Effects of Leaf Age, Inoculum Dose and Freezing on Development of Chocolate Spot (Botrytis fabae) Lesions on Field Bean (Vicia faba) Leaves

Botrytis fabae spore suspensions containing c. 1, 10, 10², 10³, 10⁴, 10⁵, or 10⁶ spores/ml were used to inoculate 5, 17 or 30-day-old field bean leaves. The percentages of the leaf areas covered by, chocolate spot lesions and the percentages of the leaf areas bearing conidiophores were assessed 1, 6, 12, 14, and 19 days after inoculation. The percentage of the area covered by lesions and the percentage of the area bearing conidiophores (logit-transformed) increased linearly with increasing spore concentration (log₁₀-transformed). The proportions of leaf areas covered by lesions and bearing conidiophores were both greater on 17 and 30-day-old leaves than on 5-day-old leaves. The rate of lesion growth increased with both increasing inoculum dose and increasing leaf age. Generally there was no interaction between the effects of leaf age and the effects of inoculum dose on either lesion growth or sporulation. Two days after inoculation with suspensions of either 10⁴ or 10⁶ spores/ml, 7-day-old leaves grown at 15°C were transferred to –16°C or 2.5°C or kept at 15°C for 4 days. Two days later more spores had been produced on leaves which had been frozen (–16°C) than on, leaves kept at 2.5°C.     

Influence of temperature and duration of leaf wetness on infection of Acyrthosiphon kondoi with Erynia neoaphidis

Bioassays were carried out to examine the influence of temperature and duration of leaf wetness on the infectivity of an isolate of Erynia neoaphidis for its aphid host Acyrthosiphon kondoi. Preliminary experiments demonstrated that primary spores produced in vitro were as infectious as those formed in vivo. No consistent effect of temperature on infectivity of primary spores could be detected. The time taken to kill an aphid increased as temperature decreased, from 3–5 days at 20 °C to 12–15 days at 8 °C, suggesting a threshold for disease development of 4 °C. Increasing duration of the period of leaf wetness up to 24 h after inoculation increased the final level of infection. At 20 °C, a minimum moisture period of 3 h was required for infection with maximum infection occurring after about 7 h. These times increased slightly at 15 °C but extending to 7 and 16 h respectively at 10 °C. The epizootiological implications of these results are discussed with reference to previously published data on in vivo production of primary spores of E. neoaphidis.     

A Comparison of the Disease Reactions of Stems and Detached Leaves of Soil and in vitro Grown Plants and Regenerants of Oilseed Rape to Leptosphaeria maculans and Protocols for Selection for Novel Disease Resistance

Protocols for selecting plant tissues of winter oilseed rape (Brassica napus subsp. oleifera) with resistance to Leptosphaeria maculans by either stem or leaf inoculation of both soil and in vitro grown plant material are described. The stem inoculation procedure gave good correlation (r = 0. 92) between the 50 day stem disease scores of eight out of nine cultivars of soil grown winter oilseed rape inoculated with isolate 41A4 of L. maculans and the N. A. B. esistance ratings or resistance data from field trials. The exception was the cultivar Liradonna. Inoculation of stems of five cultivars with isolates 41A4, 433 and 478 indicated a range of isolate virulence 478 > 41A4 > 433. This was the inverse of that observed in leaf inoculations. Application of the stem inoculation procedure to in vitro shoot cultures allowed differentiation of resistant and susceptible cultivars, including the cultivar Liradonna, after 20 days incubation at 20°C. The protocol was also applicable to plantlets regenerated from thin cell layer explants grown in vitro. Inoculations with isolate 433 allowed the differentiation of resistant, intermediately resistant and susceptible leaf material of soil grown plants, when leaf discs from young leaves were incubated on water agar supplemented with BAP (1 × 10^?5 M) at 25°C for 10 days. Intermediately resistant leaves were resistant after 10 days and susceptible after 15 days of incubation. Leaves of shoot cultures grown in vitro were more susceptible than the corresponding soil grown material. However, inoculation of old leaves with isolate 41A4 (an isolate of less virulence on leaves than 433) distinguished the cultivars after 15 days of incubation. These protocols allow the accurate assessment of resistance to L. maculans at the stem or leaf level and are of use in traditional as well as in vitro selection programmes.     

