Localized hemibiotrophy in Colletotrichum: cytological and molecular taxonomic similarities among C. destructivum, C. linicola and C. truncatum

The infection process of hemibiotrophic isolates of Colletotrichum linicola (from flax, Linum usitatissimum ) and C . truncatum (from broad bean, Vicia faba and lentil, Lens culinaris ) was studied by light microscopy. Host surfaces were penetrated directly leading to a symptomless, biotrophic phase characterized by the elaboration of large multilobed, multiseptate, vesicular primary hyphae that were restricted to the initially infected epidermal cells. Biotrophy lasted for the first 48 h of the host-pathogen interaction and was rapidly succeeded by a necrotrophic phase during which narrow, secondary hyphae invaded the surrounding leaf tissues and water-soaked spreading lesions with sporulating, monosetate acervuli were produced on infected host surfaces. Molecular taxonomic analysis of the nucleotide sequences of the amplified D2 and ITS-2 regions of rDNA revealed very close similarities (97–99%) between these isolates and those of C . destructivum obtained from cowpea ( Vigna unguiculata ) and lucerne ( Medicago sativa ), and also of C . truncatum obtained from pea ( Pisum sativum ). This association was consistent with results from a comparative assessment of some in-planta and in-vitro morphological and growth characteristics of these hemibiotrophic fungi. It was concluded that localized hemibiotrophy is an infection strategy utilized predominantly by a closely-related group of pathogens comprising C . destructivum , C . linicola and C. truncatum , and the formation of multilobed primary hyphae restricted to the first penetrated cell might therefore be a key taxonomic character which correlates consistently with ITS sequence data.     

Stomatal penetration of cowpea (Vigna unguiculata) leaves by a Colletotrichum species causing latent anthracnose

The infection process of a Colletotrichum species causing latent infection and anthracnose in cowpea ( Vigna unguiculata ) was studied in seedlings by light and confocal microscopy. Leaf surfaces were extensively colonized by an anastomosing network of germ-tubes and conidia. This epiphytic mycelium produced abundant secondary conidia on short conidiophores. Although melanized appressoria were developed, the host surface was not penetrated directly. The fungus only gained ingress into leaves through stomatal openings, by means of undifferentiated germ-tubes, and slowly colonized the mesophyll by intercellular hyphae, without initially producing visible symptoms. Anthracnose lesions with multisetate acervuli appeared on senescent leaves after a prolonged symptomless period of host colonization lasting > 2 weeks. Analysis of the nucleotide sequences of the amplified D2 and ITS-2 regions of rDNA revealed close similarities (95–96%) between this cowpea pathogen and isolates of C . gloeosporioides from Aeschynomene virginica, Stylosanthes scabra and Mangifera indica. These results, in addition to other morphological and growth attributes, identify this endophytic anthracnose pathogen of cowpea as a Colletotrichum species distinct from C. capsici and C. destructivum .     

Resistance to infection by stealth: <Emphasis Type="Italic">Brassica napus</Emphasis> (winter oilseed rape) and <Emphasis Type="Italic">Pyrenopeziza brassicae</Emphasis> (light leaf spot)

Light leaf spot (Pyrenopeziza brassicae) is an important disease on winter oilseed rape crops (Brassica napus) in northern Europe. In regions where economically damaging epidemics occur, resistance to P. brassicae in commercial cultivars is generally insufficient to control the disease without the use of fungicides. Two major genes for resistance have been identified in seedling experiments, which may operate by decreasing colonisation of B. napus leaf tissues and P. brassicae sporulation. Much of the resistance present in current commercial cultivars is thought to be minor gene-mediated and, in crops, disease escape and tolerance also operate. The subtle strategy of the pathogen means that early colonisation of host tissues is asymptomatic, so a range of techniques and molecular tools is required to investigate mechanisms of resistance. Whilst resistance of new cultivars needs to be assessed in field experiments where they are exposed to populations of P. brassicae under natural conditions, such experiments provide little insight into components of resistance. Genetic components are best assessed in controlled environment experiments with single spore (genetically fixed) P. brassicae isolates. Data for cultivars used in the UK Recommended List trials over several seasons demonstrate how the efficacy of cultivar resistance can be reduced when they are deployed on a widespread scale. There is a need to improve understanding of the components of resistance to P. brassicae to guide the development of breeding and deployment strategies for sustainable management of resistance to P. brassicae in Europe.     

黑胫病弱毒种Leptosphaeria biglobosa通过不同的信号途径诱导油菜抗性

油菜黑胫病是由两种类型病原引起的,分别为A型强致病株(L.maculans)和B型弱致病株,B型现命名为一个新种L.biglobosa。本试验研究了弱致病株诱导的针对强致病株的抗性及其分子信号途径。油菜叶分别用弱致病株子囊孢子预接种和及病害防御反应信号分子水杨酸类似物(acibenzolar-S-methyl, ASM)与维生素K3(menadione sodiumbisulphate,MSB)处理,然后挑战接种强致病株子囊孢子。在控制条件下的试验中,预接种和预处理均推迟了由强致病株引起的病害症状的出现,减小了局部叶和系统叶上的病斑大小。2年的田间试验表明,弱致病株和ASM不仅减少了…     

An iron-regulated ferric reductase associated with the absorption of dietary iron

The ability of intestinal mucosa to absorb dietary ferric iron is attributed to the presence of a brush-border membrane reductase activity that displays adaptive responses to iron status. We have isolated a complementary DNA, Dcytb (for duodenal cytochrome b), which encoded a putative plasma membrane di-heme protein in mouse duodenal mucosa. Dcytb shared between 45 and 50% similarity to the cytochrome b561 family of plasma membrane reductases, was highly expressed in the brush-border membrane of duodenal enterocytes, and induced ferric reductase activity when expressed in Xenopus oocytes and cultured cells. Duodenal expression levels of Dcytb messenger RNA and protein were regulated by changes in physiological modulators of iron absorption. Thus, Dcytb provides an important element in the iron absorption pathway.     

