The Metabolism of Zeatin and Zeatin Riboside by Soya Bean Callus

Five cell division inducing compounds were found in soya beancallus irrespective of whether it wes grown on a zeatin or zeatinriboside containing basal medium. In both cases the major metaboliteseems to be zeatin glucoside. The significance of this metabolicstep in plant tissue is discussed.     

Growth of Avocado Fruit Callus and Its Relation to Exogenous and Endogenous Cytokinins

Callus derived from avocado cotyledons and mesocarp was grown in tissue culture. Cytokinins in nutrient media were vital for the growth of mesocarp callus but not for the growth of cotyledonous callus. It was found that cotyledonous callus synthesized Cytokinins.     

Metabolism of N-(purin-6-yl)glycine by Soya Bean Callus

The growth of cytokinin-dependent soya bean callus has beenshown to be accelerated by adding N-(purin-6-yl)glycine to themedium. Two biologically active peaks were detected when thecallus was cultured with N-(purin-6-yl)glycine. These two peaksco-chromatographed with 6-(2, 3, 4-trihydroxy-3-methylbutylamino)purineand zeatin respectively. When ¹⁴C labelled N-(purin-6-yl)glycinewas applied to the callus, radioactivity was found with boththese compounds irrespective of whether or not the N-(purin-6-yl)glycinewas labelled in the side chain or in the 8-position of the purinering. Small amounts of zeatin appear to be produced from N-(purin-6-yl)glycinewhich could explain why this compound stimulates the divisionof soya bean callus. N-(purin-6-yl)glycine, soya bean callus, metabolism, radioactivity, cytokinins     

Regulation of Tracheary Element Differentiation by Exogenous L-Methionine in Callus of Soya Bean Cultivars

The relationship between the induction of tracheary elementdifferentiation and exogenous L-methionine was examined in agar-growncultures of soya bean callus initiated from Glycine max L. ‘Wayne’and ‘Clark 63’. Although Wayne is a normal cultivarsoya bean, seedlings of Clark 63 exhibit abnormal growth at25 °C due to exessive ethylene biosynthesis at this temperature.Wayne callus showed increased xylogenesis in the presence ofexogenous L-methionine (3.7 µg 1^–1) in comparisonto IAA–KN controls at both 20 and 25 °C. Clark 63callus produced greater numbers of tracheary elements in responseto exogenous L-methionine only at 25 °C. The induction ofxylem differentiation was independent of the maintenance temperatureof the stock cultures of both cultivars. Xylogenesis initiatedbyan IAA–KN medium was inhibited by the addition of AgNO₃(20 mg 1^–1) to the extent of 76.5 per cent in cv. Wayneand 6 per cent in cv. Clark 63. The inhibitory effect was partiallyreversed by the addition of L-methionine (3.7 µg 1^–1)to the IAA–KN–AgNO₂ medium. These data support thehypothesis that xylogenesis in vitro involves auxin, cytokininand ethylene. differentiation, xylogenesis, L-methionine, ethylene, Glycine max L., soya bean, callus culture, auxin, kinetin     

The Effect of Photoperiod on Levels of Endogenous Cytokinins in Xanthium strumarium

The cytokinin content of Xanthium strumarium L. plants decreased markedly when they were exposed to short days (SD). There was a significant decrease in the content of the butanol-soluble cytokinins of the mature leaves after only 5 SD cycles, and after 10 SD there was no significant cytokinin activity in butanol extracts; the changes in the young leaves were less marked. Most of the cytokinin activity in mature leaves appears to be present in the aqueous fraction, whereas in young leaves most activity occurs in the butanol-soluble fraction. SD treated plants produced less root exudate than LD plants, but there were no significant differences in the amounts of cytokinin in the root exudates from LD and SD plants collected over an equivalent time period. The cytokinin levels of SD-induced leaves remained low even when transferred back to LD. The observed differences in cytokinin levels did not appear to be the result of photosynthetic differences. Exposure of detached leaves to LD or SD did not result in differences in cytokinin content. It is not clear whether the observed changes in cytokinin levels in the leaves under SD are involved in the flowering response, but they may be causally related to a reduced chlorophyll content observed in SD-induced leaves.     

