Effect of seed age,stratification, and soaking on germination of wild asparagus (Asparagus acutifolius L.)

Wild asparagus (Asparagus acutifolius L.) is a widespread species found in all the Mediterranean areas. The spears are highly valued by consumers and owing to its frugality, this species is a feasible new crop with high income potential, especially for Mediterranean marginal areas. Currently, the cultivation of this species is limited because of its low and erratic seed germination that makes difficult the production of seedlings for plant propagation. In this research, non-after-ripened (1 month-old) and after-ripened seeds (dry stored at room temperature for 13 months) were exposed for 30 days in the dark to three moist stratification treatments: cold (5 °C), warm (23 °C) or no stratification; subsequently they were soaked for 12 h in warm water (35 °C) or not soaked. The effect of these pre-germination treatments on three germination parameters (germination percentage, time to 50% of final germination – T₅₀ – and germination pattern) was studied, as well as some possible seed dormancy forms involved therein. The 1-year dry storage period proved to be effective in after-ripened seeds by enhancing seed sensitivity to the subsequent pre-germination treatments. After-ripened seeds exhibited higher and more rapid germination compared to non-after-ripened seeds. Soaking, cold or warm moist stratification had similar single effect on non-after-ripened seeds (27% germination). With after-ripened seeds, only soaking or warm stratification were effective (47% germination) when singularly applied, while cold stratification did not improve germination. By combining stratification and soaking treatments, a higher germination for both non-after-ripened and after-ripened seed-lots was achieved. The highest germination was obtained when after-ripened seeds were stratified and soaked (76%), without any significant difference between cold or warm stratification. Single or combined application of moist stratification (regardless of the temperature used) and soaking resulted always in a faster germination compared to that of no-treated seeds and especially with after-ripened seeds (T₅₀ = 6 days). A non-deep type 1 physiological dormancy can be hypothesized for the seeds of this species. Low stratification temperature induce secondary dormancy in after-ripened seeds that can be removed by soaking them at 35 °C for 12 h.     

Levels of physiological dormancy and methods for improving seed germination of four rose species

Low seed germination is a major problem in commercial rose propagation and breeding and is species-dependent. The present work selected four rose species previously un-examined to explore effective methods for improving seed germination and the relevant dormancy mechanism and its levels in seven experiments. The results showed that both pulp and achenes from the four rose shrubs had chemical substances that significantly inhibited seed germination with the inhibitory effect was more pronounced in pulp extract than of achenes. Single treatments of H₂SO₄ scarification, short-term cold stratification (<16 weeks) or warm stratification were less effective in breaking dormancy as indicated by lower germination index than their combinations. Comprehensive comparisons showed that among the six treatments the most effective for breaking dormancy was H₂SO₄ scarification followed by warm plus cold stratification, then H₂SO₄ scarification followed by cold stratification and finally warm plus cold stratification. Scarification with H₂SO₄ for 2–4 h ordinal followed by warm stratification at 20 °C for 4 weeks and cold stratification at 5 °C for 8 weeks was the best pretreatment for increasing seed germination percentage for Rosa multibracteata (81.4 ± 2.9%), Rosa hugonis (13.1 ± 6.0%), and Rosa filipes (62.7 ± 5.7%); and H₂SO₄ scarification for 4 h followed by cold stratification at 5 °C for 12 weeks was the best pretreatment for Rosa sericea (46.7 ± 8.7%). Our results suggest that these four species have only physiological dormancy caused by integrative roles of pulp, pericarp and embryo. The level of physiological dormancy was ranked as R. hugonis > R. sericea > R. filipes > R. multibracteata.     

