Tomato pollen development: stages sensitive to chilling and a natural environment for the selection of resistant genotypes

Abstract The time during which pollen development is most sensitive to chilling was investigated. Five cultivars of tomato (Lycopersicon esculentum Mill.) bearing flower buds at different stages of development were kept at 7°C for 1 week under 12-h light periods, during which time growth stopped. After returning the plants to minimum temperatures of 18°C, the presence of chromatin in the pollen was assessed daily as the flowers reached anthesis. The results suggested that there are two stages of acute sensitivity to cold during pollen development, each of which results in cold-stressed plants having pollen empty of chromatin. The first and most sensitive stage is about 11.2 d (SE = 0.3 d) before anthesis, and this is followed by a second stage of sensitivity about 5.6±0.2 d before anthesis. Flowers that had wholly developed under simulated natural temperatures that decreased diurnally from a maximum of 18°C to a minimum of 7°C also had defective pollen, but pollen of normal appearance was regained within 14°d on return to higher temperatures. Plants of L. esculentum, and a form (LA 1363) of the wild species L. hirsutum from high altitudes in the Andes, as well as F1 and F3 generations of their hybrid, were grown to the flowering stage at an altitude of 600 m in Hawaii and then grown for a further 30°d at 2000 m, where night temperature was below 10°C. The high altitude environment severely affected the quality of pollen produced and its release from the stamen in L. esculentum, but not in L. hirsutum LA 1363. The results with the hybrids suggested that such tropical mountain environments can be used as a natural phytotron in the selection of chilling resistance that is only expressed in the mature plant.     

Reproductive ecology of dioecious Siparuna (Monimiaceae) in Ecuador–a case of gall midge pollination

The reproductive ecology of the dioecious genus Siparuna , belonging to the primitive family Monimiaceae, was investigated in Ecuador. Thirteen species were studied, and species growing at high altitudes (above 500 m) were compared with species growing at lower altitudes. The genus has specialized flowers pollinated by Cecidomyiidae (Asynapta sp. and other genera) that lay eggs chiefly in male flowers since these are more easily accessible than female flowers due to their morphology. Several traits ensure that pollen transfer is accomplished: 1. long flowering of individuals and single flowers; 2. specific floral morphology (unisexuality, landing platform, restricted pore) allowing the flowers to be visited by Cecidomyiidae seeking egg-laying sites, and 3. a low number of ovules that have to be pollinated in each flower. The low number of ovules per flower results in a high pollen-ovule (P/O) ratio even though pollen production in male flowers is only moderate.
High altitude species had larger flowers, higher pollen-ovule ratios, fewer ovules per female flower, and longer periods of anthesis. Gender differences were seen in the longer functionality of female flowers; the larger numbers of male flowers produced; and the average diameter at breast height (DBH) biased for one of the two sexes depending on the species investigated, with no clear trend in the size distributions of individuals of both sexes. Sex ratios departed significantly from unity both to male and female bias but usually were in favour of males. There were no general differences in the ratio of functional male to female flowers at the time of observation nor in the sizes of male and female flowers. The evolution of unisexual flowers in Siparuna can be explained as a result of the differential predation by larvae: unimportant in male flowers, destructive if occurring in female flowers.     

Altitudinal variations in germination and growth responses ofReynoutria japonica populations on Mt Fuji to a controlled thermal environment

