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     

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.     

Fissazione di C14O2 in Colture di Tessuti Vegetali « in Vitro »

Abstract

C¹⁴O₂ fixation in plant tissues « in vitro ». — In the present work it has been examinated the autotrophic and heterotrophic CO₂ fixation of explants of « Helianthus tuberosus » « in vitro » and the photosyntetic efficiency of leaves produced from buds of « in vitro » explants of « Cichorium intybus » compared with that of mature leaves from normal plants of the same species. From our results it is evident that « in vitro » explants of « Helianthus tuberosus », grown, in the light, are able to autotrophically incorporate C¹⁴O₂; the distribution of the radioactivity into the various fractions shows a large influence of the light on the neutral fraction containing sugars (50% of the total radioactivity). In the chlorophyllous explants the dark CO₂ fixation is obviously of heterotrophic type: 97% of the total radioactivity is incorporated in amine acids (43%) and the organic acids (53%); on the other hand in the dark grown explants the radioactivity is differently distributed between amino acids (59%) and organic acids (39%). Mature leaves from normal plants and leaves produced from buds of « in vitro » explants of « Cichorium intybus » incorporate the same quantity of C¹⁴O₂ when expressed per mg of chlorophyll; the different distribution of the radioactivity in the neutral and acid fractions could be explained in terms of a different utilization pathway of the photosynthates in the two tissues.     

The characteristics of light in floral induction in vitro of Cichorium intybus. The possible role of phytochrome

The action of light in the initiation of floral buds in vitro has been studied using monochromatic light qualities on root explants of a long day plant, Cichorium intybus L. cv. Witloof. Red light (660 nm, 0.30 W m^-2) promotes flowering, while far-red (730 nm, 0.31 W m^-2) and irradiation with combined red + far-red (0.20 + 0.41 W m^-2) have no effect. In short day conditions floral response can be obtained in two ways: 1) by interrupting the dark period with 5 brief irradiations of red light (0.45 W m^-2, 12 min) at regular intervals, although these are counteracted by far-red irradiations of equal intensity and duration; 2) by interrupting the long night with 5 h red light applied during the second third of the night, while at the beginning or at the end it is ineffective. Red light efficiency appears to depend on the photosynthetic activity of the tissues, so that flowering increases with increasing intensity of white light and is suppressed if no white light is supplied. The reproductive development is determined by the coordination of proper irradiation conditions with sufficient sensitivity of the perceiving meristematic cells. The period of highest sensitivity to environmental light conditions in the life cycle of a Cichorium root explant occurs between the 8th and the 16th day after the start of the culture. The data strongly suggest that phytochrome is involved in flower induction of Cichorium in vitro.     

Etude du métabolisme des acides nucléiques et des protéines au cours de l'initiation de racines adventives

Summary Changes in the metabolism of nucleic acids and proteins were followed during the initiation of adventitious roots of Cicer arietinum. During the first 24 h, several phases of activation of the metabolism were found. The use of inhibitors of RNA synthesis showed that an early phase of incorporation of precursors into RNA is essential for the initiation phenomena. This phase is more sensitive to inhibitors and activators (hormones) than all other phases. The rapid turn-over of RNA synthesized during the first 6 h and the inefficience of inhibitors after this time suggested that a stable (non-RNA) factor maintains the activation. This factor must be synthesized soon after the stimulus of cutting.     

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.     

The effect of cold, anoxia and ethylene on the flowering ability of buds of Cichorium intybus

The apical bud and the axillary buds of Witloof chicory ( Cichorium intybus L. cv. Tardive d'Anvers) remain in the vegetative state if they are left on the root and maintained at 18°C. Flowering occurs in long days of 16 h after a pretreatment of either 8 weeks at 3°C, 3 days in complete anoxia at 15°C, or 4 days in the presence of ethylene (1000 ppm) at 15°C. In contrast, the adventitious buds which spread out on the root after ablation of the collar flower in a photoperiod of 16 h without particular pretreatment. The grafting of apical buds onto roots after different treatments shows that cold and ethylene act on the root, whereas anoxia acts directly at the level of the bud. It seems that the inhibition of the flowering of preformed buds (apical and axillary) stems from the collar. A hypothesis is proposed to explain this inhibition and why it is broken by cold, anoxia and ethylene.     

