印度木薯花叶病毒DNA在寄主植物中可能存在的形式

从印度木薯花叶病毒（ＩＣＭＶ）侵染的植物中纯化特异的核酸,经ＲＮＡａｓｅ,ＤＮＡａｓｃ,Ｎｕｃｌｅ－ａｓｅＳｌ,ＥｘｏｎｕｃｌｅａｓｅⅢ和ＥｃｏＲＩ酶切,Ｓｏｕｔｈｅｒｎ和Ｄｏｔｂｌｏｔｓ杂交证实,在感病的植株中,存在两种形式的病毒核酸：环状双链ＤＮＡ和环状单链ＤＮＡ,后者可能是病毒ＤＮＡ的（－）链,环状双链ＤＮＡ经限制性内切酶作用可得２．７ｋｂ的线性双链ＤＮＡ纯化的病毒核酸含ＤＮＡ１和ＤＮＡ２两个分子量相近的组份。     

鸭肝脂肪酸合成酶的NADPH底物抑制及作用动力学

己知动物脂肪酸合成酶的底物乙酰辅酶Ａ和丙二酰辅酶Ａ具有竞争性双底物抑制的乒乓机制。实验发现鸭肝脂肪酸合成酶的第三个底物ＮＡＤＰＨ也具有底物抑制,并研究了它的规律及与ＮＡＤＰＨ有关的稳态动力学。发现对于该酶的全反应,增加丙二酰辅酶Ａ浓度,降低环境盐浓度,均使ＮＡＤＰＨ底物抑制减少。但以ＮＡＤＰＨ作底物的酮酰还原和烯酰还原二步单独反应以及包含四步单独反应的乙酰乙酰辅酶Ａ还原反应都无ＮＡＤＰＨ底物抑制现象。ＮＡＤＰＨ底物抑制对丙二酰辅酶Ａ为竞争性,丙二酰辅酶Ａ底物抑制对ＮＡＤＰＨ为非竞争性。在全反应中ＮＡＤＰＨ和丙二酰辅酶Ａ之间发现为乒乓机制,在乙酰乙酰辅酶Ａ还原反应中,两个底物ＮＡＤＰＨ和乙酰乙酰辅酶Ａ之间则表现为序列反应机制。降低环境盐浓度使ＮＡＤＰＨ和丙二酰辅酶Ａ之间的乒乓机制向序列机制转化。在全反应中,ＮＡＤＰ产物抑制相对ＮＡＤＰ为竞争性,对丙二酰辅酶Ａ为非竞争性。     

Genomic concatemerization/deletion in rotaviruses: a new mechanism for generating rapid genetic change of potential epidemiological importance.

Three variants of group A rotavirus with large changes in their gene 5 structures have been analyzed at the molecular level. The first of these, P9 delta 5, was obtained during plaque purification undertaken as part of the biological cloning of a field isolate of virus. The gene 5 homolog in this isolate migrated just ahead of the normal segment 6 RNA, giving an estimated size of 1,300 bp. Molecular cloning and sequencing of this homolog revealed it to have a single 308-bp deletion in the center of the normal gene 5 sequence extending between nucleotides 460 and 768 of the normal gene sequence. This deletion caused a frameshift in the gene such that a stop codon was encountered 8 amino acids downstream of the deletion point, giving a predicted size for the protein product of this gene of 150 amino acids compared with the 490 amino acids of its normal-size counterpart. Attempts to detect this shortened protein in virus-infected cells were not successful, indicating that it was much less stable than the full-length protein and/or had suffered a large change in its antigenicity. The second two variants, brvA and brvE, were generated in an earlier study following the high-multiplicity passage of the UKtc strain of bovine rotavirus. Polyacrylamide gel electrophoresis analysis of these nondefective variants showed that brvA had a gene 5 homolog approximately equal in size to the normal RNA segment 2 (approximately 2,700 bp) and that brvE had a size of approximately 2,300 bp. Both variants showed changes in their gene 5 protein products, with brvA mimicking P9 delta 5 in failing to produce a detectable product whereas brvE produced a new virus-specific protein approximately 80 kDa in size. Full-length cDNA clones of the brvE gene 5 homolog were isolated, and analysis of their structure revealed a head-to-tail concatemerization of the normal gene 5 sequence with the first copy of the concatemer covering nucleotides 1 to 808 and the second covering nucleotides 92 to 1579, giving a total length of 2,296 bp. Sequencing across the junction region of the two copies of the gene showed that they were joined in frame to give a predicted combined open reading frame of 728 amino acids with the amino-terminal region consisting of amino acids 1 to 258 fused at the carboxy terminus to amino acids 21 to 490.(ABSTRACT TRUNCATED AT 400 WORDS)     

