Potato Root Exudation and Release of Spongospora subterranea Resting Spore Germination Stimulants are Affected by Plant and Environmental Conditions

Variation in plant and environmental conditions were studied to determine the effect thereof on the exudation of low‐molecular‐weight organic compounds by potato roots. The results of the phytochemical analyses showed that among the conditions investigated, root vigour, potato cultivar, nutrients in incubation solution and temperature influenced the number and the type of primary metabolites released. Moreover, these conditions influenced our detection of compounds known to stimulate germination of resting spores of the pathogen Spongospora subterranea, causal agent of powdery scab and root diseases of potato. We conclude that changes in plant and environmental conditions can affect the release of specific compounds that stimulate germination of S. subterranea resting spores. The impact of the factors affecting potato root exudation on subsequent disease development is discussed.     

Zoosporangial root infection of tomato by Spongospora subterranea in hydroponic and glasshouse culture results in diminished plant growth

Tomato plants are highly susceptible to root infection by Spongospora subterranea and are commonly used as bioassay hosts. The impacts of root infection with S. subterranea on plant productivity and yield have been debated. Recent experiments with potato, the major economic host of S. subterranea, have indicated significantly reduced plant growth and potato yield following heavy infection. However, there have been very few similar studies that have examined the possible impacts of S. subterranea infection on tomato plant growth. Three tomato cultivars, “Grape,” “Roma” and “Truss,” were challenged with S. subterranea inoculum in hydroponic culture. Moderate to severe zoosporangial infections were observed with minor but statistically significant differences in susceptibility among the three tomato cultivars. Zoosporangial root infection in the absence of root gall formation resulted in significantly diminished shoot lengths and plant fresh weights in pathogen challenge tests conducted both in hydroponic culture and glasshouse‐grown plants in potting mix. Root lengths were reduced, but the differences were statistically significant in a single trial only. The findings from this study demonstrate that, as with potato, root infection by S. subterranea can result in reduced tomato plant growth and that root gall production associated with root infection was not necessary for this retardation of growth response. This further suggests that possible yield impacts in other crop species that are hosts for S. subterranea root infection are worthy of examination.     

Arbuscular mycorrhizal fungal propagules in a salt marsh

The tolerance of indigenous arbuscular mycorrhizal fungi (AMF) to stressful soil conditions and the relative contribution of spores of these fungi to plant colonization were examined in a Portuguese salt marsh. Glomus geosporum is dominant in this salt marsh. Using tetrazolium as a vital stain, a high proportion of field-collected spores were found to be metabolically active at all sampling dates. Spore germination tests showed that salt marsh spores were not affected by increasing levels of salinity, in contrast to two non-marsh spore isolates, and had a significantly higher ability to germinate under increased levels of salinity (20) than in the absence of or at low salinity (10). Germination of salt marsh spores was not affected by soil water levels above field capacity, in contrast to one of the two non-marsh spore isolates. For the evaluation of infectivity, a bioassay was established with undisturbed soil cores (containing all types of AM fungal propagules) and soil cores containing only spores as AM fungal propagules. Different types of propagules were able to initiate and to expand the root colonization of a native plant species, but spores were slower than mycelium and/or root fragments in colonizing host roots. The AM fungal adaptation shown by this study may explain the maintenance of AMF in salt marshes.     

Storage of Resting Spores of the Gypsy Moth Fungal Pathogen, Entomophaga maimaiga

