A newly identified chemotactic sexual pheromone from Closterium ehrenbergii

 In sexual reproduction of Closterium ehrenbergii, pairing with the sexual partner cells is the first process observed. A cell migration-inducing activity, specific for mating-type plus (mt⁺; NIES-228) cells, was detected in the culture medium of mating-type minus (mt^–; NIES-229) cells. Light was necessary for production of the active substance by mt^– cells and for migration of mt⁺ cells. The active substance was heat-labile and had an apparent molecular mass of 20 kDa, as determined by gel filtration. A protein of 20 kDa was detected in the active fraction of gel filtration after sodium dodecyl sulfate polyacrylamide gel electrophoresis. Based on these results, it is proposed that a chemotactic sexual pheromone involved in the formation of sexual pairs of cells is secreted by mt^– cells of C. ehrenbergii and is proteinaceous, like other sexual pheromones secreted by Closterium species. Received: 31 July 1997 / Revision accepted: 25 November 1997     

Physiological characterization of the sex pheromone protoplast-release-inducing protein from the Closterium peracerosum-strigosum-littorale complex (Charophyta)

In the unicellular charophycean alga Closterium peracerosum‐strigosum‐littorale complex, the protoplast‐release‐inducing protein (PR‐IP), a sex pheromone responsible for gametic protoplast release from mating‐type minus (mt^–) cells, was found to stimulate secretion of mucilage from the cells. Induction of sexual cell division by PR‐IP was also confirmed. Bioassays were used to determine the minimum doses required to induce these functions, revealing that 5 · 10^?16 M of PR‐IP stimulated mucilage secretion, and that 5 · 10^?10 M of PR‐IP were required for protoplast release. Exposure of the cells to 5 · 10^?11 M of PR‐IP resulted in the induction of sexual cell division as well as mucilage secretion. These results strongly suggest that PR‐IP is a multifunctional pheromone that independently promotes multiple steps in conjugation at the appropriate times through different induction mechanisms.     

Dissection of conjugants and mating type plus and minus cells in selfing clones of the isogamous green alga Closterium ehrenbergii

In the sexual reproduction of the green alga Closterium ehrenbergii, two sexually competent cells that are morphologically indistinguishable from the vegetative cells first come close to each other to form a sexually interacting pair. Each then divides into two gametangial cells. Isogamous conjugation occurs between nonsister gametangial cells of the two resulting pairs. With unusual selfing clones derived from a certain cross of heterothallic strains, we dissected apart a pair of gametangial cells that had already been united together by a delicate transparent tube, into which each gametangial cell was going to develop its conjugation papilla. In spite of such a degree of differentiation, when each was cultured in fresh medium, individual gametangial cells could dedifferentiate into vegetative cells and form subclones. By crossing such subclones with standard stable heterothallic mating-type strains, we show that each selfing clone of this alga actually produces both stable mt ⁺ and stable mt ^- cells, in addition to unstable mt ^- cells with selfing potency, during its mitotic vegetative growth. Although the selfing in C. ehrenbergii studied here differs in certain points from true homothallism, the results of the present study provide insight into how homothallism might have evolved from heterothallism.     

Observation on the Sexual Reproduction of Peridinium bipes Stein (Dinophyceae)

Sexual reproduction of the dinoflagellate Peridinium bipes Stein was observed. At the late growth season (from late March to early April), the small, unarmored motile cells i.e. gametes are produced by division of the cate cell. Isogamy occurs in this species. Two gametes in fusion are morphologically indistinguishable, but their behavior are different. Before fusion, two gametes are connected by a transparent granular structure and move quickly for about ten minutes. When the plasmogamy almost completes, the fusing cells stop moving for a while and the transverse flagellum of one gamete is cast off. By staining with modified carbol fuchsin, it was proved that the karyogamy takes place soon after plasmogamy and the change of chromosomes in this period was also observed. The zygote keeps motile for about 14 days before casts off its two flagella and becomes aplanozygote. During this period i.e. planozygote stage, the zygote enlarges from 55 × 50 μm to 75 ×70 μm, intercolary bands connecting thecai plates widen, lots of oil droplets are produced as storage granules. After sinking to the bottom of flask, aplano- zygote continues changing: exospore wall is cast off, mesospore wall and endospore wall are thickened, oil droplets turn to starch grains, protoplast contracts and becomes spherical, a large red lipid granule, perhaps eye-spot, appears. Afterward, aplanozygote has become hypnozygote i.e. resting cyst. The type of sexual reproduction, the amphiesma of the zygote, the resistance of hypnozygotic wall to acid and alkali, the relationship between fossil dinoflagellate remain and the wall of hypnozygote were discussed. It was also considered that the three formas of P. bipes named by Huber-Pestalozzi were the different stages of zygote development.     

