Cotton (Gossypium. spp) stem trichomes and seed fibers are unicellular epidermal hairs regulated by relevant molecular networks. Genetic analysis of the glabrous stem landrace (Palmeri37) of Gossypium hirsutum L. has pinpointed GL2-interacting-repressor 1 (GhGIR1D), featuring a RING-like zinc finger domain, as the candidate gene underlying the stem glabrous trait. Overexpressing and silencing experiments confirm GhGIR1D as a negative regulator specifically for stem trichome initiation, without influencing leaf trichome or seed fiber. High endogenous expression of GhGIR1D is associated with a SNP-573 T/G variation within the promoter region, and GhGIR1D Hap I confers the absence of stem trichomes. The homoeologous protein, GhGIR1A, inhibits trichome and fuzz fiber initiation by disrupting the GhHD1D-GhHOX3A module (a complex of 2 Homeodomain-Leucine Zipper IV transcription factors, HD-ZIP IV), and repressing downstream transcription of GhRDL1. Trichome density was enhanced in GhHD1A overexpression transgenic lines but reduced in ghhd1a mutants, demonstrating the positive regulatory role of GhHD1A on stem trichome initiation. GhHD1 displays distinct effects on stem trichome and fuzz fiber initiation due to its tissue-specific dosage. In the ghhd1a mutants, repression of GhGIR1D restores the wild-type pubescent phenotype, suggesting the presence of a potential negative feedback loop between GhGIR1D and GhHD1A, or that GhGIR1D and GhHD1A may function within the same regulatory pathway with opposite functions in regulation of trichome development. These findings enhance our comprehension of the GhGIR1-GhHD1-GhHOX3 interaction module in epidermal hair initiation and development.

With the continuous expansion of the planting area of genetically modified (GM) crops, the demand for efficient and comprehensive monitoring systems is becoming increasingly urgent. To establish a method suitable for large-scale monitoring of genetically modified rapeseed planting, beehives were strategically deployed at specific locations around genetically modified rapeseed fields, and the TaqMan quantitative PCR (qPCR) method was used to detect and analyze the genetically modified components in the rapeseed pollen collected by bees. The results demonstrated that the average Ct values for the CaMV35S promoter, Bar gene, NPTII gene, and HPT gene in the pollen of each hive were 27.91, 29.58, 31.49, and 31.97, respectively. The average ΔCt values for these four genes in hive pollen from 100 to 200 m were − 0.35, 1.66, 2.58, and 5.06, respectively, which were significantly lower than those from 300 to 1100 m (2.85, 4.01, 6.66, and 5.63). The results of this study have demonstrated the feasibility of using pollen collected by bees for large-scale detection of genetically modified rapeseed plants. This early warning model for GM crop spread based on bee pollination provides an efficient and practical solution for monitoring and managing genetically modified crops.

As a country with abundant genetic resources of pigs, the domestication history of pigs in China and the adaptive evolution of Chinese pig breeds at different latitudes have rarely been elucidated at the genome-wide level. To fill this gap, we first assembled a high-quality chromosome-level genome of the Chenghua pig and used it as a benchmark to analyse the genomes of 272 samples from three genera of three continents. The divergence of the three species belonging to three genera, Phacochoerus africanus , Potamochoerus porcus , and Sus scrofa , was assessed. The introgression of pig breeds redefined that the migration routes were basically from southern China to central and southwestern China, then spread to eastern China, arrived in northern China, and finally reached Europe. The domestication of pigs in China occurred -12,000 years ago, earlier than the available Chinese archaeological domestication evidence. In addition, FBN1 and NR6A1 were identified in our study as candidate genes related to extreme skin thickness differences in Eurasian pig breeds and adaptive evolution at different latitudes in Chinese pig breeds, respectively. Our study provides a new resource for the pig genomic pool and refines our understanding of pig genetic diversity, domestication, migration, and adaptive evolution at different latitudes.

