Studies on G-bands and Macrocoils in Plant Chromosomes

Four different methods including trypsin urea, SDS and NaOH are presented for the in situ induction of G-bands and macrocoils on the chromosomes of Secale cereale, Hordeum vulgare and Vicia faba. The bands obtained were numerous and along the whole chromosome, the number of the G-bands was much interrelated with the condensation of chromosomes. The bands of homologous chromosomes in some cells were matchable. The G-banded chromosomes in late prophase have nearly reached high resolution level. When incubation periods were beyond critical time for G-banding, macrocoils were often revealed. Gyre number changed with chromosome condensation and the direction of coils has showed different patterns. Transformation of G-bands into macrocoils was first reported in plant chromosomes. Some chromosomes showing G-bands under light microscope appeared spiral patterns under scanning electron microscope. In this paper the relationship between G-bands and macrocoils in plant chromosomes is also discussed.     

High-resolution HKG-banding in maize mitotic chromosomes

Root-tip maize chromosomes (2n=20) were prepared by high-resolution procedures after ethidium bromide/colchicine synchronization. Using HKG-banding (HCl−KOH-Giemsa), that shows both centromeric and intercalary heterochromatin, the banding pattern of the elongated-chromosomes showed one to nine well-resolved dark bands. Differences of HKG-banding pattern of elongated and compacted chromosomes were performed by image analysis.     

Microdissection of plant chromosomes by argon-ion laser beam

Summary Rice and barley chromosomal samples were prepared both on a polyester membrane and on an ordinary glass slide and subjected to microdissection by an argon-ion laser. The intensity and the position of the laser beam were controlled by a microcomputer. The most suitable intensity to obtain chromosomal fragments was determined experimentally. As a result, specific regions of the centromere, satellite, short arm, or long arm, of the barley and rice chromosomes were dissected out from the chromosomal spreads. Chromosomal fragments were also successfully transferred from the sample into an Eppendorf tube.     

两种鼠耳蝠的核型、G带和C带研究

使用肺、心脏等组织进行培养,用空气干燥法制作染色体标本,以胰酶法制作G带,BSG法制作c带,分析了贵州2种鼠耳蝠的核型、G带和c带.水鼠耳蝠Myotis daubentoni和小鼠耳蝠Myotis dividii的染色体数均为2n=44,拥有3对大型和1对中型中着丝粒染色体,染色体臂数(FN)=52;这2种鼠耳蝠的G带...     

A comparison of sequence resolution on plant chromosomes: PRINS versus FISH

 The resolution of the chromosomal positions of six high- and one low-copy sequences by oligonucleotide-primed in situ (PRINS) labelling was compared with corresponding data obtained after fluorescent in situ hybridization (FISH) on field-bean and barley chromosomes. While PRINS proved to be suitable for the rapid detection of high-copy tandem repeats at the same loci as those revealed by FISH, no clear PRINS signal was obtained for the low-copy family of vicilin genes at their locus on field-bean chromosome II. This indicates that localization of short target sequences by primer extension via Taq polymerase in situ does not yet provide a resolution equal, or superior, to FISH on plant chromosomes. Therefore, the use of a cocktail of chromosome-specific single-copy sequences as primers for PRINS is no alternative for the not as yet feasible chromosome painting in plants. Received: 21 April 1998 / Accepted: 12 May 1998     

Mapping of repeated DNA sequences in plant chromosomes by PRINS and C-PRINS

 The primed in situ DNA labelling (PRINS) procedure was optimised for the rapid physical mapping of several types of repetitive DNA sequences on the mitotic chromosomes of Vicia faba, Pisum sativum and Secale cereale. A localization of the highly repeated FokI sequence on V. faba chromosomes was achieved after a 7-min total reaction time. In addition, we report a procedure for direct cycling-PRINS (C-PRINS), a variation of PRINS which involves a sequence of thermal cycles analogous to the polymerase chain reaction. Compared to PRINS, C-PRINS was more sensitive. Further work is needed to improve the sensitivity of the reaction to allow for the reliable detection of low-copy DNA sequences. Received: 17 September 1996 / Accepted: 18 October 1996     

Elasticity measurements show the existence of thin rigid cores inside mitotic chromosomes

Chromosome condensation is one of the most critical steps during cell division. However, the structure of condensed mitotic chromosomes is poorly understood. In this paper we describe a new approach based on elasticity measurements for studying the structure of in vitro assembled mitotic chromosomes in Xenopus egg extract. The approach is based on a unique combination of measurements of both longitudinal deformability and bending rigidity of whole chromosomes. By using specially designed micropipettes, the chromosome force-extension curve was determined. Analysis of the curvature fluctuation spectrum allowed for the measurement of chromosome bending ridigity. The relationship between the values of these two parameters is very specific: the measured chromosome flexibility was found to be 2,000 times lower than the flexibility calculated from the experimentally determined Young modulus. This requires the chromosome structure to be formed of one or a few thin rigid elastic axes surrounded by a soft envelope. The properties of these axes are well-described by models developed for the elasticity of titin-like molecules. Additionally, the deformability of in vitro assembled chromosomes was found to be very similar to that of native somatic chromosomes, thus demonstrating the existence of an essentially identical structure.     

