Phenotypic Similarities and Differences of Genetic Characteristic Boer and PE Goat Yield Crossbreeding 1st and 2nd Generation

This research aimed to analyze the phenotypic similarity and genetic characteristics of 1st and 2nd generation (G1 and G2) goat of Boer and PE goat yield crossbreeding. The number of offsprings of G1 and G2 was46 and 28 goats, respectively. Data analysis of phenotypic characteristics was estimated by calculating the opportunities of the number of the appearance of G1 and G2 offspring. The result showed that the percentages of the G2 of W11 similarity (67.86%) more significant than G1 (26.09%). The W21was the G1 (45.65%) higher than G2 (10.71%), and W31 was G1 (28.26%) higher than G2 (21.43%). It concluded that G2 had phenotypic similarities of the characteristic (color) in total progeny. The G2 goat displays the phenotypic characters of inheritance (color)patterns that had more significant similarity than the results of the G1, as well as similarities in morphological characters that were different from the G1. The characteristics (colors) in the G2 had dark brown head color greater than G1, and body conformation approaches to Boer goat. Other colors like light brown, plain white, black, and straight black on G1 greater than G2.

Life History and Rearing of Anastatus orientalis (Hymenoptera: Eupelmidae), an Egg Parasitoid of the Spotted Lanternfly (Hemiptera: Fulgoridae)

To support efforts to manage and contain spotted lanternfly (SLF), Lycorma delicatula White (Hemiptera: Fulgoridae), research is being conducted to develop classical biological control methods. To date, two potential biocontrol agents from China have been identified: an egg parasitoid, Anastatus orientalis, and a nymphal parasitoid, Dryinus sinicus Olmi (Hymenoptera: Dryinidae). The research detailed here focuses on investigating the biology and rearing of A. orientalis to assess its potential efficacy in a biocontrol program and optimize its rearing. Female wasps lived significantly longer than male wasps (68 and 23 d, respectively) and females produced an average of 94 total progeny that successfully emerged as adults, with most progeny produced between weeks one and four of the females’ lives. The sex ratio of the progeny, with no re-mating, was initially highly female-biased but became progressively more male-biased, likely due to sperm depletion. There was no evidence of additional mortality to SLF eggs from wasp host feeding, but the data were highly variable and the sample size was small. There was high parasitoid emergence when oviposition conditions mimicked mid-September Beijing temperature and photoperiod; however, there was little emergence under 25°C and long-day conditions because most progeny entered a diapause. Storage of parasitized eggs in 5°C chill lowered parasitoid emergence rates. Lastly, there was no evidence that storing field-collected SLF egg masses in 5°C for 10 mo prior to parasitization affected parasitism rates. These findings inform our rearing protocol for A. orientalis and facilitate our testing of this species as a potential biological control agent for SLF.

Construction of a Baculovirus Derivative to Produce Linearized Antheraea pernyi (Lepidoptera: Saturniidae) Multicapsid Nucleopolyhedrovirus Genomic DNA

In the Antheraea pernyi multicapsid nucleopolyhedrovirus (AnpeNPV)-based expression vector system, the frequency of homologous recombination events between wild-type AnpeNPV DNA and the transfer vector is low, resulting in a small amount of recombinant virus. Previous reports have indicated that linearized baculovirus DNA can increase the proportion of recombinant virus relative to the total progeny. To improve the recombination efficiency, we constructed a linearized derivative of AnpeNPV, referred to as AnpeNPVPhEGFP-AvrII, in which egfp flanked by AvrII restriction sites was located at the polyhedrin locus and driven by the polyhedrin promoter. Linear AnpeNPV DNA was obtained by the treatment of AnpeNPVPhEGFP-AvrII genomic DNA with AvrII endonuclease. The infectivity and recombinogenic activity between the linearized and circular viral DNA were evaluated by quantitative real-time polymerase chain reactions. We demonstrated that the linearized AnpeNPV DNA produced only small numbers of infectious budded viruses, accounting for approximately 4.5% of the budded virus production of wild-type AnpeNPV DNA in A. pernyi pupae. However, the linearized AnpeNPV DNA substantially increased recombinant virus production after cotransfection with an appropriate transfer vector; relative abundance of the recombinant virus was approximately 5.5-fold higher than that of the wild-type AnpeNPV DNA in A. pernyi pupae. The linearization of AnpeNPV DNA will facilitate the purification of recombinant viruses using the AnpeNPV-based expression vector system and the construction of an AnpeNPV-based bacmid system.

