Localized Melanization of Appressoria Is Required for Pathogenicity of Venturia inaequalis

ABSTRACT During formation of appressoria produced from conidia and ascospores of Venturia inaequalis, a dark brown ring structure was detected at the base of appressoria. This melanized appressorial ring structure (MARS) was attached to the leaf surface like a sealing ring and formed the fungus-plant interface; it is believed to be required for pathogen penetration of the cuticle. Neither germ tubes nor infection structures beneath the cuticle were found to be visibly melanized. MARS were formed not only on apple leaves but also on leaves of nonhost plants and artificial surfaces differing in hydrophobicity; the formation of appressoria and MARS was confined to hard surfaces. The melanin nature of the ring was confirmed by using melanin biosynthesis inhibitors. Applications prior to inoculation largely inhibited the melanization and reduced infection rate by 45 to 80%; curative applications were not effective. Transmission electron microscopy verified a localized melanization of the cell wall around the penetration pore, and melanin was incorporated into all layers of the fungal cell wall. Appressoria without MARS were not able to infect the plant, suggesting that this structure can be considered to be a pathogenicity factor in V. inaequalis.     

Conidial germination, infection structure formation, and early colony development of powdery mildew on poinsettia

ABSTRACT Powdery mildew disease on poinsettias (Euphorbia pulcherrima) growing in commercial greenhouses was first observed in the United States in 1990 and has become an economically significant problem for poinsettia growers in the Midwest and northern United States since 1992. The temporal development of infection structures produced by conidial germ tubes of the pathogen (Oidium sp.) and the effect of high temperature on their development were investigated using poinsettia leaf disks placed in humidity chambers. Observations were made using light microscopy and scanning electron microscopy. At 20 degrees C (85% relative humidity), conidia germinated and formed an appressorium within 6 h of inoculation. Germination over time followed a monomolecular curve (r(2) = 0.77, P 相似文献   

疏水表面诱导橡胶树炭疽菌侵染结构的发育分化过程

为了解橡胶树2种炭疽病菌的侵染结构发育分化过程,采用平板菌落生长速率法测定了3株胶孢炭疽菌Colletotrichum gloeosporioides和3株尖孢炭疽菌C.acutatum的菌丝生长速率,测量其分生孢子大小,显微观察2种炭疽菌在疏水表面诱导下侵染结构的发育分化过程。结果表明,胶孢炭疽菌菌丝生长速率为0.96~1.36 cm/d,显著高于尖孢炭疽菌的菌丝生长速率0.72~0.89 cm/d,但二者分生孢子大小无显著差异。在疏水表面诱导下,2种炭疽菌分生孢子在接种2~6 h后开始萌发,12 h孢子萌发率为71.70%~88.05%,13~16 h开始分化附着胞,24 h附着胞形成率为48.99%~70.74%,36 h菌丝诱发形成大量附着枝,48 h后分生孢子产生的次生菌丝也可诱发形成附着枝,附着枝呈圆形、姜瓣形、梨形或不规则形。分生孢子极易产生,可在菌丝顶端成簇或菌丝侧面排列产生,也可由分生孢子形成的芽管产生,或在芽管分化附着胞过程分枝形成分生孢子;附着胞多着生于芽管顶端,少数附着胞顶端可继续萌发类似短芽管结构,再次分化形成可黑色化的次级附着胞。表明橡胶树2种炭疽菌不同菌株间分生孢子萌发时间、孢子萌发率、附着胞形成时间和形成率有一定差异,但种间无明显差异;橡胶树炭疽菌分生孢子极易形成,在疏水表面容易分化形成附着胞和附着枝,说明具有极强的适生性。     

Effect of temperature,relative humidity,leaf wetness and leaf age on <Emphasis Type="Italic">Spilocaea oleagina</Emphasis> conidium germination on olive leaves

