甘薯茎腐病症状及其病原鉴定

 甘薯茎腐病是危害甘薯的一种严重病害,甘薯茎、叶柄、叶片和块根均可被害。该病典型症状是茎基部发黑和变软腐烂,叶发黄,茎和块根维管束黑褐色,引起块根腐烂、有臭味。通过对甘薯茎腐病典型症状样本的采集、病原菌的分离和纯化以及致病性测定,明确该病害是一种细菌病害。通过对甘薯茎腐病菌的菌体形态和培养特性观察发现,病原菌是革兰氏阴性细菌,菌体短杆状,大小约为2.36 μm×0.4 μm, 周生鞭毛,可在烟草上激发过敏性反应(HR)。Biolog测定、脂肪酸分析、16S rDNA序列分析、MALDI-TOF质谱鉴定和8个看家基因(dnaX、rplB、fusA、gapA、gyrA、purA、recA和rpoS)的序列系统发育分析,发现该病原菌与达旦提狄克氏菌Dickeya dadantii高度一致。这些结果说明,浙江省发生的小番薯病害是甘薯茎腐病,病原为D. dadantii。     

黄岩甘薯茎基部腐烂病发生情况与防控措施

为有效控制浙江省台州市黄岩区甘薯茎基部腐烂病的发生为害,2015-2020年研究了该病在黄岩的发生情况、影响该病的因素以及防控病害的措施,明确了引起黄岩西部山区甘薯茎基部腐烂的病原菌包括甘薯茎腐病菌(Dickeya dadantii)、甘薯基腐病菌(Phomopsis destruens)、甘薯根腐病菌(Fusariu...     

河北省甘薯茎腐病研究初报

甘薯茎腐病是近几年在我国发现的一种新的细菌性病害,目前该病已在我国的福建、广东、江西、广西、海南、河南、重庆、江苏等地发生。2013年10月在对河北省甘薯病害的调查中在文安县发现了大量疑似甘薯茎腐病的病株,给当地的甘薯生产造成了严重的影响。经对疑似病株病原菌的分离纯化、柯赫氏法则验证、形态观察以及基于16SrDNA序列的分析,最终确定该病害为甘薯茎腐病,病原菌为达旦提狄克氏菌(Dickeya dadantii)。这是该病害首次在河北省被发现。     

甘薯对马铃薯腐烂茎线虫趋化性的影响

由马铃薯腐烂茎线虫侵染造成的甘薯茎线虫病是一种严重影响甘薯产量和品质的病害。线虫向寄主根部移动及取食危害可能是受寄主释放的化感信息物质控制。为最终明确甘薯中对线虫具有高效引诱能力的物质,本研究利用水琼脂平板法测试了甘薯不同品种与不同组织对线虫的引诱作用。结果表明不同品种甘薯薯块和薯茎对马铃薯腐烂茎线虫引诱能力具有一定差异,感线虫品种栗子香的薯块对线虫的引诱率要高于其他测试品种。随着薯块/薯茎在水琼脂平板上放置时间延长,其吸引线虫的效果增加,96 h后引诱率达到20%。同一品种甘薯茎的引诱能力显著高于薯块、薯叶。甘薯中所含对线虫具引诱作用的活性物质对热稳定,80℃加热8 h后薯茎对线虫引诱率与新鲜茎相比无显著差异,为4.5%。甘薯茎经过不同有机溶剂分别萃取后,乙醇萃取物对线虫的引诱率为6.7%,显著高于其他萃取物。本文初步明确了甘薯中所含线虫趋化物的性质,为其开发应用奠定了基础。     

中国发现甘薯长喙壳菌引起枣子果实腐烂

 2013年8月,在中国昆明发现一种枣子果实的腐烂病。通过致病性测试,证明从发病组织分离出的病菌能引起枣子果实腐烂。经过形态学和分子鉴定,确定该病原菌为甘薯长喙壳菌。这是在中国首次报道由甘薯长喙壳菌引起的枣子果实腐烂病。     

不同药剂防治甘薯腐烂茎线虫病试验研究

甘薯腐烂茎线虫病是甘薯上的主要病害。为筛选出有效药剂,2015～2016年田间试验了7种药剂穴施对甘薯腐烂茎线虫的防治效果及其对甘薯成活率和产量的影响。结果表明,30%辛硫磷微囊悬浮剂、5%辛硫磷颗粒剂、30%三唑磷微囊悬浮剂和10%噻唑磷颗粒剂对甘薯腐烂茎线虫有很好的防治效果。其中30%辛硫磷微囊悬浮剂、30%三唑磷微囊悬浮剂不仅对甘薯的存活率和生长无不良影响,而且增产效果相对较高,可作为防治甘薯腐烂茎线虫病的主推药剂。     

