A comparison of Fusarium avenaceum and Fusarium caeruleum as causes of wastage in stored potato tubers

Fusarium avenaceum is reported for the first time as a cause of rotting of potato tubers in Britain. The progress of rotting in tubers infected with F. avenaceum has been compared with dry rot due to F. caeruleum in the laboratory, clamp and potato store. Of the four varieties, Majestic, King Edward, Doon Star and Arran Pilot, tested for susceptibility, King Edward was the most susceptible to F. avenaceum and Doon Star to F. caeruleum.
Optimum temperatures for growth on potato-dextrose agar were 20-25 C. for F. avenaceum and 20 C. for F. caeruleum ; maximum temperatures were > 30 and 30 C. respectively. For infection of wounded potato tubers, cardinal temperatures for F. avenaceum were similar to those on agar, but for F. caeruleum the optimum was 15 C. and the maximum 25 C. The optimum temperature for rotting tended, with both species, to be higher in the more susceptible potato varieties. At low temperatures F. caeruleum caused quicker rotting than did F. avenaceum , even though its rate of growth on agar was scarcely more than half that of the latter.
High humidity favoured rotting especially by F. avenaceum; F. caeruleum was more tolerant of relatively low humidity. Both species caused quicker rotting in the clamp than in store, even though there was no appreciable difference in mean temperature between the clamp and the store. This was attributed to the higher atmospheric humidity in the clamp.     

Effects of delaying fungicide treatment of wounded potatoes on the incidence of Fusarium dry rot in store

Potato tubers artificially inoculated with Fusarium solani var. coeruleum or F. sulphureum 3 months after harvest were uniformly wounded and held at 5, 10 or 15°C for up to 21 days before immersion in fungicide suspensions. Holding tubers for 14 days at 15°C (curing conditions) or at 5°C did not alter the incidence of dry rot subsequently developing on tubers stored at 10°C, and holding tubers for up to 21 days at 15°C slightly increased disease caused by both pathogens. Thiabendazole, imazalil and prochloraz applied to tubers immediately after wounding almost completely prevented dry rot. Treatment after 3 days was less effective and the amount of disease increased with further delay; fungicides were more effective on tubers held at 5°C than at 10 or 15°C before treatment and storage, and efficacy of the fungicide was decreased by increasing the amount of inoculum on tubers. Wounds became less susceptible to infection by F. solani var. coeruleum and F. sulphureum when tubers were held at 15°C before inoculation, and the incidence of rots was decreased by 70–80% by delaying inoculation for 7 days. Treating tubers with dichlorophen immediately after wounding slightly increased the disease. The effects of fungicide treatment, curing conditions and wound healing are discussed.     

指状青霉Penicillium digitatum侵染柑橘果实的细胞学研究

采用电子显微镜技术系统研究了指状青霉Penicillium digitatum对柑橘果实的侵染过程及超微结构特征。结果表明室温条件下,接种12h后,伤口附近的分生孢子开始萌发产生芽管;然后从伤口处直接侵入果实表皮细胞内;接种24h后,受侵染果实细胞中的菌丝向相邻细胞扩展蔓延,寄主细胞壁开始消解,质壁分离,细胞内含物及各类细胞器凝集,颜色加深,最后完全消解,伤口部位的果皮开始变软,伤口处的菌丝向外生长;84h后伤口处病斑软化,部分长出白色的霉层;96h后病斑软化面积直径达3cm,白色霉层面积逐渐扩大;120h后白色霉层中间伤口处霉层颜色加深变为灰绿色;144h后整个果实变软腐烂。果胶质标记结果表明,菌丝侵入果实后产生果胶酶并降解柑橘细胞壁中的果胶,使得细胞壁松弛,软化腐烂。     

