Rate Of Development Research Articles

Floral hemp (Cannabis sativa L.) responses to nitrogen fertilization under field conditions in the high desert.

While most studies on floral hemp (Cannabis sativa L.) concur that additions of nitrogen (N) increase plant growth, the performance of floral hemp is heavily influenced by environmental conditions, management and cultivar selection. In regions with a short growing season, the availability of soil N may determine plant developmental rates, final inflorescence biomass and cannabinoid concentrations, but no studies have addressed this for field-grown hemp under high-desert conditions. This field study evaluated the effect of no supplemental N and N fertilization at 90 kg ha-1 on three hemp cultivars (Berry Blossom, Red Bordeaux, and Tahoe Cinco) in Northern Nevada. N increased plant height, canopy cover, stem diameter and shoot biomass, but other physiological parameters were dependent on cultivar. For instance, inflorescence biomass and inflorescence-to-shoot ratio in Red Bordeaux was not affected by N fertilization. Similarly, cannabinoid concentrations were affected by timing of harvest and cultivar but not by N treatment. We evaluated the use of a SPAD meter for ease of determining leaf N deficiency, and correlations with leaf chlorophyll content showed that the SPAD meter was a reliable tool in two cultivars but not in Tahoe Cinco. N treatment increased overall CBD yield, which was driven by increases in inflorescence biomass. Tahoe Cinco was the best CBD yielding cultivar, as it maintained a high inflorescence-to-shoot ratio regardless of N treatment. Our study suggests that even though hemp may have a positive response to soil N management, adjustments based on genotype by environment interaction should be aimed at maximizing cannabinoid yield either by increasing biomass and/or CBD concentrations as long as THC levels are within the permissible <0.3% for U.S. industrial hemp cultivation.

Seed priming with rutin enhances tomato resistance against the whitefly Bemisia tabaci

Flavonoids are ubiquitously distributed in plants, showing pleiotropic effects in defense against abiotic and biotic stresses. Although it has been shown that seed priming with flavonoids can enhance plant resistance to abiotic stress, little is known about its potential to enhance plant tolerance to biotic stresses, especially for herbivorous insects. Here, we investigated whether treatment of tomato (Solanum lycopersicum) seeds with rutin improves plant resistance against the whitefly (Bemisia tabaci). Specifically, we measured the effect of rutin seed treatment on tomato seedling vigour, plant growth, feeding behavior and performance of B. tabaci on plants grown from control and rutin-treated seeds, and plant defense responses to B. tabaci attack. We found that seed treatment with different concentrations of rutin (viz 1, 2, 5, 10, and 20 mM) had minimal impact on shoot growth. Furthermore, seed treatment of rutin reduced the developmental rate of nymphs, the fecundity and feeding efficiency of adult females on plants grown from these seeds. The enhanced resistance of tomato against B. tabaci is closely associated with increased flavonoids accumulation, callose deposition and the expression of jasmonic acid (JA)-dependent defense genes. Additionally, callose deposition and expression of JA-dependent genes in tomato plants grown from rutin-treated seeds significantly increased upon B. tabaci infestation. These results suggest that seed treatment with rutin primes tomato resistance against B. tabaci, and are not accompanied by reductions in shoot growth. Defense priming by seed treatments may therefore be suitable for commercial exploitation.

Emamectin benzoate exposure impaired porcine oocyte maturation

Emamectin benzoate (EB) is a widely used insecticide that can damage the central nervous and immune systems. EB exposure significantly reduced the number of eggs laid, hatching rate, and developmental rate of lower organisms such as nematodes. However, effects of EB exposure on the maturation of higher animals such as porcine oocytes remains unknown. Here we reported that EB exposure severely impaired porcine oocyte maturation. EB exposure with 200 μM prevented cumulus expansion and reduced the rates of first polar body (pb1) extrusion, cleavage and blastocyst after parthenogenetic activation. Moreover, EB exposure disrupted spindle organization, chromosome alignment, and polymerization of microfilaments, but also apparently decreased the levels of acetylated α-tubulin (Ac-Tub) in oocytes. In addition, EB exposure perturbed mitochondria distribution and increased levels of reactive oxygen species (ROS), but did not affect the distribution of cortical granules (CGs) in oocytes. Excessive ROS caused DNA damage accumulation and induced early apoptosis of oocytes. EB exposure led to the abnormal expression of cumulus expansion and apoptosis-associated genes. Altogether, these results demonstrate that EB exposure impaired nuclear and cytoplasmic maturation of porcine oocytes probably through oxidative stress and early apoptosis.

