Histology of the infection of tritordeum and its parents by cereal brown rusts

The reactions of tritordeum lines and their Hordeum chilense and Triticum spp. parents to infection by Puccinia recondita f.sp. tritici, P. recondita f.sp. agropyri and P. hordei were studied at the seedling stage. The histological observations indicated that tritordeum behaves as the wheat parent, whatever the H. chilense parental line reaction. Tritordeum is to be considered a host of wheat brown rust where genes for hypersensitive resistance may occur; these are apparently contributed by the wheat parent. Both H. chilense and wheat are highly resistant to barley brown rust, and the reaction of tritordeum is that of the wheat parent with respect to the levels of necrosis associated with the early aborted infection units. The tritordeum reaction is also that of the wheat parent to a rust collected on H. jubatum (putatively P. recondita f.sp. agropyri ), with the susceptibility or resistance of H. chilense being overruled by the wheat parent reaction.     

Histological Characterization of Resistance to Uromyces viciae-fabae in Faba Bean

ABSTRACT Components of resistance to the faba bean rust (Uromyces viciae-fabae) were studied at the histological level in seedlings and adult plants of nine faba bean (Vicia faba) lines differing in their level of resistance. Resistance of these lines was previously shown to be characterized macroscopically by an increased latent period, a decreased colony size, and a relatively decreased infection frequency. In some lines, the resistance also was associated with macroscopically visible necrosis. Histological investigations revealed few differences in spore germination and appressorium formation. Significant levels of aborted stomatal penetration by the rust fungus were found on all resistant lines. However, differences among lines were more evident once the stomata were penetrated by the infection structures. Resistance was mainly due to a restriction of haustorium formation with varying levels of early abortion of the colonies, a reduction in the number of haustoria per colony, and smaller colony size. In addition, necrosis of the host cells associated with infection hyphae was detectable in some lines from the beginning of colony development. This microscopically visible necrosis became stronger from 4 days after inoculation, resulting in a reduced growth of the colony. Differences in resistance levels were more marked in adult plants than in seedlings.     

Specificity of Prehaustorial Resistance to Puccinia hordei and to Two Inappropriate Rust Fungi in Barley

ABSTRACT To elucidate the specificity of prehaustorial resistance to inappropriate rust fungi, we studied two populations of recombinant inbred lines of barley that segregated for partial resistance (PR) to Puccinia hordei and for the resistance to the inappropriate rust species P. recondita f. sp. tritici and P. hordei-murini. PR to P. hordei is prehaustorial and nonhypersensitive, and its level can be assessed accurately by measuring the latent period of the fungus. The resistance to the inappropriate rust species is a combination of prehaustorial (nonhypersensitive) and posthaustorial (hypersensitive) mechanisms. The amount of nonhypersensitive, early abortion of P. recondita f. sp. tritici and P. hordei-murini sporelings reflects the degree of prehaustorial defense to the two inappropriate rust species. All lines showing a long latent period of P. hordei also had a relatively high level of early abortion of the growth of P. recondita f. sp. tritici and P. hordei-murini. This indicates that genes for PR to P. hordei are also effective against these two inappropriate rust species. The reverse was not necessarily true; some lines showing a high level of early abortion of P. recondita f. sp. tritici and P. hordei-murini had a low level of PR to P. hordei. Moreover, lines with a similar level of prehaustorial resistance to P. recondita f. sp. tritici could differ considerably in their prehaustorial resistance to P. hordei-murini. This indicates that genes for prehaustorial resistance may exhibit rust species specificity.     

