A Study of the Growth of Excised Embryo Shoot Apices of Wheat In vitro

A technique for obtaining excised embryo shoot apices of wheatis described. When yeast extract or various amino acids wereincluded in the basal medium the explants did not grow and muchof the tissue showed signs of necrosis. Coconut milk, eitherautoclaved or filter-sterilized, inhibited the growth of theexplants. Root initiation and shoot growth were stimulated byammonium salts. It was not known whether root initiation wasthe result of stimulation by ammonium salts or the rapid growthof the shoot apex. Aqueous extracts of roots also stimulatedthe growth of the explants, but extracts of other plant organsdid not. The addition of individual vitamins had various effectson the growth of the explants, but since the growth responsewas so slow, results other than complete inhibition were rejected.On the basis of these results, other substances appear to benecessary to support the growth of wheat explants without theinitiation of roots.     

Growth in Axenic Culture of Isolated Shoot Apices of Eichhornia crassipes

Apical meristem culture of Eichhornia crassipes has shown that for successful regeneration, the excised meristem dome must be associated with at least the youngest leaf primordium as part of the explant and a culture medium containing coconut milk (10 %, v/v), IAA (0.1 mg/l) and kinetin (1 mg/l) as growth supplements with 2 % sucrose as carbon source.     

Culture of Shoot Apices of Theobroma cacao

Surface sterilized buds of young cocoa plants (Theobroma cacao L.) taken at particular stages of the flush cycle were placed in Linsmaier and Skoog agar medium supplemented with a range of growth regulators. Only buds taken at the 1–2 (dormant) stage of the flush cycle and treated with gibberellic acid (GA₃) alone and GA₃plus kinetin (KN) supplement showed bud opening. In liquid Linsmaier and Skoog medium buds isolated at the 1–2 stage also responded to GA₃ and KN. In this case addition of KN caused bud opening, while GA₃ either initiated bud opening only or opening followed by leaf expansion depending on the concentration of GA₃ supplied. Bud development was inhibited when ABA was included in the medium hut this was overcome by the presence of GA₃ but not KN. Since a hormonal supplement was required for any response from the excised buds, it is suggested that the intermittent growth of the shoot apex in the intact plant may be determined by hormonal stimuli derived from other parts of the plant. The findings also indicate that the bud apices could be maintained in culture for long periods which may provide a basis for the development of a micropropagation procedure for cocoa.     

The Effect of Growth Substances on Growth of Excised Embryo Shoot Apices of Wheat in Vitro

Excised embryo shoot apices of wheat were grown on media containingeither indolylacetic acid (IAA), kinetin, gibberellic acid (GA),or adenine. GA and adenine stimulate the activity of the apicalmeristem whereas IAA and kinetin do not. No combination of anytwo substances significantly stimulated growth when comparedwith the most active substance present. GA can overcome theinhibitory effect of IAA or kinetin. The possible function ofthese substances in the apical meristem is discussed.     

Connective Auxin Transport in the Shoot Facilitates Communication between Shoot Apices

The bulk polar movement of the plant signaling molecule auxin through the stem is a long-recognized but poorly understood phenomenon. Here we show that the highly polar, high conductance polar auxin transport stream (PATS) is only part of a multimodal auxin transport network in the stem. The dynamics of auxin movement through stems are inconsistent with a single polar transport regime and instead suggest widespread low conductance, less polar auxin transport in the stem, which we term connective auxin transport (CAT). The bidirectional movement of auxin between the PATS and the surrounding tissues, mediated by CAT, can explain the complex auxin transport kinetics we observe. We show that the auxin efflux carriers PIN3, PIN4, and PIN7 are major contributors to this auxin transport connectivity and that their activity is important for communication between shoot apices in the regulation of shoot branching. We propose that the PATS provides a long-range, consolidated stream of information throughout the plant, while CAT acts locally, allowing tissues to modulate and be modulated by information in the PATS.     

玉米芽尖培养中的高频率体细胞胚胎发生与植株再生(简报)

通过诱导玉米芽尖产生胚性愈伤组织,建立起高频率植株再生的玉米芽尖培养实验体系。在MS+1.0mg·L-16-BA+0.2mg·L-12,4-D+500mg·L-1CH的培养基上诱导愈伤组织并继代培养1次后,将愈伤组织转移到Ms+0.5mg·L-16-BA+0.4mg·L-1IBA+500mg·L-1CH的分化培养基上,可形成大量的体细胞胚胎。组织学观察表明,体细胞胚胎主要发生在胚性愈伤组织表面与表层下部。不同基因型的芽尖形成愈伤组织和再生植株的能力不同。     

The Growth of Shoot Apices and Inflorescences of Carex flacca Schreb. in Aseptic Culture

