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.     

Mating system,outcrossing distance effects and pollen availability in the wind-pollinated treeline species Polylepis australis BITT. (Rosaceae)

Fragmentation may negatively affect plant fitness through pollen limitation and increased levels of inbreeding. Effects of fragmentation may vary with regard to life form and breeding system, and few studies exist for wind-pollinated trees. We examined the effects of hand-selfing, varying outcrossing distances and pollen addition on seed mass and germination rate of Polylepis australis BITT. (Rosaceae), a wind-pollinated treeline species endemic to Argentina. We also investigated pollen germination on the stigma and pollen tube growth to determine compatibility resulting from selfing and outcrossing. Selfing reduced seed germination rates with significant differences between open pollination and outcrosses at 30 km. In addition, we found a tendency for pollen germination and pollen tube growth to decrease following selfing. Between-fragment crosses resulted in a trend of higher reproductive output than within-fragment crosses, whereas values were similar between open pollination and between-fragment crosses. Pollen addition did not increase reproductive success neither in small nor in larger fragments. Our results suggest that highly isolated P. australis forests have a potential for inbreeding depression through selfing and within-fragment crosses. However, the results also indicate that pollen flow between P. australis forest fragments is still effective at the current fragmentation level, counteracting negative effects on seed quality resulting from reproductive isolation.     

Optimisation of pollen viability tests for Acacia podalyriifolia and two ploidys of Acacia mearnsii

Acacia mearnsii (black wattle) is a commercially important forestry species in South Africa, grown for its timber as well as its bark. It is, however, also considered to be an alien invader of indigenous vegetation and for this reason the production of a sterile variety would be highly desirable for commercial forestry in South Africa. Previous research on crosses between diploid and tetraploid parent plants to produce triploid progeny has resulted in poor seed set. One possible barrier preventing seed set could be the viability of the pollen used in the cross pollination operations. Thus a study was conducted to test the pollen viability. In vitro agar media germination tests (ACIAR and Brewbaker and Kwack media) were optimised on Acacia podalyriifolia pollen and then used together with vital stain tests (Sigma® DAB peroxidase and p-phenylendiamine) to test pollen germination and viability of A. mearnsii pollen. These were then compared to in vivo pollen germination on the stigma, and were conducted on both diploid and tetraploid pollen mixes. Results showed that the vital stain tests gave significantly (p < 0.05) higher pollen viability than the agar germination tests and were more in agreement with the results from the pollen germination rate on the stigma. For both the diploid and tetraploid pollen mixes tested, there were no significant differences (p > 0.05) between the two agar media germination tests and between the two vital stain tests.     

The seed bank in a subtropical freshwater marsh: implications for wetland restoration

Seed bank samples were collected from Huli Marsh, a subtropical shallow water mountainous marsh in Hunan Province, South China. Core samples were divided into upper and lower layers (each 5 cm in depth) and allowed to germinate in three water levels (0, 5 and 10 cm) over a 4-month period. A total of 51 species germinated and the mean density was 9211 ± 7188 seedlings m⁻². In the top 5 cm 41 species and 5747 ± 5111 seedlings m⁻² germinated, whereas 40 species and 3464 ± 3363 seedlings m⁻² did so from 5–10 cm. Germinated seedling density was significantly higher in the upper layer, largely due to differences in eight species. With increasing experimental water depth, less seedlings germinated: respectively, 9788 ± 7157 m⁻², 2050 ± 2412 m⁻² and 1978 ± 2616 m⁻², of 44, 21 and 19 species, submerged under 0, 5 or 10 cm. Seven species could emerge only in 0 water level. Vallisneria natans occurred only in 5 cm water, whereas Ottelia alismoides occurred in 10 cm water. In the vegetation survey of the marsh, 25 species were recorded, which was less than half of the species recorded in the seed bank. The top 10 dominants in the standing vegetation, accounting for 89% of vegetation abundance, represented only 10% in the seed bank. Twenty germinated species that also occurred in the standing vegetation accounted for 56% of the total seed bank. Our observed number of species germinating from a Chinese wetland seed bank is within the range observed elsewhere in the northern hemisphere (15–113 species).     

