Enhancement of Festuca rubra L. germination and seedling growth by seed treatment with pathogenic Agrobacterium rhizogenes

The study concerned effects of two methods of red fescue seeds treating with wild type Agrobacterium rhizogenes strains (15834 and LBA 1334): soaking and matriconditioning with Micro-Cel E solid carrier. The effects were assessed from germination and seedlings growth rates. The bacterial indole-3-acetic acid (IAA) production and ACC deaminase activity were also determined. Both strains are able to produce the IAA and showed ACC deaminase activity in various amounts. The strains accelerated seeds germination, seedling emergence and development. The beneficial effect on those processes was visible when seeds were soaked for 1 h in bacterial suspension and, especially, when the bacteria were present during Micro-Cel E conditioning. The main effect observed upon inoculation of seeds during priming was increased growth of lateral roots and more complex architecture of the branching root system. Treating seeds with A. rhizogenes simultaneously with Micro-Cel E priming as a carrier is a promising method of enhancing grass germination and seedlings growth, particularly by improving the root system development.     

Gamma radiation effects on seed germination, growth and pigment content, and ESR study of induced free radicals in maize (Zea mays)

The effects of gamma radiation are investigated by studying plant germination, growth and development, and biochemical characteristics of maize. Maize dry seeds are exposed to a gamma source at doses ranging from 0.1 to 1 kGy. Our results show that the germination potential, expressed through the final germination percentage and the germination index, as well as the physiological parameters of maize seedlings (root and shoot lengths) decreased by increasing the irradiation dose. Moreover, plants derived from seeds exposed at higher doses (≤0.5 kGy) did not survive more than 10 days. Biochemical differences based on photosynthetic pigment (chlorophyll a, chlorophyll b, carotenoids) content revealed an inversely proportional relationship to doses of exposure. Furthermore, the concentration of chlorophyll a was higher than chlorophyll b in both irradiated and non-irradiated seedlings. Electron spin resonance spectroscopy used to evaluate the amount of free radicals induced by gamma ray treatment demonstrates that the relative concentration of radiation-induced free radicals depends linearly on the absorbed doses.     

Effects of sand burial depth on the root system of Salix cheilophila seedlings in Mu Us Sandy Land, Inner Mongolia, China

Salix cheilophila Schneid. is a naturally occurring Salix species in Mu Us Sandy Land, Inner Mongolia, China. We focused on the morphological adaptability of S. cheilophila to sand dune burial. For morphological measurements, 32 S. cheilophila seedlings were removed from a community which was in the process of being buried by a shifting sand dune. Each seedling collected included the entire root system. We measured the number, length, and biomass of the adventitious roots, primary lateral roots, and taproot, and compared the morphological characteristics of the root system, including adventitious roots, for seedlings buried to various levels in the sand. The growth range of adventitious roots increased as the length of the buried portion of the main shoot increased. In addition, the total dry weight of all current-year shoots tended to increase gradually with increasing total dry weight of the adventitious roots. These results suggest that S. cheilophila tends to make use of the sedimentary sand layer that accompanies shifting sand dunes. However, there was no correlation between biomass or number of adventitious roots and the length of the buried part of the main shoot. Thus, S. cheilophila does not grow adventitious roots proportional to the buried part. These morphological characteristics of the root system, including the adventitious roots, may indicate that S. cheilophila has poor morphological adaptability to sand dune burial.     

Asymbiotic seed germination of Cymbidium bicolor Lindl. (Orchidaceae) and the influence of mycorrhizal fungus on seedling development

An in vitro plant regeneration protocol was successfully established for Cymbidium bicolor an epiphytic orchid by culturing seeds from green pods. Immature seeds were germinated on four basal media viz., Murashige and Skoog (MS) medium, Knudson C (KC) orchid medium, Knudson C modified Morel (KCM) medium and Lindemann orchid (LO) medium. Seed germination and protocorm development was significantly higher in LO medium (96.6 %) followed by KCM (89 %), MS (77.5 %) and KC (62.7 %) media after 56 days. For multiple shoot induction the protocorms were transferred to B5 medium supplemented with cytokinin. Among the various cytokinins tested, BAP (4.42 μM) induced maximum (27.59) number of multiple shoots per explant. IBA was effective in inducing healthy roots. Tissue-cultured protocorms and seedlings of C. bicolor were inoculated with AC-01 fungal strain (Moniliopsis sp.) isolated from the mycorrizal roots of an epiphytic orchid Aerides crispum. Mycorrhizal fungi significantly enhanced number of roots, root length and shoot number.     