Interspecific variations in seed germination of Corylopsis

This study was initiated to investigate the differences in germination percentages and rates between Corylopsis coreana Uyeki and Corylopsis sinensis var. calvescens Rehder & E.H. Wilson following a warm stratification (WS) and cold stratification (CS), and to study the effect of different WS temperatures interacting with different durations of CS. Warm stratification at 10 °C, 15 °C, 20 °C, and 25 °C was given for 1 month (1 M 10 °C, 15 °C, 20 °C, and 25 °C WS) followed by 0 M, 1 M, 2 M, and 3 M of CS at 5 °C (0 M, 1 M, 2 M, 3 M CS) and seeds were germinated in an air conditioned greenhouse maintained at 18.5 °C/18 °C. On average, less than 1% of C. coreana seeds germinated when sown without any WS and CS or with 1 M 15 °C, 20 °C, and 25 °C WS without CS treatment. However, 26% C. coreana seeds germinated after 1 M 10 °C WS without any CS treatment. Germination was not affected by WS temperatures when followed by 2 M 5 °C CS. It is concluded that C. coreana exhibited low seed germination at 10 °C and that this temperature could be considered the upper limit of CS for C. coreana. Only 2 M CS was required for more than 90% seeds to germinate. However, C. sinensis var. calvescens required longer than 3 M CS for more than 29% seeds to germinate. This clearly shows that there is an interspecific variation in optimum dormancy-breaking requirements.     

Effects of after-ripening,stratification and GA3 on dormancy release and on germination of wild asparagus (Asparagus acutifolius L.) seeds

Seeds of wild asparagus (Asparagus acutifolius L.) were treated and compared in this research to investigate seed dormancy class and level involved in this species. Four seed lots were compared: (i) freshly harvested seeds in 2007 (07Fr); (ii) freshly harvested seeds in 2008 (08Fr); (iii) after-ripened (AR) 2007 seeds dry stored in glass jars (ARg); (iv) AR 2007 seeds dry stored in paper bags (ARp). The 07Fr seeds were exposed to (1) chemical scarification combined with gibberellic acid (GA₃) levels (0, 200, 400, and 600 mg L⁻¹) and to (2) 28-day moist stratification at 5 and 23 °C, and two sequences of 5/23 °C combined with 0 and 400 GA₃ mg L⁻¹ levels, and (3) together to the 08Fr and AR seeds were exposed to 56-day moist stratification at 5, 23, or 5/23 °C. With the 08Fr and AR seed lots this last stratification treatment was combined with 0 or 800 GA₃ mg L⁻¹ levels. The dormancy depth of 08Fr (32% germination) was less than 07Fr seeds (2%). The latter after-ripened during dry storage and when stored in glass germinated more (47.5%) than in paper (12%). Stratification for 4 weeks was ineffective in improving germination of 07Fr seeds; when chemically scarified they did not germinate at all. The highest (nearly 70%) and the most rapid and uniform germination were observed for all the lots when they were warm stratified for 56 days. Warm stratification improved germination more than alternate temperature stratification, while cold stratification inhibited germination especially for the 08Fr and ARg lots, thus seeds seem not to have a morphological component to their dormancy. GA₃ only improved germination of 07Fr seeds, at a low rate. A. acutifolius seeds fit the characteristics of a non-deep physiological dormancy.     

Temperatures affecting embryo development and seed germination of Christmas rose (Helleborus niger) after sowing

Embryo development followed by seed germination was investigated in seeds of Christmas rose (Helleborus niger) held under different temperature conditions. The seeds had a heart-shaped embryo when dispersed from parent plants. That embryo developed rapidly to the torpedo stage at 25 °C, after which no further development occurred. When seeds with a torpedo-stage embryo were held at 15 °C, the embryos developed to the cotyledon stage, but no further germination occurred. Similar phenomena in the course of embryo development were observed in seeds held in a non-heated polyethylene house, in which the embryos developed to the torpedo stage at 20 °C or more, while further development to the cotyledon stage was attained only at temperatures of 15 °C or less. Seeds with a cotyledon-stage embryo rarely germinated until they were treated at 4 °C for more than 8 weeks. These results suggest that seeds of H. niger have a deep, simple morphophysiological dormancy caused by the combination of rudimentary embryos and a physiological dormancy that can be broken by cycles of warming and chilling.     

