APOMIXIS AND POLYEMBRYONY IN HIEROCHLOË ODORATA

Norstog , Knut . (Wittenberg U., Springfield, Ohio.) Apomixis and polyembryony in Hierochloe odorata. Amer. Jour. Bot. 50(8): 815–821. Illus. 1963.—Hierochloë odorata from Michigan, having 2n = 56 chromosomes, was found to reproduce by pseudogamous fertilization of diploid aposporous embryo sacs. Diploid embryos, 2n= 56, and 5n = 140 ± endosperm occurred together. Megasporogenesis was incomplete, and aposporous embryo sac initials developed directly into 8-nucleate Polygonum type embryo sacs. Microsporogenesis was irregular with univalent, bivalent and multivalent chromosomes in meiosis-I. Dyads and microspores varied between n = 24 and n = 32, and less than 50% of the pollen grains stained with acetocarmine. Two other races of H. odorata are known to occur in North America. They are an apparently infertile type in Canada with 2n = 28, and a perfectly fertile race from coastal Connecticut also with 2n = 28. It is suggested that the H. odorata with 2n = 56 is a derivative of the sterile Canadian race.     

Direct Haplotype‐Specific DNA Sequencing

Abstract

Determining haplotype‐specific DNA sequence information is very important in a wide range of research fields. However, no simple and robust approaches are currently available for determining haplotype‐specific sequence information. We have addressed this problem by developing a very simple and robust haplotype‐specific sequencing approach. We utilise the fact that DNA sequencing polymerases are sensitive to 3′end mismatches in the sequencing primer. By using two sequencing primers with 3′end corresponding to the two alleles in a given SNP locus, we are able to obtain allele‐specific DNA sequences from both alleles.

We evaluated this direct haplotype‐specific approach by determining haplotypes within the intron 2 sequence of the fructan‐6‐fructosyltransferase (6‐ft) gene in Lolium perenne L. We obtained reliable haplotype‐specific sequences for all primers and genotypes evaluated. We conclude that the haplotype‐specific sequencing is robust, and that the approach has a potentially very wide application range for any diploid organism.     

Centrosomal ALIX regulates mitotic spindle orientation by modulating astral microtubule dynamics

The orientation of the mitotic spindle (MS) is tightly regulated, but the molecular mechanisms are incompletely understood. Here we report a novel role for the multifunctional adaptor protein ALG‐2‐interacting protein X (ALIX) in regulating MS orientation in addition to its well‐established role in cytokinesis. We show that ALIX is recruited to the pericentriolar material (PCM) of the centrosomes and promotes correct orientation of the MS in asymmetrically dividing Drosophila stem cells and epithelial cells, and symmetrically dividing Drosophila and human epithelial cells. ALIX‐deprived cells display defective formation of astral microtubules (MTs), which results in abnormal MS orientation. Specifically, ALIX is recruited to the PCM via Drosophila Spindle defective 2 (DSpd‐2)/Cep192, where ALIX promotes accumulation of γ‐tubulin and thus facilitates efficient nucleation of astral MTs. In addition, ALIX promotes MT stability by recruiting microtubule‐associated protein 1S (MAP1S), which stabilizes newly formed MTs. Altogether, our results demonstrate a novel evolutionarily conserved role of ALIX in providing robustness to the orientation of the MS by promoting astral MT formation during asymmetric and symmetric cell division.     

Experimental Lake Fertilization in the Kuokkel Area,Northern Sweden. Phytoplankton Biomass and Algal Composition in Natural and Fertilized Subarctic Lakes

Based on 500 quantitative algal analyses from 1971–1980 the seasonal variations of phytoplankton biomass and species diversity have been evaluated in one natural and three fertilized subarctic lakes. Enrichment with P alone did not cause any biomass increase and inhibited dinoflagellates and Dinobryon. Enrichment with N alone stimulated Uroglena and Gymnodinium and the biomass increased. No species were inhibited. Enrichment with both N and P made the biomass increase 50–60 times and the stimulated genera were: Chlorella, Choricystis, Chromulina, Chrysochromulina, Gymnodinium, Monoraphidium, Ochromonas, Spiniferomonas, Rhodomonas and Uroglena. In all fertilized lakes monocultures of chlorococcal green algae finally developed.     

