The release of the cotyledonary shoot ofpisum sativum from inhibition in relation to changes in the levels of endogenous auxins,cytokinins, and gibberellins

Both axillary buds belonging to the cotyledons (cotyledonary buds) start to grow on decapitated pea seedlings, but one of them (the dominant shoot) prevails in growth over the other (the inhibited shoot). If the dominant' cotyledonary shoot is removed, the inhibited shoot is released from inhibition and starts to grow. This release from inhibition of the inhibited cotyledonary shoot is accompanied within two hours from the removal of the dominant cotyledonary shoot by a marked increase in the level of endogenous cytokinin-like substances and by a decrease in the level of endogenous IAA. By contrast, a significant increase in IAA level and a decreasing trend in the level of cytokinin-like substances occur in the originally inhibited cotyledonary shoot between hour 4 and hour 48 after the release from inhibition of the inhibited cotyledonary shoot. The level of gibberellin-like substances in the cotyledonary shoot released from inhibition steadily increases from the beginning of the release.     

The effect of stratification on the endogenous levels of gibberellins and cytokinins in seeds of douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) and sugar pine (Pinus lambertiana Dougl.)

Abstract Seeds of Douglas-fir and sugar pine which had been exposed to various periods of moist chilling (stratification) were extracted and bioassayed for gibberellin and cytokinin activity. In Douglas-fir a peak of gibberellin activity increased progressively during stratification and was seven times higher after 7 weeks, but declined subsequently. No change in the level of cytokinin activity was observed during stratification. However, a slight increase (1.7-fold) occurred in stratified seeds placed at 22°C for 3 days. In sugar pine a high level of gibberellin activity, approximately 20-fold higher than in unstratified controls, was present in extracts of seeds which had been stratified for 8 weeks. After 10 weeks of stratification cytokinin levels had increased approximately 200-fold. Four cytokinin-like substances were detected in seeds. Three of the substances are similar in their chromatographic properties to zeatin glucoside, zeatin riboside and zeatin, respectively. Both gibberellin and cytokinin activity declined rapidly after reaching maximum levels. In seeds with the coats removed, germination was rapid and complete in the absence of stratification. Prior to radicle emergence in coatless seeds cytokinin activity increased to a maximum (approximately a 1-7-fold increase) within 24 h. Significant levels of gibberellin-like substances were present after 8 h. Gibberellin activity could not be detected in coatless seeds which were imbibed in AMO-1618, and germination was inhibited significantly.     

The role of cytokinins in root development of pea seedlings

Endogenous cytokinin-like activity was detected in pea seedlings usingthe soybean callus bioassay. Higher levels of activity were found in two-day-oldseedling roots and in the root free zone of four-day-old seedlings compared tothe lateral root zone of four-day-old seedlings. By day six, the levels ofendogenous cytokinin-like activity was greatly reduced in both the lateral rootzone and root free zone. Decapitation of the root tip disrupted the spatialorganization of the root. Lateral roots were subsequently found along the entirelength of the root rather than in a discreet lateral root zone. Application of10^–3 M dihydrozeatin to decapitated root tipsovercame the effect of root tip removal and restored the situation to what isnormally found in intact roots. There was little mobility of dihydrozeatin inthe root, with applied ³H-DHZ not moving from the root free zone,even 24 h after application.     

Changes in Phytohormone Status in Stems and Roots after Decapitation of Pea Seedlings

Experiments were performed on the first and second internodes and 4-cm-long apical segments of main roots of pea (Pisum sativum L.) seedlings, grown in the light and decapitated above the second node on the seventh day after seed germination. Endogenous phytohormones were measured by the enzyme-linked immunosorbent assay during three days after decapitation of seedlings. The IAA level in the internodes decreased 2–3 times on the second day after decapitation of seedlings while the cytokinin level increased 5–6 times for zeatin and zeatin riboside (Z and ZR) and 1.5–2 times for isopentenyl adenine and isopentenyl adenosine (IP and IPA). In contrast to internodes, the IP and IPA contents in the roots of decapitated seedlings did not change, but the levels of Z and ZR increased 1.5–2 times compared to intact plant roots. The IAA level in the apical region of root remained almost unchanged after the removal of shoot apex. It was concluded that the apical meristem of the main root is not the site of the cytokinin response to the auxin signal coming from the stem apex and that a slight accumulation of Z and ZR after decapitation is due to upper zones of the root. There was no difference in the content of gibberellin-like substances between the internodes of intact and decapitated seedlings. However, the content of gibberellins (GA) in the root tip decreased after decapitation of seedling, which suggests an essential role of apical bud in supplying the root with GA and/or intermediates for their biosynthesis.     