Effects of water potential and temperature on the development of eyespot lesions in wheat

Growth of Pseudocercosporella herpotrichoides on potato dextrose agar at water potentials from -0.5 to -6.9 MPa was optimal at 20°C. At 12 and 20°C, six isolates of P. herpotrichoides grew more rapidly at -0.5 to -2 MPa than at -6.9 MPa. Wheat plants inoculated with P. herpotrichoides and grown in columns of soil at either 15 or 20°C developed more severe eyespot lesions under a heavy watering regime than under medium or light watering regimes. P. herpotrichoides penetrated leaf sheaths of inoculated plants grown in compost more rapidly at 10°C night/15°C day temperatures than at 5/10°C; death of leaf sheaths was also more rapid and consequently there were fewer living infected leaf sheaths at 10/15°C than at 5/10°C. Irrigating for 5 wk before harvest increased the severity of eyespot lesions in a 1983 wheat crop.     

Effects of Temperature and Leaf Weness on the Latent Period of Rbynchosporium Secalis (Leaf Blotch) on Leaves of Winter Barley

Effects of temperature and leaf wetness on the latent period of Rhynchosporium secaits (leaf blotch) on winter barley were examined in controlled environment experiments. At 100% relative humidity (continuous leaf wetness) the mean length of the latent period was c.24 days at 5°C, c. 19 days at 10°C, c. 16 days at l5°C and c. 13 days at 20°C. The mean number of days between the appearance of the first and the last lesions was c. 13 days at 5°C, c. 6 days at 10°C, c. 5 days at 15°C and c. 3 days at 20°C. A negative curvilinear regression of latent period on temperature accounted for 99% of the variance. The mean area of lesions per leaf was 38 mm² at 5°C, 46 mm² at 10°C, 24 mm² at 15°C and 24 mm² at 20°C. At 10°C, after a 48 h wet infection period, the interruption of leaf wetness for 5 or more days at any time during the next 15 days of the latent period did not decrease subsequent lesion area. However, absence of leaf wetness after these 15 days, at the onset of sporuiation, did decrease the area of lesions which developed.     

Recombinant chromosomes of advanced backcross plants between Allium cepa L. and A. fistulosum L. revealed by in situ hybridization

Cytological analysis of (Allium cepa L.×Allium fistulosum L.)×A. cepa L. F₁BC₃ plants revealed most plants were diploid with 16 chromosomes. Karyotypes of these plants showed recombinant chromosomes. Fluorescence and genomic in situ hybridization patterns of interspecific F₁ hybrid and F₁BC₃ plants revealed A. fistulosum chromosomes or chromosomal segments. A highly repetitive 376-bp DNA sequence and genomic DNA of A. fistulosum revealed similar telomeric hybridization sites when hybridized onto A. fistulosum chromosomes. Cytogenetic evidence showed that A. fistulosum DNA has recombined into the A. cepa genome. Received: 20 October 1999 / Accepted: 11 November 1999     

Effects of environmental factors on growth of lesions on field bean leaves infected by Botrytis fabae

Chocolate spot lesions increased in size only slowly when the relative humidity of the air was below 66%. Following a lag phase immediately after infection the rate of increase was linear and proportional to humidity between c. 70% and 100% r.h. Lesions on leaflets kept at 70% r.h. for 8 h and at 100% r.h. for 16 h/day increased in size at only 27% of the rate of those at continuous 100% r.h. The optimum temperature for lesion growth was between 15 and 22 °C, the minimum <4 °C and the maximum c. 30 °C. Humidity did not interact with temperature between 10 and 20 °C. Neither light intensity nor a film of water over the leaves affected lesion growth. These findings are discussed in relation to meteorological data and field observations. The possible mechanisms whereby humidity affects lesion growth did not appear to be related to CO₂ and O₂ concentrations nor to the overall water potential of the leaf. Preliminary evidence is presented for the production of phytotoxins within lesions.     

