Factors influencing the production of basidiocarps and the deposition and germination of basidiospores of Crinipellis perniciosa, the causal fungus of witches' broom on cocoa (Theobroma cacao)

The production of basidiocarps by Crinipellis perniciosa on detached, dead witches'brooms from cocoa was assessed in relation to temperature, light, cocoa clone, age of broom and type of tissue, in cabinets with a daily cycle of 8 h wet and 16 h dry. More basidiocarps formed and matured at 20–25°C than at 25–30°C. In the latter regime the pilei were smaller and white, instead of the usual crimson colour, and the stipes were longer. No basidiocarps formed at 30–35°C. At 20–25°C. more basidiocarps formed and matured with light at 100 μE m^-2 s^-1 during the wet period than at 10 μE m^-2 s ^-1. Only one basidiocarp and five primordia developed on 20 brooms kept in the dark. Brooms from 10 cocoa clones at Pichilingue. Ecuador, differed in basidiocarp productivity. most basidiocarps forming on brooms from Seavina and least on ICS clones. The numbers of basidiocarps produced on brooms aged 1.2.3 or 4 months when detached from cocoa trees were similar but time to initiation of the first primordium differed considerably. More basidiocarps formed at nodes than internodes.
The discharge of basidiospores was optimal at 20–25°C and 80% RH: germination was optimal in water agar films. Neither process was dependent on light.     

Real-time vision-based detection of Rumex obtusifolius in grassland

Rumex obtusifolius is a common grassland weed that is hard to control in a non-chemical way. The objective of our research was to automate the detection of R. obtusifolius as a step towards fully automated mechanical control of the weed. We have developed a vision-based system that uses textural analysis to detect R. obtusifolius against a grass background. Image sections are classified as grass or weed using 2-D Fourier analysis. We conducted two experiments. In the first (laboratory) experiment, we collected 28 images containing R. obtusifolius and 28 images containing only grass. Between 23 and 25 of 28 images were correctly classified (82–89%) as showing R. obtusifolius ; all grass images were correctly classified as such. In the second (field) experiment, a self-propelled platform was used to obtain five sequences of images of R. obtusifolius plants. We used the parameters that gave the best classification results in the first experiment. We found, after changing one of the algorithm's parameters in response to prevailing light conditions, that we were able to detect R. obtusifolius in each image of each sequence. The algorithm scans a ground area of 1.5 m² in 30 ms. We conclude that the algorithm developed is sufficiently fast and robust to eventually serve as a basis for a practical robot to detect and control R. obtusifolius in grassland.     

Effects of light and mechanical stimulation on coiling and prehaustoria formation in Cuscuta spp.

Irradiation of 4-day-old dark-grown seedlings of Cuscuta campestris Yuncker and Cusctira indecara Choisy with I min of red light stimulated coiling and prehaustoria formation in excised upper 4-cm segments of seedlings during a 48-h dark period in which zeatin was applied. Coiling and prehaustoria development were completely reversed when irradiated with 2 min of far-red light applied directly after red light. When deetiolated segments were subjected to prolonged exposure of pure blue light, or mixtures of ultraviolet-A/far-red, blue/far-red or red/far-red light, coiling and prehaustona development were induced. Blue light at photon fluxes lower than 1 μmol m^-2 s^-1 did not promote coiling and formation of prehaustoria. Mechanical stimulation caused tendrils to coil and form prehaustona in biue and blue/far-red light, but not in darkness. Zeatin substituted for high irradiance blue light and mechanical stimulation caused coiling and prehaustoria formation.     

A qualitative quick-test for detection of herbicide resistance to tribenuron-methyl in Papaver rhoeas

