瓜果上主要实蝇类害虫PCR-RFLP分子鉴定技术的建立及应用

[目的]建立一种针对实蝇幼虫快速鉴定的技术方法,并应用该技术以明确我国瓜果上主要实蝇类害虫的优势种.[方法]以我国瓜果蔬菜上危害最为严重的瓜实蝇Bactrocera cucurbitae(Coquillett)、南亚果实蝇Bactrocera tau Walker和橘小实蝇Bactrocera dorsalis(Hendel)为对象,基于3种实蝇种间存在线粒体细胞色素氧化酶亚基Ⅰ因子(mtCOⅠ)基因的序列差异,利用3种限制性内切酶SapⅠ、BsmⅠ和Sac Ⅰ酶切识别这些差异序列的特异性酶切位点,根据酶切后mtCOⅠ基因片段大小不同,对3种实蝇进行快速的鉴定识别,并进一步应用此技术对来自我国7省11市的7种瓜菜和2种水果上的21份实蝇幼虫样本进行鉴定.[结果]通过单独或组合酶切的方式建立了一套针对瓜实蝇、南亚果实蝇和橘小实蝇的PCR-RFLP快速分子鉴定技术,并将此方法应用于田间监测.田间鉴定结果表明,湖北、湖南、浙江等地区危害瓜菜的实蝇主要是南亚果实蝇,广东、广西、海南等地区瓜菜上的实蝇种类主要是瓜实蝇,且瓜实蝇和南亚果实蝇存在寄主重叠现象,云南危害火龙果和芒果等水果上的实蝇主要是橘小实蝇.[结论]本研究建立的瓜实蝇、南亚果实蝇和橘小实蝇的识别鉴定方法,因不受虫体、虫态的影响,且具有低成本、高效、快速、重复性好的特点,适用于田间瓜果上实蝇样本的快速诊断检测.依据田间鉴定结果,预测近年来瓜实蝇可能有逐渐向北扩散的趋势.     

我国主要瓜类实蝇的生物防治研究进展

本文概述了国内外主要瓜类实蝇的生物防治研究进展,概括了瓜实蝇Bactrocera cucurbitae和南亚果实蝇Bactrocera tau的主要自然天敌类群,论述了植物源提取物、性信息素、化学物质、诱集植物和不育技术在防治瓜类实蝇方面的应用,并讨论了生物防治存在的问题和今后持续控制瓜类实蝇方面的发展趋势。     

云南西部实蝇的多样性

2005和2006年,在云南西部瑞丽、潞江坝、六库、保山、大理5个地区,采用信息素引诱法及被害瓜果内实蝇收集法对5个地区实蝇多样性进行分析.结果表明,5个地区实蝇丰富度的发生模式明显分为2种类型:瑞丽和潞江坝实蝇全年发生;六库、保山和大理实蝇季节性发生;瑞丽的实蝇种类丰富度最高,有20种,其次是潞江坝,11种,而保山、大理的实蝇种类丰富度低,分别为4种和3种;该区域实蝇优势种为桔小实蝇(Bactrocera dorsalis)和南亚果实蝇(B. tau);2年实蝇多样性指数均以六库最低,以潞江坝最高.各地实蝇种类相似性变化较大, 相似性系数在0.15～1.00 之间.结果为了解云南西部实蝇种类分布以及预防实蝇危害提供了基础数据.     

离腹寡毛果实蝇性别决定基因transformer 2及其在昆虫中的进化

对离腹寡毛果实蝇性别决定基因transformer 2(tra-2)进行多角度的进化分析,为研究离腹寡毛果实蝇性别决定分子机制奠定基础。通过PCR结合RACE技术克隆瓜实蝇Bactrocera cucurbitae同源transformer 2基因的c DNA全长和内含子序列,并对该基因进行序列和进化分析。克隆得到瓜实蝇tra-2基因c DNA全长1467 bp,开放读码框753 bp,编码250个氨基酸残基,Gen Bank登录号为KR056217。瓜实蝇tra-2基因与其他6种离腹寡毛果实蝇的基因结构相似,均包含8个外显子和7个内含子。7种离腹寡毛果实蝇与实蝇科的地中海实蝇和按实蝇、蝇科的家蝇和丽蝇科的铜绿蝇为单一起源,而果蝇Tra-2表现出明显的分异。分子进化分析结果表明tra-2基因核苷酸变异主要来源于同义变异,接受净化选择压力。本研究明确了瓜实蝇tra-2的c DNA和基因组DNA结构特征,离腹寡毛果实蝇tra-2蛋白高度保守,主要接受净化选择压力。     

