Population genetic structure and differentiation of Anthoxanthum alpinum in the subalpine-alpine ecocline of Swiss Alps

Allozyme variation in 6 enzyme systems coding 10 loci was estimated for 18 subpopulations of Anthoxanthum alpinum from three altitudinal transects in two localities of the Swiss Alps. Mean proportions of polymorphic loci (95% criterion), average number of alleles per locus, and mean expected heterozygosity were 64.9%, 2.37 and 0. 252, respectively. Mean genetic distance among populations was 0.011, and 79% of the genetic variation resided within populations. Based on allozyme analysis, marginal subpopulations appeared to have similar level of genetic variability to central subpopulations. Relatively high genetic differentiation, low gene flow values and small neighbourhood sizes suggested that inbreeding followed by genetic drift was possible causes of lowgenetic variability in Arpette A. alpinum populations.     

阿尔卑斯山黄花茅(Anthoxanthum alpinum)沿海拔梯度的基因频率分布

在瑞士阿尔卑斯山的2个定点样地,黄花茅从森林线（海拔1700 m）到山顶（海拔2830 m）呈连续分布。在海拔梯度样带的最高和最低定样场所间的垂直距离差不多为1000 m,但2个定样场所间相距仅仅1.4 km。在所研究的3个海拔梯度样带中,3个同工酶位点（Px_1, Got_2和 Mdh_1）被观察到有统计意义的倾群变异。研究结果显示：沿海拔梯度样带的亚居群间的基因流可能太弱不足以克服自然选择的影响,后者促使对局部环境的适应。在这种情况下,温度因子至少可能作为一种主要的自然选择力起作用。     

Subpopulation differentiation of Anthoxanthum alpinum (Poaceae) along an altitudinal gradient detected by random amplified polymorphic DNA

Random amplified polymorphic DNA (RAPD) phenotypes generated by 15 primers were scored in 5 subpopulatious of Anthoxanthum alpinum along an altitudinal gradient. Although few subpopulation-specific markers were found, a significant population differentiation was revealed by the use of principal factor analysis and Ward' s cluster analysis. The result was also confirmed by the significant positive correlation when comparing beth the matrix of similarity between the individuals within subpopulations and the matrix among subpopulations, as well as the matrix of distances and the differences of altitude between the subpopulations (Mantel test). This study suggests that the differences of altitude between subpopulation sites may result in the differences of phenologicalperiod for flowering and growth which restrict gene flow. Decontaminated thianthrene disproportion. Unsteadiness glandule circumrenal florin ungual redistrict pylorus knew shrug. 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Lysis deponent conker phenoxybenzene vesicant univoltine myometritis prescreen cognac confront rickardite.      

阿尔卑斯山高山-亚高山过渡区高山黄花茅的群体遗传结构和分化研究

应用等位酶分析,在瑞士阿尔卑斯山的阿尔拜特(Arpette)和拜阿尔普(Belalp),沿三个不同的海拔梯度,研究了高山黄花茅3个自然居群的遗传变异和分化。研究结果表明,平均的多态性位点比例为64．9％,每个位点平均等位基因数为2．37,平均期望杂合性为0．252。亚居群间平均的遗传距离为0．011,发现79％的遗传变异存在于居群内。基于等位酶分析,边缘亚居群与中心亚居群似乎有类似的遗传变异性。相对比较高的遗传分化、低的亚居群间基因流和小的邻居大小值暗示,近交和随后的遗传漂变可能是导致阿尔拜特黄花茅居群遗传变异性较低的主要原因。     

Pollen-wall formation in Arum alpinum

BACKGROUND AND AIMS: Arum alpinum has a quite uncommon pollen wall. A sporopolleninous ektexine is missing. The outermost pollen wall layer is formed by the endexine which is covered by polysaccharidic ornamentation elements. An ontogenetical investigation was accomplished to clarify pollen-wall development, with special reference to callose and pollen-wall development. METHODS: Plants of Arum alpinum grown in their natural habitat were collected once a week within the vegetative period and processed for semi- and ultra-thin sectioning. KEY RESULTS: At any stage of pollen-wall formation callose is missing. Microspores are released from the tetrad by invagination of the amoeboid tapetum. The polysaccharidic wall ornamentations are formed by the tapetum. CONCLUSIONS: There appears to be no truth in the dogma that callose is essential for microspore separation and release from the tetrad. The lack of callose does not influence fertility but could be the reason for the uncommon pollen wall, where a sporopolleninous ektexine is missing.     