警惕花生黑腐病菌的入侵

2009年在我国广东省发现了花生黑腐病,该病害的典型症状是花生茎基部、果针和豆荚以及整个根系变黑腐烂,在温暖潮湿条件下,罹病部位产生大量红色小点,是病原菌的子囊壳。病原菌为冬青丽赤壳菌（Calonectria ilicicola Boedijin &; Reitsma）,无性阶段是寄生柱枝孢（Cylindrocladium parasiticum Crous, Wingfield &; Alfenas）。花生黑腐病菌可侵染花生和大豆等20多种作物。应警惕花生黑腐病菌的入侵,及时有效地防止病害蔓延。     

马铃薯腐烂茎线虫RPA-LFD可视化快速检测方法的建立

 马铃薯腐烂茎线虫是为害我国甘薯和马铃薯的一种重要植物病原线虫,也是我国重要的检疫性有害生物。为实现对该线虫的准确、快速且可视化的检测,本研究以马铃薯腐烂茎线虫rDNA-ITS序列为靶标构建了重组酶聚合酶结合侧流层析试纸条(RPA-LFD)的可视化快速检测体系。该体系可在39 ℃条件下15 min内特异性地完成对马铃薯腐烂茎线虫的检测,对A型(甘薯种群)和B型(马铃薯种群)单头线虫(J4)的检出底限均为3 125^-1头线虫,可以直接对土壤和甘薯茎中的马铃薯腐烂茎线虫进行检测,灵敏度可达1头(J4)/10 g土壤和1头(J4)/2 g甘薯茎组织。该体系操作简便、成本低廉、结果可视,可为马铃薯腐烂茎线虫的早期预警和口岸检疫提供技术支撑。     

甘薯茎线虫rDNA-ITS1区的PCR扩增与序列分析

 利用PCR技术获得了甘薯茎线虫rDNA-ITS1区序列。序列分析表明,采自我国河北、山东、安徽的甘薯茎线虫16个地理种群的ITS1区序列分化为短型(S)和长型(L)2种基因型。山东费县芍药山乡4个地理种群为L型,ITS1区长度为466 bp;河北、安徽和山东费县新庄乡的12个地理种群为S型,ITS1区长度为288 bp。甘薯茎线虫与鳞球茎茎线虫(Ditylenchus dipsaci)的rDNA-ITS1序列同源性为52.0%~52.5%,与腐烂茎线虫(D.destructor)序列同源性为82.0%~85.4%;我国甘薯茎线虫不同地理种群间的序列同源性为96.6%~100.0%。     

基于Dickeya dadantii特有标志基因检测甘薯茎腐病菌

 甘薯细菌性茎腐病是由达旦提狄克氏菌(Dickeya dadantii)引起的一种检疫性病害,近年来在我国多地发生,严重威胁我国甘薯产业的发展。建立特异灵敏的检测D. dadantii的方法对于鉴定检疫病原菌、田间监测病原菌和防控病害有重要意义。本研究对Dickeya属菌株的全基因组序列进行比较基因组学分析,筛选到D. dadantii特有的标志基因,针对标志基因设计引物,其中针对编码登录号为WP_077245517未知蛋白基因的1对引物Dad1-F(5′-CATATCAACCAGACCAGCCGTT-3′)和Dad1-R(5′-CGGCCTGCTTTTAAACAACGTATTA-3′)能只从D. dadantii扩增到167 bp目的片段。由此建立了特异灵敏的常规PCR和实时荧光定量PCR检测D. dadantii的方法,为鉴定检疫甘薯茎腐病病原菌和田间监测病害提供了有效方法。     

Interactions between a crinivirus, an ipomovirus and a potyvirus in coinfected sweetpotato plants

