Implications of bacterial contaminated seed lots and endophytic colonization by Pseudomonas fuscovaginae on rice establishment

The economic impact of seedborne bacterial diseases on rice production provides a major motivation for research on seed health. This paper reports on the endophytic growth of a rifampicin‐marked strain of the seedborne rice pathogen Pseudomonas fuscovaginae. The bacterium was found in most tested seeds indicating that, even without visible discolouration, seed transmission is possible. Crushed discoloured seeds contained more bacterial cells than did non‐crushed discoloured seeds. These bacteria were released during seed soaking, contaminating clean seed and lowering seed germination. Cells of a rifampicin‐resistant strain of P. fuscovaginae, which had been inoculated onto rice seeds, were subsequently recovered from different growth stages and from different rice tissues, thereby indicating endophytic colonization. These results have implications for seedling establishment, as symptomless seeds do not assure disease‐free seeds, and the presence of seedborne bacteria results in poor germination and poor seedling establishment. Elimination of seedborne bacteria by soaking in sodium hypochlorite can increase seed germination. This could be used in developing control strategies, and, if practised regularly, reduce entry of seedborne disease‐causing organisms into crops, resulting in lower disease pressure.     

The ecological life cycle of Thlaspi perfoliatum and a comparison with published studies on Thlaspi arvense

Thlaspi perfoliatum L. is an introduced winter annual that grows in waste places, but it is not a troublesome agricultural weed. Seeds are dormant at maturity in spring, afterripen during summer and germinate in autumn. Non-dormant seeds that fail to germinate in autumn are induced into secondary dormancy by low winter temperatures and cannot germinate the following spring. These seeds afterripen during summer and germinate in autumn if conditions are suitable. Vernalization is not an absolute requirement for flowering. However, it shortens the time to flowering, and vernalized plants produce many more flowers and seeds than non-vernalized plants. Results from studies on seed germination and flowering of T. perfoliatum are compared to published studies on the life cycle of Thlaspi arvense L., an introduced species that is a serious agricultural weed which behaves both as a winter and a summer annual.     

Physiological and Cytological Studies on the Inhibition of Striga Seed Germination by the Plant Growth-promoting Bacterium Azospirillum brasilense

Striga hermonthica (Del.) Benth. is an obligate parasitic weed of tropical cereals whose rhizosphere can also be colonised by bacteria of the genus Azospirillum. A previous study demonstrated that the two organisms (Azospirillum and striga) interacted during cerealroot colonisation. Two strains of A. brasilense isolated from an African sorghum rhizosphere prevented the germination of striga seeds although they were stimulated to germinate by the presence of sorghum roots. Azospirillum cells suspended in a synthetic germination stimulant (GR24) did not inhibit striga seed germination, but did block radicle elongation. Those radicles had an abnormal morphology, and contained no vacuolated cells in the root elongation zone. Lipophilic compounds extracted from the medium of bacteria in the log and stationary growth phases prevented the germination of striga seeds.     

Seed germination, seedling establishment and growth patterns of wrinklegrass (Ischaemum rugosum Salisb.)