The effects of temperature, relative humidity (RH), leaf wetness and leaf age on conidium germination were investigated for Spilocaea oleagina, the causal organism of olive leaf spot. Detached leaves of five ages (2, 4, 6, 8 and 10 weeks after emergence), six different temperatures (5, 10, 15, 20, 25 and 30°C), eight wetness periods (0, 6, 9, 12, 18, 24, 36 and 48 h), and three RH levels (60, 80 and 100%) were tested. Results showed that percentage germination decreased linearly in proportion to leaf age (P < 0.001), being 58% at 2 weeks and 35% at 10 weeks. A polynomial equation with linear term of leaf age was developed to describe the effect of leaf age on conidium germination. Temperature significantly (P < 0.001) affected frequencies of conidium germination on wet leaves held at 100% RH, with the effective range being 5 to 25°C. The percent germination was 16.1, 23.9, 38.8, 47.8 and 35.5% germination at 5, 10, 15, 20 and 25°C, respectively, after 24 h. Polynomial models adequately described the frequencies of conidium germination at these conditions over the wetness periods. The rate of germ tube elongation followed a similar trend, except that the optimum was 15°C, with final mean lengths of 175, 228, 248, 215 and 135 μm at 5, 10, 15, 20 and 25°C, respectively after 168 h. Polynomial models satisfactorily described the relationships between temperature and germ tube elongation. Formation of appressoria, when found, occurred 6 h after the first signs of germination. The percentage of germlings with appressoria increased with increasing temperature to a maximum of 43% at 15°C, with no appressoria formed at 25°C after 48 h of incubation. Increasing wetness duration caused increasing numbers of conidia to germinate at all temperatures tested (5–25°C). The minimum leaf wetness periods required for germination at 5, 10, 15, 20 and 25°C were 24, 12, 9, 9 and 12 h, respectively. At 20°C, a shorter wetness period (6 h) was sufficient if germinating conidia were then placed in 100% RH, but not at 80 or 60%. However, no conidia germinated without free water even after 48 h of incubation at 20°C and 100% RH. The models developed in this study should be validated under field conditions. They could be developed into a forecasting component of an integrated system for the control of olive leaf spot.     

Rapid pregermination and germination responses of <Emphasis Type="Italic">Erysiphe pisi</Emphasis> conidia to contact and light

In darkness, most Erysiphe pisi conidia responded rapidly to contact with a hydrophobic artificial substratum and released extracellular material (ECM) in the same way as on pea cuticle. On this substratum and barley leaf epidermis, conidia then produced a germ tube that emerged close to the substratum, contacted it, and differentiated an appressorium. By contrast, on a hydrophilic substratum, ECM release and germination were delayed and infrequent, and germ tubes often emerged and faced away from the substratum toward vertical light, thereby failing to make contact and form appressoria. This finding supported the hypothesis that ECM release is involved in both triggering germination and sensing substratum contact. Exposure to white light dramatically affected the germ tube emergence site so most emerged from a site in the conidial wall facing the light. Lateral light did not affect the frequency of germ tubes making substratum contact; but when lit from above, most germ tubes emerged up, facing away from the substratum. The germ tubes formed in light were longer than those formed in darkness, but no phototropism was found for the elongating tubes. Examination of Blumeria graminis indicated that its conidia and germ tubes are insensitive to white light.     

苹果炭疽叶枯病病原Glomerella cingulata及其侵染过程

为明确苹果炭疽叶枯病病原菌围小丛壳Glomerella cingulata的侵染致病特征,在分离获得该病原菌的基础上,采用形态学观察、ITS序列分析和致病性测定对其进行了鉴定,并利用光学和扫描电子显微镜对病原菌在嘎啦苹果叶片上的侵染过程进行了研究.结果表明,在陕西咸阳地区分离获得的9株病原菌均为围小丛壳G.cingulata.25 ℃下接种9 h后,分生孢子中间产生隔膜,双胞化,并萌发产生芽管和附着胞;24 h后分生孢子的2个细胞均可萌发并形成芽管及附着胞,部分芽管顶端可产生次级分生孢子;48 h后次级分生孢子萌发形成附着胞;72 h后,附着胞下形成的侵染钉可直接入侵寄主,在表皮细胞内形成初生菌丝和次生菌丝,此时叶片表面已出现褐色斑点.接种7 d后叶片病斑处出现分生孢子盘和子囊壳.表明陕西省近年出现的苹果炭疽叶枯病病原菌为围小丛壳G.cingulata,该病菌在嘎啦叶片上的一些特殊侵染行为可能是导致该病害易在短时间内暴发的重要原因.     

