Genetic basis of resistance to systemic infection by Xanthomonas axonopodis pv. dieffenbachiae in Anthurium

The genetic basis of systemic resistance to bacterial blight disease (blight) of anthurium (Anthurium andraeanum) caused by Xanthomonas axonopodis pv. dieffenbachiae was investigated in progenies of 53 crosses involving 31 parent cultivars using segregation analysis. Inoculation of parents and progenies was achieved by injecting the petiole base of the most recent fully expanded leaf with 100 microl of 10(9) colony forming units per ml of the blight pathogen (strain X4gfp) transformed with the green fluorescent protein (GFP) gene. The time to death and the presence or absence of GFP fluorescence on newly emerging leaves was monitored over a period of 30 weeks after inoculation (WAI), on an individual plant basis. The expected resistance to susceptible ratios based on a digenic model involving two dominant genes, designated A and B, interacting according to a duplicate recessive epistasis model fitted the observed segregation ratios in the crosses. Based on the segregation ratios obtained, the parental cultivars were assigned plausible genotypes. There were significant differences (P < 0.001) in time to death following inoculation between the various genotypic designations. Cultivars with genotypes AABB, AABb, AaBB, and AaBb died within 10 WAI and designated as susceptible; AAbb and aaBB died from 18.8 to 25.6 WAI and were designated as moderately resistant; and Aabb, aaBb, and aabb produced resistant phenotypes. There was also some evidence for dosage effect especially in the highly resistant category. Hence, (AABb = AaBB = AaBb) < (aaBB = AAbb) < Aabb = aaBb = aabb). An approach to fixing resistance to blight in anthurium is discussed.     

Factors influencing Ralstonia pseudosolanacearum infection incidence and disease development in rose plants

Glasshouse experiments were conducted to study infection and disease development in rockwool-grown rose plants inoculated with Ralstonia pseudosolanacearum. A R. pseudosolanacearum strain isolated from rose plants was more aggressive than strains from anthurium or curcuma. The three rose cultivars tested, Avalanche, Red Naomi, and Armando, differed in susceptibility. At 20°C, the rose strain caused hardly any symptoms over a 6-week period, whereas at 28°C typical wilt symptoms were observed within 2 weeks after stem inoculation of Armando, the most susceptible cultivar. Inoculating roots with the rose strain resulted only in weak atypical symptoms. Nevertheless, inoculating roots of cv. Armando at a relatively low inoculum dose of 10⁴ cfu/ml led to high densities in the base of stems in one out of two experiments. R. pseudosolanacearum occasionally spread from stem inoculated plants with symptoms in rockwool slabs. This limited spread resulted in a low infection incidence, and only of plants directly adjacent to the plants with symptoms.     

Genetics of resistance to cotton leaf curl disease in Gossypium hirsutum

Twenty-two cotton varieties were screened for resistance to cotton leaf curl disease (CLCuD), a disease of viral origin, using three procedures: field evaluation, whitefly transmission assay and graft inoculation. Viral infection of cotton varieties was determined by visual symptom assessment as well as dot-blot and multiplex PCR diagnostic techniques. Crosses were made between the most susceptible variety (S-12) and highly resistant varieties (CP-15/2, LRA-5166 and CIM-443). All F₁ plants of these crosses were resistant, showing dominant expression of the resistance as well as the absence of extrachromosomal inheritance. The F₂ plants of the crosses CP-15/2 × S12, LRA-5166 × S-12 and CIM-443 × S12 exhibited a ratio of 13 resistant (symptomless) to three susceptible (with symptoms). Screening of the F₂ generation for virus infection by multiplex PCR further subdivided the resistant class into those exhibiting a high level of resistance (HR; PCR-negative) and those exhibiting resistance (R; symptomless, yet showing virus replication by PCR analysis). Hence, the final ratio was 3:10:3 (HR:resistant:susceptible). The F₃ progeny of susceptible F₂ plants segregated for resistance, indicating the probable presence of a suppressor gene ( S ). These findings are consistent with three genes being involved in G. hirsutum resistance to CLCuD, two for resistance ( R _1CLCuDhir and R _2CLCuDhir ) and a suppressor of resistance ( S _CLCuDhir ).     