Several laboratory and glasshouse experiments were conducted to assess seed germination, seedling establishment and growth patterns of wrinklegrass (Ischaemum rugosum Salisb.) influenced by temperature and light regimes, and chemical media. Wrinklegrass was a positively photoblastic species, and seed germination was temperature‐dependent and light‐mediated. Seeds soaked in distilled water for 24 h, or oven‐dried at the respective temperature regimes of 15, 20, 25, 30, 35, or 40°C prior to treatment in distilled water and incubated in darkness, failed to germinate. Likewise, no germination prevailed when the seeds were exposed to similar temperature regimes and treated with 0.2 m KNO₃, 5% H₂O₂ or 0.01 m HNO₃, and incubated under continuous darkness. Seeds treated with 5% H₂O₂ at 30°C, or oven‐dried and treated with 0.01% M HNO₃ at 35°C registered 10 and 20% germination. Approximately 75 and 90% of the light‐exposed seeds for all treatments germinated in the first three and six days at 25°C. No germination occurred at 15°C in the first three days after treatment. Seeds subjected to 40°C for six days after treatment recorded 36% germination. The optimum temperatures for seed germination were 25–30°C. Seed drying and soaking treatments widened the windows of the optimal temperatures for wrinklegrass germination. The acidic media of KNO₃, H₂O₂ or HNO₃ favored seed germination. Less than 5% of seed germination occurred with burial or water inundation at depths exceeding 2 cm. Seed burial or inundation at ≥2 cm depths inhibited seed germination. Seeds sown onto moist paddy soils registered ca. 50% germination. Free‐floating seeds on the water surface registered ca. 98% germination within the first six days after seeding. The mean number of seedlings that survived was inversely proportional to water depths, with close to 100% mortality at the 14 cm depths of inundation. Both plant height and seedling survival were linearly proportional to the amount of root mass of seedlings which penetrated the soil. The weed was a prolific seed producer (ca. 6000 seeds/genet or 18 000 seeds/genet per year). The vegetative and reproductive efforts of each wrinklegrass plant registered values of 0.68 and 0.32, respectively.     

Seed heteromorphism in carolina dayflower (Commelina caroliniana Walter)

Carolina dayflower (Commelina caroliniana Walter), infesting soybean (Glycine max L.) fields on northern Kyushu Island, Japan, has seed heteromorphism; that is, it produces two types of seeds: pericarp and naked. However, there is no information about their germination behavior. The purpose of this study was to understand the germination characteristics of carolina dayflower seeds and to clarify the difference between the pericarp and naked seeds. On the shape of the seed, the pericarp seeds were significantly longer than the naked ones, with no significant difference in width or thickness. Both the pericarp and the naked seeds could germinate at >20°C, and at 30°C, their cumulative germination rate at 7 days after sowing was the highest, at >90%. Light had no effect on seed germination. The cumulative germination rate after 7 days, when the seeds had been stored dry, wet or under water at a low temperature, was significantly lower than after storage at room temperature, suggesting that a higher temperature and concentration of oxygen during the seed‐storage period affects the germination of carolina dayflower. However, there was no difference in the germination behavior between the pericarp and the naked seeds.     

Influence of Root Exudates on Orobanche Seed Germination*

Broomrape (Orobanche ramosa L.) is a common root parasite of solanaceous, leguminous and other crops grown in the semi-arid regions of the world. The seeds germinate when root exudates from host plants are released in their immediate vicinity (Lindley, 1853; Koch, 1887; Chabrolin, 1934). Brown et al. (1951a) reported that non-host plants, such as flax (Linum usitatissimum L.) may stimulate Orobanche seed germination without being parasitized. The stimulating properties of flax exudate were studied by Brown et al. (1951b). They reported that the stimulant was unstable in alkaline solutions, but moderately stable in weakly acidic media, which may indicate the presence of an acidic (lactone) grouping. Nash & Wilhelm (1960) reported that gibberellic acid in agar media stimulated O. ramosa seed germination. Abu- Shakra, Miah & Saghir (1970) found that pre-treatment of 0. ramosa seeds with 100 ppm of gibberellic acid followed by incubation on a flax-root diffusate agar medium gave a high (81·7%) germination. The purpose of this study was to collect root exudates from three species of plants cultured under three experimental systems, namely (a) germ-free, (b) glasshouse (non-sterile), and (c) growth chamber (hydroponic, initially aseptic), and to evaluate their biological activity as germination stimulants for O. ramosa seeds. The plants used were tomato (Lycopersicon esculentum Mill.), sorghum (Sorghum vulgare Pers.) and flax. Exudate from marigold (Tagetes erecta L.) also was collected from germ-free culture.     