Cryopreservation,early seedling development,and genetic stability of <Emphasis Type="Italic">Oncidium flexuosum</Emphasis> Sims

An efficient cryopreservation protocol was developed for mature seeds of Oncidium flexuosum Sims. Seed morphology, protocorm formation, and early seedling development were also assessed. The effects of phloroglucinol and Supercool X-1000^® as cryoprotectant additives in the vitrification solution were investigated. Dehydration using the plant vitrification solution 2 (PVS2) for 60 and 120 min prior to immersion in liquid nitrogen promoted the highest frequency of in vitro seed germination 6 weeks following culture on half-strength Murashige and Skoog (½ MS) medium. Mature seeds submitted to vitrification for 120 min in PVS2 and 1 % phloroglucinol at 0 °C enhanced germination by 68 %, whereas in PVS2 and 1 % Supercool X-1000^® germination was just moderately enhanced (26 %). In vitro-germinating seedlings developed healthy shoots and roots without the use of plant growth regulators. After 6 months of growth, there were no differences between in vitro- and ex vitro-grown seedlings for various phenotypic characteristics, including shoot length, number of leaves, number and length of roots, and fresh and dry weight. Seedlings were transferred to greenhouse conditions and successfully acclimatized, further developing into normal plants with over 90 % survival. Comparative analysis of seedlings from control and vitrified seeds using flow cytometry indicated that no change in ploidy levels occurred as a result of cryopreservation, therefore maintaining seedlings genetic stability. In this study, vitrification with PVS2 for 120 min with the addition of 1 % phloroglucinol offers a simple, safe, and feasible protocol for cryopreservation of O. flexuosum mature seeds.     

Cytokinin-producing, plant growth-promoting rhizobacteria that confer resistance to drought stress in Platycladus orientalis container seedlings

One of the proposed mechanisms through which plant growth-promoting rhizobacteria (PGPR) enhance plant growth is the production of plant growth regulators, especially cytokinin. However, little information is available regarding cytokinin-producing PGPR inoculation on growth and water stress consistence of forest container seedlings under drought condition. This study determined the effects of Bacillus subtilis on hormone concentration, drought resistance, and plant growth under water-stressed conditions. Although no significant difference was observed under well-watered conditions, leaves of inoculated Platycladus orientalis (oriental thuja) seedlings under drought stress had higher relative water content and leaf water potential compared with those of noninoculated ones. Regardless of water supply levels, the root exudates, namely sugars, amino acids and organic acids, significantly increased because of B. subtilis inoculation. Water stress reduced shoot cytokinins by 39.14 %. However, inoculation decreased this deficit to only 10.22 %. The elevated levels of cytokinins in P. orientalis shoot were associated with higher concentration of abscisic acid (ABA). Stomatal conductance was significantly increased by B. subtilis inoculation in well-watered seedlings. However, the promoting effect of cytokinins on stomatal conductance was hampered, possibly by the combined action of elevated cytokinins and ABA. B. subtilis inoculation increased the shoot dry weight of well-watered and drought seedlings by 34.85 and 19.23 %, as well as the root by 15.445 and 13.99 %, respectively. Consequently, the root/shoot ratio significantly decreased, indicative of the greater benefits of PGPR on shoot growth than root. Thus, inoculation of cytokinin-producing PGPR in container seedlings can alleviate the drought stress and interfere with the suppression of shoot growth, showing a real potential to perform as a drought stress inhibitor in arid environments.     