Dynamics of Endogenous IAA and Cytokinins during the Growth Cycle of Soybean Crown Gall and Untransformed Callus

The dynamics of the endogenous IAA and cytokinin levels duringthe growth cycle of two soybean crown gall lines (green andpale) induced by a nop⁺ Agrobacterium tumefaciens C58 strain,were compared with an untransformed soybean callus line. In both transformed tumor lines maximum cytokinin (essentiallyglucosyl-trans-zeatin) levels were attained in the beginningof the exponential growth phase, followed by a drastic decreasejust before the stationary phase was reached. Quantitativelythe green tumor line showed a 2–3 times higher cytokinincontent compared with the pale line. In the untransformed soybeancallus hardly any significant levels of cytokinin could be detected. Analysis of endogenous IAA levels showed no difference betweenthe two tumor lines and the untransformed callus tissue, allshowing a low and constant level throughout the entire growthcycle. The relevance of the endogenous accumulation of phytohormonesin relation to the hormone autotrophic growth of transformedsoybean tissue is discussed. ³ Senior Research Associate Nationaal Fonds WetenschappelijkOnderzoek (N.F.W.O.). ⁴ Recipient of an Instituut voor Wetenschappelijk Onderzoekin Nijverheid en Landbouw (I.W.O.N.L.) grant.     

Effects of Nitrate Level on Nitrogen Metabolism in Winged Bean and Soya Bean

The effects of high (15 mM) and low (0.75 mM) solution nitratelevels on nitrogen metabolism in three genotypes (IL 7A, IL13 and IL 21) of winged beans [Psophocarpus tetragonolobus (L.)DC.] and one genotype (Williams) of soya bean [Glycine max (L.)Merrill] were investigated. Plants were grown for 42 days ina greenhouse in solution culture prior to sampling. The 15 mM nitrate treatment resulted in greater growth of allplant parts except roots. Growth of soya beans was more responsiveto nitrate level than was growth of winged beans. The high nitratelevel inhibited nodulation in all plants. The IL 13 and IL 21winged bean genotypes had similar nitrogenase activity (acetylenereduction per plant) as the soya bean and IL 7A winged beangenotype had lower activity. However, the IL 13 winged beangenotype had higher nitrogenase activity (acetylene reductionper unit nodule mass) than the other three genotypes which allhad similar activity. The 15 mM solution nitrate level stimulatedleaf and root nitrate reductase (NR) activity for all plants.All winged bean genotypes had higher leaf NR activity and higherpercentage reduced- and nitrate-nitrogen contents of leavesand stems compared with soya beans. However, total protein (reducednitrogen) was greater in soya beans when sampled indicatingthat more nitrate had been metabolized by soya beans than bywinged beans during the 42-day growth period. Psophocarpus tetragonolobus (L.) DC., winged bean, Glycine max (L.) Merrill, Soya bean, nitrate reductase, nitrogen fixation, nitrogenase activity, nodulation     

The Role of Cytokinins and Ethylene in Bean Cotyledon Senescence. The Effect of Free Radicals

During ageing of bean (Phaseolus vulgaris L.) cotyledons in plants with modified life span the time-course of four cytokinins, ethylene, and the end products of free radical attack, lipofuscin-like pigments (LFP), were studied. UV irradiation shortened cotyledon life span, while epicotyl decapitation prolonged it. In controls, LFP increased at the senescence onset but at the end of life span it returned to the initial level. Ethylene increased more than 3-fold at the time of abscission. The content of individual cytokinins (zeatin, zeatin riboside, isopentenyl adenine, isopentenyl adenine riboside) varied differently during ageing but they did not decreased in any case under level observed in young cotyledons at the time of abscission. UV irradiation resulted in 14-fold increase in LFP concentration at the end. Ethylene increased 8-fold 2 h after irradiation. Individual cytokinins increased after UV irradiation to a different extent and time-course, nevertheless cotyledon life span was shortened. Decapitation induced LFP decrease. On day 13, LFP abruptly increased and than decreased and stayed lowered until abscission. Ethylene was maximum on day 24, at the time of abscission, it was above 200 % of control. Decapitation produced transient decrease in some cytokinins namely zeatin and isopentenyl adenine riboside.     

