The dual origin of the nurse chamber in the ovarioles of the gall midge,Heteropeza pygmaea

Summary By following the lineage of the primordial germ cell and the mesoderm cells during the ontogeny of the gall midge,Heteropeza pygmaea, it has been determined that the nurse chamber of the polytrophic ovarioles of this insect is derived from the descendants of these cells. Such a dual origin for the nurse chamber of an insect ovary is the first of its kind to be reported.     

Karyotype and chromosomal banding pattern in Heteropeza pygmaea

The chromosomes of somatic and germ line cells of female embryos produced by paedogenesis were studied. The haploid set in somatic cells consists of one long submetacentric chromosome, one large acrocentric, one medium metacentric and two small acrocentrics. The length vs arm index karyogram makes it possible to distinguish all but the two pairs of small acrocentric chromosomes. — Attempts were made to develope a method for banding pattern visualization. The best result was obtained using trypsin which induced banding in the chromosomes of the somatic cells and occasionally also of the germ line cells. The resulting banding patterns were frequently not identical in members of a chromosome pair. There was also a variation between metaphases within an embryo as well as from different embryos. Some tentative explanations for these results are discussed.     

The chromosomal distribution of human satellite III DNA during meiosis.

In human meiotic cells DNA satellite II is located in the same sites as in mitotic cells, but in prophase the areas are less condensed. This differs from the situation in Plethodon where the sites of heavy satellite DNA are condensed throughout meiotic prophase (MacGregor and Kezer, 1971). The difference may be ascribed to the fact that Plethodon heavy satellite is pericentrically located, whereas few if any of the human satellite sites are actually at centromeres. A second difference is found in sperm, where the Plethodon satellite is located at a single site in the rear of the nucleus, while the satellite regions in mad do not have a common or constant orientation, suggesting that the respective satellites may well be functionally different.     

The organisation,nucleotide sequence,and chromosomal distribution of a satellite DNA from Allium cepa

We have investigated the organisation, nucleotide sequence, and chromosomal distribution of a tandemly repeated, satellite DNA from Allium cepa (Liliaceae). The satellite, which constitutes about 4% of the A. cepa genome, may be resolved from main-band DNA in antibiotic-CsCl density gradients, and has a repeat length of about 375 base pairs (bp). A cloned member of the repeat family hybridises exclusively to chromosome telomeres and has a non-random distribution in interphase nuclei. We present the nucleotide sequences of three repeats, which differ at a large number of positions. In addition to arrays made up of 375-bp repeats, homologous sequences are found in units with a greater repeat length. This divergence between repeats reflects the heterogeneity of the satellite determined using other criteria. Possible constraints on the interchromosomal exchange of repeated sequences are discussed.     

Early embryonic development of the dipteran insect Heteropeza pygmaea in the presence of cytoskeleton-affecting drugs

Summary Embryos of the paedogenetically reproducing gall midge Heteropeza pygmaea develop floating in the haemocoel of a so-called mother larva. The egg membranes remain permeable and the embryos increase in size during embryonic development by taking up nutrients from the haemolymph. Such embryos can be cultured in vitro, i.e. in haemolymph drops obtained from mother larvae. We tested the effects of several drugs known to interact with cytoskeletal elements on different stages of embryonic development, including cleavage and gastrulation. The drugs were added to the in vitro cultures and the effects were studied with time-lapse cine-micrography. Colchicine and vinblastine blocked cleaving eggs in metaphase stage and arrested yolk globule oscillation. In spite of such a block blastoderms once formed continued development through germ band formation and extension and also increased in size. Cytochalasin B did not affect the stage of cleavage; however, it inhibited gastrulation and subsequent morphogenetic processes and also prevented size increase. We conclude that (1) the functioning of microtubules is needed for yolk globule oscillation during cleavage interphases but not for the gastrulation processes subsequent to blastoderm formation and (2) microfilaments do not play an important role in cleavage, at least not for the orderly succession of the cleavage divisions, but are essential for the morphogenetic movements associated with gastrulation. We suggest that during cleavage a limited stock of microtubules and their precursors is responsible for both transport of chromosomes during mitoses and translocation of organelles during interphase. Yolk oscillation seems to be a secondary effect and of minor or no importance for the normal course of embryonic development.Dedicated to Professor Gerhard Krause on the occasion of his 80th birthday     

