Control of the activity of plant plasma membrane MgATPase by the viscosity of the aqueous phase

The activity of plant plasma membrane (PM) MgATPase (EC 3. 6. 1. 35) was studied in PM vesicles purified from spring wheat (Triticum aestivum L. cv. Drabant) roots, winter wheat (Triticum aestivum L. cv. Martonvásári-8) roots, and soybean (Glycine max L. cv. Williams) hypocotyls by aqueous polymer two-phase partitioning. MgATPase from spring wheat roots was assayed at 23°C (a) in the absence and presence of Triton X-100, (b) in the presence of either 1 mM or 3 mM MgATP^2?, and in the presence of increasing concentration of sucrose from 10 mM upto 1. 2 M. Activity of MgATPase in PM vesicles from winter wheat roots was measured at 21°C at sucrose concentrations of 15 mM–1. 1 M in the presence of 3 mM MgATP^2? and absence of Triton X-100 only. MgATPase activity from soybean hypocotyls was studied (a) in the absence and presence of Triton X-100, (b) both at 21 and 31°C, in the presence of 3 mM MgATP^2? and varying concentrations of sucrose between 10 mM and 1. 1 M. In all cases, independently of the assay conditions and the source of PM, the MgATPase activity decreased with increasing sucrose concentration. Latency of the MgATPase activity depended only slightly on the concentration of sucrose. The Q₁₀ value for the MgATPase activity from soybean hypocotyls (and thereby the Arrhenius activation energy of the enzymatic reaction) was independent of the sucrose concentration and of the presence of Triton X-100. At optimal assay conditions, the ATP-hydrolyzing activity of plant PM MgATPase (v) was inversely proportional to the m-th power of the viscosity of aqueous phase (η) as predicted by the modified Kramers'theory of enzymatic catalysis: v ∝ (1/η)^m, where m is an empirical parameter between 0 and 1. For the activity of MgATPase in the three species studied, m varied between 0. 5 and 1. 1 in good agreement with the theory. We thus conclude that (a) the activity of integral membrane enzyme-proteins may be controlled not only by the property(ies) of the membrane lipid phase but also by the viscosity of the aqueous phase in the vicinity of such enzymes, and (b) the determination of vesicle sidedness based on enzyme latency may need a minor revision. Our interpretation is in agreement with the molecular dynamics approach of enzymatic catalysis worked out for soluble enzymes.     

蔗糖诱导拟南芥幼苗胚轴维管束细胞数量增多

为揭示蔗糖能否引起植物胚轴维管束细胞数量增多，将拟南芥播种于添加88mmol·L-1蔗糖和不添加糖的MS培养基上，对生长在不同培养基上的幼苗胚轴横切，显微镜下统计切片上维管束细胞数量。结果显示，与不加糖相比，加糖条件下萌发4d后幼苗维管束细胞总数增加约70％，维管薄壁细胞和导管分子都增加100％以上，筛管分子增加约90％，中柱鞘细胞数量不变。显然，蔗糖不仅使维管束薄壁细胞数量增多，也使筛管分子和导管分子数量增多。因此认为，添加蔗糖对拟南芥幼苗胚轴维管束具有双重效应，既引起维管薄壁细胞增殖，又促进维管薄壁细胞分化，从而使导管分子和筛管分子数量增多。     

Direct shoot organogenesis and plant regeneration in Fortunella crassifolia

An efficient in vitro regeneration system in kumquats (Fortunella crassifolia Swingle) was established. Explant types and orientations, concentrations and combinations of plant growth regulators were evaluated for their influences on efficiency of plant regeneration. It was found that the optimum explant and its orientation was epicotyl planted vertically with upper part upward, and a shoot regeneration frequency of 1.48 shoots per explant was obtained on Murashige and Skoog (1962; MS) medium supplemented with 22.19 μM 6-benzyladenine (BA). A rooting percentage as high as 74 % was obtained on 1/2 MS supplemented with 0.54 μM 1-naphthaleneacetic acid (NAA), 9.29 μM kinetin (KN), and 0.5 g dm⁻³ activated charcoal.     

The Connection between High Energy and Low Energy Responses to Red and Far-Red Light of Hypocotyl Elongation in Red Cabbage Seedlings (Brassica oleracea var. capitata'Marner Septemberrot')

Growth responses of hypocotyls of Brassica oleracea L. var. capitata caused by diurnal irradiation of various duration (0.5 h/d up to continuous irradiation) and various irradiance (0.3–1.5 W m^?2) with red and far-red light from sources characterized below can be described by the equation: (W= relative effect, I= irradiance, t₁= irradiation time per day, W_s= constant dependent on material and wavelength, α, B and t_s, = constants dependent on wavelength.) On the basis of that equation a possible connection between longtime and short-time irradiation responses is discussed.     

