Studies on the plant cytoskeleton using miniprotoplasts of tobacco BY-2 cells

Vacuoles in plant cells can be eliminated by centrifugation of protoplasts through a density gradient. In this review, properties of evacuolated protoplasts, named ‘miniprotoplasts’, and the significant roles in plant cytoskeleton studies are described. Miniprotoplasts, prepared from tobacco BY-2 cells whose cell-cycle had been synchronized at late anaphase, continued to divide to form two daughter cells. In the presence of cytochalasin B cytokinetic cleavage was enhanced, suggesting a role of actin filaments in plant cytokinesis. In the cytoplasmic extract of miniprotoplasts both tubulin and actin could be polymerized to form microtubules (MTs) and actin filaments (AFs), respectively. A purification method for tubulin, actin and related proteins was developed using the extract. To investigate the interaction between cortical microtubules and the plasma membrane, an experimental system in which MTs were reconstructed on membrane ghosts was developed by combination of membrane ghosts and the extract.     

Purification and characterization of ferredoxin-sulfite reductases from leek (Allium tuberosum) leaves

Ferredoxin-sulfite reductases (Fd-SiRs) [hydrogen-sulfide: ferredoxin oxidoreductase, EC 1.8.7.1] from leek leaves have been purified to homogeneity. The enzymes (SiR 1, SiR 2 and SiR 3) were separated by Mono Q chromatography. The collective molecular mass of the enzymes was estimated to be 65 kDa by gel filtration. In all three cases, subunit analysis by SDS-PAGE yielded a single protein band corresponding to a molecular mass of 64 kDa, indicating that the enzymes each exist as a monomer. In the oxidized forms, SiR 1, SiR 2 and SiR 3 all exhibited nearly identical absorption maxima at 279∼280, 389∼390, 588 and 714 nm, indicating that siroheme is involved in the catalysis of sulfite reduction. On enzymatic properties, SiR 1, SiR 2 and SiR 3 could only react with the physiological electron donor, feriedoxin. The enzymes exhibited different heat stabilities. The pH active curve obtained from SiR 2 was different from the others. Moreover, SiR 1 exhibited a lower Km value for ferredoxin than SiR 2. Although the N-terminal sequence was the same, the results of some enzymatic properties, amino acid analysis, and peptide mapping suggested the presence of the Fd-SiR isozymes in leek leaves.     

The vegetation of the Puna Belt at laguna de Pozuelos Biosphere Reserve in northwest Argentina

The presence of species and their cover values together with soil and topographic characteristics were recorded at 40 sites along an altitudinal gradient of the Puna Belt at Pozuelos, in the High Andes in northwest Argentina. Classification and ordination of plant assemblages showed that soil and topography were the best predictors of the variation in species distribution. The different plant assemblages which occur along the topographic gradient follow the geographic variation in rainfall in this Andean region. Perennial bushes and grasses colonize the plains and slopes with fine textured soils which retain their water content. In contrast, deciduous bushes and cactuses grow on steep, rocky, dry slopes. Open woods also occur on east-facing steep, rocky slopes which are sheltered from winds.     

长白山劲松林场植物群落的分类和排序

将长白山白河林业局劲松林场的５４块样地用聚类分析法划分成６个植被类群,再用ＰＣ－ＶＴＡＢ程序中经过改进的Ｂｒａｕｎ－Ｂｌａｎｑｕｅｔ学派的植被排表分析法进行综合,产生了鉴别概要表,为各个等级的植被类群筛选出诊断种。此外,还用鉴别种地样地记录进行主成分分析,以验证诊断种的有效性,结果表明,ＰＣ－ＶＴＡＢ中的植被排表分析法是筛选鉴别种的有效方法,而鉴别种以显著地提高植被分析和排序的质量。     

可乐茄叶肉原生质体培养再生植株

本文报道茄属果树可乐茄（ＳｏｌａｎｕｍｑｕｉｔｏｅｎｓｅＬａｍ．）叶肉原生质体的分离、培养及植株再生。幼嫩叶片原生质体经酶游离、纯化后,以１×１０４个／ｍｌ密度培养于稍加改良Ｋ８ｐ（附加２,４－Ｄ０．５ｍｇＬ（－１）、ＮＡＡ１．０ｍｇＬ（－１）和ＢＡ０．５ｍｇＬ（－１））的培养基中,三天后开始分裂,一周分裂３—４次。一个月形成小细胞团,植板率为０．１—０．２％,小细胞团转培养于ＭＳ＋２,４－Ｄ０．５ｍｇＬ（－１）上增殖后进行分化。原生质体来源愈伤组织在ＩＡＡ（０．１—１．０ｍｇＬ（－１））与ＢＡ或ＺＴ组合的培养基中能诱导器官发生,芽分化率最高可达４２．９％;但ＩＡＡ、ＢＡ、ＺＴ三者一起使用未见任何器官分化。小芽在ＭＳ＋ＩＡＡ０．２ｍｇＬ（－１）中生根成植株。可乐茄叶肉原生质体的植株再生,可应用于育种和茄属植物遗传工程研究。     

