Sorghum ergot can develop without local Claviceps africana inoculum from nearby infected plants

Batches of glasshouse-grown flowering sorghum plants were placed in circular plots for 24 h at two field sites in southeast Queensland, Australia on 38 occasions in 2003 and 2004, to trap aerial inoculum of Claviceps africana. Plants were located 20–200 m from the centre of the plots. Batches of sorghum plants with secondary conidia of C. africana on inoculated spikelets were placed at the centre of each plot on some dates as a local point source of inoculum. Plants exposed to field inoculum were returned to a glasshouse, incubated at near-100% relative humidity for 48 h and then at ambient relative humidity for another week before counting infected spikelets to estimate pathogen dispersal. Three times as many spikelets became infected when inoculum was present within 200 m of trap plants, but infected spikelets did not decline with increasing distance from local source within the 200 m. Spikelets also became infected on all 10 dates when plants were exposed without a local source of infected plants, indicating that infection can occur from conidia surviving in the atmosphere. In 2005, when trap plants were placed at 14 locations along a 280 km route, infected spikelets diminished with increasing distance from sorghum paddocks and infection was sporadic for distances over 1 km. Multiple regression analysis showed significant influence of moisture related weather variables on inoculum dispersal. Results suggest that sanitation measures can help reduce ergot severity at the local level, but sustainable management will require better understanding of long-distance dispersal of C. africana inoculum.     

Evaluation of potential biocontrol agents against Claviceps africana in vitro and in vivo

Undiluted culture filtrates of two commercial products of Trichoderma spp., Trichopel and Trichoflow, and two isolates of Penicillium citrinum completely inhibited the conidial germination of macroconidia of Claviceps africana , the cause of ergot or sugary disease of sorghum ( Sorghum bicolor ) in vitro . Similarly, Pseudomonas aeruginosa and Burkholderia cepacia completely inhibited macroconidial germination, with the former being more effective at high dilutions. In contrast, these bacterial isolates failed to inhibit infection in vivo in glasshouse tests with ergot-inoculated sorghum, but all fungal biocontrol agents (including an isolate of Epicoccum nigrum ) reduced the severity of disease (percentage of infected spikelets per panicle), in some cases completely inhibiting the development of ergot. In a second glasshouse trial, optimum control was achieved when the biocontrol agents were applied 3–7 days before inoculation with conidia of C. africana .     

Evaluation of sorghum germplasm used in US breeding programmes for sources of sugary disease resistance*

Ergot or sugary disease of sorghum has become an important constraint in North and South American countries that rely on F₁ hybrid seeds for high productivity. The objective of this research was to determine the vulnerability of various germplasm sources and publicly bred sorghum lines to sugary disease ( Claviceps africana ) in the United States. Flower characteristics associated with sugary disease resistance were also studied. A-/B-line pairs, R-lines, putative sources of resistance and their hybrid combinations with an A₃ cytoplasmic male-sterile source were evaluated using a disease incidence, severity, and dual-ranking system. Trials were planted in a randomized complete block design with three replications and repeated in at least two planting dates. Planting dates and pedigrees had significant effects on overall ranking for resistance. A-lines were most susceptible to sugary disease. R-lines were more susceptible than B-lines with respect to incidence and severity of the disease. Newer releases of A- and B-lines were more susceptible to sugary disease than older releases. Sugary disease reaction of A-lines was a good indicator of disease reaction of B-lines. Tx2737, a popular R-line, was highly susceptible to sugary disease in spite of being a good pollen shedder because the stigma emerged from glumes 2–3 days before anthesis. The combination of flower characteristics associated with resistance were least exposure time of stigma to inoculum before pollination, rapid stigma drying after pollination, and small stigma. An Ethiopian male-fertile germplasm accession, IS 8525, had good levels of resistance. Its A₃ male-sterile hybrid had the highest level of resistance in the male-sterile background. IS 8525 should be exploited in host-plant resistance strategies.     