Dormancy and germination in Rosa multibracteata Hemsl. & E. H. Wilson

Most of the achenes produced by Rosa multibracteata Hemsl. & E. H Wilson are dormant on maturity and require pretreatment to stimulate germination. To investigate the mechanism of dormancy and to develop effective methods of improving germination, roles of the pericarp, testa, and embryo of R. multibracteata in regulating dormancy were studied by investigating the effect of different pretreatments on germination. The effects of temperature and water stress were also tested with achenes treated by warm plus cold stratification. In freshly harvested achenes, pericarps are permeable and the embryo fully developed, which eliminates the possibility of physical, morphological, or morphophysiological dormancy. Germination percentage remained low (<5%) despite softening the pericarp or even removing it fully. However, fully removing the testa improved germination significantly (39%), indicating the possible presence of germination inhibitors in the testa. Dry storage, scarification with sulphuric acid (H₂SO₄), and warm stratification proved ineffective by themselves but when combined with cold stratification, improved germination and shortened the cold stratification period needed to break dormancy. Dry storage for 68 weeks followed by cold stratification for 16 or 24 weeks resulted in maximum germination (72–79%) among all the treatments. In achenes scarified with H₂SO₄, germination increased with an increase in the duration of cold stratification. Neither gibberellic acid (GA₃) nor ‘smoke water’ (water through which smoke had been bubbled for 2 h) had any positive effect on germination even on seeds that had been mechanically scarified or stratified. Both high temperature and water stress lowered germination in achenes treated with warm plus cold stratification. Our results suggest that R. multibracteata achenes have an intermediate physiological dormancy, and that dry storage for 68 weeks followed by cold stratification for 16 or 24 weeks is the best method for propagating R. multibracteata from seed.     

Methods to break seed dormancy in Cyclocarya paliurus (Batal)Iljinskaja

Cyclocarya paliurus is native to China and is the sole species in its genus. However, the seeds remain deeply dormant for 2 years in their natural environment. We tested different pretreatments of chemical scarification and exogenous application of gibberellic acid (GA₃) for efficacy in breaking dormancy and speeding germination. In contrast to scarified seeds, non-scarified seed did not germinate, indicating that C. paliurus seeds have hard, impermeable seed coat dormancy. Exogenous application of GA₃ significantly enhanced germination of scarified seeds. Compared with seeds stratified in sand with water, the germination of seeds stratified in sand moistened with 400 ppm GA₃ for 60 days was significantly increased and germination rate was over 90% after 120 days. Analysis of variance indicated that both GA₃ concentration and stratification medium had significant effects on seed germination and final germination percentage. Germination was higher for longer stratification periods, but no significant difference in germination was observed after 90 days. These results suggested that C. paliurus seeds exhibit both exogenous and endogenous dormancy. A combination of chemical scarification and exogenous application of GA₃ alleviated seed dormancy in a relatively short period of time.     

Effects of seed maturity,seed storage and pre-germination treatments on seed germination of cleome (Cleome gynandra L.)

The influence of seed maturity, seed storage and germination pre-treatments on seed germination of cleome (Cleome gynandra L.) were investigated. Seed maturation studies showed that capsules harvested at 18 days after anthesis possessed the highest dry weight with 19.2% moisture and 1% germination. Development of fresh-ungerminated seed was observed with increasing maturity of fruit, suggesting that cleome exhibited forms of seed dormancy. Storing mature seed at 15 °C and at room temperature for 5 months showed that seed dormancy was broken after 3 months under both storage regimes. When mature seeds were subjected to different treatments including various levels of GA₃, KNO₃, leaching, pre-chilling, soaking and pre-heating at different temperatures, it was found that pre-heating at 40 °C for period of 1–5 days was the most effective method in breaking dormancy in cleome.     

Promotion by 5-aminolevulenic acid of pepper seed germination and seedling emergence under low-temperature stress

