Detection and quantification ofSpongospora subterranea f. sp.subterranea by PCR in host tissue and naturally infested soils

A polymerase chain reaction (PCR) assay using primers SsF and SsR designed from the internal transcribed spacer (ITS) regions ofSpongospora subterranea f. sp.subterranea was developed for the specific identification and quantification ofS. subterranea. These primers amplified a 434 bp product from DNA ofS. subterranea spore balls, but not from DNA of healthy potato, common scab tuber, and taxonomically related plasmodiophorids. This PCR assay was successfully used for the detection ofS. subterranea in naturally infected symptomatic and asymptomatic potato tubers.Spongospora subterranea in other infected symptomless host plants was detected by PCR. The PCR assay was modified with improved soil DNA extraction methods to detectS. subterranea in soil. The assay was sensitive, and one spore ball per gram of soil could be detected. Following the design of a heterologous competitor DNA template from the sequence of λDNA, a competitive PCR assay for the quantification ofS. subterranea in soil was developed and provided accurate quantification in the range of 1 to 10⁴ spore balls per 0.25 g of soil. In a preliminary survey of naturally infested field soil samples, spore ball concentrations were estimated to vary from ca 0 to 3600 spore balls per 0.25-g soil sample by this competitive PCR assay. The spore ball levels were compared with the powdery scab disease incidence of potatoes in these fields, and a correlationship between spore ball levels and subsequent disease incidence was found. The PCR assays developed in this investigation can be routinely used to detect and quantifyS. subterranea in diseased plant tissue, asymptomatic plant tissue, and infested soil.     

Genetic variation and phylogeny ofSpongospora subterranea f.sp.subterranea based on ribosomal DNA sequence analysis

The nuclear rDNA regions of the two internal transcribed spacers (ITS1 and ITS2) and 5.8S rRNA gene from 52 field isolates ofSpongospora subterranea f.sp.subterranea obtained from the British Isles and North America were polymerase chain reaction-amplified, sequenced, and assessed for genetic variation. Two genetically distinct groups (I and II) were identified based on the ITS sequence diversity among the isolates, representing 34.6% and 65.4% of the isolates, respectively. British Isles isolates occurred in groups I and II, whereas North American isolates belonged only to group II. The British Isles groups ofS. subterranea were associated with particular potato cultivars. The full-length small-subunit rRNA gene ofS. subterranea was sequenced and analyzed by both neighbor-joining and parsimony methods to clarify the taxonomic position of this pathogen. The results of phylogenetic analysis showed thatS. subterranea grouped together with other species of plasmodiphorids, and this group clustered with the phylum Cercozoa, an assemblage of filose and reticulose amoebae and phylogenetically related zooflagellates. The recognition of the existence of different genetic groups withinS. subterranea will be important for the design of plant-breeding programs and in testing for plant resistance.     

Potato powdery scab, caused bySpongospora subterranea f. sp.subterranea, in Costa Rica

Summary Seven samples of potato powdery scab like-lesions were used to determine and confirm the occurrence ofSpongospora subterranea f.sp.subterranea in Costa Rica. Light microscope observations of spore balls and zoosporangia in tomato bait plants identified the organism asS. subterranea. Positive DAS-ELISA test results from all seven samples and PCR specific amplification products from five samples confirmed its identity. The geographical location of the samples source plantations suggests wide distribution of the plant pathogen across Costa Rica main potato growing areas. Different lesion types may indicate host-pathogen or environment-pathogen interactions. The symptoms differences and PCR amplification results among specific primer pairs suggest that some degree of genetic variation among theS. subterranea samples may exist; however, further testing is required.     

Alternative hosts for potato mop-top virus, genus Pomovirus and its vectorSpongospora subterranea f.sp. subterranea

Summary Seventeen weed species common in the potato fields in Denmark were grown in a hydroponic system infested with viruliferous zoospores ofSpongospora subterranea f.sp.subterranea carrying potato mop-top virus (PMTV). The plants were examined for infection with PMTV andS.s.s. using DAS-ELISA and based on visible symptoms. Only two weed species were found to be infected with PMTV,Chenopodium album andSolanum nigrum, whereas 13 became infected withS.s.s. C. album was infected with PMTV by mechanical inoculation were the infection became systemically and the leaves showed necrotic spots.S. nigrum became local infected with PMTV by mechanical inoculation. By inoculation with viruliferousS.s.s. the roots ofS. nigrum became infected but in three weeks PMTV had not spread into the top of the plants.Nicotiana benthamiana was used as a control susceptible toS.s.s. and PMTV.     