福州实蝇监测初报

自2005年5月29日至2006年5月28日在福州金山福建农林大学校园内进行了实蝇监测。采用甲基丁香酚(M e)、诱蝇酮(Cue)和地中海实蝇诱芯(T),诱集到橘小实蝇[Bactrocera(Bactrocera)dorsalis]、瓜实蝇[B.(Zeugodacus)cucurbi-tae]、南瓜实蝇[B.(Zeugodacus)tau]和具条实蝇(B.scutellata)4种实蝇,但未诱到地中海实蝇(Ceratitis capitata)。其中,橘小实蝇诱集量最大,8月中下旬达到高峰。本研究为掌握福州地区实蝇发生动态提供了基础资料。     

橘小实蝇、瓜实蝇和南亚果实蝇人工饲料的优化

在温度25²8℃、相对湿度70%28℃、相对湿度70%⁷5%和光照周期L∶D=14∶10条件下进行了人工大量饲养橘小实蝇Bactrocera dorsalis(Hendel)和瓜实蝇B.cucurbitae(Coquillett)成虫人工饲料配方的筛选试验。结果显示,1∶2重量比例混合的蔗糖和啤酒酵母是饲养这2种果实蝇的最佳成虫人工饲料,用其饲养的单雌产卵量、产卵期和孵化率分别为424.1675%和光照周期L∶D=14∶10条件下进行了人工大量饲养橘小实蝇Bactrocera dorsalis(Hendel)和瓜实蝇B.cucurbitae(Coquillett)成虫人工饲料配方的筛选试验。结果显示,1∶2重量比例混合的蔗糖和啤酒酵母是饲养这2种果实蝇的最佳成虫人工饲料,用其饲养的单雌产卵量、产卵期和孵化率分别为424.16⁴45.75粒,30.90445.75粒,30.90³1.87 d,74.60%31.87 d,74.60%⁷5.40%。同时,对18种由不同配方配制成的幼虫人工饲料饲养橘小实蝇、瓜实蝇和南亚果实蝇B.tau(Walker)的效果进行了比较。结果表明,玉米和麦麸作为饲料的介质优于麦片和麦麸。橘小实蝇幼虫人工饲料的优化配方为:玉米+麦麸(125 g+25 g),蔗糖25 g,啤酒酵母25 g,对羟基苯甲酸甲酯0.9 g,1 mol/L盐酸4 mL,纸巾4 g,自来水300 mL;用其幼虫人工饲料饲养该虫的生物学参数包括子代孵化率、化蛹率、羽化率和平均蛹重分别为81.17%±0.05%,96.41%±0.02%,94.85%±0.01%与(19.40±0.08)mg。而瓜实蝇和南亚果实蝇幼虫人工饲料的优化配方为:玉米+麦麸(100 g+50 g),蔗糖30 g,啤酒酵母25 g,以及一定量的其他组分(同上);用其饲料饲养这2种果实蝇的相关参数:子代孵化率、化蛹率、羽化率和平均蛹重分别为78.50%±0.04%与76.96%±0.12%,95.73%±0.03%与94.69%±0.02%,94.57%±0.02%与95.82%±0.03%,(18.62±0.23)mg与(22.83±1.38)mg。试验证实,优化后的成、幼虫人工饲料具有饲养效果好、方法简便,配方材料来源广泛和价格低廉等优点,可用于室内人工大量饲养上述3种果实蝇属害虫。     

