Wheat cultivars and natural-based substances: Impacts on epidemiological parameters of yellow dwarf disease

For many years, control methods against aphid-transmitted barley/cereal yellow dwarf viruses (B/CYDV) in cereal fields were mainly based on the use of neonicotinoid (NNI)-coated seeds. The ban of NNI by the EU in 2018 has reinforced the interest of the scientific community in the characterization of genetic resources and biological protections to find alternatives to chemicals for the management of B/CYDV. Eleven BYDV-susceptible wheat varieties were tested using a set of experimental procedures to evaluate their potential to alter parameters linked to the biology of aphids (Rhopalosiphum padi) and spread of virus (BYDV-PAV). Moreover, two natural-based substances (azadirachtin and mineral oil) were tested for their impact on the ability of aphids to survive, colonize and transmit the virus. Results showed that the 11 genotypes tested have a level of susceptibility to virus infection similar to the susceptible reference cv. Rubisko. However, when characterization focused on virus load, latency period and aphid fecundity, partial resistance phenotypes were observed for some cultivars. Furthermore, azadirachtin increased aphid mortality and decreased aphid fecundity. Thus, in addition to genes described for their ability to limit B/CYDV infections, the genetic backgrounds of B/CYDV-susceptible wheat cultivars and azadirachtin-based treatment should be considered for future management strategies against yellow dwarf disease.     

大麦黄矮病毒GPV与GAV株系间异源装配现象的初步研究

 以我国麦区的大麦黄矮病毒GPV、GAV株系为材料,利用它们的蚜传特异性,将由禾谷缢管蚜传播的GPV和由麦长管蚜传播的GAV混合侵染到岸黑燕麦上,并进行继代传毒。混合侵染后代蚜传表现型的变化初步表明存在异源装配现象,且表现型混合发生的比例较高。用DAS-ELISA和RT-PCR法对混合侵染后代进行了测定,进一步证明了异源包装现象的存在。部分基因的核苷酸序列分析初步显示所测定的混合侵染后代中没有发生基因重组。     

感染大麦黄矮病毒的小麦中vsiRNA特征分析

 RNA沉默是植物中一种保守的抗病毒机制,其以大量病毒来源的小干扰RNA(virus-derived small interfering RNAs,vsiRNAs)的产生为标志,病毒在侵染寄主过程中可通过vsiRNAs靶向寄主转录本以对抗这种防御机制。BYDV-GAV引起的小麦黄矮病导致小麦黄化和矮化症状,对小麦生产构成严重威胁。通过深度测序技术,分析了感染BYDV-GAV的感病小麦品种‘小偃6号’中vsiRNAs的特征,共得到11 384个vsiRNAs,并预测到37 784个寄主靶基因。发现来源于BYDV-GAV基因组的正义和反义链的vsiRNAs的数量分布大致相等,在5’末端具有A和C偏好性,长度主要在21nt~22nt。靶基因的功能分析表明这些靶基因参与了广泛的生物学功能,尤其是在寄主-病原物互作中占的比重最大。选取25个参与寄主-病原互作的抗性相关基因进行定量验证,发现接种BYDV-GAV后有15个明显下调,6个上调,4个微弱下调,表明测序结果和靶基因预测可靠。推测BYDV-GAV可以通过vsiRNAs干扰寄主抗性基因表达和信号转导,从而实现对感病寄主的侵染。研究结果对揭示BYDV-GAV与小麦互作的分子机制具有重要意义。     