The chromosomal localisation of human satellite DNA I

The major concentrations of human satellite DNA I (1.688 g/ml) have been localised on human chromosome preparations by the technique of in situ hybridisation using radioactive complementary RNA synthesised in vitro. Chromosomes were identified by prior study using quinacrine fluorescence microscopy. The satellite DNA is concentrated, mainly in centromeric constitutive heterochromatin, on many chromosomes but is especially obvious in the fluorescent distal segment of the Y chromosome.     

The chromosomal location of human satellite DNA III

In situ hybridisation of radioactive complementary RNA has been used to localise the chromosomal distribution of human satellite DNA III. This DNA is found to be concentrated in paracentromeric heterochromatin mainly on chromosome 9 and in minor concentrations on chromosomes chiefly of the D and G groups.     

Male germ line,spermatogenesis and karyotypes of Heteropeza pygmaea winnertz (Diptera: Cecidomyiidae)

The germ line during the development of the male of the gall midge Heteropeza pygmaea was followed cell generation by cell generation. The development of the male egg begins with two meiotic divisions, followed by fusion of one of the resulting nuclei with (usually) two somatic nuclei (regulation), after which the regulated nucleus passes through 9–10 mitoses, and finally a further two meiotic divisions producing the spermatids. The chromosome numbers (determined by colchicine and air-drying techniques) of the race studied are 55 in the female germ line, very variable with a mean near 47–48 in the male germ line after regulation, 5 or 6 in the sperms, 10 in the female somatic nuclei and 5 in the male somatic nuclei. Statistical techniques for analysis of the different karyotypes are developed and a model explaining the known cytological events in Heteropeza is presented.     

Migration and division of cleavage nuclei in the gall midge,Wachtliella persicariae

Summary In the eggs ofWachtliella persicariae the cleavage nuclei move relative to the surrounding ooplasm. This active migration is caused by an organelle whose ultrastructure was studied throughout the mitotic cycle. It consists of a greatly enlarged polar cytaster derived from the mitotic apparatus, linked to the nucleus by 100 Å filaments. The microtubules of the cytaster were found only during periods of active nuclear migration, i.e., from the onset of anaphase to the early prophase of the next mitotic cycle. They are always solitary and follow the course of the astral rays, which are known to temporarily adhere to peripheral structures of the egg cell and to exert tractive forces. In contrast to the cytaster microtubules, the microtubules in the spindle are bundled and persist from early metaphase through late telophase.During ontogenesis the first migration cytaster is built up between 3 and 12 min after oviposition near the anterior egg pole, in the vicinity of the sperm nucleus. In non-inseminated eggs time lapse films show a migration cytaster to develop autonomously in a region free from nuclei, but it does not follow the normal path of the male pronucleus. In several cases the female pronucleus, which remains without a cytaster of its own, was observed to move to the cytaster generated in the absence of the male pronucleus. Whether or not it is adhering to a nucleus, the cytaster divides into two at the correct time, i.e, corresponding to the first cleavage division in fertilized eggs. In some non-inseminated eggs this type of pseudocleavage has been observed to occur repeatedly, giving rise to an increasing number of anucleate cytasters.     