REPRODUCTIVE ISOLATION BY SEX PHEROMONES IN THE CLOSTERIUM PERACEROSUM–STRIGOSUM–LITTORALE COMPLEX (ZYGNEMATALES,CHAROPHYCEAE)1

The Closterium peracerosum–strigosum–littorale (C. psl.) complex consists of unicellular algae and is known to be composed of several reproductively isolated mating groups of heterothallic strains. Group I‐E is completely isolated from mating groups II‐A and II‐B, groups II‐A and II‐B are partially isolated from each other, and only mating‐type plus (mt⁺) cells of group II‐A and mating‐type minus (mt^?) cells of group II‐B form zygotes. Based on the alignment of 1506 group I introns, significant phylogenetic relationships were observed among mating groups II‐A and II‐B, while mating group I‐E was distant from groups II‐A and II‐B. Sexual cell division in both mating‐type cells of group II‐A was stimulated in conditioned media in which cells of group II‐B had been cultured. When mt^? cells of group II‐B were stimulated in conditioned medium derived from group II‐A, mt⁺ cells of group II‐B did not respond to the conditioned medium. Conditioned media derived from group I‐E did not exhibit sexual cell division (SCD)–inducing activity against any strain except those within its own group. From the alignment of deduced amino acid sequences from orthologous protoplast‐release‐inducing protein (PR‐IP) Inducer genes, we detected a significant similarity among groups II‐A and II‐B, and mating group I‐E had low similarity to other mating groups. The existing degree of reproductive isolation can be partially explained by differences in molecular structures and physiological activities of sex pheromones of these heterothallic mating groups.     

Mating system in the tarantula spider Eupalaestrus weijenberghi (Thorell, 1894): evidences of monandry and polygyny

The road tarantula Eupalaestrus weijenberghi shows a strongly female-biased sex ratio since adult females live several years while adult males live only for 2 months. In this scenario selective males could be expected. However, several factors such as the rates of reproduction of each sex, degree of sexual selectivity and synchronicity of female receptiveness determine the operational sexual ratio and mating system of the species. Our objective was to determine the mating rates and mating tactics for females and males of E. weijenberghi and their variation throughout the reproductive period. Four hundred sexual encounters among 20 females and 20 males in all possible pair-wise combinations were carried out during 29 days, a brief but intense experimental period, as it also occurs in the field. Mating success differed strongly between sexes. Females mated once: five females mated at the first attempt, eight initially rejected males and copulated in subsequent attempts. Half of the males did not copulate and the others copulated 1-3 times. Mated females actively rejected males. Results indicate a mating system with monogamous females and polygamous males. Not all the females were receptive in every reproductive season. We suggest that female monogamy drastically affects the operational sex ratio, since several females were unreceptive after a single copulation, directly diminishing the male potential reproductive rate. This is the first experimental approach to estimate tarantula mating systems, their determinants and the consequences of the strategies shown by each sex.     

Studies on Sexual Reproduction and Vegetative Propagation of Eleutherococcus senticosus (Araliaceae)

Eleutherococcus senticosus has two reproduction modes: vegetative propagation and sexual reproduction. In this paper, flower number per inflorescence, seed set and plump seed ratio,some aspects of rhizome morphology and morphogenesis of three morphs with different filament lengths are reported, and the breeding system is studied by hand-pollination. It is clear from the two tables presented in the present paper that plants with long filaments cannot fruit, plants with short filaments can fruit annually, but their seed setting ratio is different in different years and different habitats, plants with medium-long filaments can also fruit but their plump seed ratio may be low. In ordinary years, the seed setting ratio of plants with short filament reaches 40% and plants at the edge of forests have slightly higher seed setting ratio than those on cleared area and those in natural secondary forests. Considering the existence of only a few populations with medium-long filaments in the area studied,we conclude that most of seeds in Eleutherococcus senticosus should be the products of cross pollination;and with the coexistence of vegetative propagation and sexual reproduction, the species of Eleutherococcus senticosus should not be endangered,if not destructed by human beings.     

