Development and morphology of teratocytes in Encarsia berlesei and Encarsia citrina: first record for Chalcidoidea

In several species of hymenopteran parasitoids of the superfamilies of Ichneumonoidea and Platygastroidea, the membrane enveloping the parasitoid embryo dissociates at hatching into a number of cells, called teratocytes, which autonomously develop in the host haemolymph. In this work we report for Encarsia berlesei and Encarsia citrina (Hymenoptera: Chalcidoidea), the dissociation of the extraembryonic membrane into cells whose morphological and embryological features correspond to those of teratocytes. In E. berlesei the membrane dissociated at hatching into 4-9 larger cells (100 microm diameter) and about 10 smaller cells (60 microm), which scarcely doubled their size during maturation. In E. citrina the membrane dissociated into five large cells (250 microm) which did not grow appreciably. Ultrastructural investigation of the dissociated cells in E. berlesei revealed that their surface was covered by microvilli, whose density and length increased from the egg stage to the 12 h following hatching. During the same period, rough endoplasmic reticulum evolved from a parallel profile to that of the cisternal type, while abundant vesicles represented the dominant cytological feature. The ploidy level of these cells ranged between 8c and 140c at hatching, but increased to 40c-350c at maturation. These findings provide the first clear evidences for the presence of teratocytes in the superfamily Chalcidoidea.     

三个鲫品系DNA含量的比较研究

采用流式细胞术 (FCM)对红鲫、彭泽鲫、异育银鲫进行红血球DNA含量的检测分析比较 ,以鉴定它们的倍性。结果显示 ,红鲫红血球的DNA含量是 3 0pg ,彭泽鲫是 4 7pg ,异育银鲫是 4 8pg。显而易见 ,彭泽鲫的DNA含量是二倍体红鲫的 1 57倍 ,异育银鲫的DNA含量是红鲫的 1 6倍。采用肾细胞直接制作染色体的方法进行红鲫、彭泽鲫、异育银鲫的染色体倍性鉴定 ,结果红鲫的染色体数目是 10 0 ,为二倍体 (2n =10 0 ) ,彭泽鲫的染色体数目是 162 ,为三倍体 (3n =162 ) ,异育银鲫的染色体数目是 156— 162 ,为三倍体 (3n =156— 162 )。研究证明 :不同品系鲫的DNA含量高低与染色体的倍性有显著的正相关性     

Influence of ploidy on plastome mutagenesis in Nicotiana

Summary A clear influence of ploidy was observed on the frequency of both spontaneous and nitroso-methylurea (NMU) induced, streptomycin-resistant, adventitious shoots developing on leaf explants of Nicotiana tabacum and N. plumbaginifolia. At nearly all NMU levels employed a significantly higher yield of resistant shoots was obtained from haploid compared with diploid leaf strips. At 1 mM NMU the differences were not significant and were absent when a high (1000 mg/1) selective concentration of streptomycin sulphate was used. The influence of ploidy is discussed in relation to the possible effect of plastome copy number on mutagenesis and sorting out of resistant plastids.     

Abundance of microtubules in preprophase bands of some Triticum species

Root tip procambial cells of Triticum speltoides, T. tauschii, T. turgidum and T. aestivum have been investigated ultrastructurally for the detection of preprophase microtubule bands (PMBs) and to estimate the number of microtubules comprising the bands. The species selected are phylogenetically related but differ in the ploidy level. It was found that all species develop well-defined PMBs prior to mitosis. Estimations of microtubule abundance in the PMBs was carried out in midpreprophase cells, a stage judged by a feature of the nucleus in which electron-transparent canals are formed around the initial condensations of the chromatin material and the nucleoli. Triticum speltoides bears the smaller average number of microtubules per PMB and T. aestivum the greater. The results indicate that the increase follows the upgrade of the number of chromosome sets. It is suggested that the average number of microtubules of PMBs is related to the ploidy level.Abbreviation PMB preprophase microtubule band     

柑橘体细胞杂种有性后代的创造及杂种鉴定

以异源四倍体柑橘体细胞杂种[哈姆林甜橙(Citrus sinensis Osbeck) 粗柠檬(C.jambhiri Luss)]为父本与二倍体单胚类型宜本杂4号[华农本地早橘(C.reticulata Blanco)×宜昌橙(C.ichangen- sis Swingle)]杂交,采用胚抢救技术获得了110株有性后代植株,通过染色体计数及倍性分析仪鉴定,其中93株植株为3倍体,余下的17林植株为2倍体。RAPD分析表明:获得的有性后代植株均为杂种。     

Cytogenetics of protoplast cultures of Brachycome dichromosomatica and Crepis capillaris and regeneration of plants

