Ultrastructure of the host-pathogen interface in daylily leaves infected by the rust fungus Puccinia hemerocallidis

Summary. Transmission electron microscopy was used to examine details of the host–pathogen interface in daylily leaf cells infected by the rust fungus Puccinia hemerocallidis. Samples were prepared for study by high-pressure freezing followed by freeze substitution. The outstanding preservation of ultrastructural details afforded by this fixation protocol greatly facilitated the study of this host–pathogen interface. The extrahaustorial membrane that separated each dikaryotic haustorium from the cytoplasm of its host cell was especially well preserved and appeared almost completely smooth in profile. Large aggregations of tubular cytoplasmic elements were present near haustoria in infected host cells. Many of these tubular elements were found to be continuous with the extrahaustorial membrane and conspicuous electron-dense deposits present in the extrahaustorial matrix extended into these elements. The use of gold-conjugated wheat germ agglutinin for labeling of chitin revealed that these deposits were not part of the haustorial wall. Portions of many of the tubular elements associated with haustoria were conspicuously beaded in appearance. Some tubular elements were found to be continuous with flattened cisternae that in turn bore short beaded chains. Distinctive tubular-vesicular complexes previously reported only in cryofixed rust haustoria also were found in the haustoria of P. hemerocallidis. Received July 6, 2001 Accepted October 3, 2001     

Effect of temperature, light and host on prepenetration development of Puccinia graminis avenae and Puccinia coronata avenae

The influence of temperature and light on prepenetration development of single and mixed isolates of Puccinia graminis avenae and Puccinia coronata avenae was studied on 0–2% water agar and on leaves of three oat cultivars and on three non-cultivated species of Avena. Germination of uredospores of P. graminis avenae and P. coronata avenae occurred best at 10–30^oC and at 20^oC respectively. The optimum temperature for germ-tube growth and for appressorial formation was 20^oC for both rusts. An inverse relationship was observed between light intensity and prepenetration development with maximal germination of uredospores, germ-tube growth and appressorial formation occurring in darkness. Under optimum conditions maximum percentage germination and appressorium formation of both rusts was attained within 4 and 12 h after inoculation respectively. The proportion of germinated uredospores of crown rust which gave rise to appressoria was about twice that observed for stem rust. No significant differences were observed in prepenetration development between the single and mixed race inocula of the two rusts. Although germination of uredospores was significantly greater on water agar than on oat leaves, there were no significant differences in prepenetration development of the rusts on the various oat cultivars and species examined. Consequently, the data failed to indicate the presence of resistance mechanisms operating during the prepenetration phase of the infection process on the cultivars and species examined.     

Effect of light intensity on sporulation of Botryosphaeria ribis

Pycnidia were produced by six of seven isolates ofB. ribis at one or more intensity levels of continuous illumination at 21 °C. Under conditions of alternating light (12 h–27 °C) and darkness (12 h–21 °C) pycnidia formed in cultures of six isolates at three or more intensity levels, while one isolate failed to form pycnidia at any intensity level. Pycnidia did not develop when cultures were incubated in complete darkness. Exposure periods as brief as 2 days under continuous illumination at 21 °C induced pycnidial formation. In alternating light (12 h–27 °C) and darkness (12 h–21 °C), the shortest period of exposure which induced pycnidial formation was 4 days. Continuous illumination at 21 °C favored development of uniloculate pycnidia, while alternating light (12 h–27 °C) and darkness (12 h–21 °C) favored formation of multiloculate pycnidia.Contribution No. 22 from The Botany Section of The Department of Biology, The Pennsylvania State University.     

spoVIC, a new sporulation locus in Bacillus subtilis affecting spore coats, germination and the rate of sporulation

A mutation near cysB on the Bacillus subtilis chromosome marks a new sporulation locus, spoVIC. It causes spores to germinate more slowly than those of the wild-type under all conditions and, from indirect evidence, it does not appear to alter the affinity for the germinant L-alanine. The mutant spores have some deficiency of coat proteins (particularly the alkalisoluble coat protein, Mr = 12 000) and the spore coat layers are disorganized. The mutant strain grows normally and sporulates normally until stage II, after which its sporulation is delayed by about 2 h compared to that of the wild-type. This delay results in the prolonged synthesis of some coat proteins and the late synthesis of others. The abnormal coat may be the cause of the germination deficiency. A double mutant strain carrying the spoVIC610 mutation together with gerE36 sporulates slowly. Its spores have very little coat protein, are sensitive to heat, lysozyme and organic solvents, but germinate as well as the strain carrying the spoVIC mutation alone. The role of the spore coat in germination is discussed in the light of these findings.     

艰难类梭菌芽胞形成和萌发相关调控的研究进展

艰难类梭菌(Clostridioides difficile)是一种革兰氏阳性、可产毒素的专性厌氧菌,是引起抗生素相关性腹泻的主要致病菌.芽胞是造成艰难类梭菌传播和感染复发的重要因素,其形成和萌发在感染的发展过程中起到重要作用.近年来,越来越多的艰难类梭菌芽胞形成和萌发的具体机制被阐明.本文就近年来艰难类梭菌芽胞形成和...     

