Effects of an Interaction Between Inoculum Density and Temperature on Germination of Puccinia allii Urediniospores and Leek Rust Progress

ABSTRACT Controlled environment experiments were conducted to study the effects of inoculum density, temperature, and their interaction on germination of Puccinia allii urediniospores and infection of leek leaves. Percent germination of P. allii urediniospores and percent branching of germ tubes increased with 3 density of urediniospores and approached a plateau for densities above approximately 20 spores cm(-2) of leaf area. Percent germination was highest at 12 to 21 degrees C, a wide-range temperature optimum. Branching occurred at temperatures ranging from 5 to 25 degrees C, but there were few germ tubes branching at 25 degrees C. P. allii successfully infected leek leaves at temperatures ranging from 7 to 22 degrees C. The number of pustules produced increased with urediniospore density on leek leaves. At low spore densities, pustule production was little affected by temperature; at higher spore densities, pustule production was greatest between 9 to 11 degrees C, and numbers of pustules decreased greatly with temperature increasing above this optimum. Latent period was affected by temperature, with latent period being shortet between 19 and 22 degrees C, and latent period increasing when temperature decreased. Latent periods became approximately 1.8 days shorter for every 10-fold increase in spore density. The rate of pustule production increased with increasing spore density on leaves and was greatest between 11 to 14 degrees C. Computer simulation of leek rust progress based on the found relationships suggested that at optimal temperatures the development of leek rust epidemics may be little affected by initial spore density and density caused by each pustule, but that at sub- and supra-optimal temperatures the development is greatly affected by these variables.     

Effect of solar radiation on severity of soybean rust

Soybean rust (SBR), caused by Phakopsora pachyrhizi, is a damaging fungal disease of soybean (Glycine max). Although solar radiation can reduce SBR urediniospore survival, limited information is available on how solar radiation affects SBR progress within soybean canopies. Such information can aid in developing accurate SBR prediction models. To manipulate light penetration into soybean canopies, structures of shade cloth attenuating 30, 40, and 60% sunlight were constructed over soybean plots. In each plot, weekly evaluations of severity in lower, middle, and upper canopies, and daily temperature and relative humidity were recorded. Final plant height and leaf area index were also recorded for each plot. The correlation between amount of epicuticular wax and susceptibility of leaves in the lower, middle, and upper canopies was assessed with a detached leaf assay. Final disease severity was 46 to 150% greater in the lower canopy of all plots and in the middle canopy of 40 and 60% shaded plots. While daytime temperature within the canopy of nonshaded soybean was greater than shaded soybean by 2 to 3°C, temperatures recorded throughout typical evenings and mornings of the growing season in all treatments were within the range (10 to 28.5°C) for SBR development as was relative humidity. This indicates temperature and relative humidity were not limiting factors in this experiment. Epicuticular wax and disease severity in detached leaf assays from the upper canopy had significant negative correlation (P = 0.009, R = -0.84) regardless of shade treatment. In laboratory experiments, increasing simulated total solar radiation (UVA, UVB, and PAR) from 0.15 to 11.66 MJ m(-2) increased mortality of urediniospores from 2 to 91%. Variability in disease development across canopy heights in early planted soybean may be attributed to the effects of solar radiation not only on urediniospore viability, but also on plant height, leaf area index, and epicuticular wax, which influence disease development of SBR. These results provide an understanding of the effect solar radiation has on the progression of SBR within the soybean canopy.     

Extended survival of Puccinia graminis f. sp. tritici urediniospores: implications for biosecurity and on‐farm management

Puccinia graminis f. sp. tritici (Pgt), the causal organism of stem rust, is of global importance across wheat‐growing countries. However, some epidemics commence without the obvious presence of ‘alternate’ or ‘green bridge’ hosts, suggesting urediniospores can survive in the absence of suitable host plants for many weeks. Testing a range of inert material types, including metals, plastics, fabrics and woods, highlighted a significant effect of material type and temperature on urediniospore viability (P < 0.001), with urediniospores remaining attached and viable on these materials (aluminium, paper, rubber, all fabric and all woods) for up to 365 days at 23/8 °C day/night. At 36/14 °C day/night, urediniospore viability was retained for a maximum of 300 days on denim and jute. Furthermore, at 45/15 °C day/night, urediniospores remained viable for a maximum of 180 days on cotton and jute. The frequency of recovery of attached urediniospores was also dependent upon the material type, with significant differences between materials in their abilities to retain urediniospores after washing (P < 0.001). Urediniospores recovered even after 300 or 365 days from the lower two temperature regimes successfully initiated infections of wheat seedlings. Results confirm the potential importance of inert materials as long‐term carriers of viable Pgt urediniospores, highlighting risks of spread of new pathotypes and strains across wheat‐growing regions, the significant biosecurity implications for contaminated carrier materials, and its likely survival across seasons without a host.     