The authors examined altitudinal variations in the thermal responses of seed germination and seedling growth inReynoutria japonica (=Polygonum cuspidatum) under controlled environmental conditions. Seed populations were collected from different altitudes on Mt Fuji in Japan. The mean seed weight of the upland populations (above 1500 m) was significantly (1.5-fold) heavier than that of the lowland populations (below 1400 m). Under the lowest temperature regime of 15/10°C (day/night) the upland populations showed a significantly higher percentage and speed of germination than the lowland populations; this was not significant under higher temperature regimes. These results indicate that the germination traits of the upland populations on Mt Fuji are favorable for colonization in their cold habitats (low temperature and short growing season). Growth and shoot development were compared between the seedlings grown from seeds collected at altitudes of 700 and 2420 m. The upland seedlings showed a significantly larger biomass and leaf area than the lowland seedlings at 15°C, but there was no difference at 25°C. The difference in biomass at 15°C was attributed to the difference in seed weight. The upland seedlings produced a significantly larger number of branches with smaller and more numerous leaves at both 15°C and 25°C. these developmental traits of the upland seedlings were considered to represent the adaptation of the life form to upland environments. It was concluded that theR. japonica populations along an altitudinal gradient on Mt Fuji can be classified into two ecotypes, whose distribution border lies at an altitude of about 1400–1500m. In this study, the seed weight and germination traits of twoR. japonica seed populations collected in Chiba Prefecture were briefly compared with those of the lowland populations on Mt Fuji.     

Can floral consumption by fish shape traits of seagrass flowers?

Seagrasses are marine flowering plants with hydrophilous pollination. This abiotic pollination by water assumes absence of flower-animal interaction, but animals can interfere in this process through consumption of reproductive structures. We studied predation on male flowers by fish for three dioecious seagrass species (Thalassia testudinum, Syringodium filiforme and Halodule wrightii) in the Mexican Caribbean. Seagrass flowers have a highly reduced or absent corolla and florivores directly consumed the anthers with pollen. The foliar structures (tepals, bracts or sheaths) protecting the male flower buds were removed by hand in situ. The floral buds were followed by videos or taking pictures at regular intervals and most (56–100 %, depending on seagrass species and experimental setting) artificially denuded male flower buds were consumed within 24 h by juvenile fish of various species. Histochemical analysis showed that the pollen and embedding mucilage were rich in polysaccharides and proteins, thus potentially nutritious. The seagrasses had copious production of pollen (between 0.2 and 1.2 × 10⁶ pollen per flower, depending on the species). But T. testudinum and S. filiforme were often pollen limited, and the probability of fruit set was reduced ~50 % when the females were at the distance of 1 and 5–6 m from the males flowers, respectively. Under natural conditions, depredation on pre-anthesis male flowers in the three species was low because flower bud emergence (few hours) and pollen release (1–4 h) were ephemeral processes. In addition, the release of pollen of T. testudinum occurred at dusk when herbivorous fish became inactive. These life-cycle characteristics aid to avoid excessive pollen consumption by fish, however, whether they are anti-predator strategies or mere adaptations for submarine pollination remains to be established.     

Cumulative temperatures for prediction of the beginning of ash (Fraxinus excelsior L.) pollen season

The aim of this study is the analysis of the meteorological conditions leading to the release of ash pollen in order to predict the beginning of the pollen season in Zurich, Switzerland. For 12 years over a period of 14 consecutive years, it appears that ash pollen release begins when the cumulated mean daily temperatures starting on January 1 reach 220.7°C, admitting a variation of ±30°C that more or less corresponds to 3 consecutive days at 10°C.     

Effectiveness of nectarivorous birds and honeybees as pollinators of Banksia spinulosa (Proteaceae)

Abstract The effectiveness of nectarivorous birds and honeybees (Apis mellifera) as pollinators of Banksia spinulosa (Proteaceae) was investigated. Birds visited inflorescences in the early, mid and late flowering seasons. In contrast, honeybees visited only on days in the late flowering period when maximum temperatures exceeded 15°C. Self pollen remained on pollen presenters of flowers for up to 5 days in the early and mid periods. In the late period, when honeybees visited inflorescences, self pollen was removed within 2 days. Pollen removal was similar for caged (birds excluded) and open inflorescences in the late period, indicating that most pollen was removed by honeybees. In the early and mid periods, honeyeaters pollinated 22% and 27% of flowers on open inflorescences, respectively. In the late period, when both birds and bees visited inflorescences, 64–73% of flowers on open inflorescences were pollinated. Foraging by honeybees resulted in pollen deposition as 38% of flowers on caged inflorescences were pollinated. Throughout the flowering season a similar number of pollen grains was deposited per stigma. There were 3.0–3.7 pollen grains per stigma on open inflorescences in the late period, although only 2.0 grains per stigma on caged inflorescences. In the early and mid periods, fewer caged than open inflorescences produced fruits, indicating the importance of honeyeaters to reproductive success at these times. In contrast, in the late period when honeybees visited inflorescences, fruit-set was similar on caged and open inflorescences. Overall, these results indicate that honeybees were effective pollinators of B. spinulosa.     

