Occurrence of beet western yellows virus in sugar beet in Czechoslovakia

The beet western yellows virus (BWYV) was identified in sugar beet plants with leaf yellowing symptoms. When transmitted toSinapis alba L. the virus isolate caused severe symptoms of yellowing and violetting of the interveinal leaf tissue of this plant. By aphidsMyzus persicae (Sulz.) the virus isolate was transmitted toLactuca sativa L.,Raphanus sativus L. var.radicula Pers.,Baphanus sativus L. ssp.sativus L. ap., and toBrassica oleracea L. var.gemmifera DC. InLactuca sativa plants the virus induces a yellowing along with thickenning and brittleness of leaves and with mild dwarfing of the plants. InBaphanus sativus var.radicula andBaphanus sativus ssp.sativus plants it brings about a yellowing of the leaf margins with a change in consistency as was the case in lettuce, and inBrassica oleracea var.gemmifera it causes violet spots on the lower leaf sides. The transmission was proved in repeated experiments by a backtransmission to beet andSinapis alba and further transmission from beet toSinapis alba. The transmission of the virus isolate toVicia faba L.,Datura stramonium L., andPetunia hybrida hort. was unsuccessful. In the course of transmissions the isolate properties did not change. In its host range the virus resembles the Duffus’ strain 3 BWYV, isolated from beet in the U.S.A. This is the first characteristic of an Europian BWYV isolate, as obtained from naturally infected beet plants.     

Studies of the gene expression in a somatic hybrid of dihaploid Solanum tuberosum and diploid S. brevidens using two-dimensional protein electrophoresis

Two-dimensional protein electrophoretic patterns of leaf, stem and microtuber were compared between a somatic hybrid (Solanum tuberosum + S. brevidens) and parental plants. Polypeptide spots observed in leaf of the somatic hybrid (BT-1) were similar to those of S. brevidens. In the stem of BT-1, the spots characteristic for each parental plants were also observed. Three specific spots (W, X, Y) found in BT-1 were identical to those of S. tuberosum, however their appearance in S. brevidens depended on the culture conditions (observed at 16h daylength regime, but not in the dark with high sucrose concentration). Potato tuber storage protein patatin was observed in small amounts in the microtubers of BT-1. The data indicated that gene expression unique to each parental plants also existed in the somatic hybrid.Abbreviations BAP 6-benzylaminopurine - 2,4-D (2,4-dichlorophenoxy) acetic acid - IAA indole-3-acetic acid - MES 2-(N-morpholino) ethanesulfonic acid - NAA 1-naphthaleneacetic acid - PAGE polyacrylamide gel electrophoresis - SDS sodium dodecylsulfate     

Studies on broccoli necrotic yellows virus

Electron microscopy of infected D. stramonium cells showed that the virus particles occurred in the cytoplasm. Particles were mostly bacilliform and measured 297 ± 18 times 64 ± 4 nm. In negatively stained leaf homogenates, particles were mostly disrupted; intact particles measured 267 ± 20 times 69 ± 6 nm. In brussels-sprout cells containing BNYV and CIMV, BNYV particles were rarely found compared with those of CIMV, and they occurred within and near CIMV inclusion bodies. BNYV particles were also found in extracts of virus-carrying Brevicoryne brassicae. Broccoli necrotic yellows (BNYV) and cauliflower mosaic (CIMV) viruses occurred together in naturally infected Brussels sprout plants, which showed conspicuous symptoms, and in cauliflower. BNYV was transmitted to and maintained in Datura stramonium and Hyoscyamus niger. It was partially purified from D. stramonium. Using these preparations, from which inhibitor had been removed, BNYV was manually transmitted to cauliflower, causing mild symptoms, and to Brussels sprout, causing a symptomless infection. BNYV also infected Sinapis alba but not cabbage, lettuce, Sonchus oleraceus or Plantago major. BNYV was transmitted by Brevicoryne brassicae but not by Myzus persicae, Hyperomyzus lactucae or Aleyrodes proletella.     