Moisture content and dormancy in Striga asiatica seeds

Freshly harvested Striga asiatica L. seeds will germinate in response to a stimulant only after the passage of time, an after-ripening period, and exposure to moisture at a suitable temperature, a conditioning period. To investigate the role of seed moisture content in the regulation of the after-ripening period, seeds were placed in chambers having specific relative humidity of 6%, 14%, 33%, 75% and 91% for 30, 60, 90 and 150 days. The seeds were then conditioned and germination percentage, response to tetrazolium and seed moisture contents were measured. Seeds at moisture contents less than 10% at the start of conditioning had germination of greater than 93%. Seeds at moisture contents over 10% at the start of conditioning could germinate between 60% and 3%, with germination decreasing as seed moisture content at the start of conditioning increased. The highest moisture content (17%) and lowest germination percentage (3%) occurred in seeds stored at 91% relative humidity for 150 days. There was a linear relationship of a high degree of correlation (0.997) between a positive tetrazolium test and germination capacity. Germination capacity of seeds could be changed from 90% to 3% by prolonged storage in water (dilute benomyl solution), causing `wet dormancy', then returned to 90% germination by returning to dry storage. Seed moisture content at the beginning of conditioning appears to control the responsiveness of the seeds to germination stimulants. The implications of these findings to the control of the parasite are discussed.     

Dormancy and viability of Phalaris minor seed in a rice–wheat cropping system

Seed placement, soil temperature and soil moisture content influenced the process of after-ripening in Phalaris minor seeds. Seeds of P. minor collected from the soil just after wheat harvesting exhibited higher germination than seeds from P. minor threshed directly. There was a pronounced impact of periodic inhabitation of seed into the soil on germination after its dispersal. Germination was strongly inhibited when the seed was kept in soil at more than field capacity (FC) or in water. Maximum germination of seed incubated in soil at FC occurred at 30°C while a temperature of 40°C favoured after-ripening of seed when mixed with dry soil or kept dry without any medium. Release from conditional dormancy was quicker in the seed retrieved from the soil kept at 20°C than at 10°C. Seed release from conditional dormancy and germination increased with a rise in temperature from 30 to 40°C when the seed was retrieved from incubation in soil at FC for 70 days. The seed kept immersed in water was least responsive to a rise in temperature. Seed recovered from dry soil, or kept without any medium, responded quickly at both temperatures. Light enhanced the germination of Phalaris minor seed. The seedbank subjected to rice (Oryza sativa) field management conditions lost vigour in comparison with the seed stored in laboratory. There was significant variability in seed viability when exposed to differential water management conditions in rice.     

应用免疫金探针定位和定量测定稻种中的水稻细菌性条斑病菌

本文首次报道用免疫金染色和免疫金银染色对水稻细菌性条斑病病种的带菌部位及带菌量的研究。结果表明:病原细菌分布于谷壳、籽实皮、胚和胚乳,主要集中在谷壳上。谷壳外表带菌量多于内表面。籽实皮外表面由于受谷壳污染较之内表面的带菌量多。籽实皮内表面、胚和胚乳的带菌量均较少,差异不显著。谷壳、谷壳内表面、籽实皮、胚乳(或胚)的带菌量之比为36:13:9:1。     

应用免疫金探针定位和定量测定稻种中的水稻细菌性条斑病菌

 本文首次报道用免疫金染色和免疫金银染色对水稻细菌性条斑病病种的带菌部位及带菌量的研究。结果表明:病原细菌分布于谷壳、籽实皮、胚和胚乳,主要集中在谷壳上。谷壳外表带菌量多于内表面。籽实皮外表面由于受谷壳污染较之内表面的带菌量多。籽实皮内表面、胚和胚乳的带菌量均较少,差异不显著。谷壳、谷壳内表面、籽实皮、胚乳(或胚)的带菌量之比为36:13:9:1。     