The vegetative apex and young inflorescence of Carex flaccahave been grown in aseptic culture for several weeks on a definedmedium. Explants comprised the apical dome and the three youngestleaf primordia, and the young inflorescence complete with bracts,excised shortly after the initiation of the lateral spikes.Some growth occurred on the basal medium which contained inorganicsalts, sucrose, and vitamins, but growth was increased and thelife span extended by the addition of certain other compounds.The most effective additive was gibberellic acid, which, however,resulted in precocious differentiation of the meristematic tissuesand the differentiation of abnormal xylem. These deleteriouseffects of gibberellic acid were counteracted by the additionof kinetin. This substance did not otherwise affect vegetativeapices but it resulted in a further increase in growth and lifespan of inflorescences.     

Characterization of Stelar Initiation in Shoot Apices of Ferns

Procambium is commonly recognized as a vascular meristem inshoot apices of vascular plants. Prestelar tissue comprisingprovascular tissue (PVT) and pith mother cells (PMCs) immediatelysubjacent to the single cell layer of promeristem has been consideredto represent the initial stage of stelar differentiation precedingprocambium and rib meristem in ferns. In addition to characterizationof PVT and PMCs on the basis of cell morphology, cytologicalfeatures and developmental continuity with procambium and ribmeristem, four lines of evidence from studies of shoot apicesof Matteuccia struthiopteris and Osmunda cinnamomea supportthis interpretation of initial differentiation. (1) Differentialstaining by safranin-fast green and crystal violet-erythrosinshows that PVT and PMCs differ in colour reactions from promeristemand resemble procambium and pith meristem, respectively. (2)Comparative ultrastructural study reveals qualitative differencesin the cell membrane system, nuclei, cytoplasm, vacuoles andplastids between promeristem and PVT but similarity of PVT toprocambium. (3) Large droplets of tannins occur in promeristembut not in PVT, PMCs and procambium. (4) Cytochemical studyof the shoot apex of Osmunda shows that carboxylesterase activityis strongly demonstrated in PVT and procambial cells but notin promeristem cells and PMCs. These observations further substantiatethe interpretation that PVT represents initial vascular differentiationand PMCs reflect a commitment to pith development.Copyright1995, 1999 Academic Press Initial vascular differentiation, provascular tissue, differential staining, ultrastructure, tannins, carboxylesterase, shoot apex, Matteuccia struthiopteris, Osmunda cinnamomea     

Organogenesis in Callus from Shoot Apices of Pisum sativum

Shoot formation was observed in callus from apical cells of pea (Pisum sativum L. cv. Century). Shoot apices from 4-day-old plants were macerated and the resulting cell masses grown on agar media. The callus formation and shoot production occurred within 4 to 6 weeks in defined media containing 0.2 to 5.0 μM benzyladenine and 1 μM naphthaleneacetic acid. While most callus produced one or more shoots at high frequency, root formation did not occur regularly. Plants obtained by these procedures were grown to maturity producing flowers and pods.     

Ascorbate Oxidase Activity in Rice Shoot Apices during Panicle Initiation

There was a decrease in the ascorbate oxidase activity in theshoot apices of Oryza sativa L. cv BAM 11, a photo-sensitiverice variety. Ascorbic acid content in the shoot apex increasedand exogenous application of ascorbic acid resulted in an earlinessin panicle initiation. Photoperiodic treatment promoted panicleinitiation with an early and faster decline in ascorbate oxidaseactivity. Decline in the activity of ascorbate oxidase showeda negative correlation with the increase in ascorbic acid content. (Received November 11, 1985; Accepted February 17, 1986)     

Cleared Whole Mounts of Shoot Apices of Angiosperms for Topographic Study

Cleared and stained whole mounts of stem apices of two Labiates and of Phaseolus plumule giving a three-dimensional picture of the apical structure have been prepared as follows. Fix the buds in formalin-acetic acid-50% alcohol (5:5:90) for 24 hr or longer and then dissect under a binocular microscope to leave only the youngest leaves surrounding the apex. Wash for several minutes in distilled water and then clear the material in a 5% solution of sodium hydroxide at approximately 40° C for 24-48 hr. Wash thoroughly in several changes of distilled water, transfer to a solution of 1% tannic acid and 0.5% sodium salicylate for up to a minute. Wash briefly in distilled water and stain in a 1.5% solution of ferric chloride until blue-black. Wash in distilled water and dehydrate through 50%, 70%, 85%, 95% and 2 changes of absolute ethyl alcohol. If the xylem is not stained well, counter-stain for a few seconds in a 0.5% solution of safranin O in a 1:1 mixture of xylene and absolute alcohol and wash out the excess stain in the same mixture. Clear in 2 changes of xylene and place on a glass slide in thick Canada balsam. Orient with needles under low magnification and cover.     