Evaluating the deterioration of peanut seeds contaminated with aflatoxin

Although aflatoxin (AFT) effects on Arachis hypogaea seed germination have been examined, there have been few, if any, investigations correlating germination capacity to field AFT levels for naturally contaminated peanuts. Here, we report this analysis for selected Federal samples containing 0–20 ppb AFT collected at intervals during both September and October 1981. Following the removal of both decoated and split seeds for screening, seeds were germinated for 5 days when both dead and severely infested seeds were discarded. The remaining seeds, normal (uninfested) ‘sprouted’, ‘sprouted’ with some infestation, ‘sprouted’ with morphological abnormalities and normal unsprouted, were both measured and transferred to fresh towels for germination and growth of seedlings. The hypocotyl and primary root lengths of the seedlings were measured. Whereas these lengths ranged from 58·0 ± 6·2 to 68·8 ± 5·6 mm for contaminated seedlings (control 72·5 ± 5·9) at 5 days, hypocotyl and root lengths were 210·0 ± 12·5–225·0 ± 23·0 for contaminated seedlings (control 226·7 ± 21·3) by day 12. Then, percentage germination was 75·0 ± 3·0−84·4 ± 2·5 for toxin seeds control 85·6 ± 1·8). These results suggest that neither the percentage germination nor hypocotyl/ root lengths could be strongly correlated with AFT levels (from natural contamination) for the selected seeds.     

Surface colonization of lodgepole pine (Pinus contorta var. latifolia [Dougl. Engelm.]) roots by Pseudomonas fluorescens and Paenibacillus polymyxa under gnotobiotic conditions

Indirect immunofluorescence techniques and confocal scanning laser microscopy were used to identify rhizobacterial strains on the root surfaces of pine seedlings, which were grown from seeds under gnotobiotic conditions. Conifer plant growth promoting rhizobacterial strains Paenibacillus polymyxa L6 and Pw-2, and the forest soil isolate Pseudomonas fluorescens M20, were inoculated onto surface-disinfested pine seeds, singly, or in dual combinations: strains L6 + M20, or strains Pw-2 + M20. Segments containing particular root microsites (root tip, root hair zone, or areas of lateral root emergence) were sampled randomly from roots 7 or 13 weeks after inoculation, and the colonization of roots by each bacterium was observed. Root segments were also sampled from individual roots at six different points along the length of the root, and the qualitative colonization of younger areas, closer to the root tip, contrasted with that of older areas, closer to the root base. The ability of strain M20 to colonize root areas adjacent to sites of lateral root emergence improves in the presence of either P. polymyxa strain, while the ability of the P. polymyxa strains to colonize these areas was not affected. More rhizobacteria were also generally observed on younger root tissues than on areas closer to the root base.     

Spore associated bacteria of arbuscular mycorrhizal fungi improve maize tolerance to salinity by reducing ethylene stress level

The role of spore associated bacteria of arbuscular mycorrhizal fungi (AMF) in improving plant growth and alleviating salt stress is a potential area to explore. In the present study, 22 bacteria isolated from the spore walls of AMF were identified to contain 1-aminocyclopropane-1-carboxylate deaminase. These were tested for their ability to improve seed germination and alleviate salt stress in the early growth of maize. Among the isolates, 19 bacteria that were able to grow at 4?% NaCl were used for germination assay. Two bacteria and seven bacteria significantly improved maize seed germination at 100 mM NaCl and 200 mM NaCl, respectively. Based on the presence of plant growth promoting (PGP) characters and the ability to improve seed germination, five strains were chosen for further experiments. At 0 mM NaCl, all the strains were able to increase maize shoot and root growth significantly. At 25 mM NaCl, except for Bacillus aryabhattai S210B15, all the strains were able to increase shoot and root growth significantly. At 50 mM NaCl, Bacillus aryabhattai S110B3 and B. aryabhattai S210B15 significantly improved shoot length, whereas, Pseudomonas koreensis S2CB35 and B. aryabhattai S210B15 significantly increased root length. Although salinity increased ethylene production in maize, bacterial inoculation significantly reduced the ethylene level at 0, 25 and 50 mM NaCl. Among the five strains, only P. koreensis S2CB35 showed the presence of PGP functional traits of nifH, acdS and nodA genes.     

