refine the standard seed germination protocol, in vitro seed germination methodology, and vegetative propagation techniques, including in vitro multiplication of cloned plantlets, to facilitate ex situ conservation and development of a new methodology for
Phillip A. Wadl, Timothy A. Rinehart, Adam J. Dattilo, Mark Pistrang, Lisa M. Vito, Ryan Milstead, and Robert N. Trigiano
Joseph J. King and Mark P. Bridgen
Presowing treatments and temperature regimes were tested to improve germination of Alstroemeria hybrids 3 to 12 months following harvest. In addition, seeds from 20 intraspecific F1 hybrids of five selections were also tested 3 to 7 or 8 to 12 weeks following harvest. Seeds were pretreated by chipping the seedcoat above the embryo, general abrasion of the entire seedcoat, or soaking 12 hours in distilled water, GA, (0.029, 0.29, 2.9 mm), or KNO3 (0.5 and 1.0 m). Pretreatments were evaluated under three environmental regimes: 8 weeks at a constant 18-25C (warm), 4 weeks at 18-25C followed by 4 weeks at 7C (warm-cold), or 4 weeks at 7C followed by 4 weeks at 18-25C (cold-warm). There was an interaction between pretreatment and environmental regime for percent germination. Germination percentages for the water soak and GA, at 0.29 or 2.9 mm were significantly higher than for the other pretreatments, but were not significantly different from one another. The warm-cold environment yielded higher germination percentages than the other environments. The time to germination was longest for the cold-warm regime. This response depended on the genotype and the age of the seed. Chemical name used: gibberellic acid (GA3).
Stanta Cotner, John R. Clark, and Eric T. Stafne
A study was conducted in the Winter–Spring 2004 to evaluate the effects of seed (pyrene) scarification period on blackberry (Rubus L. subgenus Rubus) genotypes that had a range of seed weights. The study was done in an attempt to identify optimum scarification period for variable seed weights for the purpose of increasing germination of blackberry seeds produced from hybridizations in the Arkansas blackberry breeding program. Scarification treatments of 1, 2, or 3 hours were used on 14 genotypes. Seeds were then stratified for 3.5 months and sowed on a commercial potting medium in a heated greenhouse. Germinating seedlings were counted over a 15-week period and total germination determined. Data analysis indicated significant genotype effect on germination but no scarification treatment nor genotype × scarification treatment interaction significance. The results indicated that scarification period did not affect germination and varying this period predicated on seed weight was not beneficial based on the genotypes used in the study.
Amy Douglas and Rosanna Freyre
Nolana is a diverse genus native to coastal deserts of Peru and Chile, with great potential for developing new ornamental cultivars. Low germination has been an obstacle to breeding efforts at the University of New Hampshire (UNH). Nolana fruits are comprised of unusual sclerified mericarps, each containing one or more embryos. Germination occurs with opening of funicular plugs on the mericarps. Under normal greenhouse conditions at UNH, germination success in eight Nolana species (N. adansonii, N. aticoana, N. humifusa, N. laxa, N. ivaniana, N. plicata, N. elegans, and N. rupicola) ranged from 0 to 0.05 seedlings/mericarp. We analyzed mericarp morphology, imbibition, and the effect of chemical and environmental germination treatments. SEM showed that soaking treatments create physical changes in mericarp morphology, exposing tracheid tubes in the funicular plugs. Mericarps were soaked in dye to track imbibition, confirming that this occurs through the tracheid tubes, and that additional scarification is not required. The following chemical treatments were unsuccessful in increasing germination: 0.1 N HNO3, 0.2 KNO3, conc. H2SO4, 10 mM or 1 μM ethephon. Gibberellic acid (1000 ppm) effectively increased germination in some species (up to 0.47 seedlings/mericarp). Mericarps stored dry for 2 years had significantly higher germination than fresh mericarps (0.55 seedlings/mericarp). Mericarps of N. aticoana were subjected to after-ripening treatments. Mericarps stored for 7 weeks at 35 °C and 75% RH showed significantly higher germination (0.36 seedlings/mericarp) than mericarps stored dry, or stored moist for 1-6 or 8-12 weeks. Our findings facilitate development of larger hybrid populations, thus increasing the efficiency of Nolana breeding programs.