影响柿树炭疽菌孢子萌发、附着胞形成及致病性的环境因子

 在凹玻片上测定了不同浓度的葡萄糖溶液对柿树炭疽菌(Colletotrichum gloeosporioides)的分生孢子萌发和附着胞形成率的影响,结果表明:分生孢子萌发率随葡萄糖浓度升高而增加,但附着胞形成率下降,芽管长度增加,附着胞的直径几乎没有变化;随着时间的延长,孢子萌发率和附着胞形成率都有所增加。不同pH对分生孢子萌发和附着胞形成率的影响结果显示,分生孢子在pH 2.0~9.0的溶液中可以萌发,并产生附着胞;最适分生孢子萌发和附着胞形成的pH是在5.0~6.0。不同pH处理的致病试验结果说明。在23℃时病斑在pH 4.0~8.0条件下都可产生;致病试验结果也发现,17℃时,pH6.0处理能发病,但不形成分生孢子团,pH 5.0处理不发病;15℃时,pH 5.0和pH 6.0的处理都不能发病。温度对菌丝生长的影响显示,菌落生长最适温度是25℃左右,高温抑制菌落生长。寄主表面侵染结构扫描电镜观察结果表明,芽管长度变化很大,芽管可以纵向沿着脊或沟延伸,也可横向通过脊沟;附着胞均在沟底或近底部形成。     

Pathogenicity of <Emphasis Type="Italic">Mycochaetophora gentianae</Emphasis>, causal fungus of gentian brown leaf spot,as affected by host species,inoculum density,temperature, leaf wetness duration,and leaf position

When the influence of host species, inoculum density, temperature, leaf wetness duration, and leaf position on the incidence of gentian brown leaf spot caused by Mycochaetophora gentianae, was examined, the fungus severely infected all seven Gentiana triflora cultivars, but failed to infect two cultivars of G. scabra and an interspecific hybrid cultivar. Inoculum density correlated closely with disease incidence, and a minimum of 10² conidia/mL was enough to cause infection. In an analysis of variance, temperature and leaf wetness duration had a significant effect upon disease incidence, which increased with higher temperature (15–25°C) and longer duration of leaf wetness (36–72 h). No disease developed at temperatures lower than 10°C or when leaf wetness lasted <24 h. At 48-h leaf wetness, disease incidence was 0, 28, 77, and 85% at 10, 15, 20, and 25°C, respectively. Middle and lower leaves on the plant were more susceptible than upper leaves. In microscopic observations of inoculated leaves, >50% of conidia germinated at temperatures >15°C after 24-h leaf wetness. More appressoria formed at higher temperatures (15–25°C) with extended duration of leaf wetness (24–72 h). At 48-h leaf wetness, appressorium formation was 0, 8, 26, and 73% at 10, 15, 20, and 25°C, respectively. These results suggest that temperature and leaf wetness duration were important factors for infection of gentian leaves.     

Responses of Erysiphe graminis germlings to contact with artificial and host surfaces

Germling development by Erysiphe graminis f. sp. hordei was compared between conidia held in a simulated air-borne state on microthreads constructed from safety-line threads produced by orb--weaving spiders (Araneus diadematus), and conidia inoculated onto glass, agar, or living or dead barley coleoptile epidermes. Suspended conidia germinated but generally produced only multiple short germ tubes. Conidia on living or dead coleoptiles, bathed from beneath with 0.01 Ca(NO₃)₂ solution, generally produced one short germ tube and a second germ tube which elongated and formed a normal appressorium. On glass and agar, multiple short germ tubes were sometimes formed but long germ tubes were formed less frequently than on host epidermis. When conidia with short germ tubes were transferred from microthreads to coleoptiles, they produced a long germ tube which differentiated an appressorium. Conidia with a single short germ tube were also transferred from microthreads so that only the tip of the short germ tube was in contact with a leaf epidermal strip layed on agar, whilst the conidium rested on the agar. Long germ tubes were formed more frequently by such conidia than by controls which had no contact with the leaf epidermis. This suggested that a stimulus causing elongation of the second tube was perceived through the short germ tube in contact with the epidermal strip. Where long germ tubes made contact with the epidermal strip, normal appressoria were formed more frequently than where the long tube made contact with the agar surface alone. The results indicate that germlings develop through distinct stages in response to particular stimuli.     