SEXUAL PROCESSES AND PHYLOGENETIC RELATIONSHIPS OF A HOMOTHALLIC STRAIN IN THE CLOSTERIUM PERACEROSUM–STRIGOSUM–LITTORALE COMPLEX (ZYGNEMATALES,CHAROPHYCEAE)1

Members of the Closterium peracerosum–strigosum–littorale (C. psl.) complex are unicellular charophycean algae in which there are two modes of zygospore formation, heterothallic and homothallic. A homothallic strain of Closterium (designation, kodama20) was isolated from a Japanese rice paddy field. Based on alignment of the 1506 group‐I introns, which interrupt nuclear SSU rDNAs, homothallic kodama20 is most closely related to the heterothallic mating group II‐B, which is partially sexually isolated from group II‐A. Time‐lapse photography of the conjugation process in kodama20 revealed that most of the observed zygospores originated from one vegetative cell. The sexual conjugation process consisted of five stages: (1) cell division resulting in the formation of two sister gametangial cells from one vegetative cell, (2) formation of a sexual pair between the two sister gametangial cells (or between gametangial cells of another adjoined individual), (3) formation of conjugation papillae, (4) release of gametic protoplasts from both members of a pair, and (5) formation of the zygospore by protoplast fusion. For conjugation to progress, the cell density and light condition in the culture was critical. We suggested the presence of a conjugation promotion factor.     

离体受精作为技术平台在被子植物有性生殖研究中的应用

被子植物的离体受精10a前在玉米中已获得成功,尽管目前只在玉米获得完全成功和小麦获得部分成功,但离体受精技术的研究成果非常显著。目前离体受精技术已被用于其他的研究,如用分离的精细胞和卵细胞筛选配子细胞的特异基因和蛋白质：研究合子细胞被激活的机理：用不同种植物的精、卵细胞体外融合进行新的远缘杂交尝试;利用合子细胞易分裂和胚胎发生特征探索用其作为转基因研究的受体细胞等。以离体受精技术为基础在高等植物发育生物学和生殖生物学领域的基础研究和应用探索显示了巨大潜力。介绍了离体受精技术在被子植物有性生殖的研究成果和应用前景,为研究和利用被子植物有性生殖过程中的生殖细胞特征提供线索。     

Behavioural Interactions Selecting for Symmetry and Asymmetry in Sexual Reproductive Systems of Eusocial Species

Understanding the life-history complex of eusociality has remained an enduring problem in evolutionary ecology, partially because natural selection models have considered traits in relative isolation. I aim for a more inclusive model that uses ecological interactions to predict the evolutionary existence of sexual reproduction, sexual reproduction asymmetry, and sex ratios in eusocial species. Using a two-level selection process, with within-population selection on the sex ratio of the sexual caste and between-population selection on the worker sex ratio and the degree of sexual reproduction asymmetry, it is found that a male-haploid genome and a worker caste of pure females is the evolutionary optimum of most initial conditions when, like in eusocial hymenoptera, there is no pair bond between the sexual male and female. That a diploid genome and a worker caste with both males and females is the evolutionary optimum of most initial conditions when, like in eusocial termites, there is a pair bond. That sex-linked genomes may evolve in diploid eusocials, and that the model will not generally maintain sexual reproduction by itself. These results hold for ploidy-levels that behave as quantitative or discrete traits, over a relatively wide range of the relative investment in a sexual male versus sexual female, and for partial sexual systems where the genomic portion with diploid inheritance is either fixed or random.     

Properties of cell surface carbohydrates in sexual reproduction of the Closterium peracerosum–strigosum–littorale complex (Zygnematophyceae,Charophyta)

