An Experimental Test of the Relationship between Genetic Variation and Environmental Variation in Tribolium Flour Beetles

The hypothesis that a component of genetic variation for polygenic fitness traits is maintained by environmental heterogeneity was tested using an experimental system involving two species of flour beetles, Tribolium castaneum and T. confusum. Replicated populations of each species from a number of environmental treatments were analyzed for various fitness components following almost 60 generations of natural selection. Environmental differences consisted of flours of cereals commonly invaded by natural populations of these insects.—Tests for adaptation to environments were based on experiments in which populations were reared factorially on each flour, such that population treatment x flour interactions could be detected. Measurements were made of survival, growth rate, larval weight, pupal weight, developmental time, fecundity of individuals at low density and fecundity and cannibalism at high density in both fresh and conditioned media.—Flour differences were found to have significant effects on most traits. Evidence for significant genetic variation and significant genotype x environment interaction was also found. However, no evidence could be found to support the hypothesis that genetic variation was maintained by environmental heterogeneity in food resources. The absence of adaptation to the experimental treatments despite the presence of genetic variation in fitness components suggests that pleiotropy may assume an important role in determining net fitness values of polygenes.     

The Effect of Change in Population Size on DNA Polymorphism

The expected number of segregating sites and the expectation of the average number of nucleotide differences among DNA sequences randomly sampled from a population, which is not in equilibrium, have been developed. The results obtained indicate that, in the case where the population size has changed drastically, the number of segregating sites is influenced by the size of the current population more strongly than is the average number of nucleotide differences, while the average number of nucleotide differences is affected by the size of the original population more severely than is the number of segregating sites. The results also indicate that the average number of nucleotide differences is affected by a population bottleneck more strongly than is the number of segregating sites.     