Germination, seedling morphology and establishment of Cotnbretum bauchiense Hutch. & Dalz. (Combretaceae)

ONYEKWELU, S. S. C, 1990. Germination, seedling morphology and establishment of Cotnbretum bauchiense Hutch. & Dalz. (Combretaceae). Cotnbretum bauchiense is a suffrutex with short, erect, usually herbaceous stems arising from a woody root stock. It appears in savanna soon after fire and (lowers within a few weeks. The fruits germinate in 5–6 days. The germination is cryptogeal. On germination the true radicle and the apparent radicle formed by fused cotyledon stalks push down into the soil, carrying the plumule with them. The cotyledon lamina and part of the fused cotyledon stalks remain above the soil. Both the apparent radicle and the true radicle produce roots. Below the soil at the joint of the apparent radicle and the true radicle the plumule produces 1–3 shoots which grow out to the surface of the soil. The underground portion of the shoot bears scale leaves, from which the plant regenerates when the aerial shoot is damaged by fire. This type of germination is an adaptation that ensures successful establishment in an environment that is subjected to fire.     

The effect of culture medium composition and incubation time on synthesis of gibberellin-like substances by bacteria isolated from the roots of pine seedlings (Pinus silvestris L.)

It was found that synthesis of gibberellin-like substances by ten strains of Coryneform bacteria isolated from the roots of pine seedlings depended on both the composition of the medium and incubation time. More of these substances were produced in mineral medium with glucose in complex medium with casamino acids and yeast extract. Most gibberellin-like substances were found in 7 or 14-day old cultures. Culture supernatant fluids of most of the bacteria tested contained several gibberellin-like substances which on chromatograms run with the solvent system benzene, acetic acid (10:3, v/v) were located at Rf 0.0-0.3; 0.4-0.6 and 0.8-1.0.     

Starch metabolism in poplar wood ray cells during spring mobilization and summer deposition

Treatment with cyclohexylamine, an inhibitor of spermidine synthase, accelerated radicle emergence in chick-pea ( Cicer arietinum L. cv. Castellana) seeds. Stimulation in the growth of embryonic axis was correlated with: (a) a rise in putrescine (Put) and cadaverine (Cad); (b) a decrease in spermidine (Spd) and spermine (Spm), and (c) a concomitant acceleration of the transformation from S-adenosylmethionine (SAM) into ethylene with increases in the levels of l-aminocyclopropane-l-carboxylic acid (ACC), l-(malonylamino)cyclopropane-l-carboxylic acid (mACC) and ethylene and ACC synthase and ethylene-forming enzyme activities. Cyclohexylamine also stimulated the mitotic index in both apical and subapical zones of the radicle and the apical zone of the plumule. The sectional distribution of the ethylene pathway and polyamine content was studied in embryonic axes of seeds germinated for 65 h. Each axis was divided into 5 sections: radicle meristem, elongation zone, differentiation zone, hypocotyl and plumule. In the presence of cyclohexylamine, ACC synthase and ACC were strongly stimulated in both the differentiation and hypocotyl zones, whereas the mACC was stimulated in all sections of the embryonic axis. With respect to ethylene-forming enzyme activity and ethylene production, the hypocotyl and the zones of elongation and differentiation were affected most by cyclohexylamine. Cyclohexylamine also induced an accumulation of free Put and Spm in the differentiation, hypocotyl and radicle zones, whereas Put and Spm bound to small substances increased most in the hypocotyl and plumule. The Spd bound to small substances decreased in all sections in the presence of cyclohexylamine. With respect to polyamines bound to macromolecules, cyclohexylamine stimulated only the accumulation of Put since Spd and Spm were strongly inhibited in all sections.     

