AST同工酶的琼脂糖凝胶电泳NBT显色法

分布于细胞内线粒体及细胞质中的天门冬氨酸氨基转移酶(AST·EC·2·6·1·1)是人血清中含有的两种同工酶,分别称为AST-m和AST-c同工酶.AST-c电泳迁移率介于血清α-球蛋白与β-球蛋白之间.AST-m电泳迁移率相似于γ-球蛋白,琼脂糖凝胶电泳固蓝B染色法对m-AST检出率较低.用NBT显色法则可得到较好效果.     

Genomic insights into selection for heterozygous alleles and woody traits in Populus tomentosa

Heterozygous alleles are widespread in outcrossing and clonally propagated woody plants. The variation in heterozygosity that underlies population adaptive evolution and phenotypic variation, however, remains largely unknown. Here, we describe a de novo chromosome-level genome assembly of Populus tomentosa, an economic and ecologically important native tree in northern China. By resequencing 302 natural accessions, we determined that the South subpopulation (Pop_S) encompasses the ancestral strains of P. tomentosa, while the Northwest subpopulation (Pop_NW) and Northeast subpopulation (Pop_NE) experienced different selection pressures during population evolution, resulting in significant population differentiation and a decrease in the extent of heterozygosity. Analysis of heterozygous selective sweep regions (HSSR) suggested that selection for lower heterozygosity contributed to the local adaptation of P. tomentosa by dwindling gene expression and genetic load in the Pop_NW and Pop_NE subpopulations. Genome-wide association studies (GWAS) revealed that 88 single nucleotide polymorphisms (SNPs) within 63 genes are associated with nine wood composition traits. Among them, the selection for the homozygous AA allele in PtoARF8 is associated with reductions in cellulose and hemicellulose contents by attenuating PtoARF8 expression, and the increase in lignin content is attributable to the selection for decreases in exon heterozygosity in PtoLOX3 during adaptive evolution of natural populations. This study provides novel insights into allelic variations in heterozygosity associated with adaptive evolution of P. tomentosa in response to the local environment and identifies a series of key genes for wood component traits, thereby facilitating genomic-based breeding of important traits in perennial woody plants.     

巨噬细胞激活及钙作用的初步研究

经TG诱发的小鼠腹腔渗出液的巨噬细胞及人的巨噬细胞样细胞株U937受PAF(100ng ml).Zymosan A(0.25mg ml).Can A(50μg ml)LPS(1μg ml)等作用后.能引起胞内游离Ca~(2-)浓度的增加,巨噬细胞内酸性磷酸酶增多,细胞骨架更为舒展、丰满.胞内游离Ca~(2-)的增加是由于胞内钙库的释放与胞外钙的内流.上述困子作用后,可使巨噬细胞产生呼吸爆发.其中.Zymosan A的作用尤为强烈.同时还出现胞膜流动性的降低、当胞外环境中有Ca~(2-)时.可增强巨噬细胞的呼吸爆发.以上提示:在巨噬细胞的激活中Ca~(2-)具有重要的作用.     

秦艽体细胞胚不同发育阶段几种同工酶的特性

以秦艽（ＧｅｎｔｉａｎａｃｒａｓｓｉｃａｕｌｉｓＤｕｔｈｉｅｅｘＢｕｒｋｉｌｌ）无菌苗幼嫩茎叶为外植体,在ＭＳ培养基上诱导出胚性愈伤组织及体细胞胚。本文比较了秦荒体细胞胚发育过程中几种酶的活性及同工酶变化。过氧化物酶与细胞色素氧化酶均在球形胚阶段出现一活性高峰;酯酶同工酶在体细胞胚发育过程中伴有特征带Ｅ＿２、Ｅ＿６的出现;酸性磷酸酯酶的活性与体细胞胚发育程度呈正相关。所以认为只要综合运用这几种同工酶的实验数据,就可以灵敏检测体细胞胚的发育进程。     

QTL identification of grain protein concentration and its genetic correlation with starch concentration and grain weight using two populations in maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.)

Protein is one of the three main storage chemical components in maize grains, and is negatively correlated with starch concentration (SC). Our objective was to analyse the influence of genetic backgrounds on QTL detection for protein concentration (PC) and to reveal the molecular genetic associations between PC and both SC and grain weight (GWP). Two hundred and eighty-four (Pop1) and 265 (Pop2) F_2:3 families were developed from two crosses between one high-oil maize inbred GY220 and two normal maize inbreds 8984 and 8622 respectively, and were genotyped with 185 and 173 pairs of SSR markers. PC, SC and GWP were evaluated under two environments. Composite interval mapping (CIM) and multiple interval mapping (MIM) methods were used to detect single-trait QTL for PC, and multiple-trait QTL for PC with both SC and GWP. No common QTL were shared between the two populations for their four and one PC QTL. Common QTL with opposite signs of effects for PC and SC/GWP were detected on three marker intervals at bins 6.07–6.08, 8.03 and 8.03–8.04. Multiple-traits QTL mapping showed that tightly-linked QTL, pleiotropic QTL and QTL having effects with opposite directions for PC and SC/GWP were all observed in Pop1, while all QTL reflected opposite effects in Pop2.     