Novel and severe symptoms of chlorosis, rugosity, leaf strapping and dark green islands, designated as sweetpotato severe mosaic disease (SPSMD), were caused by dual infection of Sweet potato mild mottle virus (SPMMV; Ipomovirus ) and Sweet potato chlorotic stunt virus (SPCSV; Crinivirus ) in three East African sweetpotato cultivars (Tanzania, Dimbuka and New Kawogo). The storage root yield was reduced by ∼80%, as compared with healthy plants under screenhouse conditions in Uganda. Plants infected with SPMMV or SPCSV alone showed nonsignificant or 50% yield reduction, respectively. SPCSV reduced resistance to SPMMV in sweetpotato, similar to the situation with resistance to Sweet potato feathery mottle virus (SPFMV; Potyvirus ) that breaks down following infection with SPCSV, followed by development of sweet potato virus disease (SPVD). In single virus infections with SPMMV and SPFMV or their coinfection, cvs Tanzania and Dimbuka were initially systemically infected, displayed symptoms and contained readily detectable virus titres, but new leaves were symptomless with very low virus titres, indicating recovery from disease. In contrast, cv. New Kawogo remained symptomless and contained low SPMMV and SPFMV titres following graft inoculation. These moderate and high levels of resistance to SPMMV and SPFMV, respectively, were lost and cultivars succumbed to a severe disease following coinfection with SPCSV. The synergistic interactions increased titres of SPMMV and SPFMV RNA by ∼1000-fold as quantified by real-time PCR, whereas SPCSV titres were reduced twofold, indicating an antagonistic interaction. Coinfection with SPMMV and SPFMV caused no detectable changes in virus titres or symptom severity.     

土壤中烟嘧磺隆和莠去津残留对甘薯的药害评价

为了明确玉米田除草剂烟嘧磺隆和莠去津土壤残留对后茬作物甘薯生长的影响,于2017年分别在河北省保定满城区、文安县、霸州市和石家庄藁城区采集甘薯田土壤样品,采用QuEChERS法,结合超高效液相色谱-串联质谱法 (UPLC-MS/MS) 测定了土壤中烟嘧磺隆和莠去津的残留量;通过盆栽试验,测定了烟嘧磺隆、莠去津以及二者混用对甘薯的药害作用。结果表明:在甘薯药害表现严重的河北保定满城区、文安县、霸州市3地甘薯田土壤中烟嘧磺隆的残留量在11.80~87.97 μg/(kg土) 之间,莠去津残留量在2.71~8.40 μg/(kg土) 之间;而在石家庄藁城区无药害症状甘薯田土壤中只检测到莠去津,含量在3.43~4.42 μg/(kg土) 之间。盆栽试验结果表明,栽后20 d每千克甘薯田土壤中烟嘧磺隆含量大于1 μg时,甘薯产生药害症状;烟嘧磺隆对甘薯根系IC₅₀值为3.74 μg/(kg土),IC₁₀值为1.27 μg/(kg土)。土壤中莠去津含量在2~40 μg/(kg土) 范围内,对甘薯茎叶和根系没有可见药害症状,但对根系生长具有轻微抑制作用。烟嘧磺隆与莠去津按质量比10 : 1、5 : 1、5 : 2和5 : 5混用对甘薯药害有一定的拮抗作用,共毒系数CTC₅₀值分别为66.25、59.16、61.30和79.47。烟嘧磺隆对甘薯药害症状主要表现为植株矮小,叶片小,整株或心叶黄化,根系发育受阻,侧根短粗。生产上造成甘薯药害的诱因是土壤中残留的烟嘧磺隆,对甘薯药害阈值为1 μg/(kg土)。     

Two Serotypes of Sweetpotato feathery mottle virus in Uganda and Their Interaction with Resistant Sweetpotato Cultivars

ABSTRACT Isolates of Sweetpotato feathery mottle virus (SPFMV, genus Potyvirus, family Potyviridae) were obtained in several districts of Uganda from sweetpotato plants infected with the sweetpotato virus disease (SPVD), the most important disease of this crop in Africa. A monoclonal antibody (MAb 7H8) raised against the coat proteins (CP) of a mixture of the SPFMV strain C (United States) and the isolate SPV-I (West Africa) distinguished Ugandan SPFMV isolates into those detectable and not detectable by the MAb. These two serotypes differed in prevalence in different districts of Uganda and in two common sweetpotato cultivars. Both serotypes could be transmitted simultaneously by single aphids. The serotypes differed in their ability to systemically coinfect sweetpotatoes that were infected with Sweetpotato chlorotic stunt virus (SPCSV, genus Crinivirus), the virus required to induce SPVD in SPFMV-infected plants. One sweetpotato breeding line, resistant to SPFMV from the New World, was infected by graft-inoculation with all SPFMV isolates from Uganda. Another SPFMV-resistant sweetpotato line became infected with SPFMV and developed SPVD only following coinoculation with SPCSV.     

