Diagnosis of light leaf spot (Pyrenopeziza brassicae) on winter oilseed rape (Brassica napus) in the UK

Light leaf spot lesions were generally first observed as light green areas on leaves of UK winter oilseed rape crops in January or February and later became brittle and bleached. Elongated lesions, which were brown with indistinct edges, developed on stems in the spring and summer, when lesions were also observed on flower buds, pedicels and pods. Development of diagnostic white pustules (spore masses of Pyrenopeziza brassicae, which erupt through surfaces of infected tissues) for confirmation of light leaf spot infection on symptomless plants or plants with indistinct or ambiguous symptoms in the autumn, winter or spring was enhanced by incubating plants in polyethylene bags. In experiments with artificially inoculated plants, glasshouse-grown plants exposed in infected crops and plants sampled from crops, white pustules developed at all incubation temperatures from 2^oC to 20^oC on infected leaves of different cultivars. The period of incubation required before the appearance of pustules decreased as the time that had already elapsed since the initial infection increased. The longest periods of incubation were required at the lowest temperatures (2^oC or 5^oC) but leaves senesced and abscised from plants most quickly at the highest temperatures (15^oC or 20^oC), suggesting that the optimal incubation temperature was between 10^oC and 15^oC.     

Effects of light leaf spot (Pyrenopeziza brassicae) on yield of winter oilseed rape (Brassica napus)

The relationship between development of light leaf spot and yield loss in winter oilseed rape was analysed, initially using data from three experiments at sites near Aberdeen in Scotland in the seasons 1991/92, 1992/93 and 1993/94, respectively. Over the three seasons, single-point models relating yield to light leaf spot incidence (% plants with leaves with light leaf spot) at GS 3.3 (flower buds visible) generally accounted for more of the variance than single-point models at earlier or later growth stages. Only in 1992/93, when a severe light leaf spot epidemic developed on leaves early in the season, did the single-point model for disease severity on leaves at GS 3.5/4.0 account for more of the variance than that for disease incidence at GS 3.3. In 1991/92 and 1992/3, when reasonably severe epidemics developed on stems, the single-point model for light leaf spot incidence (stems) at GS 6.3 accounted for as much of the variance. Two-point (disease severity at GS 3.3 and GS 4.0) and AUDPC models (disease incidence/severity) accounted for more of the variance than the single-point model based on disease incidence at GS 3.3 in 1992/93 but not in the other two seasons. Therefore, a simple model using the light leaf spot incidence at GS 3.3 (x) as the explanatory variable was selected as a predictive model to estimate % yield loss (y_r): y_r= 0.32x– 0.57. This model fitted all three data sets from Scotland, When data sets from Rothamsted, Rosemaund and Thurloxton in England were used to test it, this single-point predictive model generally fitted the data well, except when yield loss was clearly not related to occurrence of light leaf spot. However, the regression lines relating observed yield loss to light leaf spot incidence at GS 3.3 often had smaller slopes than the line produce, by the model based on Scottish data.     

Methods for assessment of light leaf spot (Pyrenopeziza brassicae) on winter oilseed rape (Brassica napus) in the UK

Light leaf spot (Pyrenopeziza brassicae) was assessed as % plants with light leaf spot, % leaves with light leaf spot or % leaf area with light leaf spot in winter oilseed rape field experiments done at different sites (Rothamsted, Hertfordshire; Boxworth, Cambridgeshire; near Aberdeen, Scotland), with different cultivars (e.g. Bristol and Capitol), different fungicide treatments, on plants sampled at different dates. Regression analyses on data from these experiments showed that there were consistently good relationships between % leaves with light leaf spot and % plants with light leaf spot for plants sampled during the autumn and winter, until the % plants with light leaf spot approached 100%. The slopes and positions of regression lines were sometimes affected by cultivar, fungicide treatment or sampling date, but not by site. The relationship between % leaf area with light leaf spot (square root-transformed) and % leaves with light leaf spot was less consistent than that between % leaves with light leaf spot and % plants with light leaf spot and was sometimes affected by cultivar, fungicide treatment or sampling date but not by site. The relationship between % leaf area with light leaf spot (square root-transformed) and % plants with light leaf spot was also inconsistent and was sometimes affected by cultivar, fungicide treatment, sampling date and site.     

