Campanulaceae: a family with small seeds that require light for germination

Background and Aims

The Campanulaceae is a large cosmopolitan family, but is understudied in terms of germination, and seed biology in general. Small seed mass (usually in the range 10–200 µg) is a noteworthy trait of the family, and having small seeds is commonly associated with a light requirement. Thus, the purpose of this study was to investigate the effect of light on germination in 131 taxa of the Campanulaceae family, from all five continents of its distribution.

Methods

For all taxa, seed germination was tested in light (8 or 12 h photoperiod) and continuous darkness under constant and alternating temperatures. For four taxa, the effect of light on germination was examined over a wide range of temperatures on a thermogradient plate, and the possible substitution of the light requirement by gibberellic acid and nitrate was examined in ten taxa.

Key Results

For all 131 taxa, seed germination was higher in light than in darkness for every temperature tested. Across species, the light requirement decreased significantly with increasing seed mass. For larger seeded species, germination in the dark reached higher levels under alternating than under constant temperatures. Gibberellic acid promoted germination in darkness whereas nitrates partially substituted for a light requirement only in species showing some dark germination.

Conclusions

A light requirement for germination, observed in virtually all taxa examined, constitutes a collective characteristic of the family. It is postulated that smaller seeded taxa might germinate only on the soil surface or at shallow depths, while larger seeded species might additionally germinate when buried in the soil if cued to do so by fluctuating temperatures.     

Alternating Temperatures and Rate of Seed Germination in Lentil

The effect of alternating temperatures on the times taken byseeds of lentil (Lens culinaris Medikus) to germinate was investigatedusing a two-way temperature-gradient plate. Between 5 and 25°C,warmer temperatures increased the rate of germination. Variationamong the individual seeds in the times required for germinationat different constant temperatures within this range were describedwell by a log-normal distribution of thermal times, accumulatedabove a base temperature of 1·5°C. Even with amplitudesas great as 20°C, no effect of alternation per se on thethermal time required for germination was detected—whetherthe cool temperature was applied for 8 or 16 h d^-1. Similarly,in alternating temperature regimes where the minimum temperatureof the diurnal cycle was between 0°C and the base temperature,the thermal times required for germination (where no thermaltime accrued during the periods when temperature was below T_b)were in close agreement with those values provided by the modeldetermined at warmer constant temperatures. However, where theminimum temperature applied was < 0°C the germinationof all but the earliest germinators was delayed beyond modelpredictions, and more so where the sub-zero minimum temperaturewas applied for 16 rather than 8 h d^-1. The results, therefore,contradict the view that alternation in temperature per se reducesthe thermal time required for seed germination. Rather, rateof germination responds instantaneously to current temperature,but prolonged exposure to sub-zero temperatures can result indamage sufficient to delay germination when seeds are returnedto regimes warmer than the base temperature.Copyright 1994,1999 Academic Press Lens culinaris Medikus, lentil, seed germination, alternating temperatures, thermal time, temperature-gradient plate     

An Investigation of the Influence of Constant and Alternating Temperature on the Germination of Cassava Seed using a Two-dimensional Temperature Gradient Plate

The germination of cassava seed in response to various constantand alternating temperature regimes within the range 19–40°C was investigated using a two-dimensional temperaturegradient plate. It was found that almost all seeds were incapableof germination unless the temperature for part of the day exceeded30 °C and the mean temperature was at least 24 °C. However,dormant seeds required environments where the temperature forpart of the day exceeded 36 °C, the mean temperature wasat least 33 °C, and the amplitude of the diurnal temperaturealteration was within the range 3–18 °C. Providingthese conditions were met, the times spent at the upper andlower temperatures within a diurnal cycle were not critical.Hermetic storage of the seed for 77 days at 40 °C with 7.9per cent moisture content did not influence the pattern of germinationin response to constant and alternating temperatures. It issuggested that an alternating temperature regime of 30 °Cfor 8 h/38 °C for 16 h applied for a minimum of 21 daysis appropriate for cassava seed viability tests. Manihot esculenta Crantz, cassava, germination, dormancy, temperature     

