Multilocus half-tetrad analysis and centromere mapping in citrus: evidence of SDR mechanism for 2n megagametophyte production and partial chiasma interference in mandarin cv 'Fortune'

The genetic structure of 2n gametes and, particularly, the parental heterozygosity restitution at each locus depends on the meiotic process by which they originated, with first-division restitution and second-division restitution (SDR) being the two major mechanisms. The origin of 2n gametes in citrus is still controversial, although sexual polyploidisation is widely used for triploid seedless cultivar development. In this study, we report the analysis of 2n gametes of mandarin cv 'Fortune' by genotyping 171 triploid hybrids with 35 simple sequence repeat markers. The microsatellite DNA allele counting-peak ratios method for allele-dosage evaluation proved highly efficient in segregating triploid progenies and allowed half-tetrad analysis (HTA) by inferring the 2n gamete allelic configuration. All 2n gametes arose from the female genitor. The observed maternal heterozygosity restitution varied between 10 and 82%, depending on the locus, thus SDR appears to be the mechanism underlying 2n gamete production in mandarin cv 'Fortune'. A new method to locate the centromere, based on the best fit between observed heterozygosity restitution within a linkage group and theoretical functions under either partial or no chiasmata interference hypotheses was successfully applied to linkage group II. The maximum value of heterozygosity restitution and the pattern of restitution along this linkage group would suggest there is partial chiasma interference. The implications of such a restitution mechanism for citrus breeding are discussed.     

Genetic structure of an introduced paper wasp,Polistes chinensis antennalis (Hymenoptera,Vespidae) in New Zealand

Several eusocial wasps are prominent invaders to remote islands. The paper wasp Polistes chinensis antennalis is native to East Asia, was introduced to New Zealand in 1979 and has expanded its distribution there. This provides an excellent opportunity to examine the impacts of an initial bottleneck and subsequent expansion on genetic structure. We analysed and compared the genetic population structures of the native (Japan and South Korea) and invasive New Zealand populations. Although 94% of individuals had shared haplotypes detected across both populations, the remaining 6% had private haplotypes identified in only one of the three countries. The genetic variation at microsatellite loci was lower in New Zealand than in native countries, and the genetic structure in New Zealand was clearly distinct from that in its native range. Higher frequencies of diploid‐male‐ and triploid‐female‐producing colonies were detected in New Zealand than in the native countries, showing the reduction in genetic variation via a genetic bottleneck. At least two independent introductions were suggested, and the putative source regions for New Zealand were assigned as Kanto (central island) and Kyushu (south island) in Japan. Serial founder events following the initial introduction were also indicated. The estimated dispersal distance between mother and daughter in New Zealand was twice that in Japan. Thus, the introduction history of P. chinensis antennalis in New Zealand is probably the result of at least two independent introductions, passing through a bottleneck during introduction, followed by population expansion from the point of introduction.     

A somatic mosaic of the gynogenetic Amazon molly

Breeding experiments with the gynogenetic Amazon molly, Poecilia formosa , proved that mosaicism, detected by flow cytometry with the fluorescent dye DAPI was restricted to the soma, and that gynogenetic reproduction was maintained by the offspring. Potential pathways for how mosaicism might have arisen in P. formosa are discussed.     

A rare case of a natural contact zone in Morocco between an autopolyploid and an allopolyploid of Centaurea aspera with sterile tetraploid hybrids