Seed dormancy-breaking and germination requirements ofDrosera anglica,an insectivorous species of the Northern Hemisphere

Seeds of Drosera anglica collected in Sweden were dormant at maturity in late summer, and dormancy break occurred during cold stratification. Stratified seeds required light for germination, but light had to be given after temperatures were high enough to be favorable for germination. Seeds stratified in darkness at 5/1 °C and incubated in light at 12/12 h daily temperature regimes of 15/6, 20/10 and 25/15 °C germinated slower and to a significantly lower percentage at each temperature regime than those stratified in light and incubated in light. Length of the stratification period required before seeds would germinate to high percentages depended on (1) whether seeds were in light or in darkness during stratification and during the subsequent incubation period, and (2) the temperature regime during incubation. Seeds collected in 1999 germinated to 4, 24 and 92 % in light at 15/6, 20/10 and 25/15 °C, respectively, after 2 weeks of stratification in light. Seeds stratified in light for 18 weeks and incubated in light at 15/6, 20/10 and 25/15 °C germinated to 87, 95 and 100 %, respectively, while those stratified in darkness for 18 weeks and incubated in light germinated to 6, 82 and 91 %, respectively. Seeds collected from the same site in 1998 and 1999, stratified in light at 5/1 °C and incubated in light at 15/6 °C germinated to 22 and 87 %, respectively, indicating year-to-year variation in degree of dormancy. As dormancy break occurred, the minimum temperature for germination decreased. Thus, seed dormancy is broken in nature by cold stratification during winter, and by spring, seeds are capable of germinating at low habitat temperatures, if they are exposed to light.     

Genotypic influence on in vitro induction, dormancy length, advancing age and agronomical performance of potato microtubers (Solanum tuberosum L.)

Microtubers of 13 cultivars, largely grown in Italy and other European countries, were induced. They were stored in the dark at 3°C for different periods (28, 56, 84 and 105 days), prior to being transferred to 20°C for between 4 and 17 weeks. Following removal to room temperature, sprouting was recorded and dormancy duration quantified. Dormancy decreased from 28.1 to 19.9, 11.1 and 7.8 days with reduced time of storage. Cvs Arsy, Nicola and Jaerla took consistently more time for dormancy release. The dormancy duration was linearly and inversely correlated with the length of storage. After sprouting, tubers were held at 20°C for various intervals and a range of physiological ages (0, 368, 720 and 1008 degree days) were accumulated. The field comparison of microtubers evidenced a plant growth response and tuber yield/plant affected by the cultivar and physiological age. In early cultivars (Jaerla), a better performance was shown by younger tubers; the opposite trend was noted in Alpha (a later cultivar) with an increase in stems/plant, tubers/plant and tuber yield/plant for tubers with greater physiological age. Like conventional seed tubers, microtubers showed differences in optimum physiological age associated with cultivar earliness. This study has provided some indications on how to enhance emergence and haulm development of plants from microtubers.     

Environmental analysis of airborne pollen occurrence,pollen source distribution and phenology of <Emphasis Type="Italic">Fraxinus angustifolia</Emphasis>