Germination of Stored Cassava Seed at Constant and Alternating Temperatures

Cassava seed is only capable of germinating over a restrictedrange of constant temperatures. During storage the optimum constanttemperature for germination decreases from about 35 to 30 °Cor possibly less. The rate at which the optimum temperaturechanges during dry storage increases with increase in storagetemperature over the range 0 to 40 °C. Some alternating-temperatureregimes (16 h at the lower temperature; 8 h at the higher temperature)can provide conditions as favourable for germination as theoptimum constant temperatures. Furthermore, it has been shownthat temperature alternation itself is stimulatory because whenthe range of the alternation does not include the optimum constanttemperature value, percentage germination is often higher thancould be obtained at any constant temperature within the range,though this stimulatory response declines during storage. Forthese reasons it is provisionally recommended that cassava seedshould be germinated at 25/35 °C which is as stimulatorya treatment as any which has so far been investigated and hasthe advantage of encompassing the range over which the optimumconstant temperature changes during storage. Manihot esculenta Crantz, cassava, germination, dormancy, seed viability, storage of seeds, after-ripening     

Hormonal and temperature regulation of seed dormancy and germination in <Emphasis Type="Italic">Leymus chinensis</Emphasis>

Fluctuating temperature plays a critical role in determining the timing of seed germination in many plant species. However, the physiological and biochemical mechanisms underlying such a response have been paid little attention. The present study investigated the effect of plant growth regulators and cold stratification in regulating Leymus chinensis seed germination and dormancy response to temperature. Results showed that seed germination was less than 2 % at all constant temperatures while fluctuating temperature significantly increased germination percentage. The highest germination was 71 % at 20/30 °C. Removal of the embryo enclosing material of L. chinensis seed germinated to 74 %, and replaced the requirement for fluctuating temperature to germinate, by increasing embryo growth potential. Applications of GA₄₊₇ significantly increased seed germination at constant temperature. Also, inhibition of GA biosynthesis significantly decreased seed germination at fluctuating temperatures depending upon paclobutrazol concentration. This implied GA was necessary for non-dormant seed germination and played an important role in regulating seed germination response to temperature. Inhibition of ABA biosynthesis during imbibition completely released seed dormancy at 20/30 °C, but showed no effect on seed germination at constant temperature, suggesting ABA biosynthesis was important for seed dormancy maintenance but may not involve in seed germination response to temperature. Cold stratification with water or GA₃ induced seed into secondary dormancy, but this effect was reversed by exogenous FL, suggesting ABA biosynthesis during cold stratification was involved in secondary dormancy. Also, cold stratification with FL entirely replaced the requirement of fluctuating temperature for germination with seeds having 73 % germination at constant temperature. This appears to be attributed to inhibition of ABA biosynthesis and an increase of GA biosynthesis during cold stratification, leading to an increased embryo growth potential. We suggest that fluctuating temperature promotes seed germination by increasing embryo growth potential, mainly attributed to GA biosynthesis during imbibitions. ABA is important for seed dormancy maintenance and induction but showed less effect on non-dormant seed germination response to temperature.     

FACTORS AFFECTING GERMINATION IN GREENHOUSE-PRODUCED SEEDS OF OXALIS CORNICULATA,A PERENNIAL WEED

A study was conducted to investigate the physiological responses of greenhouse-produced Oxalis corniculata seeds to light, temperature, moist heat treatment, aging, and season of production. Fresh seeds exhibited over 90% germination and required low levels of light (5 μmol m^-2 s^-1, 400–700 nm) to germinate. Seeds germinated over a broad, yet seasonally-dependent range of incubation temperatures. Seeds produced in winter had the narrowest temperature range of germination (15 to 25 C) and the lowest germination percent (44% at 2 wk) at optimum temperature (17 C); seeds produced in summer had the widest temperature range of germination (10 to 30 C) and the highest germination percent (93% at 2 wk) at optimum temperature (17 C). Incubation at non-optimum temperatures between 5 and 40 C suppressed or slowed the rate of germination until seeds were placed at optimum temperature, where full germination subsequently occurred. Moist heat treatment at temperatures over 40 C resulted in varying degrees of inhibition of subsequent germination. When seeds were stored dry in laboratory conditions, three of four seed lots examined retained over 80% germination capacity until ca. 8 months; 50% capacity remained after ca. 15 months. These results indicate that the seasonal temperature and daylength effects on maternal plants in the greenhouse environment are major determinants of seed germination characteristics of O. corniculata.     

