Genome-wide association study for rib eye muscle area in a Large White×Minzhu F_2 pig resource population

Rib eye muscle area(REMA) is an economically important trait and one of the main selection criteria for breeding in the swine industry. In the genome-wide association study(GWAS), the Illumina Porcine SNP60 Bead Chip containing 62 163 single nucleotide polymorphisms(SNPs) was used to genotype 557 pigs from a porcine Large White×Minzhu intercross population. The REMA(at the 5th–6th, 10th–11th and the last ribs) was measured after slaughtered at the age of(240±7) d for each animal. Association tests between REMA trait and SNPs were performed via the Genome-Wide Rapid Association using the Mixed Model and Regression-Genomic Control(GRAMMAR-GC) approach. From the Ensembl porcine database, SNP annotation was implemented using Sus scrofa Build 10.2. Thirty-three SNPs on SSC12 and 3 SNPs on SSC2 showed significant association with REMA at the last rib at the chromosome-wide significance level. None of the SNPs of REMA at the 5th–6th rib and only a few numbers of the SNPs of REMA at the 10th–11th ribs were found in this study. The Haploview V3.31 program and the Haplo.Stats R package were used to detect and visualize haplotype blocks and to analyze the association of the detected haplotype blocks with REMA at the last rib. A linkage analysis revealed that 4 haplotype blocks contained 4, 4, 2, and 4 SNPs, respectively. Annotations from pig reference genome suggested 2 genes(NOS2, NLK) in block 1(266 kb), one gene(TMIGD1) in block 2(348 kb), and one gene(MAP2K4) in block 3(453 kb). A functional analysis indicated that MYH3 and MYH13 genes are the potential genes controlling REMA at the last rib. We screened several candidate intervals and genes based on the SNPs location and the gene function, and inferred that NOS2 and NLK genes maybe the main genes of REMA at the last ribs.     

Identifying SNPs associated with birth weight and days to 100 kg traits in Yorkshire pigs based on genotyping-by-sequencing

Birth weight(BW) and days to 100 kg(D100) are important economic traits that are both affected by polygenes. However, the genetic architecture of these quantitative traits is still elusive. Genotyping-by-sequencing(GBS) data containing a large number of single nucleotide polymorphisms(SNPs) have become a powerful tool in genomic analysis. To better understand their complex genetic structure, a total of 600 Yorkshire pigs were sequenced using GBS technology. After quality control, 279 787 SNPs were generated for subsequent genome-wide association study(GWAS). A total of 30 genome-wide SNPs(P1.79 E–07) were identified for D100. Furthermore, a total of 22 and 2 suggestive SNPs(P3.57 E–06) were detected for D100 and BW, respectively. Of these, one locus located on SSC12(position: 46 226 512 bp) were evaluated to affect both BW and D100 in Yorkshire pigs, indicating the pleiotropism in different traits. Considering the function of candidate genes, two genes, NSRP1 and DOCK7, were suggested as the most promising candidate genes involved in growth traits. Thus, use of GBS is able to identify novel variants and potential candidate genes for BW and D100, and provide an opportunity for improving pig growth traits using genomic selection in pigs.     

A Method for Accuracy of Genetic Evaluation by Utilization of Canadian Genetic Evaluation Information to Improve Heilongjiang Holstein Herds

The objectives of this study were to set up a new genetic evaluation procedure to predict the breeding values of Holstein herds in Heilongjiang Province of China for milk and fat production by utilizing Canadian pedigree and genetic evaluation information and to compare the breeding values of the sires from different countries. The data used for evaluating young sires for the Chinese Holstein population consisted of records selected from 21 herds in Heilongjiang Province. The first lactation records of 2 496 daughters collected in 1989 and 2000 were analyzed. A single-trait animal model including a fixed herd-year effect, random animal and residual effects was used by utilizing Canadian pedigree and genetic evaluation information of 5126 sires released from the Canadian Dairy Network in August 2000. The BLUP procedure was used to evaluate all cattle in this study and the Estimated Breeding Values (EBV) for milk and fat production of 6697 cattle (including 673 sires and 6024 cows) were predicted. The genetic levels of the top 100 sires originated from different countries were compared. Unlike the BLUP procedure that is being used in conjunction with the single-trait sire model in Heilongjiang Province of China now, the genetic evaluation procedure used in this study not only can be used simultaneously to evaluate sires and cows but also increase the accuracy of evaluation due to using the relationships and genetic values of the Canadian evaluated sires with more daughters. The results showed that the new procedure was useful for genetic evaluation of dairy herds and the comparison of the breeding values of these sires imported from different countries showed that a significant genetic improvement has been achieved for milk production of the Heilongjiang Holstein dairy population by importing sires from foreign countries, especially from the United States due to the higher breeding values.     

