A novel approach to the analysis of the initiation of embryo development in gramineae

An in vivo model system to study the initiation of embryo development is presented. From the so-called Salmon system of wheat (alloplasmic lines with a 1BL-1RS chromosome translocation), three completely isogenic and homozygous lines were produced by selection for uniformity in about 20 selfing/backcross generations as well as between sublines of doubled haploids. The line (aestivum)-Salmon is male fertile and sexual. The lines (caudata)-Salmon and (kotschyi)-Salmon are male sterile and have a parthenogenetic capacity of about 90%. The expression of nuclear-cytoplasmic male sterility is different for the two parthenogenetic lines. The initiation of autonomous embryo development at defined developmental stages of the ovaries and the maximum degree of parthenogenesis are identical in both parthenogenetic lines as proved by the auxin test and progeny analyses. The protein patterns from ovary extracts of the three isogenic lines were identical for more than 200 spots of 2-D polyacrylamide gels, confirming their homogeneity. However, one protein (P 115.1) was found 3 days before and during anthesis only in ovaries of the parthenogenetic lines. It seems to be involved in the initiation of parthenogenesis.     

Speciation in the Artemia genus: Mitochondrial DNA analysis of bisexual and parthenogenetic brine shrimps

From the cloned mitochondrial DNAs (mtDNAs) isolated from two bisexual species, one Mediterranean, Artemia salina, and one American, Artemia franciscana, and two parthenogenetic (diploid and tetraploid) strains of Artemia parthenogenetica collected in Spain, physical maps have been constructed and compared. They are extremely different among themselves, much more than the differences between Drosophila melanogaster and D. yakuba and in the same range of different mammalian species such as mouse/rat or man/cow. The nucleotide sequences of two regions of mtDNA encoding parts of the cytochrome c oxidase subunit I (COI) and cytochrome b (Cytb) genes have been determined in the two bisexual species and the two parthenogenetic strains. Comparisons of these sequences have revealed a high degree of divergence at the nucleotide level, averaging more than 15%, in agreement with the differences found in the physical maps. The majority of the nucleotide changes are silent and there is a strong bias toward transitions, with the CT substitutions being highly predominant. The evolutionary distance between the two Artemia parthenogenetica is high and there is no clear relationship with any of the bisexual species, including the one present nowadays in Spain. Using a combination of molecular (mtDNA) and morphological markers it is possible to conclude that all of these Artemia isolates should be actually considered as belonging to different species, even the two Artemia parthenogenetica diploidica and tetraploidica.On sabbatical leave from Departamento de Bioquímica, Facultad de Veterinaria, Universidad Complutense de Madridearly Italian artemiologists to designate the Medi-Beatriz Batuecas died in an accident during the Christmas holy days of 1988 after she had initiated this workCorrespondence to: R. Garesse     

Enigma of apogamety

Summary Apogamety, the occurrence of which has been either denied or criticized by some authors, presents several problems that should be carefully considered in argumentation. Gynogenesis, which comprises zygotic, parthenogenetic, and apogametic embryos, an actively developing branch of tissue culture, is of no help here being itself subject to similar criticism. The discussion on apogamety awaits methodological progress which would yield answers to the following questions: are all cells of embryo sac potentially gametic; in what way are the mechanisms of cell differentiation and specialization of cells within embryo sac correlated with their totipotency; what are the limitations of plant cell totipotency in embryo sacs. The above problems of apogamety are not only an enigma for embryology and reproduction of angiosperms but they belong also to the crucial problems of the general biology which might be solved by studies of corresponding molecular mechanisms and experiments in tissue culture.     

An exotic species spreads through Europe: Tygarrup javanicus (Chilopoda: Geophilomorpha: Mecistocephalidae) is reported from the Slovakia and the Czech Republic

Geophilomorph centipedes, due to their soil life style, are frequently transported with plants. This report tries to underline some adaptations that facilitate them being introduced and becoming alien species. Tygarrup javanicus (Attems, 1907), tropical species reported recently from different greenhouses in Central Europe, is used as an example of such a spreading species.     

