The relationship between the activities of the pentose phosphate pathway and glycolysis during early stages of floral induction in spinach

Summary A quantitative cytochemical study was made of fructokinase, glucokinase, and fructokinase (both PFK-ATP and PFK-PP+F-2:6-P) activities in shoot apices of 4-week old Spinacia oleracea. The rates of activity of these enzymes in the central zone of the shoot apex of plants kept on a short day regime were compared with those from plants transferred from a range of timing up to 24 h to a continuous light regime when floral induction occurred. A mechanism is suggested explaining how no measurable change in activities of the enzymes assayed could still account for the availability of adequate levels G-6-P as substrate for pentose pathway activity which is almost doubled early on in cells of the central zone of shoot apices induced to flower.     

Restricted cell/cell communication in the shoot apex ofSilene coeli-rosa during the transition to flowering is associated with a high mitotic index rather than with evocation

Summary Plants ofSilene coeli-rosa given 5 or more long days (LDs) flowered, even when the LDs were followed by 48 hours of darkness before their return to short days (SDs). The mitotic indices of shoot apices from induced plants shortly after induction were significantly higher than the indices of shoot apices from vegetative plants. Two major mitotic peaks were observed in the shoot apices of plants given 7 long days (LDs) on day 8. One coincided with that reported byFrancis andLyndon (1979).Cell to cell movement was tested in the shoot apices of vegetative and LD treated plants using probes with a molecular size of 749 daltons (fluorescein-hexaglycine) and 847 daltons (fluorescein-leucyl diglutamyl leucine). These probes showed some movement in the shoot apices of both short day (SD) and LD treated plants, but fluorescein-leucyl diglutamyl leucine was immobile in the induced apices of 7 LD plants on day 8 at time intervals which coincided with major mitotic activity in the shoot apex. Symplasmic restriction in the shoot apex was also observed in plants given 8 LDs (i.e., plants not returned to SDs on day 7).In plants that were placed in 48 hours of darkness after the 7 LD treatment or in plants given 5 LDs, there was no strong peak in the mitotic index, even though all these LD treatments resulted in 100% flowering. In such plants no symplasmic restriction was found in the shoot. Thus the symplasmic restriction on day 8 of 7 LD plants is associated with the high mitotic index, but neither of these phenomena is an essential part of the evocation process.Abbreviations F(Glu)₂ L-glutamylglutamic acid conjugated to fluorescein isothiocyanate isomer I (F-) - F(Gly)₆ F-hexaglycine - FLGGL F-leucyl-diglutamyl-leucine - F(PPG)₅ F-the pentamer (propyl-propyl glycine) - LD long day - LDs long days - SD short day - SDs short days     

The CUP-SHAPED COTYLEDON2 and 3 genes have a post-meristematic effect on Arabidopsis thaliana phyllotaxis