The Closterium peracerosum–strigosum–littorale complex is a best characterized zygnematophycean green alga with respect to the process of sexual reproduction. Intercellular communication mediated by two sex pheromones has been well documented in this organism, but information concerning direct cell–cell recognition and fusion of cells involved in conjugation processes has not yet been clarified. In this study, we examined the properties of cell surface carbohydrates in vegetative and reproductive cells using a variety of fluorescein isothiocyanate labeled lectins as probes. Among 20 lectins tested, 10 bound to the Closterium cell surface, and eight of these were specific for the cells involved in sexual reproduction. In addition, some of the lectins inhibited the progress of zygote formation. In particular, Lycopersicon esculentum lectin (LEL) and ConcanavalinA (ConA) considerably inhibited zygote formation (23.6% and 0% of zygotes formed, respectively, compared with the control). LEL mainly accumulated on conjugation papillae and on the surface and lumens of empty cell walls remaining after zygote formation. ConA bound to both vegetative and sexually reproductive cells and strongly accumulated on the conjugation papillae of the latter, indicating ConA binding material(s) are non‐specifically present in Closterium cells but some of the material(s) would be essential for zygote formation. These results suggest that different carbohydrates specifically recognized by these lectins are involved in cell recognition and/or fusion during conjugation processes in the C. psl. complex.     

Active female courtship behavior and male nutritional contribution to female fecundity in Bruchidius dorsalis (Fahraeus) (Coleoptera: Bruchidae)

Mating behavior and the male's contribution to female fecundity were studied in the bean weevil Bruchidius dorsalis (Fahraeus) (Coleoptera: Bruchidae) in comparison with two other species, Callosobruchus chinensis (which infests stored beans) and Kytorhinus sharpianus (which feeds on wild legumes). Only females of B. dorsalis showed multiple mating and characteristic precopulatory behavior that appeared to solicit the male's nutritious secretion. In contrast, all females of the other two species did not copulate multiply and did not show such precopulatory behavior. In B. dorsalis, the decrement of male body weight just after copulation indicated that seminal fluid weighing as much as approximately 7% of the male's body weight was transferred to the female. Fecundity was more than eight times higher in females that had copulated ten times than in females that had copulated only once, indicating that males paid most of the nutritional cost of egg production. These facts suggest that the sex role is reversed in B. dorsalis. Received: May 22, 1998 / Accepted: July 19, 1999     

Evidence for Positive Selection on a Sexual Reproduction Gene in the Diatom Genus Thalassiosira (Bacillariophyta)

Single likelihood ancestor counting (SLAC), fixed effects likelihood (FEL), and several random effects likelihood (REL) methods were utilized to identify positively and negatively selected sites in sexually induced gene 1 (Sig1) of four different Thalassiosira species. The SLAC analysis did not find any sites affected by positive selection but suggested 13 sites influenced by negative selection. The SLAC approach may be too conservative because of low sequence divergence. The FEL and REL analyses revealed over 60 negatively selected sites and two positively selected sites that were unique to each method. The REL method may not be able to reliably identify individual sites under selection when applied to short sequences with low divergence. Instead, we proposed a new alignment-wide test for adaptive evolution based on codon models with variation in synonymous and nonsynonymous substitution rates among sites and found evidence for diversifying evolution without relying on site-by-site testing. The performance of the FEL and REL approaches was evaluated by subjecting the tests to a type I error rate simulation analysis, using the specific characteristics of the Sig1 data set. Simulation results indicated that the FEL test had reasonable Type I errors, while REL might have been too liberal, suggesting that the two positively selected sites identified by FEL (codons 94 and 174) are not likely to be false positives. The evolution of these codon sites, one of which is located in functional domain II, appears to be associated with divergence among the three major Thalassiosira lineages. Electronic Supplementary Material Electronic Supplementary material is available for this article at and accessible for authorised users. [Reviewing Editor: Dr. Martin Kreitman]     

Induction of sexual behavior in male fish (Rhodeus ocellatus ocellatus) by amino acids

Summary A new function of amino acid in fish behavior was found. Amino acids induced sexual behavior in male rose bitterling (Rhodeus ocellatus ocellatus). Two types of sexual behavior which were pecking and sperm release were observed. Amino acids are known as feeding stimulants in some fish. The pecking behavior of male fish induced by amino acids is similar to the feeding behavior but it was sexual. Only male bitterling showed pecking and sperm release but the female showed no response to the amino acids. 10 out of 20 amino acids induced sexual behavior and both pecking and sperm release were induced by the same amino acids. These two kinds of behavior changed alternately depending on the light conditions. It is of interest that non-specific material such as some amino acids function like sex pheromone.     