Effects of temperature on urediniospore germination, germ tube growth, and initiation of infection in soybean by phakopsora isolates

ABSTRACT Temperature is a critical factor in plant disease development. As part of a research program to determine how specific environmental variables affect soybean rust, we determined temperature effects on urediniospore germination and germ tube growth of four isolates of Phakopsora pachyrhizi, one each from Brazil, Hawaii, Taiwan, and Zimbabwe, and an isolate of P. meibomiae from Puerto Rico, collected over a 25-year period. Also compared were the effects of temperature during a night dew period on initiation of disease by the P. pachyrhizi isolates. All variables were fit to a nonlinear beta function with temperature as the independent variable. Minimum, maximum, and optimum temperatures, along with shape parameters of the beta function for each variable, were statistically analyzed. All Phakopsora isolates behaved similarly as to how temperature affected urediniospore germination, germ tube growth, and initiation of disease. The results suggest that P. pachyrhizi has changed little in the past few decades with respect to how it responds to temperature and that previously collected research data continues to be valid, simplifying the development of soybean rust disease models.     

Quantitative aspects of the spread of asian soybean rust in the southeastern United States, 2005 to 2006

ABSTRACT The regional dynamics of soybean rust, caused by Phakopsora pachyrhizi, in six southeastern states (Florida, Georgia, Alabama, South Carolina, North Carolina, and Virginia) in 2005 and 2006 were analyzed based on disease records collected as part of U.S. Department of Agriculture's soybean rust surveillance and monitoring program. The season-long rate of temporal disease progress averaged approximately 0.5 new cases day(1) and was higher in nonsentinel soybean (Glycine max) plots than in sentinel soybean plots and kudzu (Pueraria lobata) plots. Despite the early detection of rust on kudzu in January and/or February each year (representing the final phase of the previous year's epidemic), the disease developed slowly during the spring and early summer on this host species and did not enter its exponential phase until late August, more than 1 month after it did so on soybean. On soybean, cases occurred very sporadically before the beginning of July, after which their number increased rapidly. Thus, while kudzu likely provides the initial inoculum for epidemics on soybean, the rapid increase in disease prevalence on kudzu toward the end of the season appears to be driven by inoculum produced on soybean. Of 112 soybean cases with growth stage data, only one occurred during vegetative crop development while approximately 75% occurred at stage R6 (full seed) or higher. The median nearest-neighbor distance of spread among cases was approximately 70 km in both years, with 10% of the distances each being below approximately 30 km and above approximately 200 km. Considering only the epidemic on soybean, the disease expanded at an average rate of 8.8 and 10.4 km day(1) in 2005 and 2006, respectively. These rates are at the lower range of those reported for the annual spread of tobacco blue mold from the Caribbean Basin through the southeastern United States. Regional spread of soybean rust may be limited by the slow disease progress on kudzu during the first half of the year combined with the short period available for disease establishment on soybean during the vulnerable phase of host reproductive development, although low inoculum availability in 2005 and dry conditions in 2006 also may have reduced epidemic potential.     

Effects of temperature and humidity on the survival of urediniospores of gladiolus rust (<Emphasis Type="Italic">Uromyces transversalis</Emphasis>)

Uromyces transversalis is an autoecious microcyclic rust mainly infecting Gladiolus spp. The pathogen is considered of plant quarantine importance in Europe and the USA. In 2006, the pathogen was found for the first time in the USA in several commercial nurseries in Florida and California. The US Department of Agriculture (USDA) initiated an eradication programme that recommended the immediate removal and destruction of infected plants followed by a host-free period, use of a fungicide treatment schedule, and equipment decontamination. In support of this plan, a study was conducted to determine how long urediniospores of U. transversalis would continue to germinate at temperatures of 2.8, 15.0, 18.8 and 25.0°C under controlled relative humidities (RH) of 11, 23, 43, 75, 93 and 100%. Choice of temperature and humidity parameters were mostly based on historical multi-year climate data from areas where the disease was detected in California and Florida. Analysis of variance (ANOVA) indicated no significant effect of RH on urediniospore germination but a highly significant effect of temperature. No germinating urediniospores were detected after 79 days for any treatment, but the 15°C treatment was more likely to be the result of germination independent of any low or high temperature-induced spore quiescence. Thus, lack of germination after 79 days was probably a good indicator of the lack of viable spores after this time for the 15°C treatment.     