Effects of previous cropping and fungicide timing on the development of light leaf spot (Pyrenopeziza brassicae), seed yield and quality of winter oilseed rape (Brassica napus)

Light leaf spot, caused by Pyrenopeziza brassicae, was assessed regularly on double-low cultivars of winter oilseed rape during field experiments at Rothamsted in 1990-91 and 1991-92. Previous cropping and fungicide applications differed; seed yield and seed quality were measured at harvest. In each season, both the initial incidence of light leaf spot and the rate of disease increase were greater in oilseed rape crops sown after rape than those sown after cereals. The incidence of diseases caused by Phoma lingam or Alternaria spp. was also greater in second oilseed rape crops. In 1991-92 there was 42% less rainfall between September and March than in 1990-91, and much less light leaf spot developed. However, P. lingam and Alternaria spp. were more common. Only fungicide application schedules including an autumn spray decreased the incidence of light leaf spot on leaves, stems and pods, as indicated by decreased areas under the disease progress curves (AUDPC) and slower rates of disease increase. Summer sprays decreased incidence and severity of light leaf spot on pods only. In 1990-91, all fungicide treatments which included an autumn spray increased seed and oil yields of cv. Capricorn but only the treatment which included autumn, spring and summer sprays increased yields of cv. Falcon. No treatment increased the yields of cv. Capricorn or cv. Falcon in 1991-92. Fungicide applications decreased glucosinolate concentrations in the seed from a crop of cv. Cobra severely infected by P. brassicae in 1990-91, but did not increase yield.     

Identification of QTL involved in field resistance to light leaf spot (Pyrenopeziza brassicae) and blackleg resistance (Leptosphaeria maculans) in winter rapeseed (Brassica napus L.)

 Quantitative trait loci (QTL), involved in the polygenic field resistance of rapeseed (Brassica napus L.) to light leaf spot disease, were mapped using 288 DNA markers on 152 doubled-haploid (DH) lines derived from the cross ‘Darmor-bzh’בYudal’. Over two years (1995 and 1996), the DH population was evaluated for light leaf spot resistance on leaves (L) and stems (S), and for blackleg disease resistance in same field trials. For the L resistance criterion, a total of five and seven QTL were detected in 1995 and in 1996 respectively, accounting for 53% and 57% of the genotypic variation. For the S criterion, three and five QTL were identified in 1995 and in 1996 respectively, explaining 29% and 43% of the genotypic variation. The locations of the QTL detected were quite consistent over the two years (4- and 2-year common QTL for L and S, respectively). Three genomic regions, located on the DY5, DY10 and DY11 groups, were common to the resistance on leaves and stems. In comparison with the QTL for blackleg resistance described by Pilet et al. (1998), two regions on the DY6 and DY10 groups, were associated with the two disease resistances. These ‘multiple disease resistance’ (‘MDR’) QTL may correspond to genes involved in common resistance mechanisms towards the two pathogens or else to clusters of resistance genes. Received: 21 November 1997 / Accepted: 3 March 1998     

Comparative studies of light leafspot (Pyrenopeziza brassicae) epidemics on the growth and yield of winter oilseed rape

Comparisons of epidemics of light leafspot of differing duration and time of initiation were made in two experiments using a single cultivar of Brassica napus. Fungicide was applied before introduction of disease to prevent infection or some time after inoculation to stop further disease development. In the first experiment, substantial reductions in green leaf area and total plant dry-matter were found at flowering when disease was introduced in the autumn or in January. Plant dry weight at maturity was also greatly reduced in these treatments. The detrimental effect of an epidemic initiated in the autumn was avoided to a large extent if fungicide application began in February. Epidemics initiated in March had only small effects on final dry-matter yield. Seed yield was negatively correlated with the length of the epidemic. In a second experiment, early epidemics initiated in the autumn were halted after different time intervals. Commencing fungicide application even as early as December failed to prevent some loss of dry weight at flowering. At maturity, however, dry weight and seed yield were reduced significantly when fungicide application was delayed until February. Failure to control the disease resulted in a 46% loss of seed yield.     