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.     

Impact of temperature on olive (Olea europaea L.) pollen performance in relation to relative humidity and genotype

Olive varieties ‘Koroneiki’, ‘Kalamata’, ‘Mastoidis’ and ‘Amigdalolia’ were employed in two experiments for 3 years to assess the effect of temperature on olive pollen germination and tube growth in relation to relative humidity and genotype. Pollen samples were subjected to pre-incubation at 10, 20, 30 or 40 °C in combination with decreased air relative humidity – 80, 40, 30 or 20%, respectively – for 24 h to simulate temperature stress that is observed during pollen dispersal; and subsequently in vitro cultured. In the second experiment, pollen was exposed at 15, 20, 25 and 30 °C for 24 h in vitro to evaluate pollen response in conditions of water and nutrients availability and to determine the optimum pollen germination and tube growth temperatures for each cultivar. The highest pre-incubation temperature treatment (40 °C) prevented pollen germination in ‘Koroneiki’ and ‘Mastoidis’, with the less affected varieties (‘Amigdalolia’ and ‘Kalamata’) having average germination percentages of only 7.6 and 2%, respectively. Pre-incubation at 30 °C had a negative impact on pollen germination in ‘Koroneiki’ (?65%), ‘Kalamata’ (?20%) and ‘Amigdalolia’ (?72%) compared to the control (20 °C). Pollen pre-incubation at 40 °C decreased significantly the pollen tube length in ‘Kalamata’ (?50%) and ‘Amigdalolia’ (?52%). In the second experiment, in vitro pollen germination increased after incubation at 25 °C for ‘Koroneiki’ (+6%), ‘Mastoidis’ (+52%), ‘Kalamata’ (+10%) and ‘Amigdalolia’ (+10%) compared to the control (20 °C). At 30 °C germination percentages for ‘Mastoidis’, ‘Kalamata’ and ‘Amigdalolia’ were 8, 6 and 14% higher, respectively, compared to the control (20 °C). Pollen tube length also increased with incubation temperature for all of the studied cultivars. Based on the cumulative stress response index (CSRI) that was calculated for high temperature stress the varieties were classified: ‘Mastoidis’ and ‘Kalamata’ as tolerant and ‘Koroneiki’ and ‘Amigdalolia’ as intermediate at 30 °C while all studied cultivars were sensitive at 40 °C. The observed strong genotype-differentiated response in high and low temperature stress could be exploited by plant breeders towards producing new tolerant olive varieties.     

Differentiation between Neotropical rainforest,dry forest,and savannah ecosystems by their modern pollen spectra and implications for the fossil pollen record