Seed germination of endemic species from Kerguelen phytogeographic zone

 Seeds of six endemic species and two circumpolar species were collected from several sites on Kerguelen and tested for germination between 5 and 25°C on thermogradient bars. Pringlea antiscorbutica and Colobanthus kerguelensis had a near 100% viability with an initial germination temperature of ca. 24°C. Colobanthus attained nearly 100% viability from 5 to 25°C whereas Pringlea germination declined markedly below 20°C. In Poa kerguelensis and P. cookii germination was poor and varied between sites. The three Ranunculus spp. showed various degrees of dormancy. In Lyallia kerguelensis, although the seed appeared mature, a variety of treatments could only partly release an apparently deep dormancy. These data are discussed with respect to field observations on reproduction from seed. Received: 1 February 1996/Accepted; 28 April 1996     

Germination responses to temperature responsible for the seedling emergence seasonality ofPrimula sieboldii E. Morren in its natural habitat

Temperature requirements for the breaking of seed dormancy and germination inPrimula sieboldii E. Morren and the annual surface-soil temperature regime in one of its natural habitats were investigated in order to clarify the germination responses determining the seedling emergence seasonality of the species. In a grassland nature reserve in an abandoned flood plain of the Arakawa River, natural seedling emergence of the species was shown to be restricted to mid- to late-spring before the closure of seasonal vegetational gaps, when the daily mean soil surface temperature reached about 15°C, accompanied by large daily fluctuations of about 10°C. Mature seeds collected in late June were never able to germinate at any constant temperature in the range of 8–40°C unless they had been previously subjected to moist-chilling treatment. The proportion of seeds which were released from dormancy increased with increasing duration of the moist-chilling treatment at 2°C, 70–85% of seeds becoming germinable at 16–28°C after 12 weeks of pretreatment at 2°C. The thermal time required for the germination of the thus-pretreated seed population was 905–1690 Kh with a base temperature of around 5°C. Fluctuating temperatures between 24°C and 16 or 12°C had a remarkable dormancy-breaking effect, inducing considerably quick germination in most of the seeds previously subjected to 2°C moist-chilling for 8 weeks.     

Germination Responses of Taraxacum platycarpum Seeds to Temperature

Abstract Germination responses of Taraxacum platycarpum seeds to temperature were examined under laboratory conditions to investigate the emergence-season choice mechanism of the seeds. Almost all the newly collected seeds were non-dormant. Under constant temperature conditions, maximum percentage germination (approximately 90%) was attained at temperatures 6–16°C, where simple linear relationships were observed between the temperature and the rates of germination, i.e. the reciprocals of the time taken to germinate by seed subpopulations with 10–80% germination. Thermal time required for germination of the subpopulations ranged from 600 Kh (degree Kelvin × hours) to 1500 Kh with a relatively constant base temperature of about 2.5°C. Lower limit temperature for germination was slightly below 6°C. Higher limit temperature for germination has the normal distribution with the mean ±SD of 19±2.5°C. Pre-exposure of imbibed seeds to temperatures higher than the higher limit temperature for germination, 25 and 30°C, had no effect on the germinability and the rate of germination at a circa-optimum temperature. Moist chilling treatment at 4°C caused an increase in the variation of germination rate within the seed population, but no evidence for dormancy-inducing or breaking effects was obtained.     

Asymbiotic seed germination and in vitro seedling development of <Emphasis Type="Italic">Cyrtopodium punctatum</Emphasis>: a propagation protocol for an endangered Florida native orchid

Cyrtopodium punctatum Lindley is an endangered epiphytic orchid restricted in the United States to southern Florida. Due to its ornamental value, the species was extensively collected from the wild during the past 100 years. Today, only a few plants remain in protected areas. As part of a conservation plan for the species, procedures for asymbiotic seed germination were developed. Five asymbiotic orchid seed germination media (PhytoTechnology Orchid Seed Sowing Medium, Knudson C, Malmgren Modified Terrestrial Orchid Medium, Vacin &; Went Modified Orchid Medium, and ½-strengh Murashige &; Skoog) were examined for their effectiveness in promoting seed germination and protocorm development under a 16/8 h L/D photoperiod and dark (0/24 h L/D). The influence of photoperiod on growth and development was also examined. Seeds were germinated under a 16/8 h, 12/12 h, 8/16 h L/D photoperiod, at 25 ± 3°C and allowed to develop in vitro for 10 weeks. After 10 weeks, developing seedlings were transferred to Sigma Phytatrays and returned to their assigned photoperiod treatments for continued seedling development for an additional 15 weeks. Highest germination occurred in 0/24 h L/D on PhytoTechnology Orchid Seed Sowing Medium and seedlings displayed more advanced development when cultured under 16/8 h L/D photoperiod after 15 weeks in Phytatrays. Thirty-five week old seedlings potted in coconut husk growing medium exhibited 90% survival following 5 weeks acclimatization to greenhouse conditions. This asymbiotic seed germination protocol for C. punctatum will facilitate future reintroduction projects involving this endangered species.     

