不同种植年限猕猴桃园土壤微生物功能多样性研究

陕西秦岭北麓是世界上最大的猕猴桃生产基地,长期相对单一的果树种植方式导致不同种植年限果园土壤微生态环境差异。研究猕猴桃园土壤微生物功能多样性随种植年限的变化特征,为果园土壤科学管理提供参考。采集不同种植年限猕猴桃根际土壤,应用平板菌落计数法和Biolog-Eco法研究土壤微生物的数量、种群以及功能多样性,并对土壤微生物功能多样性与土壤养分的相关性进行分析。结果显示,猕猴桃园土壤可培养微生物以细菌为主,其次是放线菌,真菌数量最少。随着种植年限的增加,细菌和放线菌数量显著降低,而真菌数量显著升高。微生物平均颜色变化率(average well-color development,AWCD)、微生物群落Shannon-Wiener指数(H′)、Simpson指数(D)及McIntosh指数(U)均随种植年限的增加而显著降低。主成分分析显示,不同种植年限猕猴桃园土壤微生物碳源利用特征明显不同,0～5 a、5～10 a和10～20 a的猕猴桃园土壤微生物群落分别被划分在载荷图的第一、第四和第三象限。猕猴桃园土壤微生物对糖类、氨基酸类和酯类的利用率相对较高,而对醇类、胺类和酸类的利用率相对较低。对第1主成分贡献大的碳源(|r|≥0.5)有11种,其中糖类占36%,氨基酸类和酯类均占18%。土壤微生物功能多样性与土壤养分相关性分析表明,土壤微生物功能多样性与土壤有机质正相关,与有效磷和速效钾负相关。结果表明,随种植年限的增加,猕猴桃园土壤微生物的数量和结构发生变化,微生物功能多样性降低,对碳源利用能力降低。鉴于土壤微生物功能多样性与土壤养分的相关性,应合理加大有机肥施用量,适量减少有效磷和速效钾的施用量。     

人嗅黏膜间充质干细胞来源外泌体的分离鉴定及生物学特性研究

外泌体是指释放到细胞外微环境中的直径约50～130 nm的纳米级的膜性囊泡。嗅黏膜间充质干细胞（olfactory mucosa mesenchymal stem cells,OM-MSCs）作为一类新发现的间充质干细胞,在许多疾病中均具有治疗作用,且其内在机制与其旁分泌的外泌体密切相关,但OM-MSCs外泌体的分离、鉴定及生物学特性的研究尚未见报道。本研究采用超速离心法提取OM-MSCs培养液中的外泌体,应用流式细胞术及免疫荧光进行细胞鉴定后,分别用透射电子显微镜、纳米粒径分析及Western印迹对外泌体形态、颗粒大小和表面的特异性分子标志进行分析鉴定。采用CCK8增殖实验,Western印迹和划痕实验,分析其对人脑微血管内皮细胞增殖和迁移的影响。电镜、Western 印迹和纳米粒径分析的结果显示：OM-MSCs来源外泌体形态多为圆形,直径约为40～150 nm;表达外泌体标记物CD63,CD81;CCK-8法检测显示：不同浓度的OM-MSCs源外泌体可提高人脑微血管内皮细胞的增殖活性,且其增殖促进作用具有浓度依赖性（P<0.05）。Western 印迹检测结果显示：相比空白对照组,OM-MSCs源外泌体可显著提高内皮细胞的增殖细胞核抗原蛋白质水平表达（P<0.01）,细胞划痕实验结果显示,OM-MSCs源外泌体可增强内皮细胞的迁移能力,且高于对照组（P<0.01）。本研究表明：通过超速离心法可以分离纯化获得OM-MSCs源外泌体,且该外泌体具有促进人脑微血管内皮细胞迁移和增殖的作用。     

Probiotic Clostridium butyricum Improves the Growth Performance,Immune Function,and Gut Microbiota of Weaning Rex Rabbits

Probiotics could promote animal growth and enhance immune function. This study investigated the effects of Clostridium butyricum (CB) on the growth performance, intestinal immune, and gut microbiota of weaning rex rabbits. A total of 60 healthy female rabbits (5-month-old) were divided equally into four groups and mated on the same day: control group (CTRL, fed with basal feed), low-dose group (LDG, fed with basal feed + 1.0 × 10³ CFU/g CB), middle-dose group (MDG, fed with basal feed + 1.0 × 10⁴ CFU/g CB), and high-dose group (HDG, fed with basal feed + 1.0 × 10⁵ CFU/g CB). Then, 30 weaning rex rabbits (35-day-old) were collected from each group for this experiment, and they were offered the same feeds as their mother. The results demonstrated that high-dose CB treatment significantly increased average daily weight gain of weaning rex rabbits. Further studies suggested that CB enhanced small intestinal digestive enzyme activity and improved mucosal morphology and antioxidant status. Supplemented with CB, small intestinal barrier function was maintained with the upregulation of mRNA levels of ZO-1, claudin, and occludin as well as the increase of sIgA production. Moreover, the relative expressions of MyD88, TLR2, and TLR4 were elevated in HDG; simultaneously, pro-inflammatory cytokines including IL-6, INF-γ, and TNF-α were decreased after CB administration. In addition, CB showed beneficial effects in improving weaning rex rabbit intestinal microflora via increasing the abundance of beneficial bacteria. Therefore, our results indicated CB can promote rex rabbit growth, which is likely to the enhancement of immune function and the improvement of intestinal microbiota.