8029AB is a newly bred dominant genic male sterility （DGMS） two-type line in Brassica napus,whose fertility is controlled by one gene with three different alleles （BnMS5 a /BnMS5 c /BnMS5 e ）.It can be used in a three-line approach for the seed production of rapeseed hybrids.In order to promote its application,functional markers should be developed to accurately identify multiple alleles （BnMS5 a /BnMS5 c /BnMS5 e ）.In our study,15pairs of SNP markers were designed according to the sequence of BnMS5 a ,BnMS5 c and BnMS5 e .Three of them were successfully identified and validated.The SNP marker SNPMS5 a -5 was tightly linked to BnMS5 a ,which could amplify a 200 bp band in fertile individuals with BnMS5 a .SNPMS5 c -3 could be used as the molecular marker of BnMS5 c through amplifying a 140 bp band.SNPMS5 e -1 which was tightly linked to BnMS5 e could amplify one band of 150 bp in plants with BnMS5 e .These SNP markers could predict the presence of BnMS5 a /BnMS5 c /BnMS5 e effectively.Consequently,marker-assisted selection can be carried out in the early generation to shorten the breeding period and improve the breeding efficiency,so as to realize large-scale commercial production of DGMS three-line hybrid seed in rapeseed.

Different anatomic locations of the body skin dermis come from different origins, and its positional hereditary information can be maintained in adults, while highly resolvable cellular specialization is less well characterized in different anatomical regions. Pig is regarded as excellent model for human research in view of its similar physiology to human. In this study, we performed single-cell RNA sequencing of six different anatomical skin regions from the Chenghua pig with superior skin thickness trait. We obtained 215,274 cells, representing seven cell types, among which we primarily characterized the heterogeneity of smooth muscle cells, endothelial cells and fibroblasts. We identified several phenotypes of smooth muscle cell and endothelial cell and presented genes expression of pathways such as the immune response in different skin regions. By comparing differentially expressed fibroblast genes among different skin regions, we considered TNN, COL11A1, and INHBA as candidate genes for facilitating ECM accumulation. These findings of heterogeneity in the main three cell types from different anatomic skin sites will contribute to a better understanding of hereditary information and places the potential focus on skin generation, transmission and transplantation, paving the foundation for human skin priming.

Background Skin development is a complex process that is influenced by many factors. Pig skin is used as an ideal material for xenografts because it is more anatomically and physiologically similar to human skin. It has been shown that the skin development of different pig breeds is different, and some Chinese pig breeds have the characteristics of skin thickness and facial skin folds, but the specific regulatory mechanism of this skin development is not yet clear. Methods In this study, the facial skin of Chenghua sows in the four developmental stages of postnatal Day 3 (D3) , Day 90 (D90) , Day 180 (D180), and Year 3 (Y3) were used as experimental materials, and RNA sequencing (RNA–seq) analysis was used to explore the changes in RNA expression in skin development at the four developmental stages, determine the differentially expressed messenger RNAs (mRNAs), long noncoding RNAs (lncRNAs), microRNAs (miRNAs), and circular RNAs (circRNAs), and perform functional analysis of related genes by Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. Results A pairwise comparison of the four developmental stages identified several differentially expressed genes (DEGs) and found that the number of differentially expressed RNAs (DE RNAs) increased with increasing developmental time intervals. Elastin (ELN) is an important component of the skin. Its content affects the relaxation of the epidermis and dermal connection, and its expression is continuously downregulated during the four developmental stages. The functions of DEGs at different developmental stages were examined by performing GO and KEGG analyses, and the GO terms and enrichment pathways of mRNAs, lncRNAs, miRNAs, and circRNAs highly overlapped, among which the PPAR signaling pathway, a classical pathway for skin development, was enriched by DEGs of D3 vs. D180, D90 vs. D180 and D180 vs. Y3. In addition, we constructed lncRNA-miRNA-mRNA and circRNA-miRNA interaction networks and found genes that may be associated with skin development, but their interactions need further study. Conclusions We identified a number of genes associated with skin development, performed functional analyses on some important DEGs and constructed interaction networks that facilitate further studies of skin development.