感染大麦黄矮病毒的小麦中vsiRNA特征分析

 RNA沉默是植物中一种保守的抗病毒机制,其以大量病毒来源的小干扰RNA(virus-derived small interfering RNAs,vsiRNAs)的产生为标志,病毒在侵染寄主过程中可通过vsiRNAs靶向寄主转录本以对抗这种防御机制。BYDV-GAV引起的小麦黄矮病导致小麦黄化和矮化症状,对小麦生产构成严重威胁。通过深度测序技术,分析了感染BYDV-GAV的感病小麦品种‘小偃6号’中vsiRNAs的特征,共得到11 384个vsiRNAs,并预测到37 784个寄主靶基因。发现来源于BYDV-GAV基因组的正义和反义链的vsiRNAs的数量分布大致相等,在5’末端具有A和C偏好性,长度主要在21nt~22nt。靶基因的功能分析表明这些靶基因参与了广泛的生物学功能,尤其是在寄主-病原物互作中占的比重最大。选取25个参与寄主-病原互作的抗性相关基因进行定量验证,发现接种BYDV-GAV后有15个明显下调,6个上调,4个微弱下调,表明测序结果和靶基因预测可靠。推测BYDV-GAV可以通过vsiRNAs干扰寄主抗性基因表达和信号转导,从而实现对感病寄主的侵染。研究结果对揭示BYDV-GAV与小麦互作的分子机制具有重要意义。     

人工饲料条件下大麦黄矮病毒GAV株系对麦长管蚜种群适合度的直接影响

植物病毒与介体蚜虫存在复杂的互作关系。前人关于植物病毒对蚜虫调控作用的研究主要集中在植物病毒通过寄主植物对蚜虫的间接影响上,未见植物病毒对介体蚜虫适合度直接调控的报道。鉴于此,我们以麦长管蚜Sitobion miscanthi (Takahashi)为试虫,以其传播的大麦黄矮病毒-GAV(Barley yellow dwarf virus GAV,BYDV-GAV)为测试病毒,以全纯人工饲料加入BYDV-GAV病毒提取液饲养麦长管蚜4 d,使之在不接触寄主植物条件下获毒,然后分别在全纯人工饲料和无毒小麦叶片上继续饲养,直至死亡。利用生命表技术分析麦长管蚜生长发育和繁殖参数。研究结果表明:在无毒小麦叶片饲养条件下,与未获毒对照麦长管蚜相比,获毒后麦长管蚜生活史参数成虫历期和产仔天数显著降低,繁殖力显著增加;种群参数内禀增长率、净繁殖率、周限增长率显著增加,平均世代周期显著降低。在全纯人工饲料条件下,与未获毒对照相比,获毒后麦长管蚜仅成虫历期和产仔天数显著下降,而其他生活史参数及种群参数均无显著差异。说明BYDV-GAV使得介体麦长管蚜在小麦叶片上的适合度显著提高,这是由麦长管蚜与寄主植物互作引起的,而病毒对介体麦长管蚜的适合度无直接调控作用。     

我国不同地区麦蚜携带BYDV-GAV比率的差异

为了研究我国不同地区麦蚜携带大麦黄矮病毒麦二叉蚜麦长管蚜非专化性株系(BYDV GAV)比率的差异,采用RT-PCR技术,对BYDV-GAV的传毒介体麦蚜带毒情况进行检测.所用方法具有较高的灵敏度和特异性,测定样本用量可少至1/200头蚜虫;对采自我国主要麦区的蚜虫样本进行分子检测,山西、甘肃、青海、陕西11个小麦黄矮病重病区蚜虫样本带毒率为56％～91.5％,而河北、河南两省4个非重病区蚜虫样本带毒率为2.5％～33％.通过试验证实,我国不同地区麦蚜携带BYDV-GAV比率存在差异,小麦黄矮病重病区山西、甘肃、青海、陕西等地的麦蚜带毒率高,而非重病区河北、河南等地的麦蚜带毒率低.     

大麦黄矮病毒GPV株系ORF4基因在大肠杆菌中的表达及特异抗血清的制备

 根据已报道的大麦黄矮病毒GPV株系(BYDV-GPV)相关基因序列设计合成引物,利用RT-PCR方法获得ORF4基因,并将其克隆到原核表达载体pET-5a中。经IPTG诱导、SDS-PAGE分析,结果表明:ORF4基因在大肠杆菌BL21(DE3) pLysS中获得了高效表达,分子量为17 kDa。以回收的表达产物为抗原免疫家兔,制备了BYDV-GPV 17kDa蛋白的特异性抗血清。Western blot检测结果,制备的抗血清可用于检测BYDV-GPV侵染后在燕麦体内表达的17 kDa蛋白。     

A comparative study of the Barley yellow dwarf virus species PAV and PAS: distribution,accumulation and host resistance