绵羊胞内单精子注射技术

本文的主要目标是建立绵羊胞内单精子注射技术(intracyioplasmic sperm injection,ICSI)。并且尝试了ICSI技术生产绵羊转基因胚胎的可能性。实验1比较了绵羊卵母细胞孤雌激活的3种化学方法。结果显示,Ionomvcin/3 h/6-dimethylaminopurine(6-DMAP)和Ionomycin激活胚的卵裂率(71.7%和91.8%)和囊胚率(17.4%和11%)显著(P<0.01)高于Ca~(2+)激活胚(18.4%和0)。Ionomycin/3 h/6-DMAP激活胚的囊胚发育形态和比例相对较高。实验2中,活精子注射至卵母细胞质后,用Ionomycin/3 h/6-DMAP激活,排出第二极体(PB2)的71枚ICSI胚胎用SOFaaBSA溶液培养,卵裂率为71.8%(51/71),显著(P<0.01)高于体外受精(41.4%,IVF)和阴性对照胚胎(30.2%,sham-ICSI)。培养7天后,sham-ICSI组没有囊胚生成;ICSI和IVF胚胎的囊胚率分别为7.0%(5/71)和16.1%(9/56),两者差异不显著(P>0.05)。这些ICSI囊胚在冷冻前后均能孵化,显示初步建立了绵羊ICSI技术。另外,我们探索了ICSI技术生产转基因胚胎的可能性,-20℃冻融1次的死精子与pEGFP-N1质粒共注射,33枚2-细胞期ICSI胚胎中,2枚可见GFP蛋白。其中1枚停止发育,另外1枚继续发育至16-细胞期,仍然可见GFP基因的表达。4枚解冻的ICSI囊胚手术移植给2只发情同期化受体绵羊,60天时,B超未见怀孕。本文初步结果表明,ICSI技术生产转基因绵羊有可能性,需深入研究。     

First evidence of polyploidy in Psylloidea (Homoptera,Sternorrhyncha): a parthenogenetic population of <Emphasis Type="Italic">Cacopsylla myrtilli</Emphasis> (W. Wagner, 1947) from northeast Finland is apomictic and triploid

This paper reports results of the first cytogenetic study of parthenogenetic psyllids, carried out on an asexual population of the holarctic species Cacopsylla myrtilli W. Wagner from northeast Finland. Preparations of mature eggs extracted from females revealed 39 univalent chromosomes in prophase and metaphase cells. Hence, female meiosis is of apomictic type and replaced by a modified mitosis. The karyotype consists of 3n = 39 (36 + XXX). Clearly, the population is triploid, the haploid number being n = 12 + X as characteristic of the genus Cacopsylla as a whole. As typical for Psylloidea, the chromosomes are holokinetic, only slightly varying in size and without any visible markers, rendering impossible the precise identification of triplets of homologous chromosomes in the triploid complement. The distribution of bisexual and parthenogenetic populations of C. myrtilli throughout the world is briefly given, and a possible origin of the triploid parthenogenetic population is discussed.     

Parthenogenesis in non-rodent species: developmental competence and differentiation plasticity

An oocyte can activate its developmental process without the intervention of the male counterpart. This form of reproduction, known as parthenogenesis, occurs spontaneously in a variety of lower organisms, but not in mammals. However, it must be noted that mammalian oocytes can be activated in vitro, mimicking the intracellular calcium wave induced by the spermatozoon at fertilization, which triggers cleavage divisions and embryonic development. The resultant parthenotes are not capable of developing to term and arrest their growth at different stages, depending on the species. It is believed that this arrest is due to genomic imprinting, which causes the repression of genes normally expressed by the paternal allele. Human parthenogenetic embryos have recently been proposed as an alternative, less controversial source of embryonic stem cell lines, based on their inherent inability to form a new individual. However many aspects related to the biology of parthenogenetic embryos and parthenogenetically derived cell lines still need to be elucidated. Limited information is available in particular on the consequences of the lack of centrioles and on the parthenote's ability to assemble a new embryonic centrosome in the absence of the sperm centriole. Indeed, in lower species, successful parthenogenesis largely depends upon the oocyte's ability to regenerate complete and functional centrosomes in the absence of the material supplied by a male gamete, while the control of this event appears to be less stringent in mammalian cells. In an attempt to better elucidate some of these aspects, parthenogenetic cell lines, recently derived in our laboratory, have been characterized for their pluripotency. In vitro and in vivo differentiation plasticity have been assessed, demonstrating the ability of these cells to differentiate into cell types derived from the three germ layers. These results confirmed common features between uni- and bi-parental embryonic stem cells. However data obtained with parthenogenetic cells indicate the presence of an intrinsic deregulation of the mechanisms controlling proliferation vs. differentiation and suggest their uni-parental origin as a possible cause.     

