Few studies on embryogenesis in common bean (Phaseolus vulgaris L.) have been reported and only the early stages of somatic embryogenesis were observed. Dry seeds from two common bean lines were germinated in darkness on L-6 medium containing 4% sucrose, 0.2 g casein hydrolysate /liter and 2.0 g phytagel /liter. The medium for seed germination was supplemented with 0, 2, 4 or 6μM forchlorfenuron (CPPU). Explants from cotyledonary leaves, petioles, hypocotyls and shoot apices were prepared from 14 day-old seedlings. Callus was derived from explant cultures incubated in darkness at 26C on the medium containing 4 μM 2,4-D and 1 μM Kinetin. The callus was transferred after 4 weeks into 125 ml Erlenmeyer flasks containing 50 ml liquid medium and placed on a gyrotary shaker (120 rpm) under cool-white light (12 μmol.m-2 .s-1 ). The liquid medium was used with 2, 4 or 6 μM of 2,4-D alone or with zeatin supplements at relative concentrations of 0.25 and 0.5. Up to 200 somatic embryos from 40 to 50 mg callus inoculations were induced after 4 to 5 weeks. Callus derived from seedlings grown on CPPU-containing medium gave more repetitive somatic embryos. Cotyledonary stage embryos with clear bipolar structure were observed only from callus derived from seedlings grown on CPPU when transferred to suspension cultures containing 2,4-D and zeatin. All somatic embryos differentiated strong roots and some developed leaf-like structures on conversion medium.
Seeds of celery, spinach, onion, cress, water cress, iceberg lettuce, Great Lakes lettuce, cabbage, tomato, sweet corn and celery were pre-treated with 0.1 μM/g seed of both ABA and analogs of ABA. The chemicals were dissolved in a mixture of methanol:hexane (9:1/v:v) and applied to the seeds for approximately 3 minutes. The solvent was removed from the seeds within 5 minutes by rotary evaporation under reduced pressure. Effects on petri plate germination and soil emergence were monitored daily at 5, 10 and 15°C. The methanol/hexane solvent alone improved spinach seed emergence at 10°C from 10% to 100% and from 50% to 90% at 15°C in celery. Certain ABA analogs reduced time to 50% emergence in celery by at least 7 days at 15°C. Two ABA analogs synchronized emergence in celery and effect was temperature-dependent. One analog improved seed germination in tomato from 15% to 90% at 10°C. In most cases treatment effects on radicle germination on petri plates was not a good indicator of treatment effects on emergence from a soil based system.
Tillandsia eizii is an epiphytic bromeliad that due to over-collection, habitat destruction, and physiological constraints has declined to near threatened status. This species exhibits high mortality in the wild, and seed are characterized by low percentages of germination. As a means to conserve this species, in vitro culture protocols were developed to enhance seed germination and seedling growth. A sterilization protocol using 70% ethanol for 2 minutes followed by 2.6% NaOCl for 40 minutes disinfested seed and promoted seedling growth. Sucrose incorporated into the culture medium had no effect on germination or growth, while NAA inhibited growth, but not germination. Cultures maintained under a 16-hour photoperiod at 22 °C exhibited greater growth than those grown at 30 °C. Seed that germinated in the dark remained etiolated and failed to develop even after transfer to light conditions. Plants grown in vitro were successfully acclimatized and transferred to the greenhouse. Over 86% survival and rapid growth were obtained with either an all-pine-bark medium, or a mixture of 2 redwood bark: 2 fir bark: 2 potting mix: 1 perlite. This demonstrated that in vitro culture of seed may be used to rapidly produce large numbers of T. eizii, and thus can be used for the conservation and reintroduction of this species.
