Habitat preference and flowering‐time variation contribute to reproductive isolation between diploid and autotetraploid Anacamptis pyramidalis

Tetraploid lineages are typically reproductively isolated from their diploid ancestors by post‐zygotic isolation via triploid sterility. Nevertheless, polyploids often also exhibit ecological divergence that could contribute to reproductive isolation from diploid ancestors. In this study, we disentangled the contribution of different forms of reproductive isolation between sympatric diploid and autotetraploid individuals of the food‐deceptive orchid Anacamptis pyramidalis by quantifying the strength of seven reproductive barriers: three prepollination, one post‐pollination prezygotic and three post‐zygotic. The overall reproductive isolation between the two cytotypes was found very high, with a preponderant contribution of two prepollination barriers, that is phenological and microhabitat differences. Although the contribution of post‐zygotic isolation (triploid sterility) is confirmed in our study, these results highlight that prepollination isolation, not necessarily involving pollinator preference, can represent a strong component of reproductive isolation between different cytotypes. Thus, in the context of polyploidy as quantum speciation, that generates reproductive isolation via triploid sterility, ecological divergence can strengthen the reproductive isolation between cytotypes, reducing the waste of gametes in low fitness interploidy crosses and thus favouring the initial establishment of the polyploid lineage. Under this light, speciation by polyploidy involves ecological processes and should not be strictly considered as a nonecological form of speciation.     

Polymorphic microsatellite markers in polyploid Lepidium draba L. ssp. draba (Brassicaceae) and cross‐species amplification in closely related taxa

Heart‐podded hoary cress, Lepidium draba L. ssp. draba (Brassicaceae) is a noxious invasive weed in the USA. At present, efficient biological control of this Eurasian native weed in the USA is hampered by lack of knowledge of its population genetic structure and colonization process. Here, we describe the development of 11 polymorphic microsatellite markers that also reveal the polyploidy of this weed. Successful cross‐species amplification highlights the possibility of using these markers for genetic studies in other closely related species.     

秋水仙素结合组织培养技术诱导大葱多倍体的研究

该文研究了秋水仙素不同浓度不同时间处理大葱愈伤组织的诱导效果,发现随着处理时间和浓度的增加,愈伤组织死亡率增加,多倍体细胞诱导率在一定范围内随处理时间和浓度增加升高,但浓度超过一定量诱导率则相反,以0.06%的秋水仙素处理72h多倍体细胞诱导率最佳,且不对愈伤组织产生严重伤害,获得的变异材料与正常二倍体相比,其叶片变粗,生长迟缓,气孔器变大,经鉴定为多倍体。     

Identification of SNP markers for inferring phylogeny in temperate bamboos (Poaceae: Bambusoideae) using RAD sequencing

Phylogenetic relationships among temperate species of bamboo are difficult to resolve, owing to both the challenge of detecting sufficiently variable markers and their polyploid history. Here, we use restriction site–associated DNA sequencing to identify candidate loci with fixed allelic differences segregating between and within two temperate species of bamboos: Arundinaria faberi and Yushania brevipaniculata. Approximately 27 million paired‐end sequencing reads were generated across four samples. From pooled data, we assembled 67 685 and 70 668 de novo contigs from partial overlap among paired‐end reads, with an average length of 240 and 241 bp for the two species, respectively, which were used to investigate functional classification of RAD tags in a blastx search. Analysed separately by population, we recovered 29 443 putatively orthologous RAD tags shared across the four sampled populations, containing 28 023 sequence variants, of which c. 13 000 are segregating between species, and c. 3000 segregating between populations within each species. Analyses based on these RAD tags yielded robust phylogenetic inferences, even with data set constructed from surprisingly few loci. This study illustrates the potential for reduced‐representation genome data to resolve difficult phylogenetic relationships in temperate bamboos.     

Assignment of SNP allelic configuration in polyploids using competitive allele-specific PCR: application to citrus triploid progeny

Background

Polyploidy is a major component of eukaryote evolution. Estimation of allele copy numbers for molecular markers has long been considered a challenge for polyploid species, while this process is essential for most genetic research. With the increasing availability and whole-genome coverage of single nucleotide polymorphism (SNP) markers, it is essential to implement a versatile SNP genotyping method to assign allelic configuration efficiently in polyploids.

Scope

This work evaluates the usefulness of the KASPar method, based on competitive allele-specific PCR, for the assignment of SNP allelic configuration. Citrus was chosen as a model because of its economic importance, the ongoing worldwide polyploidy manipulation projects for cultivar and rootstock breeding, and the increasing availability of SNP markers.

Conclusions

Fifteen SNP markers were successfully designed that produced clear allele signals that were in agreement with previous genotyping results at the diploid level. The analysis of DNA mixes between two haploid lines (Clementine and pummelo) at 13 different ratios revealed a very high correlation (average = 0·9796; s.d. = 0·0094) between the allele ratio and two parameters [θ angle = tan⁻¹ (y/x) and y′ = y/(x + y)] derived from the two normalized allele signals (x and y) provided by KASPar. Separated cluster analysis and analysis of variance (ANOVA) from mixed DNA simulating triploid and tetraploid hybrids provided 99·71 % correct allelic configuration. Moreover, triploid populations arising from 2n gametes and interploid crosses were easily genotyped and provided useful genetic information. This work demonstrates that the KASPar SNP genotyping technique is an efficient way to assign heterozygous allelic configurations within polyploid populations. This method is accurate, simple and cost-effective. Moreover, it may be useful for quantitative studies, such as relative allele-specific expression analysis and bulk segregant analysis.     

中国蔷薇属6个种的染色体研究

本文对原产中国的蔷薇属6种植物(Rosa spp.)进行了染色体观察,其中1种(巨花蔷薇(Rosa gigantea(Crep)Rehd.et Wils.))为国家重点保护植物,2种(疏花蔷薇(R.laxa Retz.)、宽刺蔷薇(R.platyacantha Shrenk))为国内首次报道。观察结果如下:染色体数目为2n=2x=14或2n=3x=21,均为小型染色体(2.24—2.78μm),其中大部分长度接近,染色体长度之比小于2;属对称核型。文中讨论了一些种(变种)的染色体数目及核型。