The Effect of Nonindependent Mate Pairing on the Effective Population Size

The effective population size (N_e) quantifies the effectiveness of genetic drift in finite populations. When generations overlap, theoretical expectations for N_e typically assume that the sampling of offspring genotypes from a given individual is independent among successive breeding events, even though this is not true in many species, including humans. To explore the effects on N_e of nonindependent mate pairing across breeding events, we simulated the genetic drift of mitochondrial DNA, autosomal DNA, and sex chromosome DNA under three mating systems. Nonindependent mate pairing across breeding seasons has no effect when all adults mate pair for life, a small or moderate effect when females reuse stored sperm, and a large effect when there is intense male–male competition for reproduction in a harem social system. If adult females reproduce at a constant rate irrespective of the type of mate pairing, the general effect of nonindependent mate pairing is to decrease N_e for paternally inherited components of the genome. These findings have significant implications for the relative N_e values of different genomic regions, and hence for the expected levels of DNA sequence diversity in these regions.THE effective population size (N_e) is a fundamental parameter of population genetics, which quantifies the effect of genetic drift, the stochastic change in allele frequencies over time in a population of finite size (Wright 1931). The magnitude of N_e affects both the level of genetic variability within a population and the efficiency with which populations retain mildly beneficial mutations and purge mildly deleterious ones. This influences a myriad of genetic phenomena, such as the level of DNA sequence polymorphism, the rate of substitution of nonsynonymous and functional noncoding nucleotide positions, the abundance of transposable elements, levels of variation, and the rate of evolution of gene expression, the persistence of duplicate genes, and genome size and organization (Lynch 2007; Charlesworth 2009). There are a variety of definitions of N_e; here we use the definition in terms of the mean coalescence time of a pair of neutral alleles, which is given by 2N_e (Charlesworth 2009). This definition has the useful feature that the expected pairwise nucleotide site diversity under the widely used infinite sites model is equal to 4N_eμ, where μ is the neutral mutation rate (Kimura 1971).As a result of differences in their ploidy level and mode of inheritance, autosomal DNA (aDNA), the X chromosome (xDNA), the Y chromosome (yDNA), and maternally transmitted organelle DNA such as mitochondrial DNA (mtDNA) generally have a different N_e values. Under certain conditions, such as constant population size, discrete generations, a Poisson distribution of reproductive success, and a sex ratio equal to one, the relative N_e values of these genomic regions (N_e-a, N_e-x, N_e-mt, and N_e-y) are expected to be 4:3:1:1 (Charlesworth 2009). This is because aDNA is biparentally inherited and diploid; xDNA is biparentally inherited, diploid in females, and haploid in males (with female heterogamety, the reverse applies to the Z chromosome), and yDNA (the W chromosome, with female heterogamety) and mtDNA are both effectively uniparentally inherited and haploid in most species.However, several characteristics of natural populations, such as unequal numbers of males and females and nonrandom variation in reproductive success, can affect the value of N_e, even for populations with discrete generations (reviewed in Caballero 1994; Hedrick 2007; Charlesworth 2009). In addition, natural selection at sites linked to neutral markers also has the potential to increase N_e (under balancing selection) (Charlesworth 2006) or to decrease N_e (with background selection or selective sweeps) (Hudson and Kaplan 1988; Charlesworth et al. 1993). Because the nature of natural selection varies in different genomic regions, especially in relation to the rate of recombination, N_e may also vary among unlinked regions with the same ploidy and mode of inheritance, for example, different portions of an autosomal chromosome (Gossmann et al. 2011). In natural populations, these factors can skew the relative N_e values away from the 4:3:1:1 expectation. Even when the effects of natural selection and “nonideal” demography are ignored, the 4:3:1:1 relation still has a large variance when applied to individual loci (Hudson and Turelli 2003).When generations overlap, an additional source of possible deviations from these idealized relations arises from variation among individuals in survival and reproductive success among breeding seasons (Felsenstein 1971; Hill 1972, 1979; Johnson 1977) and from sex differences in demographic parameters and stochastic changes in population size (Engen et al. 2007). In contrast, the effect of a high variance in reproductive success caused by male–male competition is lessened when generations overlap for many breeding seasons (Nunney 1993; Charlesworth 2001). Conversely, overlapping generations with nonindependent mate pairing across breeding seasons could increase the variance in reproductive success. For example, in humans, paternity is correlated with paternal confidence in paternity (Anderson 2006), and married individuals tend to repeatedly produce offspring with each other more frequently than expected by chance. Nonindependent mate pairing among breeding seasons occurs in many other species as well—for example, long-term pair bonding in prairie voles (DeVries et al. 1995), harems in gorillas (Gatti et al. 2004), and sperm storage in fruit flies (Neubaum and Wolfner 1999).Current theoretical models that allow calculation of the effective population size with overlapping generations and age structure make several simplifying assumptions, notably constant sizes of each age class, sufficiently large numbers of individuals in each age class that second-order terms in their reciprocals can be neglected, and independent sampling of offspring genotypes from the same individual reproducing at different times (Hill 1972; Nunney 1991, 1993; Caballero 1994; Charlesworth 1994, 2001). The latter assumption in particular makes it difficult to provide accurate expressions for species such as humans and Drosophila, which reproduce nonindependently because of long-term pair bonds and sperm storage, respectively (Charlesworth 2001).The goal of this study is therefore to explore the consequences of nonindependence of reproductive events across time in different social systems and with different age structures, using simulations of genetic drift in two types of age-structured populations, under different scenarios of independent and nonindependent mate pairing among breeding events. We have explored how these scenarios affect the relative values of N_e-a, N_e-x, N_e-mt, and N_e-y using the infinite alleles model of mutation (Kimura and Crow 1964), with particular emphasis on comparisons of similar mating systems that differ in the extent of nonindependence among breeding events.     

The Effect of an Upper Limit to Population Size on Persistence Time

For a population with density-independent vital rates in a randomly varying environment, previous authors have calculated the probability that population size will first drop to some specified (arbitrary) low level at a given time (the first passage time distribution (FPTD), which may be interpreted as a distribution of extinction times). In this paper, we study the FPTD For a stochastic model of density-independent population growth which includes a hard upper limit to population size. We discuss the conditions under which this distribution may be approximated by the FPTD of a Wiener process with a reflecting boundary condition, for which an exact calculation is presented in an appendix. We compare the FPTD of the new model with its counterpart in the model without an upper limit. The most important effects of introducing the upper limit are: (a) ultimate extinction becomes certain; (b) if the long run growth rate in the absence of the upper boundary was small but positive, extinction within ecologically significant times is likely; (c) for larger values of the long run growth rate, persistence over ecologically significant times is almost certain. We discuss the implications of result (b) for conservation. Result (c) establishes that "density-vague" regulation can produce persistent, but bounded, populations.     