Augusto Trejo-Gonzalez and Marita Cantwell
A lime solution is an efficient starch gelatinization agent during the traditional process of nixtamalization of corn for tortilla production. Although the saturated Ca(OH)2 solution is usually heated to speed the process, similar physical-chemical changes occur to corn steeped at room temperature. In addition, the steeped grains are capable of rapid germination. In this study we explored whether, in barley grains subjected to the same steeping conditions, there would be an acceleration of biochemical changes for the production of malt. Barley grains cv. Esmeralda six-row were steeped in saturated solutions of lime from 0% to 2% (w/w) at 18 to 23 °C for up to 72 hours. After treatment, the grains were washed with water and placed in a germination cabinet at 20 ± 2 °C and 90% RH for up to 96 hours to germinate. Activity of α-amylase (U/mg protein), sprout length (cm), seed viability (tetrazolium test), and respiration rates were determined. Scanning electron micrographs were prepared. There was a very rapid uptake of lime solution by the barley grains during the first 30 hours of treatment. The barley grains were 98% viable after all periods of steeping. Grains steeped 24 hours and germinated 70 hours had the highest activity of α-amylase, longest sprout length and highest percentage of germination. Under SEM, chemical gelatinization of starch grains was observed in barley sections after different steeping treatments, and starch breakdown was observed in grains during germination. No fungal growth was observed during germination after the steeping treatments. These results demonstrate that steeping in lime solutions could shorten the period required for barley germination and similar conditions may be useful for germination of other cereal and vegetable seeds.
Frank A. Blazich, Paul H. Henry, and Farrell C. Wise
Seeds of `Dawn Carpet' and `Little Bright Eye' annual vinca [Catharanthus roseus (L.) G. Don] were subjected to 32 treatments, arranged as a four × four × two factorial. For each cultivar, seeds were exposed to one of four temperatures (15, 20, 25, or 30C) during the 8-hour (day) and 16-hour (night) portions of the cycle. Within each temperature regime, half the seeds of each cultivar were irradiated for 1 hour daily, and the other half remained in constant darkness. Final germination percentages were suppressed at 15C day or night temperatures; at temperatures ≥20C, there were no significant differences between treatments. Heat input (daily degree hours) was a controlling factor in germination; different temperature cycles with equivalent numbers of daily degree hours had similar effects on germination response. There was a strong interaction between temperature and irradiation regime for both cultivars. Irradiating seeds for 1 hour/day reduced final germination percentages under cool (15C) conditions; response was not adversely affected when seeds at 15C were germinated in darkness. In a second experiment, seeds at 25C were exposed to daily photoperiods of 0, 1, 2, 4, 8, 12, or 24 hours. Germination percentages obtained in darkness and at photoperiods ≤12 hours were equivalent. Twenty-four-hour photoperiods suppressed germination compared to all other irradiation treatments.
Tim D. Davis, Daksha Sankhla, N. Sankhla, A. Upadhyaya, J.M. Parsons, and S.W. George
Seeds of Aquilegia chrysantha Gray were germinated under a variety of temperature regimes. Germination was nearly 90% under a day/night cycle of 25/20C, but was reduced to ≤ 40% under constant 25C or a 25/10C day/night cycle. With days between 25 and 29C (night = 20C), germination percentage dropped gradually to ≈ 60% with increasing temperature. With days >29C, germination declined dramatically such that no germination occurred at 31C. Neither kinetin (4.6 to 46 μm) nor ethephon (6.9 to 207 μm) was able to reverse the inhibitory effects of 33C days. Our results indicate that germination of A. chrysantha seed is sensitive to temperature and that germination ≈ 75% can be obtained under a 25 to 27C day/20C night regime. Chemical names used: 2-chloroethylphosphonic acid (ethephon); 6-furfurylaminopurine (kinetin).