A rapid protocol for in vitro micropropagation of Lepidium sativum linn. and enhancement in the yield of lepidine

Summary Lepidium sativum L., commonly known as ‘garden cress’ possesses variable proportions of benzylcyanide and benzylisothiocynate which contribute towards its activity against Bacillus subtilis and Micrococcus pyogenes. The plant is also used as an antifertility and antiovulatory drug. Various juvenile (cotyledonary leaves, hypocotyl, radicle) as well as mature explants (leaf, shoot apex, nodal segments) callused on Murashige and Skoog's medium (MS) supplemented with α-naphthaleneacetic acid (NAA)+N⁶-benzyladenine (BA)+casein hydrolyzate (CH). Regeneration from hypocotyl callus and nodal segments occurred after NAA/BA was replaced with indole-3-acetic acid (IAA)/kinetin (Kn). Lepidine was monitored at regular intervals. Significant amounts of lepidine was detected in in vitro-regenerated plants obtained from juvenile and mature explants. The yield, however, was variable, depending upon the source and type of explant used. High lepidine was detected in 8-wk-old hypocotyl callus. Amongst regenerants, maximum lepidine was obtained from the plantlets at the vegetative stage.     

Plant cell remodeling by autophagy

Plant seedlings are not photoautotrophs until they are equipped with photosynthetic machinery. Some plant cells are remodeled after being exposed to light, and a group of peroxisomal proteins are degraded during the remodeling. Autophagy was proposed as one of the mechanisms for the degradation of peroxisomal proteins. We recently showed that ATG7-dependent autophagy is partially responsible for the degradation of obsolete peroxisomal proteins during Arabidopsis seedling growth.     

Phototropism in Higher Plants — Controversies and Caveats1

Recent literature on light-induced changes in the distribution of growth and of endogenous growth regulators in the development of phototropic curvature is reviewed. It is concluded that in a great many cases the Cholodny-Went hypothesis is sufficient to account for the growth changes bringing about curvature, though in certain dicotyledonous seedlings differential effects of light on the synthesis of growth inhibitors across the tissue from the lighted to the shaded side are implicated. The problems in the interpretation of results from experiments in which more than one photoresponse may be simultaneously induced are discussed and methods of circumventing them considered. Action spectroscopy suggests that dicotyledonous seedlings may have the same phototropic photoreceptor as monocotyledonous seedlings.     

Control of in vivo protein synthesis and peroxidase formation by DNA-containing extracts of normal and crown gall tissues

Nucleic acids extracted from normal bean hypocotyl tissue (NE) and crown gall tumors (TE) affect amino acid incorporation into protein and the development of peroxidase activity when vacuum infiltrated into normal receptor tissues. TE enhances and NE inhibits both processes; NE from successively older tissues produces progressively greater inhibitions per unit of infiltrated nucleic acid. The active material has an absorption maximum at 257 nm with an A_260:280 ratio of more than 2·0. On acrylamide gel electrophoresis it shows a small DNA peak, four typical r-RNA peaks and a small low molecular weight RNA peak. Activity in such extracts is completely destroyed by hydrolysis with 0·3 N KOH or DNAase; RNAase is only slightly effective and pronase ineffective. It is deduced that the effective material contains DNA that may be complexed with RNA or other materials in the extract. Pretreatment of donor tissues with actinomycin d or 5-fluorouracil diminishes or annuls the activity of the extract. Pretreatment of receptor tissue with actinomycin d inhibits the action of TE but not of NE; pretreatment with cycloheximide prevents the action of both NE and TE.     

Micropropagation of juvenile and adult Annona squamosa

A micropropagation system for Annona squamosa L. (Sugar Apple) using hypocotyls of seedlings and nodal cuttings from 3-year-old plants was developed. Shoot proliferation was achieved with Woody Plant Medium supplemented with BA. Silver thiosulphate was added at 0.5 mg l^–1 to control leaf abscission. Rooting was obtained when subcultured shoots were preconditioned for 2 weeks in medium with 10 g l^–1 activated charcoal before treatment with 43 µm NAA or 39 µm IBA. Rooting was improved when galactose was used instead of sucrose in the rooting medium. The rooted plantlets were acclimatised successfully.Abbreviations NAA naphthaleneacetic acid - IBA indolebutyric acid - MS Murashige & Skoog Medium - WPM Woody Plant Medium - NN Nitsch Medium - Juv juvenile explant - Adu adult explant