Critical Mutation Rate Has an Exponential Dependence on Population Size in Haploid and Diploid Populations

Understanding the effect of population size on the key parameters of evolution is particularly important for populations nearing extinction. There are evolutionary pressures to evolve sequences that are both fit and robust. At high mutation rates, individuals with greater mutational robustness can outcompete those with higher fitness. This is survival-of-the-flattest, and has been observed in digital organisms, theoretically, in simulated RNA evolution, and in RNA viruses. We introduce an algorithmic method capable of determining the relationship between population size, the critical mutation rate at which individuals with greater robustness to mutation are favoured over individuals with greater fitness, and the error threshold. Verification for this method is provided against analytical models for the error threshold. We show that the critical mutation rate for increasing haploid population sizes can be approximated by an exponential function, with much lower mutation rates tolerated by small populations. This is in contrast to previous studies which identified that critical mutation rate was independent of population size. The algorithm is extended to diploid populations in a system modelled on the biological process of meiosis. The results confirm that the relationship remains exponential, but show that both the critical mutation rate and error threshold are lower for diploids, rather than higher as might have been expected. Analyzing the transition from critical mutation rate to error threshold provides an improved definition of critical mutation rate. Natural populations with their numbers in decline can be expected to lose genetic material in line with the exponential model, accelerating and potentially irreversibly advancing their decline, and this could potentially affect extinction, recovery and population management strategy. The effect of population size is particularly strong in small populations with 100 individuals or less; the exponential model has significant potential in aiding population management to prevent local (and global) extinction events.     

山地次生林破碎化对喜鹊繁殖功效的影响

于1999－2000年春夏两季,在左家自然保护区及土门岭地区研究了破碎化山地次生林斑块面积对喜鹊（Pica pica）繁殖功效的影响。运用GPS定位系统确定了12块大（＞20hm^2,n=4）、中（10－20hm^2,n=hm^2）对照样地（每块面积20hm^2）。观测了各斑块及对照样地中喜鹊产第1枚卵的时间、窝卵数、平均卵重、出雏量及雏鸟出飞量等生态指标,结果表明,（1）斑块面积对喜鹊的繁殖功效存在着显著的影响;（2）喜鹊在对照样地和大面积斑块中的平均产卵时间早于小斑块中的11.62d;（3）对照样地和大面积斑块内的窝卵数略高于中、小面积块内,但差异不显著;（4）各斑块及对照样地中的平均卵重无显著差异,小面积斑块中的出雏率和雏鸟出飞率均最低（51.7%和42.88%）,大面积斑块中的出雏率最高（72.12%）,对照样地略次之（71.93%）,对照样地中的雏鸟出飞率最高（65.45%）,大面积斑块次之（62.71%）;（5）喜鹊在小面积次生林斑块中繁殖功效较低的主要原因是巢损失率较高。     

Rate of Decrease of Genetic Variability in a Two-Dimensional Continuous Population of Finite Size

The rate of decay of genetic variability was investigated for two-dimensional continuous populations of finite size. The exact value of the rate involves a rather complicated expression (formula (4-1)). However, numerical examples indicate that in a population habitat size LxL and density D, the rate is approximately equal to (see PDF) where sigma(2) is the variance of dispersion distance assuming isotropical migration. The value given in (2) is equal to that of a panmictic population of size DL(2). It is remarkable that whether the rate assumes the value given by (1) or by (2) depends only on Dsigma(2) (a local property), which is independent of the habitat size. Since, in a one-dimensional population, this depends on both Dsigma(2) and the habitat size, there is an essential difference between the two types of population structure.-The function giving the probability of two homologous genes separated by a given distance being different alleles was also obtained, (formula (5-1)).     

Directional versus Stabilizing Selection for Developmental Time in Natural and Laboratory Populations of Flour Beetles

Artificial selection for fast development is successful in long-established laboratory populations of Tribolium, but not in strains recently derived from natural populations. It is shown that selection against fast development in dense, synchronized cultures operates through cannibalism of early pupae by larvae. Since standard husbandry procedures for laboratory strains involve the periodic creation of dense, synchronized cultures, it is suggested that these populations are subjected to stabilizing selection for intermediate developmental time. Natural populations, on the other hand, are probably subjected to directional selection for rapid development.     