A new contact zone between Centaurea aspera and Centaurea seridis was found in Morocco. Chromosome counts and flow cytometry showed that both taxa were tetraploid (4x = 44). A literature review and morphometric analysis established that C. aspera corresponds to the autopolyploid C. aspera subsp. gentilii and C. seridis corresponds to the allopolyploid C. seridis var. auriculata. This contact area was compared with the homologous contact zones in Spain formed by the diploid C. aspera subsp. stenophylla and the tetraploid C. seridis subsp. maritima. Natural hybrids between parental species were frequent in both areas. In Spain, hybrids were triploid (from reduced gametes A and gamete AB), highly sterile and exerted a ‘triploid block’. In Morocco, cytometry showed that hybrids were tetraploid and, therefore, probably fertile, but all the capitula lacked achenes. It is likely that the resulting genome of the new tetraploid hybrid (AAAB), through the fusion of reduced gametes AA (from subsp. gentilii) and AB (from var. auriculata), could explain irregularities in meiosis through formation of aneuploid gametes and, therefore, infertility of the hybrid. Moroccan sterile tetraploid hybrids develop, but have the identical irregularities to Spanish triploids, probably due to the odd number of homologous chromosomes. The new hybrid is first described as C. x subdecurrens nothosubsp. paucispinus. In addition, distribution and ecological traits are analysed.     

禾本科三倍体: 形成、鉴定与利用

禾本科三倍体的形成途径包括2n配子融合、倍性间杂交、多精受精和胚乳培养。其中, 2n配子融合和倍性间杂交分别为自然界和人工合成三倍体的主要途径。该文介绍了形态学观测、染色体分析、流式细胞术和分子标记等倍性鉴定方法在禾本科三倍体中的应用及其优缺点。目前, 三倍体在禾谷类作物中无直接应用价值, 但可作为通往多倍体、非整倍体和转移异源基因的遗传桥梁。多年生禾本科三倍体(特别是异源三倍体)在饲草或能源作物中已得到广泛应用, 在该类型禾本科作物中均可直接尝试三倍体育种。多倍体的三倍体育种和无融合生殖三倍体育种可作为未来禾本科三倍体的研究方向。三倍性胚乳培养可以一步合成三倍体, 多精受精可以实现遗传上3个不同基因组的一步融合, 在三倍体研究中应予以重视。鉴于2n配子融合、多精受精的稀有特性和倍性间杂交、胚乳培养频繁的染色体变异, 高通量三倍体鉴定技术的发展将是三倍体研究实现突破的关键。     

六倍体多星韭的发现及多星韭种内倍性组成及演化的分析

采用染色体常规制片技术,对云南西北部和中部地区11个多星韭居群进行了细胞学分析。结果显示:(1)云南西北部多星韭二倍体、四倍体混合分布的太子阁和华首门四倍体居群中发现了三倍体(2n=3x=21),核型公式分别为:2n=3x=21=[6m(2sat)+8sm]+(3m+4sm)和2n=3x=21=[4m(2sat)+10sm]+(2m+4sm+lst)。三倍体含2条随体染色体,它的21条染色体含2个同源染色体组和一个与它们同源性稍差的染色体组。(2)在云南中部的多星韭四倍体居群中首次发现了六倍体,核型公式为2n=6x=42=15m(3sat)+27sm,核型类型2A。(3)结合核型特征及地理分布对多星韭三倍体和六倍体的成因进行分析,提示多星韭三倍体来源于二倍体与四倍体的杂交,六倍体是通过四倍体产生的未减数配子(n=4x=28)与正常减数配子(n=2x=14)的结合形成的。本研究结果支持多星韭的分化中心是云南西北部高海拔地区的观点,并结合前人的研究和本工作的结果,对多星韭种内的倍性组成和演化进行了分析和讨论。     