Model Selection in Estimation of Covariance Functions for Growth of Angora Goats

Covariance functions have been proposed as an alternative to model longitudinal data in animal breeding because of their various merits in comparison to the classical analytical methods.In practical estimation,different models and polynomial orders fitted can influence the estimates of covariance functions and thus genetic parameters.The objective of this study was to select model for estimation of covariance functions for body weights of Angora goats at 7 time points.Covariance functions were estimated by fitting 6 random regression models with birth year,birth month,sex,age of dam,birth type,and relative birth date as fixed effects.Random effects involved were direct and maternal additive genetic,and animal and maternal permanent environmental effects with different orders of fit.Selection of model and orders of fit were carried out by likelihood ratio test and 4 types of information criteria.The results showed that model with 6 orders of polynomial fit for direct additive genetic and animal permanent environmental effects and 4 and 5 orders for maternal genetic and permanent environmental effects,respectively,were preferable for estimation of covariance functions.Models with and without maternal effects influenced the estimates of covariance functions greatly.Maternal permanent environmental effect does not explain the variation of all permanent environments,well suggesting different sources of permanent environmental effects also has large influence on covariance function estimates.     

Alterations of Alternative Splicing Patterns of Ser/Arg-Rich （SR） Genes in Response to Hormones and Stresses Treatments in Different Ecotypes of Rice （Oryza sativa）

Ser/Arg-rich (SR) genes encode proteins that play pivotal roles in both constitutive and alternative splicing of pre-mRNA. However, not much effort has been made to investigate the alternative splicing of their own pre-mRNA. In this study, we conducted comprehensive analyses of pre-mRNA splicing for 22 SR genes in three rice (Oryza sativa L.) ecotypes indica, japonica and javanica. Using different ecotypes we characterized the variations in expression and splicing patterns of rice SR genes in different tissues and at different developmental stages. In addition, we compared the divergence in expression and splicing patterns of SR genes from seedlings of different rice ecotypes in response to hormones application and environmental stresses. Our results revealed the complexity of alternative splicing of SR genes in rice. The splicing varies in different tissues, in different ecotypes, in response to stresses and hormones. Thus, our study suggested that SR genes were subjected to sophisticated alternative splicing although their encoding proteins were involved in the splicing process.     

Germplasm Evaluation,Inheritance,Molecular Marker and Mechanism of Resistance to Frog—eye Leaf Spot

Soybean frog-eye leaf spot(FLS)has been a major disease and production constralnt in the Northeast of China.The paper reported methods for identifying and evaluating resistant germplasm to the disease,at sometime listed several resistant cultivers or lines which were resistant to both dominant races(1,7race)and the mixture of over 8 races.It indicated that the resistance to race 1 or 7 were due to two different single dominant genes,respectively.And in he fied,there were more races,the resistance showed quantitative character.It was proposed the LTP,which was made according to the sizeand number of lesions could be used as an important index in selecting.Using bulked segregants analyxsis(BSA),the genetic distance between OPSO3 620 and resistant gene to race 7 wer identified 8.7cm.he results showed that the resistant cultivars had less number of stoma,closer arrangement of poliseade tissue and high cotent of wax.     

The Effects of Mx1 Locus on Immunity Tracts Components in Large White×Meishan F2 Offspring

The mouse Myxovirus resistance protein 1 (Mx1) is known to be sufficient to confer resistance to influenza viruses, and genes encoding Myxovirus resistance protein 1 (Mx1) is, therefore, an interesting candidate gene for disease resistance in farm animals. The porcine Mxl gene has already been identified and characterized based on its homology with mouse Mxl; the full-length coding region of the pig Mxl gene spans 2 545 bp (M65087) and is organized into 17 exons compared with the human ortholog mRNA. In this study, the exons 9, 10, 11 and introns 6, 9 of the porcine Mxl gene were cloned and sequenced; two SNPs were identified in exons 9, 10, 11 but none of the SNPs led to an amino acid exchange, and the other eleven variants were detected in introns 6 and 9, respectively. Differences in allele frequency among Meishan, Large White, Tibetan, Tongcheng, Huainan, and Duroc pigs were observed within intron 9, of which an A → G substitution at position 186 was detected as an Msp Ⅰ PCR-restriction fragment length polymorphism (PCR-RFLP). The association analysis using the Large White×Meishan F2 offspring suggested that the Mxl genotype was associated with variation in several immunity traits that are of interest in pig breeding. However, further investigations in more populations are needed to confirm the above concept.     