Effect of Group Size on Feeding Rate when Patches are Exhaustible

One benefit of group foraging is that individual foragers can join the food discoveries of companions and thus increase encounter rate with food patches. When food patches are exhaustible, however, individual shares of each patch will decrease with group size negating the effect of increased encounter rate. Mean feeding rate may actually decrease with group size as a result of aggression or time wasted joining already depleted patches, or when searching to join the food discoveries of others, which is referred to as scrounging, precludes finding food. I examined the relationship between mean feeding rate and group size in captive flocks of zebra finches (Taenopygia guttata) foraging for small clumps of seeds. Finches in groups of two or four fared better than solitary birds in terms of mean feeding rate despite the fact that birds in groups scrounged a large proportion of their food. Solitary birds initiated feeding activity after a longer delay, which led to their lower success. Early departures by food finders from food patches joined by others may have lessened the impact of scrounging on mean feeding rate. As a result of benefits from the presence of companions, group foraging in zebra finches appears a viable alternative to foraging alone despite the cost of sharing resources.     

炔雌醚对长爪沙鼠种群数量和性比的影响

炔雌醚是一种激素类新型鼠类不育剂,已有研究表明,炔雌醚对某些鼠类种群繁殖具有良好的控制成效。为检验新型不育剂对长爪沙鼠种群的控制效果,2006年5月在内蒙古锡林郭勒盟锡林浩特市农牧交错带开展了炔雌醚对野外长爪沙鼠种群的控制实验,本次实验设投药区和对照区2个组别,5—8月开展逐月的夹线调查以及四分之一圆洞口计数法,对样地内长爪沙鼠进行系统的追踪调查,分析炔雌醚对长爪沙鼠种群数量以及种群性比的作用。结果显示:投药1个月之后,投药区雌鼠比例(16.3%)不及对照组雌鼠比例(51.9%)的三分之一,投药区的雌鼠比例(16.3%)仅为雄鼠比例(83.7%)的五分之一;与对照区相比,投药区长爪沙鼠的密度下降达60%以上。这表明:炔雌醚对野外长爪沙鼠种群的不育控制具有良好效果,且炔雌醚对长爪沙鼠的种群性比影响很大,过量投放炔雌醚可造成雌鼠死亡,单独使用炔雌醚可降低野外不育剂的药物成本。     

Effect of Dominance,Reproductive State,and Group Size on Body Mass in Macaca radiata

Body mass can impact reproductive performance in males and females. In nonhuman primates the relationship is often mediated by dominance. We measured body mass monthly in a provisioned group of bonnet macaques (Macaca radiata) living at a Hindu temple. We also measured body mass on 3 occasions in a wild population of bonnet macaques. In the temple group, females that reproduced lost body mass, while females that did not reproduce gained body mass. Mass loss among females occurred primarily while they were nursing. Adult males from the temple group lost mass during the mating season and gained it during the non-mating season. Subadult males experienced less seasonal variation in body mass. Body mass and changes in mass were not related to dominance rank in either the temple or the forest group. Furthermore, maternal dominance rank did not affect infant mass. Females from the smallest forest group weighed significantly less than females from the two larger forest groups, which suggests intergroup competition in the population. Body mass was not related to dominance rank in a straightforward manner but may indirectly affect reproductive performance. The pattern of body mass change suggests that the period of lactation is critical for females and endurance rivalry is an important form of competition among males.     

Some Statistical Improvements for Estimating Population Size and Mutation Rate from Segregating Sites in DNA Sequences

In population genetics, under a neutral Wright-Fisher model, the scaling parameter straight theta=4Nmu represents twice the average number of new mutants per generation. The effective population size is N and mu is the mutation rate per sequence per generation. Watterson proposed a consistent estimator of this parameter based on the number of segregating sites in a sample of nucleotide sequences. We study the distribution of the Watterson estimator. Enlarging the size of the sample, we asymptotically set a Central Limit Theorem for the Watterson estimator. This exhibits asymptotic normality with a slow rate of convergence. We then prove the asymptotic efficiency of this estimator. In the second part, we illustrate the slow rate of convergence found in the Central Limit Theorem. To this end, by studying the confidence intervals, we show that the asymptotic Gaussian distribution is not a good approximation for the Watterson estimator.     