Spatial and temporal spread of powdery mildew (Erysiphe pisi) in peas (Pisum sativum) varying in quantitative resistance

A field experiment was conducted to assess the progress in time and spread in space of powdery mildew (caused by Erysiphe pisi ) in pea ( Pisum sativum ) cultivars differing in resistance to the disease. Disease severity (proportion of leaf area infected) was measured in 19 × 23 m plots of cultivars Pania and Bolero (both susceptible) and Quantum (quantitatively resistant). Inoculum on infected plants was introduced into the centre of each plot. Leaves (nodes) were divided into three groups within the canopy (lower, middle, upper) at each assessment because of the large range in disease severity vertically within the plants. Disease severity on leaves at upper nodes was less than 4% until the final assessment 35 days after inoculation. Exponential disease progress curves were fitted to disease severity data from leaves at middle nodes. The mean disease relative growth rate was greater on Quantum than on Pania or Bolero, but it was delayed, resulting in an overall lower disease severity on Quantum. Gompertz growth curves were fitted to disease progress on leaves at lower nodes. Disease progress on Quantum was delayed compared with Pania and Bolero. The average daily rates of increase in disease severity from Gompertz curves did not differ between the cultivars on these leaves. Disease gradients in the plots from the inoculum focus to 12 m were detected at early stages of the epidemic, but the effects of background inoculum inputs and the rate of disease progress meant that these gradients decreased with time as the disease epidemic intensified. Spread was rapid, and there were no statistically significant differences between cultivar isopathic rates (Pania 2.2, Quantum 2.9 and Bolero 4.0 m d⁻¹).     

Resistance of potato cultivars to Synchytrium endobioticum in field and laboratory tests, risk of secondary infection, and implications for phytosanitary regulations

Laboratory (Spieckermann) tests, pot tests and field tests provided concordant evidence for the partial nature of resistance of potatoes to pathotypes 1 (D1) and 6 (O1) of Synchytrium endobioticum . Susceptible potato cultivars produced large warts (> 16 mm in diameter) in Spieckermann tests and had low field resistance levels (1–6). Field-resistant cultivars (levels 7–9) produced small warts or no warts at all in Spieckermann and field tests. In pot tests, at low inoculum levels (1 sporangium per 25 g soil) susceptible cultivars still developed warts, whereas field-resistant ones did not develop any warts below 25 sporangia per g soil. Above 35 sporangia per g soil, 100% disease incidence was observed in susceptible cultivars but only minimal wart development in field-resistant ones. Tests with continuous cultivation of potato cultivars in infected soil during three consecutive years showed that field-resistant cultivars will not support build-up of inoculum in soil. It is concluded that field-resistant cultivars do not create a risk of secondary infection, the criterion given for resistance in EU Directive 69/464/EC.     

Effect of method of inoculation, inoculum density and seedling age at inoculation on the expression of resistance of cocoa (Theobroma cacao L.) to Verticillium dahliae Kleb.

Soil drench and stem puncture inoculation were compared as methods for selecting cocoa cultivars with resistance to Verticillium dahliae. Disease progress was more rapid and induced symptoms were more severe following stem puncture and, under glasshouse conditions, differences between cultivars were detected 15 days after inoculation. Moreover, using stem puncture, inoculum densities of 10⁴ conidia/ml were sufficient to differentiate resistant and susceptible cultivars, whereas with the soil drench method, inoculum densities of 10⁷ conidia/ml were necessary. Although a substantially higher proportion of plants were affected by stem puncture inoculation, the resistance of cultivar Pound-7 remained effective at high inoculum densities of 10⁸ conidia/ml. With either method, older seedlings were more susceptible to V. dahliae than younger ones. However, with stem puncture, 15-day-old seedlings were sufficiently susceptible for a valid disease assessment. In contrast, with soil inoculation, 60-day-old plants were required. In a nursery trial with 15-day-old seedlings, seven cocoa genotypes previously selected as resistant, moderately resistant or susceptible to Verticillium dahliae , on the basis of root inoculation, were ranked in the same order when stem punctured. Stem puncture inoculation of young seedlings is cost-effective in terms of time and space, and is therefore recommended for screening of cocoa for wilt resistance, especially in large-scale breeding programmes.     

