在六倍体小麦的异源多倍化早期微卫星侧翼序列迅速发生了改变

在异源多倍体形成的早期, DNA序列和基因的表达迅速发生了改变. 以异源六倍体小麦为例, 比较了四倍体小麦与节节麦合成六倍体小麦前后, 位于普通小麦D染色体组不同染色体臂上的特异性引物揭示的微卫星位点变化特点. 结果表明, 从四倍体小麦与节节麦杂交, 将A, B与D染色体组结合在一起并加倍得到AABBDD的六倍体小麦这一“剧烈事件”中: (ⅰ) 微卫星的侧翼序列发生了变化, 导致出现了供体物种没有的新带纹或供体物种的带纹消失, 其中, 供体物种的带纹消失是主要的. (ⅱ) 供体物种的带纹消失不是随机的, 而是四倍体小麦消失频率远高于节节麦的频率, 即发生在A, B染色体组的消失频率比发生在D染色体组的频率高得多. (ⅲ) 微卫星侧翼序列的变化在多倍化的早期(F₁代或S₁代)就开始发生. 由此看来, 微卫星两边的侧翼区域在多倍化过程中很活跃, 是容易发生变化的区域. 微卫星的生物学功能可能与多倍体进化过程有关, 微卫星两边的侧翼区域在多倍化过程的早期迅速发生有方向性的改变可能有利于新形成异源多倍体的迅速进化, 从而使不同染色体组在遗传上迅速达到协调.     

问：秋水仙素处理单倍体得到的一定是纯合子吗？

答：单倍体是指体细胞中含有本物种配子染色体数目的个体。那么如果此物种是二倍体如某玉米植株基因型是Aa．其单倍体植株的基因型为A、a两种,当用秋水仙素加倍以后．生成植株的基因型为AA、aa两种纯合子;但如果此物种为四倍体、六倍体、八倍体等偶数倍多倍体时,如某马铃薯植株基因型为从aa时,则形成的单倍体植株的基因型为AA、Aa、aa3种类型。     

使用染色体数目符号的建议

我们感到目前使用染色体数目的符号不够准确,常常看到一些文章、小册子是这样叙述染色体数目的。 “以”表示基本染色体组的染色体数,称为单倍体,则含有二个染色体组的细胞或个体称为二倍体,用2n表示;含有三个染色体组的细胞或个体称为三倍体,用3n表示;依此类推,还有四倍体、五倍体、六倍体……可用4n、5n、6n……表示。凡体细胞中具有二倍以上的染色体数的生物均称为多倍体,包括三倍体、四倍体、五倍体……等”。 “小麦属的多倍体系有一粒小麦,体细胞染色体数2n=14;二粒小麦,体细胞染色体数2n=28,为异源四倍体;普通小麦,体细胞染色体数2n=42,为异源六倍体”。 这种说法并不少见。开始“以n代表基本染色体     

GENOME SIZE

采用Feulgen-显微分光光度计方法,以鸡红细胞为标准DNA（3.22pg/2C）测定了长江白鲟、达氏鲟、中华鲟、史氏鲟和北美匙吻鲟的体细胞基因组大小（DNA含量）。结果表明,上述五种鲟鱼DNA含量分别为4.11、8.26、9.07、6.07和3.96pg。长江白鲟和北美匙吻鲟均属于四倍体鱼类,分布在长江水系的中华鲟和达氏鲟两种鲟属鱼类为八倍体类型。史氏鲟初步判断为八倍体,据分析可能存在四倍体的类型。根据所得到的结果并结合已发表的DNA含量和染色体资料,分析了鲟形目鱼类在细胞遗传上的进化特点。鲟形目鱼类全为多倍体起源的鱼类,在鲟形目的两科（白鲟科和鲟科）六个属中,除鲟属外,其他五个属均为四倍体,而鲟属已发现有四倍体、八倍体、十二倍体和十六倍体。说明在细胞水平的染色体加倍与鲟形目鱼类各科、属分化没有直接联系,只在鲟形目形成的早期和鲟形目形成后的鲟属种间分化中扮演着重要角色。鲟属鱼类的染色体剧烈分化可能是造成现今鲟属种类数最多（占整个鲟形目鱼类的65%以上）的重要原因。     

动物多倍体的遗传和进化

早在1925年,Muler认为动物多倍体的性不平衡,不可能使两性多倍体作为自然界的实体在动物中存在之后,一些学者也认为动物多倍体在进化中的地位并不重要,然而,近二十年来,从无脊椎动物到脊椎动物都发现了许多多倍体的物种,有孤雌生殖的三倍体、四倍体,也有两性生殖的三倍体、四倍体、六倍体和八倍体。在昆虫和鱼类中,甚至存在着多倍体的科、属。已发现的多倍体动物,在适应性等方面远胜于它们二倍体祖先,有的甚至取代和消灭了它的祖先。动物多倍体的存在受到了生物学家     

