Analysis of RNA-Dependent RNA Polymerase Sequence of Infectious Flacherie Virus Isolated in China and Its Expression in BmN Cells

Full gene sequence of RNA-dependent RNA polymerase （RdRp） from Bombyx mori infectious flacherie virus isolated in Zhejiang Province, China （Zhejiang01/CHN/2002） was cloned. The sequence was 1 920 nucleotides in length coding 639 amino acid residues. Sequences comparison of RdRp showed Zhejiang01/CHN/2002 was 99.7% nucleotide sequence and 99.1% amino acids sequence homology with Japanese strain. The RdRp sequence was aligned with 8 representative picorna（-like） viruses and 8 highly conserved regions were detected. The result indicated their relevance function. Phylogenetic tree of 14 picorna（-like） viruses which RdRp presumed protein sequences revealed that the viruses from Iflavirus genus formed an independent clade. The RdRp was successfully expressed in BmN cells using Bac-to-Bac expression system.     

夏枯草不同器官主要药用成分积累规律

为揭示夏枯草不同器官主要药用成分的积累规律,采用紫外-可见分光光度法测定夏枯草不同器官不同时期总黄酮、熊果酸、齐墩果酸、迷迭香酸和咖啡酸含量。结果表明,总黄酮、熊果酸、齐墩果酸、迷迭香酸和咖啡酸在夏枯草根、茎、叶和花中均有分布,但含量不同;总黄酮在叶中含量最高,齐墩果酸在根中含量最高;熊果酸、迷迭香酸和咖啡酸在花中含量最高。总黄酮、熊果酸、齐墩果酸、迷迭香酸和咖啡酸在同一器官不同生长时期含量不同。研究认为夏枯草根、茎、叶、花中都分布有药用成分,可全草入药,并且在终花期至果实成熟期各部位药用成分含量较高,可在此期间采收。     

TBTV卫星RNA不同分离物的序列测定及系统进化分析*

  利用RT-PCR方法对烟草丛顶病毒（tobacco bushy top virus,TBTV）卫星RNA不同分离物进行扩增和克隆测序,同时运用DNAMAN,MEGA,ClustalX等生物软件对TBTV和其它幽影病毒的卫星RNA核苷酸序列进行同源性比较,构建了幽影病毒属成员卫星RNA的系统进化树。E结果显示：TBTV不同分离物卫星RNA的核苷酸同源性为792%～990%。它们与幽影病毒属其它成员卫星RNA的核苷酸同源性为325%～441%。通过构建幽影病毒属成员卫星RNA的系统进化树可以看出：TBTV的卫星RNA与GRV的卫星RNA亲缘关系最近,与CMoMV的亲缘关系最远。     

分光光度法测定凤仙花中总黄酮的含量

［目的］建立能够准确测定凤仙花中总黄酮含量的方法。［方法］分别以芦丁、山柰酚为对照品,可见分光光度法是采用AlCl3显色,在412 nm波长处测定总黄酮的含量;紫外分光光度法在270 nm处测定总黄酮的含量。［结果］以芦丁、山柰酚为对照品采用可见分光光度法测得总黄酮含量的平均回收率分别为98.52%、99.72%,RSD分别为2.38%、3.14%;紫外分光光度法测得总黄酮含量的平均回收率分别为97.53%、98.75%,RSD分别为2.71%、2.86%。［结论］以山柰酚为对照品、采用AlCl3为显色剂的可见分光光度法是一种较理想的测定凤仙花中总黄酮含量的方法。     

非编码RNA检测技术的研究进展

非编码RNA（non-coding RNA,ncRNA）广泛存在于生物体中,不编码蛋白质,以RNA的形式在许多生命过程中发挥着重要的作用。随着越来越多ncRNA生物学功能的揭示,检测ncRNA在不同生理和病理状态下表达谱的变化,已迅速成为研究的热点。为了将ncRNA更好地应用于农业生产和生物医学研究,笔者对ncRNA最新检测技术的原理与应用进行了综述。     