Narrow-leafed ash (Fraxinus angustifolia) is a common polygamous tree growing on the banks of rivers in the western Mediterranean region. Pollination occurs during winter, and the tree’s pollen is among the most abundant during that season. This work aims to relate the phenology of pollen shedding, source tree distribution, meteorology and airborne pollen occurrence for the species. Aerobiological sampling was conducted in Badajoz (south-western Spain) using a Hirst volumetric sampler over 24 years (1993–2016). Trees were geo-localized in a circle 500 m in diameter surrounding the pollen sampler. During the last two periods, pollination phenology was studied in 10 specimens, five in the surroundings of the pollen station and five 3 km apart, at a frequency of 3–4 days on average. Moreover, a detailed analysis of pollen occurrence was conducted for these two periods. Daily data for the whole period and hourly data over the last 2 years were used, including pollen monitoring and meteorology. A comparison was made between pollen occurrence and source distribution. The main pollen season lasted on average 53 (28–75) days. Average values were less than 10 grains m^?3, except for two periods of 23–24 grains m^?3. Daily data and hourly data correlation with meteorology showed different signs in correlation analysis. Hourly analysis showed that the maximum concentration occurred just after noon. Most pollen was recorded at an average temperature of 9 °C. Analysis of pollen sources and pollen occurrence showed a close relationship between predominant wind directions and tree distribution. Peaks of phenology were not coincident with pollen peaks. No trends in pollination were found. Non-homogeneous distribution of pollen sources for Fraxinus angustifolia provided a suitable tool to demonstrate that wind direction plays a relevant role when aerobiological data are interpreted according to source distribution. A limitation in phenology analysis and aerobiological data was noted in the narrow-leafed ash species.     

Low temperature induced floral abortion in chickpea: relationship to abscisic acid and cryoprotectants in reproductive organs

Cold stress (temperature <8 °C) during reproductive growth of chickpea is detrimental to flowering and pod set. Cold-sensitive genotypes either delay their flowering or abort their flowers produced during intermittent warm spells upon exposure to cold stress. To find out the mechanism of floral abortion in chickpea, a cold-sensitive genotype (PBG1) was grown under controlled (28 °C/12 °C) and cold-stressed conditions (12–15 °C/4–6 °C). Pod set was 95% in controlled conditions versus 20% under cold stress. The relative leaf water content (RLWC) of stressed plants was decreased by 26% over controls. The retained and aborted flowers harvested from the plants growing under cold stress along with flowers from control plants were examined for functioning of gametes, endogenous levels of ABA, sucrose, glucose, fructose, trehalose and proline in whole flowers, and their male (pollen, anthers) as well as female (stigma–style, ovary) parts. Flowers from stressed plants showed a decrease in functioning of gametes in comparison to those from control plants. The pollen germination (in vivo and in vitro) and viability did not differ significantly between the retained and aborted flowers of the stressed plants but stigma receptivity and ovary viability were significantly lower in aborted flowers. The ABA content in aborted flowers was 23% higher than in retained flowers. The content of sucrose, glucose and fructose in aborted flowers was 40, 25 and 23% lower, respectively, and the content of trehalose and proline was 50 and 73% lower, respectively, than in flowers on the plant. Male parts of aborted flowers contained 7–12% greater concentration of ABA and solutes than those retained on the mother plant but the female parts of the former, especially ovary contained 25% more ABA and 23–26% lower solute levels. Since the male parts of aborted flowers were relatively little affected by cold stress, the events in female parts appeared to be primarily responsible for floral abortion in chickpea. Reciprocal pollination between plants growing in controlled and stressed conditions indicated greater impairment of female parts than male parts under cold stress. Proline (5 mM) application to stressed plants improved the floral retention and pod set by 33%.     

Physiological dormancy in seeds of tropical montane woody species in Hawai`i

Worldwide, there is relatively little information on seed dormancy and germination of tropical montane species. Our aim was to help fill this knowledge gap by conducting seed dormancy/germination studies on woody species from this vegetation zone in Hawai`i. All species had water-permeable seeds with a fully developed embryo. Seeds of 29 species (23 genera) were incubated in light/dark at 15/6, 20/10 and 25/15°C and germination monitored at 2-week intervals for 16–128 weeks. Seeds of Chenopodium oahuense, Dubautia menziesii and Silene lanceolata were non-dormant (ND) and those of 26 other species had physiological dormancy (PD); 10 of the 26 species had conditional PD. The optimum germination temperature regime(s) was (were) 25/15°C, 17 species; 25/10 and 20/10°C, 2; 20/10°C, 6; 20/10 and 15/6°C, 2; and 15/6°C, 2. Worldwide, PD in the woody genera included in our study is more common than ND. In addition to its contribution to the world biogeography of seed dormancy/germination, this study will be useful to conservation biologists who need to germinate seeds of tropical montane species.     

