss. Here we address the question of whether PSR’s function (i.e. PSR’s transmission and/or ability to induce chromosome loss) depends upon or interacts with Wolbachia. A strain of PSR males is artificially cleared of Wolbachia. Test crosses and cytological observations of this strain demonstrate that PSR’s transmission and ability to induce chromosome loss is not dependent upon Wolbachia. Comparisons suggest an absence of interactions between PSR and Wolbachia when they co-occur. Fluorescent and confocal microscopy are used to examine and compare early embryos. Observations demonstrate that microtubule interactions with chromatin do not appear to cause the initial loss of the paternal chromosomes. Cytological observations presented here also differ from previous reports of PSR- and Wolbachia-induced chromosome loss. Received: 3 May 1996 / Accepted: 24 June 1996     

A laboratory comparison of four Trichogramma species (Hymenoptera: Trichogrammatidae) as potential biocontrol agents for the carob moth,Ectomyelois ceratoniae (Lepidoptera: Pyralidae)

Four Trichogramma species were recovered in the field from eggs of carob moth, Ectomyelois ceratoniae (Lepidoptera: Pyralidae). The oviposition rates of Trichogramma oleae, Trichogramma cacoeciae, Trichogramma evanescens and Trichogramma bourarachae were compared when presented either E. ceratoniae or Ephestia kuehniella (Lepidoptera: Pyralidae). T. oleae and T. bourarachae did not parasitise either species of Lepidoptera.     

Assessment ofTrichogramma species for biological control of forest lepidopteran defoliators

In a laboratory study, we determined the potential of threeTrichogramma (Hymenoptera: Trichogrammatidae) species,T. brassicae Bezdenko,T. minutum Riley andT. nr.sibiricum Sorokina, for biological control against six species of forest lepidopteran pests, black army cutworm, hemlock looper, eastern spruce budworm, western spruce budworm, white-marked tussock moth, and gypsy moth. Females of each parasitoid species were offered eggs from each of the six host species. Parasitization and the effect of the host species on the emerging progeny were examined and recorded.Trichogramma minutum had the broadest host range and successfully parasitized four host species out of the six offered.Trichogramma nr.sibiricum had the narrowest host range and parasitized only two species of hosts. Of the six host species, black army cutworm was the most preferred by all threeTrichogramma species; white-marked tussock moth and gypsy moth were not parasitized by any parasitoids. There was a positive correlation between the size of female offspring and their corresponding egg complement in all three parasitoid species. The developmental time of parasitoids from egg to adult was influenced by both the parasitoid and host species. Our results suggest thatT. minutum has the greatest potential for biological control against various forest lepidopteran pests and that the black army cutworm may be the best target candidate for further study.     

Biological characteristics of three Trichogramma species on the eggs of diamondback moth (Plutella xylostella L.)

In this study, parasitism rate, development, emergence rate, sex ratio and adult longevity of Trichogramma brassicae, Trichogramma pintoi and Trichogramma embryophagum were investigated on eggs of diamondback moth, Plutella xylostella (Lep. Plutellidae) under controlled conditions. At the beginning of the experiments, 50 eggs of P. xylostella were exposed to parasitism of female wasps of different Trichogramma species. The eggs were replaced daily for three days and exposed eggs were kept in discrete glass vials. According to the obtained results, parasitism rates of T. brassicae, T. pintoi and T. embryophagum were 79.10, 93.90 and 86.10%, respectively. The adult emergence rates showed significant differences among various parasitoid species and were 93.72, 88.16 and 89.93% for T. brassicae, T. pintoi and T. embryophagum, respectively. No significant variations were observed among sex ratio values of the three parasitoid species. Consequently, results of present study suggested that T. pintoi could be more efficient for biocontrol of P. xylostella.     

Host Preferences of Trichogramma pretiosum and the Influence of Prior Ovipositional Experience on the Parasitism of Plutella xylostella and Pseudoplusia includens Eggs

Trichogramma wasps are generalist egg parasitoids used in biological control efforts. In a multi host situation they may preferentially parasitize a non-target host species to the detriment of the control program. Plutella xylostella Linnaeus (Lepidoptera: Plutellidae) is a very serious pest of cabbage, but is only one in a number of species in the ‘cabbageworm’ complex. We investigated the host preferences of Trichogramma pretiosum Riley (Hymenoptera: Chalcidoidea) when offered the eggs of Plutella xylostella and the eggs of Pseudoplusia includens Hübner (Lepidoptera: Noctuidae), another species in the ‘cabbageworm’ complex. Trichogramma pretiosum reared on the eggs of the factitious host Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) parasitized both Plutella xylostella and Pseudoplusia includens eggs under laboratory conditions. For both choice and no-choice experiments, T. pretiosum parasitized significantly more P. xylostella eggs than P. includens eggs. Prior ovipositional experience with one or other of the two host species had no effect on the subsequent parasitism levels of the two host species. The preference for P. xylostella eggs was also not affected by this prior ovipositional experience.     