立枯丝核菌(Rhizoctonia solani)对日本结缕草(Zoysia japonica steud.)的侵染过程研究

【目的】了解立枯丝核菌(Rhizoctonia solani) AG1 IA对日本结缕草(Zoysia japonica steud.)的侵染过程及其引起的病害症状,为进一步从分子水平研究该菌的病理学侵染机制和草坪草抗病分子育种提供理论基础。【方法】通过根部接种法促使立枯丝核菌与日本结缕草无菌苗建立侵染关系,从而对其病叶率、病株率及病情指数进行统计分析,同时结合组织染色透明技术及植物组织石蜡切片对立枯丝核菌的侵染过程、感染方式进行研究。【结果】立枯丝核菌AG1 IA的侵染过程主要为：菌丝吸附在植物组织表面,并沿组织表面定向生长,形成侵染结构——侵染垫与组织建立密切的侵染关系。菌丝通过细胞间隙侵入植物组织内部,主要侵染植物皮层细胞及除木质部导管以外的整个维管束系统。结缕草的地上部分与地下部分组织对立枯丝核菌的侵染显现不同的寄主反应。【结论】立枯丝核菌的侵染过程主要包括吸附、定向生长、渗透、定殖4个部分;立枯丝核菌的侵染主要引起结缕草叶片病症;结缕草病变与菌丝直接侵染无直接联系,表明该菌具有复杂的侵染机制。     

Hydrolytic Activities of Fusarium solani and Fusarium solani f. sp. eumartii Associated with the Infection Process of Potato Tubers

The development of dry rot caused by Fusarium solani f. sp. eumartii was evaluated in susceptible (Huinkul) and resistant (Spunta) potato cultivars. Fungal proteolytic and polygalacturanase activities were measured at different days postinoculation either with the pathogenic F. solani f. sp. eumartii, isolate 3122 or with the non‐pathogenic F. solani, isolate 1042. After inoculation with the pathogenic fungus, proteolytic and polygalaturonase activities were higher in the susceptible than in the resistant cultivar. In addition, we found a correlation between the levels of proteolytic activity detected in the intercellular washing fluids with the size of the lesion area caused by F. solani f. sp. eumartii in Huinkul tubers. The action of the proteolytic activity over cell wall proteins of both potato cultivars was assayed. An extracellular potato protein with homology to proteinase inhibitors of the Kunitz family was identified as a substrate of the proteolytic activity in the susceptible cultivar. A microscopic study revealed differences between the potato genotypes in the rate of response to infection by F. solani f. sp. eumartii. In addition, the cell wall alteration caused by F. solani f. sp. eumartii in cortical cells of susceptible tubers was evaluated. The data with respect to the correlation between the course of cyto‐ and biochemical events of the two host–pathogen interactions were discussed.     

On the Occurrence of Intracellular Blue-green Algae in Cortical Cells of the Apogeotropic Roots of Macrozamia communis L. Johnson

The occurrence of species of the cyanophytes Nostoc and Anabaenain the cortex near the algal zone is reported for apogeotropicroots of Macrozamia communis L. Johnson. Algae were found tooccur both intercellularly and intracellularly in cells of theinner and outer cortex. This is the first record of intracellularalgae in the cycads and only the second report of this phenomenonin vascular plants. By examination of cells at various stagesof invasion by algae, it is interpreted that algal invasionof cortical cells and intercellular spaces is preceded by mucusapparently secreted by algal zone cells of the host, and depositedin intercellular spaces of cortical parenchyma cells nearby.Also algal penetration of cortical cells is preceded by an algalinvasion front of finely granular mucal material which bypassesmucus already deposited in intercellular spaces and may eitherlyse part of the host cell wall or enter through the plasmo-desmata,filling much of the cell cavity. Subsequently, large numbersof the algal symbionts enter the cell and may be enclosed withinhost wall material. Electron microscopic techniques are nowbeing employed to further clarify these invasion processes.     

Wound-induced superoxide production and PAL activity decline with potato tuber age and wound healing ability

Wound healing of potato tubers involves the concerted action of several enzymes that facilitate polymerization of phenolics into suberin at the wound site. A decline in the efficiency of healing and resistance to pathogens with advancing tuber age was associated with reduced ability of older tubers to produce superoxide radicals (FRs) in response to wounding. Autophotographs of luminol‐treated longitudinal sections of tissue from 6‐, 18‐ and 30‐month‐old tubers revealed a substantial decline in superoxide production at the wound surface with advancing age. Older tubers were less able to respond to wounding by increasing phenylalanine ammonia lyase (PAL) activity. This enzyme produces t‐cinnamic acid, which constitutes a component of the phenolic domain of suberin, and is normally induced by wounding and/or ethylene. Interestingly, the ability of wounded tissue to oxidize exogenous 1‐aminocyclopropane‐1‐carboxylic acid (ACC) to C₂H₄ also decreased with advancing tuber age. The oxidation of ACC was inhibited by the FR scavenger, n‐propyl gallate (PG), and inhibition was greatest in tissue from younger tubers, reflecting their greater ability to produce superoxide radicals upon wounding. Regardless of tuber age, 1‐aminocyclobutane‐1‐carboxylic acid, an ACC oxidase inhibitor, did not inhibit C₂H₄ generation from exogenous ACC. Hence, C₂H₄ production from ACC by wounded tuber tissue is largely non‐enzymatic and FR‐driven, and thus serves as an indicator of the ability of wounded tissue to produce superoxide. Age‐induced reduction in PAL activity and FR production at the wound surface probably limited the oxidative polymerization of phenolics into suberin during wound periderm formation. The age‐induced loss in ability of wounded tissue to heal and resist pathogens is thus consistent with reduced synthesis and polymerization of phenolic adducts into suberin, a consequence of reduced FR and PAL activity at the wound surface.     