甘蔗优良新品种(系)抗褐锈病基因Bru1的分子检测及自然抗性评价

甘蔗褐锈病是目前中国蔗区发生最普遍、危害最严重的病害之一,选育和种植抗病品种是防治该病最为经济有效的措施。为明确近年国家甘蔗体系育成的50个新品种(系)对甘蔗褐锈病的抗性,确定其应用潜力,本研究通过在甘蔗褐锈病高发蔗区云南德宏、保山2个区域化试验站,采用田间自然抗性调查与分子标记辅助鉴定抗性基因的方法,于2014年和2015年对中国近年选育的50个优良新品种(系)及2个主栽品种进行抗褐锈病基因Bru1的分子检测及自然抗性评价。田间自然发病调查结果表明,50个优良新品种(系)及2个主栽品种中,高抗至中抗的有34个,占65.38%。其中15个材料表现高抗,占28.85%,16个材料表现抗病,占30.77%,3个材料表现中抗,占5.77%;分子检测结果显示,共29个抗病材料含有抗褐锈病基因Bru1,出现频率为55.77%,表明中国近年选育的优良新品种(系)中抗褐锈病性主要由Bru1控制;其余5个抗病材料均不含抗褐锈病基因Bru1,暗示除了Bru1外,可能还有其他抗褐锈病基因存在。不同系列品种田间感病品种的频率和含抗褐锈病基因Bru1的频率不同,粤糖系列品种感病品种的频率最高达到60%,含Bru1的频率最低,只有30%,抗性最弱;云蔗系列品种感锈病品种的频率最低只有12.5%,含Bru1的频率最高,达到81.25%,抗性最好。本研究结果为深入开展甘蔗抗褐锈病育种,选育和推广优良抗病品种,有效防控甘蔗褐锈病提供了科学依据和优良抗性新品种(系)。     

Inheritance and molecular mapping of barley genes conferring resistance to wheat stripe rust

ABSTRACT Most barley cultivars are resistant to stripe rust of wheat that is caused by Puccinia striiformis f. sp. tritici. The barley cv. Steptoe is susceptible to all identified races of P. striiformis f. sp. hordei (PSH), the barley stripe rust pathogen, but is resistant to most P. striiformis f. sp. tritici races. To determine inheritance of the Steptoe resistance to P. striiformis f. sp. tritici, a cross was made between Steptoe and Russell, a barley cultivar susceptible to some P. striiformis f. sp. tritici races and all tested P. striiformis f. sp. hordei races. Seedlings of parents and F(1), BC(1), F(2), and F(3) progeny from the barley cross were tested with P. striiformis f. sp. tritici races PST-41 and PST-45 under controlled greenhouse conditions. Genetic analyses of infection type data showed that Steptoe had one dominant gene and one recessive gene (provisionally designated as RpstS1 and rpstS2, respectively) for resistance to races PST-41 and PST-45. Genomic DNA was extracted from the parents and 150 F(2) plants that were tested for rust reaction and grown for seed of F(3) lines. The infection type data and polymorphic markers identified using the resistance gene analog polymorphism (RGAP) technique were analyzed with the Mapmaker computer program to map the resistance genes. The dominant resistance gene in Steptoe for resistance to P. striiformis f. sp. tritici races was mapped on barley chromosome 4H using a linked microsatellite marker, HVM68. A linkage group for the dominant gene was constructed with 12 RGAP markers and the microsatellite marker. The results show that resistance in barley to the wheat stripe rust pathogen is qualitatively inherited. These genes might provide useful resistance against wheat stripe rust when introgressed into wheat from barley.     

柔软滨麦草及其杂交后代抗条锈性的组织病理学研究

 对柔软滨麦草及其与感病亲本普通小麦7182的异染色体后代抗小麦条锈菌的组织病理学研究结果显示,柔软滨麦草叶面没有锈菌侵染点形成,且抗病表型为免疫型,因此其抗锈性完全表现为锈菌侵入前的非寄主抗病性。而其后代具有锈菌侵入前、侵入中、侵入后不同阶段的寄主抗锈性,侵入前的抗锈性表现为气孔的运动和气孔下的荧光物对芽管正确定向和附着胞分化的影响,侵入中和侵入后的抗锈性表现为伴随寄主细胞坏死的侵入中断和菌落败育。     

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.     

Real-time PCR-based monitoring of DNA pools in the tri-trophic interaction between Norway spruce, the rust Thekopsora areolata, and an opportunistic ascomycetous Phomopsis sp

The difficulty in subculturing biotrophic fungi complicates etiological studies related to the associated plant diseases. By employing internal transcribed spacer rDNA-targeted quantitative real-time polymerase chain reaction, we now show that the heteroecious rust Thekopsora areolata, commonly associated in natural conditions to sapling shoots and cones of Norway spruce and leaves of wild bird cherry, frequently infects nursery-grown seedlings of the conifer. A spatial sampling scheme was used to investigate seedlings and saplings of Norway spruce showing phloem necrosis: the highest concentration of DNA of T. areolata was recorded in the area with necrotic phloem. The separate analysis of bark and wood tissues suggested that the initial spread of the rust to healthy tissues neighboring the infection site takes place in the bark. A Phomopsis species found to coexist with T. areolata in several seedlings showed very high DNA levels in the upper part of the lesion, and even in the visually healthy proximal tissues above the lesions, which indicates that the ascomycete, most probably a secondary invader following primary infection by T. areolata, has a latent stage during early host colonization. We hypothesize that this hemibiotrophic mode of infection contributes to the successful coexistence of Phomopsis with a biotrophic rust.     