Effects of different nitrogen compounds and temperatures on the germination ofAvena sterilis spp.macrocarpa Mo.

The effect of several nitrogen compounds on the germination of the first seed of each spikelet ofAvena sterilis spp.macrocarpa Mo. was studied on Petri dishes placed in germination chamber. The compounds assayed were ammonium sulphate, ammonium nitrate, urea and calcium nitrate, generally used as fertilizer for cereal crops, as well as a control supplied with water only The concentration of the assayed compounds were 100, 1000, 2500 and 5000 ppm either at a constant temperature of 15 ° C or at alternated temperatures 5°C and 15 °C, for 16 and 8 h, respectively     

野豌豆属4种植物种子萌发的积温模型分析

以青藏高原野豌豆属窄叶野豌豆(Vicia angustifolia)、山野豌豆(V. amoena)、歪头菜(V. unijuga) 3种野生植物与一种当地栽培植物救荒野豌豆(箭筈豌豆) (V. sativa) ‘兰箭3号’种子为材料, 在5、10、15、20、25及30 ℃下进行萌发实验, 应用种子萌发的积温模型对上述4种植物萌发对温度的响应特征进行了比较分析。结果表明: 1)基于萌发速率(1/T_g)对种子萌发温度最低温T_b值的估计受萌发率(g)的影响较小; 与此不同, 除‘兰箭3号’种子外, 对萌发最高温T_c值的估计, 受到g的显著影响。 这表明种群内所有种子个体萌发的T_b值相对恒定, 但T_c值在有些物种中变异较大; 2)基于重复概率单位回归分析估计的种子萌发T_b值与基于萌发速率估计的值较为接近; 而由此方法估计的T_c值则与萌发率为50%时的估计值较为接近; 3)相比多年生豆科植物歪头菜和山野豌豆, 一年生豆科植物箭筈豌豆‘兰箭3号’与窄叶野豌豆具有相对较低的T_b与T_c值; 4)积温模型可准确地预测休眠破除后豆科植物种子在不同温度条件下的萌发进程。     

Effect of temperature and photoperiod on the induction of diapause in the mite Metaseiulus occidentalis

Diapause in a predaceous mite, Metaseiulus occidentalis, from a Californian vineyard population is a photoperiodically induced, facultative, adult reproductive diapause in females. The laboratory-determined critical photophase at 19°C was estimated at 11·2 hr. At 16°C, the critical photophase under laboratory conditions was approximately 11·6 hr. Temperature influenced the photoresponse of M. occidentalis so that diapause was entirely averted at temperatures of 22, 25, and 30°C. Aestival diapause at higher temperatures and long photophases was lacking. Development was continuous under constant darkness at all the temperatures tested. Diapause termination in laboratory-reared mites occurred spontaneously under the inductive conditions. Under constant 19°C temperatures, females responded to photophases so that diapause was terminated most rapidly under a 16 hr photophase (in 18·6 days); the 12 and 8 hr photophases, at this temperature, were next in their effectiveness, with 27·9 and 73·0 days, respectively, required for termination.     

SEED GERMINATION STUDIES IN MUSA. II. ALTERNATING TEMPERATURE REQUIREMENT FOR THE GERMINATION OF MUSA BALBISIANA

Stotzky , G., and Elsie A. Cox . (Central Research Labs., United Fruit Co., Norwood, Mass.) Seed germination studies in Musa. II. Alternating temperature requirement for the germination of Musa balbisiana. Amer. Jour. Bot. 49(7): 763–770. Illus. 1962.—Alternating temperatures were found to be required for the germination of seeds of Musa balbisiana. The temperature differentials optimal for germination in soil are dependent upon both the high and low temperatures, and range from 8–23 C. Germination is maximal when the seeds are held 6–12 hr at the high (27–35 C) and 12–18 hr at the low (12–18 C) temperatures. Some germination can be induced by short exposures to alternating temperatures followed by constant high temperatures, but continuous exposure to alternating temperatures is necessary for maximum germination. Excised embryos develop better at constant than at alternating temperatures, showing that the mechanisms affected by alternating temperatures reside elsewhere in the seed. Alternating temperatures are also required for germination of mechanically scarified seeds, although the temperature differentials are less than those necessary for intact seeds, indicating that the action of alternating temperatures is not on the permeability of the integuments.     