This study investigated the distribution and characteristics of the Barley yellow dwarf virus (BYDV) species BYDV‐PAS, which was recently separated from BYDV‐PAV, the most commonly studied BYDV species. Throughout 3 years of experimental monitoring of BYDV incidence, PAS was the most frequently occurring species infecting cereals and grasses in the Czech Republic. Furthermore, Rhopalosiphum maidis and Metopolophium dirhodum were recorded as BYDV‐PAS vectors, even though M. dirhodum does not usually transmit BYDV‐PAV. In field experiments with barley and wheat, where virus accumulation, symptoms and effect on the yield were tested, BYDV‐PAV was more severe than PAS. Infection with the BYDV‐PAV isolate resulted in greater expression of symptoms and also in a greater reduction in plant height and grain weight per spike than BYDV‐PAS. In a sensitive cultivar of barley (Graciosa), the amount of viral RNA of BYDV‐PAV was also significantly higher than that of BYDV‐PAS. In a tolerant line (Wbon‐123), however, no such differences were found. In conclusion, although BYDV‐PAS seems to be dominant in the Czech Republic, BYDV‐PAV has the potential to cause more significant crop losses in barley and wheat.     

BYDV PREDICTOR: a simulation model to predict aphid arrival, epidemics of Barley yellow dwarf virus and yield losses in wheat crops in a Mediterranean-type environment

BYDV PREDICTOR, a simulation model, was developed to forecast aphid outbreaks and Barley yellow dwarf virus (BYDV) epidemics in wheat crops in the grainbelt region of southwest Australia, which has a Mediterranean-type climate. The model used daily rainfall and mean temperature to predict aphid ( Rhopalosiphum padi ) buildup in each locality before the commencement of the cereal-growing season in late autumn, and to forecast the timing of aphid immigration into crops. The introduction of BYDV by aphid immigrants, aphid buildup within the crop, spread of BYDV, and yield losses were predicted for different sowing dates. The model simulations were validated with 10 years' field data from five different sites in the grainbelt, representing a wide range of scenarios. When first aphid arrival dates ranging from 1 June to 2 September were compared with predictions, 65% of the variation between sites and years was explained. Progress curves for the predicted percentage of plants infected with the serotype BYDV-PAV closely resembled the starting point and shape of those recorded in 14 out of 18 scenarios. Sensitivity analysis confirmed that the combination of a high proportion of immigrants vectoring BYDV, early sowing of crops and early start to aphid arrival relative to sowing date led to the most BYDV spread and greatest yield loss. The model was incorporated into a decision support system used by farmers in targeting sprays against aphids to prevent virus spread in autumn and winter. BYDV PREDICTOR could serve as a template for modelling similar virus/aphid vector pathosystems in other regions of the world, especially those with Mediterranean-type climates.     

青海省小麦种质资源对大麦黄矮病毒的抗性鉴定

为明确青海省不同小麦种质资源对大麦黄矮病毒(Barley yellow dwarf virus,BYDV)的抗性差异,于2014—2015年采用堆测法人工接种鉴定了178份种质资源的抗病性。结果显示,不同小麦种质资源对黄矮病的抗性存在较大差异,甘A100、川766、陕1059、兰麦-2的病情指数依次为14.83、23.60、23.99和24.66,表现出较好的抗病性;病情指数在50.00以上的高感品种有69份,包括尕老汉、白板麦、兴热密穗等,其中白板麦病情指数高达72.35,高于感病对照阿勃;其余种质资源病情指数在25.00~50.00之间,表现为感病;甘A101、甘A99、藏515、藏519、木汉麦、群科大白麦、拉胎板麦、小红麦-2和朗县折达25初期表现感病,后期恢复健康,有一定的耐病性;抗病性不同的种质资源感染BYDV后,对小麦产量的影响差异很大,抗病对照中4产量损失8.96%,耐病品种产量损失在13.64%~19.74%之间,高病品种尕老汉产量损失达41.91%,表明小麦种质资源中抗BYDV的品种极少。     

