Abscisic Acid-Like Inhibitors and Cytokinins in the Developing Seeds of Dormant and Non-Dormant Varieties of Peanut (Arachis hypogea L.)

Abscisic acid-like inhibitors and cytokinin activities werestudied in the dormant and non-dormant peanut seeds during theirdevelopment from 20 to 50 d after anthesis. ABA-like inhibitorsincreased continuously from 20 to 50 d of seed development inthe dormant variety whereas in non-dormant seeds they increasedup to 40 d and then declined. Cytokinin-like compounds of boththe varieties increased from 20 to 30 d and then decreased upto maturity. Larger quantities of cytokinins were found at 30d of seed development in both the varieties. At the time ofmaturity dormant seeds contained greater amounts of ABA-likeinhibitors and less cytokinins than the non-dormant variety.     

Seed Dormancy in Acer: CHANGES IN ENDOGENOUS GERMINATION INHIBITORS, CYTOKININS, AND GIBBERELLINS DURING THE BREAKING OF DORMANCY IN ACER PSEUDOPLATANUS L.

Changes in germination inhibitors, cytokinins, and gibberellin-likesubstances during the breaking of coat-imposed dormancy of sycamoreseeds at low temperature suggest that dormancy may be controlledby the presence of endogenous germination inhibitors. Stratificationat 5°C led to a greater loss of neutral inhibitor(s) fromthe embryo than did incubation at 20°C. No marked differenceswere observed in the level of cytokinins and gibberellin-likesubstances between treatments. However, a gradual decrease inthe level of cytokinins was observed during stratification.Whether a cytokinin-inhibitor interaction is involved in thecontrol of dormancy of sycamore seeds, as was previously suggested,remains to be determined.     

Correlation of cytokinins and abscisic acid with monocarpic senescence in soybeans

The involvement of cytokinins and abscisic acid (ABA) in themonocarpic senescence (foliar yellowing following fruit development)of soybeans was examined. Foliar sprays of cytokinin (10^–4M zeatin or 10^–5 M benzyladenine), begun when the plantsfirst set fruit and repeated every other day, significantlydelayed, but did not prevent, monocarpic senescence. Foliarsprays of 10^–4 M ABA, applied in the same manner, significantlyhastened senescence of fruiting soybeans but apparently hadno effect on depodded plants. Leaf and stem material from pre-senescentand senescent plants was extracted, chromatographed, and bioassayedfor cytokinin-lilce activity (Amaranthus betacyanin productionassay) and ABA-like activity (oat coleoptile straight growthassay for inhibitors). ABA-like activity increased, and cytokinin-likeactivity decreased in shoot tissue before the plants began tosenesce. Cytokinin-like activity in the fruit also declinedduring this period. These results implicate a decrease in cytokininsand an increase in ABA-like inhibitors in the control of monocarpicsenescence of soybeans, but neither alone is causal. ¹ Supported in part by Research Grant 416-15-79 from the USDACooperative State Research Service under PL 89–106. ² Present address: Biology Dept., College of St. Benedict'sSt. Joseph, Minn. 56374, U.S.A. (Received February 4, 1978; )     

Physiology of Oil Seeds: II. Dormancy Release in Virginia-type Peanut Seeds by Plant Growth Regulators

Germination, ethylene production, and carbon dioxide production by dormant Virginia-type peanuts were determined during treatments with plant growth regulators. Kinetin, benzylaminopurine, and 2-chloroethylphosphonic acid induced extensive germination above the water controls. Benzylaminopurine and 2-chloroethylphosphonic acid increased the germination of the more dormant basal seeds to a larger extent above the controls than the less dormant apical seeds. Coumarin induced a slight stimulation of germination while abscisic acid, 2,4-dichlorophenoxyacetic acid, and succinic acid 2,2-dimethylhydrazide did not stimulate germination above the controls. In addition to stimulating germination, the cytokinins also stimulated ethylene production by the seeds. In the case of benzylaminopurine, where the more dormant basal seeds were stimulated to germinate above the control to a larger extent than the less dormant apical seeds, correspondingly more ethylene production was induced in the basal seeds. However, the opposite was true of kinetin for both germination and ethylene production. When germination was extensively stimulated by the cytokinins, maximal ethylene and carbon dioxide evolution occurred at 24 and 72 hours, respectively. Abscisic acid inhibited ethylene production and germinaton of the seeds while carbon dioxide evolution was comparatively high. The crucial physiological event for germination of dormant peanut seeds was enhancement of ethylene production by the seeds.     