The fungal pathogen, Entomophaga maimaiga causes epizootics in populations of the important North American forest defoliator gypsy moth ( Lymantria dispar ). Increasing use of this fungus for biological control is dependent on our ability to produce and manipulate the long-lived overwintering resting spores (azygospores). E. maimaiga resting spores undergo obligate dormancy before germination so we investigated conditions required for survival during dormancy as well as the dynamics of subsequent germination. After formation in the field during summer, resting spores were stored under various moisture levels, temperatures, and with and without soil in the laboratory and field. The following spring, for samples maintained in the field, germination was greatest among resting spores stored in plastic bags containing either moistened paper towels or sterile soil. Resting spores did not require light during storage to subsequently germinate. In the laboratory, only resting spores maintained with either sterile or unsterilized soil at 4°C (but not at 20 or -20°C) germinated the following spring, but at a much lower percentage than most field treatments. To further investigate the effects of relative humidity (RH) during storage, field-collected resting spores were placed at a range of humidities at 4°C. After 9.5 months, resting spore germination was highest at 58% RH and no resting spores stored at 88 or 100% RH germinated. To evaluate the dynamics of infections initiated by resting spores after storage, gypsy moth larvae were exposed to soil containing resting spores that had been collected in the field and stored at 4°C for varying lengths of time. No differences in infection occurred among larvae exposed to fall-collected soil samples stored at 4oC over the winter, versus soil samples collected from the same location the following spring. Springcollected resting spores stored at 4°C did not go into secondary dormancy. At the time that cold storage of soil containing resting spores began in spring, infection among exposed larvae was initiated within a few days after bringing the soil to 15°C. This same pattern was also found for spring-collected resting spore-bearing soil that was assayed after cold storage for 2-7 months. However, after 31-32 months in cold storage, infections started 14-18 days after soil was brought to 15°C, indicating a delay in resting spore activity after prolonged cold storage.     

Production of Vesicular-Arbuscular Mycorrhizal Fungus Inoculum in Aeroponic Culture

Bahia grass (Paspalum notatum) and industrial sweet potato (Ipomoea batatas) colonized by Glomus deserticola, G. etunicatum, and G. intraradices were grown in aeroponic cultures. After 12 to 14 weeks, all roots were colonized by the inoculated vesicular-arbuscular mycorrhizal fungi. Abundant vesicles and arbuscules formed in the roots, and profuse sporulation was detected intra-and extraradically. Within each fungal species, industrial sweet potato contained significantly more roots and spores per plant than bahia grass did, although the percent root colonization was similar for both hosts. Mean percent root colonization and sporulation per centimeter of colonized root generally increased with time, although with some treatments colonization declined by week 14. Spore production ranged from 4 spores per cm of colonized root for G. etunicatum to 51 spores per cm for G. intraradices. Infectivity trials with root inocula resulted in a mean of 38, 45, and 28% of bahia grass roots colonized by G. deserticola, G. etunicatum, and G. intraradices, respectively. The germination rate of G. etunicatum spores produced in soil was significantly higher than that produced in aeroponic cultures (64% versus 46%) after a 2-week incubation at 28°C. However, infectivity studies comparing G. etunicatum spores from soil and aeroponic culture indicated no biological differences between the spore sources. Aeroponically produced G. deserticola and G. etunicatum inocula retained their infectivity after cold storage (4°C) in either sterile water or moist vermiculite for at least 4 and 9 months, respectively.     

Moments of weaknesses – exploiting vulnerabilities between germination and encystment in the Phytomyxea

Attempts at management of diseases caused by protozoan plant parasitic Phytomyxea have often been ineffective. The dormant life stage is characterised by long-lived highly robust resting spores that are largely impervious to chemical treatment and environmental stress. This review explores some life stage weaknesses and highlights possible control measures associated with resting spore germination and zoospore taxis. With phytomyxid pathogens of agricultural importance, zoospore release from resting spores is stimulated by plant root exudates. On germination, the zoospores are attracted to host roots by chemoattractant components of root exudates. Both the relatively metabolically inactive resting spore and motile zoospore need to sense the chemical environment to determine the suitability of these germination stimulants or attractants respectively, before they can initiate an appropriate response. Blocking such sensing could inhibit resting spore germination or zoospore taxis. Conversely, the short life span and the vulnerability of zoospores to the environment require them to infect their host within a few hours after release. Identifying a mechanism or conditions that could synchronise resting spore germination in the absence of host plants could lead to diminished pathogen populations in the field.     