猪瘟(CSFV)和猪繁殖与呼吸综合征(PRRSV)是危害我国养猪业的两类重要的传染性疾病,为了评估四川部分地区规模化养殖场猪瘟和猪繁殖与呼吸综合征疫苗免疫效果,本研究采用酶联免疫方法(ELISA)对四川成都、广元、攀枝花、内江四个猪场送检的213份猪血清样本进行了CSFV和PRRSV抗体检测.结果表明:有168份血清CSFV免疫抗体阳性,阳性率为78.87％,四个猪场中阳性率最高为89.80％,最低为63.93％;162份PRRSV抗体阳性,阳性率为76.06％,四个猪场中阳性率最高为93.15％,最低为57.38％.按不同猪群统计显示,母猪及公猪群体的猪瘟疫苗免疫效果较好,仔猪群体具有较好的母源抗体,而保育猪猪瘟免疫抗体数据较差,提示仔猪猪瘟的免疫程序及免疫剂量需要进行调整.猪繁殖与呼吸综合征免疫抗体中母猪免疫抗体效果差异明显,提示部分养殖场母猪的免疫程序及免疫剂量需进行调整.本研究结果可以为四川地区规模化养殖场CSFV与PRRSV防控提供技术支撑.

The 8029AB line is a dominant genic male sterility (DGMS) two-type line in Brassica napus L., which can be used in a three-line approach for the seed production of rapeseed hybrids. Genetic analyses have demonstrated that the sterility of 8029A is controlled by a single dominant nuclear gene (BnMS5e) interacting with one recessive gene (BnMS5c). Six pairs of penta-primer amplification refractory mutation system (PARMS) markers were designed according to the sequence of BnMS5a, BnMS5c and BnMS5e. Two pairs of these PARMS markers were successfully identified and validated. The PARMS markers MS5-1Fc/MS5-1Ft/MS5-1R12 could distinguish BnMS5c from BnMS5a/BnMS5e, and the PARMS markers MS5-2Ft/MS5-2Fa/MS5-1R12 could genotype BnMS5a and BnMS5c/BnMS5e. The combination of these two pairs of PARMS markers could be used to identify the presence or absence of BnMS5a/BnMS5c/BnMS5e effectively. Consequently, marker-assisted selection can be carried out in the early generation to shorten the breeding period and improve the breeding efficiency.

Objective This work was to determine coat inheritance and evaluate production performance for crossbred pigs from Berkshire×Chenghua (BC) compared with Chinese indigenous Chenghua (CH) pigs. Methods The coat color phenotypes were recorded for more than 16,000 pigs, and the genotypes of melanocortin 1 receptor (MCIR) gene were identified by sequencing. The reproductive performance of 927 crossbred BC F4 gilts and 320 purebred CH gilts was recorded. Sixty pigs of each breed were randomly selected at approximately 60 days of age to determine growth performance during fattening period, which lasted for 150 days for BC pigs and 240 days for CH pigs. At the end of the fattening period, 30 pigs of each breed were slaughtered to determine carcass composition and meat quality. Results The coat color of BC pigs exhibits a “dominant black” hereditary pattern, and all piglets derived from boars or sows genotyped ED1ED1 homozygous for MC1R gene showed a uniform black coat phenotype. The BC F4 gilts displayed a good reproductive performance, showing a higher litter and tear size and were heavier at farrowing litter and at weaning litter than the CH gilts, but they reached puberty later than the CH gilts. BC F4 pigs exhibited improved growth and carcass characteristics with a higher average daily live weight gain, lower feed-to-gain ratio, and higher carcass lean meat rate than CH pigs. Like CH pigs, BC F4 pigs produced superior meat-quality characteristics, showing ideal pH and meat-color values, high intramuscular fat content and water-holding capacity, and acceptable muscle-fiber parameters. C18:1, C16:0, C18:0, and C18:2 were the main fatty acids in M. longissimus lumborum in the two breeds, and a remarkably high polyunsaturated/saturated fatty acid ratio of ~0.39 was observed in the BC F4 pigs. Conclusion The BC F4 pigs exhibit a uniform black coat pattern and acceptable total production performance.