Sources for Heat-Stable Resistance to Southern Root-Knot Nematode （Meloidogyne incognita） in Solanum lycopersicum

Southern root-knot nematode （Meloidogyne incognita） is a major problem in vegetable production in China due to the expansion of plastic tunnel and solar greenhouse. Using resistant cultivars is an effective approach to control the disease. Nine genes, Mi-1 to Mi-9, have been reported and only Mi-1 has been successfully used in tomato breeding. However, Mi-1 is inactive at a temperature above 28~C. In order to identify sources for heat-stable resistance to southern root-knot nematode, 53 genotypes of tomato （Solarium spp.） were inoculated with an isolate of M. incognita in the growth chamber at 28 or 32℃ for initial screening. 28 lines had less than 25 galls and were considered as resistant candidates. The top 60% （16 in total） of resistant candidates obtained from each temperature were subject to re-evaluation at 32~C using the same nematode isolate. Three lines ZN17, ZN 48, and LA0385 showed heat-stable resistance with an average of 10 galls or less per plant. LA0385 is a wild species, while ZNI7 and ZN48 are elite breeding lines. These lines were grown in a greenhouse for two seasons, and also showed high resistance with less than 10 galls per plant. Thus they were considered as good sources for breeding resistance to southern root-knot nematode in tomato.     

The Mining of Citrus EST-SNP and Its Application in Cultivar Discrimination

Single nucleotide polymorphisms （SNPs） are the most abundant sequence variations found in plant genomes and are widely used as molecular genetic markers in cultivar identification and genetic diversity studies. The objective of this study was to identify SNP markers useful for discrimination of citrus cultivars, since large numbers of expressed sequence tags （ESTs） of sweet orange are available from the National Center for Biotechnology Information （NCBI）. We now have the opportunity to discover SNP markers suitable for determining the haplotypes with which to distinguish very closely related cultivars and to assess genetic diversity within or between related species of citrus. SNPs and small insertions/deletions （Indels） from ESTs of sweet orange and satsuma were identified by the in silico SNP discovery strategy. 55 296 EST sequences of sweet orange and 2 575 of satsuma retrieved from the NCBI repository were mined for potential SNPs. Cleaved amplified polymorphic sequences （CAPS） and sequencing approaches were used to validate putative SNPs in a sample of 30 citrus accessions. A total of 3 348 putative SNPs were identified based on the abundance of sequences and haplotype cosegregation. Of these 3 348 SNPs, the transitions, transversions and Indels ratios were 47.9, 36.1 and 16.0%, respectively. The SNPs occurred on average at a frequency of 1 per 164 bp in the coding region of citrus. 14 SNPs were randomly selected and genotyped according to 30 citrus accessions including 23 accessions of sweet orange; 11 SNPs displayed polymorphism with an average polymorphism information content （PIC） of 0.20 among 30 citrus accessions. The genetic diversity present in sweet orange was low, so the 14 SNP markers failed to discriminate different cultivars of sweet orange, but they did succeed in distinguishing accessions of inter-species of citrus. In this study, SNPs were mined from EST sequences of sweet orange and satsuma, which displayed potential capability as molecular markers to discriminate inter-species accessions of citrus. It is anticipated that these putative SNPs could be applied in citrus genetics research and breeding.     

Rapid mapping of candidate genes for cold tolerance in Oryza rufipogon Griff. by QTL-seq of seedlings

Cold stress is a major problem in rice production. To rapidly identify genes for cold tolerance in Dongxiang wild rice(DWR, Oryza rufipogon Griff.), sequencing-based bulked segregant analysis of QTL-seq method was used to resequence the extremely resistant(R) and susceptible(S) bulks of a backcross inbred lines(BILs) population(derived from Oryza sativa×O. rufipogon) and their parents. Single nucleotide polymorphisms(SNP)-index graphs and corresponding Δ(SNPindex) graphs(at 99 and 95% confidence levels) for R-and S-bulks detected a total of 2 609 candidate SNPs, including 58 candidate cold-tolerance genes. Quantitative real-time PCR analysis revealed that 5 out of the 58 candidate genes had significant differences in expression between O. sativa and O. rufipogon. Structural variation and functional annotations of the 5 candidate genes were also analyzed, and allowed us to identify 2 insertion-deletion(InDel) markers(12-7 and 12-16) that were linked with candidate genes on chromosome 12 in DWR. These results are helpful for cloning and using cold tolerance genes from common wild rice in cultivated rice.     