Changes in Endogenous Cytokinins of Lettuce Seed during Germination

Using the soybean callus bioassay it has been shown that dormant lettuce seeds (Lactuca sativa L. cv. Grand Rapids) contain large amounts of water soluble cytokinins and small amounts of butanol soluble ones. When the seeds are irradiated with red light, or imbibed with 5 mg/1 gibberellic acid in the dark, the total cytokinin content of the seeds decreases, the level of water soluble cytokinins decreases, and the level of the butanol soluble cytokinins increases. Far-red light does not reverse this effect completely although cytokinin activity in the butanol extracts decreases following such irradiation. It is proposed that the interconversion of cytokinins initiated by red light, or gibberellic acid in the dark, is one of the primary events leading to radicle elongation in light-sensitive lettuce seed.     

Gibbercllins and Germination Inhibitors in Viable and Non-viable Seeds of Peanut (Arachis hypogea L.)

Gibberellins and germination inhibitors were studied in viableand non-viable seeds of peanut. More acidio and basic germinationinhibitors were present in non-viable than in viable seeds.Loss of viability is associated with the presence of inhibitorsand absence of gibberellin-like substances. Germination inhibitorsincluded compounds which co-chromatographed with abscisic acid.     

The influence of kinetin on the endogenous content of indoleacetic acid in swelling seeds of Phaseolus, Zea and Pinus and young plants of Phaseolus

Treatment of different plant materials, seeds of Phaseolus vulgaris, Zea mays and Pinus silvestris and young plants of Phaseolus, with kinetin increased the level of extractable IAA. For seeds this increase was most pronounced in bean seeds, which contained the lowest amount of endogenous IAA and cytokinins, and lower in maize seeds with high endogenous content of IAA and cytokinins. – For young bean plants the kinetin treatment significantly increased the extractable amounts of IAA from all parts of the plant, hypocotyls, cotyledons, epicotyls and primary leaves, when the cut plants were placed for 24 h in kinetin solution. For plants sprayed with kinetin solution only the primary leaves showed a significantly higher level of extractable IAA, which could be explained by the fact that the plants were growing very close together, so that the primary leaves received most of the kinetin during spraying.     

Relationships among cytokinins, ethylene and poly amines during the stratification-germination process in seeds of Acer saccharum

Relatively high levels of dihydrozeatin and trans-zeatin were detected in unstratified seeds of Acer saccharum Marsh. Both cytokinins increased substantially over the first 20 days of stratification at 5°C and then fell rapidly to values well below original levels by first germination on day 55. In seeds held at 20°C, a non-afterripening temperature, cytokinin levels remained constant for the first 10 days and then declined to their lowest levels by day 20. Levels of putrescine, spermidine and spermine in the radicles and cotyledons did not change during the full course of the afterripening process, but large increments were noted during radicle emergence. A large increase in ethylene production at germination suggests that competition for S-adenosyl-methionine by the ethylene and polyamine biosynthetic pathways did not inhibit synthesis of ethylene or polyamines during seedling emergence and establishment. In seeds stratified at 20°C, ethylene showed an exceptionally large peak early in the stratification period, but polyamine contents remained low throughout the test. The present results are consistent with the hypothesis that cytokinins play a significant role in overcoming the metabolic block present in dormant seeds. This conclusion is supported by data showing that high levels of cytokinins develop concurrently with the start of tissue differentiation and at the time when abscisic acid and phenolic inhibitors decline in stratifying seeds. Changes in ethylene and polyamine contents did not correlate with any events in the afterripening process; however, large increases in levels of these substances were closely associated with the germinative process and, in the case of polyamines, specifically with the start of cell division.     

Survey of plants for enterokinase inhibitors

A radioisotopic assay procedure was developed for the assay of enterokinase inhibitors. This assay was used to test a number of plant species for enterokinase inhibiting activity. Enterokinase inhibiting activity was found in 5 of the tissues surveyed. These included peanut seeds, leaves of two tomato varieties, and tubers of two potato cultivars. Peanut seeds contained protein enterokinase inhibitors that were separated by gel filtration on Sephadex G-50 after quantitative recovery by affinity chromatography on anhydrotrypsin-Sepharose. Enterokinase inhibiting activity was found to accumulate in tomato leaves as a result of wounding. It was concluded that some plants contain enterokinase inhibitors.     