Evaluation of Infection Processes and Resulting Disease Caused by Dendryphion penicillatum and Pleospora papaveracea on Papaver somniferum

ABSTRACT Two pathogenic fungi of opium poppy, Pleospora papaveracea and Dendryphion penicillatum, were isolated from field material in Beltsville, MD. The processes of infection by these two fungi were studied to determine the optimal environmental conditions for infection. Both fungi formed appressoria capable of penetrating directly through the plant epidermal layer. Of the two fungi, P. papaveracea was more aggressive, causing more rapid necrosis. Appressorial formation by P. papaveracea occurred as early as 4 h after application of a conidial suspension to poppy leaves. P. papaveracea formed more appressoria than did D. penicillatum, especially at cool temperatures (7 to 13 degrees C). In greenhouse studies, P. papaveracea caused more damage to opium poppy than did D. penicillatum when applied in 10% unrefined corn oil. In the field, P. papaveracea was more consistent in its effects on opium poppy from a local seed source designated Indian Grocery. P. papaveracea caused higher disease ratings, more stem lesions, and equal or greater yield losses than did D. penicillatum on Indian Grocery. The late-maturing opium poppy variety White Cloud was severely damaged by disease, regardless of formulation or fungal treatment. P. papaveracea was the predominant fungus isolated from poppy seed capsules and the only fungus reisolated from the field the following year. These studies provide a better understanding of the infection process and the differences between these two pathogenic fungi and will be beneficial for the development of the fungi as biological control agents.     

多堆柄锈菌侵染玉米的细胞学及超微结构特征

为明确玉米对多堆柄锈菌Puccinia polysora侵染后病理反应的细胞学特征,利用扫描和透射电镜技术分析了玉米自交系与多堆柄锈菌互作中二者的细胞变化过程。多堆柄锈菌对玉米的侵染主要以直接穿透叶片表皮侵入为主,少量可从气孔和细胞间隙侵入。接种后,病菌夏孢子在感病自交系叶片上快速并大量萌发,在叶表生长蔓延并侵入表皮组织细胞,7 d后形成夏孢子堆;在抗病自交系上,病菌萌发、菌丝生长均受到明显抑制,少量入侵的病菌也由于寄主细胞死亡而导致菌丝和夏孢子干瘪死亡。侵染早期在感病寄主细胞间隙出现菌丝并穿透细胞壁,在胞内产生分枝菌丝,此时寄主细胞结构正常;随着菌丝进一步扩展,叶绿体等结构发生紊乱,被侵染细胞逐渐死亡。在抗病自交系上,接菌24 h后寄主即出现过敏性坏死反应,侵入位点与周围细胞快速坏死,抑制菌丝生长蔓延;叶绿体中清晰可见深色颗粒状物质;72 h后细胞壁外侧产生大量致密的深色结晶体,应为与抗病反应相关的酚类物质。表明抗多堆柄锈菌的玉米材料可能存在2种抗病途径,即寄主与病菌互作中由分子识别引起的免疫反应和病菌侵入后的系统防卫反应。     

紫外线照射对小麦条锈菌生物学效应的研究

 以紫外线(UV)不同剂量照射小麦条锈菌夏孢子,结果表明,其严重削弱夏孢子的存活,剂量效应曲线无明显肩区和尾指数区,小表条锈菌夏孢子对低剂量照射较为敏感;经紫外线照射的夏孢子接种小麦感病品种,其致病性发生改变,潜伏期延长,反应型降低,严重度减少;亦降低了小麦条锈菌的繁殖能力,单叶孢子堆数量减少,孢子堆长度变短,产孢量降低,产孢期缩短。这些生物学效应随照射剂量增加而加强。     

Expression of adult plant resistance and its effect on the development of Puccinia striiformis f.sp. tritici in some Australian wheat cultivars