Improved diagnosis of powdery scab (Spongospora subterranea f.sp.subterranea) symptoms on potato tubers (Solanum tuberosum L.)

Summary Based on visual inspection, discrimination between common scab (Streptomyces spp.) and powdery scab (Spongospora subterranea) can be difficult. Inspections are performed on unwashed samples, incidentally supported by microscopic examination. During 1994–1996 surveys were performed in The Netherlands on tubers with symptoms resembling common scab. Under microscopic assessment nearly all samples showed the presence of structures resembling cystosori (sporeballs) ofS. subterranea. At that time confirmation using alternative techniques was not possible. In 2003 research was undertaken to clarify the situation with respect to scab on potato tubers in The Netherlands. One hundred and eighteen scab samples were extensively tested forS. subterranea. All samples were digitally photographed, microscopically examinated and tested with real-time PCR and DAS-ELISA. Use of these modern methods resulted in a clear picture of symptoms that can be attributed toS. subterranea. A lot of scab samples with structures resembling cystosori could not be confirmed as contaminated withS. subterranea.     

Experimental host range and natural reservoir of sweet potato leaf curl virus in the United States

Sweet potato leaf curl virus (SPLCV), a sweet potato whitefly (Bemisia tabaci) transmitted begomovirus, causes serious yield losses to many sweet potato cultivars. Using experimental whitefly transmissions in a greenhouse (choice tests) and in a growth chamber (no-choice tests), we evaluated 111 plant species in 30 families to determine the host range of SPLCV. The host range was limited to plants in the genus Ipomoea within the family Convolvulaceae. In total, 38 of 45 Ipomoea species tested were susceptible to SPLCV infection. Surveys were conducted during the 2007-2009 sweet potato growing seasons in Mississippi and South Carolina to evaluate morning glory species as potential reservoir hosts for SPLCV. In the sweet potato experimental fields and surrounding areas, a large proportion of volunteer sweet potatoes, as well as a high percentage of annual and perennial morning glories tested positive for SPLCV. Understanding the host range and potential virus reservoir host plants will ultimately help in the development of an effective disease management strategy that is based on the consideration of agroecological factors.     

2种激发子对西瓜枯萎病的诱抗作用

分别测定了来源于西瓜枯萎病菌和香蕉枯萎病菌的细胞壁来源的水溶性、热稳定激发予对西瓜枯萎病的诱抗作用。结果表明：2种来源的激发子均有明显的诱导西瓜苗细胞壁木质化的作用，但来源于香蕉枯萎病菌的激发子对木质化的诱导活性比来源于西瓜枯萎病菌的要高；西瓜苗分别经2种激发子处理后，体内过氧化物酶、多酚氧化酶和苯丙氨酸解氮酶的活性均有异常的增加，但来源于西瓜枯萎病菌的激发子对这3种防御酶的诱导速度比来源于香蕉枯萎病菌的要快；经激发子诱导处理后瓜苗对枯萎病菌的抗性有了明显的提高，但来源于西瓜枯萎病菌的激发予的诱抗效果相对较好。     

香蕉枯萎病菌粗毒素的毒性及其模型

以香蕉组织培养苗为受试植物,蕉苗受毒素作用后的病情指数为指标,测定香蕉枯萎病菌(Fusarium oxysporum f.sp.cubense)粗毒素对蕉苗的毒性。结果表明,毒素处理72,96,120和144h引致香蕉苗受害程度(以病情指数表示)达90的剂量(TD90)的质量浓度分别为2057.20,1245.49,549.54,380.19μgmL,蕉苗的受害程度存在着随处理时间延长和处理剂量的增加而加重的趋势。根据毒素对蕉苗的毒性测定结果,建立了蕉苗受毒素作用的“时间-剂量-受害程度”模型。根据模型的分析结果认为,粗毒素引致蕉苗受害的有效时间为96~120h,有效剂量质量浓度为260.0~130.0μgmL。      

美国玉米种质的利用与改良

新种质的普遍缺乏和遗传基础狭窄,已经成为玉米育种者所面临的难题。美国作为世界上玉米播种面积及总产量最高的国家,在玉米种质的理论及应用上都取得了突出的成果。本文回顾、总结了美国在种质利用方面的历史和现状,分析了杂种优势理论的发展和杂种优势模式的应用,以及通过群体改良、外来种质导入手段保持遗传多样性,改良、拓宽种质基础的理论发展及其实践。     