A qualitative seed-based method useful for the detection of resistance to the herbicide tribenuron-methyl in Papaver rhoeas L. is described. Seeds were germinated on 35 mL of a 1.3% agar medium containing 2 g KNO₃ L^–1 in 8.5 cm Petri dishes in a growth chamber under 20 μmol s^–1 m^–2 of fluorescent light. When 0.24 μM tribenuron-methyl or more was added, growth in susceptible plants stopped after the cotyledon stage and they turned chlorotic. The resistant plants continued developing new leaves. The same effect was achieved when 0.2 g gibberellin (GA₃) L^–1 and 7.68 μM tribenuron-methyl or 0.5 g GA₃ L^–1 and 61.44 μM tribenuron-methyl were added. Germination percentage rose with gibberellin in the presence or absence of the herbicide. Plants developed rapidly, with only about 14 d needed to finish the test but sometimes root growth was reduced because of the addition of gibberellin. In the absence of gibberellin but in the presence of the herbicide, plants grew more slowly and developed smaller leaves with a 17-d evaluation period requirement. The test was validated with pot experiments in a greenhouse and also with field trials. The best combination was found to be 0.2 g GA₃ L^–1 and 7.68 μM tribenuron-methyl, assuring homogenous germination and testing of dormant seeds but avoiding root inhibition associated with too much gibberellin.     

Light, temperature and burial depth effects on Rumex obtusifolius seed germination and emergence

Trials were carried out to investigate the effects of light and temperature on germination of Rumex obtusifolius L. After several months of storage, seeds gradually lost dormancy and became photosensitive. Thermal optima for germination were between 20 °C and 25 °C in light or in darkness. At lower temperatures there was a greater demand for light, so that the greatest differences in germination percentage (between low and high temperatures) were found within the 10–15 °C temperature range. The calculated thermal minima ( x -intercept method) in light and darkness were 8.3 °C and 6.1 °C respectively. Daily temperature fluctuation increased germination even after seed irradiation with far-red light, suggesting a lower demand for the far-red-absorbing form of phytochrome. Seed burial inhibited germination in proportion to depth; however, germination inhibition was independent of seed phytochrome photo-equilibrium, which had been diversified by seed pretreatment with light. Seedlings did not emerge when seeds were buried >8 cm deep. Recovery of ungerminated seeds showed that excessive burial did not impede seedling emergence but rather prevented seed germination. However, this induction of dormancy was lost once germination processes were activated (24–48 h at 20 °C) that made germination irreversible. Temperature was also involved in inhibition, and low temperature (<15 °C) induced the least inhibition. This is discussed in terms of processes of respiration and fermentation in buried seeds.     

Growth and photosynthesis of four invasive aquatic plant species in Europe

Crassula helmsii , Hydrocotyle ranunculoides , Ludwigia grandiflora and Myriophyllum aquaticum are four well known invasive aquatic plants in European waters. In this study, plant growth at different nutrient availabilities, regeneration capacity and photosynthesis were investigated. Results show high relative growth rates (RGR) of the species of up to 0.132 ± 0.008 g g⁻¹ dry weight (dw) day⁻¹ ( H. ranunculoides ) and a significant increase in RGR with increasing nutrient availability. All species show a high regeneration capacity and the ability to form new shoots from single nodes, even though it differs between the species. Ludwigia grandiflora and M. aquaticum also show regeneration from single leaves. Species differed in maximal amounts, and in temperature and light optima of net assimilation rates: H. ranunculoides leaves reach maximum photosynthetic rates of up to 3500 μmol CO₂ × h⁻¹ g⁻¹ dw, L. grandiflora (leaves) up to 2200 μmol CO₂ × h⁻¹ g⁻¹ dw, M. aquaticum (shoots) 400   μmol CO₂ × h⁻¹ g⁻¹ dw and C. helmsii (shoots) up to 200 μmol CO₂ × h⁻¹ g⁻¹ dw. Hydrocotyle ranunculoides , L. grandiflora and M. aquaticum preferred high light intensity and high temperatures, whilst C. helmsii was negatively affected by intense sunlight. Summarising, it can be assumed that at least H. ranunculoides , L. grandiflora and M. aquaticum can grow well under current and likely future central European climate conditions.     

