Lantana camara and the fruit- and seed-damaging fly,Ophiomyia lantanae (Agromyzidae): Seed predator,recruitment promoter or dispersal disrupter?

The seed damaging fly, Ophiomyia lantanae is a widespread agent that infests the fruits and seeds of Lantana camara. Its effectiveness in reducing seed dispersal or seedling recruitment has not been comprehensively tested. We determined the effects of O. lantanae damage on fruit removal rates of L. camara in a field study. We also investigated recruitment responses (seed germination in growth cabinets and seedling emergence in pot trials) of two common L. camara biotypes, pink and pink-edged red (PER) to O. lantanae damage. We found that fewer damaged fruits were removed than undamaged fruits, suggesting that frugivores select against damaged fruit. In all recruitment experiments, the responses to damage were biotype dependent. In germination experiments, these differential responses were primarily due to significantly reduced germination of damaged PER seeds. Seedling emergence from pots of the PER biotype was also lower in damaged seeds (27.5%) than undamaged seeds (56.1%). In contrast, emergence increased with seed damage in the pink biotype, from 28.2 to 41.1%. O. lantanae damage significantly reduced seed mass only in the smaller seeded PER biotype. In the larger seeded pink biotype, it may have promoted seedling emergence by interfering with dormancy mechanisms. We also examined in situ seed banks at L. camara populations prior to peak seed production. Mean viable seed densities ranged from 78.6 to 402.8 seeds m⁻². Of these, non-dormant seeds comprised 24.6 to 98.2 seeds m⁻², suggesting that recruitment is unlikely to be seed limited. We conclude that while O. lantanae influences recruitment and dispersal processes, the magnitude of the responses measured are unlikely to greatly influence plant densities in south-east Queensland infestations.     

Temperature and rate of seed consumption by ground beetles (Carabidae)

Predation is a key source of seed mortality in many weed species and thus is a part of natural control. In the field, the intensity of seed predation by invertebrates varies during the course of a year. One source of this variation is fluctuations in ambient temperature. Here, the effect of temperature on seed consumption is investigated for the first time, using two abundant carabid seed predators, Pseudoophonus rufipes and Harpalus affinis (Coleoptera: Carabidae), and dandelion (Taraxacum officinale) as a model system. Field collected individuals were sexed, kept at one of six constant temperatures between 10 and 28 °C and provided with a surplus of seed. Seed consumption was recorded over a period of 4 days. Averaged over all the temperatures, the smaller H. affinis consumed 12.2 seeds day^?1 and larger P. rufipes 29 seeds day^?1. On average, females consumed more seeds than males. Seed consumption by both species increased with temperature. In H. affinis the increase was linear and different for males and females. In P. rufipes the consumption was similar in both sexes but curvilinear because there was no further increase in consumption above 20 °C. Assuming a linear relationship between temperature and consumption at up to 20 °C we calculated the temperature at which seed consumption ceased (?0.1 to 0.3 °C in H. affinis and 6.3–6.9 °C in P. rufipes) and the increment in seed consumption per 1 °C increase in temperature above this threshold (0.4–1.0 and 1.5–4.2 seeds individual^?1 day^?1, respectively) for the two species. Thus, it is possible to calculate the average daily consumption of each species over a range of temperatures up to 20 °C.     

Density and distribution of seeds in bottom sediments in Zostera marina beds in Ago Bay,central Japan

The density of Zostera marina L. seeds in bottom sediments was examined to study the reproductive patterns of the Z. marina population in Ago Bay, Mie Prefecture, central Japan.Seeds and seed coats were numerous in Tategami, where the annual type of Z. marina grows. In contrast, seeds were scarce in Hamajima, where the perennial type of Z. marina grows. Bottom sediment was sampled with sediment cores at Tategami in November 2004 and March 2005 to examine density and depth distribution of the seeds. Seeds were found as deep as 8 cm, but no deeper. On the other hand, empty seed coats were found as deep as 16 cm in both months. In the upper layers of the sediment to a depth of 8 cm, the average number of seed coats was 7960 ± 2997 m⁻² in November and 16,318 ± 2922 m⁻² in March. Deeper than 8 cm, the number of seed coats gradually decreased owing to decomposition, and none was found below 16 cm. We used the density of reproductive shoots and number of seeds per spadix in Tategami to estimate the fate of seeds and seed coats of the annual type of Z. marina in bottom sediments: out of the 6000 seeds m⁻² produced annually, 72% disappears from the stand and 28% is buried locally. The density and distribution of Z. marina seeds are among the most important factors in maintenance and propagation of the annual population at Tategami.     