Why do adult insects not moult?

Hypotheses are presented concerning why mayflies moult after functional wings develop and why most insects cease to moult at this time. The pattern of retention or loss of the subimaginal moult in extant mayflies suggests that this moult may be necessary to complete elongation of caudal filaments and forelegs of adults. It is then analogous to the pupal moult of holometabolous insects. I propose that selection for wing efficiency has normally confined the presence of functional wings to one instar only, which is also the last and reproductive stage. Selection for light wings has generally caused the epidermis of membranous insect wings to degenerate, thereby precluding otherwise advantageous adult moults.     

Why incongruous knee replacements do not fail early

Rapid failure of knee prostheses does not usually occur, despite the non-conforming nature of the articulation between femoral and tibial components and the associated large contact pressures. This theoretical study examines the likelihood of fatigue fracture of a layered elastic model loaded by a sliding cylindrical indenter. Cracks (line-defects) were assumed to have nucleated within the layer. The stress intensity factors (SIFs) associated with these cracks were calculated. The values obtained for the SIFs are quite low, with a corresponding low likelihood of crack-growth. When taken in conjunction with the experimentally derived fatigue laws of previous investigators, they suggest that short line-cracks should not grow. It seems that early failures have not simply been due to large shear stresses which occur beneath the prosthesis surface. Other factors, such as the degradation of material through heat-pressing, sterilisation or oxidation, or the deleterious effect of fusion defects, may be required to drive the cracks to delamination.     

Why acetylcholinesterase reactivators do not work in butyrylcholinesterase

The pyridinium oxime therapy for treatment of organophosphate poisoning is a well established, but not sufficient method. Recent trends also focus on prophylaxis as a way of preventing even the entrance of organophosphates into the nervous system. One of the possible prophylactic methods is increasing the concentration of butyrylcholinesterase in the blood with the simultaneous administration of butyrylcholinesterase reactivators, when the enzyme is continuously reactivated by oxime. This article summarizes and sets forth the structural differences between butyrylcholinesterase and acetylcholinesterase, essential for the future design of butyrylcholinesterase reactivators. Butyrylcholinesterase lacks the reactivator aromatic binding pocket found in acetylcholinesterase, which is itself a part of the acetylcholinesterase peripheral anionic site. This difference finally renders the current acetylcholinesterase reactivators, when used in butyrylcholinesterase, non-functional.     

Why caterpillars do not grow short and fat

The integument of a caterpillar consists of a cuticle that is structurally isotropic in the plane of the surface, overlaying an epidermis and basal lamina. Between ecdyses the cuticle stretches so that the proportions of caterpillars are about the same at the end of a stadium as at the beginning. This introduces a paradox, since under uniform internal hydrostatic pressure, the hoop stress is twice the axial stress. Stress in the integument of a caterpillar inflated by compressed hemolymph would be twice as great in the circumference as in the axis, causing it to become progressively fatter in proportion to its length as it grows. This does not happen because axial pleats allow caterpillar cuticle to extend more easily in the axis, allowing uniform growth in spite of the greater hoop stress from internal hydrostatic pressure.     

Uptake of isolated plant chromosomes by plant protoplasts

For mass isolation of plant metaphase chromosomes, cultured cells of wheat (Triticum monococcum) and parsley (Petroselinum hortense) were synchronized by hydroxyurea and colchicine treatment. This synchronization procedure resulted in high mitotic synchrony, especially in suspension cultures of parsley in which 80% of the cells were found to be at the metaphase stage. Mitotic protoplasts isolated from these synchronized cell cultures served as a source for isolation of chromosomes. The described isolation and purification method yielded relatively pure chromosome suspension. The uptake of the isolated plant chromosomes into recipient wheat, parsley, and maize protoplasts was induced by polyethylene-glycol treatment. Cytological studies provided evidences for uptake of plant chromosomes into plant protoplasts.Abbreviations PEG polyethylene glycol - HU hydroxyruea - C colchicine - HUC hydroxyurea and colchicine - CIM chromosome isolation medium - TCM Tris chromosome medium     

In situ fluorescent nick translation procedure for plant chromosomes

Pectinase and cellulase, which are used to macerate plant material, always show traces of DNase activities that result in DNA nicking. Moreover, the DNA polymerase I usually applied in the in situ nick translation techniques shows both 5' to 3' and 3' to 5' exonuclease activities. As a result, significant nonspecific labeling appears in control preparations that are not digested by a restriction endonuclease. Our procedure includes blocking nonspecific nick labeling before incubation with restriction enzymes (HpaII and HaeIII). This is achieved by incorporation of ddGTP into DNA by the Taq polymerase which lacks 3' to 5' exonuclease activity. This method gives satisfactory results because it eliminates nonspecific nick translation signals that are present after applying the methods described for animal material.