Field and glasshouse experiments on the control of potato mop-top virus

Field observations during 3 yr on a stock of potato cv. Red Craigs Royal partially infected with potato mop-top virus (PMTV) confirmed that the virus was passed by an infected mother plant to only a proportion of its progeny tubers, and showed that in this cultivar symptomless plants gave rise only to symptomless progeny. The elimination of PMTV from stocks can therefore be greatly accelerated by removing symptom-bearing plants. Infected potato tubers were not freed from PMTV by treating them at 37 °C for up to 8 wk. Treating ‘seed’ tubers bearing powdery scabs that contain PMTV-carrying resting spores of Spongospora subterranea with formaldehyde or organo-mercurial fungicide greatly decreased PMTV establishment when the tubers were planted in previously uninfective soil, but fumigation with 2-aminobutane was ineffective. Decreasing the pH of infective soil to 5-0 by applying sulphur greatly decreased the infection of potato cv. Arran Pilot with PMTV and S. subterranea in field experiments, but this treatment did not eliminate either; when the pH of treated soil was raised the transmission of PMTV resumed. Treating infective soil with a range of fungicides greatly decreased the infection of Nicotiana debneyi bait seedlings in glasshouse experiments but only calomel at 75 kg/ha controlled spread of PMTV and 5. subterranea to potato in field experiments. In other field experiments, applying zinc frit, zinc sulphate or zinc oxide to infective soil greatly decreased the spread of both to potato. The amount of zinc required increased with increase in clay content of the soil. However, treatment with zinc compounds did not eliminate PMTV-carrying vectors from soil, and when treated soil was diluted with autoclaved soil many of the bait seedlings planted in the mixture became infected. The zinc frit was phytotoxic because of its boron content but zinc sulphate and zinc oxide caused little or no decrease in tuber yield. The zinc content of potato tubers was increased but not doubled in zinc-treated plots, and during the first year after treatment the zinc content of topsoil decreased greatly. The zinc content of ryegrass grown after potatoes was greater than of potato tubers but did not reach a level considered dangerous to livestock. Treatment of soil with sulphur, zinc oxide or calomel may be useful for small plots used in the early stages of propagation of virus-tested potato clones where there is risk of infection with PMTV.     

The effect of high temperature and fallow period on infection of mung bean and cashew roots by the vesicular-arbuscular mycorrhizal fungus Glomus intraradices

The experiments described were designed to investigate the way in which high temperatures (30°C and above) affected the survival and infectivity of spores of Glomus intraradices formulated as the commercial inoculum NutriLink^TM. Infection of mung bean (Vigna radiata) occurred most rapidly at 30°C compared with either 22° or 38°C, although the final percentage of the root length infected (6 weeks) was similar at all three temperatures. Early rapid infection led to greater plant growth of this species at 30°. In cashew (Anacardium occidentale) no infection occurred at 38°C and this was associated with low plant growth, compared with the other temperatures at which infection reached 40–60% after 4 months. In both species differences in root temperature were associated with marked differences in the morphology and growth of the root systems, with poor root growth at 38°C. Spores of G. intraradices retained infectivity with respect to mung bean for up to 6 weeks in moist fallow soil, although maximum infectivity was observed in soil without a fallow period. The effects of temperature on germination of spores buried in filter paper sandwiches in soil were consistent with the data for infection and growth. Germination was most rapid and reached the highest percentage at 3 weeks at 30°C. Lowest germination was attained at 38°C. We conclude that G. intraradices can retain its infectivity in moist soil at high temperatures, but that the extent to which the plants become infected and hence their response, depends not only on this but also on host factors such as root growth.     

Synchytrium solstitiale: reclassification based on the function and role of resting spores