Systemic Potato virus YNTN infection and levels of salicylic and gentisic acids in different potato genotypes

Endogenous levels of free and conjugated salicylic (SA) and gentisic (GA) acids, both putative signal molecules in plant defence, were analysed in order to investigate their involvement in the resistance of four potato ( Solanum tuberosum ) genotypes with different susceptibilities to Potato virus Y^NTN (PVY^NTN) infection: the highly susceptible cv. Igor and its extremely resistant transgenic line, the extremely resistant cv. Sante and the tolerant cv. Pentland Squire. The lowest levels of free and conjugated SA were observed in the extremely resistant cv. Sante, while free GA, which was detected in all the other varieties, was absent. The extremely resistant transgenic cv. Igor contained the highest basal total SA level and the lowest level of total GA of all four cultivars. In susceptible cv. Igor, but not in resistant transgenic cv. Igor, a systemic increase of free SA was measured 1 day postinfection (dpi). Even more significant increases of free and conjugated SA and GA were detected 11 dpi when systemic symptoms appeared. In inoculated but not in upper noninoculated leaves of resistant transgenic cv. Igor, significant increase of SA conjugates occurred, but not before 11 dpi. The increase of SA and GA in susceptible cv. Igor could contribute to the general elevated levels of phenolic compounds as a response to stress caused by virus infection. It appears that basal levels of SA and GA do not correlate with resistance to PVY^NTN in potato plants.     

Effect of inoculum type and timing of application of Coniothyrium minitans on Sclerotinia sclerotiorum: control of sclerotinia disease in glasshouse lettuce

The effects of Coniothyrium minitans inoculum quality and an 8-week interval between inoculum application and crop planting on sclerotinia ( Sclerotinia sclerotiorum ) disease in three successive lettuce crops were investigated in a glasshouse trial. Spore suspensions of three isolates of C. minitans (Conio, IVT1 and Contans) applied at 10⁸ CFU m⁻² and a standard Conio maizemeal–perlite application (06 L m⁻², 10¹¹ CFU m⁻²) were assessed for their ability to control S. sclerotiorum . Only the maizemeal–perlite inoculum (isolate Conio) consistently reduced sclerotinia disease. In the third lettuce crop only, isolates IVT1 and Contans formulated by Prophyta and isolate IVT as an oil–water formulation, all applied as spore suspensions, reduced disease at harvest compared with the untreated control. Recovery, viability and C. minitans infection of sclerotia buried during the 8-week period prior to each of the three lettuce crops, and of sclerotia formed on the crop, were tested. Only the maizemeal–perlite inoculum (isolate Conio) reduced the recovery of sclerotia buried in soil for weeks between inoculum application and crop planting, reducing their viability and increasing infection by C. minitans . Eight weeks was sufficient to enable C. minitans to infect sclerotia of S. sclerotiorum , and may account for disease control. After harvest of the second and third crops, maizemeal–perlite treatment (isolate Conio) reduced the number and viability of sclerotia recovered on the soil surface and increased infection by C. minitans compared with spore-suspension treatments. The effect of inoculum concentration and the influence of soil temperature (varying with time of year) on infection of sclerotia by C. minitans are discussed.     

Response of interspecific Brassica juncea/Brassica rapa hybrids and their advanced progenies to Albugo candida (white blister)

Transfer of factors for resistance to white blister disease caused by Albugo candida between Brassica species involving two genotypes each of B. juncea and B. rapa was studied in hybrids. More hybrids were obtained by in vivo than in vitro techniques, although an in vitro phase was a prerequisite for the establishment of in vivo hybrids. Hybrids were identified by PCR-based inter-simple sequence repeat (ISSR) markers with both male and female species-specific bands being identified. There was a positive correlation between disease severity and number of days after sowing ( r  > 0·93), the highest being towards pod formation and plant maturity at 110 days after sowing. The plants from F₂ and BC₁ progeny showed higher resistance to A. candida than either of the parents. Plants of B. juncea and B. rapa with high field resistance (disease index < 1·0) were selected from BC₂ and F₂BC₁ generations. The frequency of plants classified as resistant in BC₂ progeny ranged from 4·5 to 39·0% in cross-combinations involving B. juncea genotypes as female parent, compared with 100% in the reciprocal cross involving B. rapa as female parent.     

