Comparative study of allozyme polymorphism in groups of pine trees (Pinus sylvestris L.) with different seed productivity

Genotypes of 196 Pinus sylvestris L. plants from 10 natural populations of five Ukrainian regions have been determined using 19 polymorphic isozyme loci. Variability of quantity of full-grained, empty-grained and underdeveloped seeds in the cones of these plants has been studied. The basic indexes of genetic polymorphism were determined for 6 samples presented by 18-19 trees with high and low productivity of the full-grained, empty-grained and underdeveloped seeds. The maximum amount of rare alleles and genotypes as well as the highest heterozygosity (Ho = 0.285) were typical for the sample of plants with the maximum quantity of empty-grained seeds in the cones.     

Systemic spread of downy mildew in basil plants and detection of the pathogen in seed and plant samples

Downy mildew on sweet basil (Ocimum basilicum L.) occurs worldwide. Contaminated seeds are considered as the primary inoculum source. So far no strategy to control the disease is available. Hence, the use of pathogen-free seeds is the only alternative to prevent disease outbreaks. Therefore, a rapid diagnostic method for seed testing is urgently needed. The sensitivity of a specific PCR method for direct detection of the downy mildew pathogen Peronospora belbahrii on basil samples, particularly on seeds, was evaluated. The applied PCR method proved to be very sensitive for direct detection of the pathogen on seeds and plant samples. The PCR detection limit of P. belbahrii in artificially infested seeds corresponded to the DNA amount of a single spore per seed. Additionally, the systemic spread of the pathogen from naturally infected seeds was investigated. The experiments showed that outgrowing basil plants were latently infected with the downy mildew pathogen, and the infection continued within the plant. Contaminated seeds were harvested from symptomless latently infected plants. These results support the implementation of PCR-based detection in a seed certification scheme and the necessity to control the pathogen on seeds. The PCR method can also be used for evaluation of pathogen control on seeds based on detection of the pathogen in outgrowing plants.     

低温保存技术在顽拗性种子种质保存中的利用

由于顽拗性种子不耐脱水且对低温敏感,常规保存方法难以达到长期保存的目的。因此,(超)低温保存顽拗性种子种质是最理想的方法。顽拗性种子的低温保存,应用较多的是玻璃化法和两步法。诸多因素影响着低温保存的成败,如种子或胚的含水量水平、溶液低温保护剂效应、降温冰冻与解冻方式、水合过程以及后培养等,这些需深入探索与解决。除顽拗性种子脱水耐性和低温敏感性机理外,植物细胞的冻害和抗冻机理也亟需探明,以便找到最佳冷冻方法,制定长期保存种质基因的最佳方案。     

Endozoochorous seed dispersal by free-ranging herbivores in an abandoned landscape

Seed dispersal is a crucial process for the dynamics and maintenance of plant populations. Free-ranging animals are effective dispersal vectors because they can move between similar habitats and transport seeds into favourable environments. Dung samples from two species of common free-ranging mammals—deer and wild boar—were used to study endozoochorous dispersal of seeds in a military training area in western Bohemia. The area was abandoned after WWII, and the military training area was established in 1953. The vegetation consists of shrublands and dry grasslands. Data on the local species pool of grassland herbs and forbs were collected to compare the characteristics of dispersed versus non-dispersed plants. Deer and wild boar dispersed 84 plant species; however, species composition of seedlings emerging from dung samples showed significant differences between dispersal vectors and notable change across the growing season. 80% of all seedlings extracted from the dung samples belonged to stinging nettle, Urtica dioica. From trait analyses, seeds of endozoochorous plants had a higher longevity index in the soil seed bank than non-endozoochorous plants and more often possessed a mucilaginous surface. Our results show that deer and boar are successful, though not substitutable dispersers.     