Germination and Early Growth of Plants Studied Using Neutron Radiography

Seed swelling, germination, root extension, lateral root initiationand shoot growth were studied in soils of different water contents,using non-destructive, serial neutron radiography. Seeds fromthree varieties of soya beans (Glycine max L.) and one varietyeach of maize (Zea mays L.) and vetch (Vicia sativa L.) wereused. The seeds germinated when they had increased in size bya certain amount, if germination is taken as the time when theradicle first appears. The rate at which roots and shoots extendalso depend on soil water content. Glycine max L., Vicia sativa L., Zea mays L., Soya bean vetch, maize, seed germination, root extension, lateral root initiation, neutron radiography     

Dihydroquercetin protects barley seeds against mold and increases seedling adaptive potential under soil flooding

Barley (Hordeum vulgare L.) seeds were soaked in aqueous 10⁻⁴ M dihydroquercetin (DHQ) to examine its influence on seed germination and further growth of seedlings under optimal soil watering and flooding conditions. The adaptive potential of the plants was estimated by the content of thiobarbituric acid reactive substances (TBARs) and the activity of ascorbate peroxidase (AsP). High-grade seeds were germinated evenly under (−DHQ)- and (+DHQ)-treatments. Low-grade seeds soaked in DHQ, showed no mold and twofold germination rate in comparison with the same seeds soaked in water. The seedlings grown from the similarly germinated seeds did not differ from each other in the shoot growth, independent of the DHQ-pretreatment. The root growth was higher in DHQ-pretreated plants. Soil flooding suppressed the shoot and root growth rates in non-pretreated and DHQ-pretreated plants, however TBARs content was lower in the roots and leaves of (+DHQ)-seedlings as compared to the (−DHQ)-ones. The activity of AsP increased more significantly in the (+DHQ)-plants. The ratio between TBARs content and the AsP activity was lower in the leaves of (+DHQ)-plants both under optimal soil conditions and flooding. Thus, the treatment of low-grade barley seeds with DHQ protects the seeds against mold and increases adaptive potential of the seedlings.     

Primary and Lateral Root Development of Dark- and Light-Grown Cotton Seedlings under Salinity Stress*

Primary root growth dynamics and lateral root development of dark- and light grown cotton seedlings (Gossypium hirsutum L., cv. Acala SJ-2) were studied under control and salinity stress conditions. The seedlings were grown by two methods: A) in paper-lined, vermiculite-filled beakers with the plants growing between the paper and the glass wall (Gladish and Rost, 1993), and B) in hydroponics after germination and initial growth in germination paper rolls saturated with the treatment solutions (Kent and Läuchli, 1985). After germination, daily primary root elongation rate gradually incrased to a maximum, then gradually declined to close to zero for dark-grown seedlings, or to sustained rates of about 10 mm per day for light-grown control plants. Salinity stress delayed primary root growth and reduced peak elongation rates, without changing the general primary root growth pattern. These results suggest that salinity changed the time-scale, but did not modify the normal developmental sequence. Lateral root growth was more inhibited by salinity than primary root growth. In addition, elongation of lateral roots was more inhibited by salinity than their initiation and emergence. Light exposure of the shoot favored both sustained primary root growth from 7 days after planting, and lateral root emergence and growth. Salinity effects were more severe on seedlings germinated and grown in hydroponics (method B) than on vermiculite-grown plants (method A). These results emphasize the importance of growing conditions for the NaCl-induced effects on cotton root development. In addition, the differential effects of salinity on primary and lateral roots became evident, pointing to diverse control mechanisms for the development of these root types.     

A comparison of root architecture and shoot morphology between naturally regenerated and container-grown seedlings of Quercus ilex

We explored the different mechanisms developed by naturally regenerated seedlings of Quercus ilex L. (Holm oak) under Mediterranean conditions compared to container-seedlings commonly used in plantations. We examined the differences in root architecture (including topology and morphology) and shoot parameters. The results showed that there are many differences in the architecture of the root system as well as in the shoot morphology between the two types of seedlings. The naturally regenerated seedlings were smaller with regard to most of the shoot and root parameters, but they developed a longer taproot, only first order lateral roots, and presented a more herringbone-like root system compared to the container seedlings. Conversely, all types of container seedlings, were larger and had a more extended root system with many orders of lateral roots, while their taproot length was restricted within the container’s depth. The quotient log (α)/ log (μ) for all seedlings, showed a tendency to decrease with plant size. A strict herringbone root system with an elongated taproot may be the optimal root architecture for Quercus ilex L. seedlings in order to survive under Mediterranean conditions.     