Genetically identical copies of the mulberry silkworm

Summary Genetically identical copies of the silkworm female obtained via ameiotic parthenogenesis are insignificantly depressed by artificial reproduction, and notwithstanding their isogenous pattern, they show the same variability in quantitative characters as in the heterogenous forms.By means of androgenesis, genetically identical copies of strictly homozygous males were obtained; the latter were produced via meiotic parthenogenesis. The androgenetic clones of these males were transformed by backcrosses into homozygous bisexual lines. Viability and cocoon weight in these lines were higher than those in androgenetic clones of similar genotype, but they were far from the norm due to the depressing effect of semilethals left in them in a homozygous state. An extremely low phenotypical variability of quantitative characters is observed in the isogenous hybrids F₁ obtained from crossing two genetically dissimilar strictly homozygous individuals. These hybrids make excellent material for phenogenetical studies. The female and male isogenous clones have an increased combining ability which is acquired in the course of their selection for high disposition towards complete parthenogenesis. For practical purposes it is proposed to take a female and a male which when mated produce a high-quality family and clone them separately, the first via parthenogenesis, the latter via androgenesis. After mating these reproduced bisexual clones, it will be possible to obtain in successive generations a vast number of families repeating the prominent productivity of the initial family.     

Effectiveness of slow freezing and vitrification for long-term preservation of mouse ovarian tissue

This study was conducted to evaluate the interaction between cryo-damage and ART outcome after cryopreservation of mouse ovarian tissues with different methods. Either a vitrification or a slow freezing was employed for the cryopreservation of B6CBAF1 mouse ovaries and follicle growth and the preimplantation development of intrafollicular oocytes following parthenogenesis or IVF were monitored. Both cryopreservation protocols caused significant damage to follicle components, including vacuole formation and mitochondrial deformities. Regardless of the cryopreservation protocols employed, a sharp (P < 0.0001) decrease in follicle viability and post-thaw growth was detected. When IVF program was employed, significant (P < 0.05) decrease in cleavage and blastocyst formation was notable in both modes of cryopreservation. However, such retardation was not found when oocytes were parthenogenetically activated. In the IVF oocytes, slow freezing led to better development than vitrification. In conclusion, a close relationship between cryopreservation and ART methods should be considered for the selection of cryopreservation program.     

Variability in life history traits of the aphid,Acyrthosiphon pisum (Harris), from sexual and asexual populations

Many aphid species have shown remarkable adaptability by invading new habitats and agricultural crops, although they are parthenogenetic and might be expected to show limited genetic variation. To determine if the mode of reproduction limits the level of genetic variation in adaptively important traits, we assess variation in 15 life history traits of the pea aphid, Acyrhosiphon pisum (Harris), for five populations sampled along a north-south transect in central North America, and for three traits for three populations from eastern Australia. The traits are developmental times and rates as affected by temperature, body weights as affected by temperature, fecundity, measures of migratory tendency, and photoperiodic responses. The most southerly population from North America is shown to be obligately parthenogenetic, as are the Australian populations, and the four more northerly North American populations are facultatively parthenogenetic with the number of parthenogenetic generations per year increasing from north to south. The broad-sense heritabilities of life history traits varied from 0.36 to 0.71 for nine quantitive traits based on a comparison of within-and between-lineage variances. Using these traits, 7–13 distinct genotypes (i.e. clones) were identified among each of the 18 lines sampled from the North American populations, but the number did not differ significantly among populations. The level of genetic variation differed from trait to trait. For 4 of 12 quantitative traits, the level of variation in the obligately parthenogenetic population from North America was lowest, but significantly lower than all the sexual populations for only 1 trait. The obligately parthenogenetic population had the highest level of genetic variation for two traits, and had intermediate levels for the others. The most northerly population, which was sexual and had relatively few parthenogenetic generations each year, had the lowest level of variation for 5 of 12 traits and the highest level of variation for 2 traits. There was no decline in variability from north to south correlated with the increase in the annual number of parthenogenetic generations. The Australian populations showed no less variation than the North American populations for two of three traits, although the pea aphid was introduced to Australia only 5 years prior to the study, whereas the aphid has been in North America for at least 100 years. The mode of reproduction has not had a substantial impact on the level of genetic variation in life history traits of the pea aphid, but there are population-specific factors that effect the level of variation in certain traits.