Population performance and detoxifying and protective enzyme activities of four thrips species feeding on flowers of Magnolia grandiflora (Ranunculales: Magnolia).

Different thrips species can co-occur on the same flowers with different dominance degrees. To accurately evaluate the population performance on different thrips species on Magnolia grandiflora flowers, we investigated the diversity of thrips species and their population dynamics both in the field and laboratory. In addition, the activities of detoxifying and protective enzymes in thrips were also measured. Field investigations revealed that four thrips species (Thrips hawaiiensis, Thrips flavidulus, Frankliniella occidentalis, and Thrips coloratus) coexisted on M. grandiflora flowers. They were ranked, from highest population density to lowest, as follows: T. hawaiiensis > T. flavidulus > F. occidentalis > T. coloratus. In laboratory investigations, the species were ranked, from fastest developmental rates to slowest, as follows: F. occidentalis > T. hawaiiensis > T. flavidulus > T. coloratus; and from largest population size to smallest, as follows: T. hawaiiensis > F. occidentalis > T. flavidulus > T. coloratus. Biochemistry assays showed that the four species differed in their activities of detoxifying enzymes (carboxylesterase, glutathione-S-transferase, and cytochrome P450) and protective enzymes (superoxide dismutase, peroxidase) in both laboratory and field strains. Differences in population performance among these four thrips on M. grandiflora may be related to their activity levels of physiological enzymes. The variations in thrips population performance between the field and the laboratory could be due to differences in environmental conditions. T. hawaiiensis showed a strong host preference for M. grandiflora, and thus it has the potential to be a dangerous pest in horticultural plants. © 2023 Society of Chemical Industry.

Life in plastic, it's not fantastic: Sublethal effects of polyethylene microplastics ingestion throughout amphibian metamorphosis

Microplastics (MP) are an abundant, long-lasting, and widespread type of environmental pollution that is of increasing concern as it might pose a serious threat to ecosystems and species. However, these threats are still largely unknown for amphibians. Here, we used the African clawed frog (Xenopus laevis) as a model species to investigate whether polyethylene MP ingestion affects amphibian growth and development and leads to metabolic changes across two consecutive life stages (larvae and juveniles). Furthermore, we examined whether MP effects were more pronounced at higher rearing temperatures. Larval growth, development, and body condition were recorded, and standard metabolic rate (SMR) and levels of stress hormone (corticosterone, CORT) were measured. We determined variation in size, morphology, and hepatosomatic index in juveniles to identify any potential consequences of MP ingestion across metamorphosis. In both life stages, MP accumulation in the body was assessed. MP ingestion was found to result in sublethal effects on larval growth, development, and metabolism, to lead to allometric carry-over effects on juvenile morphology, and to accumulate in the specimens at both life stages. In larvae, SMR and developmental rate increased in response to MP ingestion; there additionally was a significant interaction of MP ingestion and temperature on development. CORT levels were higher in larvae that ingested MP, except at higher temperature. In juveniles, body was wider, and extremities were longer in animals exposed to MP during the larval stage; a high rearing temperature in combination with MP ingestion counteracted this effect. Our results provide first insights into the effects of MP on amphibians throughout metamorphosis and demonstrate that juvenile amphibians may act as a pathway for MP from freshwater to terrestrial environments. To allow for generalizations across amphibian species, future experiments need to consider the field prevalence and abundance of different MP in amphibians at various life stages.

Automated staging of zebrafish embryos using machine learning.

The zebrafish ( Danio rerio), is an important biomedical model organism used in many disciplines, including development, disease modeling and toxicology, to better understand vertebrate biology. The phenomenon of developmental delay in zebrafish embryos has been widely reported as part of a mutant or treatment-induced phenotype, and accurate characterization of such delays is imperative. Despite this, the only way at present to identify and quantify these delays is through manual observation, which is both time-consuming and subjective. Machine learning approaches in biology are rapidly becoming part of the toolkit used by researchers to address complex questions. In this work, we introduce a machine learning-based classifier that has been trained to detect temporal developmental differences across groups of zebrafish embryos. Our classifier is capable of rapidly analyzing thousands of images, allowing comparisons of developmental temporal rates to be assessed across and between experimental groups of embryos. Finally, as our classifier uses images obtained from a standard live-imaging widefield microscope and camera set-up, we envisage it will be readily accessible to the zebrafish community, and prove to be a valuable resource.