Differentiating diploid and triploid individuals using single nucleotide polymorphisms genotyped by amplicon sequencing

Triploidy can occur naturally or be induced in fish and shellfish during artificial propagation in order to produce sterile individuals. Fisheries managers often stock these sterile triploids as a means of improving angling opportunities without risking unwanted reproduction of the stocked fish. Additionally, the rearing of all‐triploid individuals has been suggested as a means to reduce the possibility of escaped aquaculture fish interbreeding with wild populations. Efficient means of determining if an individual is triploid or diploid are therefore needed both to monitor the efficacy of triploidy‐inducing treatments and, when sampling fish from a body of water that has a mixture of diploids and triploids, to determine the ploidy of a fish prior to further analyses. Currently, ploidy is regularly measured through flow cytometry, but this technique typically utilizes a fresh blood sample. This study presents an alternative, cost‐effective method of determining ploidy by analysing amplicon‐sequencing data for biallelic single‐nucleotide polymorphisms (SNPs). For each sample, heterozygous genotypes are identified and the likelihoods of diploidy and triploidy are calculated based on the read counts for each allele. The accuracy of this method is demonstrated using triploid and diploid brook trout (Salvelinus fontinalis) genotyped with a panel of 234 SNPs and Chinook salmon (Oncorhynchus tshawytscha) genotyped with a panel of 298 SNPs following the GT‐seq methodology of amplicon sequencing.     

白杨不同倍性花粉的辐射敏感性及其应用

通过对白杨二倍性未减数花粉和单倍性花粉萌发率和萌发进程的观察统计,证明不同 倍性花粉对（６０）ＣＯγ射线辐射的敏感性不同,其中单倍性花粉的敏感性约为二倍性花粉的 ２倍。 为克服二倍性花粉在参与受精过程中萌发迟缓、竞争能力较差的问题,可利用这种花粉倍性 对辐射的敏感性差异,施加一定剂量的辐射以刺激二倍性花粉萌发,同时抑制或杀死部分单 倍性花粉,从而在一定程度上相对提高柱头上萌发和萌发较快的二倍性花粉比率。三倍体育 种结果亦证明了花粉辐射在提高三倍体得率方面应用的有效性,其中在银腺杨×毛新杨组 合中,以 １４７０ｒａｄ处理最佳,三倍体得率达３．８％,对照未检出三倍体;而在施加 １６８０ｒａｄ的银 腺杨×毛新杨杂交组合中,则获得了高达 １２．９％的三倍体。     

QTL mapping in outbred tetraploid (and diploid) diallel populations

Over the last decade, multiparental populations have become a mainstay of genetics research in diploid species. Our goal was to extend this paradigm to autotetraploids by developing software for quantitative trait locus (QTL) mapping in connected F1 populations derived from a set of shared parents. For QTL discovery, phenotypes are regressed on the dosage of parental haplotypes to estimate additive effects. Statistical properties of the model were explored by simulating half-diallel diploid and tetraploid populations with different population sizes and numbers of parents. Across scenarios, the number of progeny per parental haplotype (pph) largely determined the statistical power for QTL detection and accuracy of the estimated haplotype effects. Multiallelic QTL with heritability 0.2 were detected with 90% probability at 25 pph and genome-wide significance level 0.05, and the additive haplotype effects were estimated with over 90% accuracy. Following QTL discovery, the software enables a comparison of models with multiple QTL and nonadditive effects. To illustrate, we analyzed potato tuber shape in a half-diallel population with three tetraploid parents. A well-known QTL on chromosome 10 was detected, for which the inclusion of digenic dominance lowered the Deviance Information Criterion (DIC) by 17 points compared to the additive model. The final model also contained a minor QTL on chromosome 1, but higher-order dominance and epistatic effects were excluded based on the DIC. In terms of practical impacts, the software is already being used to select offspring based on the effect and dosage of particular haplotypes in breeding programs.     

