Witches' broom disease on cocoa in Rondonia, Brazil: basidiocarp production on detached brooms in the field

Fruiting of Crinipellis perniciosa was assessed in relation to various climatic factors using dead witches' brooms detached from cocoa, either suspended in the canopy or laid on leaf litter under cocoa. Induction of basidiocarp production occurred over approximately 60 days with rain. Basidiocarps were finally observed on about 85% of brooms in all samples examined and induction of fruiting required at least 17 rainy days, although brooms with a minimum age of 2 years produced basidiocarps about 25 days before brooms with a minimum age of 1 year. Groups of brooms showed distinct cycles of basidiocarp production, and up to 50% of fruiting occurred on dead, attached leaves, Fruiting was reduced on brooms on leaf litter and basidiocarps survived for less time than when brooms were suspended in canopy. Brooms were most productive with moderate amounts of wetness per day, and less than 4 h or more than 20h was inhibitory. No fruiting occurred at mean air temperatures higher than 30°C or lower than 20°C. The microclimate of the litter layer is discussed in relation to reduction of fruiting caused by pruning of brooms.     

Analysis of resistance to witches’ broom disease (Moniliophthora perniciosa) in flower cushions of Theobroma cacao in a segregating population

This study evaluates resistance to witches’ broom disease in flower cushions of Theobroma cacao under field conditions. The aim was to determine optimal inoculation methods to evaluate the disease incidence using flower cushions in the field. A segregating mapping population of 580 trees (cultivar TSH 1188 × CCN 51) was analysed under two field conditions: high and low inoculum levels (in different years), corresponding respectively to trees with or without dried witches’ brooms hanging on the trees and producing basidiocarps. The number of newly formed cushion brooms in each tree was counted by the conventional method, and also the healthy and infected flower cushions in three 30 cm‐long regions along the trunk and the two main branches. The field inoculation methods discriminated between genotypes, with a 26% increase in disease incidence by Moniliophthora perniciosa at high inoculum. Two different segregation patterns were also observed: 27:27:9:1 under low, and 27:9:9:9:3:3:3:1 under high inoculum potential. It was also determined that at least 20 flower cushions were needed to accurately determine the percentage of infection. These methodologies allowed identification of the extreme phenotypes in this mapping population, and can therefore facilitate the detection of sources of resistance to witches’ broom disease.     

Assessing resistance to Crinipellis perniciosa using cocoa callus

Callus cultures from cocoa clones reported to have different susceptibilities to Brazilian isolates of Crinipellis perniciosa , causal fungus of witches' broom disease, including Catongo (susceptible). Scavina-6, CAB 64 and CAB 67 (resistant) were inoculated with basidiospores of C. perniciosa and the subsequent development of mycelia was assessed. Generally, on Catongo callus, mycelium similar to that found in green brooms (parasitic type), developed abundantly but on callus from Scavina-6 and the two CAB clones, mycelial growth was relatively poor and tended to be similar to that in dry brooms and on agar media (saprotrophic type). There was often considerable variation in fungal development on individual callus cultures from the same clone, but there was an overall consistency between successive experiments in ranking the callus material in terms of fungal development. However, growth of isolates from Colombia and Trinidad was similar on callus from Scavina-6 which in the field is known to reac differentially to these two isolates. Experiments with isolates from Colombia indicated that inoculum from basidiocarps produced on brooms which were kept for long periods in cabinets to induce fruiting had a decreased ability to colonize callus.     

Production and Germination of Conidia of Trichoderma stromaticum, a Mycoparasite of Crinipellis perniciosa on Cacao

ABSTRACT Growth characteristics of the fungus Trichoderma stromaticum, a mycoparasite on the mycelium and fruiting bodies of Crinipellis perniciosa, the causal agent of witches'-broom disease of cacao, were evaluated under controlled environmental conditions. The ability of T. stromaticum to produce conidia and germinate on dry brooms was evaluated at three constant temperatures (20, 25, and 30 degrees C) and two constant relative humidities (75 and 100%). T. stromaticum produced abundant conidia on brooms at 100% relative humidity and incubation temperatures of 20 and 25 degrees C, but none at 30 degrees C. Sporulation of T. stromaticum was not observed at 75% relative humidity at any temperature. At 100% relative humidity and either at 20 or 25 degrees C, treatment of brooms with T. stromaticum suppressed C. perniciosa within 7 days. In contrast, at 30 degrees C, treatment with T. stromaticum had no effect on the pathogen in brooms maintained at either 75 or 100% relative humidity. Mycelium of C. perniciosa grew from brooms at all temperatures at 100% relative humidity. Conidial germination on broom tissue approximated 80% at temperatures from 20 to 30 degrees C. Results suggest that applying T. stromaticum under high-moisture conditions when the air temperature is below 30 degrees C may enhance the establishment of this mycoparasite in cacao plantations.     