Studies were made about resting spores of Synchytrium solstitiale, a chytrid that causes false rust disease of yellow starthistle (YST). During evaluation of this fungus for biological control of YST, a protocol for resting spore germination was developed. Details of resting spore germination and study of long-term survival of the fungus were documented. Resting spores from dried leaves germinated after incubating them on water agar at least 7 d at 10-15 C. Resting spores were viable after storage in air-dried leaves more than 2 y at room temperature, suggesting they have a role in off-season and long-term survival of the fungus. Each resting spore produced a single sorus that contained a single sporangium, which on germination released zoospores through a pore. YST inoculated with germinated resting spores developed symptoms typical of false rust disease. All spore forms of S. solstitiale have been found to be functional, and the life cycle of S. solstitiale has been completed under controlled laboratory and greenhouse conditions. Resting spore galls differ from sporangial galls both morphologically and biologically, and in comparison, each sporangial gall cleaves into several sori and each sorus produces 5-25 sporangia that rupture during release of zoospores. For this reason S. solstitiale should be reclassified as diheterogallic sensu Karling (Am J Bot 42:540-545). Because resting spores function as prosori and produce an external sorus, S. solstitiale is best placed in into the subgenus Exosynchytrium.     

Plasmodiophora brassicae: a review of an emerging pathogen of the Canadian canola (Brassica napus) crop

Plasmodiophora brassicae causes clubroot disease in cruciferous plants, and is an emerging threat to Canadian canola (Brassica napus) production. This review focuses on recent studies into the pathogenic diversity of P. brassicae populations, mechanisms of pathogenesis and resistance, and the development of diagnostic tests for pathogen detection and quantification. TAXONOMY: Plasmodiophora brassicae is a soil-borne, obligate parasite within the class Phytomyxea (plasmodiophorids) of the protist supergroup Rhizaria. DISEASE SYMPTOMS: Clubroot development is characterized by the formation of club-shaped galls on the roots of affected plants. Above-ground symptoms include wilting, stunting, yellowing and premature senescence. DISEASE CYCLE: Plasmodiophora brassicae first infects the root hairs, producing motile zoospores that invade the cortical tissue. Secondary plasmodia form within the root cortex and, by triggering the expression of genes involved in the production of auxins, cytokinins and other plant growth regulators, divert a substantial proportion of plant resources into hypertrophic growth of the root tissues, resulting in the formation of galls. The secondary plasmodia are cleaved into millions of resting spores and the root galls quickly disintegrate, releasing long-lived resting spores into the soil. A serine protease, PRO1, has been shown to trigger resting spore germination. PHYSIOLOGICAL SPECIALIZATION: Physiological specialization occurs in populations of P. brassicae, and various host differential sets, consisting of different collections of Brassica genotypes, are used to distinguish among pathotypes of the parasite. DETECTION AND QUANTIFICATION: As P. brassicae cannot be cultured, bioassays with bait plants were traditionally used to detect the pathogen in the soil. More recent innovations for the detection and quantification of P. brassicae include the use of antibodies, quantitative polymerase chain reaction (qPCR) and qPCR in conjunction with signature fatty acid analysis, all of which are more sensitive than bioassays. RESISTANCE IN CANOLA: Clubroot-resistant canola hybrids, recently introduced into the Canadian market, represent an important new tool for clubroot management in this crop. Genetic resistance must be carefully managed, however, as it has been quickly overcome in other regions. At least three resistance genes and one or two quantitative trait loci are involved in conferring resistance to P. brassicae. Root hair infection still occurs in resistant cultivars, but secondary plasmodia often remain immature and unable to produce resting spores. Fewer cell wall breakages occur in resistant hosts, and spread of the plasmodium through cortical tissue is restricted. More information on the genetics of clubroot resistance in canola is needed to ensure more effective resistance stewardship. USEFUL WEBSITES: http://www.canolacouncil.org/clubroot/resources.aspx, http://tu-dresden.de/die_tu_dresden/fakultaeten/fakultaet_mathematik_und_naturwissenschaften/fachrichtung_biologie/botanik/pflanzenphysiologie/clubroot, http://www.ohio.edu/people/braselto/plasmos/     

Breaking dormancy in spores of the arbuscular mycorrhizal fungus Glomus intraradices: a critical cold-storage period

To elucidate the effect of cold storage on spore dormancy in the arbuscular mycorrhizal (AM) fungus Glomus intraradices, spores were cold stratified at 4 degrees C, for either 0, 3, 7, 14, 90 or 120 days, prior to germination tests at 25 degrees C. The results showed that cold stratification longer than 14 days significantly increased spore germination. Moreover, the longer cold storage periods clearly reduced spore mortality from 90% to 50% and considerably altered the hyphal growth pattern. Long polarized hyphae were only observed after cold stratification periods longer than 14 days, involving consequences for root infectivity. The results clearly show that environmental factors, e.g., coldness, can affect the physiology of AM fungal spores.     