Seeds of six provenances (Escambia Co., Ala.; Santa Rosa Co., Fla.; Wayne Co., N.C.; Burlington Co., N.J.; New London Co., Conn.; and Barnstable Co., Mass.) of Atlantic white-cedar [Chamaecyparis thyoides (L.) B.S.P.] were stratified (moist-prechilled) for 0, 30, 60, or 90 days at 4 °C. Following stratification, seeds were germinated at 25 °C or an 8/16-hour thermoperiod of 30/20 °C with daily photoperiods at each temperature of 0 (total darkness), 1, or 24 hours. The germination of nonstratified seed did not exceed 18%. Seeds germinated at 25 °C required 60 to 90 days stratification to maximize germination. In contrast, 30 days stratification maximized germination at 30/20 °C. Regardless of stratification duration, germination was generally lower at 25 °C than at 30/20 °C for each provenance. Averaged over all treatments, seeds of the Alabama provenance exhibited the greatest germination (61%), followed by those from Florida (45%), with the remaining provenances ranging from 20% to 38%. However, specific treatments for each provenance induced germination >50%. Germination of seeds not exposed to light was <8%, in contrast with 48% and 55% germination for daily photoperiods of 1 and 24 hours, respectively. Seeds from each of the provenances, except for Alabama, exhibited an obligate light requirement when germinated at 25 °C. At 30/20 °C, the North Carolina, New Jersey, Connecticut, and Massachusetts provenances required light for germination, whereas the Alabama and Florida provenances did not.
Little scientific information is available describing morphological development of pawpaw during seed germination. To provide this information, a study was designed to outline important developmental stages and describe seedling characteristics within each stage. Stratified pawpaw seeds were sown in vermiculite and germinated at 25°C in a growth chamber. Ten seedlings were randomly chosen and destructively harvested at 5-day intervals starting at radicle protrusion. Length (mm), fresh and dry weight, and percentage of total dry weight were determined for seedling components. Pawpaw seeds have a small rudimentary embryo with all food reserves stored in a ruminate endosperm. Dry weight measurements showed a dramatic reallocation of reserves from the storage tissue to developing seedling parts. Initial embryo length was less than 3 mm, but within 70 days seedlings exceeded 350 mm. Twelve days after planting, simultaneous radicle and cotyledon growth occurred (3.4 and 3.0 mm, respectively), but neither hypocotyl nor epicotyl was visible. Radicle protrusion was observed at 15 days with radicle, cotyledon and hypocotyl lengths increasing to 4.4, 4.0, and 3.2 mm, respectively. Endosperm comprised 99.1% of total dry weight at this stage. The hypocotyl hook emerged after 30 days and endosperm comprised 76.1% of total dry weight. Cotyledons reached maximum length (29.0 mm) at day 40 and the epicotyl was discernible. At 55 days, the seed coat containing cotyledons and residual endosperm abscised and the average radicle, hypocotyl and epicotyl lengths were 182.0, 61.1, and 7.3 mm, respectively. It is suggested that the cotyledons primary function is absorption of food reserves from the endosperm for transfer to the developing seedling.
Genotypic variation and horticultural potential of Alnus maritima [Marsh.] Nutt. (seaside alder), a large shrub or small tree found naturally in only three small, disjunct populations, have not been studied. We examined effects of population of origin and environment on seed germination and growth and morphology of seedlings. The first experiment showed that 6 weeks of cold stratification optimized germination of half-sibling seeds from Oklahoma at 73.2%. When this treatment was applied to multiple groups of half-siblings from all populations in a second experiment, seeds from Oklahoma had a higher germination percentage (55.0%) than seeds from Georgia (31.4%) and the Delmarva Peninsula (14.7%). In a third experiment, morphology and growth of multiple groups of half-siblings from all three populations were compared in one environment. Leaves of seedlings from Oklahoma were longer (12.8 cm) and more narrow (2.15 length to width ratio) than leaves of seedlings from Georgia (12.0 cm long; ratio = 1.76) and the Delmarva Peninsula (11.6 cm long; ratio = 1.86). Seedlings from Oklahoma and Georgia accumulated dry weight at higher rates (181 and 160 mg·d-1, respectively) than seedlings from Delmarva (130 mg·d-1), while seedlings from Oklahoma and Delmarva were more densely foliated (0.72 and 0.64 leaves and lateral shoots per centimeter of primary stem, respectively) than those from Georgia (0.46 per cm). These differences indicate genetic divergence among the three disjunct populations and the potential to exploit genetic variation to select horticulturally superior A. maritima for use in managed landscapes.