The Role of Cotyledons in Vegetative Growth and Nitrogen Assimilation by Soya Bean

The removal of both cotyledons from soya bean seedlings 10 daysafter sowing, when the primary leaves were unfolded, reducedtheir stem height, branching, leaf production and dry weightat flowering by a similar proportion whether they were nodulatedor nitrate-dependent. Nitrogen assimilation by the shoots ofnitrate-dependent plants was increased by the removal of onecotyledon and reduced by the removal of both cotyledons althoughthese effects were not significant. Both these treatments significantlyincreased the amount of nitrogen in the shoots of nodulatedplants at flowering, mainly by more than doubling the nitrogencontent of their leaves. In contrast, the proportion of thetotal plant nitrogen in the leaves of nitrate-dependent plantswas almost constant. These results suggest that the cotyledonsmarkedly inhibit nitrogen assimilation by nodulated plants butdo not appreciably affect nitrogen assimilation by plants dependentsolely on inorganic nitrogen for their nitrogen supply. Glycine mux (L) Merr., soya bean, cotyledons, nitrogen assimilation, growth     

Effect of Source of Nitrogen on the Growth of Fiskeby Soya Bean: the Carbon Economy of Whole Plants

Fiskeby V soya bean was grown from seed germination to seedmaturation with two contrasting patterns of nitrogen metabolism:either wholly dependent on dinitrogen fixation, or with an abundantsupply of nitrate nitrogen, but lacking root nodules. The carbonand nitrogen economies of the plants were assessed at frequentintervals by measurements of photosynthesis, shoot and rootrespiration, and organic and inorganic nitrogen contents. Plantsfixing atmospheric nitrogen assimilated only 25–30 percent as much nitrogen as equivalent plants given nitrate nitrogen:c. 40 per cent of the nitrogen of ‘nitrate’ plantswas assimilated after dinitrogen fixation had ceased in ‘nodulated’plants. The rates of photosynthesis and respiration of the shootsof soya bean were not markedly affected by source of nitrogen;in contrast, the roots of ‘nodulated’ plants respiredtwice as rapidly during intense dinitrogen fixation as thoseof ‘nitrate’ plants. The magnitude of this respiratoryburden was calculated to increase the daily whole-plant respiratory loss of assimilate by 10–15 per cent over thatof plants receiving abundant nitrate. It is concluded that ‘nodulated’plants grew more slowly than ‘nitrate’ plants inthese experiments for at least two reasons: firstly, the symbioticassociation fixed insufficient nitrogen for optimum growth and,secondly, the assimila tion of the nitrogen which was fixedin the root nodules was more energy-demanding in terms of assimilatethan that of plants which assimilated nitrogen by reducing nitratein their leaves.     

Determination by Gas Chromatography-Mass Spectrometry of [N(5)]Adenine Incorporation into Endogenous Cytokinins and the Effect of Tissue Age on Cytokinin Biosynthesis in Datura innoxia Crown Gall Tissue

