Esters hydroxycinnamiques et induction photopériodique florale in vitro chez Cichorium intybus

Hydroxycinnamic acid esters and photoperiodic flowering induction in vitro of Cichorium intybus.
In root tissues of Cichorium intybus L. cv. Witloof cultured in vitro, the photoinductive period occurs between the 8th and the 16th day after the start of the culture. The promotive light conditions are either long days (16 h photoperiod) or daily cycles of 9 h white + 15 h red light applied during the photoinductive period in an otherwise short day treatment. During the photoinductive period and under suitable irradiation conditions, the endogenous hydroxycinnamic acid esters (especially chlorogenic acid) show a regular development as from the 8th day, reaching a maximal level by the 16th day. The amount of these molecules then decreases rapidly, preceding the external expression of floral induction. When we apply non-inductive conditions [short days (9 h) or daily cycles of 9 h white + 15 h (far-red + red) light supplied from the 8th to the 16th day in otherwise short day conditions], the metabolic changes indicate the same pattern during the inductive period as mentioned above. However, a fundamental difference exists between the inductive and non-inductive conditions, so that the production of hydroxycinnamic acid esters is particularly high towards the end of the second week of in vitro development in the induced state. This increase is not primarily due to increased photosynthetic activity in long day conditions, since it occurs both in the long day treatments and in the treatments with daily cycles of 9 h white + 15 h red light, thus revealing the morphogenetic action of light via phytochrome. This accumulation of hydroxycinnamic acid esters is correlated with floral induction, which appears to be ineffective unless there is a certain minimum amount of these molecules in the tissues.     

Effects of light flashes on the dark closure of Albizzia julibrissin pinnules

An electronic flash unit is used to deliver, at the beginning of a 10 min dark period and within a few ms, large doses of light to Albizzia julibrissin pinnules, to ascertain their effects on the rate of pinnule closing. In a series of alternating light flashes at 710 and 550 nm, the first 710 nm light flash significantly retards closing. A following light flash at 550 nm negates the far-red induced delay. The second 710 nm light flash delays closing less effectively than the first when given within 4 s after the green flash, but is just as effective when given after 30 s. The delay brought about by the second 710 nm light flash is again abolished by a light flash at 550 nm. A light flash at 660 nm has no effect on pinnule closing by itself and is also ineffective in reversing the far-red induced delay. A series of ten 710 nm light flashes becomes most effective in delaying closure when there is a dark interval of one min between flashes. The closing delay induced by a 710 nm light flash escapes reversal by a 550 nm light flash when the dark interval between the two flashes exceeds 2–3 min. A 750 nm light flash has no retarding effect on pinnule closing, but it becomes effective when preceded by a 660 nm or 550 nm light flash. The results obtained are suggested to be due to light absorbed by phytochrome and an unknown photoreceptor with green, far-red photoreversal property.     

Phénolamides et induction florale de Cichorium intybus dans différentes conditions de culture en serre ou in vitro

Phenolamides and floral induction of Cichorium intybus in different conditions of culture in glass-room or in vitro. Three complexes between phenols and amines (phenolamides) have been found in Cichorium intybus L., a plant with an absolute requirement of vernalisation followed by long days for flowering. Upon hydrolysis, these complexes (A, B and C) liberate aromatic amines whose exact identification is in progress, but which are closely related to dopamine, tyramine and serotonin, respectively. In a first series of experiments, phenolamides were studied in the buds of plants grown in the greenhouse under varying conditions. Only buds from plants which flower in long days contained large amounts of these compounds. Much smaller amounts were found in buds at the end of vernalisation (at 2–4°C) before long-day treatment as well as in buds kept in the vegetative state after vernalisation by being grown in short days (8 h light) or in total darkness. In a second series of experiments, phenolamides were studied in bud-forming calli induced in vitro on explants of tuberised root. After sixteen days of culture in continuous light, large quantities of phenolamide were found in the buds and calli of the upper part of the explant, while the lower part which never produces buds contained much less. Buds formed under continuous light produce inflorescences in approximately one month. Various other culture conditions make it possible to maintain the explants in the vegetative state. This can be obtained by short-day conditions, or otherwise under continuous illumination by decreasing the sugar or increasing the NAA levels in the medium. After 13 days of culture, the phenolamide levels were much lower under all of these conditions, than under conditions favourable to floral induction. Compound C is absent or present in trace amounts in vegetative buds. The significance of the differences observed between floral and vegetative buds is supported by the sensitivity of the analytical techniques used. The accumulation of phenolamides in tissues of Cichorium intybus appears to be closely linked to floral induction. Under continuous light it begins very early in young buds and even in the calli that bear these buds.     