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.     

Witches' broom disease on cocoa in Rondonia, Brazil: pod infection in relation to pod susceptibility, wetness, inoculum, and phytosanitation

Infection of cocoa pods by Crinipellis perniciosa was examined in the field in relation to pod susceptibility and variations in inoculum and climate over 2 years. The disease was incubated for 13 or 14 weeks in pods inoculated when 2 and 4 weeks old, and for 12 weeks in pods inoculated 6-12 weeks old. Pods were uniformly susceptible to natural infection during the 12 weeks after pollination. Over 75% of rain showers started between 12.00 and 02.00 h, providing sufficient water to rehydrate basidiocarps as well as to wet pods. Pods remained wet after rainfall for decreasing periods when rain occurred after 13.00 h. Incidence of pod disease was shown to be related to the number of wetness periods longer than 4 h, and to the number of open turgid basidiocarps occurring 12 weeks earlier. Wetness periods had a greater influence on disease. Pod infection was modelled by assuming that infection was limited by either wetness periods or turgid basidiocarp numbers; critical numbers of wetness periods when both factors were of equal weight were defined by an exponential equation. The model was used to predict reductions in pod disease associated with different amounts of broom removal in each year.     

Genetic and Biological Diversity of Trichoderma stromaticum, a Mycoparasite of the Cacao Witches'-Broom Pathogen

ABSTRACT The witches'-broom disease, caused by the basidiomycete Crinipellis perniciosa, is the most limiting factor for cacao cultivation in Brazil. Trichoderma stromaticum is a mycoparasite of the witches'-broom pathogen of cacao that is currently being applied in the field to manage the disease in Bahia State, Brazil. In this work, molecular and traditional methods were used to study the genetic and biological diversity of this mycoparasite. Ninety-one isolates, mostly collected from farms not sprayed with the fungus, were analyzed by amplified fragment length polymorphisms (AFLP), which showed that two genetic groups (I and II) of T. stromaticum occur in Bahia State. This classification of T. stromaticum into two distinct AFLP groups was also in agreement with several other characteristics, including growth on agar media at different temperatures and sporulation on infected stem segments (broom pieces) and rice grains. Group II favors higher temperatures compared with group I. The genetic and biological differences of the isolates, however, were not evident in field experiments, where sporulation was evaluated on the surface of brooms under natural conditions. Our results show that there is considerable genetic and biological diversity within T. stromaticum in Bahia and other cacao-growing regions of South America that are affected by the witches'-broom disease. This diversity could be explored in the development of efficient biological control agents against the disease. Factors that may affect the application and performance of this biocontrol agent in the field, such as sporulation on rice substrate and on the brooms and growth at various temperatures, are discussed.     

Of mushrooms and chocolate trees: aetiology and phylogeny of witches' broom and frosty pod diseases of cacao

The troubled history of the two major diseases of the chocolate tree (Theobroma cacao) in South America, witches' broom and frosty pod, is reviewed, concentrating on critical aspects of the aetiology as well as the phylogeny of the causal agents. Both diseases are caused by sister species within the genus Moniliophthora, belonging to the Marasmiaceae family of mushrooms. The witches' broom pathogen, Moniliophthora perniciosa, evolved on the Amazonian side of the Andes and induces brooms not only in cacao and its relatives in the genera Theobroma and Herrania (Malvaceae), but also in species in the plant families Bignoniaceae, Malpighiaceae and Solanaceae, on which the mushrooms (basidiomata) are produced. Moniliophthora roreri, the type species of the genus, evolved as a pod pathogen on endemic Theobroma species on the western side of the northern Andean Cordillera. Because Moniliophthora was described originally as the asexual form of an unknown basidiomycete, the generic diagnosis is amended here to accommodate species with agaricoid basidiomata. In addition, the new variety M. roreri var. gileri is designated for the morphotype occurring on Theobroma gileri, in northwest Ecuador. Cytology studies indicate that the supposed conidia of M. roreri are, in fact, sexual spores (meiospores) and it is posited that the fruiting structure represents a much‐modified mushroom. Finally, based on preliminary data from pathogenicity testing, it is hypothesized that the true causal agent of both diseases is an as yet unidentified infectious agent vectored into the host by the fungus.     