Background and Aims The arrangement of flowers in inflorescence shoots of Arabidopsis thaliana represents a regular spiral Fibonacci phyllotaxis. However, in the cuc2 cuc3 double mutant, flower pedicels are fused to the inflorescence stem, and phyllotaxis is aberrant in the mature shoot regions. This study examined the causes of this altered development, and in particular whether the mutant phenotype is a consequence of defects at the shoot apex, or whether post-meristematic events are involved.Methods The distribution of flower pedicels and vascular traces was examined in cross-sections of mature shoots; sequential replicas were used to investigate the phyllotaxis and geometry of shoot apices, and growth of the young stem surface. The expression pattern of CUC3 was analysed by examining its promoter activity.Key Results Phyllotaxis irregularity in the cuc2 cuc3 double mutant arises during the post-meristematic phase of shoot development. In particular, growth and cell divisions in nodes of the elongating stem are not restricted in the mutant, resulting in pedicel–stem fusion. On the other hand, phyllotaxis in the mutant shoot apex is nearly as regular as that of the wild type. Vascular phyllotaxis, generated almost simultaneously with the phyllotaxis at the apex, is also much more regular than pedicel phyllotaxis. The most apparent phenotype of the mutant apices is a higher number of contact parastichies. This phenotype is associated with increased meristem size, decreased angular width of primordia and a shorter plastochron. In addition, the appearance of a sharp and deep crease, a characteristic shape of the adaxial primordium boundary, is slightly delayed and reduced in the mutant shoot apices.Conclusions The cuc2 cuc3 double mutant displays irregular phyllotaxis in the mature shoot but not in the shoot apex, thus showing a post-meristematic effect of the mutations on phyllotaxis. The main cause of this effect is the formation of pedicel–stem fusions, leading to an alteration of the axial positioning of flowers. Phyllotaxis based on the position of vascular flower traces suggests an additional mechanism of post-meristematic phyllotaxis alteration. Higher density of flower primordia may be involved in the post-meristematic effect on phyllotaxis, whereas delayed crease formation may be involved in the fusion phenotype. Promoter activity of CUC3 is consistent with its post-meristematic role in phyllotaxis.     

Molecular cloning and expression analysis of spermidine synthase gene during sex reversal induced by Ethrel in cucumber (Cucumis sativus L.)

A full-length cDNA encoding spermidine synthase CsSPDS of 317 amino acids was isolated from cucumber. CsSPDS showed high similarity to SPDSs of other species, with the greatest similarity to SPDS of Coffea arabica (identity, 87%). Northern blot analysis revealed that CsSPDS was expressed predominantly in rapid growing tissues, such as roots, young leaves and flower buds. The response of CsSPDS to the sex reversal of three-leaf stage androecious cucumber seedlings by Ethrel treatment was also examined. Our results showed that the CsSPDS was up-regulated in shoot apex during sex reversal process, which correlated with increases of spermidine synthase activity and free spermidine (Spd) content, and a decrease of putrescine (Put) content in shoot apices.     

Some Aspects of Ontogeny of the Shoot Apices of Solanum melongena L. and Capsicum annuum L.

The ontogeny of the vegetative shoot apices of two importantvegetable plants, Solanum melongena L. and Capsicum annuum L.is described. Each shoot apex is studied at different stagesof seed germination. The relationship between time of germinationand (i) area of vacuoles in the cell, (ii) total area of thecell, (iii) area of the nucleus in the cell, and (iv) ratioof area of vacuoles in the cell to the total area of the cellin each apex is examined. The differentiation of cytohistologicalzonation in both apices is distinct only after one or two leavesare initiated and developed. At a certain stage heterogeneityin staining in the peripheral region of the shoot apex of S.melongena is found. The zonation in both plants differentiatesgradually in histological and cytological features. Vacuolationincreases or decreases in all the cells of the shoot apex orin the cells of a particular region of the shoot apex at differentstages of its ontogeny.     

The relationship between the activities of the pentose phosphate pathway and glycolysis during early stages of floral induction in spinach

A quantitative cytochemical study was made of fructokinase, glucokinase, and fructokinase (both PFK-ATP and PFK-PP + F-2:6-P) activities in shoot apices of 4-week old Spinacia oleracea. The rates of activity of these enzymes in the central zone of the shoot apex of plants kept on a short day regime were compared with those from plants transferred from a range of timing up to 24 h to a continuous light regime when floral induction occurred. A mechanism is suggested explaining how no measurable change in activities of the enzymes assayed could still account for the availability of adequate levels G-6-P as substrate for pentose pathway activity which is almost doubled early on in cells of the central zone of shoot apices induced to flower.     