Calcium/calmodulin-dependent signaling for prepenetration development in <Emphasis Type="Italic">Cochliobolus miyabeanus</Emphasis> infecting rice

Cochliobolus miyabeanus forms a specialized infection structure, an appressorium, to infect rice. Contacting a hard surface induces appressorium formation in C. miyabeanus, while the hydrophobicity of the substratum does not affect this morphogenic infection event. To determine whether the calcium/calmodulin-dependent signaling system is involved in prepenetration morphogenesis in C. miyabeanus, the effects of a calcium chelator (ethylene glycol tetraacetic acid; EGTA), phospholipase C inhibitor (neomycin), intracellular calcium channel blocker (TMB-8), calmodulin antagonists (chlorpromazine, phenoxybenzamine, and W-7), and calcineurin inhibitor (cyclosporin A) on morphogenesis and infection were examined. Addition of Ca²⁺ and the calcium ionophore A23187 did not affect conidial germination, while the number of appressoria decreased with higher concentrations. EGTA inhibited conidial germination and appressorium formation. The calcium channel blocker did not affect appressorium formation at any concentration; however, calmodulin antagonists and the calcineurin inhibitor specifically reduced appressorium formation at the micromolar level. One of the calmodulin antagonists, W-7, also inhibited accumulation of mRNA of the calmodulin gene within germinating conidia and/or appressorium-forming germ tubes. Thus, biochemical processes controlled by the calcium/calmodulin signaling system seem to be involved in the induction of prepenetration morphogenesis on rice.     

The early development of Erysiphe pisi on Pisum sativum L.

Erysiphe pisi , the powdery mildew pathogen of Pisum sativum , followed a developmental sequence that allowed the identification of 10 distinct growth stages (GS) over 30 h following inoculation. The growth stages were ungerminated conidia (GS1), germinated conidia, having produced a germ tube (GS2), germlings where the germ tube had forked (GS3), germlings with a multi-lobed germ tube (GS4), germlings with a single hypha (GS5), germlings with two (GS6 and 7) or three (GS8 and 9) hyphae, one of which may have formed from the appressorium (GS7 and 9), and germlings with abnormally long germ tubes (GS10), which did not develop hyphae. Conidia germinated rapidly, with a quarter of conidia producing germ tubes by 2 h after inoculation (hai). Most germlings produced multi-lobed appressoria, which showed considerable variation in structure. Haustoria, although often difficult to visualize, were first seen 4 hai, and the first hyphae 14 hai, growing from the body of the conidium. Subsequent hyphae developed from both the body of the conidium and from the appressorium.     

Effect of pentachloronitrobenzene,pentachloroaniline, and albinism on epidermal penetration by appressoria of Pyricularia

Pentachloroaniline or pentachloronitrobenzene at concentrations between 0.5 and 5 μg/ml blocked appressorial melanization and appressorial penetration of onion epidermal cell walls by Pyricularia oryzae, strain P-2, without affecting conidial germination or appressorial formation. Among polyphenol oxidase inhibitors, salicylhydroxamic acid, and phenylthiourea at 100 μg/ml inhibited appressorial penetration of onion epidermis without affecting appressorial melanization, whereas sodium diethyldithiocarbamate and 2,3-naphthalenediol did not affect either process at 1.0 μg/ml but inhibited conidial germination and appressorial formation at 10 μg/ml. EDTA was without effect on either melanization or penetration at 100 μg/ml. The melanized appressoria of Pyricularia grisea, strain BL-3, penetrated onion epidermal cell walls, whereas nonpigmented appressoria of an albino mutant (AL-3) of this fungus did not. Melanization and penetration ability of AL-3 appressoria were restored by adding melanin biosynthesis intermediates scytalone or 1,8-dihydroxynaphthalene (1,8-DHN). Tricyclazole blocked melanization and epidermal penetration by AL-3 appressoria treated with scytalone but not by those treated with 1,8-DHN. Scytalone (20 μg/ml) increased the effectiveness of tricyclazole in blocking penetration by appressoria of BL-3. Tricyclazole was about 10 times more effective in inhibiting penetration by P-2 appressoria than by BL-3 appressoria.     