Germination responses of Croton macrostachyus (Euphorbiaceae) to various physico-chemical pretreatment conditions

Croton macrostachyus Hochst. ex Del. (Euphorbiaceae) is a multipurpose, deciduous, and medium sized tree of pantropic occurrence. Because the species has numerous useful qualities (e.g., establishment and growth in disturbed sites, drought tolerance, fast growth rate, copious litter/necromass production, suitability for agroforestry, and ability to attract avian frugivores), its speedy restoration has become increasingly critical. Germination studies were therefore conducted on seeds pooled from five widely located provenances with a view to supporting efforts geared toward the speedy propagation and restoration of this valuable tree species. Seed pretreatments were achieved using various dilution levels of plant-derived smoke–water (1:1, 1:10, 1:100 and 1:1000), as well as gibberellic acid (GA₃) or potassium nitrate (KNO₃) ranging in concentration from 0.1 to 100 μmol. The control was to use distilled water for seed pretreatment. Seeds were germinated under either illuminated (ca 60 μmol m^− 2 s^− 1; cool-white fluorescent lamp) or non-illuminated conditions. Experiments on the impact of seed storage durations, as well as storage temperatures were also conducted. The study found that germination percentage (GP: ca 90%), and mean germination time (MGT: 14 days) were significantly (P < 0.001) better when seeds were pretreated with smoke–water and germinated under non-illuminated conditions, than when these were pretreated with various concentrations of GA₃ or KNO₃ (GP and MGT of ca 65% and 20 days, respectively). Germination percentage (GP) and germination vigor (GV) declined with increasing storage-time for all storage temperatures, but GV's decline was faster for seeds stored at 22 °C than for those stored at 5 and 15 °C. On the other hand, mean germination time (MGT) increased significantly (P < 0.01) with seed storage-time of up to 8 months at 5, 15, and 22 °C, but the increase was more marked for seeds stored at 22 °C than for those stored at 5 and 15 °C. From these investigations, it is concluded that germination of C. macrostachyus seeds through use of smoke–water is faster, cheaper, and technically less demanding, compared to that of either GA₃ or KNO₃. The study also concludes that C. macrostachyus is intermediate between orthodox and recalcitrant seeds, and that it is non-photoblastic.     

The seed ecology of an ornamental wattle in South Africa — Why has Acacia elata not invaded a greater area?

Australian Acacia species introduced to South Africa as ornamentals have notably smaller invasive ranges than those introduced for forestry or dune stabilization. We asked whether the relatively small invasive extent of Acacia elata, a species used widely for ornamental purposes, is due to low rates of reproduction. Age at reproductive maturity, seed dispersal, annual seed production, seed bank dynamics and seed germination and viability were assessed at five sites in the Western Cape. Results indicate that A. elata has similar traits to other invasive Australia Acacia species: annual seed input into the leaf litter was high (up to 5000 seeds m^− 2); large seed banks develop (> 20,000 seeds m^− 2) in established stands; seed germinability is high (> 90%); seeds accumulate mostly in the top soil layers but can infiltrate to depths of 40 cm; and seed germination appears to be stimulated by fire. However the age at the onset of reproduction (~ 4 years) is longer than most widespread invaders (~ 3 years) and dispersal is fairly limited (seeds fell up to distances of 6 m from the parent canopy; the highest density of seed rain was found directly under the canopy with over 20% of seeds falling directly under the terminal branches). We suggest that the current limited distribution of invasive A. elata populations is the result of the relatively small size of initial populations (cf. large plantations and widespread plantings for forestry and dune stabilization species), the species' apparent lack of secondary dispersal vectors, and the planting of trees in gardens and urban settings offer limited opportunities for recruitment, proliferation and spread. The species is, however, increasing in abundance and range. We propose methods to improve management of invasions of the species. Management to reduce seed production of this species through classical biological control, as has been achieved for other Australian Acacia species in South Africa, should be prioritised.     