Production of interspecific hybrids between Allium fistulosum L. and A. macrostemon Bunge through ovary culture

Interspecific hybridization is an effective method to generate a new crop in a short time that exhibits available traits. Nine interspecific hybrids between Allium fistulosum and A. macrostemon were produced through reciprocal crossings by ovary culture. This is the first report of hybrid development in the cross-combination. When A. macrostemon, a vegetative propagated plant, was used as the mother plant, no hybrid was obtained, but it was obtained by removal of bulbils and separation of the flower stalk, as with cut flowers. Ovary culture was performed on phytohormone-free B5 medium modified by Dunstan and Short (BDS) medium containing 30 g l⁻¹ sucrose 7 days after pollination. Germinated embryos were transferred to BDS medium containing 1 mg l⁻¹ 6-benzylaminopurine and 15 g l⁻¹ sucrose for 21 days. The plantlets were subcultured on phytohormone-free B5 medium for subsequent rooting. Hybridity was confirmed using cleaved amplified polymorphic sequence analysis of the rDNA internal transcribed spacer region and chromosome observation. In addition, the hybrids showed intermediate traits: they had fistulous leaves like those of A. fistulosum and bulbs like those of A. macrostemon. They displayed vigorous growth and propagated vegetatively by tillering. These results show the important discovery that vegetatively propagated plants are useful for cross breeding.     

Epidemiology of Cladosporium allii and Cladosporium allii-cepae, leaf blotch pathogens of leek and onion.

Conidia of Cladosporium allii and C. allii-cepae germinated over the temperature range 2–30°C on agar with optimal responses at 15–20°C (C. allii) and 20°C (C. allii-cepae). Conidia of both fungi germinated in water and at c. 100% relative humidity (r.h.) but not at lower humidities on leaf and glass slide surfaces. Germination was more rapid when spores were applied dry to agar or leaves than when applied in water or nutrient solution. More lesions developed when conidia of C. allii-cepae were deposited dry on onion leaf discs or leaf surfaces than when they were applied suspended in water. Conidia of both fungi required 18–20 h at c. 100% r.h. to germinate and infect when applied dry to leaves. Damaging the leaves or the addition of nutrients to the leaf surface increased the incidence of infection by C. allii-cepae compared to controls. Inoculated onion bait plants placed out-of-doors developed infection after at least 17 h at c. 100% r.h. or with leaf wetness. Similar conditions were necessary for infection of bait plants exposed in onion and leek crops infected by C. allii-cepae and C. allii respectively. Disease development and spread of infection occurred at different rates over the same period in two different cultivars of leeks, with spore concentrations increasing in proportion to disease. Spore numbers in the air fell considerably when infected leeks were ploughed under.     

In vitro spore germination and infection of cultivars of Rubus and Rosa by downy mildews from both hosts

The cardinal temperatures for in vitro germination of conidia of imported and indigenous isolates of downy mildew from hosts in the genera Rubus and Rosa were similar. A high percentage of conidia germinated above 2°C and germination remained between 80% and 90% up to 15°C or 20°C, depending on the isolate. The highest incidence of disease on leaf disks of Tummelberry (blackberry × red raspberry) inoculated with an isolate of Peronospora rubi occurred at c. 15°C, with infection over a range from 2°C to 28°C. Tests on leaf disks in vitro, and leaflets of primocane and lateral shoots in plastic tunnels, with three hybrid berry (blackberry x red raspberry), six blackberry and nine red raspberry cultivars showed the hybrid berries to be most susceptible. In a plastic tunnel infected drupelets of red raspberry fruits developed more slowly and failed to ripen evenly compared with uninfected drupelets. Similar malformation of infected fruits occurred in a plantation of Tummelberry. An isolate of P. rubi attacked severely both Tummelberry and rose cv. Can Can. Fluorescence microscopy after staining with aniline blue showed that leaf disks of Tummelberry were extensively colonised by intercellular mycelium of P. sparsa isolated from rose, even though sporulation was sparse or absent. This supports the view that P. rubi and P. sparsa may be conspecific. Oospores of P. rubi were found routinely within leaf disks of Rubus cultivars inoculated in vitro and once in naturally infected leaflets of Tummelberry.     