Physiological factors limit fruit set of tomato (Lycopersicon esculentum Mill.) under chronic, mild heat stress

The effects of chronic, mild heat stress on fruit set, fruit production, release of pollen grains, photosynthesis, night respiration and anther dehiscence were examined in tomatoes (Lycopersicon esculentum Mill.) differing in high‐temperature sensitivity. Plants were grown under three temperature regimes: (1) 28/22 or 26/22 °C (optimal temperature); (2) 32/26 °C (high temperature); and (3) 32/26 °C day/night temperatures relieved at 28/22 °C for 10 d before anthesis, then returned to 32/26 °C (relieving treatment). FLA 7156 was the only cultivar with fruit set at 32/26 °C. All five cultivars, however, had fruit set under the relieving treatment (RT). The longer the relief, the higher the percentage of fruit set. Longer periods of relief also increased the number of pollen grains released, and linear regression analysis showed a significant relationship between the number of pollen grains released and the percentage of fruit set. Germination of pollen grains was also lowered in high‐temperature‐grown plants. The number of pollen grains produced, photosynthesis and night respiration did not limit fruit set under chronic, mild heat stress, however. This suggested that cultivar differences in pollen release and germination under heat stress are the most important factors determining their ability to set fruit.     

The development of Puccinia hordei on barley cv. Zephyr

Germination of uredospores of Puccinia hordei was similar on cover-slips and on the first leaves of barley seedlings (cv. Zephyr) at 100 % r.h. over the range 5–25 °C, being greatest at 20 °C. At 15, 20 and 25 °C maximum germination was attained in 6 h. No uredospores germinated on coverslips in humidities below saturation. The numbers of pustules which subsequently developed on plants incubated at 5, 10, 15 or 18 °C and 100 % r.h. for varying periods up to 24 h, were directly related to rise in temperature and length of incubation. The time from inoculation to eruption of pustules (generation time) was 6 days at 25 °C, 8 days at 20 °C, 10 days at 15 °C, 15 days at 10 °C and 60 days at 5 °C. Pustule production on inoculated plants which had been kept at 5 °C was rapidly accelerated when they were transferred to 20 °C. Data obtained at constant temperatures were used to predict generation times of the fungus in the field. The productivity of pustules, determined as weight of uredospores, was examined at 10, 15 and 20 °C. Significantly more spores were produced at 15 than at 10 °C and most were produced at 20 °C. The results are discussed in relation to those obtained by other workers and to the development of brown rust in the field.     

Chilling and heat requirements for the prediction of the beginning of the pollen season of Alnus glutinosa (L.) Gaertner in Ponferrada (León, Spain)

Winter-flowering trees such as the alder (Alnus glutinosa (L.) Gaertner) can survive periods of adverse climatic conditions, entering a period of dormancy in the early fall. The end of dormancy and the start of the pollen season require a period of low temperatures followed by another of warm temperatures. These requirements were studied from 1995 to 2002, in order to develop a model to predict the onset of the alder pollen season in Ponferrada (Spain). Chilling accumulation took place from late October to late December or early January. The best result was obtained with a threshold temperature of 6.5 °C and an average of 848 chilling hours (CH). Heat requirements were calculated at maximum temperature, an average 143 growth degree days (GDD) were needed, with a threshold temperature of 0 °C. In order to validate models, predicted values were compared with real values for 2002–2003, 2003–2004 and 2004–2005, years not used in developing the models. Predictions for the pollen-season start-date differed only slightly from observed dates: in 2002–2003 predicted and observed dates were the same, in 2003–2004 there was a difference of 7 days and in 2004–2005 a difference of 3 days.     