Growth and tropane alkaloid production inAgrobacterium transformed roots and derived callus ofDatura

Small callus pieces excised from theAgrobacterium transformed root line D₂ ofDatura stramonium, were cultured onto solidified MS medium supplemented with a 1.0 μM kinetin and three different concentrations (0.1, 0.5 and 1.0 μM) of 2,4-dichlorophenoxyacetic acid (2,4-D), and were examined for their alkaloid productivity in relation to organization level and growth rate. Growth of transformed roots (in a MS liquid medium without plant growth regulators) was greater than that of transformed calli excised from them and cultured separately. The addition of 1.0 μM 2,4-D to the culture medium had a positive effect on callus biomass production, while it inhibited root formation by this tissue (the lower the 2,4-D concentration in the medium the greater the number of roots which emerged from the calli). Hyoscyamine production was also higher in the transformed roots than in the transformed calli, and in these tissues the production of hyoscyamine was positively correlated with organogenesis index (i.e. its ability for rooting). At the same time, the epoxidation of hyoscyamine to scopolamine only took place in the transformed calli. This occurred to a greater extent at the lower concentrations of 2,4-D in the culture medium. The mode through which the 2,4-D could control the alkaloid production of transformed callus is discussed.     

Defensive role of leaf trichomes in resistance to herbivorous insects in Datura stramonium

This study assessed the role of leaf trichome density as a component of resistance to herbivores, in six populations of Datura stramonium. Phenotypic selection on plant resistance was estimated for each population. A common garden experiment was carried out to determine if population differences in leaf trichome density are genetically based. Among population differences in leaf trichome density, relative resistance and fitness were found. Leaf trichome density was strongly positively correlated to resistance across populations. In 5 out of 6 populations, trichome density was related to resistance, and positive directional selection on resistance to herbivores was detected in three populations. Differences among populations in mean leaf trichome density in the common garden suggest genetic differentiation for this character in Datura stramonium. The results are considered in the light of the adaptive role of leaf trichomes as a component of defence to herbivores, and variable selection among populations.     

Somatic hybrids of Datura innoxia Mill.+Datura discolor Bernh. and of Datura innoxia Mill.+Datura stramonium L. var tatula L.

Summary Following fusion of protoplasts from a chlorophyll-deficient diploid mutant of Datura innoxia Mill. which can be regenerated to shoots, with green wild-type protoplasts of Datura stramonium L. var. tatula L. which can not, it was possible to isolate 49 green hybrid calli on agar medium. Most of these somatic hybrid calli gave rise to leaves and shoots. The chromosome numbers of the somatic hybrids were determined: 15 were tetraploid (amphidiploid), 24 hexaploid, and the other showed an aneuploid chromosome number.In a similar experiment protoplasts of the Datura innoxia mutant were fused with green wild-type protoplasts of Datura discolor Bernh. which are also not able to be regenerated, four green calli were obtained from which leaves and shoots developed after some transfers on agar medium. Three of them showed the amphidiploid (48) chromosome number, whereas one possessed an aneuploid number of 46 chromosomes.After transfer of rooted shoots to soil flowering plants could be obtained in both combinations. The habits of the somatic hybrids in both combinations were intermediate between the habits of the respective parental plants.Dedicated to my father, Prof. Dr. Theodor Schieder, on the occasion of his 70th birthday.     