Effect of yak rumen content treatments on seed germination of 11 alpine meadow species on the Qinghai-Tibetan Plateau

The Qinghai-Tibetan Plateau is located in the ‘Third Pole’ of the world, characterized by a harsh environment. Despite this, the alpine meadow ecosystem have developed over a wide area but serious grassland degradation is threatening the ecological environment on the Plateau. Recruitment of new plants to the population, via germination and establishment, is vital to plant community survival. Previous work on the seeds in this area has mainly focused on community-wide germination strategies, seed germination characteristics and their correlations with seed size and seed mass. However, there have been no studies on the effects of soaking in rumen contents on the plant seed germination characteristics of alpine meadow species. The present study had two main objectives: (i) to determine the effect of fresh rumen content from yaks on seed germination characteristics and seedling growth of species common to the eastern Tibetan Plateau alpine meadow, and (ii) to develop an effective method to enhance seed germination. Seeds of 11 common species were collected together with fresh rumen content from three yaks that grazed there. Seed germination tests were conducted after they had been soaked in rumen content for one of six soaking periods (12, 24, 36, 48, 60 or 72 h). The seeds were incubated under natural light conditions of 8 h light at 25 °C and 16 h darkness at 15 °C, for the germination period of 34 days. The results showed that seed germination and seedling growth were affected by soaking time, seed coat completeness and seed type. After soaking in rumen content, the germination percentages of scarified (peeled or with the seed coat cut through) seeds of some species (true seeds Oxytropis ochrocephala and Medicago ruthenia var inschanica, nutlet Carex enervis, achenes Anemone rivularis and Polygonum sibiricum) and complete seeds of C. enervis, and A. rivularis were improved but the duration of soaking was also important. Seed germination of caryopsis Achnatherum inebrians (a toxic grass) was significantly inhibited by any exposure to rumen fluids. Scarified seeds generally had higher germination percentages than complete ones after treatment, but with the increase in soaking time, germination percentages declined and scarified seeds were more sensitive to the treatment than the complete seeds. After soaking in yak rumen content, the germination indices of scarified M. ruthenia at 12 h treatment, O. ochrocephala and achene Rumex acetosa at 12–24 h treatment, nutlet Kobresia humilis at 24 h treatment, P. sibiricum at 24–48 h treatment, C. enervis at 12–48 h treatment and A. rivularis at 12–60 h treatment were significantly higher than the control (P < 0.05), while the germination indices of complete C. enervis seeds at 12 h and 36 h treatment, and A. rivularis at 12–60 h treatment were significantly higher compared with the control. The germination indices of other species gradually decreased with the increase in soaking time. We concluded that yak rumen digestion could enhance, inhibit or not affect seed germination and seedling growth of the alpine meadow species, which might influence seedling recruitment, interspecific competition, and the plant community structure of the eastern Tibetan Plateau alpine meadow. Overall, yak digestion has a positive effect on alpine meadow seed germination and seed dispersal.     