The response of short day plantChenopodium rubrum L. to abscisic acid and gibberellic acid treatment applied at two levels of photoperiodic induction

Abscisic acid (ABA) (5 x 10^-4M and 5 x 10^-5M) and gibberellic acid (1 x 10^-4M) was applied to the plumula ofChenopodium plants with partly (one dark period) or completely (three dark periods) fulfilled photoperiodic requirements for flowering. Morphological and cytoogical criteria were used to investigate the time-course of the differentiation of the treated shoot apices. Both substances were ineffective in increasing the mitotic activity of the shoot apex at the suboptimal level of induction. The degree of branching was temporarily stimulated by ABA and GA treatment under these conditions. Moreover, GA caused the elongation of the shoot apex. With the completely induced plants ABA hastened flowering and the rise in branching was observed in all the treatment 48 h following the application of growth substances.     

Increased Pentose Phosphate Pathway Activity Linked to Floral Induction in Apices of Spinacia oleracea during Short Days

A quantitative cytochemical study of glucose-6-phosphate dehydrogenaseactivity as a marker of the pentose phosphate pathway (PPP)was made on shoot apices of Spinacia oleracea kept either continuouslyin short days for up to eight weeks or transferred for a singleperiod of 20 h to continuous light after between three and eightweeks in short days. By the time the apices kept in continuousshort days showed morphological changes relating to the floralstate, the PPP activity was already elevated. From four weeksonwards, the apices were more readily induced to the floralstate as evidenced by the increased PPP activity. In addition,the level of PPP activity achieved in the short-day apices asthey progressed to the floral state was as great as that observedin the apices induced to flower by a 20-h day. Spinacea oleracea, pentose phosphate pathway, shoot apex, cytochemistry, glucose-6-phosphate dehydrogenase, floral induction     

SLOW MOTION Is Required for Within-Plant Auxin Homeostasis and Normal Timing of Lateral Organ Initiation at the Shoot Meristem in Arabidopsis

The regular arrangement of leaves and flowers around a plant''s stem is a fascinating expression of biological pattern formation. Based on current models, the spacing of lateral shoot organs is determined by transient local auxin maxima generated by polar auxin transport, with existing primordia draining auxin from their vicinity to restrict organ formation close by. It is unclear whether this mechanism encodes not only spatial information but also temporal information about the plastochron (i.e., the interval between the formation of successive primordia). Here, we identify the Arabidopsis thaliana F-box protein SLOW MOTION (SLOMO) as being required for a normal plastochron. SLOMO interacts genetically with components of polar auxin transport, and mutant shoot apices contain less free auxin. However, this reduced auxin level at the shoot apex is not due to increased polar auxin transport down the stem, suggesting that it results from reduced synthesis. Independently reducing the free auxin level in plants causes a similar lengthening of the plastochron as seen in slomo mutants, suggesting that the reduced auxin level in slomo mutant shoot apices delays the establishment of the next auxin maximum. SLOMO acts independently of other plastochron regulators, such as ALTERED MERISTEM PROGRAM1 or KLUH/CYP78A5. We propose that SLOMO contributes to auxin homeostasis in the shoot meristem, thus ensuring a normal rate of the formation of auxin maxima and organ initiation.     

Changes in the Pattern of Cell Division in the Shoot and Root Meristems of Lolium perenne during the Transition from Vegetative to Floral Growth

For Lolium perenne cv. Cropper, a system which resulted in 100%flowering comprised 90 short days (SD) at 4 ?C (vernalization)and 30 SD at 18 ?C followed by 8 long days (LD). The mitoticindex and G1 and G2 percentages were measured in the shoot androot apices of plants following 2, 5 or 8 LD and in SD controlssampled at the beginning and end of induction. Identical measurementswere made in plants given 48 SD at 18 ?C followed by 2, 5 or8 LD; plants remained vegetative in response to this treatmentlacking vernalization. Significant increases in both mitoticindex and meristem size occurred in the shoot apex in LD followingthe vernalizing, but not the non-vernalizing, treatment. A clusterof mitoses in the apical dome of the shoot apex was unique tothe vernalized plants given 5 or 8 LD. However, an increasein root meristem size occurred regardless of vernalization,but a significant increase in the mitotic index was limitedto vernalized plants given 5 or 8 LD. Whilst the vernalization-LDtreatment resulted in an increase in the G2 percentage in theshoot apex following 2, 5 or 8 LD, no such alteration was observedin the root meristem. Thus, the changes to the cell cycle whichcorrelated with flowering were increased mitotic indices andG2 percentages in the shoot apex at each sampling time and increasedmitotic indices in the root apex following 5 and 8 LD. Key words: Cell division, flowering, Lolium perenne L.     