小麦锈菌夏孢子经盐酸处理后的形态观察

为了快速区分小麦条锈菌、叶锈菌和秆锈菌,用不同浓度的盐酸处理3种小麦锈菌的新鲜夏孢子和在4 ℃冰箱内保存4个月以上的锈菌夏孢子,在显微镜下观察夏孢子形态以及原生质体变化。结果表明, 24.5%、28.5%、32.5%、36.5%4种浓度的盐酸处理条锈菌夏孢子后其原生质体均浓缩成多个分散的小团。而同样方法处理叶锈菌和秆锈菌新鲜菌种时其原生质体浓缩成一个大团或多个小团,浓缩成一个大团的比例随盐酸浓度的增大而提高;用不同浓度盐酸处理保存4个月以上的2种小麦锈菌时,95%以上的夏孢子原生质体浓缩成多个分散的小团,少数（不超过5%）的夏孢子原生质体浓缩成一个大团。研究还发现叶锈菌和秆锈菌夏孢子活力、盐酸浓度等对夏孢子原生质体浓缩状况有很大影响,且与病菌生理小种有一定的关系。因此,锈菌夏孢子经36.5%浓盐酸处理后的原生质体浓缩状况只能作为小麦锈病田间快速诊断检测的辅助手段,不能作为锈病种类鉴别的唯一标准,必须结合病害症状特征、孢子形态以及分子生物学方法等进行综合判别。     

Evaluation of morphology of infection structures in distinguishing between different Allium rust fungi

Fifteen isolates of rust fungi were collected in the United Kingdom and in the Netherlands from severalAllium species. The samples represented three putative rust species. The morphology of the telia, teliospores and urediniospores was investigated. From urediniospores infection structures were induced on leek, and their morphology was described. Telia and teliospores were not available on every sample. Morphology of the infection structures clearly differentiated between ‘leek’ type and ‘chive’ type isolates. The morphology of infection structures ofPuccinia mixta is very similar to that ofUromyces ambiguus, but clearly distinct fromPuccinia allii sensu Jennings from leek. Quantitative differences in urediniospore characters differentiated between the putative species, but there was overlap between the taxa. We conclude that morphology of infection structures of urediniosporelings is a useful trait for identification of rust isolates fromAllium. This is especially true where more than one rust species may occur on the same host species, as withAllium fistulosum.     

First release of a fungal classical biocontrol agent against an invasive alien weed in Europe: biology of the rust,Puccinia komarovii var. glanduliferae

The rust fungus Puccinia komarovii var. glanduliferae was first identified infecting Impatiens glandulifera in its native range (western Himalayas) between 2006 and 2010. Subsequently, it was imported into quarantine in the UK for evaluation as a classical biocontrol agent. To assess the safety of the rust, plant species relevant to Europe were tested for susceptibility. To confirm the life cycle, all infective spore stages were inoculated on I. glandulifera to follow disease progression. Teliospores were primed using bleaching and low temperatures to break dormancy. Temperature and dew period experiments using urediniospores were conducted to assess the parameters required for infection. Of the 74 plant species tested, only I. balsamina, an ornamental species, was fully susceptible to urediniospore inoculum. The life cycle of the rust – an autoecious, full‐cycled species with five spore stages – was confirmed. Urediniospores were infective between 5 and 25°C, with an optimum at 15°C. A minimum of 8 h dew period was required to achieve consistent infection. Based on a pest risk assessment, the rust poses no threat to native biodiversity within EU Member States; making P. komarovii var. glanduliferae a suitable candidate as the first fungal classical biocontrol agent against an exotic weed in the region.     