Subcuticular-Intracellular Hemibiotrophic and Intercellular Necrotrophic Development of Colletotrichum acutatum on Almond

ABSTRACT The early infection and colonization processes of Colletotrichum acutatum on leaves and petals of two almond cultivars with different susceptibility to anthracnose (i.e., cvs. Carmel and Nonpareil) were examined using digital image analysis of light micrographs and histological techniques. Inoculated tissue surfaces were evaluated at selected times after inoculation and incubation at 20 degrees C. Depth maps and line profiles of the digital image analysis allowed rapid depth quantification of fungal colonization in numerous tissue samples. The results showed that the early development of C. acutatum on petals was different from that on leaf tissue. On petals, conidia germinated more rapidly, germ tubes were longer, and fewer appressoria developed than on leaves. On both tissues, penetration by the pathogen occurred from appressoria and host colonization was first subcuticular and then intracellular. On petals, colonizing hyphae were first observed 24 h after inoculation and incubation at 20 degrees C, whereas on leaves they were seen 48 to 72 h after inoculation. Intercellular hyphae were formed before host cells became necrotic and macroscopic lesions developed on petals >/=48 h and on leaves >/=96 h after inoculation. Histological studies complemented data obtained by digital image analysis and showed that the fungus produced infection vesicles and broad hyphae below the cuticle and in epidermal cells. In both tissues, during the first 24 to 48 h after penetration fungal colonization was biotrophic based on the presence of healthy host cells adjacent to fungal hyphae. Later, during intercellular growth, the host-pathogen interaction became necrotrophic with collapsed host cells. Quantitative differences in appressorium formation and host colonization were found between the two almond cultivars studied. Thus, on the less susceptible cv. Nonpareil fewer appressoria developed and host colonization was reduced compared with that on cv. Carmel.     

Melanin biosynthesis as a prerequisite for penetration by appressoria of Colletotrichum lagenarium: Site of inhibition by melanin-inhibiting fungicides and their action on appressoria

Melanin biosynthesis by appressoria was studied in relation to their penetrating ability using tricyclazole [5-methyl-1,2,4-triazolo(3,4-b)benzothiazole], pp 389 [4,5-dihydro-4-methyltetra-zolo(1,5-a)quinazolin-5-one], and pyroquilon [1,2,5,6-tetrahydropyrrolo(3,2,1-i,j)quinolin-4-one], and color mutants of Colletotrichum lagenarium. Tricyclazole at 100 μM inhibited melanin biosynthesis by appressoria of C. lagenairum 104-T, and caused accumulation of 3,4-dihydro-4,8-dihydroxy-1(2H)naphthalenone (DDN) in the culture medium. By contrast, DDN was not detected in culture media of tricyclazole-treated mutant 8015, which is defective in the enzyme involved in the conversion of scytalone to 1,3,8-trihydroxynaphthalene (1,3,8-THN). Vermelone restored melanization of appressoria of albino mutant 79215 and of the parent strain 104-T treated with tricyclazole, pp 389, and pyroquilon; however, scytalone restored melanization only in appressoria of albino mutant 79215. These results indicate that tricyclazole, pp 389, and pyroquilon inhibit the conversion of 1,3,8-THN to vermelone in the melanin biosynthetic pathway of appressoria of C. lagenarium. Colorless appressoria formed in the presence of the three melanin-inhibiting chemicals germinated laterally on nitrocellulose membranes and rarely penetrated the membranes. On the other hand, when pigmented appressoria were restored by application of vermelone in the presence of the three chemicals, lateral germination of the appressoria was largely suppressed, and the membranes were effectively penetrated. From these results, it is concluded that the major effect of tricyclazole, pp 389, and pyroquilon on appressoria of C. lagenarium, causing failure of penetration, is the inhibition of melanization. Effects of the chemicals on other metabolic functions can be precluded as significant factors affecting the penetration process.     