Seed dormancy and germination in Dodonaea viscosa (Sapindaceae) from south-western Saudi Arabia

Dodonaea viscosa (Sapindaceae) is widespread in the mountainous highlands of the southwestern part of Kingdom of Saudi Arabia, where it is a medicinally important species for the people in Saudi Arabia. Seeds of this species were collected from Mount Atharb in Al-Baha region, at an altitude of 2100 m. The aims of this study were to determine if the seeds of D. viscosa have physical dormancy (i.e. a water-impermeable seed coat) and, if so, what treatments would break dormancy, and what conditions promote germination after dormancy has been broken. The dormancy-breaking treatments included: soaking of seeds in concentrated sulfuric acid (H₂SO₄) for 10 min, immersion in boiling water for 10 min and exposure to 50 °C for 1 min. After seeds had been pre-treated with H₂SO₄, to break dormancy, they were incubated at constant temperatures from 5 to 35 °C, under 12-h photoperiods or in continuous darkness, and germination recorded. Salinity tolerance was investigated by incubating acid-scarified seeds in different concentrations of mM NaCl in the light at 25 °C.Untreated seeds had low final germination 30%. Seeds that had been acid-scarified, immersed in boiling water or exposed to 50 °C all achieved 91% subsequently when incubated at 25 °C. Thus, seeds of this species in Saudi Arabia have physical dormancy, which can be broken by all three treatments designed to increase the permeability of the testa. After pre-treatment, there was a broad optimum constant temperature for germination that ranged between 5 and 25 °C but germination was inhibited by higher temperatures (30 and 35 °C). Light had little effect on this germination response. Scarified seeds were also sensitive to salinity, with the highest germination in distilled water and complete inhibition in 400 mM NaCl. Seeds that failed to germinate in saline treatments were mostly able to germinate on transfer to distilled water, suggesting osmotic inhibition.     

Morphophysiological dormancy in seeds of the ANA grade angiosperm Schisandra arisanensis (Schisandraceae)

We determined the kind of seed dormancy in Schisandra arisanensis, an ANA grade ([A]mborellales [N]ymphaeales [A]ustrobaileyales) angiosperm with medicinal value. Seeds have small underdeveloped embryos, and following seed maturity their length increased approximately 360% before radicle emergence. Germination was delayed 6–8 weeks, and the percentage and rate were much higher at 15/6, 20/10 and 25/15°C than at 30/20°C. For seeds incubated at 5/5°C (8 weeks) → 15/6°C (4 weeks) → 20/10°C (8 weeks) → 25/15°C (12 weeks) → 20/10°C (5 weeks), embryos grew at 15/6°C → 20/10°C, and almost all seeds that germinated (89%) did so at 20/10°C → 25/15°C. When seeds were incubated in a complementary temperature sequence, 25/15°C (12 weeks) → 20/10°C (8 weeks) → 15/6°C (4 weeks) → 5/5°C (9 weeks) → 15/6°C (4 weeks), embryos grew at 25/15°C → 20/10°C. Nearly all seeds that germinated (93%) did so at 25/15°C → 20/10°C and at 15/6°C following 9 weeks at 5/5°C. Based on the temperature requirements for embryo growth and seed germination, seeds of this species have non‐deep simple morphophysiological dormancy (C_1bB).     

Culture And Staining of Pollen Grains of Ricinus communis

Germinating and growing pollen grains (male gametophytes) of Ricinus communis L. in liquid culture is achieved as follows: Pollen is collected over a 10-15 min period from mature anther clusters which have been removed from the male flowers and which have been kept at 25° C and 40-60% relative humidity. Samples weighing between 2.5 and 5.0 mg are brought as quickly as possible into a Desicote treated vial containing 17% sucrose and 30 ppm H₃BO₃ in boiled distilled water. The proportion (w/v) of pollen to culture solution should be 1:100. Shed pollen is kept in a humidity chamber whenever it is not being handled. The air in the culture vial is replaced by O₂ at the pressure of 1 atmosphere plus 5 lb and the sealed vials are shaken gently for 8-10 hr while partially immersed in a waterbath kept at 30° C. The pollen is fixed by the addition to the incubation suspension of an absolute alcohol-lactic acid (4:1) fixing fluid. The proportion used is 36 parts of fixing fluid to 1 part of culture solution. The fixed pollen can be stored in the fixative. Smears are prepared by applying single drops of the constantly agitated suspension of fixed pollen to a microscope slide. After each drop has spread out and dried, an additional drop is added until 10-20 have been applied. The preparations are stained by adding a drop of 1% acetic-orcein and are sealed with fingernail lacquer. The method is well adapted to the following types of studies: pollen germination, physiology of pollen tube growth, morphology of the male gametocyte, and physiology and cytology of the generative cell and nucleus.     