The effects of incorporating 5-aminolevulenic acid (ALA) into the priming solution on low-temperature germination and emergence percentage performance of red pepper (Capsicum annuum cv. Sena) seeds before and after seed storage were investigated. Seeds were primed in 3% KNO₃ solution for 6 days at 25 °C in darkness containing 0 ppm, 1 ppm, 10 ppm, 25 ppm, 50 ppm or 100 ppm ALA. Following priming, seeds were either immediately subjected to germination and emergence tests at 15 °C or stored at 4 °C or 25 °C for 1 month after which they were subjected to germination and emergence tests at 15 °C. Priming pepper seeds in the presence of ALA improved final germination percentage (FGP) and germination rate (MGT) at 15 °C compared to non-primed seeds. The highest FGP was obtained from seeds primed in the presence of 25 ppm and higher ALA concentrations while the highest MGT was obtained from seeds primed in KNO₃ supplemented with 10 ppm ALA. Emergence percentages were the highest for the seeds primed in the presence of 25 ppm ALA and 50 ppm ALA while non-primed seeds had the lowest emergence percentage. Highest emergence rates (MET) and heaviest seedlings were also obtained from seeds primed in KNO₃ supplemented with 50 ppm ALA. Although all priming treatments improved germination and emergence performance of pepper seeds at 15 °C following 1 month of storage under two different temperatures, inclusion of 25 ppm and 50 ppm ALA into the priming solution resulted in higher germination and emergence percentages and faster germination and emergence compared to seeds primed in KNO₃ only and non-primed seeds. These results indicate that priming seeds in 25 ppm and 50 ppm ALA incorporated into the KNO₃ solution can be used as an effective method to improve low-temperature performance of red pepper seeds and that these seeds can be stored for 1 month at 4 °C or 25 °C and still exhibit improved germination and emergence performance at 15 °C.     

不同处理方法对莼菜种子发芽的影响

以当年采收的莼菜种子为材料，运用低温、激素、热水浸泡、不同发芽温度以及光照时间等
对莼菜种子进行处理，分析不同处理方法对其发芽率和发芽势的影响。结果表明：4 ℃低温处理对提高莼
菜种子发芽率有较好的效果，莼菜种子4 ℃水藏后用500 mg·L^-1 GA₃ 溶液浸泡6 h+40 ℃水浸泡50 min
对打破莼菜种子休眠有较好的作用；发芽温度以25 ℃最好，20 ℃条件下种子发芽周期较长，30 ℃条件下
种子发芽孔处易滋生白毛腐烂；种子在部分光照条件下发芽率较高，全黑暗条件下几乎不发芽。     

Effects of development,temperature, and calcium hypochlorite treatment on in vitro germinability of Phalaenopsis seeds

There are no standardized procedures for sanitizing orchid seeds for propagation by tissue culture and there is insufficient information about the optimum stage of orchid seed development for best germination. Phalaenopsis amabilis flowers were hand-pollinated and fruits harvested 90, 105, and 120 d after pollination (DAP) for seed developmental analysis. Embryo cell number per seed was counted after staining with 4′-6-diamidino-2-phenylindole and viewing through a confocal microscope. Germination percentage and cell number per embryo increased from 14 to 61% and 41 to 66%, respectively, during fruit development from 90 to 120 DAP. Seeds from mature, browning (∼140 DAP) Phalaenopsis Sogo Lit-Angel and Phalaenopsis spp. breeding line 9450 seed pods failed to germinate until frozen at −196, −80, or −18 °C and thawed or chilled at 4 °C for 10 d. Germinability in 140 DAP seeds was correlated with cracked testa after freezing and thawing. P. amabilis seeds were treated with 0, 5, 10, or 15% calcium hypochlorite (CH) for 5, 10, or 15 min. Ninety six percent of untreated seeds from 90 DAP fruit produced protocorms within 40 d after sowing (DAS). Exposing seeds to 5% CH for 10 or 15 min decreased germination to 85 and 73%, respectively. Exposure to 10 or 15% CH for 5, 10, or 15 min produced seed germination percentages of less than 40%. Protocorms developed root hairs and shoot primordia by 50 DAS and an average of one leaf and root by 85 DAS after treatment with either 0 or 5% CH. Higher concentrations delayed or inhibited protocorm development. Green fruits 120 DAP produced the highest percentage of protocorms, while ∼140 DAP seeds from browning fruit were dormant but cold treatments increased germination.     