Susceptibility to ergot in Zimbabwe of sorghums that remained uninfected in their native climates in Ethiopia and Rwanda

Forty-four local Ethiopian and Rwandan sorghums ( Sorghum bicolor ) were observed to remain free of ergot, or had only low incidence, in their natural equatorial latitudes and were potentially of interest, in the design of male-sterile lines for F₁ hybrid breeding, if they possessed a physiologically based resistance mechanism. These sorghums were therefore also investigated under natural and artificial disease pressures in Zimbabwe where unadapted development and inappropriate long daylength prevented flowering in 18 accessions. Of the remaining 16 Ethiopian and 10 Rwandan accessions which flowered, only one from each country remained free of ergot. The susceptibility expressed was ascribed to observed asynchrony of stigma exsertion with anthesis. In the Rwandan accession that persistently remained free of ergot in Zimbabwe, histology of ovules showed pollination before floret gaping, so that a general principle of disease escape due to efficient pollination is proposed for the Ethiopian and Rwandan sorghums in their native climates. The findings emphasize that cleistogamy is a desirable character for avoiding ergot infection in self-fertile sorghums and suggest that the Ethiopian and Rwandan sorghums may not generally be useful for breeding ergot-resistant male-sterile female lines. However, a few accessions deserve more detailed study as a potential genetic resource, before a firm conclusion that all apparent resistance is disease escape owing to efficient pollination.     

Assessment of Diversity in Claviceps africana and Other Claviceps Species by RAM and AFLP Analyses

ABSTRACT Genetic diversity among isolates of Claviceps africana, the sorghum ergot pathogen, and isolates of other Claviceps spp. causing ergot on sorghum or other hosts, was analyzed by random amplified microsatellite (RAM) and amplified fragment length polymorphism (AFLP) analyses. Of the RAM primer sets tested, one revealed polymorphism in C. africana isolates, with Australian and Indian isolates possessing a unique fragment. AFLP analysis, in addition to clearly distinguishing Claviceps spp., revealed polymorphisms in C. africana. A group of isolates from the United States, Puerto Rico, and South Africa exhibited 95 to 100% similarity with one another. Several isolates from Isabela, Puerto Rico were 100% similar to an isolate from Texas, and another isolate from Puerto Rico was identical with one from Nebraska. Australian and Indian isolates showed greater than 90% similarity with isolates from the United States., Puerto Rico, and South Africa. A number of polymorphisms existed in the United States group, indicating that the recently introduced population contains multiple genotypes. Isolates of C. sorghicola, a newly described sorghum pathogen from Japan, were very distinct from other species via RAM and AFLP analyses, as were isolates from outgroups C. purpurea and C. fusiformis. Both RAM and AFLP analysis will be useful in determining future patterns of intercontinental migration of the sorghum ergot pathogen, with the AFLP method showing greater ability to characterize levels of intraspecific variation.     

Windborne spread of ergot disease (Claviceps africana) in sorghum A-lines in Zimbabwe

Ergot disease spread rapidly in Zimbabwe amongst replicated plots of male-sterile sorghum A-lines, from a group of centrally situated and precociously inoculated plants. Prominent secondary conidiation by the pathogen, Claviceps africana , on the surface of exuded honeydew provided airborne spores which were trapped in a Burkard continuous spore trap and showed diurnal peaks of concentration in air close to the primary source of inoculum. The rate of disease spread ( r =0·2; range 0·14–0·58) closely matched that recorded for other plant pathogens such as Phytophthora infestans and Puccinia graminis tritici , and it is concluded that the characteristic secondary conidia of C. africana were the principal epidemiological agents within the experimental area. Ergot spread by windborne secondary conidia has significant epidemiological and economic implications for sorghum hybrid breeding in southern Africa.     