Expert consensus on the morphological evaluation of human cleavage-stage embryos and blastocysts.

Expert consensus on the morphological evaluation of human cleavage-stage embryos and blastocysts.

Novel Time-Lapse Parameters Correlate with Embryo Ploidy and Suggest an Improvement in Non-Invasive Embryo Selection.

Selecting the best embryo for transfer is key to success in assisted reproduction. The use of algorithms or artificial intelligence can already predict blastulation or implantation with good results. However, ploidy predictions still rely on invasive techniques. Embryologists are still essential, and improving their evaluation tools can enhance clinical outcomes. This study analyzed 374 blastocysts from preimplantation genetic testing cycles. Embryos were cultured in time-lapse incubators and tested for aneuploidies; images were then studied for morphokinetic parameters. We present a new parameter, "st2, start of t2", detected at the beginning of the first cell cleavage, as strongly implicated in ploidy status. We describe specific cytoplasmic movement patterns associated with ploidy status. Aneuploid embryos also present slower developmental rates (t3, t5, tSB, tB, cc3, and t5-t2). Our analysis demonstrates a positive correlation among them for euploid embryos, while aneuploids present non-sequential behaviors. A logistic regression study confirmed the implications of the described parameters, showing a ROC value of 0.69 for ploidy prediction (95% confidence interval (CI), 0.62 to 0.76). Our results show that optimizing the relevant indicators to select the most suitable blastocyst, such as by including st2, could reduce the time until the pregnancy of a euploid baby while avoiding invasive and expensive methods.

Sex Reversal Induced by Dietary Supplementation with 17α-Methyltestosterone during the Critical Period of Sex Differentiation in Oriental River Prawn (Macrobrachium nipponense).

The steroid 17α-methyltestosterone (MT) inhibits ovarian function and is often used to induce sex reversal artificially in vertebrates. In the present study, different concentrations of MT were added as dietary supplementation, and the effects on sex ratio, growth, and gonadal development were examined. After 40 days, the sex ratio (male:female) in each group increased at different degrees with 50 (1.36:1), 100 (1.57:1), and 200 (2.61:1) mg/kg MT, and neo-males with testis-ovary coexistence were observed in the 200 mg/kg MT group. Furthermore, 50 and 100 mg/kg MT could induce female reversion in neo-males. Histologically, the development of the testes in experimental groups was slower, but the ovaries of the experimental and control groups had similar developmental rates. The expression levels of DMRT11E, Foxl2, and SoxE1 in males at 200 mg/kg MT were 8.65-, 3.75-, and 3.45-fold greater than those of the control group. In crustaceans, sex reversal through vertebrate sex hormones can be observed. Neo-males (sex-reversed female prawns) were maintained by exogenous androgen, and over-reliance led to slow testis growth, small body size, and low growth rate, but sperm was still produced. In female prawns, MT inhibited ovary development and promoted growth.

Detrimental effects of vitrification on integrin genes (α9 and β1) and in vitro fertilization in mouse oocytes.

Integrins are known as key molecules that importantly involve in fertilization. This study aimed to evaluate effects of vitrification on fertilization rate and expression of integrin genes, α9 and β1, on mice oocytes in GV and MІІ stages. From the ovarian tissue and fallopian tube of NMRI mice, germinal vesicle (GV, n = 200) and metaphase II (MII, n = 200) oocytes were obtained. Then, oocytes were distributed into 4 groups including non-vitrified GV, non-vitrified MII, vitrified GV, and vitrified MII. Cryotop method was used for vitrification and oocytes (for 4 weeks) were kept in liquid nitrogen. After that, by using an inverted microscope, the rate of survived oocytes was assessed. Also, in vitro fertilization (IVF) for oocytes, obtained from in vitro maturated MII and mice ovaries (ovulated MII), was done to assess embryos at differenced stages (2-cells, morula, and hatched). Finally, RT-qPCR was performed to investigate the mRNA expression of integrin genes (α9 and β1). After vitrification, the rate of survived oocytes, 68.65%for GV and 65.07% % for MII, did not show a remarkable difference related to non-vitrified groups, while the fertilization rate in vitrified groups remarkably decrease compared to non-vitrified groups (p < 0.05). Also, the expression of α9 and β1 genes was significantly altered in vitrified groups when compared to non-vitrified groups (p < 0.05). There was no significant difference in embryo developmental rates for non-vitrified and vitrified groups. Cryotop method for vitrification caused an alternation in oocyte quality by reducing fertilization rate and integrin gene expression.