Accurate differentiation between tropical forest and savannah ecosystems in the fossil pollen record is hampered by the combination of: i) poor taxonomic resolution in pollen identification, and ii) the high species diversity of many lowland tropical families, i.e. with many different growth forms living in numerous environmental settings. These barriers to interpreting the fossil record hinder our understanding of the past distributions of different Neotropical ecosystems and consequently cloud our knowledge of past climatic, biodiversity and carbon storage patterns. Modern pollen studies facilitate an improved understanding of how ecosystems are represented by the pollen their plants produce and therefore aid interpretation of fossil pollen records. To understand how to differentiate ecosystems palynologically, it is essential that a consistent sampling method is used across ecosystems. However, to date, modern pollen studies from tropical South America have employed a variety of methodologies (e.g. pollen traps, moss polsters, soil samples). In this paper, we present the first modern pollen study from the Neotropics to examine the modern pollen rain from moist evergreen tropical forest (METF), semi-deciduous dry tropical forest (SDTF) and wooded savannah (cerradão) using a consistent sampling methodology (pollen traps). Pollen rain was sampled annually in September for the years 1999–2001 from within permanent vegetation study plots in, or near, the Noel Kempff Mercado National Park (NKMNP), Bolivia. Comparison of the modern pollen rain within these plots with detailed floristic inventories allowed estimates of the relative pollen productivity and dispersal for individual taxa to be made (% pollen/% vegetation or ‘p/v’). The applicability of these data to interpreting fossil records from lake sediments was then explored by comparison with pollen assemblages obtained from five lake surface samples.Pollen productivity is demonstrated to vary inter-annually and conforms to a consistent hierarchy for any given year: METF > SDTF > cerradão. This suggests an association between pollen productivity and basic structural characteristics of the ecosystem, i.e. closed canopy vs. open canopy vs. savannah. Comparison of modern pollen and vegetation revealed that some important floristic elements were completely absent from the pollen: Qualea and Erisma (METF), Bauhinia, Simira and Guazuma (SDTF), and Pouteria and Caryocar (cerradão). Anadenanthera was found to be abundant in both the pollen and flora of SDTF (p/v = 3.6), while Poaceae was relatively poorly represented in cerradão (0.2). Moraceae, Cecropia and Schefflera were found to be over-represented palynologically in all ecosystems. Overall, the data demonstrated that no one taxon could be used as a definitive indicator of any of the ecosystems. Instead, associations of taxa were found to be important: METF = Moraceae (> 40%), Cecropia, Hyeronima, Celtis; SDTF = Anadenanthera, Apuleia, Ferdinandusa and non-arboreal Asteraceae, Bromeliaceae, Piper and fern spores; cerradão = Poaceae, Myrtaceae, Borreria, Solanum plus Asteraceae and fern spores. Interpretation of Poaceae pollen was highlighted as problematic, with relatively low abundance in the cerradão (< 20%) in comparison to high abundance in lake environments (c. 30–50%). Re-examination of fossil pollen records from NKMNP revealed that modern vegetation associations were only established in the last few thousand years.     

The effect of diet on longevity,fecundity, and the sex ratio of Clitostethus arcuatus (Rossi) (Coleoptera: Coccinellidae)

Clitostethus arcuatus is a major, cosmopolitan predator of some Aleyrodidae. Field collected adult beetles were reared in the laboratory on different diets: Siphoninus phillyreae eggs, Trialeurodes vaporariorum eggs, Sitotroga cerealella eggs, or an artificial diet consisting of honey, yeast, and pollen. All experiments were conducted at 25 ± 2 °C, 65 ± 5% RH and a photoperiod of 16:8 (L:D) h. Female and male C. arcuatus consumed a mean (± SE) of 61 ± 0.6 and 27 ± 0.9 T. vaporariorum eggs d^? 1, respectively, and a mean of 56 ± 2.2 and 29 ± 1.1 S. phillyreae eggs d^? 1, respectively. Significant differences were noted between sexes and between hosts consumed by female C. arcuatus. No feeding occurred on S. cerealella eggs. Although there was a significant difference between rates of oviposition due to diet, fertility rates on different diets did not show significant differences. The sex ratio of C. arcuatus (female:male) was 51.4:48.6, 55.2:44.8, and 54.6:45:4 when adults fed on T. vaporariorum, S. phillyreae, and artificial diet, respectively. These differences were not significantly different. Average longevity (± SE) was 66.4 ± 2.6, 54.9 ± 2.5; 77.3 ± 6.9, 67.5 ± 7.2; and 86.4 ± 4.5 70.3 ± 3.6 days for female and male C. arctuatus, respectively, on T. vaporariorum, S. phillyreae and artificial diet, respectively, with significant differences between sexes and diets. Although developmental duration on T. vaporariorum was longer than ash whitefly, this difference was not significant (mean 27.68 ± 0.31 and 25.09 ± 0.21 days for predators reared on T. vaporariorum and S. phillyreae, respectively). Given its longevity and fecundity on T. vaporariorum, C. arcuatus may be a good choice for mass release on glasshouse crops infected by greenhouse whitefly.     