Skin thickness is closely related to the appearance of human skin, such as sagging and wrinkling, which primarily depends on the level of collagen I synthesized by fibroblasts in the dermal layer. To explore the underlying genetics of the development of skin thickness, we used the indigenous Chinese Chenghua pigs, considered to have superior skin thickness, as model animals. We first performed whole transcriptome sequencing analysis to identify significant skin morphological differences between Chenghua pigs and Large White pigs and obtained some differentially expressed coding RNAs (454 mRNAs) and noncoding RNAs (612 circRNAs, 188 miRNAs, and 19 lncRNAs); moreover, some competing endogenous RNA (ceRNA) networks were constructed. Interestingly, we then identified a circRNA, namely circ0044633, which plays an important role in promoting fibroblast prolif-eration along with myofibroblast transition and collagen I synthesis by sponging miR-23b and regulating CADM3 and MAP4K4 expression via activation of the downstream AKT and ERK pathways in vitro. Furthermore, over-expression of circ004463 increased the mouse skin thickness and collagen I content in vivo. These results revealed a whole transcript profile of skin tissue and identified an important circ0044633-miR-23b-CADM3/MAP4K4 axis related to fibroblast proliferation and collagen I synthesis during the development of skin thickness.

成华猪是我国著名地方猪种,是成都平原土生土长的黑毛猪,具有肉质优良、皮厚、抗逆性强等优势性状品种特征,是四川省发展特色生猪产业、擦亮"川猪"金字招牌的重要战略遗传资源.本文从成华猪遗传资源保护体系建立、种质特征遗传评价、天府黑猪新品种培育及推广利用等方面现状予以分析,并就下阶段成华猪遗传资源保护及利用提出建议.

2021年2月至7月期间,四川某规模猪场保育猪和育肥猪出现疫情,表现为咳嗽、喘气、渐进性消瘦,部分猪关节肿、育肥猪有呼吸困难、咳嗽、零星死亡等症状.猪场通过抗生素防治效果不好.我们通过现场诊断、剖检观察结合实验室全面诊断,确诊为圆环病毒继发支原体和副猪嗜血杆菌,并采取了相应控制方案,现将诊治过程总结如下.

Skin thickness is closely related to the appearance of human skin, such as sagging and wrinkling, which primarily depends on the level of collagen I synthesized by dermal fibroblasts (DFs). Small extracellular vesicles (SEVs), especially those derived from human DFs (HDFs), are crucial orchestrators in shaping physiological and pathological development of skin. However, the limited supply of human skin prevents the production of a large amount of HDFs-SEVs, and pig skin is used as a model of human skin. In this study, SEVs derived from DFs of Chenghua pigs (CH-SEVs), considered to have superior skin thickness, and Large White pigs (LW-SEVs) were collected to compare their effects on DFs and skin tissue. Our results showed that, compared with LW-SEVs, CH-SEVs more effectively promoted fibroblast proliferation, migration, collagen synthesis and contraction; in addition, in mouse model injected with both SEVs, compared with LW-SEVs, CH-SEVs increased the skin thickness and collagen I content more effectively. Some differentially expressed miRNAs and proteins were found between CH-SEVs and LW-SEVs by small RNA-seq and LC–MS/MS analysis. Interestingly, we identified that CH-SEVs were enriched in miRNA-218 and ITGBL1 protein, which played important roles in promoting fibroblast activity via activation of the downstream TGFβ1-SMAD2/3 pathway in vitro. Furthermore, overexpression of miRNA-218 and ITGBL1 protein increased the thickness and collagen I content of mouse skin in vivo. These results indicate that CH-SEVs can effectively stimulate fibroblast activity and promote skin development and thus have the potential to protect against and repair skin damage. Graphical Abstract