Laboratory evaluation of the life history of hibiscus mealybug, Nipaecocus viridis (Hemiptera: Pseudococcidae), on selected citrus and potential non-citrus hosts in Florida.

Nipaecoccus viridis (Newstead) (Hemiptera: Pseudococcidae) is an emerging invasive pest of citrus. There is no information on the life history of N. viridis in relation to citrus cultivars produced in Florida. Here, we quantified the life-history parameters of N. viridis on 6 citrus and 3 non-citrus host plant species under laboratory conditions. We also investigated the effect of mode of reproduction (sexual vs. parthenogenic) on the fecundity of N. viridis. Nipaecoccus viridis was able to develop to maturity and reproduce on all host plant taxa tested. The individual host plants affected the developmental and survival rate, adult longevity, and offspring production of N. viridis. All citrus genotypes commercially grown in Florida were susceptible to N. viridis, with the exception of Citrus aurantium L., which suggests that there is need for management of this invasive pest in Florida citrus groves. Murraya paniculata L., Hibiscus rosa-sinensis L., and Punica granatum L. were also acceptable hosts for N. viridis. Sexually reproducing female N. viridis produced significantly more eggs than those reproducing asexually. Based on our results, propagation of M. paniculata and other citrus relatives should be discouraged around citrus groves in Florida. Given the fitness benefit of sexual reproduction in this pest, pheromone-based tools like mating disruption and mass trapping of adult males could be considered as part of an integrated pest management program with insecticides.

Impact of temperature on Downs herring (Clupea harengus) embryonic stages: First insights from an experimental approach.

Among all human-induced pressures, ocean warming is expected to be one of the major drivers of change in marine ecosystems. Fish species are particularly vulnerable during embryogenesis. Here, the impact of temperature was assessed on embryonic stages of Atlantic herring (Clupea harengus), a species of high socio-economic interest, with a particular focus on the under-studied eastern English Channel winter-spawning component (Downs herring). Key traits linked to growth and development were experimentally evaluated at three temperatures (8°C, 10°C and 14°C), from fertilization to hatching, in standardized controlled conditions. Overall negative impacts of increased temperature were observed on fertilization rate, mean egg diameter at eyed stage, hatching rate and yolk sac volume. A faster developmental rate and a change in development stage frequency of newly hatched larvae were also observed at higher temperature. Potential parental effects were detected for four key traits (i.e. fertilization rate, eyed survival rate, mean egg diameter and hatching rate), despite a limited number of families. For instance, a large variability among families was shown in survival rate at eyed stage (between 0 and 63%). Potential relationships between maternal characteristics and embryo traits were therefore explored. We show that a substantial proportion of variance (between 31 and 70%) could be explained by the female attributes considered. More particularly, age, traits linked to life history (i.e. asymptotic average length and Brody growth rate coefficient), condition and length were important predictors of embryonic key traits. Overall, this study constitutes a stepping-stone to investigate potential consequences of warming on Downs herring recruitment and provides first insights on potential parental effects.

Biological performance and predation rate ofScolothrips longicornisPriesner fed onTetranychus urticaeKoch.

AbstractThe life table and predation rate ofScolothrips longicornisPriesner fed onTetranychus urticaeKoch were examined at 26°C under laboratory conditions. The age‐stage, two‐sex life table was used to analyse the raw data in order to account for the varying developmental rates among individuals and both sexes. The intrinsic rate of increase (r), the finite rate of increase (λ), the net reproductive rate (R0), and the mean generation time (T) were 0.156 day−1, 1.168 day−1and 29.15 offspring, and 21.69 days, respectively.Scolothrips longicornisconsumed 18.40 and 24.47T. urticaeeggs during the first and second larval stages, respectively. The mean consumption rate for females and males during the longevity was 276.26 and 359.79 eggs of Two‐spotted spider mite, respectively. The net consumption rate, which considers survival, was 194.03T. urticaeeggs per individual. The transformation rate fromT. urticaepopulation to predatory thrips offspring was 8.9.