Sugar transport in Coprinus cinereus

Two transport systems for glucose were detected: a high affinity system with a K_m of 27 μM, and a low affinity system with a K_m of 3.3 mM. The high affinity system transported glucose, 2-deoxy-d-glucose (K_m = 26 μM), 3-O-methylglucose (K_m = 19 μM), d-glucosamine (K_m = 652 μM), d-fructose (K_m = 2.3 mM) and l-sorbose (K_m = 2.2 mM). All sugars were accumulated against concentration gradients. The high affinity system was strongly or completely inhibited by N-ethylmaleimide, quercetin, 2,4-dinitrophenol and sodium azide. The system had a distinct pH optimum (7.4) and optimum temperature (45°C). The low affinity system transported glucose, 2-deoxy-d-glucose (K_m = 7.5 mM), and 3-O-methylglucose (K_m = 1.5 mM). Accumulation again occurred against a concentration gradient. The low affinity system was inhibited by N-ethylmaleimide, quercetin and 2,4-dinitrophenol, but not by sodium azide. The rate of uptake by the low affinity system was constant over a wide temperature range (30–50°C) and was not much affected by pH; but as the pH of the medium was altered from 4.5 to 8.9 a co-ordinated increase in affinity for 2-deoxy-d-glucose (from 52.1 mM to 0.3 mM) and decrease in maximum velocity (by a factor of five) occurred. Both uptake systems were present in sporelings germinated in media containing sodium acetate as sole carbon source. Only the low affinity system could initially be demonstrated in glucose-grown tissue, although the high affinity system was restored by starvation in glucose-free medium. The half-time for restoration of high affinity activity was 3.5 min and the process was unaffected by cycloheximide. Addition of glucose to an acetate-grown culture inactivated the high affinity system with a half-life of 5–7.5 s. Addition of cycloheximide to an acetate-grown culture caused decay of the high affinity system with a half-life of 80 min. Regulation is thus thought to depend on modulation of protein activity rather than synthesis, and the kinetics of glucose, 2-deoxy-d-glucose and 3-O-methylglucose uptake would be consistent with there being a single carrier showing negative co-operativity.     

The roles of ascospores and conidia of Pyrenopeziza brassicae in light leaf spot epidemics on winter oilseed rape (Brassica napus) in the UK

Ascospores of Pyrenopeziza brassicae were produced in apothecia (cup‐shaped ascomata) on oilseed rape debris. The conidia, which were morphologically identical to the ascospores, were produced in acervular conidiomata was greater than for lesions caused by ascospores. In June 2000, on the ground under a crop with light on the surface of living oilseed rape tissues. Ascospores were more infective than conidia on oilseed rape leaves. The proportion of lesions caused by conidia located on leaf veins leaf spot, numbers of petioles with apothecia decreased with increasing distance into the crop from the edge of pathways. Air‐borne ascospores of P. brassicae were first collected above debris of oilseed rape affected with light leaf spot on 5 October 1998 and 18 September 1999,12 or 23 days, respectively, after the debris had been exposed outdoors. P. brassicae conidia were first observed on leaves of winter oilseed rape on 6 January 1999 and 15 February 2000, respectively, after plots had been inoculated with debris in November 1998 and October 1999. In 1991/92, numbers of ascospores above a naturally infected crop were small from January to April and increased in June and July. P. brassicae conidia were first observed in February and the percentage plants with leaves, stems or pods with light leaf spot increased greatly in May and June. In 1992/93, in a crop inoculated with debris, numbers of airborne ascospores were small from October to January and increased from April to June. P. brassicae conidia were first observed on leaves in late November and light leaf spot was seen on stems and pods in March and June 1993, respectively.     

Evidence for an intestinal Na+: Sugar transport coupling stoichiometry of 2.0

Membrane potentials maintained by normally-energized intestinal epithelium interfere with an accurate determination of the Na⁺: sugar coupling stoichiometry associated with Na⁺-dependent transport systems. The interference is due to the fact that basal Na⁺ influx is itself a potential-dependent event, and sugar transport induces a membrane depolarization which therefore modifies basal Na⁺ entry. New information obtained under circumstances in which the membrane potential is maintained near 0 indicates that the true coupling stoichiometry is 2:1 rather than the commonly-accepted value of 1:1. A 2:1 stoichiometry means that cellular electrochemical Na⁺ gradients are adequate to account for recently observed 70-fold sugar gradients maintained by these cells under certain conditions.     