Genome-Wide Expression Profile of Maize Root Response to Phosphorus Deficiency Revealed by Deep Sequencing

Phosphorus （P） is one of the three primary macronutrients that are required in large amounts for plant growth and development. To better understand molecular mechanism of maize and identify relevant genes in response to phosphorus deficiency, we used Solexa/Illumina＇s digital gene expression （DGE） technology to investigate six genome-wide expression profiles of seedling roots of the low-P tolerant maize inbred line 178. DGE studies were conducted at 6, 24 and 72 h under both phosphorus deficient and sufficient conditions. Approximately 3.93 million raw reads for each sample were sequenced and 6 816 genes exhibited significant levels of differential expressions in at least one of three time points in response to P starvation. The number of genes with increased expression increased over time from 6 to 24 h, whereas genes with decreased expression were more abundant at 72 h, suggesting a gradual response process for P deficiency at different stages. Gene annotations illustrated that most of differentially expressed genes （DEGs） are involved in different cellular and molecular processes such as environmental adaptation and carbohydrate metabolism. The expression of some known genes identified in other plants, such as those involved in root architecture, P metabolism and transport were found to be altered at least two folds, indicating that the mechanisms of molecular and morphological adaptation to P starvation are conserved in plants. This study provides insight into the general molecular mechanisms underlying plant adaptation to low-P stress and thus may facilitate molecular breeding for improving P utilization in maize.     

Study of the Adult-Plant Resistance at Different Growth Stages to Stripe Rust in Wheat

Components of resistance were investigated for five adult-plant resistant (APR) wheat (Triticum aestivum) cultivars at six growth stages and two temperatures in the greenhouse, and disease progress in a field trial. Chinese cultivar Chuanyu 12 displayed high to intermediate infection type (IT) in the greenhouse but was highly resistant in the field. Weebill showed an intermediate IT in the greenhouse and also in the field. Chpaio, Tukuru and Saar, known to carry combinations of Yr18 and 2-3 additional minor genes, were highly resistant in both experiments. Greenhouse experiments indicated that the lower IT of APR cultivars initiated at tillering stage. Latent periods (LP) for APR cultivars were generally longer as the growth stage progressed. We conclude that APR to stripe rust can be best characterized in field trials although significant correlations are seen between field severity and IT and LP measured in the greenhouse.     

Genetic Parameters for Yields and Reproductive Traits of Chinese Holstein Cows in Heilongjiang Province

[Objective] This study aimed to estimate the genetic parameters for milk yield, fat percentage, fat yield and age at first freshening of Chinese Holsteins first lactation cows in Heilongjiang under frigid-temperature and temperature zone. [Method] Records of yield and reproduction from 8 868 Chinese Holstein first lactation cows from 21 herds were utilized to evaluate genetic parameters for Heilongjiangs’ Chinese Holsteins. Restricted maximum likelihood (REML) method was used with an animal model. [Result] The heritabilities were 0.16 to 0.34 for yields and fat percentage, 0.38 for reproduction (age at the first freshening), which were similar to estimates for temperate areas from similar methods, except for constituent percentage. Also, the estimates of genetic correlation were high between milk yield and fat yield (0.97), but were low between milk yield and fat percentage (-0.24), age at the first freshening (-0.29) and generally antagonistic. [Conclusion] Thus, estimates of genetic parameters in this frigid and frigid-temperate environment did not differ appreciably from those that occur in temperate dairy areas.     

Selection of reference genes for RT-qPCR analysis of Phenacoccus solenopsis (Hemiptera: Pseudococcidae) sex-dimorphic development

Mealybugs, such as Phenacoccus solenopsis, are highly sexually dimorphic. Winged adult males present such remarkable morphological differences from females that, to the untrained eye, conspecific adults of both sexes of P. solenopsis may be considered as two different insect species. A method to investigate sex-dimorphic mechanisms is by evaluating gene expression using RT-qPCR. However, the accuracy and consistency of this technique depend on the reference gene(s) selected. In this study, we analyzed the expression of 10 candidate reference genes in male and female P. solenopsis at different development stages, using common algorithms including the ?Ct method, NormFinder, geNorm, BestKeeper, and a web-based analysis tool, RefFinder. The results showed that EF1-β, RP-L32 and RP-18 S were selected as the most stable genes by both the ?Ct method and NormFinder; TUB-α was the most stable gene identified by BestKeeper; and RP-L40 and RP-L32 were the most stable genes ranked by geNorm. RefFinder, a comprehensive analysis software, ranked the ten genes and determined EF1-β and RP-L32 as the most suitable reference genes for the various developmental stages in male and female P. solenopsis. Furthermore, the two most suitable reference genes were validated by examining expression of the juvenile hormone acid O-methytransferase(JHAMT) gene. Results of the validation portion of the study showed that JHAMT expression was sex-biased towards males and exhibited a dynamic and classic expression pattern among the P. solenopsis developmental stages. The results can help further our knowledge on the molecular mechanisms underlying sexual dimorphic development in P. solenopsis.