黑胫病菌（Leptosphaeria biglobosa）在油菜叶片和茎中的侵染过程观察

为明确黑胫病菌（Leptosphaeria biglobosa）在甘蓝型油菜叶片和茎中的侵染及扩展过程，利用绿色荧光蛋 白（GFP）标记的黑胫病菌株接菌油菜叶片，利用激光共聚焦显微镜观察菌株在油菜叶片和茎中的侵染过程。结果 表明，接种油菜叶片7 h后，分生孢子萌发并长出芽管；17 h后，芽管侵入气孔；24 h后，分生孢子全部萌发；36 h后萌 发的芽管形成菌丝；120 h后，菌丝在叶片表皮细胞间隙蔓延，并侵入叶肉细胞。13 d后，菌丝侵入茎部皮层组织； 15 d后，菌丝在皮层细胞间隙蔓延，并侵染至茎表皮；21 d后，菌丝侵染至维管组织；23 d后，菌丝侵染至茎韧皮部； 25 d后，茎导管被侵染，并向木质部扩展。本研究发现的L. biglobosa 在油菜叶片和茎中的侵染过程，可为油菜与黑 胫病菌互作的研究、黑胫病致病机理及防治提供参考。     

油菜黑胫病菌T-DNA插入诱变因素优化及突变体筛选

油菜黑胫病是由Leptosphaeria biglobosa引起的一种真菌病害。这种病害在我国油菜产区广泛发生，并造成一定的经济损失。为通过获取突变体来研究L. biglobosa的生态适应性机制及致病机制，本文优化了影响农杆菌介导转化（ATMT）油菜黑胫病菌L. biglobosa菌株Lb731的因素，评估转化子质量，并筛选相关突变体。结果明确了农杆菌介导转化菌株Lb731的最佳因素：潮霉素B浓度为50 μg/mL，转化受体（分生孢子）培养时间为15 d (20℃)，浓度为2 × 10^7~8孢子/mL，农杆菌-受体共培养温度为25℃，共培养时间为72 h。在最适条件下的转化效率达到80个转化子/百万分生孢子。T-DNA插入基因组的频率为100%，单拷贝插入频率为72.7%，转化子抗潮霉素性状能稳定遗传。从2136个转化子中获得了11个菌丝生长减缓突变体，7个色素产生缺陷突变体和14个分生孢子产生缺陷突变体，并从这些突变体中鉴定出7个致病力丧失突变体。采用hiTAIL-PCR技术，从3个突变体中获得了T-DNA插入位点侧翼序列。上述结果为深入研究L. biglobosa的生态适应性机制及致病机制提供了材料和线索。     

Further evidence on the effects of gene H,which confers cane hairyness,on resistance to raspberry diseases

Summary Gene H determines cane hairyness in raspberries and was reported to facilitate escape from infection by Botrytis cinerea and Didymella applanata. However, experiments with mycelial inoculations of wounded canes showed that it conferred a form of resistance which was evident for a long period after the establishment of infections by these pathogens. The gene also contributed to the resistance of fruit to B. cinerea and of canes to Leptosphaeria coniothyrium. Alternative hypotheses to explain these observations are discussed: two based upon its possible genetic linkage with a gene or genes for resistance and one based upon its possible pleiotropic effects.     