Changes in morphology and measurement of cytokinin levels during the development of witches' brooms on cocoa

The anatomy of diseased stems from brooms induced on cocoa ( Theobroma cacao ) by the fungus Crinipellis perniciosa was examined using the scanning electron microscope, and the amounts of selected cytokinins were measured by immunoassay.
Although the diseased stems were wider in diameter with a lower dry weight:fresh weight ratio than stems of healthy plants, the overall internal organization of tissues remained unchanged, and all the cell types were present. However, the vascular tissue was less differentiated in diseased stems since xylem vessels were absent and the ratio of phloem fibres and sieve tubes to phloem parenchyma was reduced. Cell sizes in diseased stems were larger for all tissues but cell numbers were unchanged except for xylem where fewer cells were seen.
Cytokinins (zeatin, zeatin riboside, isopentenyl adenine and isopentenyl adenosine) were measured in healthy plants and diseased stems at specific stages of disease development. Of these cytokinins only zeatin riboside was present in significantly greater amounts in diseased tissue. The likely role of plant growth regulator balance in stem enlargement and proliferation in witches' broom disease of cocoa is discussed.     

Pathogenic variability amongst isolates of Crinipellis perniciosa from cocoa (Theobroma cacao)

Dry witches' brooms from cocoa were imported from various areas within Bolivia, Brazil, Colombia, Ecuador, Trinidad and Venezuela. Basidiocarps of Crinipellis perniciosa were induced to form on these brooms and seedlings of different types of cocoa were inoculated with basidiospores either on the hypocotyl or cotyledon bud. Host reactions were assessed mainly by recording stem base swelling and broom development at the cotyledon node (hypocotyl inoculations) or the extent of swelling and branching of shoots (cotyledon bud inoculations).
Results from 30 experiments indicated considerable diversity amongst isolates in inducing disease symptoms, but suggested that two groups or populations of C. perniciosa exist on cultivated cocoa. One group (A), comprising isolates from Bolivia and Pichilingue (Ecuador) and most isolates tested from Colombia, induced severe symptoms on cocoa with Scavina 6 as one parent; the other group (B), comprising isolates from Brazil, Trinidad and Venezuela, did not. Within these groupings variants could be further distinguished by particular host reactions. Isolates from Ecuador, especially from the Oriente, a centre of diversity for Theobroma cacao, showed a range of pathogenicity comparable to that found amongst isolates from cultivated cocoa over a much wider area.     

Relationships Between Black Pod and Witches'-Broom Diseases in Theobroma cacao

ABSTRACT Field observations were conducted from 1998 to 2001 at the International Cocoa Genebank, Trinidad, to evaluate 57 cacao clones for resistance to black pod (BP) and witches'-broom (WB) diseases (caused by Phytophthora sp. and Crinipellis perniciosa, respectively). Each month ripe pods were harvested and the number of healthy and diseased was recorded. The number of brooms on vegetative shoots was recorded three times a year on selected branches. Twenty-three clones showed less than 10% of infection for both BP and WB on pods. Among those, eight clones showed an absence of brooms on the observed branches: IMC 6, MAN 15/60 [BRA], PA 67 [PER], PA 195 [PER], PA 218 [PER], PA 296 [PER], PA 303 [PER], and POUND 32/A [POU]. Broad-sense heritability was estimated at 0.38 and 0.57 for WB disease on pods and shoots, respectively, and at 0.51 for BP disease. Genetic correlation between WB disease on pods and on shoots was low and estimated at 0.39, whereas the correlation between WB and BP diseases on pods was 0.48. To choose putative parents for breeding schemes, it is suggested that clones are first assessed for their level of resistance to WB on shoots, and the most promising individuals are screened for BP with a detached pods test. Further studies are needed to confirm whether the level of resistance to WB on pods can be predicted using an early test on seedlings.     

<Emphasis Type="Italic">Helicobasidium mompa</Emphasis> isolates from sweet potato in continuous monoculture fields

 Isolates of the violet root rot fungus Helicobasidium mompa were collected from herbaceous and tree plants. Their host preference was studied by inoculation experiments using carrots, sweet potatoes, and apple stocks. It was found that sweet potato isolates from Kyushu produced infection cushions on carrots and sweet potatoes but not on apple stocks. Other isolates did not show host preference. Sweet potato isolates were also characterized by ready hyphal mass (sclerotium) production. They were thought to have adapted to the habitat with high disturbance by annual tillage. Received: July 15, 2002 / Accepted: September 26, 2002     

Summer-grown seed potatoes: an attempt to produce seed for the spring season in Israel