Germination ecology of five arable Ranunculaceae species

Germination and emergence are critical life stages for annual plants and so their full understanding is essential for managing arable plant populations. This study investigated the most important species‐specific environmental cues that regulate seed germination and emergence (temperature and light) of the arable Ranunculaceae species Consolida orientalis, Consolida pubescens, Delphinium gracile, Delphinium halteratum ssp. verdunense and Nigella gallica, to propose management strategies for their preservation in agro‐ecosystems. Growth chamber and outdoor pot experiments were conducted for two consecutive seasons to analyse light (complete darkness or 12 h light) and temperature (5/10, 5/15 and 10/20°C) requirements and emergence patterns. The relative light germination requirement (ΔG_light), which extends from ?100 (complete darkness) to 100 (light), was estimated. Weibull functions were fitted to observed emergence (%) in pots. For all species, germination was higher in complete darkness than with a light regime (?60 < ΔG_light < ?95). This dark requirement indicates better germination for buried seeds. A tillage operation just after seed shed is therefore recommended. Consolida spp. germinate and emerge almost exclusively in autumn–winter, while Delphinium spp. and N. gallica can also germinate in spring. These arable plants would be able to adapt to delayed sowings, an important strategy for avoiding early‐emerging competitive weeds. Facultative winter‐germinating species could face early herbicide treatments if sufficient emergence occurs in winter–spring. These results bring new information to help develop conservation strategies for these species in agro‐ecosystems.     

Dormancy,germination and emergence of Urochloa panicoides regulated by temperature

Urochloa panicoides is an annual weed of summer crops. In Argentina, in subhumid areas with monsoon rainfall, it germinates and establishes in a single flush. To (i) identify the environmental factors that modify its seed dormancy level and germination and (ii) quantify the parameters describing the thermal behaviour of the germination and emergence dynamics of this weed under non‐limiting water conditions, we established a set of germination experiments performed (i) under controlled conditions using seeds after ripened for 3 or 6 months in different thermal and hydric conditions and (ii) under field conditions, where the soil temperature was modified by applying different shading levels. Seed dormancy level remained high with 3 months after ripening in all treatments. After 6 months, seeds stored at 4°C in dry conditions did not germinate at any temperature, while seeds stored at 25°C in dry conditions and in situ germinated c. 20% and 60% respectively. Germination percentage was higher in seeds harvested before their natural dispersal. The base, optimum and maximum temperatures for seed germination were 6, 35 and 45°C respectively. Shading reduced the number of emerged seedlings, possibly by reducing the soil thermal amplitude. The results explained the dormancy‐breaking mechanism of U. panicoides that allows a high germination rate in the field when rainfall occurs.     

Germination ecology of Leptochloa chinensis: a new weed in the Italian rice agro-environment

Leptochloa chinensis is a new weed that has been found with increasing frequency in Italian rice paddies. The germination ecology of L. chinensis seeds was studied in order to investigate the development mechanisms and survival strategy of this weed in rice paddies of northern Italy. Leptochloa chinensis seeds showed no dormancy and exhibited germination even in anoxic conditions. Germination was strongly influenced by temperature (minimum around 15°C; optimal 25–35°C) and light (phytochrome dependent). Temperature fluctuation caused an increase of seed germination in the dark. Seed burial also strongly inhibited germination and emergence of this species. At 5 cm seed burial only 5% of seedlings emerged in flooded conditions, while at the same depth, but with no flooding, no seedling emergence was observed. This phenomenon was not due to oxygen depletion, as germination was not inhibited by complete anoxia, as demonstrated by the fact that some seedlings did emerge in flooding conditions when water was no deeper than 6 cm. Seed burial and concomitant flooding induced an unusual germination: first coleoptile emergence and subsequently emergence of the radicle was observed. The possible exploitation of this knowledge for weed management is discussed.     

Seed germination of the invasive species Piper aduncum as influenced by high temperature and water stress

Piper aduncum is a common woody weed throughout the tropics and a successful invader in many areas. As this species most often dominates roadsides and forest margins, and these habitats frequently experience extreme changes in temperature and water availability, it is important to understand how the seeds adapt to these stresses. This study investigated the effect of high temperatures and water stress, continuous, periodic or transient, on P. aduncum seed germination before or after imbibition. It was found that P. aduncum seeds exhibited only intermediate tolerance and did not germinate at temperatures above 35°C or water potentials below ?0.6 MPa. However, this species is well adapted to local conditions, with fruit ripening in the rainy season, seed desiccation tolerance to eRH 8%, rapid germination under wet conditions, higher temperature tolerance than the maximum ground temperature, germination of most seeds after 60 h continuous heat treatment or daily periodic heat treatments up to 5 h at 40°C and insensitivity to 12 h heat treatment at 40°C during the whole germination process. The intermediate tolerance to high temperature and water stress of P. aduncum, in combination with local environmental conditions in Xishuangbanna, makes P. aduncum an intermediate invader in this area.     