二倍体和四倍体栽培苦荞的细胞学比较研究

观察和统计了二倍体和四倍体苦荞 (Fagopyrum tataricum)花粉母细胞减数分裂过程 TI、T 时期的异常程度及花粉粒败育性等多个指标 ,并比较分析了二倍体和四倍体苦荞在这些指标上的差异显著性。结果显示 :四倍体苦荞的异常 TI细胞频率、异常 T 细胞频率、TI细胞平均微核数、T 细胞平均微核数及花粉败育率均显著高于二倍体苦荞 ,这表明四倍体苦荞的遗传稳定性和花粉可育性都比二倍体苦荞差。     

Ecological and evolutionary importance of neighbors in the grass Anthoxanthum odoratum

Summary A field experiment was performed to estimate the relative importance of neighbors and the rest of the environment for the growth, mortality and reproductive output of cloned individuals of the perennial bunchgrass Anthoxanthum odoratum. Single cloned Anthoxanthum tillers (targets) were reciprocally transplanted between a xeric and a mesic grassland site with one of four neighbor treatments: (1) no neighbors, (2) Anthoxanthum neighbors transplanted from the xeric site, (3) Anthoxanthum neighbors from the mesic site, and (4) Holcus lanatus neighbors. Targets without neighbors had a twofold higher two year reproductive output (RO) than those with neighbors, but there was no difference among neighbor treatments. No overall site effect on two year RO was found, because the site with the highest mortality among targets produced larger plants, with more inflorescences. Neighborhood competition was more intense at the xeric site than at the mesic site. The effects of environmental and neighborhood variation on Anthoxanthum were additive, rather than interactive. Population origin did not affect target performance significantly. Anthoxanthum neighbors of different origin did respond differentially to transplant site. There was a strong target genotype x site interaction, but no genotype x neighborhood interaction.     

Pollen and stomata morphometrics and polyploidy in Eriotheca (Malvaceae‐Bombacoideae)

Approximately 70% of the angiosperm species are polyploid, an important phenomenon in the evolution of those plants. But ploidy estimates have often been hindered because of the small size and large number of chromosomes in many tropical groups. Since polyploidy affects cell size, morphometric analyses of pollen grains and stomata have been used to infer ploidy level. Polyploidy is present in many species of the Cerrado, the Neotropical savanna region in Central Brazil, and has been linked to apomixis in some taxa. Eriotheca gracilipes and Eriotheca pubescens are common tree species in this region, and present cytotypes that form reproductive mosaics. Hexaploid individuals (2n = 6x = 276) are polyembryonic and apomictic, while tetraploid and diploid individuals (2n = 2x = 92, 2n = 4x = 184) are sexual and monoembryonic. We tested whether morphometric analysis can be used to estimate ploidy levels in E. gracilipes and E. pubescens individuals. Pollen material from diploid and hexaploid individuals of E. gracilipes, and tetraploid and hexaploid individuals of E. pubescens, were fixed in 50% FAA, and expanded leaves were dried in silica gel. Pollen grains and stomata of at least five individuals from each population were measured. The results demonstrate that all measures were significantly different among cytotypes. Individuals with higher levels of ploidy (hexaploid) all presented measurements that were higher than those with lower levels (diploid and tetraploid). There was no overlap between ploidy levels in each species at 95% confidence interval. Thus, the size of the pollen grains and stomata are effective parameters for analysis of ploidy levels in E. gracilipes and E. pubescens.     