Glyphosate reduces urediniospore development and Puccinia psidii disease severity on Eucalyptus grandis

BACKGROUND: Recent studies have shown the effects of glyphosate drift on decreasing rust intensity on Eucalyptus grandis plants. However, the effects of the herbicide on Puccinia psidii initial development are unknown. In this study the systemic action of glyphosate on rust severity was evaluated on Eucalyptus plants maintained under greenhouse conditions. Urediniospore germination and apressorium formation on detached leaves and on water agar medium, previously treated with glyphosate, were also evaluated. RESULTS: Rust severity and the number of urediniospores per leaf area were significantly reduced with increasing glyphosate doses, even on branches not directly treated with the herbicide, indicating a systemic effect of glyphosate on pathogen development. Similarly, higher glyphosate doses also reduced germination and apressorium formation on detached Eucalyptus leaves, regardless of the direct application of the product on the leaf limb or on the petiole base. Puccinia psidii urediniospore germination in water agar medium also decreased with increasing herbicide doses. CONCLUSIONS: Reductions in germination and apressorium formation of P. psidii urediniospores with increasing glyphosate dose indicate that a lower severity and intensity of the disease may perhaps be due to blockage of the shikimic acid pathway in the fungal metabolic system. Copyright © 2011 Society of Chemical Industry     

Relating epidemic progress from a general disease model to seasonal appearance time of rusts in the United States: implications for soybean rust

ABSTRACT Soybean rust, Phakopsora pachyrhizi, has been considered a threat to the production of the U.S. soybean, Glycine max. During the past decade, this disease gradually spread to Africa, South America, and recently to the United States. Previous soybean rust risk assessments with an assumption of availability of spores early in a season showed that weather conditions (dew and temperature) during a growing season, in general, are suitable for disease development in U.S. soybean-growing regions. Predicting the time of rust appearance in a field is critical to determining the destructive potential of rusts, including soybean rust. In this study, comparative epidemiology was used to assess likely rust incipient time in four locations within the U.S. Soybean Belt from south to north: Baton Rouge, LA; Charlotte, NC; Indianapolis, IN; and Minneapolis, MN. Temperature effects on the infection cycle of five rusts occurring in the Midwest were evaluated using a general disease model. The likely incipient times were examined with the modeling results. Among the rusts studied, early-appearing rusts had suitable conditions for development earlier in a season. However, a lag period of several weeks to more than 3 months was found from the time when conditions are suitable for a rust to develop or when hosts are available to the time when the rust was detected in fields. Length of the lag period differed among the rust species examined. If nature of long-distance dispersal is not significantly different among the rusts, implications of our study to the expected seasonal soybean rust incipience in fields lead to two possible scenarios: (i) average appearance time of soybean rust across the Soybean Belt should be somewhere between appearance times of common corn rust and southern corn rust, and (ii) with late appearance of the disease, late-planted soybean in the south has greater risk.     

Plantainlily rust fungus is distinct from daylily rust fungus

Plantainlily rust fungus, Puccinia funkiae, was proven to host-alternate between Hosta species (plantainlilies) and a patrinia, Patrinia villosa. Unlike Puccinia hemerocallidis (daylily rust fungus), P. funkiae was not pathogenic on Patrinia scabiosaefolia or on Hemerocallis species (daylilies). The urediniospores were also larger and thicker-walled than those of the daylily rust fungus. The two or three urediniospore germ pores of the plantainlily rust fungus were equatorially distributed, whereas there were several germ pores in P. hemerocallidis spores, and they were scattered over the wall. Thus, P. funkiae is biologically and taxonomically distinct from P. hemerocallidis.     

Effect of host genotype on leaf rust (Puccinia triticina) lesion development and urediniospore production in wheat seedlings

Experiments were performed in controlled conditions to compare several aggressiveness components of an isolate of Puccinia triticina on different wheat ( Triticum aestivum ) genotypes. Latency period, urediniospore production per lesion and per unit of sporulating surface, lesion size, and nitrogen and carbon content of the spores were measured on four host genotypes. Three of the host cultivars (Soissons, Altria and Isengrain) were susceptible and the fourth (Trémie) was resistant to the isolate used in the experiments, producing a mesothetic infection type. In all the analyses, lesion density was used as a covariable. The latent period was identical on the susceptible host cultivars, and there were no marked differences in the urediniospore production capacity when related to the sporulating surface area. In cv. Isengrain, differences in lesion size relative to the other susceptible cultivars resulted in lower urediniospore production per lesion. Urediniospore production per lesion and lesion size were density dependent and decreased exponentially with increasing lesion density. Urediniospore production per unit of sporulating surface was either independent of lesion density or marginally dependent (decreased linearly with a limited rate). On the resistant cultivar, Trémie, the latent period was longer and of a much greater variance compared with the susceptible cultivars. Urediniospore production per lesion was considerably reduced, but this reduction was fully accounted for by a reduction in lesion size, the urediniospore production capacity of the diseased tissue remaining equivalent to that observed in the susceptible cultivars. In all cultivars the C and N contents of the urediniospores were considered unchanged, even if minor differences were detected.     