Influence of harvest time and after-ripening on the seed quality of eggplant

Eggplant cv. Emi and Tsakoniki were cultivated for seed in an unheated greenhouse and fruits were harvested at 25–65 days after anthesis (DAA) in order to determine the optimum harvest time. In addition, the effect of after-ripening on seed quality (i.e. seed size and germination) was examined by storing harvested fruit at 25 °C for 20 days prior to seed extraction. From the results, it was concluded that the optimum time of harvest for seed production is 55 DAA. Seeds extracted from fruits that were harvested at 25–35 DAA did not germinate, but when fruits harvested at the same age were stored for 20 days at 25 °C prior to seed extraction (i.e. seeds were after-ripened) germination was induced. Seeds extracted from fruits harvested at 45 DAA showed a high percent germination, which decreased after storage at 25 °C for 3 months. This decrease, however, was reduced by after-ripening prior to extraction. It is concluded that although eggplant is a non-climacteric species and fruit do not ripen after harvest, nevertheless seeds within the fruit continue to fill and mature after harvest; hence storage of prematurely harvested fruit prior to seed extraction permits the seeds of these fruits to after-ripen in situ and thereby increases seed size and germination. The implication of this result for eggplant seed production is discussed.     

Physical dormancy in myrtle seed

The nature of dormancy in seeds of myrtle (Myrtus communis L.) was investigated. Scarification with cold acid or sand paper and soaking in water increased seed germination, whilst stratification, inserting the seeds in boiling water or hot acid treatment decreased the number of germinating seeds. The highest germination was obtained by treating the seeds with 100 % cold acid for 60 min or 80 % cold acid for 120 min. Hard seed coat was found to be the principal cause of poor seed germination.Scarified and non-scarified seeds were germinated for 2 weeks at 6 temperatures between 5 and 30° C, and 20° C was optimum.     

Dormancy and germination of Areca triandra seeds

The dormancy mechanisms of Areca triandra Roxb. Ex Buch-Ham seeds were studied by treating the intact or mechanically scarified seeds with scarification, chemical soaking and stratification. The results indicate that the seeds have exogenous and endogenous dormancy. The exogenous dormancy is imposed by the pericarp and it is the major limiting factor for germination. It can be broken by mechanical scarification, but not by chemical scarification in 98% H₂SO₄ for 30 min. Chemical treatments (soaking for 24 h in 100–200 mg/L GA₃, 0.2% KNO₃ and 0.1–0.3% NaNO₂, and for 12 h in 10% H₂O₂ or 20 min or 12 h in 15% H₂O₂) and stratifications, especially, cold stratifications for 30–120 days, broke the endogenous dormancy and significantly hastened germination of mechanically scarified seeds, although they did not increase the germination percentages.     

Seasonal periodicity of the loss of dormancy of imbibed seeds in Brassica vegetables

The present study deals with the mode of loss of dormancy of imbibed seeds in Brassica japonica and B. cernua. Mature seeds were sown in petri dishes immediately after harvest and were placed either at room temperature or at a constant temperature of 25°C. After the first flush of germination of non-dormant seeds, most seeds continued to be in a non-germinating state, suggesting that they were dormant. The germination of such dormant seeds occurred sporadically during several years. Under constant 25°C, the germination behaviour of the imbibed seeds was “quasi-simultaneous” in B. japonica and “continuous” in B. cernua. Both species showed a unimodal distribution of germination in time. Under fluctuating room temperature, the distribution of seed germination in each species was characterized by 3 peaks which occurred in summer. In B. japonica the 3 peaks were the same in height, but in B. cernua the middle peak was much higher than the other 2. Such seasonal periodicity of germination was caused by environmental factors of which temperature was the most important.     

Biocontrol of mildew with Bacillus subtilis in bitter gourd (Momordica charantia L.) seeds during germination

The bitter gourd seed has a thick, hard seed coat. Mildew often occurs during germination and causes uneven and low rates of seed germination. However, the problems caused by mildew can be overcome by treating seeds with hot water, by soaking in water, or by using microorganisms. Seeds of the ‘Ching Pi’ bitter gourd were treated in a water bath at 60 °C for 10 min and then soaked in tap water at 25 °C for 24 h. The resulting germination percentage was 86.7%, and the resulting percentage of mildewed seeds was 10%. The biocontrol potential of three commercially available Bacillus subtilis solutions was examined. For seeds primed with Huodijun B. subtilis solution, the germination percentage was 73.3% and the mildewed percentage 6%. In dual cultures, the antibiotic content in the Huodijun B. subtilis solution was significantly greater than in Yunghsing and Huolibao, the other B. subtilis solutions examined. B. subtilis effectively inhibited the growth of Rhizoctonia solani mycelium and caused abnormal mycelial growth.     