Optical properties of Lactuca and Taraxacum seed and fruit coats: Their role as light filters

The optical properties of seed and fruit coats were examined from several varieties of light-sensitive achenes. Taraxacum vulgare L. and Lactuca sativa L. cv. Grand Rapids achenes with dark fruit coats and L. sativa cvs Huvudsallat and Issallat with white fruit coats were examined. Transmission spectra varied among the different achenes: white fruit coats of Lactuca acted as neutral density filters between 450 and 780 nm, whereas Taraxacum transmitted 2–36% in this region. The ribbed fruit coat structure greatly affected transmission so that at different locations in the same coat, transmission varied between 20 to 80% at 660 and 730 nm. Fruit coats of Grand Rapids lettuce and Taraxacum transmitted more far-red than red light with T₆₆₀/T₇₃₀ ratios of 0.8 and 0.4, respectively. The relationship between the optical properties of fruit coats and light-stimulated germination is discussed.     

The effects of SAN 9789 and light on phytochrome and the germination of lettuce seeds

Photoblastic seeds (akenes) of lettuce (Lactuca sativa (L.) cv. Grand Rapids) were treated with SAN 9789 [4-chloro-5-(methylamine)-2-a, a, a,-trifluoro-m-tolyl-3-(2H)-pyridasinone]. The seeds weere placed in Petri dishes on filter paper soaked with water or SAN solution. The treatment increased the germination in darkness from 17% for water to 78% for SAN treated seeds. An irradiation with 5 min red light gave a germination of 98% both in water and in SAN. In water the effect of red irradiation could be reversed with a short irradiation (8 min) of far red light (17% germination), while in SAN solution the far red reversibility was poor (92% germination). If the far red light was given repeatedly (5 min per h) it had a slightly larger effect. If given continuously for 24 hours, the germination in water was decreased to 0.3% and in SAN solution to 9%. Possible mechanisms for the SAN effect are discussed.     

Evolution des glucides intratissulaires au cours de la néoformation des bourgeons par des explantats racinaires de Cichorium intybus cultivés in vitro

Variation of intratissular carbohydrates during bud formation in root explants of Cichorium intybus cultivated in vitro .
During the cellular activation that begins with excision of root explants from Cichorium intybus L. var. Witloof cv. Zoom cultured in vitro, hydrolysis of fructose polymers, in particular of the polyfructosans (inulin) takes place. The products of degradation are used to cover the energetic needs connected with the increase of the mitotic activity. After day 2 the intracellular carbohydrates (sucrose and reducing sugars) develop differently according to further development of the explants. When growth of unorganized callus is favoured and organ formation inhibited by medium supplemented with auxin, fructose is accumulated; but under bud-forming conditions it is the amount of sucrose that increases. These differences were most notable between days 3 and 10 in culture, the period during which primordia occurred in the shoot-forming callus     

In vitro pollination of isolated ovules of Cichorium intybus L.

 A method for the in vitro pollination of chicory (Cichorium intybus L.) ovules was developed. The purpose was to avoid self-incompatibility after in vitro self-pollination of ovules isolated from flower buds before anthesis and from open flower heads. Seedlings were obtained at a low percentage (0.76%), and the results were explained in terms of pollen viability, pollen germination on the ovule, embryogenesis studies and ploidy analysis. Received: 9 April 1997 / Revision received: 23 July 1997 / Accepted: 6 September 1999     

Efficient in vitro propagation of Agave parrasana Berger

An efficient method for the in vitro propagation of Agave parrasana Berger, an important ornamental plant species native to the state of Coahuila, México, was developed. Proliferation of good quality shoots was achieved on agar-solidified basal MS medium supplemented with L₂ vitamins and 13.3 μM benzyladenine. Rooting was successful in the basal medium with no growth regulators; however, a light intensity of 100 μmol m^-2 s^-1 was found to promote better rooting than 25 μmol m^-2 s^-1. This revised version was published online in June 2006 with corrections to the Cover Date.     

An in vitro model for the study of hydroponic forcing in chicory (Cichorium intybus)

An in vitro model for studying the influence of different factors on chicon formation during hydroponic forcing has been developed. The shoot apex was isolated from the chicory roots and cultured on a gelled nutrient medium. This medium was considered as a replacement of the root. Small chicons (5 g) were produced. Water and, more importantly, sucrose availability had important influences on the outgrowth of the chicons. When sucrose was added to the medium the chicon-weight increased two-fold. On a medium with low agar concentration (0.3% (w/v)), heavier chicons were produced compared with a medium with agar at 1.2% (w/v). Browning of the pith tissue (= flowering stem) decreased with increased agar concentration. The results presented indicate that the in vitro system can be used as a research model to study chicon development in relation to root functioning and composition. This revised version was published online in June 2006 with corrections to the Cover Date.     