Aerodynamic separation of parasitised from unparasitised insect host eggs

Aerodynamic sorting of host eggs exposed to Trichogramma egg parasitoids improved the percentage parasitism of retained product. Losses of parasitised eggs declined as percentage parasitism of the unsorted eggs increased. Trichogramma eclosion from parasitised eggs was similar for the unsorted and retained fractions; emergence from the parasitised culls was significantly lower.     

Natural occurrence of Wolbachia-infected and uninfected Trichogramma species in tomato fields in Portugal

Minute egg parasitoids of the genus Trichogramma (Hymenoptera; Trichogrammatidae) are promising candidates for biological control of lepidopteran pests in tomato in Portugal. This certainly applies to native Trichogramma strains that have thelytokous reproduction, i.e., produce only daughters. In Trichogramma wasps, thelytoky is mostly induced by the intracellular bacterium Wolbachia. In this study, we carried out a field survey of native Trichogramma species in four locations in Ribatejo, the main processing tomato region of Portugal, and determined the prevalence of Wolbachia in those species. Five Trichogramma species were found to emerge from lepidopteran eggs collected in the field, namely Trichogramma bourarache, Trichogramma cordubensis, Trichogramma evanescens, Trichogramma pintoi, and Trichogramma turkestanica. T. evanescens and T. pintoi were by far the dominating species representing, respectively, 64.9 and 26.4% of the trichogrammatids collected. Total natural parasitism rates of the collected lepidopteran eggs by Trichogramma wasps ranged from 28.2 to 64.6%. Three Trichogramma species were found to be infected with Wolbachia, namely T. cordubensis, T. evanescens, and T. turkestanica. All the wasp broods belonging to T. cordubensis were infected, whereas low infection rates were found in T. evanescens (0.9% of the broods) and T. turkestanica (4.5% of the broods). The latter represents the first record of a Wolbachia infection in T. turkestanica. Sequencing of the Wolbachia surface protein, wsp, revealed this Wolbachia infection to be related to other Wolbachia infections in Trichogramma wasps. As Wolbachia-infected thelytokous strains exist for T. evanescens, the most abundant Trichogramma species naturally occurring in the tomato fields of the Ribatejo region, this species offers interesting and powerful options for biological control of lepidopteran pests in processing tomato in this region.     

Egg laying and development of Neotropical trichogrammatid species in artificial eggs

The goal of this work was to assess the suitability of two artificial diets for egg laying and development of Trichogramma atopovirilia Oatman & Platner, Trichogrammatoidea annulata De Santis, and Trichogramma bruni Nagaraja (all Hymenoptera: Trichogrammatidae). Additionally, the quality of wasps reared in vitro was compared with those reared in vivo. The ‘standard’ diet consisted of Diatraea saccharalis (Fabricius) (Lepidoptera: Crambidae) pupal holotissues (65%), chicken egg yolk (18%), fetal bovine serum (8.5%), lactalbumin hydrolysate (8.5%), and anticontaminants (0.3%). The ‘modified’ diet differed from the standard one only in the D. saccharalis pupal holotissues, that were replaced with Heliothis virescens (Fabricius) (Lepidoptera: Noctuidae) ones. Females of the three trichogrammatid species laid eggs in artificial eggs containing any artificial medium, but the modified one received more eggs. Although the standard diet was accepted for oviposition by the three wasp species, no development occurred. On the modified diet, only T. atopovirilia was able to develop to adult emergence. Adult F1 were of a quality that was similar to insects reared in vivo.     