小豆锈病病原菌鉴定

 小豆锈病是小豆的重要病害。然而,小豆锈病病原菌在我国还没有被鉴定。本文对从黑龙江省和内蒙古自治区采集的5个小豆锈病菌样品的分类地位进行了研究。形态学观察表明,小豆锈病菌夏孢子芽孔位于赤道线或赤道线上方,冬孢子平均壁厚为2.0 μm。分子标记检测发现,小豆锈病菌不能被疣顶单胞锈菌特异性引物对UA-ITSF/UA-ITSR扩增,小豆锈病菌和豇豆锈病菌不能被豇豆单胞锈特异性引物对UV-ITSF/UV-ITSR区分。对豇豆单胞锈特异性引物对扩增的4个小豆锈病菌样品PCR产物进行测序,比对分析表明目标序列与小豆单胞锈ITS序列的同源性为99%~100%。基于夏孢子的芽孔位置、冬孢子壁的厚度、疣顶单胞锈菌及豇豆单胞锈特异性引物检测结果和ITS序列分析,小豆锈病菌被鉴定为小豆单胞锈。     

Day and night temperature effects on vegetative growth of Erodium Cicutarium

The vegetative growth response of Erodium cicutarium (L.) ?Her. ex Ait. to various day:night temperature regimes was studied under controlled environment conditions. Dry matter production was greatest with day temperatures of 18 to 34^C combined with night temperatures of 12 to 18^C. A high night temperature of 24^C was very detrimental, reducing dry matter production to 15 to 25% of that attained at 12^C. The optimum mean daily temperature for growth of E. cicutarium is predicted to be in the range 17 to 20^C. Little growth is predicted at mean daily temperatures below 5^C and above 30^C. Partitioning of biomass in leaves, stems and roots was markedly affected by day and night temperatures. Stem weight ratio was greatest at day temperatures of 18 to 34^C and night temperatures of 18 to 24^C. Maximum leaf weight ratio occurred at day and night temperatures of 10 to 18^C. Root biomass was little affected by day temperatures, but was greatest at a night temperature of 12^C, declining substantially as night temperature increased from 12 to 24^C. Results are discussed in terms of the potential for E. cicutarium to become an increasingly troublesome weed in crops of the Canadian prairies and in terms of possible management strategies for its control.     

Molecular diagnosis of Puccinia psidii (guava rust) – a quarantine threat to Australian eucalypt and Myrtaceae biodiversity

A species-specific, nested polymerase chain reaction (PCR)-based detection assay, using two primer sets designed from the rRNA ITS region, was developed for Puccinia psidii . Detection sensitivities of one urediniospore alone, or one or two urediniospores in the presence of pollen or leaf tissues, respectively, were observed. The assay reliably, accurately and sensitively detected the rust from naturally infected, geographically widespread eucalypt and fruit tree plantation and nursery species from diverse tissues types (e.g. leaves, flowers, fruits, pollen, seeds and woody material) including symptomless or cryptically contaminated plants or plant tissue. Independent testing in Brazil and Australia demonstrated the international inter-laboratory transferability of the P. psidii assay required for germplasm screening, disease monitoring and quarantine and incursion management, towards which the assay has already been employed.     

Inter-population variabilities in seed mass and germination of Panicum turgidum and Pennisetum divisum in the desert of Kuwait