红豆草根瘤侵入线的超微结构研究

用透射电镜对红豆草根瘤侵入线的超微结构进行了观察研究.结果表明,(1)红豆草根瘤侵入线由胞间隙和胞间层细胞壁内陷形成,它们的体积较小,多为管状,基质丰富,含菌很少,常有分叉和1个以上的基质区,而且不同基质区的电子密度、细菌数量和侵入线壁厚度都不相同.(2)红豆草根瘤的侵入线十分丰富,它们不仅大量存在于根瘤分生细胞和幼龄侵染细胞中,也经常出现在发育成熟的侵染细胞内.(3)红豆草根瘤中有一种近似圆形的特殊结构,表面由一层膜包围,其内电子密度较低且无固定结构,且只位于侵染细胞的细胞质中,常在侵入线附近,从不出现在侵染细胞的液泡内和非侵染细胞里面.     

Early development ofRhizobium-induced root nodules ofParasponia rigida. I. Infection and early nodule initiation

Summary The first of two major steps in the infection process in roots ofParasponia rigida (Ulmaceae) following inoculation byRhizobium strain RP501 involves the invasion ofRhizobium into the intercellular space system of the root cortex. The earliest sign of root nodule initiation is the presence of clumps of multicellular root hairs (MCRH), a response apparently unique amongRhizobium-root associations. At the same time or shortly after MCRH are first visible, cell divisions are initiated in the outer root cortex of the host plant, always subjacent to the MCRH. No infection threads were observed in root hairs or cortical cells in early stages. Rhizobial entry through the epidermis and into the root cortex was shown to occur via intercellular invasion at the bases of MCRH. The second major step in the infection process is the actual infectionper se of host cells by the rhizobia and formation of typical intracellular infection threads with host cell accommodation. This infection step is probably the beginning of the truly symbiotic relationship in these nodules. Rhizobial invasion and infection are accompanied by host cortical cell divisions which result in a callus-like mass of cortical cells. In addition to infection thread formation in some of these host cortical cells, another type of rhizobial proliferation was observed in which large accumulations of rhizobia in intercellular spaces are associated with host cell wall distortion, deposition of electron-dense material in the walls, and occasional deleterious effects on host cell cytoplasm.     

Systemic Fungal Growth of Fusarium culmorum in Stems of Winter Wheat

Abstract Systemic fungal growth of Fusarium culmorum in winter wheat was investigated under conditions precluding secondary infections by water splash. Growth of F. culmorum in stem tissue was found in both wounded and soil inoculated plants with both methods resulting in a high level of infection. Crown rot can therefore lead to infection of the higher stem internodes under conditions not suitable for Fusarium dispersal. However, no evidence was found for systemic fungal growth leadingto infected heads. Existence of genetic variation for resistance to spread of F. culmorum in the host was found. This resistance was not correlated with resistance to Fusarium head blight.     