Multiple regeneration pathways in Spathoglottis plicata Blume — A study in vitro

Asymbiotic germination of immature seeds (embryos), and mature seeds and micropropagation of Spathoglottis plicata were described. Effects of three nutrition media namely, Murashige & Skoog (MS); Phytamax (PM); and Phyto-Technology orchid seed sowing medium (P₇₂₃), two carbon sources such as glucose and sucrose at 2–3% (w/v), two plant growth regulators such as 6-benzylaminopurine (BAP; 0.5–3.0 mg l^− 1) and α-naphthalene acetic acid (NAA; 0.5–2.0 mg l^− 1) and peptone (2.0 g l^− 1) were examined on seed germination, early protocorm development and micropropagation. The maximum germination of mature seeds (95%) was recorded in PM medium supplemented with 2% (w/v) sucrose + 2.0 g l^− 1 peptone. For germination of embryos P₇₂₃ medium supplemented with 1.0 mg l^− 1 BAP proved best. Multiple shoot buds or protocorm-like bodies (PLBs) were produced from stem segments of in vitro raised seedlings. Both direct organogenesis and embryogenesis were observed and the morphogenetic response was initiated by different concentrations and combinations of PGRs. The optimum PGR combination for maximal PLB regeneration was 1.0 mg l^− 1 NAA + 2.5 mg l^− 1 BAP, while 1.0 mg l^− 1 NAA + 1.0 mg l^− 1 BAP for shoot bud development. Strong and stout root system was induced in half strength PM medium supplemented with 0.5 mg l^− 1 IAA. The well-rooted plantlets were transferred to pots containing a potting mixture composed of saw dust, coconut coir, humus, and coal pieces at 1:1:1:2 (w/w) with 80% survival in outside environment and flowered after two years of transfer.     

Seed storage and germination in Kumara plicatilis,a tree aloe endemic to mountain fynbos in the Boland,south-western Cape,South Africa

Seed storage under appropriate conditions is a relatively inexpensive means of safeguarding plant genetic material for ex situ conservation. Post-storage germination trials are used to determine the viability of stored seeds, and hence the efficacy of the particular storage treatment. Kumara plicatilis (= Aloe plicatilis) is a tree aloe endemic to mountain fynbos in the Boland, south-western Cape. The viability and germination behaviour of K. plicatilis seeds were assessed for seeds stored for four and nine months at − 80 °C, 4 °C, 25 °C and under ambient conditions in a laboratory. Seeds were germinated under controlled conditions and germination rates and percentages determined. Ungerminated seeds were tested for viability using tetrazolium salt. Seed viability was not significantly reduced during storage. Seeds stored at − 80 °C for four and nine months exhibited the fastest germination rate overall (both 5.9 ± 0.3 weeks, mean ± S.E.), and slowest was for seeds stored under ambient conditions for four and nine months (both 7.8 ± 0.4 weeks). All seed lots showed similar percentage germination after four months of storage (78.0–90.4%). The highest percentage germination overall was for seeds stored at − 80 °C for four months (90.4%) and the lowest was for seeds kept at 4 °C and − 80 °C for nine months (39.2 and 39.6%, respectively). Respective percentage viability for ungerminated seeds in these two treatments was 82% and 87%, respectively, indicating the induction of secondary dormancy. Induced dormancy triggered by protracted cold temperatures may be an adaptation that enables seeds to survive prolonged extreme conditions that are unfavourable for germination. Further research on the long-term storage of aloe seeds would be beneficial for developing long-term seed storage and germination testing protocols for ex situ conservation.     

Exogenous nitric oxide improves seed germination in wheat against mitochondrial oxidative damage induced by high salinity

Effects of exogenous nitric oxide (NO) on starch degradation, oxidation in mitochondria and K⁺/Na⁺ accumulation during seed germination of wheat were investigated under a high salinity level. Seeds of winter wheat (Triticum aestivum L., cv. Huaimai 17) were pre-soaked with 0 mM or 0.1 mM of sodium nitroprusside (SNP, as nitric oxide donor) for 20 h just before germination under 300 mM NaCl. At 300 mM NaCl, exogenous NO increased germination rate and weights of coleoptile and radicle, but decreased seed weight. Exogenous NO also enhanced seed respiration rate and ATP synthesis. In addition, seed starch content decreased while soluble sugar content increased by exogenous NO pre-treatment, which was in accordance with the improved amylase activities in the germinating seeds. Exogenous NO increased the activities of superoxide dismutase (SOD, EC 1.15.1.1) and catalase (CAT, EC 1.11.1.6); whereas decreased the contents of malondialdehyde (MDA) and hydrogen peroxide (H₂O₂), and superoxide anions (O₂^??) release rate in the mitochondria. Exogenous NO also decreased Na⁺ concentration while increased K⁺ concentration in the seeds thereby maintained a balance between K⁺ and Na⁺ during germination under salt stress. It is concluded that exogenous NO treatment on wheat seeds may be a good option to improve seed germination and crop establishment under saline conditions.     