Seed potatoes were grown in Israel during summer (June-October) at Ramat haNegev (the Negev plateau). This is the hottest season in the semi-arid climate prevailing in the region. Because of the high temperature, no tuberization occurred in cv. ‘Up-to-Date; and ‘Draga’ and ‘Cara’ showed a marked yield reduction. Reasonable yields were, however, obtained at Ramat haNegev during summer growth with the cvs. ‘Désirée’, ‘Blanka’, ‘Spunta’ and ‘Alpha’. At Bet Dagan (coastal plain) Désirée grown during summer met with many problems and yields were negligible, probably because of the higher night temperature as compared with Ramat haNegev. Growth during summer in Ramat haNegev offers the advantages of relative isolation from other potato fields (potatoes are not grown in summer) and low aphid activity, in contrast to the common growing seasons — spring and autumn. Insecticide application and planting high quality seeds — imported elite or Class A — in summer, reduced potato leaf roll virus (PLRV) infection to an average of 4.5% in 1984 and 1.8% in 1985, in comparison with an average of 20.8% in 1979, when poor quality seeds were used and no insecticides were applied. Infection with potato virus Y (PVY) was 1.7% in 1984 and 0.1% in 1985. The yielding capacity in spring 1980 of seed potato produced at Ramat haNegev in the summer of 1979, was significantly lower than that of imported and Golan Heights seed. This is attributed mainly to the high level of virus infection of the former seeds. PLRV-free seed potatoes from the same batch yielded similarly to imported seed potatoes or seed potatoes from the Golan Heights. This was reconfirmed in the 1985 experiment in which Ramat haNegev seeds were much less infected with PLRV and produced yields similar to the imported and Golan ones. Seed potatoes from Ramat haNegev behave like young seeds when planted in the spring. Therefore, their yielding capacity probably increases the earlier the harvest and the longer the subsequent growing period.     

凹缘菱纹叶蝉的越冬习性与传播枣疯病的关系

 发现凹缘菱纹叶蝉(Hishimonus seⅡatus Uhler)是传播枣疯病的主要媒介叶蝉之一.在我国华北,它于8月下旬开始迁往枣林附近的松(Pinus tabulaeformis Carr)柏(Platycladus orientalis Franco)上,以成虫越冬,但不在松、柏树上繁殖世代.9月中旬迁移越冬进入盛期,10月份松、柏树上的虫口已达高峰;翌年4月中旬转移活动,5月上旬枣芽萌发期间全部迁离松,柏树.经接种实生枣苗证明:越冬后成虫仍保持较高的传染枣疯病的能力,每苗接虫2—5头,枣苗发病率达7.1—76:2%.试验也证明,带毒叶蝉的其子代叶蝉不再带毒。本试验同时证明,带病叶蝉不单纯是传病媒介,而且是一种相当持久的枣疯病侵染源,防治上应予以充分注意.     

不同时期水分胁迫对水稻生长特性和产量形成的影响

通过网室盆栽试验,研究了不同时期水分胁迫对杂交稻生长和产量的影响。结果表明,中优6号各生育期不同程度水分胁迫都会影响水稻产量,减产最大的时期是穗分化期和开花期,产量比对照下降49.4%和30.7%。水稻分蘖期受水分胁迫对穗数影响最大。穗分化期水分胁迫对总粒数和结实率影响最大,开花期水分胁迫对千粒重影响最大,结实率与穗抽出率存在正相关,相关系数达到0.9211。穗分化期和开花期重度水分胁迫,穗抽出率明显下降。在水分胁迫条件下,光合速率下降主要发生在下午。讨论了水分胁迫对水稻产量和生长的影响。     

Solarization; a physical control method for weeds and parasitic plants (Orobanche spp.) in Mediterranean agriculture

Summary. Experiments were conducted between 1985 and 1988 to evaluate the effect of duration and time of solarization on the control of two broom rapes ( Orobanche aegyptiaca Pers., O. crenata Forsk.) and other weeds common in the fields of faba bean ( Vicia faba L.) and lentil ( Lens culinaris Medik.) in West Asia. Solarization was performed by applying clear polyethylene sheet to the soil for 0, 10, 20, or 40 days in 1985/86, 0, 30, or 40 days in 1986/87 and 0, 20, 30, 40 or 50 days in 1987/88 during the hot season (July and August) and for 50 days immediately before sowing during milder weather (September and October) in 1986/87. Best control was obtained with solarization for 30–50 days in the hot season. Maximum soil temperature under polyethylene at 5 cm was 55°C in 1985, 48°C in 1986 and 57°C in 1987. Broomrape dry weight decreased in the first experiment by more than 90% in both faba bean and lentil fields. Solarization controlled broom-rape and other weeds in the following two seasons when the soil was left undisturbed during the season of the treatment.     

