小麦TaeEF1β基因的克隆、同源性及表达分析

真核翻译延伸因子(eukaryotic translation elongation factor,eEFs)是一种重要的多功能调控蛋白,eEF1β是eEF1的组成部分,在蛋白质生物合成过程中发挥着重要的作用。本文通过RT-PCR扩增克隆小麦(Triticum aestivum L.)的eEF1β基因,并命名为TaeEF1β。氨基酸同源性分析发现,TaeEF1β具有高度保守性,且其保守结构域位于137~226 aa处。qRT-PCR结果表明,中国小麦花叶病毒(Chinese wheat mosaic virus,CWMV)侵染小麦植株后,可以诱导TaeEF1β基因转录水平的上调表达。另外,本文也进一步分析了TaeEF1β基因在小麦根、茎、叶的表达水平和CWMV侵染不同时间点的表达情况。     

朝鲜黄杨种源试验苗期效果初报

利用引进的锦州、沈阳、大连、鞍山4个产地朝鲜黄杨(Buxus microphylla var.koreana)种源进行播种试验,对幼苗出苗率、保存率、苗高、地径、分枝数、冠幅调查测量。结果表明:4个种源的出苗率、当年苗高、分枝数、冠幅均存在显著差异,地径差异不显著。沈阳种源播种出苗率最高,为75.48%;沈阳、大连种源当年苗高14.53 cm、14.08 cm,分枝数9.43条、9.21条,冠幅12.02 cm、10.00 cm,显著高于鞍山、锦州种源;苗木越冬保存率均在90%以上。沈阳种源表现最好。鞍山和锦州种源表现较差。     

谷氨酰胺对斜带石斑鱼GF-1细胞中C-Myc蛋白的表达与神经坏死病毒复制的影响

为探讨C-Myc表达、谷氨酰胺代谢和神经坏死病毒复制三者之间的关系,本研究首先克隆了斜带石斑鱼鳍条细胞(GF-1)中的C-Myc基因(GF-1-C-Myc),结果显示GF-1-CMyc基因cDNA全长814 bp,开放阅读框(ORF)为285 bp,编码95个氨基酸(aa),有亮氨酸拉链结构域与螺旋-环-螺旋(HLH)结构域。实验表达和纯化了GF-1-C-Myc蛋白,并制备其多克隆抗体。采用实时定量PCR技术(qRT-PCR)与免疫印迹法(WB)检测了GF-1-C-Myc基因的表达和神经坏死病毒的复制。结果显示,缺乏谷氨酰胺会同时抑制GF-1-C-Myc基因的表达和神经坏死病毒(NNV)的复制,添加谷氨酰胺可同时促进GF-1-C-Myc的表达和NNV的复制;此外,NNV感染可上调GF-1-C-Myc基因的表达,并显著消耗GF-1细胞培养液中的谷氨酰胺。研究表明,GF-1-C-Myc基因可调控宿主谷氨酰胺代谢,从而有利于神经坏死病毒的复制。本结果为防控NNV的感染提供了参考。     

Crude garlic extract significantly inhibits replication of grapevine viruses

Efforts to control viral diseases of grapevine include the production of certified material and development of virus-resistant transgenic grapevines. However, effective antiviral agents, once the viruses have infected the plants, are still lacking. This study shows that a crude garlic extract has significant antiviral activity against grapevine viruses. Replication of grapevine leafroll-associated virus 2 (GLRaV-2) was obviously inhibited in grapevine cv. Cabernet Sauvignon calli treated with diluted (1:100) garlic extract. The relative RNA levels of GLRaV-2 and grapevine fleck virus (GFkV) in cv. Summer Black grapevine in in vitro-grown plantlets 10 days after treatment with diluted (1:100) garlic extract were about 22% and 20%, respectively, of that in controls. The viral RNA accumulation of GLRaV-2, GFkV, grapevine virus A (GVA), grapevine fanleaf virus (GFLV) and grapevine rupestris stem pitting-associated virus (GRSPaV) in field-grown grapevine cv. Centennial Seedless plants sprayed with diluted (1:100) garlic extract were about 31–40%, 26–38%, 18–31%, 17–42% and 15–18%, respectively, of that in controls. Moreover, the garlic extract treatment led to a significant decrease in viral RNA accumulation of GLRaV-3, GLRaV-2, GVA, GFkV, GFLV, GRSPaV and grapevine Pinot Gris virus in pot-grown grapevine cv. Shine Muscat plants, and viral disease symptoms in these plants were obviously attenuated. In addition, this extract significantly induced expression of pathogenesis-related protein genes and stimulated activity of antioxidant enzymes in grapevines. Taken together, these results indicate that the crude garlic extract acts as a significant inhibitor against a broad range of grapevine viruses.     