Modelling effects of vector acquisition threshold on disease progression in a perennial crop following deployment of a partially resistant variety

Deployment of resistant varieties is a key strategy to mitigating economic losses due to arthropod‐transmitted plant pathogens of perennial crops. In many cases, the best available resistant traits for introgression confer only partial resistance. Plants displaying partial resistance have lower pathogen titres than susceptible counterparts, but remain hosts for the pathogen. As partially resistant varieties maintain yield after infection, infected plants are unlikely to be rogued (i.e. removed). Accordingly, there is a risk that partially resistant plants could serve as a source of inoculum for pathogen spread to susceptible plants. Here, a mathematical model that tracked spread of an arthropod‐transmitted pathogen in a plant population consisting of susceptible and partially resistant plants was used to identify a threshold acquisition rate from partially resistant plants that resulted in limited spread of the pathogen from partially resistant plants to susceptible plants. The acquisition threshold from partially resistant plants varied with parameters influenced by disease management decisions such as number of vectors per plant, vector turnover, replacement of susceptible plants, and proportion of plants that were partially resistant. In model simulations, effects of deploying a partially resistant variety on disease incidence in a susceptible variety depended on the extent to which pathogen spread among susceptible plants was suppressed and acquisition rates from partially resistant plants. Collectively, the results indicate that risk of partially resistant plants serving as inoculum sources could be assessed prior to deployment, thereby enabling design of complementary disease management tactics to minimize economic losses in susceptible varieties following deployment.     

Association of induction of protease and chitinase in chickpea roots with resistance to Fusarium oxysporum f.sp. ciceri

To improve the breeding of chickpea varieties with resistance to Fusarium wilt, an attempt was made to analyse the biochemical basis of disease resistance by measuring levels of chitinase, β-1,3-glucanase, protease and proteinase inhibitor activities in dry and soaked seeds and in root and shoot tissues of wilt-resistant and wilt-susceptible cultivars. Marginal variation was observed in the levels of the candidate proteins in dry or soaked seeds. Chitinase activity was higher in roots than in shoots or cotyledons. No proteinase inhibitor activity was detected in root and shoot tissue of any of the cultivars. When the levels of these proteins were analysed in resistant (Vijay) and susceptible (JG-62) cultivars during development of wilt by growing plants in soil infested with F. oxysporum f.sp. ciceri , race 1, both cultivars showed induction of chitinase activity in the roots. However, the induced activity in JG-62 (3.82 U g⁻¹) was equivalent to the constitutive level in Vijay (3.90 U g⁻¹) and much lower than that induced in Vijay (5.18 U g⁻¹). Induction of protease activity was observed only in root extracts of Vijay when challenged by the pathogen. The root extract of Vijay showed in vitro antifungal activity in a plate assay. Simultaneous induction of proteolytic and chitinolytic activities specifically in the resistant cultivar was correlated with antifungal properties of root extracts effective in conferring resistance.     

Recovery of Young Olive Trees from <Emphasis Type="Italic">Verticillium dahliae</Emphasis>

Natural recovery from wilt disease symptoms was evaluated in young olive trees root dip inoculated with Verticillium dahliae in a growth chamber over a 12 week period and, later on, when the trees were transplanted in a V. dahliae-free soil in a lathhouse during a period of 127 weeks. Recovery in an individual tree was considered when a plant showed symptom remission after having reached a maximum value of symptom severity. Recovery accounted for 53% of 464 trees that showed wilt symptoms during observations in the two environments. The remaining trees died. Recurrent wilt symptoms were not observed in recovered trees, and recovery was usually accompanied by the production of new green tissues. Recovery was clearly higher in trees inoculated with a non-defoliating (ND) isolate (86.4%) of the pathogen than in those inoculated with a defoliating (D) isolate (23.9%). The percentage of recovery and the level of resistance were significantly correlated. Recovery accounted for 92.1% of the cases in resistant and moderately susceptible cultivars, reaching 100% in plants inoculated with the ND isolate (Table 2); meanwhile it was three times lower (30.1% of the plants) in susceptible and extremely susceptible diseased trees. In the lathhouse, periodical tissue isolations for monitoring the progress of infections over a period of 127 weeks in recovered trees, showed that the pathogen could only be isolated from trees 19 weeks after inoculation. Pathogen isolation was significantly higher from susceptible and extremely susceptible cultivars (84.6%) than from resistant and moderately susceptible ones (33.3%). Results showed that if a tree overcomes infection by pathogen from a single inoculation, and it is able to begin a recovery process, it will not express wilt symptoms again in a pathogen-free environment. The pathogen remained inactive or dead over time in recovered trees. Thus, new infections from rootlets would be necessary for new symptom expression. Recovery from Verticillium wilt is an important natural mechanism that occurs in a high percentage of infected olive trees, and can complement the resistance of the cultivar, particularly in conditions of low inoculum densities of low virulence isolates of the pathogen in the soil.     