Distribution of mRNAs of fibrinogen polypeptides and albumin in free and membrane-bound polyribosomes and induction of α-fetoprotein mRNA synthesis during liver regeneration after partial hepatectomy

To study the effect of regenerative response of the liver following partial hepatectomy on the synthesis of major plasma proteins (secretory proteins), we have determined the sequence contents and the distribution of albumin and fibrinogen polypeptide mRNAs in rat liver at intervals after partial hepatectomy and sham operation. Using a quantitative technique for the isolation of polyribosomes, we demonstrated that the distribution of RNA between free and membrane-bound polyribosomal fraction was unchanged in these experiments. There was no shift in the polyribosomal population to favor free polyribosomes after partial hepatectomy. However, there was a dramatic increase (5–6-fold) of the fibrinogen polypeptide mRNA concentration during the first 24 h after resection. In contrast, the albumin mRNA concentration decreased (2–3-fold). There were no α-fetoprotein mRNA sequences detectable in any liver RNA fraction in these experimental animals. In sham-operated rats with intact livers, similar changes of fibrinogen polypeptide and albumin mRNA concentrations as described in regenerating liver after partial hepatectomy, were observed. These results suggest that albumin and fibrinogen synthesis after partial hepatectomy is reciprocally regulated at the mRNA level and represents a nonspecific acute phase response to surgical trauma.     

Influence of light conditions on development, apoplastic peroxidase activities and peroxidase isoenzymes in chicory root explants

Cichorium intybus L. (cv. Bea) root explants grown in continuous light had a higher fresh weight, lignin and chlorophyll content than explants grown in darkness. Intermediary values were found when the light conditions were switched after 6 days. Peroxidase activities (EC 1.11.1.7) were measured in the apoplastic fluid with guaiacol, indoleacetic acid (IAA) and syringaldazine as substrates. There was an inverse relationship between specific IAA oxidase activity and explant growth. Specific syringaldazine oxidase activity (SSO) correlated with the lignin content. Analysis by isoelectric focusing (IEF) showed that the apoplastic fluid contained both cationic and anionic peroxidase isoforms, whose expression differed according to culture conditions. Isoform isoelectric point (pI) 7.0 was only detectable when the explants were cultured for 12 days in darkness. When explants were grown for the first 6 days in light, SSO and lignin content were high and the isoforms pI 4.0, 6.6, 7.6 and 8.1 were highly expressed. Conversely, when the first 6 days were in darkness, specific IAA oxidase activity was high and the isoforms pI 4.5, 6.7 and 6.8. were most strongly expressed. The isoperoxidases pI 7.8 and 7.9 were strongly expressed when the explants were cultured for at least 6 days in darkness.     

Changes of Endogenous Hormone Contents During Floral Bud and Vegetative Bud Differentiation in Thin Cell Layer Culture of Cichorium intybus L. Explant

The sectioned thin cell layers (TCL) of flower stalk of Cichorium intybus L. were cultured in MS medium supplemented with NAA and BA or IAA and BA where floral and vegetative buds were developed from the explant. Endogenous IAA, DHZ+DHZR, iPA increased significantly during the floral bud formation, while Z+ZR remained changed. The levels of cytokinins, DHZ +DHZR, iPA, and Z-f-ZR all increased significantly during the vegetative bud formation, however IAA level was reduced during the first 7 days of culture and increased to two-thirds of initial values on the day when the bud primordia were formed. The results suggested that the initiation of floral buds was associated with a high IAA/CTK ratio, whereas the induction of vegetative bud differentiation was related to a low IAA/CTK ratio.     

菊苣薄层培养花芽,营养芽分化中内源激素的动态变化

菊苣（Ｃｉｃｈｏｒｉｕｍ ｉｎｔｙｂｕｓＬ．）花梗薄层细胞培养于ＭＳ附加ＮＡＡ 和ＢＡ 或ＩＡＡ 和ＢＡ 的ＭＳ培养基上有花芽或营养芽分化． 花芽分化中内源ＩＡＡ、ＤＨＺ＋ ＤＨＺＲ、ｉＰＡ 含量明显增加,而Ｚ＋ ＺＲ变化不明显．营养芽分化中内源细胞分裂素含量增加明显,而ＩＡＡ 在培养前７ ｄ 含量下降,随后有所增加,在原基形成时含量达原初水平的２／３． 可见,花芽分化比营养芽分化所需内源ＩＡＡ／ＣＴＫ 比值要高     