甘薯根腐病的初步研究

 甘薯根腐病是我国新发生的一种病害。近年,在甘薯主要产区蔓延迅速,致使产量遭受严重损失。     

Natural wild hosts of sweet potato feathery mottle virus show spatial differences in virus incidence and virus-like diseases in Uganda

Sweet potato feathery mottle virus (SPFMV, genus Potyvirus) is globally the most common pathogen of sweetpotato. An East African strain of SPFMV incites the severe 'sweetpotato virus disease' in plants co-infected with Sweet potato chlorotic stunt virus and threatens subsistence sweetpotato production in East Africa; however, little is known about its natural hosts and ecology. In all, 2,864 wild plants growing in sweetpotato fields or in their close proximity in Uganda were observed for virus-like symptoms and tested for SPFMV in two surveys (2004 and 2007). SPFMV was detected at different incidence in 22 Ipomoea spp., Hewittia sublobata, and Lepistemon owariensis, of which 19 species are new hosts for SPFMV. Among the SPFMV-positive plants, approximately 60% displayed virus-like symptoms. Although SPFMV incidence was similar in annual and perennial species, virus-like diseases were more common in annuals than perennials. Virus-like diseases and SPFMV were more common in the eastern agroecological zone than the western, central, and northern zones, which contrasted with known incidence of SPFMV in sweetpotato crops. The data on a large number of new natural hosts of SPFMV detected in this study provide novel insights into the ecology of SPFMV in East Africa.     

甘薯根腐病抗性在不同环境条件下的表现及遗传趋势

结果表明,甘薯根腐病发病轻重与环境条件有很大关系,表现为干旱少雨的年份发病较重,降雨量较多的年份发病较轻;土壤瘠薄的发病较重,肥力条件较好的则发病较轻;通常情况下,年份间品种的抗性表现较为一致,但遇特殊气候则年份间品种的抗性有一定的差异。对1150份甘薯品种资源及育种材料的根腐病抗性鉴定结果表明,高抗型材料占14.6％,抗病型占15.7％,感病型占26.0％,高感型占43.7％。对754份材料及亲本的抗性分析表明,不同的抗性组合后代中均可分离出高抗至高感类型的材料,杂交后代的抗性强弱随双亲抗性水平的提高而提高。中国自1970年以来采用品种间杂交和种间杂交育种技术,先后育成了一批高产、优质的高抗型优良品种。     

Development of a Real-Time Polymerase Chain Reaction Assay for Quantifying Verticillium albo-atrum DNA in Resistant and Susceptible Alfalfa

ABSTRACT A precise real-time polymerase chain reaction (PCR) assay was developed for quantifying Verticillium albo-atrum DNA. The assay was used in a repeated experiment to examine the relationship between the quantity of pathogen DNA detected in infected leaves and shoots and the severity of Verticillium wilt symptoms in several alfalfa cultivars expressing a range of disease symptoms. Plants were visually inspected for symptoms and rated using a disease severity index ranging from 1 to 5, and the quantity of pathogen DNA present in leaves and stems was determined with real-time PCR. No significant differences in pathogen DNA quantity or disease severity index were observed for experiments or for cultivar-experiment interactions. Significant differences were observed between cultivars for the quantity of pathogen DNA detected with real-time PCR and also for disease severity index ratings. In both experiments, the highly resistant check cultivar Oneida VR had significantly less pathogen DNA, and significantly lower disease severity index ratings than the resistant cultivar Samauri, the moderately resistant cultivar Vernema, and the susceptible check cultivar Saranac. In both experiments, the Spearman rank correlation between the amount of V. albo-atrum DNA detected in leaves and stems with real-time PCR and disease severity index ratings based on visual examination of symptoms was positive (>0.52) and significant (P < 0.0001). These results suggest that resistance to Verticillium wilt in alfalfa is characterized by a reduced colonization of resistant genotypes by the fungus.     

Degeneration in sweetpotato due to viruses,virus‐cleaned planting material and reversion: a review

This review examines viral degeneration in sweetpotato in different regions of the World, particularly that caused by Sweet potato chlorotic stunt virus (SPCSV) and Sweet potato feathery mottle virus (SPFMV), comparing impacts on yield in single and complex infections of all the major viruses affecting the crop. How cultivars are generated and virus resistance are also covered, especially for Africa. The synergistic (SPCSV + SPFMV) sweet potato virus disease (SPVD) is amongst the most dramatic diseases of sweetpotato but its overall yield impacts may not be as high as is generally assumed. It is constrained by resistance, roguing and selection of symptomless planting material. Instead, the cumulative impact of individual and combinations of symptomless viruses may be globally greater. These include sweepoviruses and various potyviruses, of which the commonest is SPFMV. A number of aspects of virus‐cleaned planting stocks are identified, including reinfection rates, that need investigating before their use is considered as sustainable in developing countries. Popular East African cultivars appear to sustain their long‐term survival by reverting from symptomless infection. The likely biochemistry of this is discussed, and parallels are drawn with other crops. It is concluded that breeding for this attribute will be the best strategy for achieving long‐term control of most sweetpotato viruses.