Life-history ofNeurospora intermedia in a sugar cane field

The life-history ofNeurospora in nature has remained largely unknown. The present study attempts to remedy this. The following conclusions are based on observation ofNeurospora on fire-scorched sugar cane in agricultural fields, and reconstruction experiments using a colour mutant to inoculate sugar cane burned in the laboratory. The fungus persists in soil as heat- resistant dormant ascospores. These are activated by a chemical(s) released into soil from the burnt substrate. The chief diffusible activator of ascospores is furfural and the germinating ascospores infect the scorched substrate. An invasive mycelium grows progressively upwards inside the juicy sugar cane and produces copious macroconidia externally through fire- induced openings formed in the plant tissue, or by the mechanical rupturing of the plant epidermal tissue by the mass of mycelium. The loose conidia are dispersed by wind and/or foraged by microfauna. It is suggested that the constant production of macroconidia, and their ready dispersal, serve a physiological role: to drain the substrate of minerals and soluble sugars, thereby creating nutritional conditions which stimulate sexual reproduction by the fungus. Sexual reproduction in the sugar- depleted cellulosic substrate occurs after macroconidiation has ceased totally and is favoured by the humid conditions prevailing during the monsoon rains. Profuse micro-conidiophores and protoperithecia are produced simultaneously in the pockets below the loosened epidermal tissue. Presumably protoperithecia are fertilized by microconidia which are possibly transmitted by nematodes active in the dead plant tissue. Mature perithecia release ascospores in situ which are passively liberated in the soil by the disintegration of the plant material and are, apparently, distributed by rain or irrigation water.     

印度块菌子囊果内部解剖结构的扫描电镜观察

利用扫描电镜对成熟印度块菌子囊果的内部结构进行了观察,以系统揭示其子囊果内部组织特征,为块菌属的分类以及块菌属真菌子囊果的生理研究奠定基础。观察结果进一步证明,成熟印度块菌子囊果横切面上的白色迷宫状脉络是由不育的侧丝构成,而暗脉则是被侧丝缠绕并包裹着的可育的菌丝组织,即产孢组织,这些白色脉络和暗脉就构成了印度块菌子囊果横切面上迷宫状的纹脉;产孢组织中,可观察到正在发育的大大小小的子囊被缠绕在一起的大量产囊丝与侧丝包裹着,形成密密麻麻微小的类似蜂巢状结构;子囊孢子游离于子囊中,成熟子囊孢子表面有刺状纹饰,刺的顶端有小弯钩。单个子囊内含的子囊孢子大小与其内含的子囊孢子数目有关,子囊内所含的子囊孢子越多,子囊孢子就越小。     

Enhanced pathogenicity of Leptosphaeria maculans Pycnidiospores from paired co-inoculation of Brassica napus cotyledons with ascospores

BACKGROUND AND AIMS: Blackleg, caused by Leptosphaeria maculans, is a major disease of oilseed rape (Brassica napus) worldwide, including Australia. In most cases, the severity of the disease in the field is related to infections caused by airborne ascospores. In contrast, pycnidiospores originating from leaf and stem lesions and stubble are widely assumed to play only a relatively minor role in the epidemiology of blackleg. It is not clear whether, under certain conditions, pycnidiospores can cause severe disease in the field. The aim of the work reported was to determine if the pathogenicity of pycnidiospores is enhanced by paired co-inoculation of B. napus cotyledons with ascospores. METHODS: Three investigations were carried out under controlled-environment conditions using various L. maculans isolates and B. napus cultivars with different levels of host resistance to blackleg. KEY RESULTS: In all three experiments, co-inoculation with ascospores increased the ability of pycnidiospores to cause more disease on B. napus than when inoculations consisted of pycnidiospores alone. This effect was significantly influenced by the host resistance of the cultivar, but overall was independent of the L. maculans isolate used in the different experiments. This effect was also independent of timing of inoculation with the ascospores, with increased disease from pycnidiospores occurring on the cotyledon of the seedling in situations where inoculations with ascospores were carried out 0, 1 or 2 d after pycnidiospore inoculation. This enhanced pathogenicity of pycnidiospores was evident even when low concentrations of pycnidiospores were applied to the other cotyledon of the same seedling. CONCLUSIONS: These results may explain continuing severe blackleg disease cycles throughout the cropping season even when ascospore fallout was low or constrained only to a brief period or phase of the cropping season, and suggest that disease epidemics may be polycyclic rather than monocyclic.     