卷丹小孢子母细胞减数分裂染色体行为的研究

对引种于陕西汉中地区的三倍体卷丹的小孢子母细胞减数分裂进程、花粉生活力和可育性进行了分析。结果显示:减数分裂时期与花蕾、花药长度及花药壁颜色呈现出一定的相关性,处于减数分裂时期的卷丹花蕾长度在2.5～3.2 cm之间,花药长度在1.4～1.8 cm之间;从细线期到成熟花粉粒,花药壁的颜色呈现出淡绿色-淡黄绿色-黄绿色-橘黄色的变化趋势,花药内容物颜色也从透明逐渐变为粘稠的橘红色。减数分裂终变期染色体构型以三价体为主,平均构型为:11.6Ⅲ+2Ⅱ+1.1Ⅰ,可判断该种源地卷丹为同源三倍体。减数分裂中出现染色体不均等分裂、染色体落后、染色体桥及微核现象,同时观察到成熟花粉粒大小差异较大,极轴与赤道轴乘积在839.465～3327.907μm2范围内,趋于正态分布,为三倍体卷丹花粉的高度不育提供了细胞学证据。以卷丹为母本与大花卷丹进行杂交,得到一定数量的种子,其中有胚率达到了43.0%,成苗率为67.4%,说明三倍体卷丹在百合育种中具有一定的价值。     

Reproductive ecology of the endangered Alpine species Eryngium alpinum L. (Apiaceae): phenology, gene dispersal and reproductive success

BACKGROUND AND AIMS: Eryngium alpinum (Apiaceae) is an endangered perennial, characteristic of the Alpine flora. Because the breeding system influences both demographic (reproductive success) and genetic (inbreeding depression, evolutionary potential) parameters that are crucial for population maintenance, the reproductive ecology of E. alpinum was investigated. Specifically, the aims of the study were (1) to determine the factors (resources and/or pollen) limiting plant fitness; and (2) to assess the potential for gene flow within a plant, within a patch of plants, and across a whole valley where the species is abundant. METHODS: Field experiments were performed at two sites in the Fournel valley, France, over three consecutive years. Studies included a phenological survey, observations of pollinators (visitation rates and flight distances), dispersal of a fluorescent powder used as a pollen analogue, the use of seed traps, determination of the pollen/ovule ratio, and an experiment to test whether seed production is limited by pollen and/or by resources. KEY RESULTS: E. alpinum is pollinated by generalist pollinators, visitation rates are very high and seed set is resource- rather than pollen-limited. The short flights of honeybees indicate a high potential for geitonogamy, and low pollen and seed dispersals suggest strong genetic structure over short distances. These results are interpreted in the light of previous molecular markers studies, which, in contrast, showed complete outcrossing and high genetic homogeneity. CONCLUSION:S. The study highlights the usefulness of adopting several complementary approaches to understanding the dynamic processes at work in natural populations, and the conservation implications for E. alpinum are emphasized. Although the studied populations do not seem threatened in the near future, long-term monitoring appears necessary to assess the impact of habitat fragmentation. Moreover, this study provides useful baseline data for future investigations in smaller and more isolated populations.     

Phylogeography of the endangered Eryngium alpinum L. (Apiaceae) in the European Alps

We studied the phylogeography of Eryngium alpinum by sequencing two intergenic chloroplast spacers, trnH-psbA and trnS-trnG (1322 bp). The sampling design included 36 populations and 397 individuals spanning the entire distribution range of the species, from France to Bosnia. Twenty-one haplotypes were characterized and polymorphism was observed both within and among populations. Population differentiation was strong (F(ST) = 0.92) and largely explained by the distinction of five geographic regions: Southwestern, Western, Middle, Eastern Alps and Balkans (F(CT) = 0.62). Moreover, N(ST) was significantly higher than G(ST) (P < 0.05), showing the existence of a phylogeographic pattern. Six major lineages were recognized using samova and median-joining networks. One lineage, highly divergent from the other ones, was only found in the Balkans and probably persisted in situ during last glaciations. All other lineages might have survived in a Southwestern refugium (Mercantour) and colonized the entire Alpine arc (Southwestern, Western, Middle and Eastern Alps) through repeated colonization events at different time periods. This is the first empirical study suggesting Southern refugia for calcareous Alpine plants, although the existence of a secondary refugium in northern Italy/Austria is also suspected. We also observed recent haplotype diversification, especially in the Southwestern Alps.     