Mutations in FgPrp6 suppressive to the Fgprp4 mutant in Fusarium graminearum

The pre-mRNA processing factor Prp6 is an essential component of the U4/U6.U5 tri-small nuclear ribonucleoprotein(snRNP).In a previous study,mutations were identified in the PRP6 ortholog in four suppressors of Fgprp4 that was deleted of the only kinase FgPrp4 among the spliceosome components in the plant pathogenic fungus Fusarium graminearum.In this study,we identified additional suppressor mutations in FgPrp6 and determined the suppressive effects of selected mutations.In total,12 mutations o...     

RNAi技术在转基因西瓜抗病毒病上的研究与应用

对RNAi技术的概念、研究进展及西瓜病毒病的危害及其发生特点、RNAi技术在西瓜抗病毒病上的研究进展进行了阐述,并对存在的一些问题作了简要探讨与展望。     

Effects of Smo gene silencing on cell activity and apoptosis of human cervical carcinoma HeLa cells

AIM: To investigate the effect of Hedgehog (Hh) signaling pathway on the viability and apoptosis of cervical carcinoma cells by shRNA technique to knock down Smoothened (Smo) gene. METHODS: Smo shRNA was used to transfect the cervical carcinoma HeLa cells. The expression of Smo and Gli1 at mRNA and protein levels in the HeLa cells was determined by RT-PCR and Western blot, respectively. The effect of Smo gene silencing on the growth of the cells was measured by MTT assay. The apoptosis and cell cycle were determined by flow cytometry. RESULTS: Compared with control group, the mRNA and protein expression of Smo and Gli1 were evenly reduced obviously after transfected with Smo shRNA for 72 h (P<0.05). The viability of HeLa cells transfected with Smo shRNA was significantly inhibited. The percentages of the cells in G₀/G₁ phase and early apoptosis rate were obviously higher in Smo shRNA transfection group than those in control group. CONCLUSION: Smo gene silencing effectively inhibits the cell growth and induces the apoptosis of human cervical carcinoma cells.     

适用于转录组测序的大白菜小孢子总RNA提取方法筛选

为了找到适用于转录组测序要求的大白菜小孢子总RNA 提取方法，以出胚率较高的吉红82 为试材，以热激诱导处理后的小孢子为研究对象，比较分析了TRIzol 法、改良CTAB 法及超纯试剂盒法3 种方法的RNA 提取效果。结果表明：TRIzol法（磨样）和改良CTAB 法提取的RNA 浓度较高，但完整性较差；超纯RNA 提取试剂盒法提取的RNA 质量最高，且完整性好，能够满足转录组测序的要求。     

基于紫花苜蓿混播草地的全草型肉羊放牧育肥模式案例研究

本研究旨在探讨紫花苜蓿（Medicago sativa L.）混播草地放牧利用的生产潜力和集成全草型集约化肉羊放牧育肥技术模式。2017-2018年，在位于河北省廊坊市的中国农业科学院国际农业高新技术产业园内开展了2年羔羊育肥的划区轮牧试验，研究了紫花苜蓿混播草地的地上生物量、育肥羊增重以及羊肉品质的变化规律。结果表明：混播草地地上生物量超过11.2 tDM·hm^-2，5月份紫花苜蓿混播草地的粗蛋白为20.92%，每公顷紫花苜蓿混播草地可满足51只育肥羊、放牧育肥150天，每只增重30 kg所需的干物质、代谢能和粗蛋白三个方面的全部需求。与此同时，紫花苜蓿混播草地还可以显著增加羊肉中欧米伽3型多不饱和脂肪酸的含量，n-6/n-3比例为2.72，羊肉健康品质显著提高。以紫花苜蓿混播草地为基础的全草型肉羊放牧育肥技术具有广阔的应用前景，对我国苜蓿产业以及草地畜牧业发展具有重要意义。