Environmental and physiological factors affecting the fate of seeds of yarrow (Achillea millefolium L.) in arable land in New Zealand

Yarrow (Achillea millefolium L.) has arisen as a problem weed in arable land in New Zealand. A spring-sown yarrow population of 58 plants m^?2 produced approximately 243000 seeds. These were capable of immediate germination in the autumn, but virtually no germination occurred in the winter and up to 80–100% at other times of the year. Seeds are positively photoblastic, but a considerable percentage of seed (>30%) may germinate in the dark after chilling, coat pricking, exposure to high nitrate concentrations or to alternating temperatures. Seed buried at 32 cm in undisturbed soil lost viability slowly over 4 years while seed in the top 8 cm lost viability almost completely within 2 years.     

Seed biology of Mikania micrantha in Viti Levu,Fiji

Mikania micrantha (mile‐a‐minute) reproduces both by seed and vegetatively. A study to determine the possible pollinators, seed production rates, temperature and salinity limitations to germination and its seedbank size and persistence was conducted in Viti Levu, Fiji. Representatives of the Diptera had the greatest percentage of all floral visits (38%), followed by Hymenoptera (34%) and Lepidoptera (27%), while the honeybee was the most recurring visitor (18% of all visits). Flower heads (capitula) within the inflorescence commonly formed four viable seeds, resulting in 60 820 filled seed being produced per m². However, the seedbanks formed were not massive (600 seed m^?2) and they were moderately persistent (T₅₀: 1–3 years). Seed germination from both high and moderate rainfall regions occurred rapidly, under a wide range of temperature regimes, with no primary dormancy being observed. This study indicates that the seed reproductive success of M. micrantha in the two rainfall regions of Fiji is due to a number of factors, including the production of large numbers of flowers, successful pollination by local insects and the subsequent production of a large number of viable seeds. These seeds have high viability, no dormancy and are capable of forming small‐to‐medium seedbanks that are moderately long‐lived. In addition, seeds can germinate under a wide range of temperature and salinity conditions. This knowledge on seed production, biology, longevity and salt tolerance is vital in the development of management plans of M. micrantha in Fiji.     

Epidemiology of dark leaf spot caused by Alternaria brassicicola and A. brassicae in organic seed production of cauliflower

In organic seed production of Brassica vegetables, infections by Alternaria brassicicola and A. brassicae can cause severe losses of yield and seed quality. Four field experiments with or without artificial inoculation with A. brassicicola were conducted in organically managed seed‐production crops of cauliflower cv. Opaal RZ in 2005 and 2006 in the Netherlands. The development of A. brassicicola and A. brassicae on pod tissues and developing seeds was followed and seed quality was assessed. Alternaria brassicicola was externally present on 1·2% of the seeds 14 days after flowering and observed internally within 4 weeks after flowering. In both seasons, seed colonization by the pathogen increased slowly until maturation but sharply increased during maturation. A similar pattern was found for the colonization of pod tissues by A. brassicicola as quantified by TaqMan‐PCR. The incidence of A. brassicicola on mature seeds reached 70–90%. Internal colonization was found for 62–80% of the seeds. External and internal seed colonization by A. brassicae was much lower, with incidences below 3%. The quality of harvested seeds was generally low, with less than 80% of seeds able to germinate. Seed quality was not affected by warm water treatments. It was concluded that A. brassicicola and A. brassicae have the potential to infect pods and seeds soon after flowering. For the production of high quality seeds, producers must prevent such early infections. Therefore, new control measures are needed for use in organic cropping systems.     