Geographic association and temporal variation of chemical and physical defense and leaf damage in Datura stramonium

The evolution of plant defense traits has traditionally been explained trough the “coevolutionary arms race” between plants and herbivores. According to this, specialist herbivores have evolved to cope effectively with the defensive traits of their host plants and may even use them as a cue for host location. We analyzed the geographic association between leaf trichomes, two tropane alkaloids (putative resistance traits), and leaf damage by herbivores in 28 populations of Datura stramonium in central Mexico. Since the specialist leaf beetles Epitrix parvula and Lema trilineata are the main herbivores of D. stramonium in central Mexico, we predicted a positive association between plant defense and leaf damage across populations. Also, if physical environmental conditions (temperature or precipitation) constrain the expression of plant defense, then the geographic variation in leaf damage should be explained partially by the interaction between defensive traits and environmental factors. Furthermore, we studied the temporal and spatial variation in leaf trichome density and leaf damage in five selected populations of D. stramonium sampled in two periods (1997 vs. 2007). We found a positive association between leaf trichomes density and atropine concentration with leaf damage across populations. The interaction between defensive traits and water availability in each locality had a significant effect on the geographic variation in leaf damage. Differences among populations in leaf trichome density are maintained over time. Our results indicate that local plant–herbivore interaction plays an important role in shaping the geographic and temporal variation in plant defense in D. stramonium.     

Outdoor infrared imaging for spatial and temporal thermography: A case study of necrotic versus healthy leaf areas on woody plants

The temperature of plants can be measured using infrared (IR) thermography. Despite the extensive use of IR imaging indoors, outdoor IR imaging is uncommon. We used IR imaging to compare leaf temperatures between necrotic spots and healthy areas of oriental cherry (Prunus serrulata var. spontanea), Japanese cornel (Cornus officinalis) and sawtooth oak (Quercus acutissima) in the field. There was a significant difference in the mean leaf temperatures between the necrotic spots (26.4°C) and healthy areas (25.6°C) of oriental cherry (p = .01). The mean temperatures in the necrotic spots of Japanese cornel and sawtooth oak leaves were 22.3°C and 29.6°C, respectively, which were not significantly different from the mean temperatures of the healthy areas. A consecutive, 2‐day temporal leaf analysis in October 2018 revealed that the temperatures in the necrotic spots were generally higher than those in the healthy areas of the three species. The temperature difference between the spots and healthy areas (up to 1.4°C) was more pronounced at 13:00 hr in all three species on both days. These results reveal differences in the spatial and temporal thermal state across the necrotic spotted leaves. There is potential for use of outdoor IR imaging to visualize the response of trees to pathogen infection and abiotic stress.     

Influence of Two Phenoxy Growth Regulators on the Uptake and Accumulation of Naptalam by Bean Plants

Bean plants (Phaseolus vulgaris L. cv. Black Valentine) treated with 2,4-dichlorophenoxyacetic acid (2,4-D) and 4-chlorophenoxyacetic acid (PCPA) absorb and accumulate considerably more N-1-naphthylphthalamic acid (naptalam) than untreated plants. All concentrations of 2,4-D from 5 x 10^-8 to 5 x 10^-4M were effective, peak stimulation occurring at 3 x 10^-5M. Plants treated with this concentration took up 186% more naptalam than control plants. It was shown that the leaf area (on a per g dry weight basis) was influenced most by growth regulator treatment. The leaf area of plants treated with 5 x 10^-5M 2,4-D contained 575% more naptalam than the leaf area of untreated plants. The influence of PCPA on naptalam uptake by bean plants was similar to that of 2,4-D but less effective.     

Detection of hemicelluloses specific to the cell wall of tracheary elements and phloem cells by fluorescein-conjugated lectins

Summary Binding of fluorescein-conjugated wheat-germ agglutinin (F-WGA) and some other lectins to tissues from various plants were examined by epifluorescence microscopy. F-WGA bound specifically to the walls of tracheary elements (TEs) and phloem cells of pea roots. The binding sites in TEs were localized only in the secondary thickening and became evident at very early stages of differentiation. Fluorescein-conjugated derivatives ofSolanum tuberosum lectin,Lycopersicon esculentum lectin, andDatura stramonium lectin, which bind N-acetylglucosamine residues as WGA, also bound to the secondary thickening of TEs of pea roots. The binding sites for F-WGA were not removed by extraction with hot EDTA and proteinase K, but removed by extraction with an alkali solution. The alkali-extracted binding sites from the roots were precipitated together with hemicelluloses by 80% ethanol. These results indicate that the binding sites are not present on pectins, proteins, or cellulose, but hemicelluloses. Localized distribution of the binding sites for F-WGA in TEs was found also in a variety of angiosperm plants.Abbreviations BSL-II Bandeiraea simplicifolia lectin II - DSL Datura stramonium lectin - F fluorescein-conjugated - LEL Lycopersicon esculentum lectin - MT microtubule - STL Solanum tuberosum lectin - TE tracheary element - WGA wheat-germ agglutinin     