In vitro effects of temperature and wet periods on infection of oilseed rape by Alternaria brassicae

The influence of temperature and duration of wet periods on infection of oilseed rape by Alternaria brassicae was studied on detached leaves and pods, leaf disks and intact seedlings. Infections increased with age of leaf and the interaction between temperature and leaf age was highly significant. On older leaves infection was optimal at 25°C. There were many infections also at 15, 20 and 29 C but relatively few infections at 10°C. On pods most infections were observed at 20 C, the highest temperature studied. Infection at each temperature increased progressively with duration of surface wetness. The minimum wet periods for infection of leaves were 3 h at 20–25°C, 4 h at 15°C, 6–9 h at 10 C and 12–24 h at 5 C and for infection of pods, between 6 h and 9 h at 10°C and 6 h (or less) at 15°C and 20 C. On leaves, dry periods interrupting wet periods limited lesion development to that obtained with the initial wet period only; on pods some further infections developed when pods were re-wetted. Dry periods of 3 h and 6 h following the inoculation of pods reduced subsequent infection but there was no further reduction by longer periods of drying to 48 h.     

Factors affecting infection cushion development by Rhizoctonia solani AG-1 IA and IB on soybean leaves

Infection cushions were formed by isolates of Rhizoctonia solani , anastomosis group 1 IA (AG-1 IA, aerial blight) and AG-1 IB (web blight) on leaves of all 10 soybean cultivars tested. Isolates of AG-1 IA and IB did not form infection cushions on soybean leaf surface replicas of either resistant or susceptible cultivars. More infection cushions were formed by isolates of AG-1 IA and IB on collodion membranes placed over leaves of susceptible cultivars compared with resistant cultivars. Isolates of AG-1 IC. AG-4 and AG-5, also formed infection cushions on soybean leaves. However, the isolates of other anastomosis groups did not form infection cushions on soybean leaves. Differential induction of infection cushion formation by the leaves of various plant species was observed, AG-1 IA formed infection cushions on more graminaceous hosts than AG-1 IB, Our results suggest that a chemical stimulus is needed for infection cushion formation. Glucose and 3- O -methylglucose repressed disease severity caused by AG-1 IA and IB isolates to the same extent. Disease severity and the number of infection cushions formed on leaves of ten soybean cultivars were correlated. Fewer infection cushions were formed on resistant cultivars than on susceptible cultivars.     

An optimized screening method for identifying levels of resistance to Crinipellis perniciosa in cocoa (Theobroma cacao)

The effects of host age, leaf number, host type (clone or seedling), pathogen spore concentration and incubation time on inoculation with Crinipellis perniciosa (witches' broom disease of cocoa) were studied in greenhouse experiments using susceptible cocoa genotypes. Three methods of inoculation (agar-drop, water-drop and spray) were also tested. An optimized inoculation method was selected and tested for its repeatability as well as its ability to discriminate between various levels of resistance to C. perniciosa in cocoa. The optimized method (350 000 viable basidiospores per mL, 60 h incubation, agar-drop technique) produced 100% infection repeatedly, on both clonal and seedling plants of a susceptible genotype. Seedling age (2–12 months) and leaf number did not significantly affect the percentage of plants with symptoms or broom characteristics. This method discriminated effectively between the various levels of resistance in 14 cocoa genotypes and is recommended as an inoculation method to identify levels of resistance in germplasm collections. Symptom severity was shown to be a better measure of resistance than infection success.     

近20年来祁连山区植被变化特征分析

利用1982-2003年8 km分辨率的 NDVI数据集,分析了祁连山区植被覆盖的年、年际、年代变化特征.结果表明:祁连山区植被覆盖总体上自东向西递减,其年变化NDVIm都为单峰型;祁连山区各段植被覆盖的年际NDVIy、年代变化NDVIn趋势有所不同:祁连山西中段植被的NDVIy 变化趋势基本相似,20世纪80年代变化较平稳,90年代变化幅度最大,1995年以后植被明显开始增加,祁连山东段植被的 NDVIy 从1982年开始呈增加趋势,直到1997年以后下降;祁连山西中段1982-1985,1986-1990,1991-1995,1996-2000,2001-2003年5个阶段植被的 NDVIn最大值出现在2001-2003年,最小值出现在1991-1995年,而祁连山东段最大值出现在1996-2000年,最小值出现在1982-1985年;祁连山各段5～9月植物生长期植被的NDVIpy,NDVIpn变化与年的NDVIy,NDVIn变化相似.