Effect of Salt Stress on Seed Germination and EmbryoGrowth of Spartina alterniflora (Gramineae)

In this paper , the effect of NaCl concentration ( 1× 104 - 6× 104 mg/L) on the seed germination of smooth cordgrass ( Spartina alterniflora) was investigated using 1/2 Hoagland as the basic culture solution. The results indicated that the coleoptile and embryonal axis grew before the radicle and plumule in the stage of germination . Salinity level had no influence on the seed germination percentage when the salt concentration didn′t exceed 3×104 mg/L. The optimal salinity level for seed germination and embryo growth was 1×104 mg/L . The inhibit effect of salt concentration on the growth of embryo′s different part was different, and the length of radicle and plumule decreased significantly with the increase of salt concentration. However , the lesser inhibition on the embryonal axis and coleoptile was observed under the higher salt concentration, and thus the seedlings out of soil were ensured and the species was quickly adapted to the environment of inter- tidal zone .     

In vitro germination and induction of direct somatic embryogenesis in 'Bottle Palm' [Hyophorbe lagenicaulis (L. Bailey) H.E. Moore], a critically endangered Mauritian palm

Hyophorbe lagenicaulis is a critically endangered palm of Mauritius. Zygotic embryos were isolated from seeds and germinated in vitro on MS salts and vitamins containing activated charcoal. When seedlings were pre-treated in vitro for 2 weeks in liquid medium containing 0.05 mg l^-1 paclobutrazol, 80% survived the transfer to soil. Three-week-old seedlings were sectioned longitudinally and partially embedded in medium that contained MS salts and vitamins, 30 g l^-1sucrose and 3 mg l^-1 2,4-dichlorophenoxyacetic acid. Somatic embryos were formed, in profusion, directly (without callus) from the haustorium, plumule and radicle. Direct regeneration is important for the conservation of endangered species, as fewer somaclonal variants are likely to arise than from indirect regeneration. When the haustorium, plumule and radicle of the longitudinally sectioned seedlings were separated, they formed more callus but fewer embryos. Plantlets derived from somatic embryos have not yet been successfully established in soil.     

Changes in protein profile of pigeonpea genotypes in response to NaCl and boron stress

Two pigeonpea [Cajanus cajan (L.) Millsp.] genotypes, a salt tolerant Manak and a salt sensitive ICPL 88039 were subjected to stress treatment of 3 mM boron, 60 mM NaCl and boron + NaCl at the seedling stage. Radicle and plumule proteins were analyzed by SDS-PAGE. Boron treatment increased 28.3 kDa proteins in plumule and 38.3 and 51.9 kDa proteins in radicle of Manak, however, there was no specific protein in ICPL 88039 either in plumule or in radicle. In NaCl treatment 95.6 kDa proteins appeared in plumule and 67.5 kDa proteins in radicle of Manak. Conversely content of some proteins decreased by boron treatment alone or in combination with NaCl although they were present in the controls. Thus, 54.3 kDa protein disappeared in ICPL 88039 plumule, 68.4 kDa in Manak radicle and 28.1 kDa in ICPL 88039 radicle.     

Thermal Hardening: A New Seed Vigor Enhancement Tool in Rice

In a laboratory study, indica and japonica rice (Oryza sativa L.) seeds were exposed to thermal hardening (heating followed by chilling followed by heating; chilling followed by heating followed by chilling; heating followed by chilling or chilling followed by heating). In indica rice, heating followed by chilling followed by heating resulted in decreased mean germination time, time to start germination, electrical conductivity of seed leachates, and time to 50% germination, as well as increased germination index, energy of germination, radicle and plumule length, root length, root/shoot ratio, root fresh and dry weight, radicle and plumule growth rate, and shoot fresh weight. In japonica rice, chilling followed by heating followed by chilling performed better than all other treatments, including control.     