Development of Neoseiulus womersleyi (Schicha) and Euseius ovalis (Evans) feeding on four tetranychid mites (Acari: Phytoseiidae,Tetranychidae) and pollen

The development of the predatory mites, Neoseiulus womersleyi (Schicha) and Euseius ovalis (Evans), feeding on four tetranychid mites (Tetranychus urticae, Tetranychus kanzawai, Oligonychus mangiferus, Panonychus citri), maize pollen or Chinese loofah pollen was studied at 25 °C. Immature stages of N. womersleyi feeding on T. urticae and T. kanzawai had shorter developmental duration (4.71 and 5.02 days for females, 4.77 and 5.19 days for males, respectively) than those feeding on other food sources. Immature stages of E. ovalis females feeding on O. mangiferus and T. urticae developed in 4.99 and 5.13 days, respectively, the shortest developmental duration measured. Immature stages of E. ovalis males feeding on O. mangiferus and T. urticae developed in 5.12 and 5.37 days, respectively. The longevity of N. womersleyi males (13.31 to 14.51 days) and females (17.67 to 21.81 days) feeding on T. urticae, T. kanzawai or maize pollen was longer than the longevity of N. womersleyi feeding on O. mangiferus, P. citri or loofah pollen. E. ovalis males (12.91 to 16.74 days) and females (16.24 to 23.77 days) feeding on O. mangiferus, T. urticae or maize pollen lived longer than E. ovalis males and females feeding on T. kanzawai, P. citri or loofah pollen.     

The maternal environment determines the timing of germination in Pinus pinaster

Optimizing the germination timing is crucial for the establishment of new generations. We hypothesized that environmental maternal effects may be relevant in the fine tuning of this trait in a long-lived Mediterranean model tree. We analyzed the influence of maternal genotype, maternal environment and their interaction on the germination success and germination phenology of 8725 Pinus pinaster seeds collected from genotypes clonally replicated in two contrasting environments. Besides maternal genetic effects, the maternal environment significantly affected both the percentage and the timing of germination. Seeds from the more favourable environment germinated 7.5 days earlier and showed higher germination rate (0.93 ± 0.01 vs 0.85 ± 0.03). Seed weight significantly influenced germination time, but seed weight differences between maternal environments were not enough to explain this form of transgenerational plasticity. The effect of the maternal environment varied depending on the genotype, indicating that genetic variation in the sensitivity to the maternal environment in this pine species does exist.     

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.     

Cryopreservation of composite tissues and transplantation: Preliminary studies

Despite advances in cryobiology, the reliable cryopreservation of complex tissues has not yet been achieved. This study evaluates the viability of cryopreserved composite flaps and demonstrates the feasibility of their transplantation. Epigastric flaps were harvested from male Lewis rats. 1.5 M dimethyl sulfoxide (Me₂SO) was used as the initial cryoprotectant agent (CPA). Samples were frozen at controlled rate to −140 °C and transferred to liquid nitrogen for at least two weeks. Hematoxylin and eosin (H/E) staining, MTT tetrazolium salt assay, and factor VIII immunostaining were used to evaluate the overall histology, epithelial viability, and vascular endothelial integrity, respectively, of cryopreserved flaps. For the in vivo phase, flaps were isotransplanted to 35 recipient animals, divided into three groups: fresh (n = 10), perfused (n = 8), and cryopreserved (n = 17). Blood vessel patency was assessed via Doppler at 1, 7, and 60 days post-transplantation. For in vitro studies, cryopreserved samples (10/10) retained normal cell architecture and vascular endothelial integrity upon H/E and factor VIII staining. The viability index of cryopreserved composite flap skin (n = 10) was 11.17 ± 2.01, which was not significantly different from fresh controls (n = 10, 12.15 ± 1.32). All transplanted flaps in the fresh and perfusion groups survived with healthy color and hair growth at 60 days after operation. Survival in the cryopreserved group ranged from 2 to 60 days, with a mean of 12 days. These results demonstrate that the long term survival of cryopreserved composite tissue transplants is possible. Further studies are needed to refine protocols for the reliable cryopreservation of composite parts.     