基于纳米碳添加的减量施肥模式对寒地水稻产量及氮肥效率的影响

为解决减量施肥造成北方寒地水稻肥料供给不足和产量低的问题,以纳米碳增效剂为材料,采用仪器快速检测和植株常规测定分析方法,研究纳米碳添加的常规施肥和减量施肥模式对水稻产量性状和肥料利用效率的影响。结果表明：与常规施肥(CK)相比,纳米碳添加的常规施肥模式(SM-A)、减施氮肥10%(SM-B)和减施氮肥20%(SM-C)对水稻植株形态特征、光合生理特性、产量性状和肥料利用效率产生显著的影响,其中以SM-B增幅最大,SM-A次之,SM-C最小;SM-A、SM-B和SM-C增加经济效益分别为563.56、1374.20和-1391.08元/hm ²,以SM-B增效值最高。本研究表明,SM-B能改善生育期光合生理特性,提高植株产量和肥料利用效率,是寒地水稻较为理想的减量施肥安全利用模式。     

Virucidal effects of various agents—including protease—against koi herpesvirus and viral haemorrhagic septicaemia virus

In a search for alternative, environmentally friendly and effective disinfecting agents, a commercially available protease—Neutrase^®—was tested in this work for inactivation of koi herpesvirus (KHV) and of viral haemorrhagic septicaemia virus (VHSV). For comparison, the stability of these viral pathogens in similar configurations at various pH values and concentrations of peracetic acid or quicklime, typically used for disinfection, was tested. Therefore, virus suspensions were incubated with various concentrations of different agents for 24 hr and the titre of the remaining infectious particles was determined by virus titration. Furthermore, the treatment of both viruses, with the agents at concentrations that were previously appointed as effective, was also examined in the presence of solid material (quartz sand). All procedures investigated in this study, including the protease treatment, were able to reduce the titre of KHV and VHSV below the detection limit of the titration. Although further studies are necessary, this is the first report of the application of a protease for the inactivation of the selected fish pathogens, demonstrating the great potential of the latter for disinfection.     

Molecular detection of sugarcane striate virus and sugarcane white streak virus and their prevalence in the Miami World Collection of sugarcane and related grasses

Viruses in the genus Mastrevirus (family Geminiviridae), including those infecting sugarcane, have natural geographical ranges almost exclusively restricted to Africa and the Indian Ocean islands off the African coast. Only sugarcane white streak virus (SWSV) in Barbados and sugarcane striate virus (SStrV) in Florida and Guadeloupe are known to infect a few sugarcane varieties in the Western Hemisphere. In this study, PCR assays were developed to detect these two viruses in sugarcane. Five hundred and seventy-one DNA samples from Saccharum species and interspecific hybrids from the Miami World Collection of sugarcane and related grasses were tested for the presence of SStrV and SWSV by PCR. No variety was found infected by SWSV but SStrV was detected in 19 varieties. PCR data were confirmed by sequencing amplified fragments (248 bp). These fragments shared 93%–100% nucleotide identity with SStrV sequences from the GenBank database. SStrV isolates were distributed in six phylogenetic groups, including the four strains of the virus. Most varieties infected by SStrV originated from Asia, thus confirming a previous hypothesis stating that this virus originated from this continent. Absence of SStrV in commercial sugarcane in Florida also suggested that this virus has not been spread in this location, while infected plants have been present for several decades.     

快速灵敏的甘蔗线条花叶病毒免疫学检测体系的建立

 甘蔗花叶病广泛存在于我国甘蔗种植区,严重影响甘蔗产业的高质量发展。近年来甘蔗线条花叶病毒在蔗区肆虐,尽管针对其的血清学检测技术已经建立,但是快速、准确、高通量的检测方法亟待发掘。本研究制备了^SCSMVCP的抗血清,特异性高,与引起甘蔗花叶病的另两种病原 (高粱花叶病毒和甘蔗花叶病毒) 间没有血清学交叉反应。基于该多克隆抗体,建立了直接抗原包被的ELISA、斑点杂交、Western blot和基于多抗的免疫试纸条检测技术。开发的免疫试纸条检测技术能快速、准确、高通量应用于田间病毒鉴定。本文提供了基于血清学的快速、准确、高通量,且便捷的甘蔗线条花叶病毒检测技术,有助于我国蔗区甘蔗花叶病的监测与防控。     

建兰花叶病毒和齿兰环斑病毒的检测技术及脱毒方法研究进展

为了弄清目前建兰花叶病毒(CymMV)和齿兰环斑病毒(ORSV)病毒检测和脱毒的研究概况,本研究详细阐述了2种病毒的检测方法、采用的脱毒技术和脱毒效率,分析比较了不同技术存在的弊端,指出现有的脱毒方法已不能满足兰科植物脱毒研究的需要。在此基础上,进一步提出应采用安全、可靠的新型脱毒技术来解决目前兰科植物产业瓶颈问题。超低温脱毒技术是近年来发展迅速的脱毒技术,具有独特优势,目前已在多种植物上成功应用,该方法的应用有望解决2种病毒脱毒方面存在的问题。     

十里香茶离体培养及再生体系研究

为了保护珍贵的十里香茶树资源,掌握利于十里香茶的最优组培条件,本研究对十里香茶组培过程中外植体类型、消毒时间、培养基添加物及培养条件进行了筛选。结果表明,采用轻微木质化的带腋芽茎段经升汞消毒15 min,接种后暗培养5 d,MS+0.2 mg·L^-1 NAA培养基中添加3 mg·L^-1 PVP+2000 mg·L^-1 Vc或者6 mg·L^-1 PVP+3000 mg·L^-1 AC,可在低污染率的前提下将褐化率降为10%左右,一定程度上解决了茶树组培中褐化率高的难题;采用1/8 MS+1 mg·L^-1 IBA培养基有利于外植体生根且增殖迅速。本研究建立了十里香茶的无菌苗离体培养及再生体系,可通过组培手段对其进行大量繁殖。