In vitro Growth and Shoot Multiplication of Achras zapota in a Controlled Carbon Dioxide Environment

The culture vessels with multiplying shoots of Achras zapota L. on Schenk and Hildebrandt (SH) medium containing 8.88 M 6-benzylaminopurine (BAP) with or without sucrose were kept under varied CO₂ concentrations ranging from 0.6 to 40.0 g m^–3 using different concentrations of sodium bicarbonate (NaHCO₃), sodium carbonate (Na₂CO₃), potassium bicarbonate (KHCO₃), and potassium carbonate (K₂CO₃) in small acrylic chambers. Complete absence of carbon source caused death of shoots within 20 d. Under elevated concentrations of CO₂ (10.0 and 40.0 g m^–3) the shoots grew photoautotrophically on sucrose-free medium. The growth of cultures was better at 40.0 g (CO₂) m^–3 than on 3.0 % sucrose under ambient air of growth room. However, the best response was obtained at 10.0 g (CO₂) m^–3 and 3.0 % sucrose where maximum number of shoots, shoot length, fresh and dry mass, total number of leaves and leaf area was observed.     

Development During Vegetative Growth in the Apical Region of the Shoot of Lupinus albus

This investigation was designed to show the nature of the developmentin terms of cell numbers and of rates of division that proceedsin the terminal region of the shoot during the vegetative phasein Lupinus albus. Data are assembled that show that with eachsuccessive plastochron in the dome the number of cells increasesand the rate of division decreases; in the first internode againthe number of cells increases and the rate of division decreases,and in the first primordium both the number of cells and therate of division decrease. The data also show that with thetransition from one plastochron to the next the rate of divisionin any particular intemnode or primordium decreases. It is asignificant feature that all the changes in the system can becharacterized through constant numerical factors. The significanceof the results is discussed.     

Germplasm Preservation of Wild Arachis Species through Culture of Shoot Apices and Axillary Buds from In Vitro Plants

A study was conducted to evaluate in vitro techniques for germplasm preservation of wild species of Arachis. Nodal segments excised from in vitro-grown plants of A. retusa, A. macedoi and A. burchellii were used to examine the effects of explant position and age of the donor plant. Explants were excised from plants maintained in culture for 30, 60, 90 or 180 d, numbered I – V from top to bottom and cultured on MS medium supplemented with 2.7 µM NAA or different BAP concentrations (0, 4.4, 13.2 and 22 µM). The age of the donor plant has not influenced the responses of the four genotypes studied. In contrast, shoot regeneration ability was significantly affected by the original explant position, decreasing from top to bottom. In media supplemented with different BAP concentrations, multishoot formation was induced from apical segments at low frequencies (10 – 20%) and segments of all positions originated calluses at the explant basis after 30 d of culture. The culture of nodal segments in the presence of 2.7 µM NAA as the sole growth regulator is recommended for the multiplication of in vitro collections of wild groundnut species in order to avoid callusing and adventitious shoot formation.     

Staining Angiosperm Shoot Apices with Heidenhain's Iron Hematoxylin and Aniline Blue

Good differential staining of nuclei, cytoplasm and cell walls of angiosperm shoot apices is obtained by a hematoxylin-aniline blue sequence. First, follow a typical Heidenhain's iron-hematoxylin scheme so that the nuclei and cytoplasm are well stained, then bring the slides up through an ethyl alcohol series to absolute alcohol. Transfer to a saturated solution of aniline blue in methyl cellosolve for 10 minutes. Remove the excess aniline blue with absolute alcohol, and follow this with a mixture of 42 parts absolute alcohol, 25 parts methyl salicylate, and 33 parts xylene; next, a similar mixture but in the proportions of 1:2:1; then a xylene-alcohol mixture, 9:1, followed by pure xylene, 2 changes, and covering in balsam. Panchromatic plates or film are suitable for photomicrographic reproduction (Ilford Special Rapid Panchromatic plates were used).     

Surface Changes on Fresh Carnation Shoot Apices During Scanning Electron Microscopy

The general morphology of fresh intact shoot apices of carnation,Dianthus caryophyllus L. cv. Improved White Sim was studiedwith a scanning microscope. The changes in the surface withtime under the electron beam were: (1) individual cells beingvisible, (2) a smooth apex and (3) wrinkling until the specimenbecame unsatisfactory for study. As indicated by time coursestudies, the initial individual cell observations representthe true features of the intact apex. The observation of a smoothapex was related to a ballooning of the surface layer resultingfrom water vaporization. Incisions in the apex to prevent pressurebuild-up under the surface layer revealed the smooth characteristicof the apex surface was due to lifting of the cuticle from underlyingcells. Areas adjacent to a micropuncture in the apical domedid not exhibit ballooning.