     

Maternal genomic variability of the wild boar (Sus scrofa) reveals the uniqueness of East‐Caucasian and Central Italian populations

The phylogeography of the European wild boar was mainly determined by postglacial recolonization patterns from Mediterranean refugia after the last ice age. Here we present the first analysis of SNP polymorphism within the complete mtDNA genome of West Russian (n = 8), European (n = 64), and North African (n = 5) wild boar. Our analyses provided evidence of unique lineages in the East‐Caucasian (Dagestan) region and in Central Italy. A phylogenetic analysis revealed that these lineages are basal to the other European mtDNA sequences. We also show close connection between the Western Siberian and Eastern European populations. Also, the North African samples were clustered with the Iberian population. Phylogenetic trees and migration modeling revealed a high proximity of Dagestan sequences to those of Central Italy and suggested possible gene flow between Western Asia and Southern Europe which was not directly related to Northern and Central European lineages. Our results support the presence of old maternal lineages in two Southern glacial refugia (i.e., Caucasus and the Italian peninsula), as a legacy of an ancient wave of colonization of Southern Europe from an Eastern origin.     

Development of high‐resolution DNA barcodes for Dioscorea species discrimination and phylogenetic analysis

The genus Dioscorea is widely distributed in tropical and subtropical regions, and is economically important in terms of food supply and pharmaceutical applications. However, DNA barcodes are relatively unsuccessful in discriminating between Dioscorea species, with the highest discrimination rate (23.26%) derived from matK sequences. In this study, we compared genic and intergenic regions of three Dioscorea chloroplast genomes and found that the density of SNPs and indels in intergenic sites was about twice and seven times higher than that of SNPs and indels in the genic regions, respectively. A total of 52 primer pairs covering highly variable regions were designed and seven pairs of primers had 80%–100% PCR success rate. PCR amplicons of 73 Dioscorea individuals and assembled sequences of 47 Dioscorea SRAs were used for estimating intraspecific and interspecific divergence for the seven loci: The rpoB‐trnC locus had the highest interspecific divergence. Automatic barcoding gap discovery (ABGD), Poisson tree processes (PTP), and generalized mixed Yule coalescence (GMYC) analysis were applied for species delimitation based on the seven loci and successfully identified the majority of species, except for species in the Enantiophyllum section. Phylogenetic analysis of 51 Dioscorea individuals (28 species) showed that most individuals belonging to the same species tended to cluster in the same group. Our results suggest that the variable loci derived from comparative analysis of plastid genome sequences could be good DNA barcode candidates for taxonomic analysis and species delimitation.     

Microsatellite‐based analysis of genetic structure and gene flow of Mythimna separata (Walker) (Lepidoptera: Noctuidae) in China

The oriental armyworm, Mythimna separata, is a serious agricultural pest in China. Seasonal and roundtrip migration has recently led to sudden, localized outbreaks and crop losses. To evaluate genetic differentiation between populations in eastern and western China and elucidate gene flow, the genetic structure of 20 natural populations from nine provinces was examined using seven microsatellite markers. The results indicated high genetic diversity. However, little to moderate (0 < F_ST < 0.15) genetic differentiation was detected, and there was no correlation between genetic distance and geographical distance. Bayesian clustering analysis identified three groups whereas discriminant analysis of principal components identified ten clusters that were considered as two clear‐cut clusters and one admixed group. Gene flow occurred frequently in most population pairs, and an asymmetrical migration rate was detected in several pairwise population comparisons. The bottleneck test showed that few populations had experienced recent bottlenecks. Correspondingly, large‐scale and long‐distance migration of M. separata has caused low genetic differentiation and frequent gene exchange. Our findings are important for studying genetic evolution and help to improve predictions of M. separata outbreaks in China.     

脐带血干细胞移植治疗非恶性血液病进展

异基因造血干细胞移植（allo-HSCT）是治愈多种非恶性病的有效方法。脐带血干细胞（UCB）具有免疫原性低、人类白细胞抗原不合耐受性好、移植物抗宿主反应发生率低以及获取相对快捷等特点,可作为非恶性血液疾病患者allo-HSCT的来源。本文简要综述脐血干细胞移植在原发性免疫缺陷病、遗传性骨髓衰竭、遗传代谢病以及自身免疫性疾病等非恶性血液疾病的治疗效果。     

Nemo-like Kinase Drives Foxp3 Stability and Is Critical for Maintenance of Immune Tolerance by Regulatory T Cells

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海南发现新入侵害虫——橘绵粉虱Aleurothrixus floccosus(Maskell)

橘绵粉虱是一种危险性的入侵生物,已于2018年11月在我国海南文昌的番石榴上发现。本文记述了橘绵粉虱各虫态的形态特征、野外鉴别特征,并列出了其分布及重要寄主植物,附有彩色生态照片。由于橘绵粉虱是柑橘类的重要害虫,具有向中国大陆扩散的可能性,我国南方柑橘产区应密切关注此虫动态。