STUDIES ON THE GERMINATION OF SEEDS OF THE ROOT PARASITES,ALECTRA VOGELII AND STRIGA GESNERIOIDES. I. ANATOMICAL CHANGES IN THE EMBRYOS

Anatomical changes in the radicle and shoot meristems of embryos of germinating seeds of the obligate root parasites, Alectra vogelii and Striga gesnerioides were studied. When germination of seeds was stimulated by cowpea (Vigna unguiculata) root exudate, growth occurred mainly in the radicular pole of embryos and minimally in the plumular pole, resulting in seedlings with elongated radicles. Maximum radicle elongations of about 3 mm in A. vogelii and 2 mm in S. gesnerioides were recorded during a period of 8 and 11 days, respectively. Analysis of the radicular tip during the course of seed germination revealed that the activity of the meristematic tissue progressively decreased until it completely disappeared. When germinated seeds were cultured on nutrient agar media, the radicle meristem of A. vogelii continued to grow producing a normal root with a root cap. On the other hand, the radicles of cultured S. gesnerioides seeds elongated only slightly before meristematic activity ceased. During continued growth of seedlings of both species on agar media, lateral roots whose tips had typical angiosperm root topography, were initiated from the radicle.     

Light Quality Regulates Lateral Root Development in Tobacco Seedlings by Shifting Auxin Distributions

Light is an important environmental regulator of diverse growth and developmental processes in plants. However, the mechanisms by which light quality regulates root growth are poorly understood. We analyzed lateral root (LR) growth of tobacco seedlings in response to three kinds of light qualities (red, white, and blue). Primary (1°) LR number and secondary (2°) LR density were elevated under red light (on days 9 and 12 of treatment) in comparison with white and blue lights. Higher IAA concentrations measured in roots and lower in leaves of plants treated with red light suggest that red light accelerated auxin transport from the leaves to roots (in comparison with other light qualities). Corroborative evidence for this suggestion was provided by elevated DR5::GUS expression levels at the shoot/root junction and in the 2° LR region. Applications of N-1-naphthylphthalamic acid (NPA) to red light-treated seedlings reduced both 1° LR number and 2° LR density to levels similar to those measured under white light; DR5::GUS expression levels were also similar between these light qualities after NPA application. Results were similar following exogenous auxin (NAA) application to blue light-treated seedlings. Direct [³H]IAA transport measurement indicated that the polar auxin transport from shoot to root was increased by red light. Red light promoted PIN3 expression levels and blue light reduced PIN1, 3–4 expression levels in the shoot/root junction and in the root, indicating that these genes play key roles in auxin transport regulation by red and blue lights. Overall, our findings suggest that three kinds of light qualities regulate LR formation in tobacco seedlings through modification of auxin polar transport.     

Seed germination and seedling growth of Zostera marina L. (eelgrass) in the chesapeake bay

Seed germination and seedling growth of Zostera marina L. were monitored in the Chesapeake Bay in 1979 and 1980. Harvested seeds were placed in small acrylic tubes at several sites representing the salinity range of Z. marina distribution. Seed germination was observed first in late September and continued through May, with peaks in the fall and spring. The majority of seeds that germinated (66%) did so between December and March when water temperatures ranged from 0–10°C. There was no correlation between sites (different salinity regimes) and frequency of germination rates, indicating that salinity was not a major factor in the germination process in this study. Additional information on seed germination was available for seeds collected in 1977 and 1980 and subsequently monitored for germination at only one site. These data were similar to germination frequency recorded in 1979–1980.Seedling growth was measured from individuals collected from an existing Zostera marina bed. Seedlings were collected from November through May, at which time we could no longer distinguish seedlings from existing vegetative stock. Growth was characterized by the increased length of the primary shoot, number of leaves per shoot and numbers of shoots per plant. Seedling growth was slow during the winter months (water temperature ? 10°C) but rapidly increased in the spring (temperatures > 10°C). The size range of the harvested seedlings indicated that seed germination in the field probably occurred from October through April, corroborating evidence from the seed germination experiments.     