Selection for resistance to Septoria nodorum in wheat

Summary A population of 572 F₂ derived F₃ lines from six crosses were used to estimate parameters relevant to selection for resistance to Septoria nodorum of wheat. Lines were grown in disease free (fungicide sprayed) and inoculated microplots in 2 replications of a split-plot design in a single environment in 1977. Average yield reduction due to disease was approximately 50%; this was associated with an average septoria score of 50% on the flag leaf, an average septoria score of 42% on the head, and a reduction of 37% in seed weight. Low S. nodorum scores were correlated with late heading date, tall plant height, high grain yield, and high seed weight in diseased plots, and high seed weight % (seed weight in diseased plots expressed as a percentage of seed weight in fungicide sprayed plots).Restricted selection indexes were used to study the relative contributions of disease escape, true resistance, and tolerance to variability in grain yield in diseased plots, seed weight in diseased plots, and seed weight %. True resistance appeared to be the most important factor causing variation in grain yield in diseased plots and seed weight %. Tolerance and escape seemed to be more important for seed weight in diseased plots.Heritabilities of S. nodorum scores on the flag leaf and head were 63% and 52%, respectively. Leaf and head scores could be used most effectively as selection criteria to upgrade resistance in a population before harvest.Selection for high seed weight % slightly reduced yields in disease free plots, although yield in diseased plots and seed weight in diseased plots were increased. However, selection for increased yield or increased seed weight in diseased plots improved yield in disease free plots. It is suggested that direct selection for yield or seed weight in diseased plots is likely to achieve more desirable goals than selection for seed weight %.     

The Inheritance of Resistance to Septoria Glume Blotch II. The Wild Wheat Species Aegilops speltoides

The inheritance of resistance in the wild wheat species Ae. speltoides L. to Septoria glume blotch (Septoria nodorum Berk.) was investigated. It was characterized by two parameters measured on detached leaves, namely lesion size (LS) and length of latent period (LP), and by the severity of the disease under field conditions (DS). The genetic analysis was based on the F₁ and F₂ generations of a 4 × 4 diallel cross between four Ae. speltoides accessions showing differential resistance to the pathogen. The three parameters of resistance were highly correlated. Considerable heterosis for resistance was found for each of the parameters in most of the diallel combinations. The estimates of broad-sense heritability were moderately high (0.50 — LS, 0.60 — LP, 0.52 — DS), while the estimates of narrow-sense heritability were low (0.16 for LS, 0.20 for LP, 0.25 for DS). There were no indications for genie interaction. It is suggested that the resistance is controlled by at least two genes with over-dominance of the alleles for higher resistance. Highly resistant Ae. speltoides accessions are a potential source of germplasm for improving the resistance of cultivated wheats to Septoria glume blotch. The possibility of using dominant alleles for resistance in hybrid cultivars is discussed.     

Effect of seeders and tillage equipment on vertical distribution of oilseed rape stubble

When the spreading of a disease depends on the proportion of infected residues remaining at soil surface it is of crucial importance to analyse the effects of tillage practices on the vertical distribution of stubble. This is the case with phoma stem canker (blackleg), whose epidemics are initiated in autumn, by air-borne ascospores released from stubble located at the soil surface. We compared initial vertical distribution of oilseed rape residues to those observed after sowing and various tillage operations (rotary harrowing, stubble disking, chiselling and mouldboard ploughing). Almost 20% of the initially buried residue was brought back to soil surface with seeding. Rotary harrow brought 40% of the residue buried in the 0–10 cm layer up to the surface and left unburied about 70% of surface residue. Stubble disking appeared to be more efficient for residue burial than chiselling. Mouldboard plough was the only tool that buried all residues. A simple model was developed that predicted burial and return to the soil surface of potentially infected residues as a function of tillage practices used after harvest. Simulation of different tillage sequences showed that the order in which tools were used also affected location of residues. Our results highlighted the importance of tillage in the cultural control of phoma stem canker and will contribute to the definition of integrated pest management strategies for oilseed rape.     

Current progress in studying blackleg disease (Leptosphaeria maculans and L. biglobosa) of canola in Iran: Where do we stand now?

Blackleg, caused by a complex of Leptosphaeria species (L. maculans and L. biglobosa), is a fungal disease on Brassica species, especially important in canola (Brassica napus). Since the first report of L. biglobosa in Iran in 2007 and L. maculans in 2008, both species are now of major importance in Iran affecting 10 provinces and 30 regions, with a higher prevalence in the northern provinces of Mazandaran and Golestan. Despite the rapid progression of the disease and the emergence of new Leptosphaeria races in Iran, the research into this pathogen has not progressed at the same rate and is limited to phenotypic characterization studies, pathogenicity research, and to a lesser extent, disease management research. Given the rapid increase in canola cultivation in Iran and changes in the genetic diversity of the pathogen populations, it is likely that blackleg disease will increasingly become a severe threat to Iran’s canola production. Therefore, systematic and prospective studies, along with fundamental research on the pathogen's biology, epidemiology, and genetic diversity, would provide critical information for the development of disease management strategies. Here, we review the research that has been carried out to date on blackleg disease in Iran and describe the extent of progress towards disease control, especially in disease-prone regions.     