Formation and Development of Pseudothecia of Venturia nashicola

Conidia are believed to be the main source of primary inoculum for pear scab, caused by Venturia nashicola, in northern China. Experiments were conducted to investigate the development and potential role of V. nashicola ascospores in northern China. Leaves with pear scab lesions were collected from commercial orchards in November 2003 and 2004 to monitor pseudothecia formation under various environments. Pseudothecium production was shown to occur readily in northern China. The key requirement for pseudothecium production is the occurrence of rain during the winter and early spring, although the exact timing of these rain events appeared not to affect their development. Excess water may lead to the accelerated leaf decay and hence lead to production of fewer pseudothecia. More than 80% scabbed leaves, placed in a pear orchard, produced pseudothecia. Leaves with only non‐sporulating scab lesions in autumn were also able to produce a large number of pseudothecia. Both airborne ascospores and conidia of V. nashicola were caught in a pear orchard. Most ascospores were released by late‐May, a month after pear blossom. These results suggest that ascospores may play an important role in the early stage of pear scab epidemics in spring in northern China.     

昆明地区印度块菌子囊果的解剖结构研究

通过印度块菌子囊果石蜡切片的显微观察,对印度块菌子囊果包被及子实层的显微结构、子囊及子囊孢子的发育过程进行研究.结果表明:(1)印度块菌子囊果是由包被和子实层构成,子囊果的表面密被大量大小不一的疣状突起,包被由外皮层和内皮层构成;子实层由封闭的、大小不一的产孢组织构成,在产孢组织中包含有侧丝、产囊丝、子囊和子囊内的子囊孢子;(2)成熟印度块菌子囊果横切面上有明暗相间的迷宫状纹脉;(3)子囊卵形,由产囊丝顶端的细胞发育而来;(4)成熟子囊孢子红褐色,椭圆形至近球形不等,子囊孢子双层壁,外壁密布有刺状纹饰.子囊孢子在子囊内的发育过程中常出现败育现象,每个成熟子囊中含有1～5个子囊孢子(常见4个).     

Symptoms and Prepenetration Events Associated with the Infection of Common Bean by the Anamorph and Teleomorph of Glomerella cingulata f.sp. phaseoli

Glomerella cingulata f.sp. phaseoli and Colletotrichum lindemuthianum are the teleomorph and anamorph, respectively, of the pathogen causing anthracnose in common bean. The mechanisms relating to the sexual reproduction of this plant pathogen are still unclear, as are the infection structures involved and the symptoms produced. In the present study, bean plants were inoculated with ascospores and conidia, and the events taking place within the following 120 h were investigated using light microscopy and scanning electron microscopy. The symptoms exhibited by plants inoculated with the ascospores were milder than in those inoculated with conidia. Microscopy revealed that most of ascospores produced germ tubes and appressoria at an early stage (24 h after inoculation). From 48 h onwards, the formation of hyphae and the production of germ tubes and appressoria were great. In contrast, infections originating from conidia developed more slowly, and at 24 and 48 h, many non‐germinated conidia were present, whereas only few conidia developed germ tubes and appressoria. Ascospore germination and appressorium formation were similar on both resistant and susceptible cultivars. Hence, the symptoms and the temporal sequence of events associated with the infection of bean plants by the two fungal forms differed, although the structures produced were similar. This is the fist report comparing symptoms and prepenetration events between anamorph and teleomorph of G. cingulata f.sp. phaseoli in common bean.     