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.     

Ecology of Salix variegata seed germination: Implications for species distribution and conservation in the Three Gorges region

The Three Gorges Project in China has brought significant changes to the region's riparian environment, posing serious challenges to biodiversity conservation. Salix variegata is a native shrub distributed on riverbanks in the Three Gorges reservoir region, and has important ecological function in the community. In this study, we studied the ecology of seed germination of S. variegata as efforts to understand its spatial distribution and develop conservation strategies. We found that soil type, soil moisture content and temperature all had significant effects on seed germination in S. variegata. Germination rate increased with the increasing soil moisture, while decreased in over-saturated soil. The germination rate in riverbank sand was higher than in sandy soil and clay loam. The optimal temperature for seed germination was 24 °C, under which 100% of germination was reached in 7 days. Seed viability and germination declined significantly with time after being released from fruits. The results suggest that soil moisture is the determining factor on seed germination, and influenced distribution of S. variegata. The tributary riverbanks above 175 m in the Three Gorges reservoir region can be used for the conservation of the species, as their fluvial patterns should have not been disturbed by the hydropower project.     

Proximate,mineral, fibre,phytate–phosphate,vitamin E,amino acid and fatty acid composition of Terminalia sericea

Terminalia sericea is widely distributed in the African Savannah bushveld. It is one of the indigenous fruit bearing trees put to multiple uses. Research has focused on the phytochemical composition of its root, bark, and leaf extracts that are used in ethnomedicine neglecting the potential of its seed. This study purposed to determine, by chemical analyses, the nutritive value of T. sericea seed. 78.8% of the seed was found to be crude protein (46.2%) and lipid (32.6%). Ash made up 6.90% of the seed mass. Linoleic and oleic acids constituted 68.63% and 14.05%, respectively, of the seed oil. Phosphorus (1121.75 ± 10.39 mg 100 g^− 1 DM) and glutamic acid (8.07 ± 0.13 g 100^− 1 DM) constituted the most concentrated mineral and amino acid, respectively. T. sericea seed could be utilized as a protein source in feeds and foods and could also be exploited as a non-conventional plant oil source of oleic acid and linoleic acid.     

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.     

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.     

Temperature requirements for dormancy break and seed germination vary greatly among 14 wetland Carex species

We evaluated dormancy loss in seeds of 14 Carex species (C. atherodes, C. brevior, C. comosa, C. cristatella, C. cryptolepis, C. granularis, C. hystericina, C. lacustris, C. pellita, C. scoparia, C. stipata, C. stricta, C. utriculata, C. vulpinoidea) under growing season and stratification conditions and determined the temperature requirements for germination. Seeds were germinated for 1 year at a diel temperature regime (5/1 °C, 14/1 °C, 22/8 °C, or 27/15 °C) or a seasonal regime (seeds moved among the four diel regimes to mimic seasonal temperatures). All species had conditionally dormant seeds at maturity. The optimal temperature for germination of most species was 27/15 °C. The 14 species were grouped by their seed viability, dormancy, and germination with a Seed Regeneration Index (SRI; range 0–1) using the results of this study and a previously published paper on stratification effects on Carex seed dormancy and germination. The eight species that had an SRI value >0.5 (C. brevior, C. comosa, C. cristatella, C. cryptolepis, C. hystericina, C. scoparia, C. stipata, C. vulpinoidea) had high seed viability (>60%) and required little to no stratification to germinate readily over a broad range of temperatures. The six species with an SRI value <0.5 (C. atherodes, C. granularis, C. lacustris, C. pellita, C. stricta, C. utriculata) generally had low seed viability (<50% and often <1%) and required stratification or particular temperatures (35/30 °C or 5/1 °C for C. stricta; 35/30 °C for C. utriculata; 27/15 °C for C. atherodes, C. lacustris, C. pellita; 5/1 °C for C. granularis) for germination ≥50%. These six species will require more attention from restoration practitioners to ensure that there are sufficient viable seeds to meet revegetation goals, that dormancy break is achieved, and that seeds are sown when temperatures are optimal for germination. The different seed germination syndromes that we found for these Carex species likely contribute to variable seed bank formation and emergence patterns, and species coexistence.     