Natural populations of the Closterium ehrenbergii (Desmidiales, Chlorophyta) species complex in Nepal

Several natural populations of the Closterium ehrenbergii Meneghini ex Ralfs species complex were collected in Nepal, in October–December 1982. Water temperature and pH were also recorded. Clonal isolates from these populations were identified to one of four mating groups (H, I, J and M) by test crossing with standard mating-type strains of known mating groups. Groups H and M have smooth walled zygospores, while Groups I and J have scrobiculated zygospore walls. Several undetermined isolates were found in some population samples. In contrast to the previously reported population samples from Nepal, especially from dried soil samples, some of these populations appeared to be rather heavily loaded with mutations that are deleterious to the sexual cycle (i.e. sexual compatibility, zygospore formation and germination). By genetic analysis, a zygote maturation-defective mutation (zym) was detected. One reason for such a heavy genetic load was suggested to be that most population samples had been maintained exclusively by asexual reproduction for a long period in large lakes and nearby ponds, or left-over vegetative populations in paddy fields after other members entered into dormancy through sexual reproduction. The significance of studying such mutations at sexual gene loci is discussed in the light of speciation problems in microalgae.     

多疣狭口蛙昆明种群雌雄配对行为及形态适应选择

测量和比较采自昆明东北郊的24对抱对多疣狭口蛙(Kaloulaverrucosa)标本,同时野外观察其繁殖行为。配对雌雄蛙的体长、体重和头宽3个形态特征的Pearson相关系数大于0·5;择体长和体重作回归分析,结果表明,雌雄蛙体长选择和雌雄蛙体重配对都呈良好的线性关系。雌蛙选择雄蛙体长为自身体长的(81·8±5·7)%、体重为自身体重的(53·1±10·7)%的个体为最适配偶。雌蛙选择体长较长而体重较轻的雄蛙作为配偶,这样有利于形成较好的空间(雄性胸腹部皮肤腺长度,雌雄抱对时泄殖孔位置等)配对关系。多疣狭口蛙属于雌性选择模式,即雄蛙鸣叫,雌蛙受吸引,主动接近雄蛙,如果雄蛙条件适合则形成配对。     

浮床生态场空间分布特征

在原位试验的基础上,建立浮床生态场模型.模拟结果表明,浮床生态势值在"场源"处最大,从浮床"场源"向周围水平方向以及向水下垂直方向,随着距离的增加生态势逐渐下降.同时,浮床植物的生物量对生态场效应有显著影响,浮床植物生长越旺盛、生物量越大,生态场效应越强.1 m×1m的浮床,当植物生物量为39.40 kg·m-2时,浮床系统可最远影响到距离浮床1.86 m范围内的水体,并对浮床周围10.90 m2范围内的水体产生作用.该模型可以模拟浮床生态场的空间分布特征及其随植物生长发育的动态变化过程,并可通过模型定量化反映浮床系统在其作用范围内对任意一点的作用强度及其综合影响,该模型的构建将为生态浮床的野外应用提供理论依据.     

Morphological and cytogenetic characteristics of intergroup hybrids between closely related mating groups of the Closterium ehrenbergii species complex (Desmidiales, Chlorophyta)

Five F₁ hybrid strains were established from rare survivors in intergroup crosses between three closely related mating groups (A, B and H) of the Closterium ehrenbergii Meneghini ex Ralfs species complex. Cell sizes of these five strains studied under our standard culture conditions were compared to those of their parental stains and also to the total range of cell-size variation in each mating group. All five F₁ strains were larger in mean cell width than their parental strains. In cell length, three of them were larger than, one was the same as, and the other was intermediate between their parental strains. Their cell sizes were always larger than the range of their respective smaller parental mating group and three of them were larger than the range of their respective larger parental mating group.     

Sexual Selection and Breeding Patterns: Insights from Salmonids (Salmonidae)

Although "intrasexual selection" has been accepted as the mechanism by which males evolve elaborate secondary sexual traits which are used in aggressive contests, the importance of "intersexual selection" as a mechanism by which males have acquired exaggerated traits to display to females during courtship was less readily accepted. In spite of this scepticism, several genetic models have supported the latter idea, and many empirical studies showed that females were generally more discriminating in mate choice than males, because of differences in relative investment between sexes. Nowadays, this idea is reinforced by various concepts (parental investment, potential reproductive rate, environmental potential for polygamy...) which stress that the strength of sexual selection is related to many interdependent factors, such as mating systems, resource distribution (food, habitat, mate), life history and other ecological characteristics. The case of Salmonids is presented here to show how novel information on sexual selection has contributed to the understanding of the plasticity of breeding patterns in the context of evolutionary biology.