抗黄矮病小麦新品系的分子标记检测研究

本研究对以小麦-中间偃麦草异附加系L1和小麦-中间偃麦草部分双二倍体‘无芒中4’为抗源选育出的抗黄矮病小麦新品系进行分子检测和抗病性鉴定.通过应用RAPD、SSR、SCAR 3种分子标记OPF15、Xgwm37、SC-W37进行分子检测,并采用人工接种和大田自然感病的方式进行抗黄矮病鉴定,筛选到了‘93646’、‘2003-2’等高抗黄矮病的小麦新品系.分子检测抗黄矮病基因与田间抗病鉴定结果基本一致,应用的3种PCR标记都可以检测出抗病材料,但SCAR标记SC-W37特异性强、稳定性好,可在小麦抗黄矮病育种早代选择过程中发挥重要作用.     

6种病毒抑制剂对西藏青稞上大麦黄矮病的防治效果

通过田间小区试验,开展了6种病毒抑制剂对西藏青稞上大麦黄矮病防治效果的研究.采用带毒蚜虫人工接种法,于青稞起身拔节期每株接种10～15头带毒麦长管蚜,7 d后灭蚜,灭蚜后3d喷施病毒抑制剂,在成株期出现黄矮病症状后调查病株率、病情指数、防治效果,以及长势和产量.结果表明,6种病毒抑制剂对青稞上大麦黄矮病有不同程度的防治...     

大麦黄矮病毒PAV青海分离物运动蛋白抗血清制备及其与同属BYDVs的血清学关系

 大麦黄矮病毒(Barley yellow dwarf viruses, BYDVs)属于黄症病毒科,主要以蚜虫为介体进行传播,引发多种作物减产或绝收。本文将BYDV-PAV青海分离物的运动蛋白(MP)克隆到原核表达载体pDB-MBP-His上,转化大肠杆菌Rosetta(DE3),在IPTG诱导下表达蛋白分子量约为61 kDa的融合蛋白。将纯化后的蛋白作为抗原免疫‘新西兰’大白兔制备抗血清,Western blot检测本生烟瞬时表达带标签的MP蛋白,结果显示该抗血清效价为1∶32 000,灵敏度达1∶256,且该抗血清可与相差34个氨基酸的PAV015株系以及隶属于黄症病毒属其他的BYDVs(PAS、MAV、KerⅡ和KerⅢ)MP均能发生强烈的血清学反应,具有血清学相关性。本研究制备的大麦黄矮病毒PAV青海分离物运动蛋白多克隆抗体为探索BYDV-PAV运动蛋白的功能以及作用机制打下了基础。     

野茼蒿黄脉病毒的田间寄主范围及其基因多样性特征

为明确野茼蒿黄脉病毒(Crassocephalum yellow vein virus,CraYVV)的田间寄主范围及其群体基因结构特征,分别采用克隆和测序技术对采自云南省西双版纳傣族州、红河哈尼族彝族自治州的7种杂草和2种作物进行CraYVV的分离和鉴定,并通过生物信息学软件对分离到的CraYVV进行基因组结构、重组及基因遗传结构分析。结果显示,在赛葵、龙葵、臭牡丹、水茄、豨莶、野茼蒿和一点红7种杂草以及茄子和草莓2种作物中均分离到CraYVV,共获得20条CraYVV全长序列;CraYVV所有分离物分属2个株系,将其命名为YJ株系和JH株系;JH株系具有地理隔离特征,与YJ株系存在一定的遗传距离;重组分析显示,CraYVV是由烟草曲茎病毒(tobacco curly shoot virus,TbCSV)和云南烟草曲叶病毒(tobacco leaf curl Yunnan virus,TbLCYnV)重组产生;遗传结构分析显示,CraYVV中C1基因变异最显著,其次是C4基因和基因间隔区。表明CraYVV能侵染5科9种杂草和作物,具有较广的寄主范围,且菜豆金色花叶病毒属病毒能够侵染草莓、CraYVV能够侵染茄子,显示来源于杂草的菜豆金色花叶病毒属病毒在自然条件下能够侵染作物。     

Testing garlic cloves and bulblets for onion yellow dwarf virus by ACP-ELISA

Onion yellow dwarf virus (OYDV) was detected in cloves and aerial bulblets of garlic (Allium sativum) at levels as high as or higher than in leaves of plants grown from tested cloves. It is recommended to test bulblets or a few cloves per bulb before planting to determine if all cloves of a bulb are virus-free. This aids in early detection and allows a more thorough testing of stock than field testing.     

