Genetic variation among asexual progeny of Phytophthora infestans detected with RAPD and AFLP markers

Genotypic variation among 32 single-zoospore isolates (SZI) of Phytophthora infestans , derived asexually from two hyphal-tip parental isolates (PI-105 and PI-1) of the US-8 genotype, was assessed with 80 random amplified polymorphic DNA (RAPD) primers and 18 amplified fragment length polymorphic DNA (AFLP) primer pairs. In previous investigations, the SZIs from parental isolate PI-105 showed high levels of virulence variability and were differentiated into 14 races, whereas the SZIs from PI-1 showed identical virulence to the parent. The purpose of this investigation was to determine if phenotypic variation observed among SZIs of P. infestans could be detected at the DNA level in these isolates. Polymorphism was detected with 51 RAPD primers and with all 18 AFLP primer pairs in PI-105 SZIs. In SZIs from PI-1, polymorphism was also detected with 25 RAPD primers and 17 AFLP primer pairs. Cluster analysis using the unweighted pair-group method with arithmetic averages (UPGMA) separated the SZIs from parent PI-105 into six virulence groups, 11 RAPD groups and three AFLP groups. Cluster analysis of PI-1 SZIs, which all belong to the same virulence group, differentiated them into four RAPD groups and six AFLP groups. No close correlation among RAPD, AFLP and virulence groups could be established within the two progenies of SZIs. Results of this study suggest that there is a considerable level of inherent genetic variability among SZIs derived asexually from the same parental isolate. The possible mechanisms and implications of this genetic variation are discussed.     

Identification ofTomato Mottle Taino Begomovirus strains in Cuban potato fields

The presence of a begomovirus in potato plants with yellow mottle symptoms was determined for the first time in Cuba. The incidence of typical begomovirus-like symptoms in potato plants in some regions of Havana province (Güira de Melena, San José de las Lajas, Güines and Boyeros) during the growing seasons from 1992 to 1998 was in general low. However, in some cultivars belonging to the National Program for Potato Genetic Improvement, the incidence reached 100%. Yield losses, determined in 1992 and 1994, ranged as high as 19% to 56.33% depending on the cultivar. Characterization of the causal agent was done by light microscopy, host range (graft and mechanical transmission), DNA hybridizations, polymerase chain reaction, and restriction fragment length polymorphism analysis. Nucleotide sequence of the amplified fragments revealed the presence ofTomato mottle Taino virus. The virus was transmittedvia tubers and has been detected in mixed infections withPotato virus X and withPotato leaf roll virus. http://www.phytoparasitica.org posting Oct. 20, 2003. The first two authors contributed equally to this work.     

Further Contributions to the Development of a Differential Set of Pea Cultivars (Pisum sativum) to Investigate the Virulence of Isolates of Aphanomyces euteiches

Common root rot (Aphanomyces euteiches Drechs.) has become a very destructive disease in the French pea crops since 1993. For an accurate investigation of the virulence variability among French A. euteiches populations and between French and foreign populations, a new set of differential pea genotypes was developed. Thirty-three American and European pea lines, displaying different levels of resistance, were screened in a growth chamber against two French isolates. Symptoms (disease severity from 0 to 5, evaluating symptom surface on roots and epicotyl) and percentage of top fresh weight (inoculated/uninoculated top fresh weight ratio) were measured. From this screening 12 relatively resistant lines, from various genetic backgrounds, were identified along with a highly susceptible control. This set of 13 genotypes was inoculated under controlled conditions with 14 isolates from France, Sweden, USA, Canada and New Zealand, to investigate genotype–isolate interactions. Root symptoms were rated (disease severity), and a susceptibility/resistance threshold was established at disease severity = 1. Significant quantitative interactions were observed, and five 'resistance patterns' were identified, leading to a set of six pea genotypes: Baccara (susceptible), Capella, MN313, 902131, 552 and PI180693. Fields trials of this set in 1999 and 2000 gave the same resistance rankings than in growth chamber conditions. This set will allow more accurate assessments of the variability in virulence/aggressiveness of A. euteiches isolates from France and foreign countries, and further investigations of the epidemiological and genetic basis of pea–A. euteiches interactions.     

Modelling Plant Disease Epidemics

An epidemic is the progress of disease in time and space. Each epidemic has a structure whose temporal dynamics and spatial patterns are jointly determined by the pathosystem characteristics and environmental conditions. One of the important objectives in epidemiology is to understand such spatio-temporal dynamics via mathematical and statistical modelling. In this paper, we outline common methodologies that are used to quantify and model spatio-temporal dynamics of plant diseases, with emphasis on developing temporal forecast models and on quantifying spatial patterns. Several examples of epidemiological models in cereal crops are described, including one for Fusarium head blight.     