A novel methyltransferase from the intracellular pathogen Plasmodiophora brassicae methylates salicylic acid

The obligate biotrophic pathogen Plasmodiophora brassicae causes clubroot disease in Arabidopsis thaliana, which is characterized by large root galls. Salicylic acid (SA) production is a defence response in plants, and its methyl ester is involved in systemic signalling. Plasmodiophora brassicae seems to suppress plant defence reactions, but information on how this is achieved is scarce. Here, we profile the changes in SA metabolism during Arabidopsis clubroot disease. The accumulation of SA and the emission of methylated SA (methyl salicylate, MeSA) were observed in P. brassicae‐infected Arabidopsis 28 days after inoculation. There is evidence that MeSA is transported from infected roots to the upper plant. Analysis of the mutant Atbsmt1, deficient in the methylation of SA, indicated that the Arabidopsis SA methyltransferase was not responsible for alterations in clubroot symptoms. We found that P. brassicae possesses a methyltransferase (PbBSMT) with homology to plant methyltransferases. The PbBSMT gene is maximally transcribed when SA production is highest. By heterologous expression and enzymatic analyses, we showed that PbBSMT can methylate SA, benzoic and anthranilic acids.     

Identification of genes from the obligate intracellular plant pathogen, Plasmodiophora brassicae

Plasmodiophora brassicae is an intracellular pathogen that infects plants in the Brassicaceae family. Although an important pathogen group, information on the genomic makeup of the plasmodiophorids is almost completely lacking. We performed suppression subtractive hybridization (SSH) between RNA from P. brassicae-infected and uninfected Arabidopsis tissue, then screened 232 clones from the resulting SSH library. In addition, we used an oligo-capping procedure to screen 305 full-length cDNA clones from the infected tissue. A total of 76 new P. brassicae gene sequences were identified, the majority of which were extended to full length at the 5' end by the use of RACE amplification. Many of the unisequences were predicted to contain signal peptides for ER translocation. Although we located few sequences in total, these markedly increase available data from the plasmodiophorids, and provide new opportunities to examine plasmodiophorid biology. Our study also points towards the best methods for future plasmodiophorid gene discovery.     

小猿叶甲对寄主植物衰老和缺乏的适应

小猿叶甲Phaedon brassicae Baly是十字花科蔬菜上的一种重要害虫,具有复杂的生活史。测定小猿叶甲取食幼嫩和衰老小白菜叶片后的滞育比率和食料缺乏条件下小猿叶甲成虫的存活时间。研究结果表明,在25℃配合光周期12L∶12D条件下,小猿叶甲取食衰老期小白菜叶片后的滞育比率显著高于取食幼期小白菜叶片后的滞育比率。在25℃、12L∶12D条件下以幼期小白菜叶片饲养小猿叶甲,其成虫羽化后取食0～7d,再分别转入30℃、14L∶10D,不供食的条件下,其结果表明,雌雄成虫存活时间随取食时间延长而延长;羽化成虫取食5d以上,成虫存活时间显著延长,雌雄成虫平均存活时间均在75d以上。最后,讨论小猿叶甲对寄主植物衰老和缺乏的可能适应机制。     

一串红叶斑病的病原菌鉴定

一串红Salvia splendens叶斑病被发现在杭州市农科所栽培花卉的遮阳网大棚中,是由棒孢霉属真菌侵染引起的。在田间,病原菌侵染植株的茎和叶片,引起当年移植苗发生茎腐、落叶、植株枯萎和死亡。通过对病原菌形态学观察以及核糖体DNA ITS序列分析,证明侵染一串红的病原菌为决明生棒孢霉菌Corynespora cassiicola。致病性试验结果表明,一串红是决明生棒孢霉菌的寄主。种子带菌检验结果显示,种子带菌率仅为1%。     

The effect of Brevicoryne brassicae on leaf area, dry matter distribution and amino acids of the Brussels sprout plant