Chromosome characteristics and behavior differences in Allium fistulosum L., A. cepa L,their F1 hybrid,and selected backcross progeny

Mitotic and meiotic studies were performed on Allium fistulosum, A. cepa, their F₁ hybrid, and ten selected backcross (BC)₁ plants [(A. fistulosum x A. cepa) x (A. cepa)]. Each BC₁ plant had at least one A. cepa isozyme allele (Pgi, Idh, or Adh). Chromosome morphology and behavior differed among plants. Meiocytes were observed with one, two, or three bridges and/ or fragments, indicating at least three paracentric inversions between A. fistulosum and A. cepa. Unusual crossing over and multivalent associations suggest that the 5F subtelocentric chromosome of A. fistulosum is involved in at least one translocation. The number of bridges and fragments and multivalent associations varied between the F₁ hybrid and BC₁ progenies. The F₁ hybrid and all BC₁ progenies were either sterile or had very little seed set. Fertility was not restored in any of the selected BC₁ plants.The use of trade names does not imply endorsement of the products named nor criticism of similar ones not named. This research was supported by the New Mexico Agricultural Experiment Station     

Genetic analysis of the interaction between <Emphasis Type="Italic">Allium</Emphasis> species and arbuscular mycorrhizal fungi

The response of Allium cepa, A. roylei, A. fistulosum, and the hybrid A. fistulosum × A. roylei to the arbuscular mycorrhizal fungus (AMF) Glomus intraradices was studied. The genetic basis for response to AMF was analyzed in a tri-hybrid A. cepa × (A. roylei × A. fistulosum) population. Plant response to mycorrhizal symbiosis was expressed as relative mycorrhizal responsiveness (R′) and absolute responsiveness (R). In addition, the average performance (AP) of genotypes under mycorrhizal and non-mycorrhizal conditions was determined. Experiments were executed in 2 years, and comprised clonally propagated plants of each genotype grown in sterile soil, inoculated with G. intraradices or non-inoculated. Results were significantly correlated between both years. Biomass of non-mycorrhizal and mycorrhizal plants was significantly positively correlated. R′ was negatively correlated with biomass of non-mycorrhizal plants and hence unsuitable as a breeding criterion. R and AP were positively correlated with biomass of mycorrhizal and non-mycorrhizal plants. QTLs contributing to mycorrhizal response were located on a linkage map of the A. roylei × A. fistulosum parental genotype. Two QTLs from A. roylei were detected on chromosomes 2 and 3 for R, AP, and biomass of mycorrhizal plants. A QTL from A. fistulosum was detected on linkage group 9 for AP (but not R), biomass of mycorrhizal and non-mycorrhizal plants, and the number of stem-borne roots. Co-segregating QTLs for plant biomass, R and AP indicate that selection for plant biomass also selects for enhanced R and AP. Moreover, our findings suggest that modern onion breeding did not select against the response to AMF, as was suggested before for other cultivated species. Positive correlation between high number of roots, biomass and large response to AMF in close relatives of onion opens prospects to combine these traits for the development of more robust onion cultivars.     

Apical Growth Cessation and Shoot Tip Abscission in Salix

Time course of apical shoot growth and shoot tip abortion in northern ecotypes (lat. 69°39′N, long. 18°37′E) of Salix pentandra and S. caprea have been investigated. In trees more than 15 years old growing under natural climatic conditions apical growth cessation and shoot tip abortion normally occurred in June-July when the day length still was 24 h. Application of GA₃, in spring to the apex effectively delayed growth cessation and shoot tip abortion. Application of kinetin was without effect. First-year seedlings of both species grew continuously at temperatue of 9 to 24°C in 24 h photoperiod. Short days induced apical growth cessation, but two to four (S. pentandra) or three to five (S. caprea) weeks of 12 h photoperiod were required to stop the elongation growth. The results indicated that the critical photoperiod for apical growth cessation in the used ecotype of S. pentandra was 16 to 18 h at 18°C. Short days had a minor effect only on the formation of apical leaf primordia in small seedlings. Development of axillary buds and radial growth were stimulated by short days when compared with long days. Small seedlings of both species (3 to 8 cm high at the start) formed terminal buds in short days, but in large seedlings (more than about 15 cm high) apical growth cessation was accompanied by shoot tip abortion. Abscisic acid applied to the apex or through a leaf did not induce growth cessation in S. pentandra seedlings grown in continuous light. The growth retardants CCC, B-9 and Phosphon D reduced growth rate under continuous light and induced shoot tip abortion in some plants. The effect of CCC was counteracted by GA₃. Apical growth cessation in short days was significantly delayed by a single GA₁ application.     