Pollen germination and pollen tube growth in Fraxinus pennsylvanica

With regard to adaptation of green ash (Fraxinus pennsylvanica Marshall) to ecological conditions in Croatia, pollen germination and pollen tube length after 2, 4 and 6 hours were examined in vitro at 10, 15, 20 and 25°C during two years 2001 and 2002. Narrow leaved ash (F. angustifolia Vahl) pollen served as a control in 2002. The year, time and temperature, and the interaction between time and temperature were significant for both germination percentage and pollen tube length. Interactions year × temperature and year × time were significant for pollen tube length only. The highest germination percentage (17.86% in 2001 and 19.40% in 2002) of green ash pollen was at 15°C after 6 hours. The pollen tube length was greatest at 20°C (393.46 μm) in 2001 and 25°C (899.50 μm) in 2002 after 6 hours. Narrow leaved ash pollen had the highest germination percentage (19.22%) at 20°C after 6 hours and was significantly reduced at 25°C. The pollen tube length was greatest at 25°C (518.90 μm) after 6 hours. It can be concluded that green ash pollen has satisfactory germination in ecological conditions in Croatia and that the optimum temperature for pollen germination is higher than 20°C.     

Effects of endogenous ABA levels and temperature on cedar (Cedrus libani Loudon) bud dormancy in vitro

Axillary and apical buds of in-vitro-propagated cuttings of Cedrus libani are unable to burst at 24 °C, but this inhibition was overcome at 30 °C. Here we have used cedar microcuttings to investigate whether the levels of endogenous hormones vary with bud dormancy and temperature. We analysed the levels of abscisic acid, indole-3-acetic acid, zeatin, isopentenyladenine and their major metabolites using HPLC purification and fractionation of the samples coupled to an ELISA method for hormonal quantitation involving several antibodies elicited against each hormonal family. Abscisic acid levels in microcuttings with dormant buds were higher than those in microcuttings with growing buds. At 24 °C, needles accumulated more abscisic acid than at 30 °C. In addition, when needles were removed, but growth release was achieved at 24 °C. Abscisic acid supplied at 30 °C induced the formation of dormant buds. These results suggest that abscisic acid accumulation in the needles can explain the bud dormancy of cedar microcuttings at 24 °C. Received: 14 November 1997 / Revision received: 16 January 1998 / Accepted: 5 May 1998     

Releasing primary dormancy in <Emphasis Type="Italic">Avena fatua</Emphasis> L. caryopses by smoke-derived butenolide

The effect of smoke and smoke-derived butenolide in releasing dormancy of caryopses (referred to as seeds) of the economically important weed Avena fatua L. was studied. Seeds of A. fatua are dormant after harvest. Both smoke-water and butenolide, applied continuously, removed dormancy in darkness at 15, 20 and 25°C and slightly at 30°C. Butenolide was very active at a concentration of 10⁻⁸ M. Butenolide at 10⁻⁸ M was also able to remove dormancy at 20°C when applied for 12 or 24 h at 4°C or for 3 to 24 h at 20°C. Sensitivity to butenolide decreased with longer preincubation times in water. This compound was less effective in releasing dormancy in the light than in darkness. Dormancy release by butenolide involves induction of cell-cycle activity just before coleorhiza protrusion. Stimulatory effects of smoke-water and butenolide were also observed in respect of seedling growth and vigor.