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.     

Development of a Threshold Model to Predict Germination of Populus tomentosa Seeds after Harvest and Storage under Ambient Condition

Effects of temperature, storage time and their combination on germination of aspen (Populus tomentosa) seeds were investigated. Aspen seeds were germinated at 5 to 30°C at 5°C intervals after storage for a period of time under 28°C and 75% relative humidity. The effect of temperature on aspen seed germination could not be effectively described by the thermal time (TT) model, which underestimated the germination rate at 5°C and poorly predicted the time courses of germination at 10, 20, 25 and 30°C. A modified TT model (MTT) which assumed a two-phased linear relationship between germination rate and temperature was more accurate in predicting the germination rate and percentage and had a higher likelihood of being correct than the TT model. The maximum lifetime threshold (MLT) model accurately described the effect of storage time on seed germination across all the germination temperatures. An aging thermal time (ATT) model combining both the TT and MLT models was developed to describe the effect of both temperature and storage time on seed germination. When the ATT model was applied to germination data across all the temperatures and storage times, it produced a relatively poor fit. Adjusting the ATT model to separately fit germination data at low and high temperatures in the suboptimal range increased the models accuracy for predicting seed germination. Both the MLT and ATT models indicate that germination of aspen seeds have distinct physiological responses to temperature within a suboptimal range.     

Characterizing Ipomopsis rubra (Polemoniaceae) germination under various thermal scenarios with non-parametric and semi-parametric statistical methods

Time-to-event analysis represents a collection of relatively new, flexible, and robust statistical techniques for investigating the incidence and timing of transitions from one discrete condition to another. Plant biology is replete with examples of such transitions occurring from the cellular to population levels. However, application of these statistical methods has been rare in botanical research. Here, we demonstrate the use of non- and semi-parametric time-to-event and categorical data analyses to address questions regarding seed to seedling transitions of Ipomopsis rubra propagules exposed to various doses of constant or simulated seasonal diel temperatures. Seeds were capable of germinating rapidly to >90 % at 15–25 or 22/11–29/19 °C. Optimum temperatures for germination occurred at 25 or 29/19 °C. Germination was inhibited and seed viability decreased at temperatures ≥30 or 33/24 °C. Kaplan–Meier estimates of survivor functions indicated highly significant differences in temporal germination patterns for seeds exposed to fluctuating or constant temperatures. Extended Cox regression models specified an inverse relationship between temperature and the hazard of germination. Moreover, temperature and the temperature × day interaction had significant effects on germination response. Comparisons to reference temperatures and linear contrasts suggest that summer temperatures (33/24 °C) play a significant role in differential germination responses. Similarly, simple and complex comparisons revealed that the effects of elevated temperatures predominate in terms of components of seed viability. In summary, the application of non- and semi-parametric analyses provides appropriate, powerful data analysis procedures to address various topics in seed biology and more widespread use is encouraged.     

Effect of temperature and cold stratification on seed germination of the Mediterranean wild aromatic Clinopodium sandalioticum (Lamiaceae)

A germination study was carried out on seeds of Clinopodium sandalioticum (Bacch. & Brullo) Bacch. & Brullo ex Peruzzi & Conti (Lamiaceae), a wild aromatic plant endemic to Sardinia. Seeds were incubated at a range of constant (5–25°C) and an alternating temperatures regime (25/10°C), with 12 hours of irradiance per day. The results achieved at 10°C were also compared with those obtained after a period of cold stratification at 5°C for three months. Final seed germination ranged from ca. 28% (5°C) to ca. 72% (25/10°C). A base temperature for germination (T_b) of ca. 5°C and a thermal constant for 50% germination (S) of 89.3°Cd were identified and an optimal temperature for germination (T_o) was estimated to be comprised between 20 and 25°C. Cold stratification negatively affected seed viability and germination at 10°C. Although a typical “Mediterranean germination syndrome”, could not be detected for C. sandalioticum seeds, these results were coherent with those previously reported for other Mediterranean Lamiaceae species.