利用抗病品种防治高粱丝黑穗病的研究

1979～1982年鉴定出19个免疫高粱品种,2个免疫恢复系,4个免疫和1个高抗不育系,其中Tx622A免疫不育系的抗病性似为显性。用Tx622A免疫不育系组配的杂交种Tx622A×4003、Tx622A×晋辐1、Tx622A×298/4003、Tx622A×锦恢75为免疫杂交种,Tx622A×208为高抗杂交种,Tx622A×白平为抗病杂交种。1981～1983年用以上抗病高产杂交种进行了20多万亩的防治示范,对丝黑穗病的防治效果均极为显著,由92～100％,平均99.0％,增产粮食1000多万斤。     

Flowering events in relation to smut susceptibility in pearl millet

In field experiments at ICRISAT, selected lines of pearl millet (Pennisetum glaucum) resistant and susceptible to smut (Tolyposporium penicillariae) were evaluated for the timing of flowering events. In smut-resistant lines, the time from the boot-leaf stage (inoculation) to stigma emergence varied in the range 46–120 h, from boot to anther emergence 105–190 h, and from stigma emergence to anther emergence (protogyny period) 59–131 h; in smut-susceptible lines, the corresponding periods were 62–140 h, 146–200 h and 44–120 h, respectively. There were no significant correlations between timing of events and smut severity. Three cytoplasmically male-sterile lines showed longer protogyny periods and higher smut severity than their corresponding maintainer lines. Four lines having short protogyny periods (22–52 h) and resistance to ergot (Claviceps fusiformis) also showed high resistance to smut. Resistance to smut in most ergot-susceptible lines was independent of the timing of flow ering events, but in ergot-resistant lines it could be closely related to flowering events.     

Ergot resistance in sorghum in relation to flowering, inoculation technique and disease development

Ergot is an important disease of sorghum (Sorghum bicolor) in parts of Africa and Asia. Studies were conducted to determine the relationship between flowering biology and ergot infection, and to develop an artificial field-screening technique to identify ergot resistance in sorghum. Spikelets resisted infection after anthesis, but each day's delay in anthesis after inoculation supported 8-3% more ergot. The screening technique consisted of three components: trimming of panicles to remove pollinated spikelets before inoculation, a single inoculation of trimmed panicles, and panicle bagging for 7-10 days. Inoculated panicles were evaluated by a qualitative visual rating method (on a 1-5 scale) and a quantitative spikelet counting method. Selected accessions from the world collection of sorghum germplasm were screened at Karama Research Station, Rwanda, for two seasons and 12 ergot-resistant lines were identified. These were also resistant at ICRISAT Centre, India.     

Genetic diversity of Claviceps africana on sorghum and Hyparrhenia

RAPD markers were used to survey genetic variability among 140 isolates of Claviceps africana collected from Southern Africa, India, Thailand, Australia and the Americas in 1992–2002. Amplified fragment length polymorphisms were determined for a subset of the isolates. Both markers gave similar results in phenetic analysis of genetic distances between haplotypes of different geographical origin. In the Americas, a single RAPD haplotype was found throughout the various countries. The Eastern lineage consisted of two close haplotypes (one from India, the other from Thailand and Australia). Among five specialized isolates of C. africana from the alternative hosts ( Hyparrhenia spp.), three haplotypes were found. Eleven private alleles distinguished the Hyparrhenia population from that on sorghum. rDNA sequences of sorghum and Hyparrhenia isolates differed in three positions. The African sorghum population of C. africana consisted of 10, mostly closely related haplotypes. Low genotypic diversity ( H _E = 0·0337) and the fact that most of the variation originated from between populations ( G _ST = 0·866) suggested founder effects following recent invasion. In Southern Africa, no significant differentiation was found among six populations. Therefore the data were pooled and tested for prevalence of clonal or sexual reproduction. The presence of the over-represented, widespread RAPD haplotype A; gametic disequilibrium (37% loci detected by exact tests); index of association ( I _A) significantly >0; and the high proportion of compatible loci (in the clone-corrected and total data sets found to be 94 and 99%, respectively) support the hypothesis of clonality as the predominant means of reproduction.     