In vitro propagation of Cichorium intybus L. and quantification of enhanced secondary metabolite (esculin)

In this report, rapid and effective shoot as well as root regeneration system through direct multiplication was successfully developed for Cichorium intybus L. Furthermore, the effect of exogenous growth regulators (TDZ and IAA) at different concentrations on the regulation process of the plant was also studied. Enhanced production of esculin in developed C. intybus L. was evaluated using leaf extract. Only on the expense of 20 days, regeneration was seen and very low dose of TDZ was seen to be more effective. When 0.02 mg/L of TDZ was combined with 1.5mg/L of IAA, nearly 100% of explants produced shoots with the highest number of regenerated shoots (85.37). With further increase in concentration (≥ 0.05 mg/L), the number of shoots per explants get decreased. A lower NAA to IBA ratio (1.0mg/L of IBA and 0.5mg/L of NAA) seemed to be more effective for root generation and considered to be the most effective combination among the tried groups. IBA was more effective in root development than NAA, but both were comparatively effective. On quantitative analysis by RP-HPLC, the 76.23% of Esculin were found in leaf extract of the in vitro developed C. intybus L. This amount was 26.77% higher than normal grown plants.     

Endometrial protein secretion during early pregnancy in entire and ovariectomized ewes

The secretion and synthesis of protein in vitro by explants of endometrium were examined in entire ewes during the first 10 days of the oestrous cycle and during an equivalent interval in ovariectomized ewes which received injections of oestradiol and progesterone. The schedule of steroid injections given was designed to simulate endogenous ovarian secretion of progesterone during the luteal phase before oestrus, of oestradiol around oestrus and of progesterone during the luteal phase after oestrus. The rate of protein synthesis and tissue RNA:DNA and protein:DNA ratios in intercaruncular and caruncular endometrium were generally higher in entire than in ovariectomized ewes. In ovariectomized ewes oestradiol increased these activities at 2-4 days after oestrus, whereas progesterone preceding oestradiol caused increases at oestrus, but not thereafter. In entire ewes and in ovariectomized ewes receiving the full steroid treatment regimen, protein secretion was high at oestrus and declined markedly during the next 4-6 days. In ovariectomized ewes not receiving progesterone before oestradiol, secretion increased between 4 and 6 days after oestrus, or during the equivalent stage of treatment in ewes which did not show oestrus. The omission of this progesterone did not modify secretion by caruncular endometrium. Oestradiol increased protein secretion by both tissues. The data suggest that progesterone given before oestradiol (or its equivalent in entire ewes) inhibits the secretion, at about 4-7 days after oestrus, of uterine proteins which may impair embryo development in ovariectomized ewes which do not receive this progesterone.     

紫花苜蓿和菊苣比叶面积和光合特性对不同用量保水剂的响应

为研究紫花苜蓿(Medicago sativa)和菊苣(Cichorium intybus)叶片生长和光合生理对不同用量保水剂的适应能力和生理响应机制,以不施保水剂作为对照(CK),测定了施用不同用量保水剂(15 kg · hm~(-2),30 kg · hm~(-2))条件下大田种植的紫花苜蓿(Medicago sativa)和菊苣(Cichorium intybus)的比叶面积和光合特性.结果表明,保水剂对紫花苜蓿的比叶面积无显著影响(P>0.05),但对菊苣的比叶面积具有显著影响(P<0.05).对紫花苜蓿而言,保水剂施用量为15 kg · hm~(-2)时,其P_n、g_s和WUE均显著高于对照(P<0.01),但施用量为30 kg · hm~(-2)处理时,其P_n、和g_s均显著低于对照处理(P<0.01),但是却维持较高的WUE;对菊苣而言,施用保水剂降低了其P_n和T_r(P<0.01),但WUE与对照无显著差异(P>0.05),这表明紫花苜蓿和菊苣采取两种不同的生理适应策略来适应变化了的水分环境条件.紫花苜蓿P_n与g_s、P_n与T_r的相关性均以保水剂施用量为30 kg · hm~(-2)处理最紧密(P<0.01),而P_n与WUE的相关性则以对照最为紧密,依次为15 kg · hm~(-2)处理和30 kg · hm~(-2)处理;菊苣P_n与g_s、Tr和WUE的相关性强弱均表现为15 kg · hm~(-2)处理<30 kg · hm~(-2)处理<对照,表明保水剂影响叶片气体交换参数之间的关系.研究表明,不同植物叶片参数和光合特性对保水剂的响应并不具有一致性,植物可以根据环境条件的变化调节其形态和生理过程,以维持其正常生长.     