低剂量CO2激光对茄子,青椒叶绿体超微结构影响的初步观察

本试验用功率密度为825mw/cm~2的CO_2激光照射茄子干种子(含水量8.33％)13s、青椒干种子(含水量5.4％)3s,分别以Os为对照(ck)。照射后播于生长箱内,出苗10天分苗于玻璃温室内,于四叶期取样观察成熟叶片叶绿体的超微结构。发现激光照射处理的茄子、青椒的叶绿体的基质片层、基粒片层有明显的吸胀现象。     

用HPLC证实单糖在酸性处理中的变化

<正> 在碱性条件下,醛糖与酮糖之间会发生结构互变。然而美国学者Feather M S等经过十多年的研究,发现在酸性条件下,葡萄糖、甘露糖和果糖之间也有分子内的互变,并应用~3H放射标记等技术,提出了互变的反应机理,这一互变的存在给寡糖及多糖的组份分析带来了一个问题,即用酸性条件水解寡糖或多糖后,如何确定其中的葡萄糖、甘露糖或果糖     

TaTPP-7A positively feedback regulates grain filling and wheat grain yield through T6P-SnRK1 signalling pathway and sugar–ABA interaction

Grain size and filling are two key determinants of grain thousand-kernel weight (TKW) and crop yield, therefore they have undergone strong selection since cereal was domesticated. Genetic dissection of the two traits will improve yield potential in crops. A quantitative trait locus significantly associated with wheat grain TKW was detected on chromosome 7AS flanked by a simple sequence repeat marker of Wmc17 in Chinese wheat 262 mini-core collection by genome-wide association study. Combined with the bulked segregant RNA-sequencing (BSR-seq) analysis of an F₂ genetic segregation population with extremely different TKW traits, a candidate trehalose-6-phosphate phosphatase gene located at 135.0 Mb (CS V1.0), designated as TaTPP-7A, was identified. This gene was specifically expressed in developing grains and strongly influenced grain filling and size. Overexpression (OE) of TaTPP-7A in wheat enhanced grain TKW and wheat yield greatly. Detailed analysis revealed that OE of TaTPP-7A significantly increased the expression levels of starch synthesis- and senescence-related genes involved in abscisic acid (ABA) and ethylene pathways. Moreover, most of the sucrose metabolism and starch regulation-related genes were potentially regulated by SnRK1. In addition, TaTPP-7A is a crucial domestication- and breeding-targeted gene and it feedback regulates sucrose lysis, flux, and utilization in the grain endosperm mainly through the T6P-SnRK1 pathway and sugar–ABA interaction. Thus, we confirmed the T6P signalling pathway as the central regulatory system for sucrose allocation and source–sink interactions in wheat grains and propose that the trehalose pathway components have great potential to increase yields in cereal crops.     

