Enhancing host immunity is an effective way to reduce morbidity in chickens.  Heterophil to lymphocyte ratio (H/L) is associated with host disease resistance in birds.  Chickens with different H/L levels show different disease resistances.  However, the utility of the H/L as an indicator of immune function needs to be further analyzed.  In this study, a H/L directional breeding chicken line (Jingxing yellow chicken) was constructed, which has been bred for 12 generations.  We compared the function of heterophils, and combined statistical analysis to explore the candidate genes and pathways related to H/L.  The oxidative burst function of the heterophils isolated from the H/L selection line (G12) was increased (P=0.044) compared to the non-selection line (NS).  The 22.44 Mb genomic regions which annotated 300 protein-coding genes were selected in the genome of G9 (n=92) compared to NS (n=92) based on a genome-wide selective sweep.  Several selective regions were identified containing genes like interferon induced with helicase C domain 1 (IFIH1) and moesin (MSN) associated with the intracellular receptor signaling pathway, C–C motif chemokine receptor 6 (CCR6), dipeptidyl peptidase 4 (DPP4) and hemolytic complement (HC) associated with the negative regulation of leukocyte chemotaxis and tight junction protein 1 (TJP1) associated with actin cytoskeleton organization.  In addition, 45 genome-wide significant indels containing 29 protein-coding genes were also identified as associated with the H/L based on genome-wide association study (GWAS).  The expression of protein tyrosine phosphatase non-receptor type 5 (PTPN5) (r=0.75, P=0.033) and oxysterol binding protein like 5 (OSBPL5) (r=0.89, P=0.0027) were positively correlated with H/L.  Compared to the high H/L group, the expressions of PTPN5 and OSBPL5 were decreased (P<0.05) in the low H/L group of Beijing you chicken.  The A/A allelic frequency of indel 5_13108985 (P=3.85E–06) within OSBPL5 gradually increased from the NS to G5 and G9, and the individuals with A/A exhibited lower H/L than individuals with heterozygote A/ATCT (P=4.28E–04) and homozygous ATCT/ATCT (P=3.40E–05).  Above results indicated oxidative burst function of heterophils were enhanced, and 22.44 Mb genomic regions were selected with the directional selection of H/L.  In addition, PTPN5 and OSBPL5 genes were identified as H/L-related candidate genes.  These findings revealed the complex genetic mechanism of H/L related to immunity and will allow selection for improving chicken immunity based on the H/L

Photosynthesis occurs mainly in chloroplasts, whose development is regulated by proteins encoded by nuclear genes.  Among them, pentapeptide repeat (PPR) proteins participate in organelle RNA editing.  Although there are more than 450 members of the PPR protein family in rice, only a few affect RNA editing in rice chloroplasts.  Gene editing technology has created new rice germplasm and mutants, which could be used for rice breeding and gene function study.  This study evaluated the functions of OsPPR9 in chloroplast RNA editing in rice.  The osppr9 mutants were obtained by CRISPR/Cas9, which showed yellowing leaves and a lethal phenotype, with suppressed expression of genes associated with chloroplast development and accumulation of photosynthetic-related proteins.  In addition, loss of OsPPR9 protein function reduces the editing efficiency of rps8-C182, rpoC2-C4106, rps14-C80, and ndhB-C611 RNA editing sites, which affects chloroplast growth and development in rice.  Our data showed that OsPPR9 is highly expressed in rice leaves and encodes a DYW-PPR protein localized in chloroplasts.  Besides, the OsPPR9 protein was shown to interact with OsMORF2 and OsMORF9.  Together, our findings provide insights into the role of the PPR protein in regulating chloroplast development in rice. 