巨桉凋落叶分解对菊苣生长及光合特性的影响

采用盆栽试验,研究了巨桉凋落叶分解初期对菊苣幼苗生长和光合生理特性的影响.试验设置A₁(30 g·pot^-1)、A₂(60 g·pot^-1)、A₃(90 g·pot^-1)和对照（CK）4个凋落叶水平,将各处理的凋落叶分别与12 kg土壤混合后装盆,播种菊苣.待A₃处理植株的第3片真叶完全展开后测定菊苣光合生理指标及相关生长指标.结果表明：巨桉凋落叶分解初期明显抑制了菊苣生物量积累、叶面积的增加及光合色素的合成,且随着凋落叶施入量的增加抑制作用加大;菊苣叶片胞间CO₂浓度增加,而净光合速率、气孔导度和蒸腾速率均显著低于对照;随土壤中凋落叶含量的增加,除CO₂补偿点呈增加趋势外,其他光响应和CO₂响应的特征参数都呈明显下降趋势,并与对照差异显著.巨桉凋落叶分解过程中,其化感物质逐步释放并作用于受体植物,抑制其光合色素合成和光合作用,降低其环境适应能力,从而抑制菊苣的生长.     

The Changes of Protein Content and Synthetic Activity in Squash Pollen During Germination

The total protein content of squash (Cucurbita moschata Duch.) pollen decreased gradually during in vitro germination. It was caused by the release of wall proteins and part of the cytoplasmic proteins. The release of the pollen wall proteins was not dependent on germination, it was a passive diffusion process. However, the cytoplasmic proteins did not release until the pollen germinated, a fraction of them was synthesized de novo during germination. The RNA and protein synthetic activities initiated soon after in vitro pollen germination. The RNA synthesis decreased during germination. As about half the activity was inhibited by α-amanitin, mRNA might be the major RNA synthesized de novo. The total protein synthesis increased during germination, almost all of this synthesis was inhibited by cycloheximide, and partially by α-amanitin, but it was not affected significantly by actinomycin D. These results indicated that both stored and de novo synthesized mRNA might play a role in the protein synthesis. The content of stored mRNA of squash pollen was about 11-3 pg/grain as measured by UV absorption after its purification from total RNA (2440 pg/grain) by oligo (dT)-cellulose affinity chromatagraphy. Both cycloheximide and α-amanitin inhibited pollen tube growth in vitro. Actinomycin D and tunicamycin inhibited pollen germination in the first hour, however, no reduction ,of the tube length was observed later. Cyclohex,nide inhibited the pollen germination and tube elongation in vivo, that fitted well with the in vitro results. According to these results, it was suggested that the de novo syntheses of mRNA and protein were neccessary for the maintenance of pollen tube growth.     

Nucleic acid synthesis accompanying the recovery of cell division and chloroplast development in "giant" cells of the Emerson strain of Chlorella

The light-induced recovery of cell division and chloroplastdevelopment in "giant", "bleached", cells of the Emerson strainof Chlorella takes place without any increase in DNA and isrelatively insensitive to mitomycin C and 5-bromouracil. 5-Fluorouracilinhibits cell division only when it is supplied during the earlystages of recovery, perhaps by interfering with that phase ofRNA synthesis which occurs during the first 6 hr of recovery.This early burst of RNA synthesis is more sensitive to chloramphenicolthan is the second phase of RNA synthesis, suggesting that asignificant proportion of it may originate in the chloroplast.Evidence is presented which suggests that 5-fluorouracil interfereswith chloroplast development primarily through an effect onchlorophyll synthesis. The possible significance of these observationsin relation to nuclearchloroplastic interractions is discussed. (Received December 15, 1972; )     

Guaianolides from Cichorium intybus and structure revision of Cichorium sesquiterpene lactones

The isolation and structure elucidation of a new lactucopicrin derivative from Cichorium intybus is described, together with the revised structures of several sesquiterpene lactones previously isolated from Cichorium species. The known eudesmanolide magnolialide and the known guaianolide ixerisoside D, reported for the first time from the plant material, along with the previously known chicory sesquiterpene lactones have been also isolated and identified.