Strawberry Anthracnose: Histopathology of Colletotrichum acutatum and C. fragariae

ABSTRACT Ontogeny of the invasion process by Colletotrichum acutatum and C. fragariae was studied on petioles and stolons of the strawberry cultivar Chandler using light and electron microscopy. The invasion of host tissue by each fungal species was similar; however, each invasion event occurred more rapidly with C. fragariae than with C. acutatum. Following cuticular penetration via an appressorium, subsequent steps of invasion involved hyphal growth within the cuticle and within the cell walls of epidermal, subepidermal, and subtending cells. Both species of fungi began invasion with a brief biotrophic phase before entering an extended necrotrophic phase. Acervuli formed once the cortical tissue had been moderately disrupted and began with the development of a stroma just beneath the outer periclinal epidermal walls. Acervuli erupted through the cuticle and released conidia. Invasion of the vascular tissue typically occurred after acervulus maturation and remained minimal. Chitin distribution in walls of C. fragariae was visualized with gold-labeled wheat germ agglutinin. The outer layer of bilayered walls of conidia, germ tubes, and appressoria contained less chitin than unilayered hyphae in planta.     

Extracellular matrix of <Emphasis Type="Italic">Magnaporthe oryzae</Emphasis> may have a role in host adhesion during fungal penetration and is digested by matrix metalloproteinases

Spores and infection structures such as germ tubes and appressoria of Magnaporthe oryzae, the fungus causing blast disease of wheat, produced an extracellular matrix (ECM) on the surfaces of host leaves during fungal differentiation. The chemical components and function of the ECM were studied to understand the pathological roles using two immunological techniques and ECM-digesting enzymes. The ECM was characterized by fibrous and amorphous materials, located in the spaces between fungal cell walls and plant cuticles. Immunohistochemical and immunoelectron microscopy suggested that ECM includes components positively reacted with antibodies of four animal cell adhesion factors (collagen VI, vitronectin, fibronectin and laminin) and an animal integrin α3. ECM, incubated on a cellulose membrane, was rapidly digested by matrix metalloproteinases (collagenase and gelatinase B), resulting in the detachment of most infection structures from membrane surfaces. Both ultrastructural observation and immunological responses showed that more ECM was located at the appressoria than at the spores and germ tubes. This result suggested that appressoria needed a powerful adhesion force for aggressive action of penetration pegs into plant cuticles. An erratum to this article can be found at     

Influence of Nutrient and Environmental Factors on Conidial Germination of Potebniamyces pyri

ABSTRACT Potebniamyces pyri is the causal agent of Phacidiopycnis rot, a postharvest disease of pears. Infection of fruit occurs in the orchard, and symptoms develop during storage. Conidial germination of P. pyri in response to nutrient, temperature, wetness duration, relative humidity (RH), and pH was determined in vitro. Conidia germinated by either budding or developing germ tubes in various concentrations of pear juice solutions. The mode of conidial germination was nutrient-dependent. Low nutrient levels favored budding, whereas high nutrient levels favored germ tube development. Conidia germinated at 0 to 30 degrees C but not at 35 degrees C, with optimum temperature between 20 and 25 degrees C. Wetness durations of 4 to 5 h and 6 to 8 h at optimum temperature were required for budding and developing germ tubes, respectively, and 20 to 24 h of wetness was required to reach germination peaks. Regardless of temperature, conidia germinated primarily by budding in 10% pear juice. Secondary conidia, produced by budding of conidia, initially increased their dimensions and later germinated at 0 to 25 degrees C in the same manner as mother conidia. No germination of secondary conidia occurred at 30 degrees C. Germ tubes from conidia elongated at 0 to 25 degrees C but not at 30 degrees C. No germination occurred at 相似文献   

Effect of humidity and temperature on conidial germination and appressorium development of two Philippine isolates of the mango anthracnose pathogen Colletotrichum gloeosporioides