Cytokinin production by Asparagus shoot apex cultured in vitro

The cytokinin producing capacity of asparagus (Asparagus officinalis L.) shoot apex was examined by means of shoot apex culture in vitro, where adventitious roots were never formed. The cultured shoot apices continued to diffuse a small but constant amount of cytokinin into the medium throughout five passages of subculture. The cytokinin content in the apices at the end of the subculture was not different from that at the beginning of the subculture. These results indicate a production of cytokinin by the apices. However, the finding is not in conflict with the hypothesis that the root tip is the major source of cytokinin supply, because the root tip of asparagus produced more cytokinin than the shoot apex and the decline of shoot growth observed during the subculture was partially restored by an application of zeatin into the medium.     

Histochemical Study of Photoperiodic and Low Temperature Effects on Floral Induction of Spinacia oleracea

Shoot apices of Spinacia oleracea plants have been induced toflower either by: (a) subjecting leaves to 24 h long day, or(b) exposure to a short photoperiod but displaced by 8 h (displacedshort day) in the usual 24 h short-day cycle, or (c) exposureto low temperature (5 °C) during the dark period of thenormal short day. A quantitative cytochemical assay of pentosephosphate pathway activity during floral induction indicatesan approximate doubling of the rate of activity when comparedto that of vegetative apices (short day) (21 °C). Exposure to either low temperature, or a displaced short photoperiodstimulates pentose phosphate pathway activity in the shoot apexin a manner similar to that seen by long-day induction. Thischange in metabolic activity is accompanied by changes in theshape of the shoot apex which resembles that seen at an earlystage during floral induction. Spinacia oleracea, pentose phosphate pathway, shoot apex, glucose-6-phosphate dehydrogenase, floral induction, chilling, displaced short day     

Efficient shoot and root formation from cotton shoot apices

Successful shoot and root induction were obtained from shoot apices of two cotton (Gossypium hirsutum L.) genotypes, Nazilli 84S and Çukurova 1518, which are widely planted in Turkey. Plant tissue culture systems were established on Murashige and Skoog (MS) medium supplemented with various plant growth regulators using seven-day-old shoot apices as explants. The shoot apex size was of 2–3 mm; it contained the meristem and unexpanded leaves. Shoot apices were placed on MS plus vitamins and combinations of various plant hormones. The best regeneration responses were obtained for cv. Nazilli 84S (98%) on MS + 0.1 mg/l kinetin (KIN) + 1 g/l polyvinylpyrrolidone (PVP) and for Çukurova 1518 (94%) on MS + 0.1 mg/l KIN + 2 mg/l NAA + 1 g/l PVP. Including germination, all regeneration and rooting processes lasted only 5 weeks. The shoot apices of both genotypes developed successfully without intervening callus formation, and no significant differences between cultivars were found. All regenerated plants of both genotypes were phenotypically normal and set seeds. This shoot meristem-based rapid regeneration method can also be used in the cases of biolistic and Agrobacterium-mediated transformation.     