Evaluation of resistance to Phytophthora cinnamomi in seed-grown trees and clonal lines of Eucalyptus marginata inoculated in lateral branches and roots

Seed-grown trees and six clonal lines of 3·5–4·5-year-old Eucalyptus marginata (jarrah) growing in a rehabilitated bauxite mine site in the jarrah forest were underbark-inoculated on lateral branches (1995) or simultaneously on lateral branches and lateral roots (1996) with isolates of Phytophthora cinnamomi in late autumn. Individual seedlings from which the clonal lines were derived had previously been assessed as either resistant (RR) or susceptible (SS) to P. cinnamomi . At harvest, the acropetal lesion and colonization lengths were measured. Overall, the length of colonization in roots and branches was more consistent as a measure of resistance than lesion length, because colonization length recorded the recovery of P. cinnamomi from macroscopically symptomless tissue ahead of the lesion which, on some occasions, was up to 6 cm. In both trials, one RR clonal line was able to contain the P. cinnamomi isolates consistently, as determined by small lesion and colonization lengths in branches and roots. In contrast, the remaining two RR clonal lines used in both trials were no different from the SS line in their ability to contain lesions or colonization. These latter two RR lines may therefore not be suitable for use in rehabilitation of P. cinnamomi -infested areas. Differences in lesion and colonization lengths among P. cinnamomi isolates occurred only in the 1995 trial. Colonization and lesion lengths in branches were up to eight times greater in 1996 than in 1995, but the relative rankings of clonal lines were consistent between trials. Although colonization was always greater in branches than roots, the relative rankings of the lines were similar between branch and root inoculations. Branch inoculations are a valid option for testing the resistance and susceptibility of young jarrah trees to P. cinnamomi .     

Patterns of release of the secondary conidia of Claviceps africana, the sorghum ergot pathogen in Australia

Trials were conducted in southern Queensland, Australia between March and May 2003, 2004 and 2005 to study patterns of hourly and daily release of the secondary conidia of Claviceps africana and their relationships with weather parameters. Conidia were trapped for at least one hour on most (> 90%) days in 2003 and 2004, but only on 55% of days in 2005. Both the highest daily concentration of conidia, and the highest number of hours per day when conidia were trapped, were recorded 1–3 days after rainfall events. Although the pattern of conidial release was different every day, the highest hourly conidial concentrations occurred between 10·00 hours and 17·00 hours on 73% of all days in the three trials. Hours when conidia were trapped were characterized by higher median values of temperature, windspeed and vapour pressure deficit, lower relative humidity, and leaf wetness values of 0%, than hours when no conidia were recorded. The results indicate that fungicides need to be applied to the highly ergot-susceptible male sterile (A-) lines of sorghum in hybrid seed production blocks and breeders' nurseries as soon as possible after rainfall events to minimize ergot severity.     

Novel infection strategies of Colletotrichum acutatum on ripe blueberry fruit

The infection and colonization process of Colletotrichum acutatum on ripe blueberry fruit from two cultivars with different susceptibility to anthracnose were examined using light and confocal laser scanning microscopy. Ripe fruit from susceptible cv. Jersey and resistant cv. Elliott were drop-inoculated with a conidial suspension of C. acutatum, and epidermal peels were evaluated at selected times after inoculation and incubation. Results from pre-penetration studies demonstrated that there were significant differences in the rate of formation of melanized appressoria between the two cultivars, with the rate of formation being faster in the susceptible one. In both cultivars, penetration by the pathogen occurred via appressoria 48 h post-inoculation (hpi). However, in the susceptible cv. Jersey, C. acutatum then adopted an intracellular hemibiotrophic-like infection strategy, whereas in the resistant cv. Elliott subcuticular intramural-like infection occurred. In cv. Jersey by 108 hpi, intracellular growth of the pathogen led to the formation of numerous acervuli, with orange conidial masses. By 120 hpi, the conidial masses had coalesced covering the entire inoculated area. In cv. Elliott, acervuli were not seen until 144 hpi and contained few conidia. These results demonstrate for the first time the ability of C. acutatum to adopt a different infection and colonization strategy depending on the susceptibility of the host tissue being colonized.     