桔梗花粉萌发与花粉管生长研究

以2年生桔梗植株为材料,采用液体培养法研究了培养基种类、PEG、蔗糖、pH以及培养温度、培养时间对桔梗花粉离体萌发生长的影响,结果表明:(1)浓度为100~150 g.L-1的PEG可显著促进桔梗花粉萌发和花粉管的生长;200~250 g.L-1PEG显著促进花粉萌发,但对花粉管生长的作用不显著。(2)100 g.L-1的蔗糖有利于花粉萌发和花粉管生长,高浓度蔗糖(200 g.L-1)有明显抑制作用;(3)桔梗花粉离体萌发和花粉管生长的适宜培养基为ME3+BK+10%蔗糖+150 g.L-1PEG(pH5.8);(4)25~40℃条件下桔梗花粉均可较好萌发,以30℃培养1.5 h为最佳培养条件。     

Artificial Germination of Gossypium Pollen and Its utilization in Interspecific Fertilization In Vitro

The effects of different media and cultural methods on cotton pollen germination were compared. Pollen germination was successful on the improved Taylor solid medium in which the extracts of the stigmata and petals of the original species, and the comprehensive amino acids were added, respectively. The germinating pollens were viable, and active protoplasmic streaming was observed within the pollen tube. It was found that low temperature pretreatment could increase the pollen germination rate. The reasons that low temperature pretreatment stimulated pollen germination and castor oil caused abortive germination of cotton pollens were discussed. In vitro fertilization between two species of Gossypium can be promoted by smearing the stigmata with the improved Taylor medium.     

Phenotypic plasticity and population differentiation in seeds and seedlings of the grass Anthoxanthum odoratum

Summary Seeds of Anthoxanthum odoratum were transplanted reciprocally between a xeric and a mesic field population that were genetically differentiated in adult traits. In one experiment seeds were reciprocally buried in bags in the soil, in a second experiment seeds were reciprocally sown in small plots. For most traits, site effects were much larger than seed-source effects. Germination, emergence, mortality of buried seed and recruitment were significantly higher at the xeric site than at the mesic site, irrespective of population of origin. Seed dormancy, was significantly higher for seed originating from the mesic than from the xeric population. Seedling recruits originating from the xeric population tended to be larger at both sites. Fecundity of seedling recruits depended on the environment; fecundities of plants growing in the xeric site had more than double the fecundity of plants growing in the mesic site. Phenotypic plasticity rather than population differences determined variation in performance in the seed and seedling stages.     

Spermatozoa production by triploid males in the New Zealand freshwater snail Potamopyrgus antipodarum

Asexual lineages derived from dioecious taxa are typically assumed to be all female. Even so, asexual females from a variety of animal taxa occasionally produce males. The existence of these males sets the stage for potential gene flow across asexual lineages as well as between sexual and asexual lineages. A recent study showed that asexual triploid female Potamopyrgus antipodarum, a New Zealand freshwater snail often used as a model to study sexual reproduction, occasionally produce triploid male offspring. Here, we show that these triploid male P. antipodarum (1) have testes that produce morphologically normal sperm, (2) make larger sperm cells that contain more nuclear DNA than the sperm produced by diploid sexual males, and (3) produce sperm that range in DNA content from haploid to diploid, and are often aneuploid. Analysis of meiotic chromosomes of triploid males showed that aberrant pairing during prophase I probably accounts for the high variation in DNA content among sperm. These results indicate that triploid male P. antipodarum produce sperm, but the extent to which these sperm are able to fertilize female ova remains unclear. Our results also suggest that the general assumption of sterility in triploid males should be more closely examined in other species in which such males are occasionally produced. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 110 , 227–234.