Interrelationships of Cotyledonary Shoots in Pisum sativum

Unequal two shoot systems of tall and dwarf varieties of Pisum sativum L. ev, Improved Pilot and Meteor, respectively, were used in experiments where manipulative treatments involving the removal of one or other or both of the shoot apices were carried out. These were combined with GA treatments. Twenty four hours after treatment, ¹⁴CO₂ was applied to the lowest expanded leaf on the larger shoot, and the distribution of radioactivity was determined after a further 24 hours. Removal ot the apex of the larger shoot reduced the level of translocation from the treated leaf, the effect being enhanced by the additional removal of the other apex. This (Heel was more marked in the tall than in the dwarf variety. In untreated control plants of both varieties there was very little transfer ot labelled material from the larger (dominant) shoot to the smaller (weaker) shoot. This pattern was not affected by the removal of the apex from the weaker shoot. In Improved Pilot, removal of the apex from the stronger shoot led to considerable transfer of radioactive material to the weaker shoot, GA treatment having little effect. However, in Meteor, transfer of radioactivity from the stronger to the weaker shoot. after removal of the apex from the stronger shoot, only occurred after the application of GA. Removal of both shoot apices again resulted in low levels of transfer of labelled material to the weaker shoot in both varieties. These results are consistent with the hypothesis that there is competition between the two shoots for photosynthates produced by the leaves and that treatments reducing the competitive ability of one shoot, reduce the level of nutrients received by that shoot and can result in transfer of materials from it to the other shoot. Continuation of such a situation will result in increased inequality and perhaps ultimately in the death of the weaker shoot.     

Growth correlations and rna synthesis in different parts of the shoot apical meristem ofChenopodium rubrunt L. induced to flowering

Uridine-³H incorporation and RNA concentration were investigated in different parts of the shoot apical meristem ofChenopodium rubrum using autoradiography and cytophotometry. A single inductive cycle was sufficient to bring about postinductive first events in the shoot apex but not for complete flower differentiation. The initial activation of RNA synthesis manifested itself in all zones of the apex. The first increase was more conspicuous in the peripheral than in the central zone. The indications of the first events in the apices after a single inductive cycle disappear prior to morphological reversal to the vegetative state. Induction by three short days led to rapid flower differentiation. The increase in RNA synthesis and concentration was most conspicuous in the central zone in this case. The ratio of RNA synthesis and content between bud and leaf primordia (B/L) also change in relation to photoperiodic induction. In vegetative plants the B/L ratio was low while after induction it increased. The shifts in activity of RNA synthesis observed in the shoot apical meristem are related to the changes in growth activity of the different parts of the apex. The growth ratios in the apices bear the character of growth correlations. The change in the growth correlations following photoperiodic induction together with the total activation of RNA synthesis are considered to represent one of the first events of the transition to the reproductive state.     

Localization of starch in shoot apices of vegetative and photoperiodically induced plants ofChenopodium rubrutn

Starch was determined by means of IKI reaction in shoot apices ofChenopodium rubrum plants induced to flowering by two short days and in non-induced plants. Small starch grains were already observed in the meristematic cells at an age of four days after sowing. Larger grains were found in the subapical region of the apex. Heterogeneity increases during further growth of the plants in induced, as well as in non-induced vegetative plants. Starch disappears from the cells potentially giving rise to axillary buds, while the number and size of starch grains increase in cells from which leaf primordia will be formed. This metabolic specifity of leaf and bud primordia is preserved during morphological differentiation and applies to vegetative, as well as to prefloral apices of photoperiodically induced plants. The amount of starch in the different regions of the apex is linked rather with organogenesis than with the quantitative growth in the apex.     

