From patterns to clones in chimaeric tissues

This essay summarizes recent advances made in the analysis of mosaic patches in chimaeric epithelia of the mouse. The conclusions drawn from the observed patterns are relevant to the behaviour of expanding cell populations during tissue growth and homeostasis. References are made to clonal analysis of Drosophila.     

Anchoring of rice BAC clones to the rice genetic map in silico

A wealth of molecular resources have been developed for rice genomics, including dense genetic maps, expressed sequence tags (ESTs), yeast artificial chromosome maps, bacterial artificial chromosome (BAC) libraries and BAC end sequence databases. Integration of genetic and physical maps involves labor-intensive empirical experiments. To accelerate the integration of the bacterial clone resources with the genetic map for the International Rice Genome Sequencing Project, we cleaned and filtered the available EST and BAC end sequences for repetitive sequences and then searched all available rice genetic markers with our filtered databases. We identified 418 genetic markers that aligned with at least one BAC end sequence with >95% sequence identity, providing a set of large insert clones with an average separation of 1 Mb that can serve as nucleation points for the sequencing phase of the International Rice Genome Sequencing Project.     

Response of Solidago canadensis clones to competition

Summary Transplants of ten Solidago canadensis clones were grown under high and low competition in the field to determine whether clones differed in survival, growth, and reproduction under natural conditions. Transplants had higher probability of survival and flowering and were larger in all measures of size when competition was experimentally reduced. Clones differed in almost all these measures of success, but only when variance among transplants within clones was reduced by excluding transplants that experienced heavy herbivore damage. Differences among clones were more apparent under low competition than under high competition, despite higher coefficients of variation within clones under low competition. Adjusting transplant size for initial size (parent ramet rhizome mass) did not change these results, although clones did differ in parent rhizome mass. All of these results suggest that there is little potential for selection to discriminate among these clones. Despite the strong differences in transplant performance between the competition treatments and among clones, the clones did not differ in competitive ability-almost none of the clone x competition interactions were significant. In addition, the measures of success of each clone were usually positively correlated between the high and low competition treatments, suggesting there were no tradeoffs between performance under high and low competition for these clones.     

Fertilization of poplar clones in the nursery

Summary Results of experiments with four poplar clones and various chemical fertilizers in a nursery in southern Greece are presented. At the end of the first growth period the heights of the four clones, without fertilizers, decreased in the order of I-214>I-262>cv. campeator > black poplar 1/64 with significant differences only between black poplar 1/64 and the rest of the clones.Of the fertilizer nutrients N, P, K and Mg only N improved heights of all clones significantly and especially of the clone I-214. One hundred and 200 kg of P fertilizer per ha had minimal or negative effect on height increase of all clones.Ammonium sulfate, ammonium nitrate and potassium nitrate all at 400 kg N per ha were found equally effective in improving height growth of the clone I-214 but ammonium nitrate is the N fertilizer of choice by its higher N content and relatively lower price.Ammonium nitrate at 200 kg N per ha, in two or three equal dosages, during the first growth period, June–July, gave the maximum height increase for two consecutive years of the clone I-214. Six hundred kgs, of N per ha reduced height increase of the same clone and increased losses of N, as NO₃ ^–, in drainage water.     

Anti-CD3 antibody induces unresponsiveness to IL-2 in Th1 clones but not in Th2 clones

Culture of murine T cells with immobilized (platebound) anti-CD3 antibody results in autocrine growth factor secretion in both Th1 (IL-2 producing) and Th2 (IL-4 producing) cells. Using a panel of murine T cell clones, we demonstrate that the IL-2-induced proliferation of Th1 clones is dramatically inhibited by immobilized anti-CD3 antibody, whereas that of Th2 clones is not. This unresponsiveness of Th1 clones to IL-2 is not due to decreases in IL-2R expression. Supernatants from Th1 or Th2 cell cultures fail to alter the effects of anti-CD3 on the two types of clones, suggesting that unresponsiveness induced in Th1 clones or the lack thereof in Th2 clones is not mediated by a stable cytokine(s). Accessory cells enhance the proliferation of Th1 cells exposed to low concentrations of anti-CD3, but the unresponsiveness induced by high concentrations of anti-CD3 is not prevented by accessory cells. Finally, soluble anti-CD4 antibody prevents the induction of the unresponsive state even at high concentrations of anti-CD3. These experiments demonstrate that two subsets of cloned CD4+ T cells differ in their responses to anti-CD3, and that CD4 molecules may play a critical role in regulating the outcome of receptor-mediated stimulation.     

Photosynthetic responses to salinity in two clones of Agrostis stolonifera

Abstract The response of photosynthesis, F_e, to salinity was examined in two clones of Agrostis stolonifera, one derived from a salt marsh (SM) and the other from an inland site (IL). The response of young and old leaves was examined over one week of salt treatment and changes in leaf water relations and concentrations of Na⁺, Cl^? and K⁺ ions were also monitored. Sodium⁺ and chloride^? concentrations rose much more sharply in the older leaves of both clones and F_e declined to ca. zero by Day 4. F_e in the young leaves of the SM clone was unaffected by salt whilst the young leaves of the IL clone showed a 20% reduction in F_e within 24 h of salt application. This decline was due to a decline in stomatal conductance and a reduction in the photosynthetic capacity of the mesophyll and was associated with a greater accumulation of Na⁺ and Cl^? ions and loss of K⁺ ions than in the SM clone. The significance of the salt exclusion from the young leaves of the SM clone to its growth on the salt marsh is discussed.     

The use of specific helper T cell clones to study the regulation of isotype expression by antigen-stimulated B cell clones

In order to determine the mechanism by which helper T cells regulate the production of the various immunoglobulin (Ig) classes, a number of helper T cell clones specific for keyhole limpet hemocyanin (KLH) were generated. These helper T cell clones then were used in a modified splenic fragment system whereby cloned helper T cells and a source of B cells were limit-diluted into naive, lethally irradiated recipients. The B cell clones that were subsequently stimulated in such an assay system by the addition of the antigen 2,4-dinitrophenol (DNP)-KLH then were tested for the various isotypes produced. The results of these studies indicate that the use of a single helper T cell clone could result in the production of all known Ig isotypes including IgE. Moreover, the use of a single helper T cell clone could result in multiple isotype production by a single B cell clone. However, a comparison of the isotypes secreted by a number of different B cell clones that were stimulated with the same helper T cell clone indicated that a variety of isotypic patterns could be obtained. In addition, it was found that the majority of B cell clones produced in the presence of T cell clones secrete fewer numbers of different isotypes compared with B cell clones generated with a heterogeneous population of T cells. Finally, no evidence could be found for isotype-specific helper T cell clones, although a few of the T cell clones appeared to induce a somewhat restricted isotype pattern in which only two or three different isotypes were observed.     

Majority of random cDNA clones correspond to single loci in the tomato genome

Summary The number of loci corresponding to each of 34 unique, random cDNA clones has been determined for the diploid plant species Lycopersicon esculentum (tomato) (2n=24). Fifty-three percent of the clones are homologous to loci represented only once in the tomato chromosomes. Thirty-two percent of the clones correspond to two genetically independent loci. The remaining clones belong to gene families represented by 3–5 loci. To determine the number of gene copies per locus, reconstruction experiments were performed on six of the single locus clones. The majority of these loci were estimated to contain only 1 or 2 copies of the gene. These results support the concept that the majority of structural genes in this diploid plant species are arranged in single loci and present in low copy number. Multigene families, such as those for the small subunit of ribulose bisphosphate carboxylase, chlorophyll a/b binding polypeptide and actin are exceptions to this rule.