Summary Callus derived protoplasts of Brachycome dichromosomatica (2n=2x=4) and Crepis capillaris (2n=2x=6) have been regenerated into karyologically normal plants, i.e. plants without visible alterations of the diploid chromosome set. However, metaphase analysis of protoplast cultures derived from both callus as well as mesophyll cells showed karyological changes in the overwhelming majority of cells in both species leading to multinucleated, polyploid and aneuploid cells. Furthermore, callus derived protoplasts sometimes exhibited changes at the chromosome level as indicated by translocations. The vast majority of aberrant karyotypes arose from failures during mitosis and cytokinesis, pointing to inadequate microtubules as a possible underlying cause. Karyological events of the kind described herein greatly affect the plating efficiency of isolated protoplasts and the viability of protoplast derived calli. Plant regeneration, although demonstrated in this study for the first time in both species, seems to be limited to rarely occurring, protoplast-derived colonies with a relatively stable genome. Our experiments, performed with chromosomal model species, emphasize the need for controlled, non-mutagenic culture conditions.     

辣椒游离小孢子细胞团培养的胚状体形成

从预培养15天后的花药中机械游离小孢子及其细胞团,经28℃液体悬浮暗培养．30天后,获得了自球形期胚到子叶期胚发育程度不等的各类胚状体。从12个花药中可以形成高达22个胚状体,且子叶期胚的比例约为23％。显微镜检表明,这些胚状体来自游离的小孢子细胞．经核的对称分裂形成多核细胞或者早期形成多细胞团,最后经细胞的分裂分化形成。胚状体体表具毛,活力有差异。在适当培养基上,具活力的鱼雷期及子叶期胚状体均能发育成正常植株。7℃、32℃、35℃8天的胁迫处理均能诱导小孢子胚状体发生。但花药培养中7℃、35℃处理下的出胚率较32℃下高,而游离小孢子细胞团培养中以35℃、32℃下较好。7℃处理下获得的胚状体数很少．对产生这种现象的原因进行了探讨。出胚率在基因型间,不同胁迫处理温度间表现明显差异。而在温度处理的不同天数间差异不明显。流式细胞仪对再生株真叶的DNA含量分析表明．获得的再生株中具有单倍体、双单倍体以及单倍一双倍嵌合体植株。本结果为进一步开展辣椒雄性生殖途径的胚状体发育研究。提高辣椒成熟胚状体的频率提供了实验体系。     

Embryonic stem cell/fibroblast hybrid cells with near-tetraploid karyotype provide high yield of chimeras

Ten primary clones of hybrid cells were produced by the fusion of diploid embryonic stem (ES) cells, viz., line E14Tg2aSc4TP6.3 marked by green fluorescent protein (GFP), with diploid embryonic or adult fibroblasts derived from DD/c mice. All the hybrid clones had many characteristics similar to those of ES cells and were positive for GFP. Five hybrid clones having ploidy close to tetraploidy (over 80% of cells had 76–80 chromosomes) were chosen for the generation of chimeras via injection into C57BL blastocysts. These hybrid clones also contained microsatellites marking all ES cell and fibroblast chromosomes judging from microsatellite analysis. Twenty chimeric embryos at 11–13 days post-conception were obtained after injection of hybrid cells derived from two of three clones. Many embryos showed a high content of GFP-positive descendents of the tested hybrid cells. Twenty one adult chimeras were generated by the injection of hybrid cells derived from three clones. The contribution of GFP-labeled hybrid cells was significant and comparable with that of diploid E14Tg2aSc4TP6.3 cells. Cytogenetic and microsatellite analyses of cell cultures derived from chimeric embryos or adults indicated that the initial karyotype of the tested hybrid cells remained stable during the development of the chimeras, i.e., the hybrid cells were mainly responsible for the generation of the chimeras. Thus, ES cell/fibroblast hybrid cells with near-tetraploid karyotype are able to generate chimeras at a high rate, and many adult chimeras contain a high percentage of descendants of the hybrid cells. A. A. Kruglova and E. A. Kizilova contributed equally to this work. This study was financially supported by grants from the Russian Academy of Sciences, Siberian Branch 5.2 and 14.0.     

Genetic drift on networks: ploidy and the time to fixation

Genetic drift in finite populations ultimately leads to the loss of genetic variation. This paper examines the rate of neutral gene loss for a range of population structures defined by a graph. We show that, where individuals reside at fixed points on an undirected graph with equal degree nodes, the mean time to loss differs from the panmictic value by a positive additive term that depends on the number of individuals (not genes) in the population. The effect of these spatial structures is to slow the time to fixation by an amount that depends on the way individuals are distributed, rather than changing the apparent number of genes available to be sampled. This relationship breaks down, however, for a broad class of spatial structures such as random, small-world and scale-free networks. For the latter structures there is a counter-intuitive acceleration of fixation proportional to the level of ploidy.