In this study gas chromatographic-mass spectrometric techniques have been used to identify and quantify the metabolic incorporation of [¹⁵N₅]adenine into zeatin and its metabolites by 3-week-old Datura innoxia Mill, crown gall tissue. In a parallel study the levels of endogenous cytokinins were also determined by the stable isotope dilution technique using deuterium (²H)-labeled internal standards. Incorporation levels of the [¹⁵N₅]adenine after 8 hours of incubation, expressed as a percentage of the endogenous cytokinins, were as follows: zeatin (1.0%), zeatin riboside (1.5%), and zeatin riboside 5′-phosphate (10.2%). These results are consistent with those observed in complementary experiments using [U-¹⁴C]adenine, and support the proposal that the cytokinin biosynthesis occurs primarily at the nucleotide level. The effect of tissue age on cytokinin biosynthesis, determined by [U-¹⁴C]adenine incorporation into cytokinins by tissues at varying growth stages, indicated a steady increase with time reaching maximal synthesis at five weeks following subculture after which the level of ¹⁴C incorporation into cytokinins declined.     

Turnover of Storage Protein in Seeds of Soya Bean and Pea

We were interested in determining whether the low protein contentof pea seeds (Pisum sativum L.) as compared to soya bean seeds(Glycine max L. Merrill) might be due to faster degradationof the pea storage proteins during development of the seed.Pea and soya bean cotyledons were subjected to a ‘pulse-chase’experiment using [³H]glycine in in-vitro cultures. In peas,legumin had a half-life of 146 days, while vicilin had a half-lifeof 39 days. There was no measureable degradation of soya beanstorage proteins. Even with the pea storage proteins, the half-liveswere so much longer than the maturation time of seeds that degradationof storage proteins could not account for the lower proteincontent of peas as compared to soya beans. The validity of theseresults was indicated by the finding that non-storage proteinshad much shorter half-lives and that omission of a carbon ora nitrogen source greatly accelerated degradation. Labelledglycine was found to be a good probe for protein turnover studiesbecause it was very rapidly metabolized. Glycine max L. Merrill, soya bean, Pisum sativum, L. pea, protein turnover, storage proteins, legumin, vicilin     

细胞分裂素在果实及种子发育中的作用

就最近几年来细胞分裂素与果实及种子发育的研究进展作简要介绍。     

The Effect of Cytokinins on Vitrification in Melon and Carnation

The influence of exogenous cytokinin on vitrification of melonand carnation buds grown in vitro was assessed. In melon, theanatomical traits of vitrification were found to be similarto those previously described for carnation grown on a highconcentration of auxin. In carnation, the plantlets treatedwith high cytokinin concentrations became bushy but the anatomicalstructure remained normal. The relation of cytokinin and vitrificationis discussed. Cucumis melo L., Dianthus caryophyllus L., vitrified plantlets, habituation, cytokinin     

Endogenous Cytokinins and the Breaking of Dormancy and Apical Dominance in Potato Tubers

Cytokinins are present in all parts of potato tubers, and areequally distributed between the apical, lateral, and internodaltissue when dormant. However, the breaking of dormancy coincidedwith a rapid increase in the free base cytokinin levels in theapical buds and the tissue adjacent to it. These high levelsof cytokinin in the apical tissue were maintained while apicaldominance was displayed. Once apical dominance was overcomethe cytokinin levels in the lateral buds and the tissue adjacentto them were similar to the levels in the apical regions. Thepresent evidence suggests that cytokinin glucosides are transportedto the meristematic regions of the tubers where they are hydrolysedto their free bases. Amounts of free bases in excess of thoserequired for growth are apparently again converted to storageforms (particularly zeatin glucoside) in the meristematic regionsof the tubers and in the sprouts.     

Endogenous Gibberellins and Growth of Tobacco Callus Cultures

Tobacco callus grown under a range of conditions for different lengths of time contained various levels of gibberellin-like substances. Culture conditions, viz: light versus darkness and the quantity of cytokinin in the medium, affected the amount of gibberellins found in the tissue. These culture conditions were also important in controlling growth rate of the callus and modified the ability of the tissue to respond to exogenous gibberellins. Furthermore, substances which are known to inhibit gibberellin biosynthesis and also thought to block gibberellin action in some cases, were found to reduce the rate of growth. These data support the idea that endogenous gibberellins may be important in the control of the normal growth of tobacco cells in culture.