Detection and seed transmission of Bermudagrass phytoplasma in maize in Turkey

During 2017, maize cultivation areas in the provinces of Adana and Kahramanmara? (Turkey) were surveyed to inspect maize plants with symptoms similar to those associated with of phytoplasma disease, that is, yellowing, short internodes and small corncobs. Thirty fields were inspected and two hundred samples from symptomatic and asymptomatic plants were collected, together with insects considered as potential vectors of phytoplasmas. All samples were assayed by polymerase chain reaction (PCR) and subsequently analysed by restriction fragment length polymorphism and sequencing to identify the phytoplasmas detected in the plant material and insects. Results of laboratory assays and phylogenetic analyses showed that the Bermudagrass white leaf phytoplasma ('Candidatus Phytoplasma cynodontis') was present in both maize plants and seeds, showing 99% sequence identity with other reported phytoplasma strains from GenBank, whereas no PCR amplifications were obtained from tested insects. The seeds of infected plants, sown in an insect‐proof screenhouse, produced plantlets that were found PCR‐positive for the Bermudagrass white leaf phytoplasma, indicating its seed transmission.     

Co-evolution of seed size and seed predation

Using the evolutionarily stable strategy (ESS) approach in a model for the co-evolution of seed size and seed predation, I show that seed size variation within individual plants is favoured if there is a trade-off in the predator's attack rate for different seed sizes. A single seed size is not evolutionarily stable because a predator that is optimally adapted to one particular seed size cannot prevent invasion by plants with a different seed size. The model generates the following predictions. The ESS consists of a continuous range of seed sizes. Small seeds tend to be attacked more frequently than big seeds. Plants with many resources and plants with low (frequency-independent) juvenile mortality have more variable seeds than plants with few resources and a high juvenile mortality. Seed size variation is higher in fluctuating populations regulated by seed predation alone than in stable populations (partially) regulated by seedling competition. Predator searching behaviour does not directly affect the ESS seed size range, but may have an indirect effect by affecting population stability or the significance of seedling competition as a population regulating mechanism. Moreover, seed size distributions are found to be more skewed in favour of small seeds if predation is spatially non-uniform than if predation is more even. Application of the model to systems of several co-evolving plant and predator species is discussed.     

Seed vigor and plant competitiveness resulting from seeds of Eupatorium adenophorum in a persistent soil seed bank

Seeds in a persistent soil seed bank (PSSB) provide an effective way to maintain plant population and community stability. Seeds that persist in soil incur physiological costs of maintaining viability and vigor, thus, the growth capability of resulting plants may be reduced. However, a lot of functional roles of the PSSB have been deduced from seed germination capability, and little consideration has been given to interspecific and intraspecific competitive ability of the resulting plants. Eupatorium adenophorum was used as the study species to compare germination of different artificially aged PSSB seeds and competition at different densities between resulting plants of aged and freshly produced seeds. Seed burial caused decreases in survival rates but not germination speed. During the 175-day growth period, the individual biomass, average height, basal stem diameter and leaf number of plants from aged PSSB seeds were little lower than that of plants germinated from freshly produced seeds. However, the differences were not significant at any densities. Thus, (1) although seeds stored in soil exhibited a very high death rate, they maintained a high vigor for germination, and (2) resulting plants from PSSB seeds exhibited good competiveness to plants from new seeds of the same population. The results further confirm the significance of PSSB in maintaining stability of plant populations and communities.     