基于mtDNA cox1和cox2基因的宽带果实蝇种群遗传结构分析

【目的】宽带果实蝇Zeugodacus scutellatus (Hendel)是我国分布范围最广的实蝇类昆虫之一,掌握其入侵来源以及传播扩散模式是害虫防控的基本前提。【方法】本研究基于线粒体DNA cox1和cox2基因片段,对我国宽带果实蝇的种群遗传结构进行了初步研究,并以此为依据对其入侵扩散过程进行了分析。【结果】我国东部和西部宽带果实蝇种群单倍型多样性(h)、核苷酸多样性(π)、遗传分化系数(F_(st))、基因流(N_m)均高于中部地区,日本宽带果实蝇种群单倍型数量、单倍型百分比、遗传多样性等参数均高于我国宽带果实蝇种群。中性检验表明,宽带果实蝇在我国经历过快速扩张事件,地理隔离在种群遗传结构中并未产生关键作用。【结论】上海和重庆可能是其最早进入我国的地区,该虫可能经由两条主要路径实现其在我国大范围扩张:一条直接通过港口贸易登陆上海,在长三角地区传播扩散,依此基础沿长江水系迅速向内陆扩张;另一路则以重庆为起点,在云贵甘陕等西部地区迅速扩张,并在此基础上借助长江沿线适宜的定殖区向南北传播扩散。     

温度、光照强度对瓜实蝇成虫飞行行为的影响

【背景】瓜实蝇是我国南方瓜果的主要害虫。本试验分析了影响瓜实蝇成虫飞行行为及活跃度的原因,以便能更有效地控制瓜实蝇危害。【方法】在已知瓜实蝇的生物学特性及部分活动规律的基础上利用智能人工气候箱等设施,分别测定温度、光照对瓜实蝇的飞行行为的影响。【结果】瓜实蝇成虫在15℃下开始飞行,飞行活动的群体数量随着温度的升高而增加,最适飞行活动温度范围为20~30℃,低温10℃明显对其飞行行为有抑制作用,高温(35℃以上)明显对其有刺激作用,促进其飞行行为的发生。瓜实蝇成虫的活跃度在25~40℃下较高,均大于50%,在10、15和45℃下则为0。瓜实蝇成虫在2400 lx光照条件下开始飞行活动,飞行活动的群体数量随着光照强度的升高而增强,最适飞行活动的光照为9600 lx,但高强度光照12000 lx对其飞行行为有抑制作用。活跃成虫的数量随着光照强度的升高而增加。【结论与意义】温度、光照强度过高或过低对瓜实蝇的飞行行为都有抑制作用,适度的高温有促进作用。     

基于mtDNA COI基因序列的中国柑橘果实蝇的分子鉴定及柑橘大实蝇种群的遗传多样性分析

【目的】探明危害我国柑橘的实蝇种类以及柑橘大实蝇Bactrocera minax不同地理种群和不同寄主种群的遗传多样性。【方法】利用mtDNA COI基因对危害柑橘的果实蝇进行种类鉴定,采用MEGA软件对其中28个地理种群的535头果实蝇COI基因片段(约505 bp)序列进行比对,分析种间及种内遗传距离,构建系统发育树。使用DnaSP软件分析柑橘大实蝇不同地理种群和不同寄主种群的遗传多样性。【结果】从柑橘虫果内共鉴定出4种实蝇,分别为柑橘大实蝇B.minax、桔小实蝇B.dorsalis、蜜柑大实蝇B.tsuneonis和瑞丽果实蝇B.ruiliensis。这4种实蝇的种间遗传距离为0.0264～0.2410,种内遗传距离为0.0000～0.0140,种间与种内遗传距离没有重叠区域。单个柑橘虫果内一般仅有1种实蝇,极个别柑橘果实可同时被两种实蝇危害(4/43);在这些为害柑橘的实蝇种类中,以柑橘大实蝇的个体数量比例最大,占90.70%。柑橘大实蝇地理种群遗传多样性高,28个种群共有17个单倍型。【结论】柑橘大实蝇是所调查地区柑橘实蝇的绝对优势种,其种群遗传分化程度较高,扩散危害风险大。本研究结果对柑橘果实蝇类的监测和防控具有重要意义。     