亚精胺浸种对渗透胁迫下白三叶子萌发及淀粉代谢的影响

以‘拉丁诺’白三叶为材料,用0、10％、15％、20％（W/V）即的聚乙二醇（PEG-6000）溶液模拟干旱条件,研究亚精胺fSpd）浸种对渗透胁迫下白三叶种子萌发和淀粉代谢的影响。结果表明,在PEG渗透胁迫下,白三叶种子的发芽率、发芽势、发芽指数、胚芽及胚根鲜重和胚根长度均显著（P〈0．05）降低,淀粉水解为糖类的速率减慢;与蒸馏水浸种相比,0．05mmol．L-1 Spd浸种处理显著（P〈0．05）提高了在渗透胁迫条件下种子的发芽率、发芽势、发芽指数、胚芽及胚根鲜重、干重和胚根长度,同时大幅提高了α-淀粉酶、β-淀粉酶及（α＋β）．淀粉酶总活性,降低了淀粉含量,增加了还原糖和葡萄糖含量。说明Spd浸种提高了白三叶种子在渗透胁迫下的萌发能力和幼苗生长的环境适应性,这可能与增强种子体内淀粉酶活性,加速淀粉水解为还原糖和葡萄糖,为种子萌发和幼苗早期生长及时提供充足能量有关。     

Thermal Hardening： A New Seed Vigor Enhancement Tool in Rice

In a laboratory study, indica and japonica rice (Oryza sativa L.) seeds were exposed to thermal hardening (heating followed by chilling followed by heating; chilling followed by heating followed by chilling; heating followed by chilling or chilling followed by heating). In indica rice, heating followed by chilling followed by heating resulted in decreased mean germination time, time to start germination, electrical conductivity of seed leachates, and time to 50% germination, as well as increased germination index, energy of germination, radicle and plumule length, root length, root/shoot ratio, root fresh and dry weight, radicle and plumule growth rate, and shoot fresh weight. In japonica rice, chilling followed by heating followed by chilling performed better than all other treatments, including control.     

Production of auxins and gibberellin-like substances by mycorrhizal fungi,bacteria and actinomycetes isolated from soil and the mycorrhizosphere of pine (Pinus silvestris L.)

Summary Production of auxins and gibberellin-like substances by mycorrhizal fungi, bacteria and actinomycetes isolated from the mycorrhizosphere of Scots pine was studied. Chromatography and biossays were used.Most of the organisms required tryptophan for auxins production. The highest biological activity exhibited substances located at R_f 0.2–0.4.The organisms produced minute amounts of gibberellin-like substances which appeared at different R_f values. It was stated that auxins production is much more common among the root zone organisms of pine than the production of gibberellin-like substances.This research was carried out under problem MR.II.16 coordinated by the Institute of Dendrology, Polish Academy of Sciences.     

The HPLC profile of proteins of thermoprotection-acquired wheat (Triticum aestivum L.) seedlings

A pre-treatment of 40 °Cprovided thermoprotection to wheat seedlings against 43 °C, which was otherwise a lethal temperature. Due to temperature pretreatment, the rate of protein synthesis at 45 °C increased in both plumules and radicles. The HPLC profile of plumule and radicle proteins of thermoprotection-acquired seedlings was different from the plumules and radicles of non-treated seedlings.     

The effects of electric impulse on growth of Rhizophora mucronata seedlings (Rhizophorales: Rhizophoraceae)

Exposure of mangrove seedlings to electric current was studied at an intensity of 10, 20 and 30 mA to 0.5, 1, 3, 6, 12 and 24 h in different combinations, either to plumule or radicle or both parts of the propagules of Rhizophora mucronata Poir (sample size = 30). The electric current-treated propagules and untreated control were cultured hydroponically in sea water for 75 days, and root and shoot growth parameters measured. The root elongation was significantly influenced by duration of exposure, site of propagule treated and combined effects of current intensity x site of propagule treated. Leaf number and area were significantly influenced by intensity of electric current and duration of exposure; whereas shoot elongation was affected by the duration of exposure. The root number was significantly affected by duration of exposure. In general, the electric impulse improved the growth characteristics of shoot and root.     