Twenty-six direct or derived criteria were assessed, eight wk after infestation, of the effect of an initial inoculum of 0, 5, 25 or 50 Brevicoryne brassicae on the growth and dry matter distribution of Brussels sprout plants. Twenty-four of these criteria showed significant (P > 0.05) effects of aphid infestation, though increasing aphid infestation above 5/plant had little further effect. Total fresh and dry weight (as well as of the components — leaves, stems and roots) was reduced by 25–730 with infestation, particularly so the root. The number of leaves as well as individual leaf area was reduced, and leaves on infested plants were less efficient assimilators. This could be explained by the drain on assimilates caused by the aphids, the leaf curling and the reduced density of stomata. The latter was also considered to account partly for the increased moisture content of infested plants. There was a general reduction in the concentration of leaf amino acids.     

Sugar transport in leaf discs of Phaseolus coccinius

Kinetic profiles for sucrose, glucose and 3-OMG glucose were determined in leaf discs of Phaseolus coccinius L. (cv. Scarlet). All three sugars exhibited identical uptake kinetics. At sugar concentrations below 25 m M , transport was due to an active, carrier-mediated transport system. A linear component was the dominant mode of uptake at sugar concentrations above 25 m M . Sucrose and glucose carriers were specific for these sugars, since no uptake inhibition was observed from competing sugars. Sucrose was not hydrolyzed by leaf tissue because the label in asymetrically labeled sucrose was not randomized. Furthermore, no label was present in hexose fractions when tissue was incubated with [⁸⁴C]-sucrose. Therefore, [¹⁴C]-sucrose uptake did not reflect hexose uptake.
Both saturable and linear components of uptake contribute significantly to total uptake rates. The former, however, is more important when apoplastic sugar concentrations are low. The molecular nature of the linear component is not well understood but accounts for most of the uptake at high sugar concentrations.     

小猿叶甲成虫越冬和越夏:滞育和休眠同时存在

小猿叶甲Phaedon brassicae Baly是十字花科蔬菜上的一种重要害虫,具有复杂的生活史。2005～2006年在武汉地区对小猿叶甲越冬和越夏虫态及成虫生理状态的调查结果表明,小猿叶甲只以成虫越夏,主要以成虫越冬,但也有少量卵、幼虫和蛹越冬。越夏期间,滞育成虫占70.5%～97.1%。越冬期间,滞育成虫的比率占24.8%～48.2%,随越冬进程,滞育比率上升,但大部分个体处于休眠状态。     

昆诺阿藜链格孢叶斑病病原及其生物学特性

为明确昆诺阿藜链格孢叶斑病病原,在山西省昆诺阿藜种植区采集典型症状的标本分离病原菌,选择代表性菌株LGB-b和LGB-h对其形态学、分子生物学、致病性及生物学特性进行了研究。综合形态学和多基因系统发育(Alt a 1、endoPG 和OPA10-2)分析,确定昆诺阿藜链格孢叶斑病病原为Alternaria alternata。致病性测定发现接种6 d后病斑呈灰绿至黄绿色,表面具有灰棕至灰褐色霉层,周围具黄绿色晕圈,与田间症状基本一致。菌株LGB-b和LGB-h均可侵染昆诺阿藜、藜和台湾藜。菌株LGB-b菌丝生长的最适培养基为V8 蔬菜汁琼脂培养基(V8)、温度为25-30 ℃、水活度≥0.98、pH为6-7;分生孢子萌发的最适水活度≥0.98、pH为6-7。菌株LGB-h菌丝生长的最适培养基为马铃薯胡萝卜琼脂培养基(PCA)、温度为20-25 ℃、水活度≥0.98、pH为6-7;分生孢子萌发的最适水活度≥0.98、pH为7-8。     

浙江番茄灰叶斑病病原菌

近年浙江省杭州市余杭区温室大棚内番茄灰叶斑病发生为害严重,对当地番茄生产造成了较大的经济损失,但该病害的病原菌种类尚未在浙江报道。本文通过对病原菌形态学、分子系统学（rDNA-ITS、gpd和EF-1α基因的PCR扩增、序列分析和系统进化树的构建）和致病性测定相结合的研究,将侵染番茄的病原菌鉴定为番茄匍柄霉菌Stemphylium lycopersici。研究结果对制定该病害的防治决策和有效防治提供了科学理论依据。