动物与植物种子更新的关系Ⅱ．动物对种子的捕食、扩散、贮藏及与幼苗建成的关系

植物的繁殖体总是面临来自各类生物（如昆虫、脊椎动物、真菌）的捕食风险。因动物捕食引起的种子死亡率影响植物的适合度、种群动态、群落结构和物种多样性的保持。种子被捕食的时间和强度成为植物生活史中发芽速度、地下种子库等特征的主要选择压力,而种子大小、生境类型等因素也影响动物对植物种子的捕食。捕食者饱和现象被认为是植物和种子捕食者之间的高度协同进化作用的结果,是限制动物破坏种子、提高被扩散种子存活率的一种选择压力。大部分群落中的大多数植物种子被动物扩散。种子扩散影响种子密度、种子被捕食率、病原体攻击率、种子与母树的距离、种子到达的生境类型以及建成的植株将与何种植物竞争,从而影响种子和幼苗的存活,最终影响母树及后代植物的适合度。种子被动物扩散后的分布一般遵循负指数分布曲线,大多数种子并没有扩散到离母树很远的地方。捕食风险、生境类型、植被盖度均影响动物对种子的扩散。植物结实的季节和果实损耗的过程也体现了其对扩散机会的适应。许多动物有贮藏植物种子的行为。动物贮藏植物繁殖体的行为,一方面调节食物的时空分布,提高了贮食动物在食物缺乏期的生存概率;另一方面也为种子萌发提供了适宜条件,促进了植物的扩散。于是,植物与贮食动物形成了一种协同进化关系,这种关系可能是自然界互惠关系（mutualism)的一种。影响幼苗存活和建成的因子包括种子贮蒇点的微生境、湿度、坡向、坡度、林冠盖度等。许多果食性动物吃掉果肉后,再将完好的种子反刍或排泄出来。种子经动物消化道处理后,发芽率常有所提高。     

Experimental determination of seed emergence and carry‐over in the soil seed bank of the herbaceous perennial,Trachymene incisa (Apiaceae)

Abstract Many populations of herbaceous perennial plants contain seeds stored in a soil seed bank. The contribution of seeds to population persistence is an important parameter in population models but germination rates of known‐age seeds are difficult to obtain because individual seeds cannot easily be followed. Although Trachymene incisa Rudge plants produce copious seeds that are dispersed into the soil, the existence of a seed bank has not been confirmed. To quantify the potential for a seed bank fresh seeds of T. incisa were sown into experimental seed banks in the eucalypt‐dominated Agnes Banks Woodland in western Sydney, NSW. A recent fire provided the opportunity to compare germination in the burnt and unburnt vegetation. Density of seed sowing and time of maturation/dispersal of seeds were manipulated in 75 seed cages. Emergence of seeds after 5 months was significantly higher for the earliest planting date but after 1 year, germination of seeds planted in the later weeks increased, and the final germination for all weeks was 28%. Density of sowing and the recent fire did not affect emergence. A second experiment planted over a broader time span (9 weeks instead of 3 weeks) confirmed the effect of planting date but also found significant spatial variation on a scale of tens of metres. Laboratory germination rates of over 70% confirmed that the seeds were viable and non‐dormant when sown in the field cages. The carry‐over of non‐germinated seed in the soil seed bank is estimated to be about 70% after 2 years, implying that a cohort of seeds would not be depleted through germination alone for up to 40 years. The potential for a long‐lived seed bank in this species is interesting because the plants are also capable of resprouting from their rootstock after fire, giving them characteristics of both resprouters and seeders.     

Masting,seed dispersal and seed predation in the cycad Macrozamia communis

Summary Little is known about the adaptive value of mast seeding, a common phenomenon in temperate trees and shrubs. Masting is likely to affect both seed dispersal and seed predation. In systems where similar taxa of animals are involved in these two processes, the consequences of mast seeding are likely to be particularly complicated. This study examined the effects of mast seeding in a cycad, Macrozamia communis, on the dispersal of seeds, the pattern of dispersion of seeds and post-dispersal predation on seeds. Dispersal of seeds by possums was poorer from source plants in a masting population than from source plants in an adjacent, non-masting population. This resulted in fewer seeds per seeding female plant in the masting plot being dispersed to favourable sites. We conclude that this is caused by the feeding behaviour and movements of possums in the masting site. The abundance of seeds in this site did not satiate the post-dispersal predators, native rats. In fact, more seeds in this site were eaten than in the nonmasting site. We suggest that the mast seeding observed in M. communis may not be adaptive, but is more likely a consequence of other factors which synchromize flowering within local populations.     