The onset of adult plant resistance (APR) to Puccinia striiformis was examined in seven Australian wheat cultivars under field conditions and in 14 cultivars under controlled environmental conditions. In most cultivars under field conditions the percentage leaf area affected by stripe rust at mid-tillering (GS 22–26; third leaf) was significantly less ( P < 0.05) than on the more susceptible cultivar Teal. The expression of APR was more conspicuous during tillering to node formation; at these growth stages chlorosis and/or necrosis developed in association with rust colonies on the most resistant cultivars. Under controlled conditions, the primary leaves of all cultivars except Olympic and Flinders displayed some resistance when compared with Teal (lower infection types, longer latent periods and/or less percentage leaf area affected). Studies on the spread of stripe rust from infection foci established in selected cultivars in the field indicated that the resistance detected at early growth stages in Suneca in controlled environmental studies also appeared to be effective in the field.     

Prehaustorial Resistance against Alfalfa Rust (Uromyces striatus) in Medicago truncatula

The legume species Medicago truncatula is gaining interests as a plant for structural and functional genomics that can be used to identify agronomically important genes in crop legumes. Resistance to the alfalfa rust (Uromyces striatus) was studied in a germplasm collection of M. truncatula. Accessions varied in resistance, as expressed by disease severity, but none showed macroscopically visible necrosis. Histological investigations, in selected lines covering the whole range of resistance reactions, revealed little difference in spore germination and none in orientation of germtubes on the leaf surface. However, appressorium formation on the stoma was significantly reduced in some accessions. Differences in resistance among accessions were more evident once the stoma were penetrated by the infection structures. Resistance was mainly due to a restriction of haustorium formation with varying levels of early abortion of the colonies, a reduction in the number of haustoria per colony, and hampered colony growth. In addition, necrosis of the host cells associated with infection hyphae was detectable in some accessions from the beginning of colony development. This information will be useful for eventual mapping and cloning analyses of resistance genes in M. truncatula that will in turn be useful for understanding other legume/rust interactions.     

Expression of resistance to Mycosphaerella pinodes in Pisum sativum

The infection of three resistant and two susceptible inbred lines of Pisum sativum by Mycosphaerella pinodes is described for the first time. Two types of resistance, one expressed in epicotyls and one in leaves, were found in all three resistant lines. On epicotyls of susceptible lines, abundant appressoria and penetrations occurred after a short period of hyphal growth. On epicotyls of resistant lines, hyphae grew extensively but rarely formed appressoria, and these failed to penetrate the cuticle. Attempted penetration was associated with the rapid death of 2–6 epicotyl cells, resembling a hypersensitive reaction. In contrast, resistance of leaves, which was only expressed after penetration, involved localization of the fungus by a mechanism involving delayed leaf cell death. It is suggested that a combination of these two types of resistance might provide effective protection against M. pinodes.     

Tolerance of Senecio vulgaris to Infection and Disease Caused by Native and Alien Rust Fungi

ABSTRACT Plant defense strategies against pathogen attack can be divided into either resistance or tolerance. Variation in tolerance is expressed as differences in the relationship between host fitness (or yield) and the degree of infection. Plant tolerance of pathogen attack remains poorly understood both in terms of its specific mechanisms and in terms of the evolutionary processes by which it has arisen. Theoretical models predict that it is the result of coevolution between host and pathogen, suggesting greater tolerance in interactions with native as opposed to introduced pathogens. Therefore, we quantified and compared the degree of tolerance expressed in the interaction of Senecio vulgaris with the rust fungus Coleosporium tussilginis, which is native to the UK, and the introduced rust fungus Puccinia lagenophorae. We used the reaction norm approach to quantify tolerance and its components. The S. vulgaris-C. tussilaginis interaction expressed a significantly greater degree of tolerance, as reductions in host growth and fitness per unit infection were significantly less than with P. lagenophorae. The key mechanism for this greater tolerance to C. tussilaginis was a significantly smaller reduction in photosynthesis per unit infection than with P. lagenophorae, at both leaf and whole plant scales. There was no significant difference in the relationship between whole plant photosynthesis and host reproduction. We discuss these responses in the context of coevolution for tolerance in host-pathogen interactions.     