In vitro seed maturation in Brassica rapa L.: Relationship of silique atmosphere to storage reserve deposition

The Brassica rapa L. silique is a self-contained environment that maintains hypoxia around the developing seeds, and in which carbon dioxide accumulates to very high concentrations (>30,000 ppm). How the silique microenvironment modulates the composition and amount of storage reserves in the seeds is of interest because of the important agricultural role played by canola (B. rapa and Brassica napus) as an oilseed. Because of the small volume and dynamic nature of this microenvironment in Brassica, a standardized system was needed to study the environmental role played in storage reserve deposition. For this purpose we have developed a silique culture system that permits maturation of seed in vitro. Siliques excised from plants just 11 days after pollination complete the ripening of their seeds after 20 days of culture in light (200 μmol/m²/s) on MS medium containing 30 g/l sucrose, 0.25 mg/l BAP, and 0.025 mg/l NAA. Cytochemical localization and biochemical analyses revealed that storage reserves were affected by the in vitro maturation system. Although following a comparable ripening timeline to that occurring on the plant, and producing fully germinable seeds, in vitro maturation resulted in a 40% reduction in seed weight and the mature seeds contained decreased lipid, but increased protein, starch and soluble carbohydrates. To study the internal atmosphere surrounding the seeds, we developed a method to capture silique gases in helium with subsequent quantification of O₂ and CO₂ in the sample by gas chromatography. Analysis of the internal silique atmosphere showed that in vitro siliques provided seeds with a less oxygenated environment than they experience attached to the plant. Carbon dioxide concentrations remained high later into the maturation sequence in vitro than on the plant. When sampling gases from siliques attached to plants, we found multiple samples from the same plant resulted in higher variance than when only a single silique was sampled, suggesting that connection to the plant directly influences internal silique gases. Lower O₂ in the in vitro siliques was correlated with depressed lipid content in their mature seeds, supporting the conclusion that oxygen availability limits lipid accumulation. Previous studies showed how environmental factors influence Brassica embryos grown in tissue culture. These systems fail to preserve the component of metabolic regulation that is enforced by the silique wall tissues. Our in vitro maturation system provides a useful tool for specialized investigations since both the gaseous and hormonal environments can be readily manipulated.     

Montpellier broom (Genista monspessulana) and Spanish broom (Spartium junceum) in South Africa: An assessment of invasiveness and options for management

The legumes (Fabaceae) Genista monspessulana and Spartium junceum are major invaders in several other parts of the world, but not yet so in South Africa. We determine their current distributions in South Africa at different spatial scales, assess population structure (soil seed banks and size at reproduction) evaluate current management activities, and provide recommendations for control (including assessing the feasibility of nation-wide eradication). G. monspessulana occurs at nine localities in three quarter-degree cells, covering a total of 22.7 ha. S. junceum is much more widespread, occurring in 33 quarter-degree cells and is frequently cultivated in private gardens. All naturalised or invasive populations are in disturbed areas, mostly along roadsides. Once established, G. monspessulana and S. junceum accumulate large, persistent soil-stored seed banks, ranging in size between 909 and 22,727 (median 1970) seeds/m² and 0 and 21,364 (median 455) seeds/m² for the two species respectively. Both species resprout vigorously after cutting and stump herbicide application (60% of G. monspessulana and 43% of S. junceum plants resprouted) which necessitates regular follow-ups. We estimate that over 10 years, at a cost of about ZAR 81,000 (1 ZAR = 0.114 US$ as on 6 October 2012), G. monspessulana could be extirpated from South Africa. S. junceum is far more widespread and coupled with low effectiveness of control, abundance of seeds and seed longevity, eradication is unfeasible. We recommend that control methods used for S. junceum be improved to prevent resprouting, and that areas are managed to limit the movement of seeds and avoid further spread and establishment. Further studies are required to understand why these two species have failed to replicate the invasiveness shown in other parts of the world.     

