Construction and Partial Biological Characteristics Analysis of Listeriolysin S llsB Gene Deletion Strain

In order to analyze the effect of listeriolysin S (LLS) llsB gene deletion on the biological characteristics of Listeria monocytogenes (LM),this study used homologous recombination to construct the llsB gene deletion strain LM90-ΔllsB,and the biological characteristics of growth characteristics,median lethal dose (LD₅₀) and organ-borne bacteria were studied in healthy Kunming male mice at 8 weeks old and weighing 40 g±5 g.The llsB gene deletion strain was successfully constructed,and the deletion strain had good genetic stability through continuous passage to 20 generations in vitro.Based on its growth curve examination,we found that the growth rate of the mutant strain was slightly higher than that of the parent strain.The results of mice infection test showed that the LD₅₀ of the parent strain and the deletion strain were 10^6.17 and 10^6.50 CFU,respectively.Compared with the parent strain,the amount of bacteria load of the deletion strain in the liver and spleen of the mice was extremely significantly decreased (P<0.01),the results showed that the infection ability of the mutant strain on mice was obviously weakened.No Listeria monocytogenes was detected in the brain.The results suggested that llsB gene might have direct or indirect regulatory effect on some biological characteristics of LM90,and it would provide a theoretical basis for further understanding of the pathogenic mechanism of LLS,prevention and control of listeriosis.     

Intestinal Microbiota in Large Yellow Croaker,Larimichthys crocea,at Different Ages

The polymerase chain reaction–denaturing gradient gel electrophoresis (PCR‐DGGE) of 16S ribosomal RNA gene was used to investigate bacterial communities in the intestines of large yellow croaker at six different ages (12 d, 18 d, 26 d, 40 d, 3 mo, and 1 yr old) as well as within the corresponding feed and culture water. In addition, Illumina Miseq sequencing was utilized to compare intestinal microbiota between 12‐d‐old and 1‐yr‐old individuals. PCR‐DGGE results revealed that the culture water had the highest bacterial diversity, followed by the feed, while the intestines had the lowest diversity. The intestinal microbiota at six ages changed severely; however, the change did not follow any trend. The large yellow croaker intestines harbored specific bacterial communities that differed from those in both feed and water. Illumina Miseq sequencing results revealed that the diversity of intestinal bacteria in 12‐d‐old fish was higher than that in 1‐yr‐old fish, and the bacterial composition differed significantly between them. γ‐Proteobacteria and Pseudoalteromonas supplied the most abundant phylum and genus in the 12‐d‐old fish intestine. However, in the 1‐yr‐old fish intestine, Firmicutes and Clostridium were the most dominant, respectively. The study may contribute to a better understanding of gut microbiota and dynamics of the large yellow croaker and the relationship with their surrounding environment.     

Identification,characterization and full-length sequence analysis of a novel endornavirus in common sunflower (Helianthus annuus L.)

To identify the possible quarantine viruses in seven common sunflower varieties imported from the United States of America and the Netherlands, we tested total RNAs extracted from the leaf tissues using next-generation sequencing of small RNAs. After analysis of small RNA sequencing data, no any quarantine virus was found, but a double-stranded RNA(dsRNA) molecule showing typical genomic features of endornavirus was detected in two varieties, X3939 and SH1108. Full-length sequence and phylogenetic analysis showed that it is a novel endornavirus, temporarily named as Helianthus annuus alphaendornavirus(HaEV). Its full genome corresponds to a 14 662-bp dsRNA segment, including a 21-nt 5′ untranslated region(UTR), 3' UTR ending with the unique sequence CCCCCCCC and lacking a poly(A) tail. An open reading frame(ORF) that encodes a deduced 4 867 amino acids(aa) polyprotein with three domains: RdRP, Hel and UGT(UDP-glycosyltransferase). HaEV mainly distributed in the cytoplasm but less in the nucleus of leaf cells by fluorescence in situ hybridization(FISH) experiment. This virus has a high seed infection rate in the five varieties, X3907, X3939, A231, SH1108 and SR1320. To our knowledge, this is the first report about the virus of the family Endornaviridae in the common sunflower.     

魏可葡萄气生根发育的转录组分析

葡萄气生根是在高温高湿环境下老蔓上发育形成的一种特殊根系类型,发育程度较低,具有分化潜力。通过技术手段诱导魏可葡萄当年生结果枝茎节处萌发气生根,以同一结果母枝未处理茎节为对照进行根系转录基因表达情况分析,结果表明,诱导气生根发育过程中有3 989个基因表达量上升,2 830个基因表达量下调。采用GO功能分类,可将注释的基因划分为40个功能类别,包括催化活性、结合和代谢过程等。部分根系关键基因表达结果表明,生长素、赤霉素、脱落酸在内的所有植物激素都参与了调控过程,结合生长素下游相应基因分析,内切葡聚糖酶基因、PRE基因和高亲和力硝酸盐转运体基因在气生根发育过程中显著上调表达,表明降解茎节处表皮细胞壁、利用硝酸盐在茎节处富集刺激根芽萌发是气生根生长发育调控机理之一。     