The role of salinity in seed maturation of the euhalophyte Suaeda salsa

Controlled conditions were used to investigate how salinity maintains the salt tolerance of seeds and seedlings of the euhalophyte Suaeda salsa. Seeds were harvested from S. salsa plants that had been treated with 1 or 500 mM NaCl for 113 days in a glasshouse. The results showed that high salinity (500 mM NaCl) increased chlorophyll concentration and oxygen production in embryos of maturing seeds. At 500 mM NaCl, the phosphatidylglycerol and sulfoquinovosyldiacylglycerol levels and the digalactosyldiacylglycerol/monogalactosyldiacylglycerol ratio were higher in young seedlings derived from seeds whose source plants were cultured in 500 mM rather than in 1 mM NaCl. When seeds were incubated with 600 mM NaCl, the conductivity and malondialdehyde concentration in the embryos was greater if the source plants had been cultured in 1 mM rather than in 500 mM NaCl. The opposite pattern was evident for seedling survival and shoot weight. In conclusion, salinity during seed maturation may increase the salt tolerance of seeds and seedlings by increasing the oxygen production in the embryos of the maturing seeds and by changing the lipid composition of membranes in the seedlings.     

Enhanced seed defenses potentially relax selection by seed predators against serotiny in lodgepole pine

Serotiny, the retention of seeds in a canopy seed bank until high temperatures cause seeds to be released, is an important life history trait for many woody plants in fire‐prone habitats. Serotiny provides a competitive advantage after fire but increases vulnerability to predispersal seed predation, due to the seeds being retained in clusters in predictable locations for extended periods. This creates opposing selection pressures. Serotiny is favored in areas of high fire frequency, but is selected against by predispersal seed predators. However, predation also selects for cone traits associated with seed defense that could reduce predation on serotinous cones and thereby relax selection against serotiny. This helps explain the elevated defenses in highly serotinous species. However, whether such interactions drive variation in seed defenses within variably serotinous populations has been studied rarely. We investigated the effects of phenotypic selection exerted by red squirrel (Tamiasciurus hudsonicus) predation on Rocky Mountain lodgepole pine (Pinus contorta latifolia) seeds. Squirrels preferentially harvested cones with more and larger seeds, indicating a preference for a higher food reward. We found evidence for stronger selection on trees with serotinous cones, which presumably accounts for the elevated defenses of and lower predation on serotinous compared to non‐serotinous cones. Lower levels of predation on serotinous cones in turn lessen selection against serotiny by squirrels. This has important implications because the frequency of serotiny in lodgepole pine has profound consequences for post‐fire communities and ecosystems widespread in the Rocky Mountains.     

The role of Japanese macaques (Macaca fuscata) as endozoochorous seed dispersers on Kinkazan Island, northern Japan

We studied the characteristics of seeds within faeces, an important aspect of endozoochorous seed dispersal, in Japanese macaques Macaca fuscata inhabiting Kinkazan Island, northern Japan. We intermittently collected faecal samples from 1999 to 2009 (N = 1294) and examined the rate of seed occurrence, species/life-form composition, number of seeds, and their intact rate. Seeds were found within faecal samples during every month, but their characteristics changed monthly: the rate of seed occurrence and the number of plant species within faecal samples were greater in summer and fall, and the intact rate and number of intact seeds observed within single faecal samples were also higher during these seasons than spring and winter. These results suggest that Japanese macaques on Kinkazan act as seed dispersers in summer and fall and that they disperse intact seeds into wider areas within the forest through defecation. During the study period, we observed seeds from a total of 35 plant species from 22 families in our samples. In addition to those of woody plants, we also observed seeds from as many as 12 herbaceous plants, for which sympatric sika deer (Cervus nippon) have historically been considered the sole seed dispersal agents. The intact rate of seeds was significantly negatively correlated with the seeds’ mean cubic diameter, and this relationship was strengthened for non-fleshy fruits. We also conducted regional comparisons of the characteristics of defecated seeds in order to address whether regional variations in the diet of the macaques affect their efficacy as seed dispersers, both in terms of quantity and quality. Macaques living in the natural habitats of Kinkazan and Yakushima dispersed more seeds of tall tree species than do macaques inhabiting the human-altered areas of in Shimokita and Kashima. The number of plant species represented within single faecal samples also varied geographically, being greater in Yakushima. This pattern likely resulted from Yakushima's warm temperate climate, as the other three study sites occur in the cool temperate region. Our results suggest that the composition of seeds dispersed by Japanese macaques is not rigid, but is determined instead by the vegetation found in a given habitat.     