Inducing sporulation in Alternaria solani II. Effect of light

Among the three different light sources viz. incandescent electric light, infra-red light and sunlight, only sunlight was effective in inducing sporulation in petri dish cultures ofA. solani. The intensity of sporulation depended upon the age and growth stage of the cultures, duration and number of exposures to light and the presence or absence of a sporulating zone in the culture. Maximum sporulation was obtained in the case of 6 days old partially grown cultures by inducing the formation of sporulating zone which appeared in 24 hours after every exposure of 60 minutes to sunlight.     

Effect of light and temperature on the germination of lettuce seeds

A short period (15–30 min) at 30° C promotes germination of seeds of Lactuca sativa L. cv. Repolhuda in darkness. Far-red light reverses this stimulation, and the escape curves for phytochrome and high-temperature action are quite similar, indicating that the two factors act at a common point in the chain of events leading to germination. It is suggested that high temperature acts by decreasing the threshold of the active, far-red absorbing, form of phytochrome (Pfr) needed to promote germination.Abbreviations FR far-red light - Pfr far-red-absorbing form of phytochrome - R red light     

Effect of light on seed germination and seedling growth ofMerremia species

The seeds ofMerremia aegyptia are indifferent to light conditions for germination, which becomes 100% in the first 24 hrs. On the contrary, inM. dissecta the highest percentage was found in red light within the first 24 hrs. This difference was lost later on except in far-red. The seedling growth of the two species was least in blue and red lights, respectively. However, the growth of hypocotyl in the former species was faster than the latter.     

光照、温度和pH值对小黑桫椤孢子萌发及早期配子体发育的影响

小黑桫椤(Alsophila metteniana)被列为我国国家二级保护植物。为探讨其种群数量下降原因, 作者采用无菌培养方法和显微观察技术, 在实验条件下研究了光照强度、光质、温度和pH值对小黑桫椤孢子萌发及早期配子体发育的影响。结果表明: 孢子萌发和配子体发育的最适光照强度为40–70 µmol;m^–2;s^–1, 全黑暗时孢子不萌发;白光、红光、黄光和蓝光下的萌发率分别为68.78%、65.66%、63.74%和7.51%; 白光和蓝光下配子体可以形成正常的心形原叶体, 红光和黄光下配子体发育一直处于丝状体阶段。孢子萌发和配子体发育需要在酸性土壤进行(pH值在3.7–6.7); 孢子萌发的适宜温度为20–30℃。从孢子接种到心形原叶体形成需要55 d左右。根据上述结果, 我们认为光照强度是小黑桫椤孢子萌发的必需条件, 光质是限制小黑桫椤孢子繁殖的重要原因; 光质、温度和pH值等环境因子的作用和配子体发育时间等是导致小黑桫椤种群数量日趋减少的主要原因。     

Effect of in situ storage,light, and moisture on the germination of two wetland tropical trees

Seed germination was evaluated for Annona glabra L. and Pachira aquatica Aubl. in the wetland conditions of La Mancha, Veracruz, Mexico. These species have recalcitrant seeds and hydrochoral dispersal. Germination experiments were carried out under varying moisture (high, middle, and low) and light (below canopy and open sky) levels as well as after being stored in contrasting natural conditions. Seeds were stored both floating in water and buried in wetland soil for 15, 30, 60, and 90 days. P. aquatica seeds germinated faster in low and medium moisture, regardless of light intensity. After 45 days, for example, they exhibited 87–73% germination in medium-moisture/canopy and low-moisture/canopy treatments, respectively. In high moisture, seeds reached similar percentages after 60 days (80%). Storage by burial caused the death of seeds regardless the time they spent underground, while those stored in water germinated at a rate of over 90%. A. glabra seeds germinated better (98%) in low-moisture/sunny conditions. As to storage, they responded favorably to both burial and water techniques but germinated more readily in treatments that involved a long storage period. Evaluation of the germination behavior of A. glabra and P. aquatica seeds subject to varying in situ storage and germination conditions illustrates the response capacity of each species during early phases of development.     

Oxygen delivery requirements of Colletotrichum truncatum during germination,vegetative growth,and sporulation

Optimization of O₂ delivery was the key to successful conidiation of Colletotrichum truncatum in submerged fermentor cultures supplied with 20 g carbon/l and C:N at the optimal 10:1 mass ratio for spore efficacy. Minimal mycelial fragmentation and maximal biomass and spore yields were provided by an O₂ transfer program that called for gradual increases in stirring rate to compensate for rising cell concentration and viscosity. The utility of an event-based O₂ transfer program was further supported by our observation of different O₂ requirements for each phase of the life cycle. Spore germination did not occur in cultures sparged with N₂. However, even low levels of O₂ [10% dissolved O₂ tension (DOT)] allowed 100% germination. The specific growth rate of the mycelia was a Monod-like function of DOT. The maximal growth rate was achieved when 15% DOT was provided via O₂ transfer at a specific rate of 5.4 × 10^–3 mol/g per hour. Sporulation had a strict O₂ requirement, and its rate and yield were optimized by providing 55% DOT following the cessation of growth. The specific O₂ demand of optimally sporulating mycelia was 4.9 × 10^–4 mol/g per hour, an order of magnitude less than that associated with growing mycelia. Behaving as a pseudoplastic fluid, the fermentation broth reached a maximum apparent viscosity of 70 P at the onset of sporulation when the O₂ demand was low. However, the maximum power requirement approx. 7.9 W/l occurred during the last 36 h of growth when the O₂ demand was highest. Correspondence to: P. J. Slininger