Seed size correlates seedling emergence in Terminalia bellerica

Terminalia bellerica Roxb. (Belleric Myrobalan, Vern. — Baheda, Sanskrit-Vibhitaki, Family: Combretaceae) is among multipurpose tree species in India. The dried pulp of the seeds being used for the preparation of an ancient herbal formulation called Triphala (in Hindi). Seed size is considered a useful attribute for the propagation of valuable trees. The effect of seed size on seedling emergence in T. bellerica was studied under nursery conditions. Emergence of seedlings from large (mean dry weight1.18 ± 0.02 g), medium (0.95 ± 0.03 g) and small seeds (0.76 ± 0.03 g) varied significantly (LSD^Sin p < 0.05 = 4.12, Sin = 0.52). Higher numbers of seedlings emerged from the large seeds compared with medium and small seeds. Seed weight also correlated positively with seedling emergence in T. bellerica (r = 0.967, significant α = 0.01, df = 7). Findings of this study will be useful for mass propagation of T. bellerica and reintroduction of elites in different habitats.     

Unravelling the process of weed seed predation: Developing options for better weed control

Seed predation can cause significant losses of weed seeds in agricultural systems and can, thus, contribute to weed control. The removal of Lolium multiflorum and Vicia villosa seeds by harvester ants, Messor barbarus, and granivorous rodents, Mus spretus, in six cereal fields in NE Spain was separated into three sequential processes, namely (1) the probability of finding a seed cache (cache encounter rate), (2) the percentage of seeds utilized once a seed cache has been found (seed exploitation rate) and (3) seed selection if multiple species are present (preference). Identifying the most important behavioural component and factors that drive it may help to better understand and manage seed predation.Seed cache encounter rate correlated well with overall seed removal rate caused by harvester ants (r² = 0.91), or rodents (r² = 0.82). Once found, seed exploitation rates were high and fairly constant from spring to autumn for harvester ants, and low throughout the season for rodents. Harvester ants removed almost all L. multiflorum seeds from caches found, while the exploitation of V. villosa seeds varied across the season. In the case of rodents, cache encounter rate, but not exploitation rate, could be explained by canopy cover provided by the crop. L. multiflorum seemed to be preferred in early 2007, whereas V. villosa was in 2008.The adoption of no-till or minimum tillage systems together with the establishment of field edge vegetation are likely to encourage seed cache encounter and exploitation rates by both harvester ants and rodents, thus leading to increased weed control in semi-arid cereals.     

Formulations of Bacillus subtilis BY-2 suppress Sclerotinia sclerotiorum on oilseed rape in the field