Samuel Contreras, David Tay, and Mark Bennett
Lettuce seeds (Lactuca sativavar. acephalacv. Tango) were used with the objective of determining the effect of temperature, light, and their interactions in promoting germination. Under standard op-timal conditions (20 °C, light), the seed presented 100% germination (radicle emergence 5 d after sowing). Different treatments evaluated germination under dark conditions, with or without a red light break (LB, 28.8 mmol·m-2) 48 h after sowing, and with different combination of temperatures pre- (soaking temperature, ST) and post- (germination temperature, GT) the LB. Germination at constant 20 °C without LB was less than 5%, and with LB, it was around 30%. However, germination was close to 100% at GT of 20 °C when LB was applied after a ST of 10 °C, and around 50% under the same conditions, but without LB. When GT was 30 °C and LB was applied, germination was less than 3% with ST = 30 °C, less than 10% with ST = 20 °C, and around 100% when ST = 10 °C. With ST and GT of 10 °C and 30 °C, respectively, and no LB, germination was less than 5%. Germination at 10 °C constant, with and without LB, was around 90% and 0%, respectively. When ST was 40 °C and LB was applied, germination was around 40% at GT= 20 °C, but less that 3% with GT= 30 °C. In summary, a severe inhibition of germination was observed when seeds were germinated in dark, which was partially reversed by either a light treatment or soaking at 10 °C, and fully reversed when both treatments were applied together. Inhibition of lettuce germination at 30 °C was observed when this temperature was applied after a light treatment, but not when applied before. Possible implications of these results for the phytochrome mechanism of action are discussed.
Ikuo Miyajima, Adriana Kato, Juan Carlos Hagiwara, Diego Mata, Gabriela Facciuto, Silvina Soto, Alejandro Escandón, Marcela Mori, and Nobuo Kobayashi
In vitro germination of immature seeds of Jacaranda mimosifolia treated with gibberellic acid (GA3) was studied. Immature seeds were collected monthly after crossings and sown on Murashige and Skoog (1962) medium with 3.0% sucrose and 0.6% agar after soaked 24 hours with 0, 10, 100, and 500 mg·L–1 GA3 solutions. Though germination was observed in the immature seeds harvested 2 months after crossing (2 MAC), the rate was quite low. When immature seeds of 3 MAC treated with 100 or 500 mg·L–1 GA3 solution were cultured, >60% germination were obtained within 2 weeks after culturing. These results indicate that immature seeds of 3 MAC treated with adequate GA3 solutions, seedlings can be obtained precociously and the period from crossing to the seedling stage was shorter than for mature seeds.
Phil S. Allen and Susan E. Meyer
To determine optimum germination temperatures and effective dormancy-breaking procedures, field-grown (1983-85) seeds of `Bandera' Rocky Mountain penstemon (Penstemon strictus Benth), `Cedar' Palmer penstemon (Penstemon palmeri Gray), and firecracker penstemon (Penstemon eatonii Gray) were subjected to various cold stratification and incubation temperature treatments. Increased germination following an 8-week stratification occurred in seed lots containing dormant seeds, but a 2-week stratification generally failed to break dormancy. Older (1983) seeds of `Bandera' and `Cedar' penstemon germinated to full viability without stratification. All species showed a marked decrease in germination percentage above 20C; 15C consistently produced maximum germination after 4 weeks. At 15C, mean times to 90% of total germination were 11, 22, and 29 days for `Bandera', `Cedar', and firecracker penstemon, respectively. Transfer of seeds failing to germinate at warm temperatures (25 and 30C) to 15C and applying 720 μm gibberellic acid (GA3) solution was effective in breaking primary dormancy of firecracker penstemon and secondary dormancy of `Bandera' penstemon. Our findings suggest that incubation below 20C, combined with 8 weeks of stratification or the use of after-ripened seed, may improve seed propagation efforts for these species.