Striga asiatica seed conditioning and 1-aminocyclopropane-1-carboxylate oxidase activity

Conditioned seeds of Striga asiatica (L.) Kuntze release ethylene, which elicits germination. We investigated the activity of 1-aminocyclopropane-1-carboxylate (ACC) oxidase and respiration during conditioning. Seeds incubated in vivo with ACC, the substrate for ACC oxidase, produced negligible ethylene at the beginning of conditioning or if they were dormant (i.e. would not germinate after conditioning and treatment with stimulant). Non-dormant seeds produced 3000 ηL of ethylene/600 seeds/24 h after 12 days of conditioning. In vitro ACC oxidase activity at day 0 of conditioning produced 500 ηL of ethylene/μg protein/h and 8000 ηL of ethylene/μg protein/h after 12 days of conditioning. Incubation of seeds in strigol before protein extraction did not enhance enzyme activity. Seeds released 4000 μL/L CO₂ in the first 24 h of conditioning, with the rate increasing to 15 000 μL/L/24 h on day 4 and then remaining roughly unchanged. Maximum in vitro activity of ACC oxidase required ACC, catalase, O₂, Fe²⁺, ascorbate and CO₂. In vivo activity of ACC oxidase required ACC and/or germination stimulant(s), suggesting that stimulants may be involved in providing substrates for the ACC oxidase. No difference was observed in the separation of extracted proteins, which suggests that ACC oxidase is activated during conditioning, perhaps as a result of changes in co-factor concentration. Application of these findings to Striga control is discussed.     

Invasiveness evaluation of fireweed (Crassocephalum crepidioides) based on its seed germination features

In China, fireweed ( Crassocephalum crepioides ) is listed as an invasive plant that is also cultivated as a vegetable. To gain a clearer understanding of its invasiveness and rapid spread, we evaluated its seed dispersal ability, and the influences of light, temperature, pH, NaCl stress, moisture content, and storage periods on its seed germination. Its seed dispersal ability is limited. The seed germination of fireweed is inhibited by darkness, temperatures <10°C or >35°C, and a NaCl solution with a concentration >0.15 mol L⁻¹. The optimal conditions under which nearly all the seeds could germinate are light, with temperatures from 20 to 30°C, and a neutral soil with 40% moisture content. The seeds of fireweed have no apparent dormancy and retain a high viability after room storage for 10 months. Fireweed only has a moderate invasive capacity and its wide distribution in China possibly correlates with its cultivation.     

Soil potassium supply and Rumex obtusifolius and Rumex crispus abundance in silage and grazed grassland swards

Three experiments investigating factors influencing the abundance of Rumex spp. (docks) in silage and grazed grassland swards are presented. In Experiment 1, Rumex obtusifolius plants were sown with perennial ryegrass and white clover in pots in March and harvested at either 5- or 10-week intervals between June and October. The 10-week harvest interval increased root dry-matter production of R. obtusifolius compared with the 5-week interval; herbage (above-ground material) production was not significantly affected. In Experiment 2, R. obtusifolius and Rumex crispus population densities in grassland swards were correlated with soil P, K and Mg concentrations, and soil pH. In general, silage swards contained higher population densities than grazed swards. There were significant positive correlations between soil K concentrations and abundance of Rumex spp. in grazed swards and in silage swards. In Experiment 3, R. obtusifolius was sown with perennial ryegrass in pots in March. Treatments consisted of nine rates of K fertilization ranging between the equivalent of 0 and 600 kg K ha⁻¹ year⁻¹. Herbage was harvested at regular intervals (4–6 weeks except during the winter) until May of the following year. In general, perennial ryegrass dry-matter yields were not greatly affected by soil K, whereas limited soil K supply tended to reduce dry-matter production of R. obtusifolius . It is possible that maintenance of moderate soil K concentrations may play a role in limiting abundance of Rumex spp. in grassland.     

Antifungal activity of bergenin, a constituent of Flueggea microcarpa

The antifungal activity of bergenin against some plant pathogenic fungi, namely, Alternaria alternata , A. brassicae , A. carthami , Fusarium udum , F. oxysporum f.sp. ciceri , Curvularia lunata and Erysiphe pisi , was studied. Bergenin as its monosodium salt was effective against all the fungi and the effective dose for complete inhibition of spore germination varied from 15 μg mL⁻¹ for F. udum to 125 μg mL⁻¹ for E. pisi . Experiments on the effect of bergenin on powdery mildew development under glasshouse conditions revealed that it can control powdery mildew of pea at 2000 μg mL⁻¹ by postinoculation treatment, the results being comparable with those of carbendazim (1000 μg mL⁻¹) and wettable sulfur (2000 μg mL⁻¹). It affected hyphal elongation and the number of primary and secondary branches.     