Three-way fungal interactions affect the potential biological control of Himalayan balsam,Impatiens glandulifera

Himalayan balsam (Impatiens glandulifera) is one of the most invasive weeds across Europe. The rust fungus, Puccinia komarovii var. glanduliferae has been introduced as a biological control agent, but success has been patchy. Here, we investigated whether mycorrhizal and endophytic fungi can affect rust efficacy and plant growth. Over three experiments, we found that AM fungi and the rust alone or together consistently reduced plant growth, but this depended on the identity of species in the AM inoculum. Meanwhile, AM fungi increased infection frequency of the endophyte Colletotrichum acutatum. Rust inoculation had no detrimental effects on mycorrhizal colonisation or C. acutatum infection, but the latter two fungi reduced rust sporulation. However, plant size was reduced when all three fungal types were present, suggesting that a combined fungal inoculum offers a promising approach for the control of this weed.     

Susceptibility of invasive taxa of European blackberry to rust disease caused by the uredinial stage of Phragmidium violaceum under field conditions in Australia

European blackberry ( Rubus fruticosus agg.) is an aggregate of closely related taxa, with at least 15 taxa naturalized in Australia. Biological control of this Weed of National Significance, using the nonindigenous rust fungus Phragmidium violaceum , is effective when the weather is conducive to multiple cycles of infection, but some blackberry taxa escape severe disease. Thirty-one taxa of naturalized R. fruticosus agg. from southeastern Australia were isolated, their DNA phenotype determined and clones of each taxon inoculated with P. violaceum isolate SA1. Disease development was monitored for at least four generations of uredinia on large potted plants under field conditions. Although variation in mean disease severity appeared continuous over the range of Rubus clones tested, counts of uredinia and telia enabled identification of eight resistant taxa. Fine scale variation in susceptibility to rust disease was observed when different clones of R. leucostachys with the same DNA phenotype were found to express either resistance or susceptibility to P. violaceum (SA1). There were significant differences among 23 Rubus taxa rated as susceptible to rust disease in the mean number of leaves emerging per latent period of uredinia (LELPU). Mean LELPU appeared to account for some of the variation in two measures of mean disease severity observed among susceptible Rubus clones, although the correlation was insignificant (0·10 <  P  > 0·05).     

Effect of the rust fungus Puccinia chondrillina TU 788 on plant size and plant size variability in Chondrilla juncea

Chondrilla juncea is one of the most serious introduced weed species in Australia. Successful biological control has been achieved by the release of the rust fungus Puccinia chondrillina. Although P. chondrillina is thought to reduce the competitive ability of C. juncea by reducing plant size, the precise mechanism by which the pathogen controls its host is poorly understood. In this experiment, we examined the effect of P. chondrillina TU 788 infection on individual plant size, and size variability, in two genotypes (one resistant, one susceptible) of C. juncea. Exposure to P. chondrillina significantly reduced the average size of susceptible C. juncea plants, a reduction apparent in both roots and shoots. Plant size variability of the susceptible C. juncea genotype was also reduced by P. chondrillina. Plant size, and plant size variability of the resistant genotype were unaffected by exposure to rust, or association with rust‐affected plants. The effect of P. chondrillina on plant size in rust‐susceptible C. juncea plants shows how effective bio‐control of this weed may occur in field populations.     

Colletotrichum dematium Produces Phytotoxins in Anthracnose Lesions of Mulberry Leaves

ABSTRACT Colletotrichum dematium, the causal agent of mulberry anthracnose, was examined to produce phytotoxins in vitro and in planta. Raw and autoclaved mulberry leaves infected with the fungus, as well as the fungus incubated with several solid or liquid media, were extracted with acetone. Extracts obtained from the fungus grown on raw and autoclaved mulberry leaves caused brown necrotic lesions on susceptible mulberry leaves when they were placed (10 mul) on the wounded adaxial surface. Whereas, no extracts obtained from media, except inoculated medium containing homogenized mulberry leaves, induced the necrosis, suggesting that the fungus produced phytotoxins in planta and that some components in mulberry leaves may be indispensable substrates for producing the toxins. The phytotoxins obtained from the diseased leaves induced necrosis on nonhost plants leaves as well as on mulberry leaves. The toxins were present in the border of anthracnose lesions on the leaves, and the sensitivity to the toxin correlated with that to the fungus infection in each susceptible or resistant mulberry cultivar. These results suggest that the phytotoxins are host nonspecific and play a role in fungal pathogenesis in the development of the lesions. Four toxic compounds were isolated and purified from anthracnose lesions. However, due to the low yield, the chemical structure of the compounds could not be identified.