Could plant hormones be the basis of maturation indices in Pinus radiata?

Maturation and phase change in woody plants are developmental processes at present poorly understood, especially from a physiological point of view. Maturation indices, such as phytohormones, could be used to understand more about the events that accompany the increase in the developmental state and to choose optimal conditions in order to improve forest programs and make them more profitable. In order to determine this putative index, the contents in abscisic acid-like substances (ABA-like), indole-3-acetic acid (IAA) and several cytokinins (Cks) were analysed in terminal and axillary buds of Pinus radiata trees with different developmental states to compare their phytohormonal status. An increase in the content of some zeatin-type (Z-type) Cks, and a decrease in isopentenyladenine-type (iP-type) Cks levels throughout maturation were shown both in terminal and axillary buds. This fact allowed us to consider the ratio iP-type/Z-type Cks, which decreased in parallel with the increasing developmental state of the tree, as a maturation index. According to the results, ABA-like compounds and IAA contents could not be considered as a maturation index in Pinus radiata, even though similar patterns were in fact found between the two kinds of buds.     

Somatic embryogenesis and organogenesis in peanut: The role of thidiazuron and N6-benzylaminopurine in the induction of plant morphogenesis

Intact peanut (Arachis hypogaea L.) seeds, incubated on media containing N⁶-benzylaminopurine (BAP) or thidiazuron (TDZ) exhibited de novo regeneration at the hypocotyledonary notch region. Regeneration was observed when seeds were cultured on either TDZ or BAP but the optimal level of media supplementation was 10 mol·L^–1 for TDZ and 50 mol–L^–1 for BAP. Light microscopic observations revealed that the regenerants induced by TDZ were somatic embryos while those induced by BAP were shoots. An alternative approach of exposing the seeds to TDZ was through vacuum infiltration followed by culture on basal media but BAP did not induce regeneration by this method. Although TDZ has often been classified as a synthetic cytokinin, our results clearly demonstrate that seedlings treated with TDZ undergo a different morphological route of development than that induced by purine cytokinins.     

Ethylene as a Component of the Emanations From Germinating Peanut Seeds and Its Effect on Dormant Virginia-type Seeds

The embryonic axes of Spanish-type peanut seeds that do not exhibit dormancy to any extent were found to produce ethylene during germination. Virginia-type peanut seeds of the extremely dormant variety NC-13 produced low levels of ethylene when imbibed but not germinating. Treatments that released dormancy of NC-13 peanut seeds resulted in increased ethylene production by the embryonic axis. The estimated internal concentration of ethylene in Virginia-type peanut seeds was 0.4 ppm at 24 hr of germination. Fumigation with an external concentration of 3.0 to 3.5 ppm for 6 hr was sufficient to break dormancy of Virginia-type peanut seeds. These results suggest that ethylene is associated with the germination processes of non-dormant seeds and participates in the breaking of seed dormancy of dormant peanut varieties.     

Isolation and expression analysis of LEA genes in peanut (<Emphasis Type="Italic">Arachis hypogaea</Emphasis> L.)

Late embryogenesis abundant (LEA) protein family is a large protein family that includes proteins accumulated at late stages of seed development or in vegetative tissues in response to drought, salinity, cold stress and exogenous application of abscisic acid. In order to isolate peanut genes, an expressed sequence tag (EST) sequencing project was carried out using a peanut seed cDNA library. From 6258 ESTs, 19 LEA-encoding genes were identified and could be classified into eight distinct groups. Expression of these genes in seeds at different developmental stages and in various peanut tissues was analysed by semi-quantitative RT-PCR. The results showed that expression levels of LEA genes were generally high in seeds. Some LEA protein genes were expressed at a high level in non-seed tissues such as root, stem, leaf, flower and gynophore. These results provided valuable information for the functional and regulatory studies on peanut LEA genes.     