Determination of floral organ type in cultured Silene shoot apices

Shoot apices of the long day plant, Silene coeli-rosa , were cultured on a basal medium (+3% sucrose) in non-inductive short days (SD) following their excision from plants which had been exposed to long day (LD) treatments in order to examine the period for determination of each floral whorl. In response to the inductive LD treatments, the pattern of whorl formation in vitro reflected their normal appearance in Silene : sepals, stamens 1–5, petals, stamens 6–10 and carpels, although the number of apices initiating each whorl was lower in vitro compared with apices in vivo. However, supplementing the medium with 7 instead of 3% sucrose corrected this deficiency and, for the first time, resulted in apices initiating floral whorls in SD. The interval between the shortest treatment to result in whorl initiation in vitro, 4 LD (which also resulted in 50% flowering in vivo), and the treatment which gave 50% initiation of the corresponding whorl in vitro, was taken to be the period for determination of that whorl. The determination times on the 3% medium were: sepals (2 days), stamens 1–5 (3 days), petals (3 days), stamens 6–10 (4 days) and carpels (4 days); all of these periods shortened to about 1 day on the 7% medium. Tissue culture did not perturb the pattern of initiation of each whorl since apices excised and cultured from plants which had received 7 LD + 2 SD, exhibited each whorl over the same time scale as those of intact plants which received the same treatment. The data are consistent with a sequential determination and initiation of each whorl in the order that they appear normally in Silene . Synchronisation of cell division, as represented by peaks of the mitotic index and G₂/G₁ ratios on day 8 (7 LD + 2 SD), did not occur in vitro but the mitotic index did not descend to zero, further emphasising that tissue culture did not perturb the Silene apex.     

Photocontrol of somatic embryogenesis and polyamine content in Araujia sericifera petals

The effects of photoperiod and end-of-day phytochrome control on somatic embryogenesis and polyamine (PA) content in Araujia sericifera petals have been studied. Petals from immature flowers were cultured under 16- and 8-h photoperiods. Far red (FR), red (R) and FR followed by R light treatments were applied at the end of the photoperiods for three weeks. The number of somatic embryos, callus weight and the levels of free and bound PAs in the cultured petal explants were determined 40 days after the beginning of light treatments. Long day (LD) promoted somatic embryogenesis but did not have any significant effect on PA content. Short day (SD) reduced somatic embryogenesis and enhanced total PAs, mainly in the form of bound spermidine. End-of-day FR treatment increased PA content and inhibited somatic embryogensis under LD but had no significant effect under SD. This effect of FR on PA levels was cancelled by R and was independent of the presence of silver thiosulphate in the medium. End-of-day R treatment reduced the total PA content under SD. However, end-of-day R increased or reduced somatic embryogenesis under SD depending on the presence or absence of silver in the medium. The results suggest a photoperiodic control of somatic embryogenesis and PA content in A. sericifera. The effects of end-of-day R and FR treatments depend on the length of the photoperiod. This finding and the FR/R photoreversibility of end-of-day treatments indicate that phytochrome may be involved in both somatic embryogenesis and accumulation of PA.     

Number of floral organs in Circaeaster agrestis (Circaeasteraceae) and possible homeosis among floral organs

98.9% of 5092 flowers from 1041 individuals of Circaeaster agrestis have five floral organs, the formula is P₃A₁G₁ (73.13%), P₂A₂G₁ (25.59%), and P₂A₁G₂ (0.22%). Only 0.4% of the flowers have six floral organs and the formula is P₃A₁G₂ (20 flowers) or P₃A₂G₁ (one flower). All these flowers have one vascular bundle in the pedicel and were considered to be normal ones. There are 33 flowers (0.65%) with six or more floral organs and two vascular bundles in the pedicel and we found traces of fusion of different degree of two flowers into one. These flowers were considered as abnormal. Therefore the normal number of floral organs of C. agrestis is five and occasionally six, and the floral formulas are P₃A₁G₁ or P₂A₂G₁, sometimes P₂A₁G₂, and occasionally P₃A₁G₂ or P₃A₂G₁. A tepal in P₃A₁G₁ may be replaced by a stamen in P₂A₂G₁ or by a carpel in P₂A₁G₂ or in reverse. A carpel in P₃A₁G₂ may be replaced by a stamen in P₃A₂G₁ or in reverse. We hypothesize that there are two possibilities for the number of the floral organs to be five (six), the result of reduction from P₃A₂G₂, or there exists homeosis among floral organs.     

In vitro development of angiosperm floral buds and organs

In the last twenty-five years, young inflorescences, floral buds and individual floral organs of a number of species have been cultured in vitro. There is considerable variability in the requirement of plant growth regulators and nutritional factors for flower development of different species. This variability is compounded by the fact that the hormonal and nutritional requirements are different at various stages of organ and floral development. Experimental studies on normal and mutant flowers in vitro have provided insights into some of the regulatory processes in floral organogenesis. The potential use of the in vitro technique in elucidating the various mechanisms in flower development is stressed.     