Susceptibility of opium poppy and pyrethrum to root infection by Spongospora subterranea

Spongospora subterranea, which causes powdery scab of potato, infects a diverse range of plant species. Crop rotation as a powdery scab management tool will be compromised if pathogen hosts exist between potato crops. Opium poppy (Papaver somniferum) and pyrethrum (Tanacetum cinerariifolium) are important crops within intensive vegetable production rotations in NW Tasmania. Measurements of S. subterranea soil inoculum within a commercial field showed pathogen amounts were substantially elevated following an opium poppy crop, which suggested host status. In glasshouse testing, opium poppy and pyrethrum were confirmed as hosts of S. subterranea, with opium poppy the more susceptible of the two. Both species were less susceptible than tomato, a known host. Observations of early growth suggested inoculation impacts on all three plant species, although at 16 (tomato and opium poppy) or 26 (pyrethrum) weeks postinoculation, only tomato had significantly reduced shoot and root development. The role of rotation crops in inoculum persistence and the possible role of S. subterranea as a minor pathogen of nonpotato crops are discussed.     

Honey-coloured,sessile Endogone spores: II. Changes in fine structure during spore development

Summary The fine structure of honey-coloured, sessile Endogone spores is described from initiation of the mother spore to dormancy of the resting spore. Three unusual organelles occur viz. pigment granules, large crystals and selfduplicating bacteria-like organisms. The first two are very numerous, and are specifically associated with spore formation. The pigment granules are involved in the deposition of the honey-coloured wall, and change into myelin-like figures when cytoplasm moves from the mother into the resting spore. The crystals, whose function is not known, are most conspicuous just before the resting spore reaches dormancy. The bacteria-like organisms, which may be actinomycete spores living symbiotically in the fungus, multiphy greatly as the spore enters dormancy. The dormant spore contains very little cytoplasm compressed into a fine network between very large polygonal oil globules and large round bodies thought to contain a storage polysaccharide.     

The effectivity of arbuscular mycorrhizal fungi from high input conventional and organic grassland and grass-arable rotations

The effectivity of arbuscular mycorrhizal spores in promoting growth of Allium ameloprasum L. cv. Musselburgh and Trifolium repens L. cv. Menna was tested for inocula from three soil series under long term organic or intensive, conventional grass and grass-arable rotations. For two soil series, Allium responses to inocula from soils recently converted to organic fanning were also assessed. Finally, Trifolium root fragments were used to inoculate Allium so as to evaluate responses to this inoculation procedure. Plants were sown into previously sterilised, matched soils from organic farms with no nutrient input. Mycorrhizal treatments generally increased growth for Allium. However, for Trifolium, infection decreased growth in the most fertile soil and gave an increase only in the least fertile. In the least fertile soil, inocula from organic farms were more effective than those from conventional farms. For Trifolium (all soils) and for Allium (least fertile soil), there was evidence of more efficient uptake of phosphorus in plants inoculated with spores from organic farms. The pattern of Allium response to inoculation with spores from conventional, conversion and organic sources was not consistent between soil type, but there was evidence of lower root infection for conversion compared with organic inocula and of a trend towards higher infectivity as the time period under organic management increased. Inoculating Allium with AMF root fragments produced a plant response similar to that obtained when spores were used, confirming that spore viability was not the sole factor influencing AMF effectivity in earlier experiments. Intensive farming practices may reduce the effectiveness of indigenous arbuscular mycorrhizal populations, particularly where fertiliser inputs are high and inherent fertility is low. This could have practical implications where high input farms are converted to organic management.     