New genes (or lineage-specific genes) can facilitate functional innovations. MALE STERILITY 5 (MS5) in Brassica napus is a fertility-related new gene, which has two wild-type alleles (BnMS5(a) and BnMS5(C)) and two mutant alleles (BnMS5(b) and BnMS5(d)) that could induce male sterility. Here, we studied the history and functional evolution of MS5 homologs in plants by phylogenetic analysis and molecular genetic experiments. We identified 727 MS5 homologs and found that they define a Brassicaceae-specific gene family that has expanded partly via multiple tandem gene duplications and also probably transpositions. The MS5 in B. napus is inherited from a basic diploid ancestor of B. rapa. Molecular genetic experiments indicate that BnMS5(a) and BnMS5(C) are functionally distinct in B. napus and that BnMS5(d) can inhibit BnMS5(a) in B. napus in a dosage-dependent manner. The BnMS5(a) protein can move in coordination with meiotic telomeres and interact with the nuclear envelope protein SUN1, with a possible crucial role in meiotic chromosome behavior. In summary, BnMS5 belongs to a Brassicaceae-specific new gene family, and has gained a novel function that is essential for male fertility in B. napus through neofunctionalization that has likely occurred since the origin of B. rapa.

转基因作物的分子特征为转基因的安全性评估和后续监测奠定了基础.由于靶点的特异性,以聚合酶链反应(PCR)为基础的传统分子特征鉴定方法不能全面检测外来基因的非预期插入、载体骨架残留及未知的转基因事件.更多新的育种技术产生的基因修饰作物也对传统分子特征鉴定方法提出了挑战.下一代测序技术,可以克服基于PCR的方法的某些局限性,提供快速、全面分子特征数据.本文综述了T-DNA整合的复杂机制,传统分子特征鉴定方法的局限性,高通量测序方法在转基因分子特征安全评价中的应用、遇到的挑战及应用策略,为转基因的安全评价工作提供借鉴.

BACKGROUND:Cotton stem trichomes and seed fibers are each single celled structures formed by protrusions of epidermal cells, and were found sharing the overlapping molecular mechanism. Compared with fibers, cotton stem trichomes are more easily observed, but the molecular mechanisms underlying their development are still poorly understood.RESULTS:In this study, Gossypium hirsutum (Gh) and G. barbadense (Gb) were found to differ greatly in percentages of varieties/accessions with glabrous stems and in trichome density, length, and number per trichopore. Gh varieties normally had long singular and clustered trichomes, while Gb varieties had short clustered trichomes. Genetic mapping using five F2 populations from crosses between glabrous varieties and those with different types of stem trichomes revealed that much variation among stem trichome phenotypes could be accounted for by different combinations of genes/alleles on Chr. 06 and Chr. 24. The twenty- six F1 generations from crosses between varieties with different types of trichomes had varied phenotypes, further suggesting that the trichomes of tetraploid cotton were controlled by different genes/alleles. Compared to modern varieties, a greater proportion of Gh wild accessions were glabrous or had shorter and denser trichomes; whereas a smaller proportion of Gb primitive accessions had glabrous stems. A close correlation between fuzz fiber number and stem trichome density was observed in both Gh and Gb primitive accessions and modern varieties.CONCLUSION:Based on these findings, we hypothesize that stem trichomes evolved in parallel with seed fibers during the domestication of cultivated tetraploid cotton. In addition, the current results illustrated that stem trichome can be used as a morphological index of fiber quality in cotton conventional breeding.