Effect of stratification temperature,stratification period and seed coat on seed germination of peach cultivar ‘Sharbati’

Freshly harvested seeds of peach cultivar ‘Sharbati’ were found to be dormant and did not germinate at all. A specific low-temperature stratification treatment was required to overcome seed dormancy. 10° C stratification was found to be the best for breaking seed dormancy. Increased seed germination was recorded when the seeds were after-ripened without seed coats as compared to the seeds after-ripened with seed coats. There was no further increase in germination when seeds without seed coats were stratified beyond 60 days at 10° C, while a significant increase was recorded up to 75 days of stratification in the case of seeds with seed coats. Unstratified seed coats, soaked in water for 72 h, leached out a water-soluble inhibitor, which could suppress the germination of stratified peach seeds without seed coats. When this leachate was bioassayed by a cress-seed germination test, it showed the presence of an inhibitor at Rf 0.7-0.9. Hence, delayed germination of seeds with seed coats may be due to the presence of an inhibitor in seed coats.     

Germination of Passiflora mollissima (Kunth) L.H.Bailey, Passiflora tricuspis Mast. and Passiflora nov sp. seeds

Passiflora mollissima, Passiflora tricuspis and Passiflora nov sp. are three passion fruit species occurring in Bolivia. Germination percentages and rates were determined for 11 different treatments. Per species, germination of 100 seeds was monitored every 3 days, during 90 days. Germination started after 9 days and 50% of final germination was reached within a month or less. Successful, recommended methods for P. mollissima are removing the basal point of seeds (27% germination) or removing the basal point in combination with pre-soaking seeds for 48 h in 50 ppm GA₃ (18%). Pre-soaking seeds for 24 h in 400 ppm GA₃ (42%) and removal of the basal point in combination with pre-soaking seeds for 48 h in 50 ppm GA₃ (36%) are suggested methods to improve germination of P. nov sp. Removing the apical point of P. tricuspis seeds resulted in maximal germination (57%). No unique treatment gave satisfactory results for the three species tested. Exogenous dormancy, probably a combination of mechanical and chemical dormancy is present in the three species studied. Presence of physical dormancy was found in P. mollissima.     

In vitro rescue of immature avocado (Persea americana Mill.) embryos

An in vitro culture protocol was developed as a means of avocado embryo rescue. Different factors including presence of cotyledons, medium texture and cold or gibberellic acid pretreatments, were studied. To better understand the germination process in this recalcitrant species, immature zygotic embryos at different stages were used in these experiments. Optimum results were dependant on the embryo developmental stage. Whereas smaller embryos (5 mm long) germinated better in M1 liquid medium, 15 mm long embryos responded better when precultured in B5m medium supplemented with 1 mg l⁻¹ GA₃, and fully mature embryos were capable of germinating directly in solid M1 medium. Our results suggest the existence of two types of dormancy in avocado embryos: an embryo-dormancy caused by cotyledons, and another type of dormancy, mainly occurring in 35 mm long embryos and revealed by the formation of dwarfing rosette seedlings, that can be released by a GA₃ pretreatment.     

月季杂交种子快速催芽方法初探简

通过研究不同时间低温层积、不同浓度萘乙酸（NAA）和不同浓度浓硫酸（H2SO4）对3个月季杂交种子发芽率的影响,探讨打破月季杂交种子休眠、快速催芽的有效方法。结果表明：（1）影响月季杂交种子萌发的最主要因素是低温层积时间,其最适层积时间为4℃沙藏80～100 d。（2）NAA和浓H2SO4浸种处理能有效打破休眠、提高发芽率,最佳浓度分别为50 mg/L和0.1%。