Annual dormancy cycles in buried seeds of shrub species: germination ecology of Sideritis serrata (Labiatae)

The germination ecology of Sideritis serrata was investigated in order to improve ex‐situ propagation techniques and management of their habitat. Specifically, we analysed: (i) influence of temperature, light conditions and seed age on germination patterns; (ii) phenology of germination; (iii) germinative response of buried seeds to seasonal temperature changes; (iv) temperature requirements for induction and breaking of secondary dormancy; (v) ability to form persistent soil seed banks; and (vi) seed bank dynamics. Freshly matured seeds showed conditional physiological dormancy, germinating at low and cool temperatures but not at high ones (28/14 and 32/18 °C). Germination ability increased with time of dry storage, suggesting the existence of non‐deep physiological dormancy. Under unheated shade‐house conditions, germination was concentrated in the first autumn. S. serrata seeds buried and exposed to natural seasonal temperature variations in the shade‐house, exhibited an annual conditional dormancy/non‐dormancy cycle, coming out of conditional dormancy in summer and re‐entering it in winter. Non‐dormant seeds were clearly induced into dormancy when stratified at 5 or 15/4 °C for 8 weeks. Dormant seeds, stratified at 28/14 or 32/18 °C for 16 weeks, became non‐dormant if they were subsequently incubated over a temperature range from 15/4 to 32/18 °C. S. serrata is able to form small persistent soil seed banks. The maximum seed life span in the soil was 4 years, decreasing with burial depth. This is the second report of an annual conditional dormancy/non‐dormancy cycle in seeds of shrub species.     

Fire and flood: Soil‐stored seed bank and germination ecology in the endangered Carrington Falls Grevillea (Grevillea rivularis,Proteaceae)

Abstract Seed germination is dependent on the interaction between the dormancy state of a seed and the presence of favourable environmental conditions. Thus, the spectacular pulse of seedling recruitment in many Australian vegetation communities following disturbances such as fire can be attributed to changes in microsite conditions and/or the dormancy‐breaking effect of the disturbance on accumulated seed banks. Grevillea rivularis is a threatened species endemic to the area immediately above Carrington Falls in the NSW Southern Highlands. Most of the population is confined to the riparian vegetation zone in woodland and heath, and is therefore subject to periodic disturbance from fire and flood. For this species, a pulse of seedling recruitment has been recorded after fire, flood and mechanical soil disturbance. The aims of this study were to examine the density and vertical distribution of the soil‐stored seed bank and to investigate the role of heat and scarification as cues for germination of fresh and soil‐stored seed. There was a large seed bank under the canopies of established individuals (194 ± 73 seeds m^?2) and most seeds were found in the 0–2 cm and leaf‐litter layers of the soil profile. The germination response of soil‐stored and fresh seed was examined using a hierarchical series of laboratory experiments. Seeds of G. rivularis showed marked dormancy polymorphism. Thirty‐six percent of soil‐stored seed germinated without treatment, whereas no untreated fresh seeds germinated. Scarification or heating caused significant germination of dormant soil‐stored seed, but only scarification resulted in germination of dormant fresh seeds. These results highlight important differences in the dormancy state of soil‐stored and fresh seed. Thus, being a riparian species in a fire‐prone environment, the dormancy mechanisms in seeds of G. rivularis suit this species to disturbance by both fire and flood.     

Effect of trehalose on the viability of sporangiospores of the mucorous fungus <Emphasis Type="Italic">Blakeslea trispora</Emphasis>

Sporangiospores of Blakeslea trispora are in a state of exogenous dormancy, and water is the key factor controlling their germination. A wide range of carbohydrates, ammonium salts, and yeast extract had a weak stimulating effect (less than 50%) on spore germination, whereas amino acids could significantly inhibit this process. Cultivation of B. trispora on sporogenous sucrose- and trehalose-containing media (S and T spores, respectively) resulted in significant changes in spore formation, as well as in the chemical composition of spores and their viability. In the presence of trehalose, the amount of spores increased twofold; spore viability during storage increased as well. All changes in the carbohydrate composition of the cytosol and in the composition of the spore membrane lipids indicated that the dormancy of T spores was deeper than that of S spores, which has a favorable effect on their viability.     

