The role of ethylene in the control of cell division in cultured pea root tips: a mechanism to explain the excision effect

Summary A transitory cell division block, or excision effect, occurs in the meristem of roots after excision and transfer to culture medium. This block can be induced, in intact seedling roots, by exogenous treatment with ethylene gas. With continuous treatment, the block is longer and the recovery less than after a 4 hour pulse. In excised roots the excision effect can be eliminated by treatment with an inhibitor of ethylene synthesis (aminoethoxyvinylglycine) or action (silver thiosulfate). These experiments provide evidence to support the hypothesis that ethylene from the wounded end of an excised root is involved in a process resulting in a transitory block in cell cycle progression in the meristem. The implications of this hypothesis are discussed.     

Growth in vitro of arrested embryos from lethal mutants ofArabidopsis thaliana

Summary Seventeen embryo-lethal mutants ofArabidopsis thaliana with lethal phases ranging from the globular to mature cotyledon stages of development were analyzed by culturing arrested embryos on nutrient media designed to promote either callus formation or the completion of embryo development and the recovery of homozygous mutant plants. Enriched media supplemented with vitamins, amino acids, and nucleosides were used to identify potential auxotrophic mutants. Wild-type embryos produced extensive callus on basal and enriched media supplemented with 2,4-D and kinetin. Numerous roots developed when wildtype callus was grown in the presence of NAA and kinetin. Mutant embryos arrested prior to the heart stage of development formed only a slight amount of callus on basal and enriched media. Arrested embryos from mutants 122G-E and 112A-2A reached a later stage of development and gave the most interesting responses in culture. 122G-E mutant embryos failed to grow on basal media but produced extensive callus and homozygous mutant plants on enriched media. The specific nutrient required for growth of this mutant remains to be determined. Arrested embryos from mutant 112A-2A developed into abnormal plants without roots when placed in culture. Mutant callus also failed to form roots on a variety of root-inducing media. Expression of this mutant gene therefore disrupts development of the root apical meristem during both embryogenesis in vivo and organogenesis in vitro.     

Isolation and fractionation of CHO chromosomes in aqueous two phase systems using charged polymers and base specific macroligands

Summary Chromosomes were isolated in a preparative scale by synchronisation of CHO cells with a double Thymidine block followed by an arrest in the metaphase by addition of Colcemid. Under proper cultivation conditions a mitotic index of 77% total cells could be routinely achieved. Bulk chromosome preparations free of nuclei and other subcellular particles have been obtained by low speed centrifugation followed by a 60 transfer countercurrent distribution using aqueous two phase systems composed of polyethylenglycol and dextran. The partition of CHO chromosomes previously purified in aqueous two phase systems were studied further to develop a protocol for the separation and isolation of individual chromosomes. Partition experiments with chromosomes changing the electrostatic phase potential by addition of charged PEG-derivatives suggest the existence of relatively highly charged chromosome groups. Most promising results with regard to separation were obtained using two PEG-derivatives, which interact specifically with the bases in DNA. For this affinity partitioning a GC- and AT-specific macroligand were employed. Comparing CCD's using each of these ligands information on the GC and AT content of exposed DNA in the chromosomes groups could be derived, demonstrating that specific sequences of DNA are accessible at the surface of metaphase chromosomes.     

Symbiotically defective histidine auxotrophs of Bradyrhizobium japonicum

Four histidine auxotrophs of Bradyrhizobium japonicum strain USDA 122 were isolated by random transposon Tn5 mutagenesis. These mutants arose from different, single transposition events as shown by the comparison of EcoRI and XhoI-generated Tn5 flanking sequences of genomic DNA. The mutants grew on minimal medium supplemented with l-histidine or l-histidinol but failed to grow with l-histidinol phosphate. While two of the muants were symbiotically defective and did not form nodules on Glycine max cvs. Lee and Peking and on Glycine soja, the other two mutants were symbiotically competent. Reversion to prototrophy occurred at a frequency of about 10^-7 on growth medium without added antibiotics, but prototrophs could not be isolated from growth medium containing 200 g/ml kanamycin and streptomycin. The prototrophic revertants formed nodules on all the soybean cultivars examined. When histidine was supplied to the plant growth medium, both nodulation deficient mutants formed effective symbioses. On histidine unamended plants, nodules were observed infrequently. Three classes of bacterial colonies were isolated from such infrequent nodules: class 1 were kanamycin resistant-auxotrophs; class 2 were kanamycin sensitive-prototrophs; and class 3 were kanamycin-sensitive auxotrophs. Our results suggest that two Tn5 insertion mutations in B. japonicum leading to histidine auxotrophy, affect nodulation in some way. These mutations are in regions that show no homology to the Rhizobium meliloti common nodulation genes.     

Correlation between infection by Rhizobium leguminosarum and lectin on the surface of Pisum sativum L. roots