酿酒高粱品种、组合及亲本的丝黑穗病抗性鉴定

为了明确不同类型的酿酒高粱材料对西南地区丝黑穗病的抗性和促进抗丝黑穗病品种的选育,采用土壤接菌法对107份材料进行了丝黑穗病抗性鉴定;根据抗性鉴定结果,比较了酿酒高粱杂交组合与其亲本对丝黑穗病的抗性,并对供试菌种进行了致病力分析.结果显示:在感病对照恢1植株平均病株率为67.65％时,有13个不育系、6个保持系、10个恢复系、28个杂交组合的病株率为0,达免疫水平,其中,酿酒高粱不育系和杂交组合所占比例较大,分别为86.67％和75.68％;恢复系中免疫类型所占比例较小,为24.39％;常规品种和组合中无免疫类型;杂交高粱对丝黑穗病的抗性属显性遗传;泸州丝黑穗病菌具有较强的致病力.     

山西高粱品种(系)的花后抗旱性评价

以22份山西历年审定(鉴定、登记)的高粱品种及预登记的高粱品种为参试材料,通过自然干旱(DS)和正常灌溉(NI)两种处理方式,采用大田自然鉴定法,调查农艺性状、品质性状和光合性状3类主要性状指标。结合隶属函数(U)和干旱敏感指数(S)的聚类分析法及热图法对材料进行分类,对抗旱指标进行综合评价。结果表明：在干旱胁迫下,高粱品种除了穗柄伸出长平均增长5.26 cm和单宁含量平均增加0.09%,其余性状指标均出现下降;除净光合速率外,其余性状指标之间均存在一定程度的相关性。利用以上两种方法,筛选出极抗品种3份(S9、S10和S7即晋杂108、晋杂22号和晋杂101),极弱品种1份(S17即晋杂4号)。根据干旱敏感指数计算,主成分分析为9个性状,主成分累计贡献率84.662%,产量和单穗粒重是决定第一主成分的主要性状,贡献率为17.999%。干旱胁迫对高粱品种(系)花后主要性状指标均有显著影响。不同高粱材料间抗旱性差异显著,应用两种抗旱性评价方法,可以较为准确地评价参试高粱品种(系)的抗旱性。产量、单穗粒重、株高和倒二叶叶宽可以作为评价高粱花后抗旱性的主要形态指标。     

Screening of Lycopersicon spp. for glyphosate and/or Orobanche aegyptiaca Pers. resistance

A major screening programme was conducted to determine which Lycopersicon spp. (if any) have practical levels of resistance to glyphosate; the same lines were also tested for resistance to Orobanche aegyptiaca. A majority of the lines investigated showed high susceptibility to glyphosate applied at 37?5 g a.i. ha^?1 in a spray volume of 250 1 ha^?1. Only 41 of 1457 lines screened showed fresh weights of treated plants not significantly different from those of untreated plants at a probability level of 80%. Repeated screening of some selected Lycopersicon lines in the greenhouse and the field, however, indicated that only a few had inherent partial resistance to glyphosate. Giyphosate was less toxic to the test plants with increases in spray volume. Among 1361 Lycopersicon lines tested for resistance to O. aegyptiaca, only minor differences in susceptibility were observed. All lines were susceptible to varying degrees. The chances, therefore, of finding complete resistance in tomato to either glyphosate or Orobanche spp. alone are small. However, combining partial resistance to glyphosate and O. aegyptiaca in a breeding programme to produce new varieties might still prove to be economically useful even in fields heavily infested with the parasite.     

Seedling and adult plant resistance to downy mildew (Peronospora parasitica) in cauliflower (Brassica oleracea convar. botrytis var. botrytis)