Migration and division of cleavage nuclei in the gall midge,Wachtliella persicariae

Summary The control of nuclear division and migration was studied in time-lapse films of the multinucleate egg cell of a gall midge by experimental alterations of the mitotic pattern. During each cleavage cycle, a wave of randomly oriented saltations of yolk particles (WROS) is seen to travel through the ooplasm. This wave proved to be an indispensable prerequisite for the accompanying anaphase wave and for the activation of the nuclear migration cytasters: WROS cycles can occur autonomously without cleavage nuclei being present, but there is no anaphase without a WROS passing the dividing nucleus. WROSs and mitotic waves can be inverted, and the WROS cycles and the cleavage cycles can be desynchronized by temperature grandients or by locally impaired gas exchange. If a nucleus is not ready for anaphase when met by a WROS, it will only divide in the course of the next WROS. WROSs thus indicate autonomous anaphase-triggering waves governing the cleavage divisions. Rhythmic ooplasmic movements continue even if the WROSs as well as the nuclear divisions are inhibited by colchinine. The characteristics of the WROSs support the hypothesis that each of them is the visible effect of a wave of calcium release (similar to that established in vertebrate eggs) which acts locally on the microtubular system and may continue even if the WROSs are suppressed. The correlations between a possible calcium release, WROS activity, microtubule disassembly and nuclear cycle are discussed.     

Characterization and chromosomal distribution of satellite DNA sequences of the water buffalo (Bubalus bubalis).

Satellite DNA sequences were isolated from the water buffalo (Bubalus bubalis) after digestion with two restriction endonucleases, BamHI and StuI. These satellite DNAs of the water buffalo were classified into two types by sequence analysis: one had an approximately 1,400 bp tandem repeat unit with 79% similarity to the bovine satellite I DNA; the other had an approximately 700 bp tandem repeat unit with 81% similarity to the bovine satellite II DNA. The chromosomal distribution of the satellite DNAs were examined in the river-type and the swamp-type buffaloes with direct R-banding fluorescence in situ hybridization. Both the buffalo satellite DNAs were localized to the centromeric regions of all chromosomes in the two types of buffaloes. The hybridization signals with the buffalo satellite I DNA on the acrocentric autosomes and X chromosome were much stronger than that on the biarmed autosomes and Y chromosome, which corresponded to the distribution of C-band-positive centromeric heterochromatin. This centromere-specific satellite DNA also existed in the interstitial region of the long arm of chromosome 1 of the swamp-type buffalo, which was the junction of the telomere-centromere tandem fusion that divided the karyotype in the two types of buffaloes. The intensity of the hybridization signals with buffalo satellite II DNA was almost the same over all the chromosomes, including the Y chromosome, and no additional hybridization signal was found in noncentromeric sites.     

The chromosomal localisation of satellite DNA in Ptyas mucosus (Ophidia,Colubridae)

Ptyas mucosus male DNA has a repetitious DNA satellite (p= 1.700 g cm^?3) constituting 5% of the haploid genome. In situ hybridisation of radioactive complementary RNA (cRNA) has revealed that satellite sequences are located in the centromeric region of one pair of macrochromosomes and in the terminal region of 8 pairs of microchromosomes. These regions are constitutively heterochromatic as revealed by C-banding. The possibility of involvement of satellite rich microchromosomes in nucleolus organisation is discussed.     

The chromosomal distribution of the major and minor satellite is not conserved in the genusMus

The cytological distribution of the major and minor satellite first identified inMus musculus was studied in the karyotypes of three related subspecies and two other species of the genusMus. Both the major and minor satellite showed species dependent hybridization patterns. The major satellite is confined to the centromere region inM. musculus and related subspecies. However, inM spretus andM. caroli, the chromosomal arm regions contain this sequence class. In contrast the minor satellite is found at the kinetochore region inM. musculus and related subspecies but is distributed throughout the entire centromeric domain inM. spretus and appears to be excluded from the chromosomes ofM. caroli. There is an apparent correlation between the chromosomal location of these satellites and their phylogenetic relationship. Determination of the biological roles of the major and minor satellites fromM. musculus must take into account their differential chromosomal distribution in otherMus species.     