Specific arrangements of cortical microtubules are correlated with the architecture ofmeristems in shoot apices of angiosperms and gymnosperms

Arrangements of corticalmicrotubules (MTs) as seen in median longitudinal cryosections of shoot apices of several angiosperms and gymnosperms were studied by indirect immunofluorescence microscopy.Bryophyllum, Clethra, Helianthus, Houttuynia, Vinca (angiosperms), andPinus, Cedrus, Cedrus andGinkgo (gymnosperms) were examined. In all angiosperm apices collected during the growing season, MTs were mainly arranged anticlinally in the tunica, randomly in the corpus, and transversely in the rib meristem. This pattern of arrangements of MTs was further confirmed by electron microscopy inBryophyllum apices. In the apices of winter shoots MTs in the rib meristem were arranged randomly, indicating a seasonal change with respect to their arrangment. In all examined gymnosperm apices, populations of superficial cells showed both random and anticlinal arrangements of MTs, in contrast to those of angiosperm apices that consistently show anticlinally arranged MTs. In the shoot apices of both angiosperms and gymnosperms, cortical MTs were arranged perpendicularly to the directions of cell expansion. The significance of MTs in the maintrnance of the different architectures of shoot apices in angiosperms and gymnosperms is discussed.     

Evaluation of salt tolerance in cultivated and wild tomato species through in vitro shoot apex culture

The possibility of using in vitro shoot apex culture to evaluate salt tolerance of cultivated (Lycopersicon esculentum Mill.) and wild (Lycopersicon pennellii (Correll) D'Arcy) tomato species was determined and related to the response obtained by callus culture. Both apices and calluses were grown on media supplemented with 0, 35, 70, 105, 140, 175 and 210 mM NaCl, and growth and physiological traits were determined. Most apices of L. esculentum did not develop roots from low NaCl levels, whereas the apices of L. pennellii were able to develop roots at the different salt levels. This different degree of salt tolerance between L. esculentum and L. pennellii was not, however, clearly shown on the basis of the shoot growth of the plantlets. The callus response was similar to that shown by the rooting parameters, as callus growth in response to increased salinity was much greater in L. pennellii than in the tomato cultivar. K⁺decreased more and proline accumulated less with salinity in shoots of L. esculentum compared to L. pennellii, whereas the opposite response was obtained in calluses. The results obtained in this study suggest that rooting parameters are the most useful traits for rapid evaluation and screening of tomato species and segregating populations through in vitro shoot apex culture. This revised version was published online in June 2006 with corrections to the Cover Date.     

Dipeptides and Amino Acids as Nitrogen Sources for the Callus of Atropa belladonna

Morphactin ester applied as a bark band to stems of Monterey pine seedlings (Pinus radiata D. Don) at first stimulated and then inhibited the rate of stem elongation over a 20-day period. The inhibition was caused by a direct action of morphactin on the shoot rather than by indirect action on the root system. ¹⁴C labelled morphactin applied to the bark accumulated in the shoot apices and the amount accumulated could be related to the inhibition of growth. More than 70% of labelled material isolated from roots and shoot apices was in the form of a metabolite similar to the carboxylic acid derivative of the morphactin ester. A preparation of ¹⁴C labelled acid derivative applied to the bark was translocated to the shoot apex and to the roots and inhibited stem elongation.     

Leaf Determination in the Fern Osmunda cinnamomea--A Reinvestigation

Earlier studies of leaf determination in the fern Osmunda cinnamomeaL., using the technique of excision and sterile culture, indicatedthat the apices of incompletely determined leaf primordia areconverted into shoot apices after isolation. Subsequently otherworkers questioned this interpretation, suggesting that a primordiumis determined as a leaf shortly after its inception and thatany shoot which arises from it is an adaxial adventitious bud.Because of the importance of this distinction in the interpretationof determination, the phenomenon has been reinvestigated byscanning electron microscopy and histological methods. Sincefixation for electron microscopy was inconsistence, the workwas performed on the primordia using resin models formed indental plastic moulds. Our data, on explanted P3, P4, and P5leaf primodia, provide strong confirmation for the interpretationthat the leaf apex is indeed converted to a shoot apex in incompletelydetermined primodia after explantation. This new evidence formsthe basis for interpreting leaf determination in this experimentalsystem.Copyright 1993, 1999 Academic Press Leaf determination, in vitro, surface replica, scanning electron microscopy, Osmunda cinnamomea L., cinnamon fern