Growth and isoflavonoid accumulation of Pueraria candollei var. candollei and P. candollei var. mirifica cell suspension cultures

We established cell suspension cultures derived from leaf, stem, and root calli of Pueraria candollei var. candollei and P. candollei var. mirifica using liquid Murashige and Skoog (MS) medium supplemented with 0.56 μM 6-benzyladenine (BA) and 4.52 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Growth of the cell suspension cultures progressed to the stationary phase within 15–24 days. Methanolic extracts of cell suspension cultures of both varieties of P. candollei were analyzed using a validated HPLC protocol. All cell lines derived from leaf, stem, and root explants produced four major isoflavonoids: daidzein, daidzin, genistein, and genistin; these isoflavonoids were detected only in the roots of intact plants. Furthermore, the isoflavonoid contents of the cell suspension cultures were higher than those of intact plants. Thus, cell suspension culture of both varieties of P. candollei may be an effective tool for isoflavonoid production.     

Embryogenesis in suspension cultures of Datura innoxia Mill.

In vitro plant regeneration via embryogenesis was obtained in suspension cultures of Datura innoxia Mill. Embryogenesis was induced in suspension cultures raised from callus of androgenetic origin, using LS liquid medium supplemented with 0.22 mg/l 2,4-D. The total number of embryos formed was variable over time in culture. Embryos differentiated and matured in the liquid medium itself as also evidenced by histological observation. Embryos germinated to form plantlets on semisolid MS medium without growth substances. The regenerated plants had haploid number of chromosomes.Abbreviations BAP 6-benzylaminopurine - 2,4-D 2,4-dichlorophenoxyacetic acid - Kn Kinetin - LS Linsmaier and Skoog (1965) - MS Murashige and Skoog (1962)     

Growth responses and Hyoscyamine content ofDatura innoxia under the influence of coal-smoke pollution

This study evaluated the impact of pollution on growth responses inDatura innoxia. Coal-smoke emissions were produced by the Badarpur Thermal Power Plant in Delhi, India. At the polluted site, the size of roots and leaves as well as the number of branches and leaves per plant increased, but shoot lengths and leaf areas were lower, compared with control plants. The net photosynthesis rate, stomatal resistance, and the amount of pigments (chlorophyll a, b, and carotenoids) were less in pollution-affected plants, while stomatal conductance and intercellular CO₂ concentration were higher in these plants. Explants from both sites (polluted and non-polluted), grown in vitro on various combinations of auxin (2,4-D, NAA) and cytokinin (BAP, KN), showed the maximum response on a medium containing NAA (0.1 mg L^-1) with BAP (5.0 mg L^-1). Hyoscyamine content was higher in all parts (root, stem, leaf, and regener-ants) of the polluted plants.     

THE EFFECTS OF MECHANICAL STIMULATION AND ETIOLATION ON THE COLLENCHYMA OF DATURA STRAMONIUM

Walker , Waldo S. (Grinnell College, Grinnell, Iowa.) The effects of mechanical stimulation and etiolation on the collenchyma of Datura stramonium. Amer. Jour. Bot. 47(9) : 717–724. Illus. 1960.–In an effort to determine the effect of mechanical stimulation on collenchyma tissue, plants of Datura stramonium L. were placed on a mechanical agitator and subjected to intensive shaking for 9 hr. per day for 40 days. Measurements indicated that such stimulation greatly increased the amount of wall thickening per cell, as observed in transverse section. Measurements also indicated that such stimulation may inhibit collenchyma cell elongation. A second group of Datura stramonium plants was placed in total darkness to determine the effect of such treatment on the quantity of wall thickening in the collenchyma tissue. Measurements indicated that when plants were placed in the dark for extended periods a great reduction of wall thickening resulted. It is suggested that reduction of wall material was due to its utilization as substrate for respiratory processes which occur in the plant under such extreme conditions. The composition and structure of the collenchyma cell walls are discussed.     