Importance of host oviposition pattern and plant size for the selection of Trichogramma strains to control the diamondback moth

The aim of this work was to select a candidate strain of Trichogrammatidae (Hymenoptera: Chalcidoidea) to control the diamondback moth, Plutella xylostella L. (Lepidoptera: Plutellidae), an important pest of cabbage in Europe. The parasitic efficiency of Trichogramma chilonis Ishii from Japan and Réunion Island, Trichogramma evanescens Westwood from Egypt, Trichogramma ostriniae Pang & Chen from Japan, and Trichogramma semblidis (Aurivillius) from France was studied with sentinel eggs in greenhouse‐grown cauliflower. The percentage of parasitized eggs was measured 1 m from the release point and 3 days after release. Two factors were studied: host‐egg density (three eggs per stem vs. 50 eggs per stem) and plant size (50–100 cm high plants vs. 100–150 cm high plants). Trichogramma evanescens from Egypt achieved the best parasitism. Parasitism efficiency of T. ostriniae from Japan and T. chilonis from Réunion Island was not influenced by egg density. In contrast, parasitism efficiencies of the three other strains were higher at the lower egg density than at the higher egg density. Parasitism efficiency was lower when the plants were bigger, with the exception of T. chilonis from Réunion Island, for which the level of parasitism was not affected by plant size. Thus, T. chilonis from Réunion Island appeared to be the most suitable Trichogramma strain for use in controlling P. xylostella, regardless of the growth stage of the crop. However, T. ostriniae from Japan may also be more suitable for use on cauliflower crops at the early stage of crop development. If potential problems related to the introduction of exotic species are taken into consideration, it is possible to consider the native T. evanescens.     

Walking patterns of Trichogramma chilonis and Trichogrammatoidea bactrae upon vegetable leaf surfaces

Trichogramma chilonis Ishii (Hymenoptera: Trichogrammatidae) and Trichogrammatoidea bactrae Nagaraja (Hymenoptera: Trichogrammatidae) are egg parasitoids of the diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae). We observed and recorded the walking patterns of T. chilonis and T. bactrae females on leaves of Raphanus sativus L., Brassica juncea (L.) Czern. et Coss., Brassica campestris L. ssp. chinensis var. utilis Tsen et Lee, and Brassica oleracea L. Our analysis indicated that Trichogramma females spent more time and moved more slowly on R. sativus leaf surface, compared with the other vegetable leaf surfaces. In addition, Trichogramma females were more likely to walk in straight line on B. oleracea leaf surfaces than R. sativus, B. juncea, and B. campestris leaf surface. Compared to T. bactrae females, T. chilonis females spent significantly less time on the leaf surface, and the walking path of T. chilonis was less affected by leaf surface characters (e.g., trichomes, wrinkle, and waxes). During the period of residence on the leaf surface, parasitoid females spent more than 87.8% of their time moving. This study demonstrates that vegetable leaf surface can influence Trichogramma's walking pattern while they are foraging for hosts.     

In vitro cultivation of parthenogenesis-inducing Wolbachia in an Aedes albopictus cell line

Parthenogenesis‐inducing (PI) Wolbachia, infecting the parasitoid wasp Trichogramma kaykai Pinto and Stouthamer (Hymenoptera: Trichogrammatidae), were successfully maintained and cultivated in a mosquito, Aedes albopictus (Skuse) (Diptera: Culicidae), cell line, NIAS‐AeAl‐2. A parthenogenesis‐inducing (PI) Wolbachia strains with wasp ovaries were introduced into flasks with cultures of the cell line. The PI Wolbachia proliferated in NIAS‐AeAl‐2 as confirmed using a quantitative polymerase chain reaction method. Our final goal is to induce parthenogenesis in other hymenopteran species of commercial interest through the artificial transfer of PI Wolbachia. As a step towards this goal, we microinjected pupae of Trichogramma with PI Wolbachia culture in NIAS‐AeAl‐2 cells and PI Wolbachia was detected from recipient by PCR with Wolbachia specific primers (wsp gene).     

Molecular characterization of repetitive DNA sequences from a B chromosome

In the parasitic waspNasonia vitripennis, certain males carry a B chromosome, called PSR (paternal sex ratio), which causes the compaction and subsequent loss of the paternal chromosomes in fertilized eggs. BecauseNasonia are haplo-diploid, this leads to the production of all-male broods. Three families (PSR2, PSR18, PSR22) of related, tandemly repetitive DNAs were shown to be present solely on the PSR chromosome. These three families shared two conserved, palindromic ANA sequences, which may play a role in either PSR function or amplification of the tandem arrays. The tandem repeat family NV79 was determined to be present on the PSR chromosome as well as on at least one of the A chromosomes. This shared repeat as well as two repeat families (NV85, NV126) that were localized on the A chromosomes were detected in two sibling species ofN. vitripennis. NV79 and NV126 were also found in the more distantly related species,Trichomalopsis dubius.by H.F. Willard