施氮水平对7S亚基缺失大豆根系形态和结瘤固氮的影响

为有效推广功能型大豆7S亚基缺失品种,以7S亚基缺失大豆品系东富2号为研究对象,设置4种施氮水平(纯N),N0(0 mg·kg~(-1))、N1(25 mg·kg~(-1))、N2(50 mg·kg~(-1))、N3(75 mg·kg~(-1)),采用桶栽法研究大豆根系形态和结瘤固氮对不同施氮水平的响应。结果表明:N1(25 mg·kg~(-1))水平下根系干重加大,根冠比增大,根瘤固氮潜力高,单株产量较高。N2(50 mg·kg~(-1))水平下根长、根表面积、根体积在生育后期增长较快,根系干重较大,根冠比低,固氮酶活性最高,单株籽粒产量最高。N3(75 mg·kg~(-1))水平下植株干重较大,无效生长较多,根瘤数少,固氮潜力和根冠比低,单株产量不高。综合籽粒产量和根系特性指标,功能型大豆7S亚基缺失品系东富2号的适宜施肥量为25~50 mg·kg~(-1)。     

Variation in TBX3 Gene Region in Dun Coat Color Polish Konik Horses

The Polish Konik is a primitive breed of horse, most likely descended directly from the wild extinct forest equus so-called Tarpan (Equus ferus ferus). The studbook of this breed is closed, and only individuals with blue dun in color with a well-developed black middorsal stripe and other primitive markings are entered. The aim of the presented study was the investigation of genetic background of dun phenotype in Polish Konik horses. A total of 93 Polish Koniks were investigated toward single nucleotide polymorphisms in a region of TBX3 gene by Sanger sequencing method. Obtained results confirmed that all investigated horses carry dun dilution genotype. Two new variants were identified which have not been previously described [WT/WT-G/G-A/G; WT/del-G/del-G/G]. Furthermore, our results showed in population of Polish Koniks occurrence of the rare genotype recently found in Estonian-native horses.     

间歇式双循环工厂化养殖系统构建及其养殖效果

为改善工厂化循环水养殖系统水质净化效果,提高养殖密度和成活率,构建了间歇式双循环工厂化养殖系统。通过间歇运行生物膜反应器增加水力停留时间,充分降解含氮污染物;连续运行弧形筛及时去除固体颗粒物。考察了该系统的启动过程及石斑鱼高密度养殖效果。启动初期,将硝化型生物絮团与海绵填料混合培养,生物膜22 d即可挂膜成功。以30.03 kg/m~3为初始养殖密度开展石斑鱼养殖试验,经66 d养殖,石斑鱼平均质量从(273.00±12.22)增至(552.52±107.04) g,最终养殖密度达到60.78 kg/m~3,成活率为100%。养殖过程中,生物膜逐渐适应养殖环境,氨氮、亚硝酸盐氮去除率从13.33%、14.84%增至93.73%、93.50%。此外,在弧形筛进水槽增加曝气形成曝气式弧形筛,可进一步除去细小颗粒物,有效控制养殖水体浊度。     

Effects of oral administration of Spirulina platensis and probiotics on serum immunity indexes,colonic immune factors,fecal odor,and fecal flora in mice

To evaluate the effects of Spirulina platensis and probiotics on growth, immunity indexes, fecal flora, and fecal odor in mice, 40 mice were randomly allotted to four groups, and each was administrated with nothing, S. platensis, probiotics, or both for 28 days, respectively. Then, many indexes were measured. The results showed that S. platensis was more effective (P < 0.001) than probiotics in improving mice's feed conversion ration (FCR). In immunity, probiotics administration increased (P < 0.042) serum IgE, IgM, IFN-γ, colonic AHR, TLR4, and NF-κB protein expression and decreased (P < 0.039) serum IL-1α, IL-21, IL-22, and colonic ARNT gene expression. However, the S. platensis showed weaker effect, which increased (P < 0.025) the serum IgE, IgM, TNF-α, and the colonic AHR and NF-κB protein expression, and decreased (P < 0.01) serum IL-21. Probiotics consumption decreased the fecal odor by decreasing (P < 0.02) fecal Escherichia coli, indole-3-acetic acid (IAA), and skatole contents, and the S. platensis decreased (P = 0.04) the IAA. These results indicated that oral administration of probiotics, S. platensis, or both of them in mice probably benefited body's immunity and reduced fecal odor. However, their mechanisms were still unclear and need further study.     

A genetic linkage map in southern‐by‐spring oat identifies multiple quantitative trait loci for adaptation and rust resistance

We developed 178 recombinant inbred lines from a southern‐by‐spring oat population designated as “TxH.” These lines were genotyped to generate a high‐quality linkage map that resolved 6,902 markers into 21 linkage groups that matched closely with the latest hexaploid oat consensus map. Three major quantitative trait loci (QTLs) affecting heading date were found in locations that are consistent with known QTLs and candidate genes, and two other QTLs affecting heading date were found in novel locations. Five QTLs affecting plant height were found. Both sets of QTLs are responsible for transgressive segregation observed for these two traits. Four QTLs affecting resistance to crown rust, caused by the pathogen Puccinia coronata f. sp. avenae, were identified. Two of these QTLs are consistent with known clusters of rust resistance genes, while two may represent new locations of novel rust resistance genes. A complete set of SNP sequences suitable for generating markers for molecular selection is provided.