Holdfast aggregation in relation to morphology,age, attachment and drag for the kelp Ecklonia radiata

This study quantified the prevalence of holdfast aggregation (fusion of holdfasts) for the kelp Ecklonia radiata on subtidal reefs in southwestern Australia, and tested whether morphology, age, attachment or drag were different between kelps growing alone (solitary) or in aggregates. Wave-sheltered in-shore reefs consistently had fewer aggregates than wave-exposed off-shore reefs (15–20% versus 20–30%). On average, individual thalli from aggregates were longer (97.8 cm ± 2.2 S.E. versus 88.0 cm ± 2.0 S.E.) and had smaller holdfasts (32.9 g fresh wt ± 1.7 S.E. versus 45.8 g fresh wt ± 2.9 S.E.) than solitary thalli, whereas there were no significant differences in other morphological characters, including total biomass (805.1 g fresh wt ± 38.7 S.E. versus 831.5 g fresh wt ± 38.5 S.E.), stipe length (7.93 cm ± 0.47 S.E. versus 7.65 cm ± 0.40 S.E.) and stipe diameter (12.6 mm ± 0.23 S.E. versus 13.0 mm ± 0.25 S.E.). There was no difference in age between solitary (2.7–3.0 years) and aggregated (2.4–2.8 years) individuals. While the attachment force of whole aggregates (256.5 N ± 21.6 S.E.) was found to be significantly larger than attachment force for solitary individuals (162.5 N ± 12.9 S.E.), attachment areas were also larger for aggregates (90.7 cm² ± 5.40 S.E. versus 64.3 cm² ± 5.54 S.E.) and consequently there were no differences in attachment strength between aggregates (2.92 N cm⁻² ± 0.26 S.E.) and solitary thalli (2.71 N cm⁻² ± 0.22 S.E.). Aggregates had significantly smaller (17%) roughness factors (equivalent to drag coefficients) than solitary individuals and a negative relationship (r = −0.68) between roughness factors and biomass suggested that this was related to the scope for compaction and rearrangement of the thalli. Further, there was no relationship between roughness factors of solitary individuals and the aggregates they produced when combined, suggesting that roughness factors are not additive or multiplicative. The spatial distribution of holdfast aggregates, the morphological differences between solitary and aggregated individuals as well as their attachment and drag characteristics were all consistent with aggregation reducing the rate of fatal kelp dislodgment.     

Rapid in vitro micropropagation of Agapanthus praecox

In vitro micropropagation and acclimatization for the ornamental Agapanthus praecox, are reported. The influence of different growth regulators on shoot multiplication from shoot-tip explants of A. praecox was investigated. Prolific shoot multiplication (47.3 ± 1.96 shoots per explant) was achieved on Murashige and Skoog (MS) medium supplemented with 22.2 μM benzyladenine (BA), 2.9 μM indole-3-acetic acid (IAA), and 4.5 μM thidiazuron (TDZ). Shoots were rooted on half-strength MS basal medium supplemented with 5.7 μM IAA and 2.5 μM 2-isopentenyladenine (2iP) with 11.3 ± 0.78 roots per shoot. The in vitro-raised plants were established successfully in a 1:1 (v/v) vermiculite:sand mixture when maintained in a greenhouse with 100% survival. The elongated shoots (more than 5 cm in length) were treated for rooting and acclimatization in a moistened (5.7 μM IAA and 2.5 μM 2iP) vermiculite:sand (1:1 v/v) mixture, first in the misthouse and then in the greenhouse. Rooting and acclimatization was achieved simultaneously (100%) in the misthouse which was followed by greenhouse cultivation. This system can be used for rapid mass clonal propagation of A. praecox, for conservation strategies, commercial production, gene transformation studies and to produce phytomedicines.