大丽花常见病害识别及防治

本文对大丽花常见病害症状识别及发病规律进行描述，并提出了综合防治措施。     

Occurrence of Mirafiori Lettuce Virus and Lettuce Big-vein Virus in Relation to Development of Big-vein Symptoms in Lettuce Crops

Big-vein disease (BV) of lettuce has been attributed to infection by Lettuce big-vein virus (LBVV), vectored by the soil fungus Olpidium brassicae. The finding of a second soil-borne virus in lettuce, Mirafiori lettuce virus (MiLV), led to a re-investigation of the role of LBVV in big-vein disease, with evidence emerging that both MiLV and LBVV are vectored by O. brassicae, and that MiLV, not LBVV, is the cause of BV (Lot et al. (2002), Phytopathology 92: 288–293). The two viruses have coat proteins of similar size but have different morphologies and are serologically unrelated. We tested individual lettuce plants in BV-prone fields and protected crops in France and Italy for the presence of the two viruses, using DAS-ELISA and antisera specific for each virus. Both MiLV and LBVV were found at high incidence, often together but sometimes separately. Symptoms were frequently found to be associated with MiLV alone or both viruses, but rarely LBVV alone. However, no absolute correlation emerged, because sometimes MiLV was present in the absence of symptoms, and vice versa. To clarify the situation, individual lettuce plants were examined over a period of time in two further surveys. In surveys of protected crops in France, plants with big-vein were always ELISA-positive for MiLV, or else symptomless plants positive for MiLV were later seen to develop big-vein symptoms. Presence or absence of LBVV appeared to have no effect on symptom development. In surveys of open fields in Italy, all combinations were found: presence of both viruses, apparent absence of both viruses, or presence of each one alone, in plants that developed BV. At the end of the observation period, nearly all plants had BV and contained both viruses.     

Evidence that a Leaf-Disk Test Allows Assessment of Isolate-Specific Resistance in Brassica oleracea Crops Against Downy Mildew (Peronospora parasitica)

A rapid resistance/susceptibility test for Peronospora parasitica (downy mildew) was established by inoculating leaf-disks of four Brassica oleracea accessions. Several conditions were tested: disk disinfection or not, agar medium with or without nutrients and with 50 or 100 ppm of benzimidazole. Using disinfected disks placed on agar (no nutrient and benzimidazole at 50 or 100 ppm), the responses of leaf-disks to four isolates were similar to those obtained using the classical cotyledon test, whereas undesired contaminations occurred in all other conditions. The possible effect of the particular leaf used for obtaining the disks was also studied. In each incompatible interaction tested, disks were resistant whatever the leaf used. In compatible interactions, susceptible phenotypes were observed on disks derived from the six lowest leaves, but disks from upper leaves were resistant. The genetic basis of resistance in a F1 hybrid broccoli was assessed, by testing six isolates on an F2 population derived from this hybrid. The cotyledon test only allows inoculation of two isolates per seedling, whereas many isolates can be tested on each plant by using leaf-disks. The segregation of the resistance to each of the six isolates was analysed: two dominant genes (tightly linked) control resistance to all isolates (one to five isolates; the other to only one isolate).     

节能日光温室中番茄灰霉病发生规律的研究

通过 3年对节能日光温室环境温湿度监测、病菌孢子捕捉和番茄灰霉病发生规律研究明确 ,一般年份 ,灰霉病在番茄叶片上表现为明显的始发期、盛发期和末发期等 3个阶段 ;果实发病后 ,即进入盛发期。温室中病菌的发生与病害呈正相关 ,病菌的高峰期较病害的高峰期提前10～15d。持续的低温、高湿、苗期带病等是引起番茄灰霉病发生的重要因子。提出要从优化设施环境等方面来控制番茄灰霉病的危害。     

引起糖甜菜细菌性叶斑病的萎蔫短小杆菌新致病变种

 1995年在内蒙古临河市新发现了糖甜菜细菌性叶斑病,从病斑所分离的10个细菌菌株经柯赫氏法则验证,均确系该病的病原菌。采用形态观察、表型特征和生理生化特性测定、数值分析、血清学反应、细胞化学成分分析、DNA G+C mol%和DNA-DNA同源性测定进行了鉴定,并与植物病原棒形细菌15个标准菌株进行了比较。该病原菌为革兰氏阳性细菌,不规则短杆状,有一根鞭毛、亚极生或侧生,结合其生理生化特性、细胞化学成分和DNA G+C mol%和DNA-DNA同源性测定结果,认为应属于短小杆菌属(Curtobacterium)的萎蔫短小杆菌(Cur. flaccumfaciens),数值分析也支持这一结论。此外,据血清学反应结果及其对短小杆菌属的其它植物寄主的致病情况,认为该病原菌应是萎蔫短小杆菌种下的一个新的致病变种,定名为Curtobacterium flaccumfaciens pv. beticola pv. nov. Chen et al.,2000(萎蔫短小杆菌糖甜菜致病变种)。     

香蕉束顶病毒(BBTV)侵染对寄主内源激素的影响

 本试验用ELISA方法测定了香蕉感染束顶病毒(BBTV)后植株体内的3种内源激素,赤霉素(GA₃)、玉米素类(iPAs)和脱落酸(ABA)的变化。结果表明,感株的GA₃水平在侵染过程中虽有微弱增加,但含量和增长速度显著低于健株对照;iPAs在接种BBTV第14天后明显下降,并维持较低水平;ABA在BBTV侵染后被大量诱导增加并不断积累,在接种后第35天测定含量最高,为对照的3.34倍。试验还同时检测了BBTV在侵染过程中的运转。用间接ELISA测定的接种叶和顶叶的BBTV含量显示:接种21天后BBTV在接种叶和顶叶中还大量增殖,呈系统性分布。但寄主的症状在接种35天后才逐渐在顶叶表现。以上结果表明:香蕉束顶病的症状表现似乎主要与病株中的内源激素的失调有关,而与BBTV在体内的运转并不直接相关