Epidermal restriction confers robustness to organ shapes

The shape of comparable tissues and organs is consistent among individuals of a given species, but how this consistency or robustness is achieved remains an open question. The interaction between morphogenetic factors determines organ formation and subsequent shaping, which is ultimately a mechanical process. Using a computational approach, we show that the epidermal layer is essential for the robustness of organ geometry control. Specifically, proper epidermal restriction allows organ asymmetry maintenance, and the tensile epidermal layer is sufficient to suppress local variability in growth, leading to shape robustness. The model explains the enhanced organ shape variations in epidermal mutant plants. In addition, differences in the patterns of epidermal restriction may underlie the initial establishment of organ asymmetry. Our results show that epidermal restriction can answer the longstanding question of how cellular growth noise is averaged to produce precise organ shapes, and the findings also shed light on organ asymmetry establishment.     

TGFβ2 is a prognostic‐related biomarker and correlated with immune infiltrates in gastric cancer

TGFβ2 is an essential regulator of immune cell functionality, but the mechanisms whereby it drives immune infiltration in gastric cancer remain uncertain. The Oncomine and Tumor Immunoassay Resource (TIMER) databases were used for assessing the expression of TGFβ2, after which TIMER was used to explore the relationship between TGFβ2 and tumour immune infiltration. Finally, we assessed how TGFβ2 expression correlated with the expression of a set of marker genes associated with immune infiltration using TIMER and GEPIA. We determined TGFβ2 expression to be significantly correlated with outcome in multiple types of cancer in the Cancer Genome Atlas (TCGA), with the effect being particularly pronounced in gastric cancer. Furthermore, elevated TGFβ2 expression was found to be significantly correlated with gastric cancer N staging, and with the expression of a variety of immune markers associated with particular immune cell subsets. These results indicate that TGFΒ2 is associated with patient outcome and tumour immune cell infiltration in multiple cancer types. This suggests that TGFβ2 is a key factor which governs immune cell recruitment to gastric cancer tumours, potentially playing a vital role in governing immune cell infiltration and thus representing a valuable prognostic biomarker in gastric cancer patients.     

染色质免疫共沉淀实验方法

染色质免疫共沉淀(ChIP)技术是一种检测蛋白质与DNA结合的实验技术。该方法可以先进行样品交联, 然后将蛋白质与DNA复合物进行随机DNA切断, 再借助免疫学方法特异性富集与目的蛋白相结合的DNA片段, 从而检测转录因子等目的蛋白质与DNA的结合情况, 鉴定基因启动子或其它DNA结合位点。该方法同时也可应用于研究基因组特定位点的组蛋白修饰情况。该文介绍了依赖交联固定的常规免疫共沉淀(X-ChIP), 以及适用于10³细胞级别微量实验材料的基于微球菌核酸酶非交联免疫共沉淀(ULI-NChIP)具体操作过程和注意事项。     

A Phytophthora capsici effector suppresses plant immunity via interaction with EDS1

EDS1 (Enhanced Disease Susceptibility 1) plays a crucial role in both effector-triggered immunity activation and plant basal defence. However, whether pathogen effectors can target EDS1 or an EDS1-related pathway to manipulate immunity is rarely reported. In this study, we identified a Phytophthora capsici Avirulence Homolog (Avh) RxLR (Arg-any amino acid-Leu-Arg) effector PcAvh103 that interacts with EDS1. We demonstrated that PcAvh103 can facilitate P. capsici infection and is required for pathogen virulence. Furthermore, genetic evidence showed that PcAvh103 contributes to virulence through targeting EDS1. Finally, PcAvh103 specifically interacts with the lipase domain of EDS1 and can promote the disassociation of EDS1–PAD4 (Phytoalexin Deficient 4) complex in planta. Together, our results revealed that the P. capsici RxLR effector PcAvh103 targets host EDS1 to suppress plant immunity, probably through disrupting the EDS1–PAD4 immune signalling pathway.     

Two Phytophthora parasitica cysteine protease genes,PpCys44 and PpCys45, trigger cell death in various Nicotiana spp. and act as virulence factors

Proteases secreted by pathogens have been shown to be important virulence factors modifying plant immunity, and cysteine proteases have been demonstrated to participate in different pathosystems. However, the virulence functions of the cysteine proteases secreted by Phytophthora parasitica are poorly understood. Using a publicly available genome database, we identified 80 cysteine proteases in P. parasitica, 21 of which were shown to be secreted. Most of the secreted cysteine proteases are conserved among different P. parasitica strains and are induced during infection. The secreted cysteine protease proteins PpCys44/45 (proteases with identical protein sequences) and PpCys69 triggered cell death on the leaves of different Nicotiana spp. A truncated mutant of PpCys44/45 lacking a signal peptide failed to trigger cell death, suggesting that PpCys44/45 functions in the apoplastic space. Analysis of three catalytic site mutants showed that the enzyme activity of PpCys44/45 is required for its ability to trigger cell death. A virus-induced gene silencing assay showed that PpCys44/45 does not induce cell death on NPK1 (Nicotiana Protein Kinase 1)-silenced Nicotiana benthamiana plants, indicating that the cell death phenotype triggered by PpCys44/45 is dependent on NPK1. PpCys44- and PpCys45-deficient double mutants showed decreased virulence, suggesting that PpCys44 and PpCys45 positively promote pathogen virulence during infection. PpCys44 and PpCys45 are important virulence factors of P. parasitica and trigger NPK1-dependent cell death in various Nicotiana spp.