Inhibitory role of bone marrow mesenchymal stem cells-derived exosome in non-small-cell lung cancer: microRNA-30b-5p,EZH2 and PI3K/AKT pathway

Exosomal microRNA (miRNA) exerts potential roles in non-small-cell lung cancer (NSCLC). The current study elucidated the role of miR-30b-5p shuttled by bone marrow mesenchymal stem cells (BMSCs)-derived exosomes in treating NSCLC. Bioinformatics analysis was performed with NSCLC-related miRNA microarray GSE169587 and mRNA data GSE74706 obtained for collection of the differentially expressed miRNAs and mRNAs. The relationship between miR-30b-5p and EZH2 was predicted and confirmed. Exosomes were isolated from BMSCs and identified. BMSCs-derived exosomes overexpressing miR-30b-5p were used to establish subcutaneous tumorigenesis models to study the effects of miR-30b-5p, EZH2 and PI3K/AKT signalling pathway on tumour growth. A total of 86 BMSC-exo-miRNAs were differentially expressed in NSCLC. Bioinfomatics analysis found that BMSC-exo-miR-30b-5p could regulate NSCLC progression by targeting EZH2, which was verified by in vitro cell experiments. Besides, the target genes of miR-30b-5p were enriched in PI3K/AKT signalling pathway. Animal experiments validated that BMSC-exo-miR-30b-5p promoted NSCLC cell apoptosis and prevented tumorigenesis in nude mice via EZH2/PI3K/AKT axis. Collectively, the inhibitory role of BMSC-derived exosomes-loaded miR-30b-5p in NSCLC was achieved through blocking the EZH2/PI3K/AKT axis.     

靶向RNA的CRISPR-Cas系统研究进展

CRISPR(clustered regularly interspaced short palindromic repeats)-Cas(CRISPR associated proteins)系统是细菌和古细菌抵抗噬菌体、质粒等外源遗传物质的一种适应性免疫系统,该系统利用一种特殊的RNA(CRISPR RNA,crRNA)指导的内切酶来切割与crRNA相互补的外源遗传物质,从而阻碍外源核酸的侵染。根据效应复合物组成形式的不同,CRISPR-Cas系统分为1类(Ⅰ型、Ⅳ型和Ⅲ型)和2类(Ⅱ型、Ⅴ型和Ⅵ型)两大类。目前已发现多个CRISPR-Cas系统具有非常强的特异靶向RNA编辑能力,如Ⅵ型CRISPR-Cas13系统和Ⅲ型CRISPR-Cas7-11系统。随着研究的深入,相关系统在RNA编辑领域应用日渐广泛,使其成为基因编辑的有力工具。本文介绍了靶向RNA的CRISPR-Cas系统的组成、结构、分子机制以及其潜在应用,这为更好地研究该类系统的作用机制奠定基础,也为后期开发为稳定的基因编辑工具提供新的思路。     

珍稀濒危植物蒜头果种子萌发特征与幼苗类型的研究

以去除果皮后阴干的珍稀濒危植物蒜头果种子为材料,在温室大棚沙池内进行层积处理,从层积处理开始至子叶出土的不同萌发阶段,考察蒜头果种子发育形态、贮藏物质积累、酶活性以及幼苗类型等变化特征,初步探讨其种子休眠形成原因。结果显示：(1)蒜头果种子从解除休眠开始至萌发形成幼苗的过程约需195 d,其中幼胚的形态发育后熟约需75 d,随后30 d内是种子集中萌发的时期,其发芽率达到最高(53.33%);依据种胚发育形态的标志特征将此过程划分为7个阶段(S1～S7阶段):S1阶段种子未萌发,S2阶段种胚“露白”,S3阶段胚根突破种皮长超过1 cm, S4阶段下胚轴与胚根连接处形成弯钩结构,S5阶段“S”型胚形成及胚根前端膨大,S6阶段种子不仅具有膨大的胚根且已有侧根的分化,S7阶段子叶脱落,胚芽出土,真叶出现。(2)蒜头果种子在湿沙层积过程中,种胚胚长和胚率从S1阶段的(5.49±1.57)mm和(19.48±5.72)%分别增加至S6阶段的(67.92±2.94)mm和(240.75±15.29)%,胚率平均增加了12.4倍,显示蒜头果种子的胚需要经历后熟过程才能萌发,属于胚后熟型种子。(3)从...