The influence of habitat structure on arthropod diversity in Argentine semi-arid Chaco forest

Abstract. The Argentine Chaco is a mosaic of grassland and open forest habitats maintained by natural disturbance activities such as fire. Since the introduction of domestic livestock and other human activities, the balance of this mosaic has been significantly altered, both in plant species and structural composition. This study focuses on the impact of such changes on the diversity of ground-dwelling arthropods within semi-arid Chaco forest. Quantitative measures of habitat structure and arthropod diversity were taken in forest areas previously subjected to grazing, logging and ploughing. Results indicated that arthropod diversity was smaller on sites with reduced structural complexity, with marked changes in arthropod family composition. The habitat components relating to plant architectural and vertical diversity were particularly influential on arthropod diversity. The guild size ratio of predatory to non-predatory arthropods also differed significantly between habitats suggesting a change in the resource base available to some groups. The latter suggests a shift in the functional organisation of the forest ecosystem which could have important repercussions for the diversity of other trophic levels.     

Fertile plant regeneration from protoplasts of a seed-propagated cultivar of Lilium × formolongi by utilizing meristematic nodular cell clumps

Plant regeneration from protoplasts of Lilium × formolongi cv. Azusa was achieved by utilizing suspension cultures of meristematic nodular cell clumps with a high plant regeneration ability. Creamy-white calli with embryogenic potential were initially induced from the seeds on Murashige and Skoog (MS) medium containing 4.1 μM picloram. The calli were then transferred into a liquid medium of the same composition, in which they turned into compact cell clumps which consisted of meristematic nodules. Protoplasts were readily isolated from these meristematic nodular cell clumps. Colonies were successfully formed from the protoplasts by embedding in 0.1% gellan gum-solidified MS medium containing 4.1 μM picloram and 0.5 M glucose. They regenerated shoots and roots on MS medium containing 2.2 μM benzylaminopurine (BAP). The plants thus obtained produced flowers with normal fertile pollen 8 months after successful transfer into soil. These plants had normal chromosome numbers (2n = 24) but had shorter leaves than original plants. They set seeds after as well as cross pollination.     

Genistein as an endogenous natural substrate of acidic peroxidases in lupin hypocotyls

Crude steam distillate from Ocimum gratissimum sprayed onto infection courts on detached cocoa pods moments after inoculation with Phytophthora palmivora completely inhibited the pathogen and blackpod lesion development on 75% of the infection courts. Disease suppression obtained with the extract was comparable to that obtained with a 2% Kocide 101 suspension. In the field, the O. gratissimum extract also suppressed lesion development although to a significantly lower (P = 0.05) extent in comparison with Kocide 101. Blackpod lesion expansion rates of 3.80, 3.56, 2.71 and 0.78 cm/day, respectively, were associated with pods treated in the field with C. citratus extract, tap water, O. gratissimum extract and 2% Kocide 101. The extract from Cymbopogon citratus was also ineffective on detached pods. Sporangia of P. palmivora from sporu-lating blackpod lesions on both detached and non-detached pods lost their infectivity within 1 h of treatment with the O. gratissimum extract. This effect was superior to that obtained with Kocide 101. Fungitoxicity of the extract on pods, however, was lost within 3 h of application. Thus, despite its in vivo effectiveness as an eradicant, the O. gratissimum extract, in its present form, has limited utility as a protectant fungicide.     

Localization of proteasomes in plant cells

Summary Proteasomes, also known as multicatalytic proteinase complexes, were localized in suspension cells of potato (Solanum tuberosum) by direct immunofluorescence using polyclonal antibodies labelled with fluorescein isothiocyanate. The method used allows an estimate of relative amounts of proteasomal antigens in different cell components. Proteasomes are present in the nuclei and the cytoplasm. The nucleoplasm contains small areas of weak fluorescence. The peripheral cytoplasm and possibly elements of the cytoskeleton show higher fluorescence than other parts of the cytoplasm. This indicates a localization of proteasomes similar to that known from animal cells.Abbreviations DMSO dimethylsulfoxide - EGTA ethyleneglycol-bis-(-aminoethylether)-N,N,N,N-tetra acetic acid - FITC fluorescein isothiocyanate - PBS phosphate buffered saline - PIPES piperazine-1,4-bis-(2-ethanesulfonic acid)     

Dynamic approaches to the mechanism of photosynthesis

An account of the author's life and scientific research is presented. Two main lines of research have been pursued: (1) Studies on the physiological aspect of photosynthesis started from experiments with crops under field conditions and then extended to the study of photosynthesis in nature; and (2) studies on the mechanism of photophosphorylation and related problems which began with the measurement of quantum requirement of photophosphorylation. This work led to the discovery of the high energy state of phosphorylation and many other interesting findings. In recent years, efforts have been made to study the operation and regulation of photosynthetic apparatus with a view to link the above-mentioned lines of research together.Written at the invitation of Govindjee.