Seedlings of six cauliflower cultivars ( Brassica oleracea convar. botrytis var. botrytis ) were assessed for resistance to a Danish isolate of Peronospora parasitica , under controlled conditions. Resistance, characterized by restricted sporulation and necrotic dark flecks at the inoculation site on the cotyledons, was expressed in the hybrids 9306 F1, 9311 F1, and the open pollinated cultivar Perfection. Testing of the parent lines and F2 generations of the two resistant hybrids suggested that resistance was a dominantly inherited trait controlled by a single gene. Inoculation of the cultivars with seven isolates, from different geographical origins, showed that the resistance was isolate specific. The two hybrid cultivars expressing cotyledon resistance and two hybrids expressing susceptibility were assessed for adult plant resistance under field conditions. The AUDPC (Area Under the Disease Progress Curve), based on disease incidence and severity, revealed significant differences between the cultivars. At harvest, the cultivars exhibited significantly different levels of defoliation and curd attack. The cultivars 9306 F1 and 9311 F1 showed high levels of resistance in all assessments, whereas the two cultivars exhibiting susceptibility at the seedling stage, 9304 F1 and 9305 F1, also exhibited susceptibility through the adult plant stage. Thus, the resistance exhibited under field conditions resembled that identified at the seedling stage under controlled conditions. The results suggest that cotyledon resistance similar to that described could provide resistance throughout the adult plant stage, including curds.     

Evaluation of the susceptibility of modern,wild, ancestral,and mutational wheat lines to Septoria tritici blotch disease

Globally, bread wheat production is threatened by fungal diseases, including the devastating disease Septoria tritici blotch (STB). Given the global importance of STB, and the difficulty in identifying novel sources of resistance to this disease, we screened a variety of wheat genotypes, including wild, ancestral, and mutagenized lines, for their STB response. This delineated a panel of wild wheat relatives and Watkins collection lines with exceptional resistance to a range of Zymoseptoria tritici isolates, some of which are highly virulent on modern, elite wheat varieties. Additionally, we characterized the STB susceptibility of 500 lines of the wheat cultivar Cadenza TILLING population and developed backcross derivatives of two TILLING lines that show dominant partial resistance to STB. These backcross lines are partially resistant to multiple isolates of Z. tritici, and, with the wild and ancestral lines identified, provide a useful reservoir of STB-resistant germplasm for use in wheat breeding programmes.     

Infection of wheat spikes by <Emphasis Type="Italic">Fusarium avenaceum</Emphasis> and alterations of cell wall components in the infected tissue

The infection process of Fusarium avenaceum on wheat spikes and the alteration of cell wall components in the infected host tissue were examined by means of electron microscopy and cytochemical labelling techniques following spray inoculation at growth stage (GS) 65 (mid-flowering). Macroconidia of the pathogen germinated with one to several germ-tubes 6–12 h after inoculation (hai) on host surfaces. The germ-tubes did not penetrate host tissues immediately, but extended and branched on the host surfaces. Hyphal growth on abaxial surfaces of the glume, lemma and palea was scanty 3–4 days after inoculation (dai) and no direct penetration of the outer surfaces of the spikelet was observed. Dense mycelial networks formed on the inner surfaces of the glume, lemma, palea and ovary 36–48 hai. Penetration of the host tissue occurred 36 hai by infection hyphae only on the adaxial surfaces of the glume, lemma, palea and upper part of ovary. The fungus penetrated the cuticle and hyphae extended subcuticularly or between the epidermal wall layers. The subcuticular growth phase was followed by penetration of the epidermal wall, and hyphae spread rapidly inter- and intracellularly in the glume, lemma, palea and ovary. During this necrotrophic colonization phase of the wheat spike, a series of alterations occurred in the host tissues, such as degeneration of cytoplasm and cell organelles, collapse of host cells and disintegration of host cell walls. Immunogold labelling techniques showed that cell walls of spike tissues contained reduced amounts of cellulose, xylan and pectin near intercellular hyphae or infection pegs compared to walls of healthy host tissues. These studies suggest that cell wall degrading enzymes produced by F. avenaceum facilitated rapid colonization of wheat spikes. The different penetration properties of abaxial and adaxial surfaces of the spikelet tissues as well as the two distinct colonization strategies of host tissues by F. avenaceum are discussed. The penetration and colonization behaviour of F. avenaceum in wheat spikelets resembled that of F. culmorum and F. graminearum, although mycotoxins produced by F. avenaceum differed from those of the latter two Fusarium species.