The Functional Transfer of Genes From the Mitochondria to the Nucleus: The Effects of Selection,Mutation, Population Size and Rate of Self-Fertilization

The transfer of mitochondrial genes to the nucleus is a recurrent and consistent feature of eukaryotic genome evolution. Although many theories have been proposed to explain such transfers, little relevant data exist. The observation that clonal and self-fertilizing plants transfer more mitochondrial genes to their nuclei than do outcrossing plants contradicts predictions of major theories based on nuclear recombination and leaves a gap in our conceptual understanding how the observed pattern of gene transfer could arise. Here, with a series of deterministic and stochastic simulations, we show how epistatic selection and relative mutation rates of mitochondrial and nuclear genes influence mitochondrial-to-nuclear gene transfer. Specifically, we show that when there is a benefit to having a mitochondrial gene present in the nucleus, but absent in the mitochondria, self-fertilization dramatically increases both the rate and the probability of gene transfer. However, absent such a benefit, when mitochondrial mutation rates exceed those of the nucleus, self-fertilization decreases the rate and probability of transfer. This latter effect, however, is much weaker than the former. Our results are relevant to understanding the probabilities of fixation when loci in different genomes interact.GENOMIC investigations of obligate intracellular endosymbionts (Moran and Wernegreen 2000; Akman et al. 2002; Tamas et al. 2002; Wernegreen et al. 2002; Klasson and Andersson 2004; Foster et al. 2005) reveal a reduction in genome size and number of protein-coding genes compared to their free-living relatives (Charles et al. 1999; Gil et al. 2002; Wernegreen et al. 2002; Moran 2003; Van Ham et al. 2003; Klasson and Andersson 2004; Khachane et al. 2007). Similarly, mitochondria—ancient obligate intracellular symbionts of eukaryotes—have retained very few protein-coding genes (Boore 1999; Adams et al. 2002) [Reclinomonas americanas is at the extreme of retention of mitochondrial genes (Lang et al. 1997)]. Understanding the process of gene loss in mitochondria and other endosymbionts is a major research focus of mitochondrial and endosymbiont genomics (Moran 2003; Timmis et al. 2004; Khachane et al. 2007; Bock and Timmis 2008).The loss of endosymbiont genes can be complete, in which lost genes are absent from the host–endosymbiont complex, a substitution, in which a nuclear allele functions in place of the lost symbiont gene, or a functional transfer of an endosymbiont gene to the nucleus, followed by its loss (Adams and Palmer 2003). Such “functional transfer” involves the relocation of a mitochondrial gene to the nucleus, its acquisition of a promoter, successful targeting to the mitochondria for proper function, and the eventual loss of the gene from the mitochondrial genome altogether. Although this process is probably quite complex and requires numerous evolutionary modifications (Murcha et al. 2005), there is evidence that some mitochondrial genes are preadapted to functional transfer as they contain signals that target them to the mitochondria before functional transfer to the nucleus (Ueda et al. 2008a). The complex evolution of rps16 is an illuminating case of both functional gene transfer and substitution. In some lineages, the mitochondrial rps16 is functionally expressed in the nucleus but absent from the mitochondria (functional transfer) while in a subset of taxa, the chloroplast copy is also absent and the nuclear gene is also targeted to the chloroplast [substitution (Ueda et al. 2008b)].A number of evolutionary scenarios have been proposed to account for the massive loss of genes from endosymbionts. A subset of models argues that endosymbiont gene loss is a neutral or nearly neutral process. Since endosymbiosis reduces the strength of selection on genes that are unnecessary or redundant in an obligate intracellular environment, these genes may be quickly lost by the neutral fixation of a deletion or other loss-of-function mutations. Moreover, even when selection favors the retention of genes in endosymbionts, such selection may be ineffective because of reduction in effective population size due to recurrent bottlenecking (Rispe and Moran 2000). Additionally, frequent input of functional endosymbiont genes into the nucleus makes symbiont genes redundant, exacerbating gene loss via functional transfer (Berg and Kurland 2000).An alternative class of explanations views the loss of mitochondrial genes (be it complete loss, substitution, or functional transfer) as an adaptive process. The “mitochondrial competition theory” argues that mitochondrial genomes that either do not contain or do not express a given allele have a replicative advantage over other mitochondria, providing a within-host selective advantage to mitochondrial gene loss (Albert et al. 1996; Selosse et al. 2001; Yamauchi 2005). The “benefits of sex” model posits that the genomic diploid nuclear environment (diploid, sexual) is in some way preferable (e.g., as an escape from Muller''s ratchet or Hill–Robertson interference) to a haploid asexual mitochondrial environment (Blanchard and Lynch 2000). The epistatic model (Wade and Goodnight 2006) does not advance a specific or consistent benefit to transfer, but posits that transfer is explicitly a process of coevolution between mitochondrial and nuclear genomes, where fitness is a function of the gene combination rather than of either gene separately.Because few species are currently undergoing mitochondrial to nuclear gene transfer, these alternative hypotheses are difficult to distinguish with direct experimentation. However, the distribution of transferred genes across lineages allows for evaluation of the alternative hypotheses. For example, self-pollination reduces the rate of heteroplasmy and consequently the opportunity for competition among genetically distinct mitochondria. Thus, the mitochondrial competition theory predicts an excess of transfer events in sexual, outcrossing lineages, with high degrees of “paternal leakage.” Similarly, frequent self-fertilization diminishes the benefits of sex, and thus the benefits of sex hypothesis predicts fewer transfers in selfing and clonally reproducing plants than in outcrossing taxa. The epistatic model makes the opposite prediction. Selfing and clonal reproduction maintain cyto-nuclear gene combinations and increase the response to selection on epistatic combinations, potentially encouraging transfer. On the other hand, outcrossing tends to break apart adaptive cyto-nuclear gene combinations, potentially decreasing the amount of adaptive transfer in outcrossing lineages.Plant lineages with high levels of self-fertilization or asexual reproduction transfer more mitochondrial genes to their nuclei than predominantly sexual and outcrossing lineages (Brandvain et al. 2007). This result is consistent with predictions of the epistatic model and is contrary to predictions of the mitochondrial competition or benefits of sex models. More specific predictions allowing further empirical tests require more detailed theoretical investigations of the gene transfer process. Here, we investigate the roles of mutation, selection, and random drift in gene transfer using both deterministic models and stochastic simulations to refine and extend predictions of patterns of functional mitochondrial to nuclear gene transfer.     