Effect of fungal toxins on germination of Striga hermonthica seeds

Fourteen fungal toxins were assayed in vitro to evaluate their effect on seed germination of the parasitic weed Striga hermonthica . Among them, T-2 toxin proved to be the most active, being able to inhibit 100% seed germination at 10⁻⁵ M, and being still active when tested at a concentration of 10⁻⁷ M (19% inhibition). Deoxinivalenol was also very active, causing 100% and 69% reduction in germination when assayed at 10⁻⁴ and 10⁻⁵ M respectively. Cytochalasin E, tenuazonic acid, fumonisin B₁, enniatin and nivalenol were shown to have an inhibitory effect of around 50% at 10⁻⁴ M, whereas other toxins had lower or no activity. The high activity shown by some fungal toxins suggests that they may have potential for use as more natural and safe herbicides to suppress parasite seed germination.     

Possible involvement of gibberellins and ethylene in Orobanche ramosa germination

Summary Six inhibitors acting at different stages of gibberellin biosynthesis, applied during conditioning of Orobanche ramosa seeds, reduced subsequent germination in the presence of GR₂₄ (a strigol analogue). Ethylene seems to be involved in the induction of germination of conditioned seeds by GR₂₄, as inhibitors of its synthesis or action, applied to conditioned seeds, also strongly reduced induction of germination by GR₂₄. Exogenous ethylene did not induce germination of conditioned seeds, but 2-chloroethylphosphonic acid was able to do so. When inhibitors of gibberellin biosynthesis were applied to conditioned seeds in the presence of GR₂₄, they inhibited germination. These same inhibitors also strongly inhibited germination of conditioned Striga hermonthica seeds in response to GR₂₄; this inhibitory effect was reversed by the addition of 2-chloroethylphosphonic acid. The effect of these inhibitors on S. hermonthica , in which ethylene is a necessary mediator of germination induction by GR₂₄, strongly suggests that ethylene synthesis is also required for the induction of O. ramosa seed germination by GR₂₄. These growth regulators, which inhibit the two steps of germination in O. ramosa , could be useful for the development of methods for early season control of this parasite.     

Effects of humidity, leaf wetness, temperature and light on conidial production by Phaeoisariopsis personata on groundnut

Controlled-environment studies of conidial production by Phaeoisariopsis personata on groundnut are described. With constant relative humidity (RH), conidia were only produced above a threshold (94·5% RH) and there was a linear increase between 94·5% RH and 100% RH. Conidial production was less with continuous leaf wetness (resembling heavy dew) than with continuous 98–99% RH, but it was similar with intermittent leaf wetness and intermittent 98–99% RH (8 h at 70% RH each day). With alternate high (≥97% RH) and low humidity, daily conidial production depended both on the duration of high RH and on the low RH value. With 99% RH at night (12 h), night-time conidial production decreased with the previous daytime RH. After conidial production had started, small numbers of conidia were produced even when the RH was well below the threshold (94·5%). Conidia were produced in continuous light when the photon flux density was 2 μmol/m²/s, but production was completely inhibited with 60 μmol/m²/s. With constant RH, more conidia were produced with a 12 h photoperiod than in continuous darkness. However, more than 75% of the conidia were produced in the dark. With continuous darkness, more conidia were produced during the night (18.00–06.00 h) than during the day, but this biological rhythm was overcome with a (light-night)/(dark-day) regime. With constant 98–99% RH there was a linear increase in conidial production with temperature between 10 and 28°C, and virtually no conidia were produced at 33°C. The daily production of conidia increased with time for 2 to 6 days, depending on the treatment.     