光调控在植物组织培养中的应用研究进展

光调控是植物组织培养中一种有效的环境控制技术。该文对近年来国内外有关光调控在植物组织培养中的应用,即光照强度、光周期、光质对组培植物的生长发育、光合作用、愈伤组织诱导及其增殖与分化、器官和体细胞胚发生、生理特性及次生代谢物质等方面的影响研究进展进行了综述,为植物细胞工程提供参考。     

The adaptedness of the floral phenotype in a relict endemic, hawkmoth-pollinated violet. 1. Reproductive correlates of floral variation

This paper examines the relationship between quantitative variation in floral morphology (sizes of petals, spur and peduncle) and maternal reproductive success (seed production) in Viola cazorlensic (Violaceae), a narrowly endemic violet of south-eastern Spain pollinated by day-flying hawkmoths (Sphingidae). This plant is characterized by broad intraspecific variation in size and proportions of floral parts. Floral morphology does not influence significantly the probability of fruit set. Among flowers setting fruit, spur length and size of petals have no significant effect on seed production, but capsules from long- and short-peduncted flowers contain significantly more seeds than capsules from flowers with intermediate peduncles. Individual plants differ significantly in average floral characteristics. Plants with comparatively long and short peduncles tend to produce more seeds than those with intermediate ones, even after accounting statistically for individual differences in flower production. These findings are interpreted as evidence of disruptive selection on peduncle length during the study season. Floral variability in this species may be explained by the combined action of disruptive selection on peduncle length (the character most variable among individuals) and little, if any, stabilizing selection on spur length and size of petals     

Effects of medium components and light on callus induction, growth, and frond regeneration in Lemna gibba (Duckweed)

Summary Basal media, plant growth regulator type and concentration, sucrose, and light were examined for their effects on duckweed (Lemna gibba) frond proliferation, callus induction and growth, and frond regeneration. Murashige and Skoog medium proved best for callus induction and growth, while Schenk and Hildebrandt medium proved best for frond proliferation. The ability of auxin to induce callus was associated with the relative strength of the four auxins tested, with 20 or 50 μM 2,4-dichlorophenoxyacetic acid giving the highest frequency (10%) of fronds producing callus. Auxin combinations did not improve callus induction frequency. Auxin in combination with other plant growth regulators was needed for long-term callus growth; the two superior plant growth regulator combinations were 10 μM naphthaleneacetic acid, 10 μM gibberellic acid, and 2 μM benzyladenine with either 1 or 20 μM 2,4-dichlorophenoxyacetic acid. Three percent sucrose was best for callus induction and growth. Callus induction and growth required light. Callus that proliferated from each frond’s meristematic zone contained a mixture of dedifferentiated and somewhat organized cell masses. Continual callus selection was required to produce mostly dedifferentiated, slow-growing callus cell lines. Frond regeneration occurred on Schenk and Hildebrandt medium without plant growth regulators but was promoted by 1 μM benzyladenine. Callus maintained its ability to regenerate fronds for at least 10 mo. Regenerated fronds showed a slower growth rate than normal fronds and a low percentage of abnormal morphologies that reverted to normal after one or two subcultures.     

Thoughts on the possible role of phytochrome destruction in phytochrome controlled responses

Abstract. Fluence-response curves for low-energy phytochrome responses show two steps at red light exposures c . two orders of magnitude apart. This Feature of the fluence-response relationship can be interpreted as the consequence of the following processes:

• 1. 

  Induction of a photoresponse by a very low level of Pfr.
• 2. 

  Activation of a Pfr-destroying enzyme above a threshold level of [Pfr].
• 3. 

  Dependency of the rate of Pfr destruction on [Pfr] once the threshold level is reached.

In this way, adaptation of the phytochrome control system to a broad range of light doses could be realized.     

Blue light induced inhibition of seed germination: The necessity of the fruit coats for the blue light response

The seeds (achenes) of Laportea bulbifera require a chilling to break their dormancy and are negatively photoblastic. Their germination is inhibited by both continuous blue light and continuous or prolonged far-red radiation. The germination of de-coated seeds, prepared by removing the fruit coats, however, was strongly inhibited by continuous far-red, but not by continuous blue light. Photoreversible germination by a brief irradiation with red light occurred when the chilled seeds were exposed to prolonged far-red light. These results suggest that far-red light may regulate the germination of L. bulbifera seeds through the phytochrome system which exists in the regions other than fruit coats and that the blue light reaction may be governed by other photoreceptor system(s).