Aerial seed bank in a cold desert annual-ephemeral species: Role of anatomical structure of stem and delayed fruit dehiscence in timing of seed dispersal

Why the fruits are retained on dead upright herbaceous plants and how this relates to seed dispersal and timing of germination remain unclear. Stems of the annual Euclidium syriacum (Brassicaceae) with infructescences bearing indehiscent silicles remain upright after plants die in the spring. We investigated the effect of anatomical structures of stem and pedicle and delayed silicle dehiscence on seed dispersal phenology of this species. For comparison, sections were made of the stem of the annual Goldbachia laevigata (Brassicaceae), which has stems that fall over when plants die. Compared to G. laevigata, the stem of E. syriacum has vascular bundles that are closer together, a thicker xylem and phloem, more fibers, a thicker perimedullary zone and a smaller pith diameter:stem diameter ratio. The thickened pedicle did not form an abcission layer. By late October, 5–20% of seeds were dispersed, depending on the position of infructescences on the plant. Snow covered the plants in late autumn and when it melted in mid-April many of the plants had fallen over, with a high number of seeds germinating in attached silicles; seedlings became rooted in soil. After snowmelt, 14–15% of the silicles on the remaining upright plants contained seeds; all seeds were dispersed by early July. The anatomical structures of the stem and pedicle plus the delayed dehiscence of silicles explain the presence of an aerial seed bank in E. syriacum and delay of germination of many of seeds until spring. Further, pieces of upright plants are broken off and dispersed by wind, which helps to explain the wide distribution of E. syriacum in the cold desert.     

First report of seed dispersal by ants in Dicentra peregrina (Papaveraceae), an alpine plant in the Japanese Alps

Seed dispersal by ants, known as myrmecochory, is commonly observed among various plant taxa. The seeds of these plants have an elaiosome to attract ants. In Japan, myrmecochory has been well studied in several lowland plant species, but not in highland plant species that grow above the tree line. We investigated whether the seeds of Dicentra peregrina, known as the “queen of the alpine plants” in Japan, are carried by Formica gagatoides ants at 2510 m a.s.l. on Mt Norikura, Kita‐Alps, Japan. We observed F. gagatoides workers picking up D. peregrina seeds by grasping the elaiosome and carrying the seeds into their nests. We inferred from the observed ant behavior and the seed morphology that D. peregrina is a myrmecochorous species.     

Dynamics of storage reserve deposition during Brassica rapa L. pollen and seed development in microgravity

Pollen and seeds share a developmental sequence characterized by intense metabolic activity during reserve deposition before drying to a cryptobiotic form. Neither pollen nor seed development has been well studied in the absence of gravity, despite the importance of these structures in supporting future long-duration manned habitation away from Earth. Using immature seeds (3-15 d postpollination) of Brassica rapa L. cv. Astroplants produced on the STS-87 flight of the space shuttle Columbia, we compared the progress of storage reserve deposition in cotyledon cells during early stages of seed development. Brassica pollen development was studied in flowers produced on plants grown entirely in microgravity on the Mir space station and fixed while on orbit. Cytochemical localization of storage reserves showed differences in starch accumulation between spaceflight and ground control plants in interior layers of the developing seed coat as early as 9 d after pollination. At this age, the embryo is in the cotyledon elongation stage, and there are numerous starch grains in the cotyledon cells in both flight and ground control seeds. In the spaceflight seeds, starch was retained after this stage, while starch grains decreased in size in the ground control seeds. Large and well-developed protein bodies were observed in cotyledon cells of ground control seeds at 15 d postpollination, but their development was delayed in the seeds produced during spaceflight. Like the developing cotyledonary tissues, cells of the anther wall and filaments from the spaceflight plants contained numerous large starch grains, while these were rarely seen in the ground controls. The tapetum remained swollen and persisted to a later developmental stage in the spaceflight plants than in the ground controls, even though most pollen grains appeared normal. These developmental markers indicate that Brassica seeds and pollen produced in microgravity were physiologically younger than those produced in 1 g. We hypothesize that microgravity limits mixing of the gaseous microenvironments inside the closed tissues and that the resulting gas composition surrounding the seeds and pollen retards their development.     