The red coloring of pear fruits is mainly caused by anthocyanin accumulation.  Red sport, represented by the green pear cultivar ‘Bartlett’ (BL) and the red-skinned derivative ‘Max Red Bartlett’ (MRB), is an ideal material for studying the molecular mechanism of anthocyanin accumulation in pear.  Genetic analysis has previously revealed a quantitative trait locus (QTL) associated with red skin color in MRB.  However, the key gene in the QTL and the associated regulatory mechanism remain unknown.  In the present study, transcriptomic and methylomic analyses were performed using pear skin for comparisons between BL and MRB.  These analyses revealed differential PcHY5 DNA methylation levels between the two cultivars; MRB had lower PcHY5 methylation than BL during fruit development, and PcHY5 was more highly expressed in MRB than in BL.  These results indicated that PcHY5 is involved in the variations in skin color between BL and MRB.  We further used dual luciferase assays to verify that PcHY5 activates the promoters of the anthocyanin biosynthesis and transport genes PcUFGT, PcGST, PcMYB10 and PcMYB114, confirming that PcHY5 not only regulates anthocyanin biosynthesis but also anthocyanin transport.  Furthermore, we analyzed a key differentially methylated site between MRB and BL, and found that it was located in an intronic region of PcHY5.  The lower methylation levels in this PcHY5 intron in MRB were associated with red fruit color during development, whereas the higher methylation levels at the same site in BL were associated with green fruit color.  Based on the differential expression and methylation patterns in PcHY5 and gene functional verification, we hypothesize that PcHY5, which is regulated by methylation levels, affects anthocyanin biosynthesis and transport to cause the variations in skin color between BL and MRB.

Salmonella is one of the most common food-borne pathogens and its resistance in chicken can be improved through genetic selection.  The heterophils/lymphocytes (H/L) ratio in the blood reflects the immune system status of chicken.  We compared the genome data and spleen transcriptomes between the H/L ratio-selected and non-selected chickens, after Salmonella infection, aiming to identify the key genes participating in the antibacterial activity in the spleen.  The results revealed that, the selected population had stronger (P<0.05) liver resistance to Salmonella typhimurium (ST) than the non-selected population.  In the selected and non-selected lines, the identified differentiation genes encode proteins involved in biological processes or metabolic pathways that included the TGF-beta signaling pathway, FoxO signaling pathway, and Salmonella infection pathway.  The results of the analysis of all identified differentially expressed genes (DEGs) of spleen revealed that the G protein-coupled receptor (GPCR) and insulin-like growth factor (IGF-I) signaling pathways were involved in the Salmonella infection pathway.  Integrated analysis of DEGs and F_ST (fixation index), identified candidate genes involved in Salmonella infection pathway, such as GPR39, NTRK2, and ANXA1.  The extensive genomic changes highlight the polygenic genetic of the immune response in these chicken populations.  Numerous genes related to the immune performance are differentially expressed in the selected and non-selected lines and the selected lines has a higher resistance to Salmonella. 

Winter jujube (Ziziphus jujuba ‘Dongzao’) is an excellent late maturing variety of fresh-eating jujube in China.  Fruit texture is an important indicator of sensory quality.  To investigate the correlations among texture indices and establish an evaluation system for winter jujube texture, we used the TMS-Touch instrument to perform a texture profile analysis (TPA) on 1 150 winter jujubes from three major producing areas in China.  Eight indices and their best-fit distribution were obtained, including fracture (Pearson), hardness (InvGauss), adhesive force (Weibull), adhesiveness (LogLogistic), cohesiveness (LogLogistic), springiness (BetaGeneral), gumminess (InvGauss), and chewiness (InvGauss).  Based on the best-fit distribution curves, each index was divided into five grades (lower, low, medium, high and higher) by the 10th, 30th, 70th and 90th percentiles.  Among the texture indices, 82% of the correlation coefficients were highly significant (P<0.01); meanwhile, chewiness was significantly (P<0.01) and positively correlated with springiness and gumminess, of which the correlation coefficients were up to 0.8692 and 0.8096, respectively.  However, adhesiveness was significantly (P<0.01) and negatively related to adhesive force with a correlation coefficient of –0.7569.  Among hardness, cohesiveness, springiness, gumminess, and chewiness, each index could be well fitted by a multiple linear regression with the remaining four indices, with the coefficients above 0.94 and the mean fitting error and mean prediction error lower than 10%.  A comprehensive evaluation model was consequently established based on factor analysis to evaluate the texture quality of winter jujube.  The results demonstrated that winter jujube with higher comprehensive scores generally exhibited higher springiness and chewiness, but had lower adhesive force and adhesiveness.  We used factor analysis and clustering analysis to divide the eight studied texture into four groups (cohesive factor, adhesive-soft factor, tough-hard factor, and crispness factor), whose representative indices were springiness, adhesiveness, hardness, and fracture, respectively.  Overall, this study investigated the variation in each index of winter jujube texture, explored the association among these indices, screened the representative indices, and established a texture evaluation system.  The results provide a methodological basis and technical support for evaluating winter jujube texture.