湖南地区茯砖茶中金花菌的分离鉴定

对湖南地区茯砖茶上＂金花菌＂的菌落特征、形态特征进行了研究,并运用扫描电镜观察了其子囊孢子和分生孢子的发育过程,鉴定其为冠突散囊菌（Eurotium cristatum）。     

印度东北部白垩纪-第三纪Xylariaceae化石孢子

在印度东北部白垩纪-第三纪沉积中发现了各种类型单沟无隔壁的Xylariaceae化石孢子,例如:Hypoxylonites,Spirotremesporites.Hypaxylonites等共计15种,其中6新种,它们是Hypoxylonites bhubanensis,H.elsikii,H.minimus,H.neogenicus,H.subrotundus和H.thindii,Hypoxylonites在本地区一般出现于晚白垩世地层中,占化石微体植物群总数的3%-4%,而在始新世它们的含量上升到植物群总数的20%-25%,从中新世直到早上新世这类孢子类型最多,含量最高,达植物群总数的40%-45%,到晚上新世含量突然下降,而Spirotremesporites在白垩纪未发现,到晚始新世时才出现,仅占孢粉总数的5%-6%,中新世时达25%-30%,到上新世末衰落,该属含5新种,Spirotremespores ellipticus,S.longilentus,S.miocenicus,S.reniformis,S.tertiarus.考虑到和这些种共生的Xylaria,Hypoxylon,Rosellinia及Daldina的孢子以及它们的地层分布,这类孢子的演化历史可能是受沉积盆地附近的古气候控制的.     

Genetic Variability of Hymenoscyphus pseudoalbidus on Ash Leaf Rachises in Leaf Litter of Forest Stands in Poland

Two hundred and thirty cultures of Hymenoscyphus pseudoalbidus were obtained from ascospores created in apothecia on the previous years' ash leaf rachises in the stand floor. Fruiting bodies of the pathogen were collected in four regions of Poland differing by geographical location, the altitude above sea level and climatic conditions. Isolates were identified based on the sequences of ribosomal DNA (ITS1‐5.8S‐ITS2) and the calmodulin gene. Only the presence of H. pseudoalbidus was identified in the decaying ash stands in Poland; morphologically similar, saprotrophic species of H. albidus was absent. Intrapopulation and interpopulation genetic variability of isolates was determined based on 84 RAMS markers obtained using four primers. Genetic variability of the fungus populations, measured by the Dice coefficient of genetic similarity and the Shannon coefficient of genetic diversity, decreased along with a decrease in the location of isolate collection area above sea level. A significant dependency was shown between intrapopulation genetic variability of isolates and altitude of regions above sea level. The Mantel test excluded existence of dependence between geographical and genetic distance among populations (r = ?0.038, P = 0.55). A significant correlation was found between the genetic distances of individuals within populations and locations above sea level. Based on PCA and geographical location of populations, it was shown that populations create four distinct groups. amova showed that a majority of total genetic variability (65.80%) constitutes intrapopulation variability. Variability between populations was high (28.7%), and individual regions had a smallest influence (5.5%) on the level of total variability.     

Temperature and leaf wetness duration affect phenotypic expression of Rlm6-mediated resistance to Leptosphaeria maculans in Brassica napus

Near-isogenic Brassica napus lines carrying/lacking resistance gene Rlm6 were used to investigate the effects of temperature and leaf wetness duration on phenotypic expression of Rlm6-mediated resistance. Leaves were inoculated with ascospores or conidia of Leptosphaeria maculans carrying the effector gene AvrLm6. Incubation period to the onset of lesion development, number of lesions and lesion diameter were assessed. Symptomless growth of L. maculans from leaf lesions to stems was investigated using a green fluorescent protein (GFP) expressing isolate carrying AvrLm6. L. maculans produced large grey lesions on Darmor (lacking Rlm6) at 5-25 degrees C and DarmorMX (carrying Rlm6) at 25 degrees C, but small dark spots and 'green islands' on DarmorMX at 5-20 degrees C. With increasing temperature/wetness duration, numbers of lesions/spots generally increased. GFP-expressing L. maculans grew from leaf lesions down leaf petioles to stems on DarmorMX at 25 degrees C but not at 15 degrees C. We conclude that temperature and leaf wetness duration affect the phenotypic expression of Rlm6-mediated resistance in leaves and subsequent L. maculans spread down petioles to produce stem cankers.