Variation in seed dormancy and light sensitivity in Alopecurus myosuroides and Apera spica-venti

Variation in seed dormancy and light sensitivity was studied in Alopecurus myosuroides and Apera spica-venti . Seeds were collected from different populations, at different dates and from plants emerging in autumn or spring, and used in four experiments. In the first experiment, initial dormancy was investigated in light and darkness. In Expt 2, buried seeds were exhumed on 16 occasions, from September 1997 to March 2000, and germinated in light, in darkness and after a 5-s light exposure. In Expt 3, emergence was recorded for seeds sown in pots outdoors. In Expt 4, stratified seeds of A. myosuroides only were exposed to photon irradiance ranging from 0.1 to 25 600 μmol m⁻². Variation was high among seed collections, but both species showed winter annual dormancy patterns. Apera spica-venti germinated to high percentages in autumn but negligibly in spring. Alopecurus myosuroides germinated less in spring when tested in darkness and after a short light exposure and emerged poorly in spring, which reflected photo-desensitisation during cold stratification. We conclude that the peak of emergence in A. myosuroides , and to some extent in A. spica-venti , is largely regulated by exposure to light interacting with low-level dormancy. This offers valuable information regarding optimal timing of weed control measures.     

Seasonal changes in the germination responses of buried Lamium amplexicaule seeds

Spring-produced seeds of Lamium amplexicaule L. were buried in pots of soil in an unheated glasshouse in June 1978, and at 1–2-month intervals, for 27 months, they were exhumed and tested for germination in light and darkness at temperatures simulating those in the habitat from early spring to late autumn. Freshly-matured seeds were dormant, but by autumn 85% or more germinated in light at 15/6, 20/10, 25/15 and 30/15°C but only 7% or less in darkness. During late autumn and winter germination in light decreased at 25/15 and 30/15 °C but not at 15/6 and 20/10 °C, and germination in darkness increased at 15/6 and 20/10 °C. During late winter and early spring germination in light at 15/6 and 20/10 °C decreased, and seeds lost the ability to germinate in darkness. By the second autumn of burial, seeds germinated to near 100% in light at 15/6 to 30/15 °C and to 10–25% in darkness at 15/6 and 20/10 °C. The cycle of germination responses was repeated during the second winter and spring and the third summer of burial. Autumn-produced seeds were dormant when buried in November 1979, but by spring they germinated to 81 and 36% at 15/6 and 20/10 °C, respectively, in light. These seeds afterripened further during summer. The consequence of seasonal changes in germination responses is that (1) seeds can germinate in the habitat in late summer, autumn and spring but not in early- to mid-summer or in late autumn and winter and (2) during both germination seasons, seeds produced during the previous spring(s) and/or autumn(s) can germinate.     

Weeds in slash-and-burn rice fields in northern Laos

Slash-and-burn farmers in northern Laos consider weeds, insufficient rainfall and rodent damage as the most important constraints to upland rice ( Oryza sativa L.) production. Labour inputs of 140-190 days ha^-1 for weed control result in low labour productivity. Average weed cover observed in rice fields was 5.6, 4.1, 2.1, 1.7, 0.7 and 0.7 cm m^-1 (transect) for Chromolaena odorata (L.) King and Robinson, Ageratum conyzoides L., Commelina spp., Lygodium flexuosum (L.) Sw., Panicum trichoides Sw. and Corchorus spp, respectively. Chromoiaena odorata , which was introduced in the 1930s, has become the main fallow species and is considered a desirable fallow plant by most farmers. Average fallow periods reported for the 1950s, 1970s and 1992 were 38, 20 and 5 years respectively. Reduced fallow periods in the last decades have re-suited in a marked increase in weeding requirements. Above-ground biomass for rice stem, herbaceous plants and trees after rice harvest was 168, 67 and 60gm^-2 in 1991 and 115, 43 and 24 gm^-2 in 1992. Weeding at 14-day intervals did not increase rice grain yield.     

Effects of maternal light environment on germination and morphological characteristics of Sicyos deppei seeds

The effect was studied of sunlight and far-red (FR) light during seed development, on seed quality and germination of Sicyos deppei G. Don. Seeds exposed to FR during development were lighter in colour and their weight, size and water content were significantly lower. Less than 10% of non-scarified freshly harvested seeds germinated. Scarified, freshly harvested seeds developed under sunlight had a partially negative photoblastic response; both red (R) and FR light inhibited germination. The highest and fastest germination occurred in darkness, probably due to the effect of the high photon flux densities on the phytochrome during seed development. Scarified seeds ripened under FR light, germinated well in FR light and in darkness, but R light inhibited germination. After 6 months of storage, the permeability of S. deppei seeds increased, the partially negative photoblastic response was lost and germination of scarified seeds increased. Specifically, in seeds developed under FR, germination in darkness was faster than for the other light treatments, but slow in darkness for seeds developed under sunlight. The physiological and morphological heteroblastic responses in S. deppei probably extend its seed germination and seedling recruitment periods.     