Vegetative anatomy of subtribe Orchidinae (Orchidaceae)

Leaves in Orchidinae are essentially glabrous; anticlinal walls of foliar epidermal cells arc basically straight-sided to curvilinear, and cells arc fundamentally polygonal on both surfaces; adaxial cells are larger than abaxial cells. Stomata arc anomocytic and usually only abaxial and superficial; substomatal chambers are small to moderate; outer and inner guard cell ledges are mostly small. There is no hypodermis nor are there fibre bundles. Mesophyll is homogeneous, chlorcnchyma cells arc thin-walled, and intercellular spaces numerous. Crystalliferous idioblasts abound. Vascular bundles are collateral, organized in a single series. and lack associated sclerenchyma. Bundle sheath cells are thin-walled and chlorophyllous. Stems are glabrous; stomata arc frequent in one species, lacking in others. Cortical cells are oval to circular, thick-walled, and interspersed with triangular intercellular spaces. Ground-tissue cells are circular, and triangular intercellular spaces are present. Vascular bundles arc collateral and scattered throughout the ground-tissue or are absent from the central ground-tissue. Epidermis in absorbing roots is one-layered and non-velamcntous. Exodcrmal cells are thin-walled and dead cell walls bear tenuous scalariform bars; some species lack an exodermis. Outer cortical cells are polygonal and lack intercellular spaces; middle layer cortical cells are rounded with triangular intercellular spaces; inner layer cells are polygonal and lack intercellular spaces. Endodermis and pericycle are thin-walled and one-layered. Vascular cylinder is mostly 7–9-arch with xylcm and phloem components alternating regularly; vascular tissue is embedded in parenchyma; pith cells are parenchymatous, polygonal, thin-walled and lack intercellular spaces. Root tubers generally bear a velamen of variable thickness; bulbous-based unicellular hairs frequently form a dense mat; exodermal cells are thin-walled; dead cells have scalariform bars, passage cells are sparse. Ground-tissue consists of rounded water-storage and assimilatory cells interspersed with triangular or quadrangular intercellular spaces; peripheral cells arc polygonal lacking intercellular spaces. Vascular tissue consists of monarch to pentarch meristeles distributed thoughout the ground-tissue each surrounded by a uniscriale endodermis of thin-walled cells. Thin roots ofPlalanthera exhibit a typical central cylinder surrounded by a homogeneous cortex uninterrupted by meristeles; thicker roots show a central vascular cylinder and cortex in which meristeles are also present; in globoid root tubers there is no central cylinder, and the ground-tissue is replete with scattered meristeles. Because the central vascular cylinder in Platanthera gives rise to branches (meristeles), these represent components of a single vascular system and are not separate stelar entities as implied by the use of the term ‘polystele’.     

The Nature of Systemic Invasion of Stem and Leaves of Sunflowers by Plasmopara helianthi Novot. var. helianthi Novot. with Mechanism of Sporulation and Zoospore Release

During the systemic development of Plasmopara helianthi Novot. var. helianthi Novot. the hyphae in the stem advance especially through the intercellular spaces of loose parenchyma forming haustoria in adjacent cells. The hyphae which reach leaf blades through petioles continue their growth along the main veins intercellularly in non-vascular tissues enclosing the vascular bundles. At the same time, the hyphae spread to intercellular spaces of spongy parenchyma where the growth is limited by the veinlets resulting in angular chlorotic lesions. Under humid conditions the sporangiophores arising from the aggregated hyphae in a substomatal cavity emerge through the stomatal pore on the lower side of the leaf and zoosporangia are borne terminally on sporangiophores. Soon after biflagellated zoospores are liberated into distilled water from the zoosporangia, they retract their flagellae and then lyse.     

The vascular plant‐pathogenic bacterium Ralstonia solanacearum produces biofilms required for its virulence on the surfaces of tomato cells adjacent to intercellular spaces

The mechanism of colonization of intercellular spaces by the soil‐borne and vascular plant‐pathogenic bacterium Ralstonia solanacearum strain OE1‐1 after invasion into host plants remains unclear. To analyse the behaviour of OE1‐1 cells in intercellular spaces, tomato leaves with the lower epidermis layers excised after infiltration with OE1‐1 were observed under a scanning electron microscope. OE1‐1 cells formed microcolonies on the surfaces of tomato cells adjacent to intercellular spaces, and then aggregated surrounded by an extracellular matrix, forming mature biofilm structures. Furthermore, OE1‐1 cells produced mushroom‐type biofilms when incubated in fluids of apoplasts including intercellular spaces, but not xylem fluids from tomato plants. This is the first report of biofilm formation by R. solanacearum on host plant cells after invasion into intercellular spaces and mushroom‐type biofilms produced by R. solanacearum in vitro. Sugar application led to enhanced biofilm formation by OE1‐1. Mutation of lecM encoding a lectin, RS‐IIL, which reportedly exhibits affinity for these sugars, led to a significant decrease in biofilm formation. Colonization in intercellular spaces was significantly decreased in the lecM mutant, leading to a loss of virulence on tomato plants. Complementation of the lecM mutant with native lecM resulted in the recovery of mushroom‐type biofilms and virulence on tomato plants. Together, our findings indicate that OE1‐1 produces mature biofilms on the surfaces of tomato cells after invasion into intercellular spaces. RS‐IIL may contribute to biofilm formation by OE1‐1, which is required for OE1‐1 virulence.     