造纸废水灌溉对毛白杨苗木生长及养分状况的影响

为了探讨工业造纸废水用于杨树人工林灌溉的可行性,以三倍体毛白杨(triploid Populus tomentosa)1年生盆栽苗木为对象,研究了不同浓度造纸废水(分别稀释到12.5％ (IF7Q)、16.7％ (1F5Q)、25％(1F3Q)、33.3％ (1F2Q)、50％ (1F1Q))灌溉对苗木生长及养分状况、土壤化学性质的影响.结果表明:造纸废水灌溉对土壤pH值、速效P含量无显著影响(P＞0.05),但能显著提高土壤有机质、全N及碱解N的含量(P＜0.05).适当稀释的废水灌溉能促进三倍体毛白杨的苗木生长,提高土壤和植株养分水平:灌溉后1F5Q地径、苗高生长量分别为10.5 mm和97.3 cm,较CK分别显著增加102％和47％ (P＜0.05);1F5Q和1F3Q处理苗木总生物量为247 g和230 g,分别较CK显著提高19％和11％(P＜0.05);废水灌溉可显著提高植株根、叶N含量和茎P含量(P＜0.05),但对植株叶、根P含量和茎N含量影响不大(P＞0.05).造纸废水通过一定处理后,可应用于苗木灌溉并促进其生长,提高地力.对于三倍体毛白杨,将废水稀释到16％-25％能起到较好的灌溉效果.     

Microsatellite and flow cytometry analysis to help understand the origin of Dioscorea alata polyploids

Background and Aims Dioscorea alata

is a polyploid species with a ploidy level ranging from diploid (2n = 2x = 40) to tetraploid (2n = 4x = 80). Ploidy increase is correlated with better agronomic performance. The lack of knowledge about the origin of D. alata spontaneous polyploids (triploids and tetraploids) limits the efficiency of polyploid breeding. The objective of the present study was to use flow cytometry and microsatellite markers to understand the origin of D. alata polyploids.

Methods

Different progeny generated by intracytotype crosses (2x × 2x) and intercytotype crosses (2x × 4x and 3x × 2x) were analysed in order to understand endosperm incompatibility phenomena and gamete origins via the heterozygosity rate transmitted to progeny.

Results

This work shows that in a 2x × 2x cross, triploids with viable seeds are obtained only via a phenomenon of diploid female non-gametic reduction. The study of the transmission of heterozygosity made it possible to exclude polyspermy and polyembryony as the mechanisms at the origin of triploids. The fact that no seedlings were obtained by a 3x × 2x cross made it possible to confirm the sterility of triploid females. Flow cytometry analyses carried out on the endosperm of seeds resulting from 2x × 4x crosses revealed endosperm incompatibility phenomena.

Conclusions

The major conclusion is that the polyploids of D. alata would have appeared through the formation of unreduced gametes. The triploid pool would have been built and diversified through the formation of 2n gametes in diploid females as the result of the non-viability of seeds resulting from the formation of 2n sperm and of the non-viability of intercytotype crosses. The tetraploids would have appeared through bilateral sexual polyploidization via the union of two unreduced gametes due to the sterility of triploids.     

Introgression and phylogeography of Betula nana (diploid), B. pubescens (tetraploid) and their triploid hybrids in Iceland inferred from cpDNA haplotype variation

Aim The objective was to find direct genetic evidence supporting introgressive hybridization between tetraploid tree birch (Betula pubescens) and diploid dwarf birch (B. nana), via triploid hybrids, and to investigate an association between the introgression and phylogeographical distribution of Icelandic birch. Location Samples were collected from 463 trees in 12 woodlands in Iceland and eight locations in Norway, Sweden, Scotland and Greenland. Methods Ploidy status of individual trees was determined by chromosome counting. Variation in the chloroplast genome was assessed using polymerase chain reaction‐restriction fragment length polymorphism. The geographical distribution of the haplotypes was mapped. The haplotype variation and introgression ratios (IG) were analysed statistically. Results Thirteen haplotypes were identified among Icelandic samples. The most common haplotype (T, 49% occurrence) was present in all ploidy groups and in all woodlands. All common haplotypes were shared between the triploid group and the parental species, indicating introgressive hybridization. This was supported by the statistical analysis of IG indices and the variation components. Considerable differences existed among samples, shaped by isolation by distance and local introgression. An east–west phylogeographical distribution in Iceland was observed. Main conclusions Despite extensive introgression across species and ploidy levels, a biogeographical pattern has been observed, and this may indicate different population histories or multiple origins of Icelandic birch. The chloroplast haplotype diversity found in Iceland resembles that found in birch populations from northern Scandinavia.