We are developing a collection of Bacillus strains, isolated from different environments, for use in controlling Sclerotinia sclerotiorum on oilseed rape in China and elsewhere. Strain BY-2, isolated from internal tissues of an oilseed rape root, was demonstrated to be Bacillus subtilis based on biochemical and morphological characteristics and on 16S RNA gene sequence. Photographic evidence from gnotobiotic studies using the lacZ-tagged strain BY-2(pUC18) confirmed that this strain was capable of colonizing internal root tissues. Strain BY-2 did not effectively colonize the ectorhizosphere or the surface of the stems or leaves of oilseed rape when applied in pellet or wrap seed treatment formulations. Populations of BY-2 dropped from 10⁸ CFU seed⁻¹ to 10⁴ CFU g root⁻¹ and ⩽10² CFU g stem⁻¹ or leaf⁻¹ after 60 days. Strain BY-2 was applied as a pellet seed treatment formulation alone, as a spray at flowering alone, and as the pellet seed treatment formulation combined with the spray application in two field trials at the Wuxue location conducted in two consecutive years. These three treatments containing BY-2 provided disease control (disease incidence) and mean seed yield that was similar to the chemical control treatment and significantly greater than the pellet without bacteria and non-treated control treatments. All three of these BY-2 treatments were similar to each other with regard to these two metrics and to treatments containing B. subtilis Tu-100, a genetically distinct strain previously shown to be effective against this disease. In two additional field trials, conducted in consecutive years at the Wuhan location, strain BY-2 applied as a wrap seed treatment formulation alone, as a spray at flowering alone, and as the wrap seed treatment combined with the spray application provided disease control (disease incidence) and mean seed yield that was similar to the chemical control treatment. These three BY-2 treatments also were significantly greater than the non-treated control treatments and compared favorably to treatments containing Tu-100. There was no evidence of BY-2 promoting growth of oilseed rape when applied in the pellet or wrap seed treatment formulations in field trials conducted at the Wuxue or at the Wuhan locations. We now have three Bacillus strains (B. subtilis strains BY-2 and Tu-100, B. megaterium A6) that control S. sclerotiorum on oilseed rape in the field that can be tested in strain combinations for enhanced disease control. We also have multiple methods for application of Bacillus strains as both seed treatment and foliar applications were effective.     

Germination response and viability of an endangered tropical conifer Widdringtonia whytei seeds to temperature and light

The tropical conifer Widdringtonia whytei Rendle is an endangered species endemic to Mulanje Mountain in Malawi. A study was conducted for the first time under controlled conditions in order to assess the effects of temperature and light on germination and viability of W. whytei seeds. Seeds incubated at a constant temperature of 20 °C attained the highest cumulative germination percentage (100%) followed by 87% germination under fluctuating temperatures of 15 °C night/25 °C day. No seed germination occurred at temperatures below 15 °C. Seeds that failed to germinate at temperatures below 15 °C showed the highest (> 90%) viability compared to the seeds incubated at 25 °C (60%). Across temperature regimes, germination was significantly higher under light (44.7%) than dark (35.6%) conditions. It is concluded that temperature is one of the critical factors for germination of W. whytei seed. The ability of W. whytei seeds to germinate both in light and darkness implies that the species would unlikely form a persistent soil seed bank, an attribute which is common in species that survive in habitats frequently disturbed by fires.     

Synergistic effects of essential oil-based cream formulations against Culex quinquefasciatus Say and Aedes aegypti L. (Diptera: Culicidae)

Mosquitoes are major arthropod vectors responsible for several pathogenic diseases. In recent years, repellents of botanical origin, particularly essential oils, have been used against mosquitoes and have been found effective and safe. In this study, five different repellent cream formulations (CF1–5) were prepared using combinations of essential oils, including camphor, cinnamon, citronella, lemongrass, lime, orange, neem, basil, Vitex, Lantana, eucalyptus, and clove, and their repellency was tested using Culex quinquefasciatus Say and Aedes aegypti L. under laboratory conditions and compared to the standard synthetic repellent N,N-diethyl-meta-toluamide (DEET-12%, w/w). Among the five cream formulations, CF2 at a dose of 5 mg/cm² showed the longest protection time of 4.18 h and 3.31 h against C. quinquefasciatus and A. aegypti, respectively, under laboratory conditions. CF3 at a dose of 5 mg/cm² was moderately effective, with protection times of 3.42 h and 2.58 h against C. quinquefasciatus and A. aegypti, respectively, under laboratory conditions. CF2 at a dose of 5 mg/cm² was also tested in the field against wild mosquitoes for 3 h, and 100% protection was observed for the entire study period. Thus, CF2 could be used in developing an effective natural repellent as an alternative to the existing synthetic repellents to C. quinquefasciatus and A. aegypti.     