Influence of after-ripening environments on the germination characteristics and seed fate of Italian ryegrass (Lolium multiflorum)

Italian ryegrass ( Lolium multiflorum Lam.) is an exotic winter annual weed that recently has seriously infested wheat fields in central Japan. To understand the influence of tillage systems on the germination ecology and seedbank dynamics of naturalized Italian ryegrass, we compared the fate of the seeds that were after-ripened in the soil with that of the seeds on the soil surface, and then examined the germination characteristics of the seeds retrieved from both treatments by germination tests in both a light/dark (12 h photoperiod) condition and in constant darkness. The seedling emergence was greater initially for the seeds after-ripened on the soil surface than for the seeds in the soil. The component of emergent seeds after-ripened on the soil surface increased as time passed during the summer, whereas that of emergent seeds in the soil scarcely increased. This contrasting emergence was, in part, attributable to the ability of the seeds to germinate at higher temperatures in the light/dark condition than in constant darkness. These results suggest that untilled systems, which leave many seeds on the soil surface, might promote the rapid depletion of Italian ryegrass seeds through germination before wheat sowing in early winter. In contrast, seed burial by tillage might inhibit the germination of the seeds and form a large transient seed bank available to germinate during the wheat-growing season.     

Comparison of extraction procedures for detection of Xanthomonas axonopodis pv. phaseoli in common bean seed

Xanthomonas axonopodis pv. phaseoli (Xap) is an important seedborne pathogen of Phaseolus vulgaris. Accurate seed health testing methods are critical to protect seed quality and meet phytosanitary requirements. Currently employed selective media‐based methods include several variations in extraction procedures. In order to optimize pathogen extraction from seeds, the influence of different extraction steps on the sensitivity of Xap detection was assessed. Seeds were inoculated by vacuum infiltration with Xap to achieve inoculum levels from 10¹ to 10⁵ CFU per seed; one contaminated seed was mixed into 1000‐seed subsamples of uncontaminated P. vulgaris seeds. Thirty subsamples of 1000 seeds were tested using each different extraction procedure. These included soaking whole seeds in sterilized saline phosphate buffer, either overnight at 4°C or for 3 h at room temperature, with or without vacuum extraction, and either with or without concentrating the seed extract by centrifuging. Seed extract dilutions were cultured on semiselective agar media MT and XCP1. The percentages of positive subsamples were compared to measure the effects of each extraction step on detection sensitivity. Vacuum extraction and centrifugation of seed extracts increased sensitivity; the highest sensitivity was obtained with the 3 h vacuum extraction followed by centrifugation. These results were confirmed with naturally infested seeds; Xap was detected in 48 of 70 samples using the 3 h vacuum extraction with centrifugation, whereas only 35 of 70 field samples tested positive using overnight soaking, a significant difference. The results suggest that these steps would be valuable modifications to the current method approved by the International Seed Testing Association (ISTA).     

Burial effects on seed germination and seedling establishment of Prosopis juliflora (SW.) DC

Seed germination and seedling establishment are critical stages in plant life history, and both processes may be affected when buried under soil. Our aim was to investigate the effects of soil burial on the germination and seedling establishment of invasive Prosopis juliflora (Fabaceae). We studied the effects of burial, from soil surface to 4?cm deep, under greenhouse conditions and the effects of seed-storing in burial conditions at two depths (5 and 20?cm) and two storing periods (during 6 and 12?months) in field conditions. Soil burial limited the germination and the establishment of P. juliflora, which did not germinate at soil depths deeper than 3?cm. Some seedlings coming from seeds germinated at 2?cm, and every seedling at the soil surface and at 3?cm deep died. In contrast, survivorship of emerged seedlings was close to 90% between 0.5 and 2?cm depth. Seedlings developed the longest shoots at 1?cm deep, indicating that P. juliflora was able to get advantage at a moderate soil depth. Un-embedded seeds survived buried in the soil for months; however, their germination and the emerged seedling survivorship decreased after burial for 12?months.