Impact of larvae of the Asian tiger mosquito Aedes albopictus on larvae of the Culex pipiens complex from Germany in laboratory co-breeding studies.

Since the first detection of the Asian tiger mosquito Aedes albopictus (Diptera: Culicidae) eggs in Germany in 2007, several populations of this species have established in Germany. Although colloquially Ae. albopictus is called an 'invasive species', it is not considered 'invasive' and therefore to be controlled according to the European Union (EU) Environmental and Nature Protection Act since evidence of displacement of native species is missing. To test the competitive potential of Ae. albopictus towards mosquito species native to Germany, laboratory experiments were conducted with larvae of this species and indigenous Cx. pipiens complex species/biotypes. First instar larvae of Ae. albopictus and of one of the native taxa were exposed to different temperatures and fed with different food sources. The ratio of individuals developing into adults as well as the time the larvae needed for development were taken as a measure of competitive outcome. In addition, the size of emerging adults was compared between control and experimental groups. Regarding developmental time, no significant differences were found between treatments and controls while significant differences were found regarding developmental rate and average wing size of individuals. Because no evidence of competitive repression of the native species was found, Ae. albopictus cannot be included in the EU list of invasive species.

Gamma Irradiation as a Possible Phytosanitary Treatment against the Greater Wax Moth, Galleria mellonella, Mature Larvae

Larvae of greater wax moth, Galleria mellonella L., can be a devastating pest of honeybee combs in storage. Current control measures may not be adequate or with some drawbacks. Treatment with ionizing radiation could be a possible alternative. Determining the dose, however, requires a detailed study on the radiosensitivity of this stage.&#x0D; In this study, the sensitivity of the greater wax moth mature larvae to ionizing radiation was examined. Larvae were exposed to gamma radiation dosages (50 to 400 Gy) and effects on rate of development, pupation and pupal eclosion (adult emergence) were investigated. In addition, large scale confirmatory tests were conducted to validate treatment efficacy at statistical confidence levels needed for phytosanitary (quarantine and disinfestations) security. Results showed that radiation sensitivity of G. mellonella mature larvae increased, as expected, with increasing radiation dose. The magnitude of the effect, however, depends on the criterion used for measuring effectiveness. Both pupation and pupal eclosion were significantly affected at 50 Gy dose and no emergence was recorded at 300 Gy. Results of large scale confirmatory trials (&gt;30,000 mature larvae) supported the results of the small scale studies. According to probit analysis of dose mortality data for pupation and pupal eclosion, the LD99 for preventing development of mature larvae to pupae and adults is 576 and 318 Gy, respectively. In addition, the developmental rate of mature larvae to adults was negatively affected. This suggests that a dose of 250 Gy may be adequate for disinfesting honeybee products infested with greater wax moth larvae.

Predicting the Occurrence and Risk Damage Caused by the Two-Spotted Spider Mite Tetranychus urticae (Koch) in Dry Beans (Phaseolus vulgaris L.) Combining Rate and Heat Summation Models for Digital Decisions Support

In this work, we use developmental rate models to predict egg laying activity and succession of generations of T. urticae populations under field conditions in the Prespa lakes region in Northern Greece. Moreover, the developmental rate model predictions are related to accumulated heat summations to be compared with actual bean damage and to generate pest-specific degree-day risk thresholds. The oviposition was predicted to start at 57.7 DD, while the first peak in egg laying was estimated to be at 141.8 DD. The second and third peak in egg production were predicted to occur at 321.1 and 470.5 DD, respectively. At the degree-day risk threshold, half development of the first summer generation was estimated at 187 DD and 234 DDm while for the second, it was estimated at 505 DD and 547 DD for 2021 and 2022, respectively. According to the model predictions, no significant differences were observed in the mean generation time (total egg to adult development) of T. urticae between the two observation years (t = 0.01, df = 15, p = 0.992). The total generation time was estimated at 249.3 (±7.7) and 249.2 (±6.7), for 2021 and 2022, respectively. The current models will contribute towards predictions of the seasonal occurrence and oviposition of T. urticae to be used in pest management decision-making. Moreover, the development of population model is a prerequisite for the buildup and implementation of smart plant protection solutions.