“INTERNAL THECAE” OF EUNOTIA SOLEIROLII (BACILLARIOPHYCEAE): DEVELOPMENT,STRUCTURE AND FUNCTION AS RESTING SPORES1

The “double thecae” or “internal septa” of Eunotia soleirolii (Kütz.) Rabenh, are shown to represent the thecae of resting spores, as characterized by their physiology, as well as morphology. They differ from all resting spores of centric diatoms by the formation of both their valves as a result of unequal cell divisions; and, from the majority of centric spores by the presence of several girdle bands in both their thecae. Spore formation can be induced by high or low pH, high temperature (24 C), and iron, silica, phosphate or nitrate deficiencies, whereas low temperatures defer it. Spores do not germinate directly, but dormancy can be removed by dark treatments (–2 to 15 C) for a minimum of 4–5 wk. Longer dark treatments result in higher germination rates. At 15 C, a minimum of 2 mo is required and 4 mo is better. Heat treatments (27–42 C) are ineffective, but may shorten the dormancy-breaking subsequent cold period. Instances of secondary dormancy, as well as relative dormancy, were observed. Germination usually occurs in the light between 2 and 21 C. An equal division of the spore is followed by unequal divisions of both new cells with only the two resulting large cells being viable. The experiences in the laboratory aided the discovery of stages of spore germination in nature.     

Alleviation of salt stress by plant growth regulators and IAA producing bacteria in wheat

The action of phytohormone producing bacteria and plant growth regulators on germination and seedling growth of wheat under saline conditions were studied. Seed dormancy enforced by salinity (100 mM NaCl) was substantially alleviated and the germination was promoted by gibberellin, auxin, zeatin, and ethephon from 54 to 97%. The IAA producing bacterial strains Pseudomonas aureantiaca TSAU22, Pseudomonas extremorientalis TSAU6 and Pseudomonas extremorientalis TSAU20 significantly increased seedling root growth up to 25% in non-salinated conditions and up to 52% at 100 mM NaCl, compared to control plants. It is concluded that growth regulators considerably alleviated salinity-induced dormancy of wheat seeds. The facts mentioned above make it possible to recommend root colonizing bacteria that produce phytohormone to alleviate salt stress of wheat grown under conditions of soil salinity.     

Evaluation of Clove Essential Oil as a Mycobiocide Against Rhizopus stolonifer and Fusarium solani,Tuber Rot Causing Fungi in Yam (Dioscorea rotundata Poir.)

To exploit natural products for plant disease control, the essential oil of Syzygium aromaticum (L.) Merr. & Perr. (clove) was investigated for its antifungal activity against Rhizopus stolonifer and Fusarium solani, the postharvested yam tuber rot pathogens. The essential oil was obtained by hydrodistillation using a Clevenger‐type apparatus. The chemical composition of the oil was determined by gas chromatography and gas chromatography coupled with mass spectrometry. Antifungal activities of the oil were tested in vitro against mycelia growth and spores germination. In situ tests were conducted on healthy yam tubers, and necrosis symptoms were assessed. Results showed that eugenol (79.4%), eugenylacetate (9.2%) and isocaryophyllene (7.0%) were the major components. The oil exerted antifungal activities with total inhibition (TI) of the mycelia growth of R. stolonifer and F. solani was recorded at 200 and 300 ppm, respectively, while TI of spores germination was recorded at 31.25 and 250 ppm, respectively. For the standard fungicide (Ridomil^®), TI value of mycelia growth was 1600 ppm for the both pathogens, while TI of spores germination were 200 ppm and 1600 ppm, respectively, for Rhizopus and Fusarium. In situ tests showed complete inhibition of yam tuber rot when the essential oil was applied at 2000 ppm for preventive tests. This oil also reduced significantly (P ≤ 0.05) necrosis development on yam tuber for curative test at the same concentration. Total inhibition of the necrosis by Ridomil (3000 ppm) was observed only for Rhizopus on preventive test. There were positive correlations between the oil concentration and the reduction of necrosis cause by R. stolonifer and F. solani. These findings showed that clove essential oil may serve as environmental friendly bio‐fungicide for the management of postharvest yam tuber rot.