Initial stages in the morphogenesis of nitrogen-fixing stem nodules of Sesbania rostrata.

Morphogenesis of stem nodules in Sesbania rostrata was studied over a period of 6 days after inoculation with an appropriate species of Rhizobium. Nodulation sites were initially slightly raised, circular areas 0.3 to 0.6 mm in diameter and 4 to 5 mm apart in vertical rows along the length of the stem. Each site was underlaid by an adventitious root primordium. A site became susceptible to infection by a specific Rhizobium sp. when the root primordium broke through the epidermis, leaving a fissure. Rhizobia multiplied within this fissure and colonized the exposed intercellular spaces. The infection extended inward as narrow, branched intercellular threads moved into a cortical meristematic zone, where cell division was initiated, and invagination of infection thread branches into adjacent plant cells followed. Rhizobia were released into the plant cells and surrounded immediately by plant membrane. Intracellular rhizobia divided actively, leading to bacteroid-filled cells. Infected areas enlarged and coalesced as the nodule matured.     

DRY-ROT DISEASE OF THE POTATO

A laboratory method is described in which a modified hypodermic syringe is used to inoculate potato tubers with Fusarium caeruleum. This injection method gives consistent results and permits reliable assessments of factors such as varietal susceptibility.
The occurrence of arrested lesions is noted.
Suspensions of very high spore concentration or of spores already germinated did not greatly increase the amount of dry rot above that given by the standard too spores per inoculation, provided that spore germination was adequate.
The size of tuber and site of inoculation were found to have a considerable effect upon the results of inoculation experiments, large tubers being more susceptible than small ones and the heel end being more susceptible than the rose end. The necessity for uniformity of material used in inoculation experiments is emphasized.     

Evaluation of Spring and Winter Wheat Reaction to Fusarium culmorum and Fusarium avenaceum

Winter (37), spring (8) wheat accessions and additionally, 7 double haploid (DH) lines were examined for susceptibility to Fusarium seedling blight after inoculation with F. culmorum and F. avenaceum. Winter accessions exhibited lower susceptibility of about 30% to both pathogens than spring cultivars. Susceptibility of winter cultivars varied from low (22%) to high (97%). Evaluation of the root was found to be more reliable than evaluation of coleoptile necrosis.
F. avenaceum infected mostly root and, to a lesser extent, coleoptile and leaves, with about a three times lower disease score of coleoptile against root. F. culmorum caused a 1.5 higher disease score on root than on coleoptile. Susceptibility of DH lines was different from susceptibility of parental forms. Reaction of individual accessions to F. culmorum and F. avenaceum was different.     

Fluid transport and the dimensions of cells and interspaces of living Necturus gallbladder

The volume of the cells and lateral intercellular spaces were measured in living Necturus gallbladder epithelium. Under control conditions, the volume of the lateral spaces was 9% of the cell volume. Replacement of mucosal NaCl by sucrose or tetramethylammonium chloride (TMACl) caused intercellular spaces to collapse. During mucosal NaCl replacement, cell volume decreased to 79% of its control value. When NaCl was reintroduced into the mucosal bath, the intercellular spaces reopened and the cells returned to control volume. The NaCl active transport rate, calculated from the rate of cell volume decrease, was 266 pM/cm2.s, close to the observed rate of transepithelial salt transport. It was calculated from the decrease in cell volume that all of the intracellular NaCl was transported out of the cell during removal of mucosal NaCl. The flux of salt across the apical membrane, calculated from the rate of cell volume increase upon reintroducing mucosal NaCl, was 209 pM/cm2.s, in good agreement with estimates by other methods. The electrical resistance of the tight junctions was estimated to be 83.9% of the total tissue resistance in control conditions, suggesting that the lateral intercellular spaces normally offer only a small resistance to electrolyte movement.