With the successful completion of genomic sequencing for Brassica napus, identification of novel genes, determination of functions performed by genes, and exploring the molecular mechanisms underlying important agronomic traits were challenged. Mutagenesis-based functional genomics techniques including chemical, physical, and insertional mutagenesis have been used successfully in the functional characterization of genes. However, these techniques had their disadvantages and inherent limitations for allopolyploid Brassica napus, which contained a large number of homologous and redundant genes. Long intron-spliced hairpin RNA (ihpRNA) constructs which contained inverted repeats of the target gene separated by an intron, had been shown to be very effective in triggering RNAi in plants. In the present study, the genome-wide long ihpRNA library of B. napus was constructed with the rolling circle amplification (RCA)-mediated technology. Using the phytoene desaturase (PDS) gene as a target control, it was shown that the RCA-mediated long ihpRNA construct was significantly effective in triggering gene silence in B. napus. Subsequently, the resultant long ihpRNA library was transformed into B. napus to produce corresponding RNAi mutants. Among the obtained transgenic ihpRNA population of B. napus, five ihpRNA lines with observable mutant phenotypes were acquired including alterations in the floral model and the stamen development. The target genes could be quickly identified using specific primers. These results showed that the RCA-mediated ihpRNA construction method was effective for the genome-wide long ihpRNA library of B. napus, therefore providing a platform for study of functional genomics in allopolyploid B. napus.

异源四倍体油菜含有大量多拷贝基因及冗余基因,难以通过T-DNA插入、物理及化学诱变等常规突变体库创制方法发掘功能基因.为开发适应于油菜简单高效的突变体库创制方法,本研究通过滚环复制方法构建了甘蓝型油菜的花序lhRNAi文库,并对这种新型的干扰文库进行质量评估;然后将该花序lhRNAi文库对甘蓝型油菜进行遗传转化,获得763株T 0植株,从中鉴定分离出74株具有可见表型变异的花发育相关突变体,包括花瓣减少、形状异常,柱头卷曲,雄蕊退化萎缩,雄性不育、死蕾或花蕾闭合等.说明通过滚环复制介导的lhRNAi文库的构建方法可以成功应用到油菜中,这将为油菜花序发育相关基因的功能研究提供重要的研究平台.

Plant stems are involved in supporting the entire plant body, thus having an important effect on the yield of oilseed rape. The current understanding of the mechanism of stem development in oilseed rape is limited. In this study, a detailed comparative analysis of mRNA expression levels in Brassica napus between strong- (sample numbers 7110, 7031, 7029, 7095, and 7109) and weak-stemmed (sample numbers 7003, 7088, and 7123) was performed using RNA sequencing. A total of 1531 significantly differentially expressed genes were identified, including genes involved in cellulose and lignin biosynthesis, cell wall-binding proteins, and hormone biosynthesis and response genes. These results provide valuable insights into the molecular basis of stem development at the transcriptional level and will be helpful for revealing the regulatory mechanisms of stem growth in B. napus.

Chromosome dynamics during meiotic prophase I are associated with a series of major events such as chromosomal reorganization and condensation, pairing/synapsis and recombination of the homologs, and chromosome movements at the nuclear envelope (NE). The NE is the barrier separating the nucleus from the cytoplasm and thus plays a central role in NE-associated chromosomal movements during meiosis. Previous studies have shown in various species that NE-linked chromosome dynamics are actually driven by the cytoskeleton. The linker of nucleoskeleton and cytoskeleton (LINC) complexes are important constituents of the NE that facilitate in the transfer of cytoskeletal forces across the NE to individual chromosomes. The LINCs consist of the inner and outer NE proteins Sad1/UNC-84 (SUN), and Klarsicht/Anc-1/Syne (KASH) domain proteins. Meiosis-specific adaptations of the LINC components and unique modifications of the NE are required during chromosomal movements. Nonetheless, the actual role of the NE in chromosomic dynamic movements in plants remains elusive. This review summarizes the findings of recent studies on meiosis-specific constituents and modifications of the NE and corresponding nucleoplasmic/cytoplasmic adaptors being involved in NE-associated movement of meiotic chromosomes, as well as describes the potential molecular network of transferring cytoplasm-derived forces into meiotic chromosomes in model organisms. It helps to gain a better understanding of the NE-associated meiotic chromosomal movements in plants.