Assessing the Effect of Sequencing Depth and Sample Size in Population Genetics Inferences

Next-Generation Sequencing (NGS) technologies have dramatically revolutionised research in many fields of genetics. The ability to sequence many individuals from one or multiple populations at a genomic scale has greatly enhanced population genetics studies and made it a data-driven discipline. Recently, researchers have proposed statistical modelling to address genotyping uncertainty associated with NGS data. However, an ongoing debate is whether it is more beneficial to increase the number of sequenced individuals or the per-sample sequencing depth for estimating genetic variation. Through extensive simulations, I assessed the accuracy of estimating nucleotide diversity, detecting polymorphic sites, and predicting population structure under different experimental scenarios. Results show that the greatest accuracy for estimating population genetics parameters is achieved by employing a large sample size, despite single individuals being sequenced at low depth. Under some circumstances, the minimum sequencing depth for obtaining accurate estimates of allele frequencies and to identify polymorphic sites is , where both alleles are more likely to have been sequenced. On the other hand, inferences of population structure are more accurate at very large sample sizes, even with extremely low sequencing depth. This all points to the conclusion that under various experimental scenarios, in cost-limited population genetics studies, large sample sizes at low sequencing depth are desirable to achieve high accuracy. These findings will help researchers design their experimental set-ups and guide further investigation on the effect of protocol design for genetic research.     

Dissecting Systemic RNA Interference in the Red Flour Beetle Tribolium castaneum: Parameters Affecting the Efficiency of RNAi

The phenomenon of RNAi, in which the introduction of dsRNA into a cell triggers the destruction of the corresponding mRNA resulting in a gene silencing effect, is conserved across a wide array of plant and animal phyla. However, the mechanism by which the dsRNA enters a cell, allowing the RNAi effect to occur throughout a multicellular organism (systemic RNAi), has only been studied extensively in certain plants and the nematode Caenorhabditis elegans. In recent years, RNAi has become a popular reverse genetic technique for gene silencing in many organisms. Although many RNAi techniques in non-traditional model organisms rely on the systemic nature of RNAi, little has been done to analyze the parameters required to obtain a robust systemic RNAi response. The data provided here show that the concentration and length of dsRNA have profound effects on the efficacy of the RNAi response both in regard to initial efficiency and duration of the effect in Tribolium castaneum. In addition, our analyses using a series of short dsRNAs and chimeric dsRNA provide evidence that dsRNA cellular uptake (and not the RNAi response itself) is the major step affected by dsRNA size in Tribolium. We also demonstrate that competitive inhibition of dsRNA can occur when multiple dsRNAs are injected together, influencing the effectiveness of RNAi. These data provide specific information essential to the design and implementation of RNAi based studies, and may provide insight into the molecular basis of the systemic RNAi response in insects.