Resistance toMelampsora larici-epitea leaf rust inSalix: analyses of quantitative trait loci

Quantitative resistance ofSalix toMelampsora larici-epitea leaf rust was studied in 2Salix mapping populations. One population was a backcross between aS. schwerinii ×S. viminalis hybrid andS. viminalis, and the other was an F₂ population betweenS. viminalis andS. dasyclados. A leaf disc bioassay was used to study the components of quantitative resistance (latent period, uredinia number, and uredinia size) to 3 isolates of the leaf rust. The analysis of quantitative trait loci (QTLs) revealed 9 genomic regions in the backcross population and 7 genomic regions in the F₂ population that were important for rust resistance, with QTLs explaining 8–26% of the phenotypic variation. An important genomic region was identified for the backcross population in linkage group 2, where QTLs were identified for all resistance components for 2 of the rust isolates. Four of the QTLs had overlapping mapping intervals, demonstrating a common genetic background for latent period, uredinia diameter, and uredinia number. QTLs specific to some rust isolates and to some resistance components were also found, indicating a combination of common and specific mechanisms involved in the various resistance components. Breeding implications in relation to these findings are discussed.     

Effect of Temperature and Air Humidity on Multiplication of Xanthomonas campestris pv. undulosa and Symptom Expression in Susceptible and Field-Tolerant Wheat Genotypes

Multiplication of Xanthomonas campestris pv. undulosa in the leaf tissues of susceptible and field-tolerant wheat genotypes was determined at the secdling stage, under different incubation conditions of temperature and air humidity. Dry air conditions did not hamper the multiplication of the population trend. Symptoms were not observed at 15°C. The occurrence of lesions coincided with populations of baeteria that had reached an estimated threshold of 10⁸ colony forming units/leaf. Differences in populations between genotypes Alondra and Pavon were only noticeable at 25°C and dry air conditions, without correlation to differences in symptom expression. Considering these two genotypes, resistance to bacterial leaf streak did not seem to operate at the seedling stage when plants were inoeulated by infiltration. In both genotypes, lesions did not extend significantly. An aetive mechanism stopping bacterial growth which is apparently temperature-dependent or a saturation of the, target sites was possibly involved, as shown in the different experiments by the fit-curves calculated for Alondra and Pavon. This study provided information on the role of climatic factors in relation to epidemics. Temperature appeared to have a major effect on the disease and results confirmed that the risk of bacterial leaf streak prevails in warmer non-traditional wheat growing areas. Several types of resistance are likely to exist in the X. e. pv. undulosa/wheat system.     

A cotyledon double inoculation technique for evaluating resistance to anthracnose (Colletotrichum orbiculare) and scab (Cladosporium cucumerinum) in cucumber

A technique for simultaneous inoculation of cucumber cotyledons with Colletotrichum orbiculare race 1 and Cladosporium cucumerinum has been developed. The procedure permitted both resistant and susceptible plants to be recovered. Seedlings were grown at 20°C and inoculated 24 h after emergence with Colletotrichum orbiculare (200 spores in 2 μ1 of water) and Cladosporium cucumerinum (1000 spores in 5 μ1 of water) followed by 48 h of incubation in the dark at 20°C and 100% r.h., and 48 h in a 20°C lighted growth chamber. Seedlings were then moved to a growth chamber at 21°C at night and at 26°C during the day for 4 days and plants were rated as resistant or susceptible 8 days after inoculation. No interference in the expression of resistance or susceptibility of cultivars to either pathogen was detected in simultaneous inoculations.     

A glasshouse screening method to assess resistance in onions to damage by onion fly (Delia antiqua) larvae

Factors influencing the resistance of onion (Allium cepa) cultivars to onion fly (Delia antiqua) damage were investigated. These included different environments, methods of plant raising and egg inoculation, and plant size. A glasshouse compartment maintained at 14 to 37 °C was as satisfactory for assessing the resistance as a controlled environment. The basic test unit comprised 10 onion seedlings raised in a 400 × 50 × 50 mm deep plastic trough and inoculated with 50 onion fly eggs from a laboratory culture. Resistance was assessed by counting wilted plants every two or three days for 21 days after egg inoculation. The reciprocal of number of days for each plant to wilt (wilting rate) was an appropriate variate for statistical analysis, adjusted for the length of the longest leaf at the time of inoculation. The method enables cultivars or breeding lines of onion species to be reliably screened for resistance to larvae of this pest in a glasshouse so that only the most promising need be tested in the field.     