Understanding variability in seed germination among populations is essential for planning an effective germplasm collection for restoration and conservation purposes.The knowledge of germination and dormancy patterns among populations of desert grasses is crucial for determining the potential of the species and populations to be used for restoration and conservation as well as forage production.Variability in seed germination of Panicum turgidum Forssk and Pennisetum divisum(Gmel.)Henr.in the desert of Kuwait was evaluated in different populations in May 2017.Experiment of seed germination(25 seeds and 4 replicates)was conducted for each population at night/day temperatures of 15℃/20℃and 20℃/30℃under the following light condition:continuous darkness or 12 h/12 h light/dark.Results showed that seed masses of both species strongly varied according to their seed provenances,and both species produced heavier seeds in population with a higher soil electrical conductivity.Seed germination percentage considerably varied between two species,and the variation in P.turgidum was greater(17%–49%)than that of P.divisum(72%–93%).Germination percentage in P.turgidum was greater at high temperature(20℃/30℃)than at low temperature(15℃/20℃).However,temperature regimes had no effect on germination percentage of P.divisum seeds.Mean germination time of both species exhibited significant inter-population variability.This result is especially relevant to assure the selection of the best population of each species and the regeneration success of the species.Besides this,inter-population variability also provides valuable information for enhancing our understanding of the mechanisms that regulate seed germination and how they might be related to seed provenance.     

Temperature adaptation in Australasian populations of Puccinia striiformis f. sp. tritici

Stripe rust, caused by Puccinia striiformis f. sp. tritici, is one of the major fungal pathogens of wheat. A new pathotype was introduced to Australia in 2002 and several derivative pathotypes were detected in subsequent seasons. It has been suggested that the severity of stripe rust outbreaks in Australia since 2002 could be as a result of traits other than virulence in the pathogen population. This study was conducted to investigate the hypothesis that the stripe rust pathogen population dominant in Australia since 2002 was better adapted to warm temperature conditions compared to previous pathogen populations. Sixteen pathotypes were selected to examine the influence of two contrasting temperature regimes during the 24 h incubation (10°C and 15°C) and the subsequent post‐inoculation (17°C and 23°C) periods on latent period and infection efficiency on four susceptible wheat cultivars. In addition, the effect of two contrasting incubation temperatures on urediniospore germination was examined. The results indicated that pathotypes of P. striiformis f. sp. tritici detected after 2002 did not show evidence of adaptation to high temperatures, which suggests that other factors contributed to the observed increased aggressiveness.     

Effects of Simplicillium lanosoniveum on Phakopsora pachyrhizi, the soybean rust pathogen, and its use as a biological control agent

The fungus Simplicillium lanosoniveum was isolated from soybean leaves infected with Phakopsora pachyrhizi, the soybean rust pathogen, in Louisiana and Florida. The fungus did not grow or become established on leaf surfaces until uredinia erupted, but when soybean rust signs and symptoms were evident, S. lanosoniveum colonized leaves within 3 days and sporulated within 4 days. Development of new uredinia was suppressed by about fourfold when S. lanosoniveum colonized uredinia. In the presence of S. lanosoniveum, uredinia became increasingly red-brown, and urediniospores turned brown and germinated at very low rates. Assays using quantitative real time polymerase chain reaction revealed that the fungus colonized leaf surfaces when plants were infected with P. pachyrhizi, either in a latent stage of infection or when symptoms were present. However, when plants were inoculated before infection, there was no increase of DNA of S. lanosoniveum, suggesting that the pathogen must be present in order for the antagonist to become established on soybean leaf surfaces. We documented significantly lower amounts of DNA of P. pachyrhizi and lower disease severity when soybean leaves were colonized with S. lanosoniveum. These studies documented the mycophilic and disease-suppressive nature of S. lanosoniveum.     

Penetration and establishment of Phakopsora pachyrhizi in soybean leaves as observed by transmission electron microscopy

For over 30 years, it has been known that Phakopsora pachyrhizi is unusual in that it penetrates from urediniospores directly through the leaf cuticle without entering stomates. This unusual mode of penetration suggests that disease resistance mechanisms might exist for soybean rust that do not exist for most rust diseases. As a result, we decided to conduct a histological study using transmission electron microscopy to further elucidate the mechanisms of penetration and early establishment of P. pachyrhizi in soybean leaves. Based on our study, it was concluded that P. pachyrhizi utilizes primarily mechanical force, perhaps with the aid of digestive enzymes, to penetrate the cuticle on the leaf surface. However, the lack of deformation lines in micrographs indicated that digestive enzymes, without mechanical force, are used by the penetration hypha to penetrate the outer and inner epidermal cell walls. Digestive enzymes, again indicated by the lack of deformation lines, are used by haustorial mother cells to breach the walls of mesophyll cells to form haustoria. The possibility exists for eventual determination of the precise roles of pressure and digestive enzymes in the development of soybean rust and elucidation of some of the determinants of resistance and susceptibility to this important plant disease.