Photocontrol of seed germination in the hemiparasite Buchnera hispida (Scrophulariaceae)

Abstract Buchnera hispida, a facultative root parasite of grasses and graminaceous crops, has a light requirement for germination. Studies were carried out on the effects of varying photoperiods with or without preceding dark incubation, on seed germination. Buchnera seeds showed long-day behaviour, since they germinated at all photoperiods including continuous light, and longer photoperiods were more effective in triggering seed germination than shorter photoperiods. Also, effects of red and far-red light indicated that the phytochrome system is operative in the light-induced germination of Buchnera. Although dark incubation in water before illumination was not absolutely necessary for germination, it caused the seeds to respond more rapidly to light. The longer the time of the dark incubation the more responsive the seeds were to photoperiod except when 15 min light was given. The effectiveness of a preceding dark incubation in making Buchnera seeds sensitive to rapid light action was completely inhibited at 4°C. This is in agreement with the hypothesis that a reaction partner of the far-red absorbing form of phytochrome is produced during dark incubation of Buchnera seeds. Such an intermediate has also been reported in some positively photoblastic seeds of non-parasitic flowering plants.     

Avena fatua L. seed and seedling nutrient dynamics as influenced by mycorrhizal infection of the maternal generation

Abstract. The objective of this study was to determine how mycorrhizal infection of one generation of plants influences the nutrient dynamics of seeds and seedlings comprising the subsequent generation. We showed that, for Avena fatua L., seeds produced by mycorrhizal (M) plants consistently contained significantly more phosphorus (particularly the phytate P and residual P fractions) than seeds produced by non-mycorrhizal (NM) plants. We also followed the development of spikelets produced by M and NM plants. The rates of increase in spikelet dry weight and nitrogen content were largely unaffected by mycorrhizal infection. However, the rate of P accumulation into spikelets was significantly increased by mycorrhizal infection. Greater endosperm P reserves in seeds produced by M plants were associated with greater rates of P accumulation in resultant seedlings. Moreover, offspring plants (all NM) produced by M mother plants had significantly higher root and rhizosphere phosphatase, ATPase and phytase activities than offspring plants produced by NM mother plants. This persistent maternal effect has never before been described. Our results suggest that mycorrhizal infection of one generation of plants may have substantial positive effects on the offspring generation, and thus, may influence plant population dynamics.     

Nitrate, a signal relieving seed dormancy in Arabidopsis

Nitrate is an important nitrogen source for plants, but also a signal molecule that controls various aspects of plant development. In the present study the role of nitrate on seed dormancy in Arabidopsis was investigated. The effects of either mutations affecting the Arabidopsis nitrate reductase genes or of different nitrate regimes of mother plants on the dormancy of the seeds produced were analysed. Altogether, data show that conditions favouring nitrate accumulation in mother plants and in seeds lead to a lower dormancy of seeds with little other morphological or biochemical differences. Analysis of germination during seed development indicated that nitrate does not prevent the onset of dormancy but rather its maintenance. The effect of an exogenous supply of nitrate on seed germination was tested: nitrate in contrast to glutamine or potassium chloride clearly stimulated the germination of dormant seeds. Data show, moreover, that the Arabidopsis dual affinity nitrate transporter NRT1.1 (CHL1) may be involved in conveying the nitrate signal into seeds. Thus, nitrate provided exogenously or by mother plants to the produced seeds, acts as a signal molecule favouring germination in Arabidopsis. This signalling may involve interaction with the abscisic acid or gibberellin pathway.