Cell arrangement,mass and dimension ofStreptococcus faecalis during growth

During exponential growth ofStreptococcus faecalis, the distribution of cell arrangements remains constant, but depends on the growth rate. The predominant cell arrangements are diplococci (about 60–80% of total cells) the amount of which varies only little with the growth rate. A clear correlation exists for cells growing as chains; the amount decreases from about 20% at μ=2.0 to about 6% at μ=0.45. After cessation of growth in the stationary phase, the number of diplococci and chains decreases and the number of monococci increases; after 10 h in the stationary phase, more than 50% of the cells have become monococci. The dry weight of 2.5×10⁻¹⁰ mg/cell remains constant at different growth rates, while cell size shows small differences on different growth media. Treatment of exponentially growing cultures with crystal violet or nitrofurantoin results in faster sedimentation on sucrose gradients of treated cultures compared to untreated cultures. While crystal violet effects an increased chain formation, treatment with nitrofurantoin results in an increase of the size of the individual cell.     

Superstoichiometric binding of L-Phe to phenylalanine hydroxylase from Caenorhabditis elegans: evolutionary implications

Phenylalanine hydroxylase (PAH) catalyzes the hydroxylation of L-Phe to L-Tyr. Dysfunctional PAH results in phenylketonuria and mammalian PAH is therefore highly regulated and displays positive cooperativity for L-Phe (Hill coefficient (h) = 2). L-Phe does not bind to the regulatory ACT domain in full-length tetrameric human PAH and cooperativity is elicited by homotropic binding to the catalytic site (Thórólfsson et al. in Biochemistry 41:7573–7585, 2002). PAH from Caenorhabditis elegans (cePAH) is devoid of cooperativity for L-Phe (h = 0.9), and, as shown in this work, structural analysis reveal an additional L-Phe binding site at the regulatory domain of full-length cePAH. This site involves the GA(S)L/ISRP motifs, which are also found in ACT domains of other L-Phe binding proteins, such as prephenate dehydratase. Isothermal titration calorimetry further demonstrated 2 binding sites per subunit for cePAH versus ~1 for hPAH. Steric occlusion of the regulatory site, notably by residues Lys215/Tyr216 from the adjacent catalytic domain, appears to hinder regulatory binding in full-length hPAH. Accordingly, the humanized mutant Q215K/N216Y of cePAH binds ~1.4 L-Phe/subunit. This mutant also displays high catalytic activity and certain positive cooperativity for L-Phe (h = 1.4). Our results support that the acquisition of positive cooperativity in mammalian forms of PAH is accompanied by a closure of the regulatory L-Phe binding site. Concomitantly, the function of the regulatory ACT domain appears to be adapted from amino acid binding to serving the communication of conformational changes among catalytic subunits.     

Evaluation of PM3(tm) as a Geometry Generator in Theoretical Studies of Transition-Metal-Based Catalysts for Polymerizing Olefins

The PM3(tm) method has been applied to several systems of relevance to catalytic polymerization of olefins, for catalysts containing Ti, Zr or Cr. With some exceptions, PM3(tm) calculations reproduce experimental geometries of stable, closed-shell, precursors well. For stationary points along the path of monomer insertion into a metal-alkyl bond, the comparison is made to structures obtained by optimization using various first-principle methods. Large errors are uncovered for the transient structures, in particular pertaining to metal-ethylene coordination and agostic interactions. The energy profiles for four insertion reactions are computed by gradient-corrected density functional (DFTG) methods, using molecular structures taken from PM3(tm) and first-principle geometry optimizations, respectively. The chromium case is promising, giving values for the barrier to monomer insertion of 11 and 9 kcal/mol based on PM3(tm) and DFTG geometries, respectively. The Ti- and Zr-based systems are predicted to proceed downhill based on PM3(tm) structures, whereas small barriers are found when using first-principle structures. A hybrid PM3(tm)-DFTG procedure is suggested for geometry optimization, which facilitates an accurate estimate of the barrier when applied to one of the zirconium systems.