Geographic variation in severity of phoma stem canker and <Emphasis Type="Italic">Leptosphaeria maculans/ L. biglobosa</Emphasis> populations on UK winter oilseed rape (<Emphasis Type="Italic">Brassica napus</Emphasis>)

Phoma stem canker, caused by Leptosphaeria maculans and L. biglobosa, is the most important disease of oilseed rape in Europe. Differences between L. maculans and L. biglobosa in their life-cycles enable the two species to co-exist on oilseed rape crops over a cropping season. This review considers the factors affecting geographic variation in the severity of phoma stem canker epidemics and in the structure of the population of the pathogens in the UK, where the most severe epidemics occur in the south of England and cankers do not develop in Scotland. It is concluded that this variation is directly related to differences in climate, since weather-based models show that stem canker severity increases with increasing winter/spring temperature and temperatures are greater in the south of the UK. It may also be related to differences in pathogen populations, since surveys showed that the proportion of the more damaging L. maculans in stem cankers was greatest in southern England, with most L. biglobosa in northern England. Regional variation in agronomic practices such as cultivar choice and fungicide use may also indirectly influence phoma stem canker severity. Differences in cultivar choice result in differences in L. maculans race structure, which may influence the severity of epidemics. Differences in fungicide use may also influence pathogen populations, since L. maculans and L. biglobosa differ in their sensitivities to different azole fungicides. These factors are discussed in relation to strategies for sustainable production of oilseed rape by adaptation to threats posed by climate change.     

Visual and PCR assessment of light leaf spot (Pyrenopeziza brassicae) on winter oilseed rape (Brassica napus) cultivars

Methods to assess light leaf spot ( Pyrenopeziza brassicae ) on winter oilseed rape cultivars were compared in laboratory, controlled-environment and field experiments. In controlled-environment experiments with seedling leaves inoculated at GS 1,4, the greatest differences in percentage area affected by P. brassicae sporulation were observed with inoculum concentrations of 4 × 10³ or 4 × 10⁴ spores mL⁻¹, rather than 4 × 10² or 4 × 10⁵ spores mL⁻¹, but older leaves had begun to senesce before assessment, particularly where they were severely affected by P. brassicae . In winter oilseed rape field experiments, a severe light leaf spot epidemic developed in 2002/03 (inoculated, September/October rainfall 127·2 mm) but not in 2003/04 (uninoculated, September/October rainfall 40·7 mm). In-plot assessments discriminated between cultivars best in February/March in 2003 and June in 2004, but sometimes failed to detect plots with many infected plants (e.g. March/April 2004). Ranking of cultivar resistance differed between seedling experiments done under controlled-environment conditions and field experiments. The sensitivity of detection of P. brassicae DNA extracted from culture was greater using the PCR primer pair PbITSF/PbITSR than using primers Pb1/Pb2. P. brassicae was detected by PCR (PbITS primers) in leaves from controlled-environment experiments immediately and up to 14 days after inoculation, and in leaves sampled from field experiments 2 months before detection by visual assessment.     

Infection process of Colletotrichum destructivum O'Gara from lucerne (Medicago sativa L.)

The infection process of Colletotrichum destructivum, a cause of anthracnose in lucerne (= alfalfa, Medicago sativa) was studied by light microscopy. At the onset of the host-pathogen interaction, the fungus produced large, multilobed, multiseptate infection vesicles with elongated neck regions. Each infection structure packed the lumen of the initially-infected epidermal cell and remained confined within its walls for 48 h. Subsequently, narrow, invasive secondary hyphae radiated from the multilobed vesicles, grew through the walls of host cells and rapidly colonized the surrounding tissues. Acervuli emerged on the surface of colonized leaves 96 h after inoculation. These observations are discussed in relation to the infection process and specificity of a genetically closely-related isolate of Colletotrichum destructivum causing anthracnose in cowpea (Vigna unguiculata).     

Protection of Brassica seedlings against downy mildew and damping-off by seed treatment with CGA 245704, an activator of systemic acquired resistance

Control of seedling diseases is a major priority in many crop systems. Seed treatments that induce systemic resistance after seedling emergence may be an ideal way to provide protection against disease during the establishment of the crop. CGA 245704, a chemical activator of systemic acquired resistance, was tested as a seed treatment against two Brassica diseases with contrasting infection biologies, the airborne downy mildew pathogen, Peronospora parasitica, and the soilborne fungus, Rhizoctonia solani. Seeds of two Brassica spp. were either imbibed with various concentrations of the compound or imbibed and then dried. Both the imbibition treatment alone and the imbibition treatment followed by seed drying had a significant effect on the sporulation intensity of P. parasitica for all concentrations of the compound used, whereas the imbibition treatment provided some control of damping-off caused by R. solani, with the degree of control being highly dependent on the concentration applied to the seed. Seed treatment with the plant activator CGA 245704 might therefore simultaneously control several seedling diseases, thereby providing a novel option for management of these diseases. © 1998 SCI.