Temperature is an important environmental factor that affects lettuce (Lactuca sativa L.) germination. The present research was conducted to determine the role of seed coverings on lettuce seed germination at high temperature. Five lettuce genotypes were primed in order to bypass thermoinhibitional effects on germination. During germination of primed and nonprimed seeds, imbibition followed a normal triphasic pattern. Primed seeds had higher final water content, a decreased imbibitional phase II, and germinated at 36 °C compared to nonprimed seeds of thermosensitive genotypes, which did not germinate at 36 °C. Puncture tests were conducted to determine the force required to penetrate the whole seed or endosperm of the five genotypes at 24 and 33 °C. `Dark Green Boston', a thermosensitive genotype, had the highest mean resistance (0.207 N) and PI 251245, a thermotolerant genotype, had the lowest (0.139 N). Resistance to penetration of the endopserm of the five genotypes was different at both temperatures. However, three thermotolerant genotypes had lower endosperm resistance than two thermosensitive types. At 36 °C, the penetration force for primed and nonprimed seeds was compared after the first hour of imbibition and 1 hour before radicle protrusion. The force required to penetrate the seed was affected by genotype, seed priming, and duration of imbibition. Puncture force decreased as imbibition time at 36 °C increased in primed and nonprimed seed of each thermotolerant genotype but not in the thermosensitive genotypes. Priming reduced the initial force necessary to penetrate the seed and endosperm in all genotypes. Thus, for radicle protrusion to occur, there must first be a decrease in the resistance of the endosperm layer as evidenced by priming or thermotolerant genotype. Then, the pericarp and integument are sufficiently weakened so that tissue resistance is lower than the turgor pressure of the expanding embryo, allowing germination to be completed.
The effect of preplant conditioning on germination of three flower seeds, Bupleurum griffithii (Tourn.) L. (thorough-wax), Ammi majus L. (greater ammi), and Cirsium japonicum DC. Per. (Japanese thistle), were studied. Seeds were osmoconditioned with -1.2 MPa polyethylene glycol 8000 (PEG) solution and matriconditioned with moist Micro-Cel E (ratio of 2 seed: 0.6 carrier: 3 water by weight for Bupleurum and Cirsium; for Ammi the ratio was 2:1.4:6) and moist expanded vermiculite #5 (the ratio was of 2 seed: 0.6 carrier: 2 water for Bupleurum). In some treatments, water in the matriconditioning mixture was replaced with 1 mm gibberellin A4+7 (GA) or 0.2 % KNO3. In Bupleurum, matriconditioning with Micro-Cel E was generally superior to matriconditioning with vermiculite or osmoconditioning with PEG. A 4-day matriconditioning with Micro-Cel E and germination in the dark reduced the period required for 50% (T50 of final germination by 4 days and improved the percentage germination at 20C (73 % vs. 95%), compared to nonconditioned seeds germinated in the dark. The treatment also improved the percentage of germination at 15C (68% vs. 95%) and effectively removed the thermoinhibition of germination at 25 and 30C. Germination was inhibited to a greater extent for seeds kept in the light during matriconditioning and germination than for seeds conditioned in darkness and germinated in light or conditioned in light and germinated in darkness. Nitrate added during conditioning in light prevented inhibition of germination, provided seeds were kept in darkness during germination. In A. majus, germination in light after 4-day matriconditioning reduced the T50 by ≈2 days, but had little effect on percentage germination. Both GA and irradiance equally promoted germination when added during osmoconditioning, with nitrate having no effect. In C. japonicum, a 4-day matriconditioning or a 7-day osmoconditioning reduced the T50 of germination by -2 days and improved the percentage germination to some extent. Neither irradiance nor nitrate had any significant effect.
( Bernstein and Ayers, 1953 ; Maas and Hoffman, 1977 ). To date, there have been few evaluations of the salinity tolerance of carrot during the seed germination stage ( Kahouli et al., 2014 ; Rode et al., 2012 ; Schmidhalter and Oertli, 1991 ). These
( Zheng et al., 2013 ) and sticky snakeroot [ Eupatorium adenophorum (unpublished data)] could inhibit seed germination of mumian. However, seedling regeneration is still impaired around the mature trees even though no sticky snakeroot exists at some