Effects of temperature on ascospore germination and penetration of oilseed rape (Brassica napus) leaves by A- or B-group Leptosphaeria maculans (phoma stem canker)

Ascospores of both A-group and B-group Leptosphaeria maculans germinated at temperatures from 5 to 20°C on leaves of oilseed rape. Germination of ascospores of both groups started 2 h after inoculation and percentage germination reached its maximum about 14 h after inoculation at all temperatures. Both the percentage of A-/B-group ascospores that had germinated after 24 h incubation and germ tube length increased with increasing temperature from 5 to 20°C. Germ tubes from B-group ascospores were longer than those from A-group ascospores at all temperatures, with the greatest difference at 20°C. Hyphae from ascospores of both groups penetrated the leaves predominantly through stomata, at temperatures from 5 to 20°C. A-group ascospores produced highly branched hyphae that grew tortuously, whereas B-group ascospores produced long, straight hyphae. The percentage of germinated ascospores that penetrated stomata increased with increasing temperature from 5 to 20°C and was greater for A-group than for B-group L. maculans after 40 h incubation.     

A method to test wheat leaves for their reactions to inoculation with Septoria species

The construction and application is described of a polystyrol humidity box in which wheat leaves, while continuing to function as parts of living plants, can be tested for their reactions toSeptoria spp. in an atmosphere nearly saturated with water, as is required for successful infection. The method is simple, accurate ans inexpensive.Samenvatting Voor dit doel is een z.g. vochtdoos geconstrueerd (Fig. 1A). Het te toetsen blad wordt daar doorheen geleid en in de doos geïnoculeerd met een druppel conidiën-suspensie van de schimmel. Onder het blad staat wat water in de doos (Fig. 1B). Na afsluiting ontstaat in de doos de hoge luchtvochtigheid van bijna 100% die nodig is voor de infectie. Dit wordt op deze wijze eenvoudig en goedkoop gerealiseerd. De bladeren die zo worden getoetst blijven onbeschadigd functioneren aan de plant. Na enkele dagen kan de doos worden geopend en kan de symptoom-ontwikkeling worden afgewacht en gevolgd (Fig. 3A, 3B en 4). De methode leent zich voor nauwkeurig werk en vereist zeer weinig infectiemateriaal. De Technische en Fysische Dienst voor de Landbouw (TFDL), Wageningen, ontwierp en construeerde een statief voor het gebruik van de vochtdozen in serie (Fig. 2).     

Differentiating A and B groups of Leptosphaeria maculans, causal agent of stem canker (blackleg) of oilseed rape

Stem canker or blackleg of brassicas, caused by Leptosphaeria maculans , is one of the most damaging diseases of winter oilseed rape in the UK. Airborne ascospores, released in autumn and winter, initiate leaf infections which may lead to colonization of the petiole and, later in the season, formation of stem lesions and cankers. Although isolates of the pathogen differ in ability to cause damaging stem cankers, this is not readily apparent from leaf spotting or stem lesion symptoms. However, several cultural, biochemical and genetic characteristics appear to be associated with the ability to form damaging stem cankers and isolates can be assigned to one of two groups, termed A and B, on the basis of differences in these characteristics. To investigate the relationship between leaf spotting symptoms and subsequent stem canker formation, and to improve understanding of the epidemiology of this pathogen, it is desirable to differentiate between the stem canker forming A group and the less damaging B group of L. maculans . Characterization of isolate type is also important in seed testing and crop breeding programs, particularly in countries such as Canada and Poland where the A type is not ubiquitous. This article reviews methods, including plant assays, assessments of growth characteristics in vitro , isozyme analyses, secondary metabolite profiling, serology, and nucleic acid analyses, that can be used to differentiate the A and B groups.