A comparison of rates of germination and appressorium formation by an isolate of Colletotrichum gloeosporioides on mango leaves, fruit surfaces and cellophane membranes showed that behaviour was broadly similar on all three substrates. Frequency of appressorium formation was slightly higher on cellophane membranes, and both hyaline and melanized appressoria were formed. Only melanized appressoria were formed on mango surfaces. In vitro experiments on membranes showed comparative differences in physiological behaviour with temperature for two Philippine isolates of C. gloeosporioides . The most stimulatory temperature for production of appressoria differed in isolates I-2 and I-4 (25 and 20°C, respectively). At 30°C more appressoria became melanized than at lower temperatures, but the frequency of formation of penetration pegs was highest at 25°C. Conidia of C. gloeosporioides germinated on cellophane membranes at relative humidities as low as 95%, but the percentage of conidia germinating and forming appressoria increased as the RH approached 100%. Approximately 18% of conidia of C. gloeosporioides I-2 held at 62 and 86% RH for 4 weeks retained viability, and some were capable of forming appressoria when placed at 100% RH. These results have implications for epidemiological models for disease control.     

Microscopical studies of the infection of gerbara flowers byBotrytis cinerea

The formation of lesions on ray florets of gerbera flowers caused by single conidia ofBotrytis cinerea was studied in two cultivars infected by two isolates of the pathogen. No differences in reaction after inoculation with conidia of either isolate were seen on either cultivar. The conidia produced usually one germ tube not longer than 10 m, but conidia with five germ tubes were also seen. Direct penetration of germ tubes through the upper cuticle of ray florets was observed. No appressoria or other specialised structures were observed before penetration, and degradation of the cuticle did not occur. Germination of conidia and subsequent flower infection was dependent on the availability of free water, but not on the addition of external nutrients.Between 18 to 25°C, fungal development usually stopped after cuticle penetration, two to four cells around the site of penetration becoming necrotic. This number did not increase when inoculated flowers were subsequently placed at 4°C, conditions conductive for the formation of spreading lesions. When flowers were incubated constantly at 4°C, lesions became visible 3 days after inoculation as a group of 10 to 14 cells. Initially from a vesicle-like structure, mycelium spread subcuticularly or in the lumen of epidermal cells resulting in the death of 40 to 50 cells at 18 days after inoculation. Ungerminated conidia and conidial germlings which has not yet penetrated the cuticle did not cause any visible symptoms in underlying epidermal cells.     

Comparative colonisation by virulent versus avirulent <Emphasis Type="Italic">Pyricularia oryzae</Emphasis> on wild <Emphasis Type="Italic">Oryza australiensis</Emphasis>

Pyricularia oryzae (rice blast) conidial development at pre-penetration stage determines success or otherwise of infection inside the rice host plants. Studies on conidial germination and growth on the leaf surface in commercial rice (Oryza sativa) report differently, dependent upon host type and level of blast resistance. Although wild rice (O. australiensis) is known to be an alternative host of blast, the interaction between P. oryzae conidia and wild O. australiensis on its leaf surface has not been previously studied. We found significant (P?<?0.001) differences in conidial development between two blast isolates with different virulence in terms of conidial germination, germ tube growth and appressoria formation on both wild and cultivated rice. Conidial germination at 6 h post-inoculation (hpi) for the virulent isolate was significantly (P?<?0.001) delayed. Germ tubes of the avirulent isolate conidia grew significantly (P?<?0.001) faster and with significantly (P?<?0.001) longer germ tubes than from virulent conidia. Appressoria development for the virulent isolate was significantly (P?<?0.001) faster at its later growth stages of 12 and 18 hpi when approximately 100% of germ tubes formed appressoria. In contrast, formation rate of appressoria for the avirulent isolate was significantly (P?<?0.001) slower and only reached 76% of germ tubes forming appressoria. Appressoria formation on O. australiensis was significantly (P?<?0.001) greater than the formation on O. sativa for both virulent and avirulent P. oryzae at 12 hpi, a clear indication that host type influences the extent of appressoria formation.