LPS Administration during Fertilization Affects Epigenetic Inheritance during Embryonic Development.

Intrauterine inflammation can cause infertility by disrupting reproductive function. The pathogenesis underlying this process may primarily involve endotoxins from lipopolysaccharides (LPS), which are produced by Gram-negative bacteria. However, the long-term effects of endotoxins in mammalian pregnancy following LPS exposure during fertilization have not been clarified. In this study, we performed experiments to analyze the influence of LPS on early embryonic development and fetal development in mice. Mice uteruses were examined for the expression of genes related to the inflammatory response. The expression of Il-1β and Il-6 increased following the administration of 200 and 1000 µg/kg LPS. Exposure to LPS using in vitro fertilization (IVF) significantly decreased the embryonic developmental rate. A concentration of 100 µg/kg LPS significantly increased the placental weight and fetal crown -rump length (CRL), whereas a concentration of 200 µg/kg LPS significantly decreased the placenta weight and fetal weight in vivo. These findings indicate that maternal LPS during fertilization affects fetal development until the late stage of pregnancy. Thus, maternal endotoxins may affect epigenetic inheritance during embryonic development from the early to late stages of pregnancy.

Author Correction: Metabolic regulation of species-specific developmental rates

Author Correction: Metabolic regulation of species-specific developmental rates

Treatments of Porcine Nuclear Recipient Oocytes and Somatic Cell Nuclear Transfer-Generated Embryos with Various Reactive Oxygen Species Scavengers Lead to Improvements of Their Quality Parameters and Developmental Competences by Mitigating Oxidative Stress-Related Impacts.

This study investigated the antioxidant effects of β-cryptoxanthin (BCX), hesperetin (HES), and icariin (ICA), and their effects on in vitro maturation of porcine oocytes and subsequent embryonic development of somatic cell nuclear transfer (SCNT). Treatment with 1 μM BCX (BCX-1) increased the developmental rate of porcine oocytes more than treatment with 100 μM HES (HES-100) or 5 μM ICA (ICA-5). The glutathione level and mRNA expression of antioxidant genes (NFE2L2, SOD1, and SOD2) were more increased in the BCX-1 group than in the HES-100 and ICA-5 groups, while the reactive oxygen species level was more decreased. Moreover, BCX improved the developmental capacity and quality of SCNT embryos. The total cell number, apoptotic cell rate, and development-related gene expression were modulated in the BCX-1 group to enhance embryonic development of SCNT. These results show that the antioxidant effects of BCX enhance in vitro maturation of porcine oocytes and subsequent embryonic development of SCNT.

Differential gene expression and mitonuclear incompatibilities in fast- and slow-developing interpopulation Tigriopus californicus hybrids.

Mitochondrial functions are intimately reliant on proteins and RNAs encoded in both the nuclear and mitochondrial genomes, leading to inter-genomic coevolution within taxa. Hybridization can break apart coevolved mitonuclear genotypes, resulting in decreased mitochondrial performance and reduced fitness. This hybrid breakdown is an important component of outbreeding depression and early-stage reproductive isolation. However, the mechanisms contributing to mitonuclear interactions remain poorly resolved. Here, we scored variation in developmental rate (a proxy for fitness) among reciprocal F2 interpopulation hybrids of the intertidal copepod Tigriopus californicus and used RNA sequencing to assess differences in gene expression between fast- and slow-developing hybrids. In total, differences in expression associated with developmental rate were detected for 2925 genes, whereas only 135 genes were differentially expressed as a result of differences in mitochondrial genotype. Upregulated expression in fast developers was enriched for genes involved in chitin-based cuticle development, oxidation-reduction processes, hydrogen peroxide catabolic processes and mitochondrial respiratory chain complex I. In contrast, upregulation in slow developers was enriched for DNA replication, cell division, DNA damage and DNA repair. Eighty-four nuclear-encoded mitochondrial genes were differentially expressed between fast- and slow-developing copepods, including 12 subunits of the electron transport system (ETS) which all had higher expression in fast developers than in slow developers. Nine of these genes were subunits of ETS complex I. Our results emphasize the major roles that mitonuclear interactions within the ETS, particularly in complex I, play in hybrid breakdown, and resolve strong candidate genes for involvement in mitonuclear interactions.