Effects of drought stress on seed germination of ornamental sunflowers

The response to drought stress on germination was investigated on three hybrids of ornamental sunflower, ‘Hadar’, ‘Pazit’, and ‘Zohar’. Different levels of water potential [Ψ: 0.0 (control), ?0.15, ?0.30, ?0.45, ?0.60, ?0.75, and ?0.90 MPa] were adopted using polyethylene glycol-6000 (PEG6000) at four germination temperatures (15, 20, 25, and 30 °C). Final germination percentage, mean germination time, germination index, germination rate index, and germination stress tolerance index were used to evaluate the genotype response to PEG-induced water stress. Shoot and root length and fresh and dry weight were measured on seeds germinated at 20 °C under the different levels of water potentials. During germination, the three ornamental sunflowers showed to be more sensitive to suboptimal temperature than to supraoptimal. Decreasing water potential of imbibition solution progressively inhibited and delayed seed germination. Among cultivars, ‘Hadar’ and ‘Pazit’ performed better at temperature lower than 30 °C. ‘Zohar’ showed a lower sensitivity to PEG-induced water stress at all temperature conditions. Water stress during seed germination depressed the following seedling growth under favourable conditions. As a result, shoot and root length and fresh and dry weight was significantly lower in seedlings from seed germinated at ψ ≤ 0.45 MPa.     

Is it possible to manipulate root anchorage in young trees?

The optimal root system architecture for increased tree anchorage has not yet been determined and in particular, the role of the tap root remains elusive. In Maritime pine (Pinus pinaster Ait.), tap roots may play an important role in anchoring young trees, but in adult trees, their growth is often impeded by the presence of a hard pan layer in the soil and the tap root becomes a minor component of tree anchorage. To understand better the role of the tap root in young trees, we grew cuttings (no tap root present) and seedlings where the tap root had (?) or had not (+) been pruned, in the field for 7 years. The force (F) necessary to deflect the stem sideways was then measured and divided by stem cross-sectional area (CSA), giving a parameter analogous to stress during bending. Root systems were extracted and root architecture and wood mechanical properties (density and longitudinal modulus of elasticity, E _L ) determined. In seedlings (?) tap roots, new roots had regenerated where the tap root had been pruned, whereas in cuttings, one or two lateral roots had grown downwards and acted as tap roots. Cuttings had significantly less lateral roots than the other treatments, but those near the soil surface were 14% and 23% thicker than plants (+) and (?) tap roots, respectively. Cuttings were smaller than seedlings, but were not relatively less resistant to stem deflection, probably because the thicker lateral roots compensated for their lower number. Apart from stem volume which was greater in trees (+) tap roots, no significant differences with regard to size or any root system variable were found in plants (?) or (+) tap roots. In all treatments, lateral roots were structurally reinforced through extra growth along the direction of the prevailing wind, which also improved tap root anchorage. Predictors of log F/CSA differed depending on treatment: in trees (?) tap roots, a combination of the predictors stem taper and %volume allocated to deep roots was highly regressed with log F/CSA (R ² = 0.83), unlike plants (+) tap roots where the combined predictors of lateral root number and root depth were best regressed with log F/CSA (R ² = 0.80). In cuttings, no clear relationships between log F/CSA and any parameter could be found. Wood density and E _L did not differ between roots, but did diminish with increasing distance from the stem in lateral roots. E _L was significantly lower in lateral roots from cuttings. Results showed that nursery techniques influence plant development but that the architectural pattern of Maritime pine root systems is stable, developing a sinker root system even when grown from cuttings. Anchorage is affected but the consequences for the long-term are still not known. Numerical modelling may be the only viable method to investigate the function that each root plays in adult tree anchorage.