Characterization of New Tea (Camellia sinensis L.) Hybrid Progenies Based on Morphological Traits

Hybridization is the main method of obtaining genetic variation and breeding of new cultivars. Two parents selected for their desired characteristics were crossed to create genetic variation. One hundred and eighteen putative hybrids were generated by crossing two diverse parents, TRI 2043 which is characterized with high pubescence density, pigmented leaves, tolerant to blister blight disease, and TRI 3055 a non-pigmented, stem canker tolerant cultivar. A reciprocal cross was also made. Total progeny was characterized for five morphological traits; anthocyanin pigmentation in petiole, leaf vestiture, average number of pubescence of leaf, immature leaf colour, and petiole colour. Average number of pubescence in abaxil of the second leaf varied from 5 to 149 with the mean of 62. Parental cultivar TRI 2043 recorded the highest average number of pubescence and TRI 3055 had the lowest. Among the progenies, 93 individuals contained anthocyanin pigmentation in petiole, which was the characteristic feature of TRI 2043. Population was subjected to genetic analysis and the resultant dendrogram clearly categorized the progenies into four clusters. Significant variation was found among the individuals in the progeny. Forty individuals showed close resemblance with the parent TRI 2043 and 21 individuals grouped with cultivar TRI 3055. Rest of the individuals shared both parental morphological characteristics.

Abstract 42: Genetic Ablation of TMEM16F Exhibits Strain-specific Lethality in Mice

Background: Scott Syndrome is a rare bleeding disorder characterized by a defect in platelet phosphatidylserine (PS) exposure. The syndrome has recently been linked to mutations in TMEM16F. Tmem16f -/- mice were recently reported to be viable with a prolonged tail snip bleeding time but no spontaneous bleeding. We now report analysis of an additional gene targeted Tmem16f allele generated in C57BL/6 ES cells. Results: JM8 ES cell were obtained from EUCOMM, and successful Tmem16f gene targeting in intron 1 was confirmed by PCR and sequencing. Genotyping of 120 Tmem16f +/gt (+/gt) intercross progeny identified no surviving Tmem16f gt/gt (gt/gt) mice at weaning (p<0.001). However, +/gt intercrosses generated the expected Mendelian genotype ratios at both E10.5 and E17.5, with gt/gt embryo’s exhibiting no morphological abnormalities on gross or routine histologic examination. Though complete deficiency of TMEM16F is lethal in the C57BL/6J genetic background between E17.5 and birth, an F2 intercross of +/gt mice outcrossed one generation to 129x1SvJ resulted in gt/gt mice surviving to weaning, though at reduced numbers (6/75 total progeny compared to ~19 expected, p <0.002). Progeny testing of surviving gt/gt mice suggest a single autosomal dominant 129x1SvJ-associated genetic modifier. Preliminary genetic analysis of these mice appears to map this locus to the proximal region of chromosome 3. Tail bleeding times for gt/gt were >10min, whereas littermate +/gt and +/+ mice bleeding ceased at 8 ± 1 min and 6 ± 0.8 min, respectively, each significantly different than gt/gt (p<0.05). Notably, platelets from +/gt mice exhibited a trend toward reduced PS exposure, detected with FITC-labelled lactadherin, in response to PAR4 agonist peptide, whereas gt/gt mice had significantly reduced PS exposure (p < 0.05). Conclusion: These data suggest the existence of a viability-determining genetic modifier of TMEM16F in the 129x1SvJ mouse strain. Identification of the responsible gene may uncover novel functions for TMEM16F and the regulation of hemostatic function.