Translocation and Complex Formation of Picloram and 2,4-D in Rape and Sunflower

Uptake, translocation and complex formation of ¹⁴C-labelled 4-amino-3,5,6-trichloropicolinic acid (picloram) and 2,4-dichlorophenoxyacetic acid (2,4-D) in seedlings of rape (Brassica napus L. cv. Nilla) and sunflower (Helianthus annuus L. var. uniflorus) were studied. Sunflower is susceptible both to 2,4-D and picloram, while rape is susceptible to 2,4-D but more tolerant to picloram. The uptake of the herbicides through the leaves was almost complete in both species. Translocation of 2,4-D into the roots took place more readily than that of picloram. In sunflower about 50 per cent of the applied 2,4-D was extruded through the roots into the nutrient solution after 9 days. In the picloram-treated sunflower most of the activity was found in the aerial parts, while in picloram-treated rape most of the activity still occurred in the treated leaf after 9 days. No activity at all was found in the roots or in the nutrient solution of the picloram-treated rape seedlings. While the major part of 2,4-D always was found in the state of free herbicide, a large fraction of picloram was rapidly bound into water-soluble complexes. This binding was especially pronounced in rape. Separation by paper chromatography showed that different radioactive compounds were formed. Most of these could be hydrolyzed, thereby releasing free herbicide. The results support the hypotheses that complex formation could counteract herbicide translocation and toxicity of auxin herbicides.     

Activité néformatrice d'acides ribonucléiques extraits de tumeurs végétales

RNAs extracted from crown-gall tumors, induced by Agrobacterium tumefaciens (strain B6) on stems of Datura stramonium L., have been isolated by the phenol method and purified through Biogel P 60 columns. These RNAs have been transformed into complexes with l -amino acids by incubation in a medium containing: Tris HCI buffer, pH 7.6, ATP, MgCl₂, a mixture of l -amino acids and a polypeptide synthetase purified from Alcaligenes faecalis. The inoculation of stems of Datura plants with these complexes induces the development of nodular outgrowths, whereas other complexes made from RNAs isolated from healthy Datura plants, from Agrobacterium tumefaciens, Escherichia coli and Alcaligenes faecalis do not cause any hyperplasia under our experimental conditions. The analysis of the results obtained, supported by histological studies of the outgrowths, suggests that these neoformations should be of tumoral nature.     

Expression of a bacterial gene in transgenic tobacco plants confers resistance to the herbicide 2,4-dichlorophenoxyacetic acid

Plants resistant to the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) were produced through the genetic engineering of a novel detoxification pathway into the cells of a species normally sensitive to 2,4-D. We cloned the gene for 2,4-D monooxygenase, the first enzyme in the plasmid-encoded 2,4-D degradative pathway of the bacterium Alcaligenes eutrophus, into a cauliflower mosaic virus 35S promoter expression vector and introduced it into tobacco plants by Agrobacterium-mediated transformation. Transgenic tobacco plants expressing the highest levels of the monooxygenase enzyme exhibited increased tolerance to 2,4-D in leaf disc and seed germination assays, and young plants survived spraying with levels of herbicide up to eight times the usual field application rate. The introduction of the gene for 2,4-D monooxygenase into broad-leaved crop plants, such as cotton, should eventually allow 2,4-D to be used as an inexpensive post-emergence herbicide on economically important dicot crops.     