Bacterial decolorization and degradation of the reactive dye Reactive Red 180 by Citrobacter sp. CK3

A bacterial strain, CK3, with remarkable ability to decolorize the reactive textile dye Reactive Red 180, was isolated from the activated sludge collected from a textile mill. Phenotypic characterization and phylogenetic analysis of the 16S rDNA sequence indicated that the bacterial strain belonged to the genus Citrobacter. Bacterial isolate CK3 showed a strong ability to decolorize various reactive textile dyes, including both azo and anthraquinone dyes. Anaerobic conditions with 4 g l^?1 glucose, pH = 7.0 and 32 °C were considered to be the optimum decolorizing conditions. Citrobacter sp. CK3 grew well in a high concentration of dye (200 mg l^?1), resulting in approximately 95% decolorization extent in 36 h, and could tolerate up to 1000 mg l^?1 of dye. UV–vis analyses and colorless bacterial cells suggested that Citrobacter sp. CK3 exhibited decolorizing activity through biodegradation, rather than inactive surface adsorption. It is the first time that a bacterial strain of Citrobacter sp. has been reported with decolorizing ability against both azo and anthraquinone dyes. High decolorization extent and facile conditions show the potential for this bacterial strain to be used in the biological treatment of dyeing mill effluents.     

d-Xylose absorption test: A tool for the assessment of the effect of anticoccidials on the intestinal absorptive capacity of broilers during experimental coccidiosis

Coccidiosis in chickens causes intestinal mucosal lesions and disrupts its integrity leading to a disturbance in absorption of dietary components. The d-xylose absorption test is a sensitive tool of measuring the absorption capacity of the intestine in diseased chickens. In an experiment on broilers, the influence of different anticoccidials on the intestinal absorption capacity of the birds challenged with experimental coccidiosis was evaluated, using the d-xylose absorption test. The experiment had 5 groups of 10 Ross male broiler chickens (24-days-old) as follows: Group 1— negative control received no Eimeria oocystes, Group 2—positive control challenged with mixed Eimeria oocystes, Group 3—positive control dosed with an attenuated oral coccidiosis vaccine, Group 4—positive control dosed with 25 ppm toltrazuril in drinking water and Group 5—positive control received 66 ppm salinomycin sodium, in the diet. The d-xylose absorption test was carried out 5 days after the coccidial infection. Results showed that coccidiosis highly reduced the plasma d-xylose peak level of Group 2 when compared with Group 1 (31 mg/dl at 90 min versus 50 mg/dl at 30–60 min after the d-xylose administration, respectively). The concentration of d-xylose followed cubic (P<0.001, r² = 0.886) and quadratic (P<0.001, r² = 0.686) correlations with time in Group 1 and 2, respectively. Anticoccidials enhanced the uptake of d-xylose in the infected birds. The plasma d-xylose reached to its peak in Group 3, 4, and 5 (38.9, 50 and 47.0 mg/dl, respectively) at 60–90 min after the d-xylose administration and had quadratic functions with time (r² = 0.802, 0.883 and 0.860, respectively, P<0.001). The d-xylose absorption test was a sensitive test for evaluating the influence of anticoccidials on the absorption capacity of intestinal mucosae during coccidiosis in broiler chickens.     

High seed Mn content does not affect germination of in vitro produced Centaurium pulchellum seeds

The effect of high Mn²⁺ content on Centaurium pulchellum seed germination has been investigated. Seeds containing extremely high Mn²⁺ content were produced by culturing single-node flowering explants for 2 months in the MS-media, supplemented with Mn in concentrations ranging from 1 to 10,000 μM. Although the seeds displayed the capacity to accumulate high amount of Mn, their germination was undisturbed. EPR spectroscopy was used to measure the ratio of free (aqueous) Mn to bound Mn and it was found that over 97% of total Mn was in the bound form. With elevating the external Mn supply, seed Mn concentration also increased, but the proportion of free Mn²⁺ fraction decreased from 3% in the control (1 μM Mn) to 0.35% and 0.15% in high Mn supply (1000 μM and 10,000 μM, respectively). These results suggest that an elevation of internal Mn concentration in seeds is associated with increased Mn binding pools, hence Mn remains bound during germination. Consequently, the action of potentially harmful Mn²⁺ ions, which may generate ROS and affect seed viability, is alleviated.