Time course of the changes in the level of endogenous growth regulators during the stratification of the seeds of the ‘Panenské české’ apple

The time course of the changes in the level of endogenous growth regulators was followed during the stratification at 5 °C of the seeds of ‘Panenské ?eské’ apples. An increase in the endogenous gibberellin activity was found already during the first and the second week of stratification which is according to it decisive for the release of dormancy in the seeds, as it precedes with the anticipation of approximately two weeks the curve of the release of dormancy in the seeds. The rise in the level of endogenous cytokinins in the seeds is belated one to two weeks behind the rise in gibberellin activity in them and thus approximately coincides with the release of dormancy. The rise in auxin level occurs approximately 4 weeks after the increase in cytokinin level. The increase in auxin level, which is accompanied by an increase in inhibitions, is apparently not connected with the release of dormancy in the seeds during the stratification.     

Enhancement of ethylene biosynthesis and germination by cytokinins and 1-aminocyclopropane-l-carboxylic acid in Striga asiatica seeds

Germination of witchweed ( Striga asiatica [L.] Kuntze), an important parasitic weed on several poaceous crops, is stimulated by several synthetic and natural compounds. We investigated the role of ethylene biosynthesis and action in cytokinin-induced germination. Conditioned Striga seeds treated with distilled water, 1-aminocyclopro-pane-1-carboxylic acid (ACC) or the cytokinins thidiazuron (TDZ), trans zeatin (TZ), benzyladenine (BA) and kinetin (KIN) produced little ethylene. Treatments with cytokinin-ACC combinations enhanced ethylene production. The relative order of activity of the cytokinins in elicitation of the phytohormone was TDZ > TZ > BA > KIN. Germination in response to distilled water and ACC treatments was negligible. Induction of germination by cytokinins varied from low (0%) to moderate (52%). Seeds treated with cytokinin-ACC combinations displayed high rates of germination. The observed germination was positively correlated (γ= 0. 8 and 0. 9) with ethylene production. Germination was reduced by silver thiosulphate (STS) and CoCl₂, inhibitors of ethylene action and ACC oxidase, respectively. Aminoethoxyvi-nylglycine (AVG), an ACC-synthase inhibitor, reduced TDZ-induced Striga germination. However, the inhibitory effect of AVG was overcome by addition of ACC. The results are consistent with a model in which Striga germination and embryo growth are limited by low capacity of the seeds to oxidize ACC. The cytokinins promote ACC conversion into ethylene and consequent Striga germination by enhancing ACC oxidase activity and/or synthesis.     

Cytokinins in maturing and germinatingLupinus luteus L. seeds

³H-labelled zeatin riboside (ZR) was applied to pod walls of intactLupinus luteus L. plants. Metabolites present in mature, dry seeds were zeatin nucleotide (ZNT), zeatin riboside (ZR) and zeatin (Z), zeatin O-glucosides and lupinic acid (LA), and the corresponding dihydro-derivatives of the cytokinins listed. Endogenous cytokinins were rapidly metabolised in germinating seeds. In seeds labelled with [³H]ZR for 90 min following a 2 h period of imbibition in water, ZR was actively converted to ZNT and dihydro-ZNT but the prevailing CTK was Z in cotyledons and ZR in embryo axes (EA); later LA and dihydro-LA, and O-glucoside metabolites accumulated. When [³H] zeatin was introduced into imbibing seeds, it was converted to dihydro-ZNT, ZNT, dihydro-ZR, ZR and dihydro-Z; in EA of the Z-labelled seeds, dihydro-ZR and ZR were the main cytokinins. After incubation of the Z-labelled seeds for 6 h in water, the ratios of dihydro-ZNT: ZNT and dihydro-ZR: ZR were, respectively, 20: 1 and 3.4: 1 in EA, and 3.5: 1 and 1.4: 1 in cotyledons.     

顽拗性黄皮胚脱水过程中核酸、蛋白质代谢的变化

黄皮种子发育晚期,胚内核酸、蛋白质合成能力增强,而花生胚的核酸、蛋白质合成能力在发育晚期则呈下降趋势。黄皮胚的发育在达到生理成熟后维持着活跃的生理代谢并转入萌发状态;而花生胚的代谢活性逐步降低并转入生理静止状态。脱水处理引起生理成熟期黄皮胚核酸、蛋白质合成能力急剧下降,核酸水解酶活性增强。不同程度脱水的黄皮胚吸胀２４ｈ,核酸、蛋白质的合成能力随脱水程度的加深而降低;生物大分子代谢能力的变化是顽拗性