The Effect of Different Levels of Beet Cyst Nematodes (Heterodera schachtii) and Beet Necrotic Yellow Vein Virus (BNYVV) on Single and Double Resistant Sugar Beet Cultivars

Beet cyst nematode-resistant cultivars, which were introduced recently, originated from the homozygous inbred line B883. This translocation product was unstable and the transmission of resistance when crossed with a susceptible cultivar did not exceed 94%. Tests with the resistant cultivars in climate cabinets showed a wide variety of resistance against Heterodera schachtii and beet necrotic yellow vein virus (BNYVV), expressed as average numbers of infective units per plant and percentages of resistant plants. In a series of field trials at different levels of infection of H. schachtii, their multiplication rates on all resistant cultivars depended on the initial density, which was caused by the presence of small numbers of susceptible plants. Since tolerance to wilting was also incorporated in B883, reasonable yields were obtained in the presence of H. schachtii. However, at increasing initial densities of H. schachtii, yields decreased considerably, since penetrating juveniles cause a hypersensitivity reaction in resistant plants. Based upon the results of three series of field trials, it was concluded that resistant cultivars should preferably be applied at population densities between 500 and 2000 eggs and juveniles of H. schachtii per 100ml of soil. Cultivars with double resistance against H. schachtii and BNYVV behaved like those with H. schachtii resistance in soils infected with beet cyst nematodes, but not with BNYVV. In soils with a combined infection of H. schachtii and BNYVV double resistant cultivars were far superior to single resistant ones, since damage caused by BNYVV was far more serious than damage caused by H. schachtii. No substantial interaction between soil pathogens nor types of resistance could be detected.     

Colonization and sporulation of Peronospora viciae on cultivars of Pisum sativum

The behaviour of Peronospora viciae was examined on four pea ( Pisum sativum ) cultivars (Maro, Superb, Victory Freezer and Surprise) which appeared to exhibit varying degrees of resistance to this pathogen. Penetration into all cultivars was similar and mainly by direct entry through the cuticle. The pathogen grew in a similar manner and at a similar rate in all the cultivars during the early stages (<96 h) of tissue colonization. As it approached the onset of sporulation its growth slowed down in the more resistant cultivars but in no instance were any hypersensitive or other host cell reactions observed. Greater differences between the cultivars were seen in the extent of sporulation, with the least resistant cultivar (Superb) bearing the largest number of sporangiospores. There was much less sporulation on the most resistant cultivar (Maro) and the process was more dependent on temperature than it was on Superb. The development of mature plant resistance occurred in a similar way and on a similar time-scale in all cultivars. The youngest leaflets on plants which were less than 8 weeks old at inoculation showed the least resistance to infection and colonization.     

Coinfection of soybean plants with Phytophthora sojae and soybean cyst nematode does not alter the efficacy of resistance genes

The soybean cyst nematode (SCN) Heterodera glycines and the oomycete Phytophthora sojae are among the most damaging pathogens of soybean worldwide. Resistant cultivars are commonly used to manage these diseases. As it is known that the presence of SCN can facilitate the development of other pathogens, it is important to verify if there is a synergistic activity between SCN and P. sojae. The purpose of this study was to evaluate a possible interaction on susceptible and resistant soybean lines. The plants were inoculated with one or both organisms at different stages (5 or 10 days old). Two levels of SCN inoculum (2,000 and 10,000 eggs/plant) and different timing between SCN and P. sojae inoculation (2, 5, or 8 days) were compared. The results on 5-day-old plants showed that SCN did not influence P. sojae development. The resistant cultivar to P. sojae remained effective (0% mortality) and susceptible cultivars exhibited high mortality (100%) in the presence or absence of SCN. Experiments on 10-day-old plants showed that SCN resistance was not affected by the presence of P. sojae. SCN inoculum density and timing of P. sojae infection did not affect the virulence of these pathogens and the efficacy of resistance genes. However, the number of SCN cysts was decreased by more than 50% (p < .001) when P. sojae was coinfesting the susceptible cultivar. This suggests that P. sojae might indirectly influence SCN development by reducing the root mass. This study confirmed that resistant cultivars remain a valid option for the management of P. sojae and SCN.     