Influence of Temperature, Wetness Duration, and Leaf Type on the Quantification of Monocyclic Parameters of Bean Rust

Monocyclic parameters of bean rust (Uromyces phaseoli var. typical) were quantified in growth chambers, on rwo bean cultivars for three temperatures (17, 21, and 25 °C), two types of leaves (unifoliolate and trifoiiolate leaves), and nine leaf wetness periods (0, 4, 7, 10, 13, 16, 19, 22, and 25 hrs). The expression of disease was greatly influenced by past-inoculation temperatures. The incubation and latent periods were shortest at 21 °C for both cultivars and leaf types. For both cultivars, trifoiiolate leaves were more susceptible than unifoliolate leaves. A wetness period of at least four hours was required for disease to occur. The maximum disease efficiency for both cultivars occurred with 22 hrs of leaf wetness at 17 °C. The disease efficiencies for temperatures of 17–29 °C and leaf wetness periods of 0–25 hrs were adequately described by a response-surface model. Because of the great influence of temperature and leaf wetness on infection, bean rust is unlikely to occur at high temperatures (> 25°C) and short leaf wetness periods (< 7 hrs).     

Vector relationships and other characteristics of barley yellow striate mosaic virus (BYSMV)

Barley yellow striate mosaic virus (BYSMV) was inoculated by its planthopper vector Laodelphax striatellus (Homoptera, Delphacidae) to 44 species of Gramineae, 26 of which in eight tribes were infected. The virus was not transmitted through wheat seed nor did it infect five dicotyledonous hosts of other rhabdoviruses. The most susceptible species were in the tribes Festuceae and Hordeae. Barley, Bromus spp., oats, Phalaris canariensis, Setaria italica, Sorghum spp., and sweet corn cv. Golden were diagnostic hosts. Electron microscopy of crude sap was also a sensitive diagnostic method. Properties of BYSMV were determined by injecting L. striatellus with crude sap from infected barley. Sap was infectious after 10 min at 50–55 °C but not after 10 min at 60 °C, when diluted with buffer to 10^--² but not to 10^--³, when stored for 2 but not 4 days at 5 °C or when kept for 1 but not 2 days at 22 °C. The planthopper Javesella pellucida was an experimental vector of BYSMV but the virus was not transmitted by the leafhoppers Macrosteles sexnotatus or Psammotettix striatus (Homoptera, Cicadellidae). The latent period of BYSMV in L. striatellus was most commonly 15 or 16 days (minimum, 9 days; maximum, 29 days). The minimum acquisition access period for transmission was between 1 h and 5 h, and the minimum inoculation feeding time was 15 min. After 24 h and 8 day acquisition feeds, 30.4% and 42.8% respectively of L. striatellus transmitted BYSMV. When transferred daily, infective hoppers transmitted virus intermittently. The maximum retention of infectivity by L. striatellus was 36 days. Two of five infective females transmitted BYSMV transovarially. Larvae became infective in the second wk after hatching and transmitted for up to 3 wk.     

Serological studies on cassava latent virus

Particles of cassava latent virus (CLV) were purified by a method that yielded up to 3 mg per 100 g of systemically infected Nicotiana benthamiana leaf. Specific antiserum was prepared and used for enzyme-linked immunosorbent assay (ELISA), which detected purified virus at 5 ng/ml. As estimated by ELISA, CLV antigen reached a greater concentration in leaves of N. benthamiana plants kept at 20–25 °C than in those at 15 °C or 30 °C. CLV was also detected in leaf extracts of naturally infected cassava plants kept at 25 C but its concentration was only 1–7% of that in comparable extracts from N. benthamiana. Staining sections of N. benthamiana leaves with fluorescent antibody indicated that CLV particle antigen accumulates in the nuclei of many phloem cells and of some cells in other tissues. In tests on mosaic-affected cassava plants of Angolan origin, three plants were found in which CLV could not be detected by either ELISA or immunosorbent electron microscopy, or by transmission to indicator plants. This suggests that the mosaic symptoms were caused by a pathogen other than CLV, but no such agent was detected by electron microscopy of leaf extracts. Three kinds of serological test indicated that CLV is related to bean golden mosaic virus. Evidence was also obtained of a distant relationship to beet curly top virus but none was detected to four other geminiviruses.