The lectin on the surface of 4- and 5-dold pea roots was located by the use of indirect immunofluorescence. Specific antibodies raised in rabbits against pea seed isolectin 2, which crossreact with root lectins, were used as primary immunoglobulins and were visualized with fluorescein- or tetramethylrhodamine-isothiocyanate-labeled goat antirabbit immunoglobulin G. Lectin was observed on the tips of newly formed, growing root hairs and on epidermal cells located just below the young hairs. On both types of cells, lectin was concentrated in dense small patches rather than uniformly distributed. Lectin-positive young hairs were grouped opposite the (proto)xylematic poles. Older but still-elongating root hairs presented only traces of lectin or none at all. A similar pattern of distribution was found in different pea cultivars, as well as in a supernodulating and a non-nodulating pea mutant. Growth in a nitrate concentration which inhibits nodulation did not affect lectin distribution on the surface of pea roots of this age. We tested whether or not the root zones where lectin was observed were susceptible to infection by Rhizobium leguminosarum. When low inoculum doses (consisting of less than 10⁶ bacteria·ml^-1) were placed next to lectin-positive epidermal cells and on newly formed root hairs, nodules on the primary roots were formed in 73% and 90% of the plants, respectively. Only a few plants showed primary root nodulation when the inoculum was placed on the root zone where lectin was scarce or absent. These results show that lectin is present at those sites on the pea root that are susceptible to infection by the bacterial symbiont.Abbreviations FITC fluorescein isothiocyanate - TRIC tetramethylrhodamine isothiocyanate     

甘蔗伤流液中保绿活性物质的分离与初步鉴定

通过化学与物理学方法,从甘蔗(Sugar-cane)伤流液中分离出五种活性因素,对离体叶片有不同程度的保绿效应。依其化学性质判断,应属嘌呤化合物,可能是细胞分裂素物质。     

Isolation of a brain peptide identical to the intestinal PHI (peptide HI)

The isolation of a brain peptide identical to the intestinal peptide PHI (peptide HI) is described. The peptide was isolated from porcine brain extract using a chemical assay method based on its C-terminal isoleucine amide structure. The complete amino acid sequence of the peptide was found to be: His-Ala-Asp-Gly-Val-Phe-Thr-Ser-Asp-Phe-Ser-Arg-Leu-Leu-Gly-Gln-Leu-Ser-Ala- Lys-Lys-Tyr-Leu-Glu-Ser-Leu-Ile-NH2. This sequence is identical to the intestinal peptide thus demonstrating PHI to be a brain-gut peptide. The role of PHI in the central nervous system as a neurotransmitter or neuromodulator is discussed.     

Root development and configuration in intensively managed radiata pine plantations

Summary In south-east Australia, where radiata pine (Pinus radiata D. Don) is grown on sandy soils low in nutrients and short of water, early establishment, and rapid growth to canopy closure lead to increased productivity. At this stage demands for nutrients and water are high, and trees respond vigorously to silvicultural inputs.For several months after transplanting in winter roots are confined within a narrow planting wedge, low temperature restricts new root growth and slows recovery from water stress in plants. From spring, depending upon the configuration and vigour of the roots transplanted, lateral roots extend radially throughout the soil.Although there were small decreases in concentration of roots radially from the stems of very young trees, such spatial differences disappeared between ages 2 and 3, so that rooting density was independent of distance from the stem. The pattern of vertical distribution of lateral roots was not influenced by age and 80–90% of the lateral roots were within the top 30 cm soil. Roots developed rapidly as the trees grew towards canopy closure, but in general the rooting densities of these pines are among the lowest reported for plants. In rapidly growing trees approaching canopy closure, the secondary thickening of the lateral roots was sufficient to double the weight of roots without altering root length.Knowledge about root growth and root configuration during the early phase of plantation development will assist management decisions where intensive silviculture is practiced, and hence ensure the most efficient use of nutrients and water.     

Root growth and litter decomposition in a coffee plantation under shade trees

In a non-fertilized coffee plantation under shade trees the root biomass was excavated to estimate its distribution in the soil profile. One third of total fine (less than 1 mm) roots was found in the first 10 cm of soil; the cumulative total to 30 cm reached 73%. A highly variable and transient amount of fine roots colonized the litter layer. Root production both in the litter and in the first 7.5 cm of mineral soil was estimated from sequential samplings and was 10 g m^–2 yr^–1 and 660 g m^–2 yr^–1 respectively. The decomposition rate of weighed averages of litter fractions in the coffee plantation, calculated as the ratio of litter fall rate to the amount found in the soil was k=4.8. Shade tree leaves, the major component of litter descomposed slower than coffee leaves and these slower than flowers and fruits. Litter bag experiments showed considerable slower rates when mesh was 0.03 mm than 0.5 mm. Nitrogen and phosphorous showed increases in concentrations as decomposition progressed while potassium, calcium and magnesium followed a decrease in concentration that paralleled that of dry weight loss. In comparing the decomposition rate for litter with or without coffee roots growing in the bags, a tendency to show faster decomposition rates was found for the treatment with roots. These differences were however, only significant for one month for shade tree leaves litter. Nitrogen amounts remaining in shade tree leaves litter was lower in the treatment with roots that without roots. Potassium concentration in roots was positively correlated with potassium concentration in decomposing leaf litter where roots were growing. These results suggest that while roots growing attached to decomposing litter had little or no effect in speeding the decomposition process, the superficial roots seem to play an important role in absorbing very efficiently the mineralized nutrients from litter. The anatomical study of roots showed that the plantation is intensely infected with V-A mycorrhiza. External mycorrhizal hyphae did not to play a role in attachment of roots to decomposing litter while root hairs were found to grow in profusion on root surfaces oriented toward litter.     

Effect of temperature on drought resistance and growth of cotton plants

In cotton plants ( Gossypium hirsutum L. cv. B.J.A.) the temperature of the roots affected both root and shoot growth, as did the temperature of the shoot. Drought resistance increased when the temperature imposed on roots (27°C) was lower than that imposed on shoots (17°C); the result was a decrease in both transpiration and flow of root sap. Stomatal characteristics as measured by density, index and resistance, depended only on shoot temperature. Differences in drought resistance, depended only on shoot temperature. Differences in drought resistance seem to be a result of changes in transpiration flow modulated by the amount of absorbed water.