稻瘿蚊种群DNA指纹及水稻抗稻瘿蚊分子标记育种技术研究

文章综述经10年(1996~2005)研究建立的亚洲稻瘿蚊Orseolia oryzae Wood-Mason种群DNA指纹检测技术和水稻抗稻瘿蚊分子育种技术的研究成果。其核心技术包括5点:(1)在国际和国内首先用AFLP方法研究亚洲5国(中国,印度,斯里兰卡,尼泊尔和老挝)及广东省7个地点亚洲稻瘿蚊种群DNA指纹及其生物多样性;(2)分别用RAPD和微卫星SSR技术对来源于我国的水稻资源大秋其的抗亚洲稻瘿蚊基因Gm6进行精细定位;(3)分别用与Gm6基因紧密连锁的STS分子标记和SSR标记,建立了分子标记辅助选择(MAS)方法选育抗稻瘿蚊新品种的技术体系;(4)应用建立的分子标记辅助选育新技术选育出一批抗亚洲稻瘿蚊中国种群的新品系,创造出一批新种质,包括抗蚊18号、抗蚊软占等达国优3级的抗稻瘿蚊新品系6个;(5)将Gm6基因导入杂交稻恢复系,选育出4个抗稻瘿蚊两系杂交稻新组合培矮64S/KG18,培矮64S/KI41,培矮64S/AK7,培矮64S/03W16(国优2级)和1个三系杂交稻新组合抗蚊博优。其中江西省宁都市名林水稻研究所应用本研究鉴定出的带有Gm6基因的品系抗蚊青占作为抗性亲本,培育出的2个抗稻瘿蚊的两系杂交稻组合安两优青占和培两优抗占通过省级品种审定,率先选育出抗稻瘿蚊种群的杂交稻。该项研究成果为控制华南稻瘿蚊的灾害发生提供新的技术措施,选育的抗稻瘿蚊品种和新种质,在华南广为应用,取得显著的经济效益。     

Characterization and chromosomal organization of satellite DNA sequences in Picea abies

Three clones containing satellite DNA sequences were selected from a randomly sheared genomic DNA library of Picea abies (clones PAF1, PAG004P22F (2F), and PAG004E03C (3C)). PAF1 contained 7 repeats that were 37-55 bp in length and had 68.9%-91.9% nucleotide sequence similarity. Two 2F repeats were 305-306 bp in length and had 83% sequence similarity. Two 3C repeats were 193-226 bp in length and had a sequence similarity of 78.6%. The copy number per 1C DNA of PAF1, 2F, and 3C repeats was 2.7 x 106, 2.9 x 105, and 2.9 x 104, respectively. In situ hybridization showed centromeric localization of these sequences in two chromosome pairs with PAF1, all pairs but one with 2F, and three pairs with 3C. Moreover, PAF1 sequences hybridized at secondary constrictions in six pairs, while 2F-related sequences were found at these chromosome regions only in four pairs. These hybridization patterns allow all chromosome pairs to be distinguished. PAF1-related repeats were contained in the intergenic spacer (IGS) of ribosomal cistrons in all six nucleolar organizers of the complement, while sequences related to 2F were found on only one side of the rDNA arrays in four pairs, showing structural diversity between rDNA regions of different chromosomes.     

Parental origin of germ-line and somatic mutations in the retinoblastoma gene

Segregation analysis of polymorphic sites within the retinoblastoma (RB) gene and on chromosome 13, as well as the parental origin of the lost allele in the tumor, were analyzed in 24 families with RB patients. Four mutant alleles transmitted through the germ-line and seven de novo germ-line mutant alleles were identified in 11 patients with hereditary RB. Segregation analysis within the RB gene and on chromosome 13 was useful for DNA diagnosis of susceptibility to RB in relatives of hereditary patients, even if mutations were not identified. All seven de novo germ-line mutant alleles were paternally derived. The bias toward the paternal allele for de novo germ-line mutations of the RB gene was statistically significant. Seven paternal alleles and six maternal alleles were lost in 13 non-hereditary RB tumors with no bias in the parental origin of the somatic allele loss. These results suggest that the physical environment or a deficiency in DNA repair during spermatogenesis may be associated with significant risk factors for de novo germ-line mutations.