The impermeability of powdery mildew conidia and their germination in arid environments

Conidia of Erysiphe graminis formed germ hyphae while floating on aqueous solutions of glutaraldehyde (0.25 M), formaldehyde (1 M), NaF (0.1 M), HgCl (0.05 M) and HgCl₂. (0.1 M). Inhibition of germination ranged from 60.1% (NaF) to 88.8% (HgCl₂). In an atmosphere of HCN gas (5 mM) inhibition was 72.8%. The detergents Triton X-100. sodium dodecylsulphate and cetyl trimethylammonium bromide at 10 g 1^-1 caused approximately 90% inhibition but at lower concentrations Triton X-100 was the most inhibitory, both to germination and germ hypha elongation. Results of simultaneous treatment with HCN and Triton X-100 suggested that their actions were independent but comparisons of LD 50 for glutaraldehyde and Triton X-100 used separately and together indicated antagonism.     

Relationships between seed germination, fumonisin content, and Fusarium verticillioides infection in selected maize samples from different regions of Costa Rica

Thirty-five maize seed samples from three geographic regions of Costa Rica were analysed for fumonisin content, germinability, and Fusarium verticillioides infection with and without surface disinfection. The concentration of fumonisins in the maize samples varied from 4 ng g⁻¹ to 16 000 ng g⁻¹ with an all over-all average of 2500 ng g⁻¹. There was a significant difference in fumonisin content between samples from Alajuela and Guanacaste. Germination of the seed samples ranged from 12% to 98% with significant differences between regions. F. verticillioides infection was 12–98% and 13–97% for surface-disinfected and nondisinfected seeds, respectively. There was a significant negative correlation ( r  = -0.52) between fumonisin content and seed germination, but no significant correlation was found between fumonisin content and F. verticillioides infection, or between F. verticillioides infection and seed germination. Most of the high fumonisin seed samples had some mechanical or insect damage. Whether or not the fumonisins had a direct effect on germination was not further established. It is concluded that the large differences in fumonisin content of maize seeds within and between regions are primarily caused by differences in seed quality, genetic diversity of F. verticillioides strains in natural populations, climatic differences between regions, and varietal differences. Some of the fumonisin levels found in this study coincide with levels associated with risks to humans and animals in other countries.     

Effect of the fungal pathogen Bipolaris sorokiniana toxin prehelminthosporol on barley root plasma membrane vesicles

The effects of the phytotoxin prehelminthosporol from the fungal pathogen Bipolaris sorokiniana on the activities of the plasma membrane H⁺-ATPase, Ca²⁺- ATPase, and 1, 3-β-glucan synthase in barley roots were investigated in vitro. Plasma membranes were isolated by aqueous polymer two-phase partitioning. Proton pumping by the H⁺-ATPase was drastically reduced in a dose-dependent manner, with complete inhibition at 500 μM prehelminthosporol, while inhibition of ATP hydrolysis was less drastic, with a 35% inhibition at 500 μM toxin. Ca²⁺ uptake was also reduced, although to a lesser extent than proton pumping, with a maximal inhibition of 60% at 500 μM toxin. The 1, 3-β-glucan synthase activity was weakly stimulated at toxin concentrations below 100 μM, with higher concentrations being inhibitory. Taken together, our results indicate that prehelminthosporol exerts its effect in at least two ways. First, it appears to disrupt the membrane barrier and thus seriously interfere with establishment of the proton gradient that drives ion and nutrient uptake and affect the efflux of Ca²⁺ to maintain the low cytoplasmic concentration of Ca²⁺ essential for the function of Ca²⁺ as a messenger in signal transduction. Secondly, it appears to inhibit the enzyme activities. The 1, 3-β-glucan synthase is activated by Ca²⁺ and can operate in the presence of low concentrations of prehelminthosporol (which may induce Ca²⁺ leakage into the cytoplasm and hence activate the enzyme), to produce callose to seal leaky membranes.