Resistance in water yam (Dioscorea alata) cultivars in the French West Indies to anthracnose disease based on tissue culture-derived whole-plant assay

Reactions of 60 water yam ( Dioscorea alata ) cultivars to three isolates of the yam anthracnose fungal pathogen ( Colletotrichum gloeosporioides ) were evaluated using tissue culture-derived whole-plant assay. Three disease parameters: single score on a scale of 0–6 at the seventh day after inoculation (SD7); area under the disease progress curve (AUDPC); and disease progress rate ( R _d) were compared, and cultivars were classified into disease-response groups using a rank-sum method based on AUDPC scores for the two most virulent isolates. A wide range of variation in resistance of the D. alata cultivars, and significant effects of pathogen isolate and isolate–cultivar interactions, were observed for all disease parameters. The three disease parameters were positively correlated; however, four cultivars showed great dispersions from the regression lines for comparisons of SD7 with the multiple assessments based AUDPC and R _d. The 60 cultivars were separated into resistant ( n  = 12), moderately resistant ( n  = 19), moderately susceptible ( n  = 18) and susceptible ( n  = 11) groups. The potential of the tissue culture-derived whole-plant assay to resistance breeding programmes and further understanding of the yam anthracnose pathosystem is discussed.     

Modelling different cultivation and herbicide strategies for their effect on herbicide resistance in Alopecurus myosuroides

A single dominant mutation conferring resistance to aryloxyphenoxypropionate (AOPP) and cyclohexanedione (CHD) herbicides was incorporated into a quantitative model for the population development of Alopecurus myosuroide s Huds. The model predicts that from an initial seedbank of 100 seed m^–2, 10^–6 of which mutate to resistance each generation, and annual use of AOPP/CHD herbicides which kill 90% of susceptible but no resistant plants, a threshold of 10 plants m^–2 surviving herbicides ('field resistance') will develop: in 9–10 years if all tillage is by tine cultivation to 10 cm deep; after 28–30 years of annual ploughing; in 12 years if tine cultivations are interspersed with ploughing once every 4 years. If AOPP/CHD herbicides are alternated with herbicides with different modes of action, outcomes depend on the annual kill rate: with 95% kill (of susceptible plants by AOPP/CHDs and all plants by alternative herbicides) and tine cultivation, field resistance develops in 22 years; however, resistance can be delayed for 45 years if AOPP/CHDs are rotated with two additional herbicides, each with a different mode of action. The model predictions on the number of years required for field resistance to develop are not highly sensitive to the density of the seedbank or the initial frequency of resistance.     

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.     

Selection for resistance to the herbicide imazethapyr in somaclones of soyabean

Cultivars of soyabean [ Glycine max (L.) Merr.] resistant to the herbicide imazethapyr were identified by suspending the roots of 5-day-old seedlings in nutrient culture containing 2.5 mg a.i. L^–1 imazethapyr and then comparing the inhibitory effect on root length and shoot dry weight. The four most resistant cultivars were subsequently screened as regenerating tissue cultures in a medium containing 2.0 mg a.i. L^–l imazethapyr to select somaclonal cells with increased resistance. Surviving portions of cultures were regenerated to give shoots, the plants isolated, allowed to flower and seed. These progeny were then used for further seed multiplication and seedlings from this latter generation were exposed to imazethapyr in vivo and callus and cell suspension cultures derived from these seedlings were exposed to imazethapyr in vitro . A reduction in the inhibitory effect of the imazethapyr was noted in the somaclone seedlings and tissue cultures. However, measurement of acetolactate synthase (ALS) activity showed no differences among the parent cultivars and in the selected somaclones in this trait.     

Effects of inoculum concentration, wetness duration and plant age on development of early blight (Alternaria solani) and on shedding of leaves in tomato plants

Effects of inoculum concentration, wetness duration and plant age on the development of tomato early blight were evaluated in relation to host susceptibility under controlled environmental conditions. The main effect of early blight was premature defoliation, which was linearly related to the percentage of leaf area showing symptoms. As ln(inoculum concentration, conidia mL⁻¹) increased from 6·2 to 11·5, the percentages of leaf area affected and of defoliation increased linearly. Four h of leaf wetness after inoculation were sufficient to initiate the disease on plants of hybrid Skala RZ but not on those of cv. Rio Rojo, for which at least 6 h leaf wetness were needed. As wetness duration increased up to 24 h, there was an increase in the percentage leaf area showing symptoms and in the percentage of defoliation, but thereafter there was no significant increase in either parameter. Tomato plants were susceptible to Alternaria solani at all growth stages, but susceptibility increased as plants matured. There were no significant differences in susceptibility between tomato cultivars and hybrids.