Isolation and characterization of three new PGPR and their effects on the growth of Arabidopsis and Datura plants

This study characterizes three bacterial strains isolated from plant rhizospheres and evaluates their performance as plant-growth-promoting rhizobacteria. Pseudomonas plecoglossicida strain Pp20 was isolated from the rhizosphere of a date palm in Bechar (Algerian Sahara), Bacillus spec. strain Bt04 isolated from the rhizosphere of pear in Ghardaia (Algerian Sahara) and Lysinibacillus fusiformis strain Lf89 was isolated from the rhizosphere of tomato in Ain Defla (northern Algeria). Their effects on plant growth and development were analyzed in different in vitro cultures: an Arabidopsis thaliana plate assay and two hydroponic systems for Datura stramonium and Datura tatula. Our results show that all strains significantly improve plant growth of the plant species tested and some strains produce a shift in the C/N ratio in A. thaliana. Inoculation had no effect on alkaloid production per gram leaf dry weight in D. stramonium, but specific plant-growth-promoting rhizobacteria interactions may alter the alkaloid composition in the shoot.     

Action de l'acide 2,4 dichlorophenoxyacetique (2,4-D) sur la migration du42K fourni par voie foliaire chez la laitue et le maïs

The 2,4 dichlorophenoxyacetic acid (2,4-D) applied in solution (10μg per plant) on a leaf of lettuce plants (Lactuca sativa, cv. Grosse Blonde Paresseuse) decrease the translocation of⁴²K from the treated leaf, both in pretreatment and in simultaneous action if the interval of time is sufficiently long between the foliar applications. The metabolic action of 2,4-D seems to be located in the treated leaf. The results may be explained by a stimulation of the metabolic activity and consequently of the retention, already observed by several authors, with 2,4-D and other phytohormones. The 2,4-D had not a comparable effect on translocation of⁴²K from the leaves of maize plants (Zea mays Dekalb 202) even if the amount applied is higher and the duration of the pretreatment more important than previously. However, it must be noted for this species a light stimulation with 2,4-D on the retention of⁴²K in the sheath of the treated leaf. The results obtained with lettuce and maize might be fitted in a pattern of selectivity which appears for other aspects of the metabolism.     

Response of mesquite to nitrate and salinity in a simulated phreatic environment: Water use,dry matter and mineral nutrient accumulation

Mesquite plants (Prosopis glandulosa var. Torreyana) were grown in 2-m long columns 20 cm in diameter, and provided with a constant, stable ground water source 10 cm above the sealed base of the column. Ground water contained 0, 1 or 5 mM nitrate, or a mixed salt solution (1.4, 2.8, or 5.6 dS m^-1) with the ionic ratios of ground water found in a field stand of Prosopis at Harper's Well (2.8 dS m^-1). Water uptake in the highly salinized columns began to decrease relative to low salt columns when soil salinity probes 30 cm above the column base read approximately 28 dS m^-1, a potential threshold for mesquite salt tolerance. Prosopis growth increased with increasing nitrate, and decreased with increasing salinity. Water use efficiency was little affected by treatment, averaging approximately 2 g dry matter L^-1 water used. Most fine roots were recovered from a zone about 25 cm above the ground water surface where water content and aeration appeared to be optimal for root growth. Root-shoot ratio was little affected by nitrate, but increased slightly with increasing salinity. Plant tissue P concentrations tended to increase with increasing salinity and decrease with increasing N, approaching potentially deficient foliage concentrations at 5 mM nitrate. The whole-plant leaf samples increased in sodium concentration both with added salt and with added nitrate. Foliar manganese concentrations increased with increasing salt in the absence of nitrate. Concentrations of sodium in leaves were low (<10 g kg^-1), considering the high salt concentrations in the ground water. Prosopis appears to exclude sodium very effectively, especially from its younger leaves. Although Prosopis is highly salt tolerant, the degree to which it utilizes soil nitrate in place of biologically fixed N may lower its salinity tolerance and affect its nutrient relations in phreatic environments.