大麦黄矮病毒侵染小麦miRNA鉴定和特征分析

大麦黄矮病毒(barley yellow dwarf viruses, BYDVs)引起的小麦黄矮病严重威胁我国麦类生产,造成严重经济损失。植物中的miRNA调控植物生长发育、信号转导及对外界压力的反应,通过调控植物抗性基因的表达影响植物与病原物的互作。本研究对感染BYDV-GAV后3 d、7 d及健康对照的‘小偃6号’小麦样品进行miRNA测序,合并去冗余后分别得到99、96、95个已知的miRNA序列和806、809、1 024个新miRNA序列。对这些miRNA进行差异表达分析,BYDV-GAV侵染后3 d和7 d的小麦样品,与对照相比上调表达的miRNA数量分别为3个和7个,下调表达的为14个和12个。将差异表达的miRNA利用psRNATarget进行靶基因预测,共得到1 254个靶标基因。靶基因的KEGG和GO富集分析,进一步明确了其功能及作用通路。对14个病毒病症状相关的靶基因进行定量分析,结果表明随病毒侵染时间的延长,这些靶基因出现差异性表达,显示miRNA参与了寄主与病毒的互作。研究结果有助于揭示BYDV-GAV与寄主小麦的互作机理。     

大麦黄矮病毒侵染小麦miRNA鉴定和特征分析

 大麦黄矮病毒(barley yellow dwarf viruses,BYDVs)引起的小麦黄矮病严重威胁我国麦类生产,造成严重经济损失。植物中的miRNA调控植物生长发育、信号转导及对外界压力的反应,通过调控植物抗性基因的表达影响植物与病原物的互作。本研究对感染BYDV-GAV 后3 d、7 d及健康对照的‘小偃6号'小麦样品进行miRNA测序,合并去冗余后分别得到99、96、95个已知的miRNA序列和806、809、1 024个新miRNA序列。对这些miRNA进行差异表达分析,BYDV-GAV侵染后3 d和7 d的小麦样品,与对照相比上调表达的miRNA数量分别为3个和7个,下调表达的为14个和12个。将差异表达的miRNA利用psRNATarget进行靶基因预测,共得到1 254个靶标基因。靶基因的KEGG和GO富集分析,进一步明确了其功能及作用通路。对14个病毒病症状相关的靶基因进行定量分析,结果表明随病毒侵染时间的延长,这些靶基因出现差异性表达,显示miRNA参与了寄主与病毒的互作。研究结果有助于揭示BYDV-GAV与寄主小麦的互作机理。     

广西柑橘衰退病和黄脉病调查及其病原病毒的遗传多样性分析

为明确柑橘衰退病毒(citrus tristeza virus, CTV)和柑橘黄脉病毒(citrus yellow vein clearing virus, CYVCV)在广西柑橘上的发生?分布及其遗传变异情况, 于2020年至2021年对百色?北海?崇左?贵港?桂林?河池?贺州?来宾?柳州?南宁?梧州和玉林等12个柑橘产区进行了病毒病调查?采用RT-PCR对采集样品进行了病毒检测, 并基于病毒分离物外壳蛋白(coat protein, CP)基因的核苷酸序列进行比对分析, 构建系统发育树?结果表明:采集的737份柑橘样品中, CTV的检出率为20.62%, CYVCV检出率为18.32%, CTV的检出率略高于CYVCV?病毒复合侵染的现象在采集的柑橘样品中普遍存在, CTV和CYVCV复合侵染率高达34.50%?对RT-PCR产物测序共获得12个CTV分离物和6个CYVCV分离物的CP基因序列?遗传多样性分析发现, CTV和CYVCV的CP基因序列都较保守, CTV分离物的遗传进化与地理来源?寄主来源均没有明显相关性, 但CYVCV分离物的遗传进化与地理位置具有相关性, 而与寄主来源无明显相关性?上述研究结果可为深入了解CTV和CYVCV在广西的流行情况以及柑橘病毒病的检疫和防控提供参考?