寄主植物对南亚果实蝇过冷却点和体液冰点的影响

南亚果实蝇Zeugodacus tau(Walker)是一种世界性的重要检疫害虫,严重危害多种水果和瓜菜。为明确寄主植物对南亚果实蝇耐寒性的影响,本文利用过冷却点仪,对取食苦瓜Momordica charantia L.、黄瓜Cucumis sativus L.、丝瓜Luffa aegyptiaca Miller和杧果Mangifera indica L.的南亚果实蝇5个发育阶段的过冷却点和体液冰点进行了测定。结果显示：每一组寄主植物上的5个发育阶段中,南亚果实蝇5日龄蛹的过冷却点和体液冰点均为最低,显著低于1日龄蛹、幼虫和成虫;不同寄主南亚果实蝇同一发育阶段的过冷却点和体液冰点存在显著差异,均值分别为-23.97～-8.06℃和-14.02～-0.59℃,其中取食苦瓜的南亚果实蝇5日龄蛹过冷却点和体液冰点最低,均值分别为-23.97℃和-14.02℃,但与黄瓜无显著差异;4种寄主中老熟幼虫、1日龄蛹、1日龄雌成虫和5日龄雌成虫过冷却点均为取食黄瓜最低;杧果种群1日龄雌成虫过冷却点最高,均值为-7.47℃,苦瓜种群老熟幼虫体液冰点最高,均值为-0.59℃。本研究有助于了解寄主对南亚果实...     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

Comparative morphology of the carpel in the Liliaceae: Hewardieae, Petrosavieae, and Tricyrteae

The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.     

Enterovirus 71 3C proteolytically processes the histone H3 N-terminal tail during infection

Highlights
1. The N-terminal tail of histone H3 is specifically cleaved during EV71 infection.
2. Viral protease 3C is identified as a protease responsible for proteolytically processing the N-terminal H3 tail.
3. Our finding reveals a new epigenetic regulatory mechanism for Enterovirus 71 in virus-host interactions.     

Detection of EBV and HHV6 in the Brain Tissue of Patients with Rasmussen’s Encephalitis

Rasmussen’s encephalitis (RE) is a rare pediatric neurological disorder, and the exact etiology is not clear. Viral infection may be involved in the pathogenesis of RE, but conflicting results have reported. In this study, we evaluated the expression of both Epstein-Barr virus (EBV) and human herpes virus (HHV) 6 antigens in brain sections from 30 patients with RE and 16 control individuals by immunohistochemistry. In the RE group, EBV and HHV6 antigens were detected in 56.7% (17/30) and 50% (15/30) of individuals, respectively. In contrast, no detectable EBV and HHV6 antigen expression was found in brain tissues of the control group. The co-expression of EBV and HHV6 was detected in 20.0% (6/30) of individuals. In particular, a 4-year-old boy had a typical clinical course, including a medical history of viral encephalitis, intractable epilepsy, and hemispheric atrophy. The co-expression of EBV and HHV6 was detected in neurons and astrocytes in the brain tissue, accompanied by a high frequency of CD8⁺ T cells. Our results suggest that EBV and HHV6 infection and the activation of CD8⁺ T cells are involved in the pathogenesis of RE.     

Perinatal Vertical Transmission of Chikungunya Virus in Ruili,a Town on the Border between China and Myanmar

正Dear Editor,Chikungunya virus (CHIKV), an arbovirus in the family of Togaviridae, genus Alphavirus, is transmitted by the A.aegyptii or A. albopictus mosquito, and causes disease in humans characterized by fever, rash, and arthralgia (Silva and Dermody 2017; Suhrbier 2019). It was first reported in 1953 in Tanzania, and caused only a few outbreaks and sporadic cases in Africa and Asia in last century. However, in the epidemic in 2004, CHIKV acquired mutations that conferred enhanced transmission by the A. albopictus mosquito(Schuffenecker et al. 2006). Since then, it has successively caused outbreaks in Africa, the Indian Ocean, South East Asia, the South America, and Europe (Zeller et al. 2016).     

Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2

In conclusion, the novel visual RT-LAMP assay is a simple, rapid, and sensitive approach for detection of SARS-CoV-2, and it is ready for application in primary care and community hospitals or health care centers, and even patients' own houses in response to the current SARS-CoV-2 epidemic because the assay does not require sophisticated equipment and skilled personnel. Furthermore, it is also ready to be used in fields for screening samples from wild animals and environments to facilitate the identification of potential intermediate hosts that mediate the cross-species transmission of SARS-CoV-2 from bats to humans.