海南省香蕉枯萎菌nit突变体的筛选及鉴定

将23个有致病力的香蕉枯萎病菌海南菌株及属于1,4号生理小种菌株Foc-18,Foc-38分别在含氯酸盐的KPS培养基上培养,23个菌株共产生142个抗氯酸盐突变体,诱发获得98个不能利用硝酸盐的营养缺陷突变体。将获得的nit突变株作营养体亲和性配对试验,可将23个菌株分为4个亲和群,其中VCG1包括11个菌株,且全为海南分离的1号生理小种,VCG2则由10个海南4号小种菌株组成,广东1号小种Foc-18,4号小种Foc-38分别属于VCG3,VCG4。     

亚麻枯萎病病原菌生理小种研究进展

生理小种是亚麻枯萎病病原菌致病性分化的标志之一,迄今为止,全国已有十余年未对亚麻枯萎病病原菌生理小种进行系统的研究.因此,本文在阐述亚麻枯萎病发生、流行及防治的基础上,重点概括了其病原菌生理小种的分化原因、鉴定方法、研究现状及作物抗病育种相关内容,同时提出了相应的可行性建议,为后续学者深入研究亚麻枯萎病的发生、流行规律以及病害防治等提供相关依据.     

香蕉枯萎菌原生质体形成与再生条件

通过对菌丝菌龄、酶液浓度、酶解时间、酶解温度、渗透压稳定剂的种类和浓度等因素的实验研究,得到了一种制备香蕉枯萎菌(Fusarium oxysporum f.sp.cubense,FOC)原生质体的有效方法,并在固体再生培养基上实现了原生质体的再生,再生率为22.5%。     

Resistance toSpongospora subterranea in tuber-bearing and non tuber-bearingSolanum spp.

Summary The incidence of infection withSpongospora subterranea was studied in the non tuber-bearingSolanum brevidens andS. etuberosum and the tuber-bearingS. acaule, S. sucrense andS. tuberosum cvs Olympia and Pito. Shoot cuttings were grown in soil naturally infested withS. subterranea, or the roots were inoculated with a zoospore suspension. Logit models were used to analyse the data. The incidence ofS. subterranea was higher in plants inoculated with zoospore suspension than in those grown in infested soil (odds ratio (OR) 10.65). Ageing the inoculum reduced the incidence of infection in the plants (OR 0.30) without altering the interspecific differences. The ORs of infection (compared to cv. Olympia) were 0.07, 0.29, 0.60 and 2.88 forS. acaule, S. sucrense, S. brevidens andS. etuberosum. OnlyS. acaule was significantly more resistant to infection than cv. Olympia. No infection was detected in cv. Pito.     

我国燕麦秆锈菌生理小种与毒力分析

为了解当前我国燕麦秆锈菌生理小种及其毒力现状,利用12个北美燕麦秆锈菌单基因鉴别寄主,对2013年采集的7份和2012保存的4份燕麦秆锈病标样进行小种与毒力鉴定,并用Pg a-code字母命名系统命名小种,以16个单基因系测定毒力频率。结果表明,从11份燕麦秆锈菌标样中分离成活了26个单孢子堆菌株,鉴定出TKR、TJM和TKM三个生理小种,出现频率分别为61.6%、30.8%和7.6%。26个菌株对Pg 6和Pg 15无毒力,对Pg 10、Pg 13、Pg a和Rodney 0的毒力频率分别为69.2%、61.6%、61.6%和84.6%,对Pg 1、Pg 2、Pg 3、Pg 4、Pg 8、Pg 9、Pg 12、Pg 16和Marvellous毒力频率高达100%。说明当前我国燕麦秆锈菌优势小种为TKR,次优势小种为TKM;Pg 6和Pg 15为有效抗病基因,Pg 1、Pg 2、Pg 3、Pg 4、Pg 8、Pg 9、Pg 12、Pg 16和Marvellous则为无效抗病基因,Pg 10、Pg 13、Pg a和Rodney 0对我国燕麦秆锈菌具有一定的区分作用。     

尖孢镰刀菌胡麻专化型(Fusarium oxysporum f . sp. lini) ISSR标记聚类分析

结合形态学特征和分子生物学方法对我国尖孢镰刀菌胡麻专化型菌株进行鉴定、聚类分析和遗传变异分析。结合传统分类方法和ITS序列分析,确定6省区96株菌株为尖孢镰刀菌。采用ISSR（简单重复序列区间）分子标记技术对这96株菌株进行分析,12条引物共扩增出800个条带,多态性条带数为797条,多态性为99.62%;在相似性系数为0.88处,96株供试菌株被分为5个类群。聚类结果表明,胡麻枯萎病菌种内存在明显的多态性,且ISSR类群与地理来源存在相关性。