TMEM16F Deficiency Causes a Strain-Dependent Lethality in Mice and Reduced Phosphatidylserine Exposure on Heterozygous As Well As Homozygous Deficient Mouse Platelets

Scott Syndrome is a rare bleeding disorder characterized by a defect in platelet phosphatidylserine (PS) exposure. The syndrome has recently been linked to mutations in TMEM16F, a Ca++-activated ion channel. Tmem16f-/- mice were recently reported to be viable with a prolonged tail snip bleeding time but no spontaneous bleeding (Yang et al, Cell 151: 111-122; 2012). We now report analysis of an additional gene targeted Tmem16f allele generated in C57BL/6 ES cells. JM8 ES cell were obtained from EUCOMM, and successful Tmem16f gene targeting in intron 1 was confirmed by PCR and sequencing. Genotyping of 120 Tmem16f+/gt (+/gt) intercross progeny identified no surviving Tmem16fgt/gt (gt/gt) mice at weaning (p<0.001). However, +/gt intercrosses generated the expected Mendelian genotype ratios at both E10.5 and E17.5, with gt/gt embryo’s exhibiting no morphological abnormalities on gross or routine histologic examination. Though complete deficiency of TMEM16F is lethal in the C57BL/6J genetic background between E17.5 and birth, an F2 intercross of +/gt mice outcrossed one generation to 129x1SvJ resulted in gt/gt mice surviving to weaning, though at reduced numbers (6/75 total progeny compared to ~19 expected, p <0.002). Progeny testing of surviving gt/gt mice suggest a single autosomal dominant 129x1SvJ-associated genetic modifier. Preliminary genetic analysis of these mice appears to map this locus to the proximal region of chromosome 3. Tail bleeding times for gt/gt were >10min, whereas littermate +/gt and +/+ mice bleeding ceased at 8 ± 1 min and 6 ± 0.8 min, respectively, each significantly different than gt/gt (p<0.05). Notably, platelets from +/gt mice exhibited a trend toward reduced PS exposure, detected with FITC-labelled lactadherin, in response to PAR4 agonist peptide, whereas gt/gt mice had significantly reduced PS exposure (p < 0.05). gt/gt platelets showed a trend toward reduced PS exposure in response to A23187, as well as prolonged platelet rich plasma clotting times, and less efficient lactadherin inhibition of platelet clotting time. These data suggest the existence of a viability-determining genetic modifier of TMEM16F in the 129x1SvJ mouse strain. Identification of the responsible gene may uncover novel functions for TMEM16F and the regulation of hemostatic function. The observation of a PS exposure and hemostatic phenotype in +/gt mice also suggests the possibility that heterozygous TMEM16F mutations may subtly influence hemostasis or thrombosis in humans. Disclosures No relevant conflicts of interest to declare.

Effect of arginine supplementation of broiler breeder hens on progeny performance

Fernandes, J. I. M., Murakami, A. E., Gomes de Souza, L. M., Ospina-Rojas, I. C. and Rossi, R. M. 2014. Effect of arginine supplementation of broiler breeder hens on progeny performance. Can. J. Anim. Sci. 94: 313–321. Two experiments were conducted to determine the effects of arginine (Arg) supplementation of broiler breeder hens on the performance, carcass yield, and bone measurements of their progeny. In both experiments, the maternal diet was supplemented with five levels of digestible Arg (0.94, 1.09, 1.24, 1.39 and 1.54%). In exp. 1, a total progeny of 1050 chicks were housed in pens according to maternal diet and fed a typical diet without L-Arg supplementation. In exp. 2, a total progeny of 960 chicks were kept in pens according to maternal diet and fed diets containing supplemental L-Arg from 1.30 to 1.90% in the starter phase and from 1.15 to 1.75% in the grower phase. The data obtained in both experiments were deployed in orthogonal polynomials to allow for an analysis of variance and a regression analysis. In the starter phase, there was a quadratic effect (P<0.05) of Arg level in the maternal diet on the feed:gain ratio of the non-supplemented progeny. In the Arg-supplemented progeny, there was a quadratic effect (P<0.05) of Arg level on the feed intake and feed:gain ratio and a linear increase (P<0.05) in body weight gain, and carcass and breast yields (P<0.05). Femur length, tibia diameter, and the Seedor index of both bones increased linearly (P<0.05) in broilers fed the Arg-supplemented diet. Arg supplementation in the broiler breeder hen diets had little positive effect on the non-supplemented progeny; thus, Arg supplementation in the progeny diet is necessary to optimize performance, carcass yield, and bone quality of these hens’ progeny.