The diffusion of gibberellins into agar and water during early germination of Pharbitis nil Choisy

Agar diffusion of imbibed seeds yielded significant amounts of diffusible Gibberellin-like substances. An analysis of the extractable and diffusible gibberellin-like substance, including an analysis of the remaining imbibition water of the seeds, indicated that a significant part of these gibberellin-like substances could be attributed to a net biosynthesis of these substances in the imbibing seeds. At the same time it was found that water diffusion yielded considerably more gibberellin-like activities than comparable agar diffusions i.e. 10 to 12 fold in general.Agar as well as water diffusion showed a temperature effect with regard to the yield of gibberellin-like substances particularly during the first 6 h of diffusion. The yield of these substances is lower at 10°C, and remains lower as shown with consecutive diffusions, in comparison with the yields at 20°C or 30°C.With both agar and water diffusion the sum of activities obtained with consecutive diffusions is always higher, often considerably higher, than equal periods of continuous diffusion which is probably due to inactivation and/or interference of inhibitory substances with the bioassay responses. Finally, water diffusates of both seeds and seedlings of the normal growing cv. Violet of Japanese morning glory contained considerably more gibberellin-like activities than those of the dwarf cv. Kidachi which indicated that normals synthesize more gibberellins than dwarfs.     

Endogenous gibberellin- and cytokinin-like substances in cultured shoot tissues of apple,Malus pumila cv. Jonathan,in relation to adventitious root formation

The frequencies of adventitious root formation in vitro of isolated shoots from bud cultures of apple (Malus pumila cv. Jonathan) after 1, 7 and 31 subcultures (weeks 5, 29 and 109 after the initial culture) were 5, 78 and 95% respectively. Endogenous gibberellin-like substances (GA) were extracted, chromatographed on SiO₂ partition columns, and assayed on dwarf rice (Oryza sativa cv. Tan-ginbozu). The levels of GA in shoots from the 1st, 7th and 31st subcultures were 40, 19 and 14 ng GA₃ eq./g dry weight of tissue, respectively, a trend which suggests an inverse relationship between endogenous GA level and rooting ability. This is consistent with the fact that applied GA₃ inhibits rooting in apple and many other species. The major peak of GA activity eluted coincidentally with GA₁/GA₃/GA₁₉. Endogenous cytokinin-like substances (CK) were chromatographed on paper and assayed with soybean hypocotyl sections. In contrast to the decrease in GA activity, CK activity increased 1.5–2.7 fold in the later subcultures (cytokinin activity per shoot, however, declined).     

Developmental morphology of seedling and shoot and phylogenetic relationship of Diplobryum koyamae (Podostemaceae)

We demonstrated that saltational evolution was recurrent in the body plans of seedlings of the aquatic angiosperm Podostemaceae, in contrast to other angiosperms with seedlings having almost common body plans. Diplobryum koyamae, transferred to the genus Hydrodiscus described in this paper, has long-floating shoots with an anchoring disk-like base and is rootless. Such a body plan is distinct from other members of Asian Podostemoideae comprising reduced or moderate shoots borne on the root. Here, our molecular phylogenetic analysis revealed that H. koyamae is sister to a crustose-rooted group of Hanseniella, Hydrobryum and Thawatchaia within Asian Podostemoideae. The germinating embryo was devoid of plumule and radicle, and comprised a single cotyledon and a short hypocotyl, which produced an adventitious shoot endogenously. The leaves are formed in the absence of the shoot apical meristem, accompanying the separation of lightly stained cells. Comparison with other species of Asian Podostemoideae having the plumule and the adventitious root in the seedling, along with their phylogenetic relationship, suggests that saltational evolution occurred in the seedling body plan of H. koyamae leading to the extraordinary adult body plan, as in the separate clade of Dalzellia, Indodalzellia, and Indotristicha of the subfamily Tristichoideae.     

Levels of gibberellin-like substances and their possible transport in developing dwarf and normal cultivars of pea (Pisum sativum L.)

The level of gibberellin-like substances was determined in the cotyledons and axis of developing seedlings of dwarf (Little Marvel) and normal (Tall Telephone) cultivars of pea (Pisum sativum L.). The effect of cotyledon removal with GA₃ application on growth was also examined. Greater levels of gibberellin-like substances were observed in the cotyledons of the normal cultivar than the dwarf. This was particularly evident in the cotyledons during the early stages of seedling growth. Subsequently there was a decline in GA levels in the cotyledons. This was coincidental with a rise in GA content in the axis with markedly greater levels in the normal than the dwarf cultivar. Decotyledonated dwarf and normal plants supplied with GA were much taller than the decotyledonated controls. This observation along with those of the gibberellin levels in the cotyledons and axis, provided circumstantial evidence that there may be translocation of gibberellins from the cotyledons to the axis.