葱属根茎组8种21居群植物的核型研究

对葱属根茎组Allium sect.Rhiziridium的8种21个地方居群的核型进行研究,以期为解决该组的种间亲缘关系和物种进化机制提供依据。贺兰韭A.eduardii和阿拉善韭A．flavovirens2个种的核型以及辉韭A．strictum的六倍体核型均属首次报道。研究结果表明：贺兰韭A.eduardii、阿拉善韭A.tlavovirens、北韭A.lineare、蒙古韭A.mongolicum和滩地韭A．oreoprasum的各居群均为二倍体,核型类型为Stebbins的2A型;韭A．tuberosum和野韭A.ramosum的各个居群均为四倍体,核型类型为2A型：辉韭A．Jtrictum的4个居群均为六倍体,核型类型为2B型。通过研究可以得出如下推论：（1）该组植物中存在着大量的多倍体或多倍体系列,染色体数目变化与物种进化具有密切相关性,多倍化可能是根茎组植物核型进化的重要机制之一;（2）随体染色体多为st或t染色体,均位于短臂末端;（3）可以认为辉韭是以增加倍性来克服该物种扩大新的生存空间所带来的困难;（4）现今栽培的韭可能是由野生的二倍体韭和四倍体韭经过长期人工驯化而来的,现今栽培的三倍体韭可能是二倍体韭和四倍体韭杂交而来,并且以无性繁殖方式保存三倍体类群的存在。     

基于高通量测序的达氏鲟微卫星标记筛选

达氏鲟(Acipenser dabryanus)是我国特有的珍稀鱼类,主要分布在长江上游干支流。由于人类活动的影响,其种群数量急剧下降,被列为国家Ⅰ级重点保护动物、IUCN红色名录极危(CR)物种。本研究针对当前达氏鲟群体的濒危现状,应用微卫星标记开展达氏鲟保护遗传学研究。通过高通量测序成功筛选出25个微卫星位点,采用18尾野生个体和30尾人工繁殖个体对其多样性进行了检验,结果表明,25个位点共检测出181个等位基因,每个位点的等位基因数(A)为4~11(平均值7.2),观测杂合度(HO)为0.160~1.000(平均值0.744),期望杂合度(HE)为0.346~0.875(平均值0.727)。所有位点均不偏离"Hardy-Weinberg"平衡(P0.05),各位点间也无连锁不平衡现象。除了其中一个位点以外,所有位点的多态信息含量(PIC)均大于0.5,其多态性较好。本次筛选的微卫星位点将有助于达氏鲟资源保护和群体遗传研究。     

不同倍性柳枝稷45S rDNA的染色体定位研究

以3个四倍体和6个八倍体栽培品种的根尖为材料,利用荧光原位杂交技术,在核型分析的基础上,开展了不同倍性柳枝稷45S rDNA的染色体定位研究。研究结果表明,四倍体柳枝稷核型公式为2n=4x=36=32m(SAT)+4sm,且45S rDNA在四倍体柳枝稷染色体上分布稳定,位于3号染色体顶端。八倍体柳枝稷栽培品种以及同一栽培品种不同个体间45S rDNA信号分布位置和数目差异较大,可大致分为四类:第Ⅰ类为较强的45S rDNA信号分别分布于染色体两臂的顶端;第Ⅱ类为较强的45S rDNA信号位于染色体一臂内部,第Ⅲ类为较强的45S rDNA信号位于染色体一臂的顶端,第Ⅳ类为较弱的45S rDNA位于染色体一臂内部。八倍体柳枝稷不同栽培品种以及同一栽培品种不同个体间45S rDNA信号复杂性的成因可能与染色体同源重组以及染色体结构变异密切相关。     

国产十种乌头的染色体研究

本文描述了我国产毛茛科乌头属的10个种的染色体数目和形态。 根据该属染色体基 数为8(x＝8),可将这些种归为二倍体、四倍体、六倍体和八倍体种。 10种乌头的染色体在数目、大小、结构上表现出相关性。其中具根状茎而多年生的种多为二倍体,染色体形态较大,第3-7号多为近端着丝点(st)染色体, 具块茎而二年生的种多为多倍体,染色体形态较小,第3-7号多为近中着丝点(sm)染色体。染色体演化方向可能是二倍体→多倍体、大→ 小、st→sm。这进一步证明根状茎种较块茎种原始。根据染色体数目和形态,又可将这些种的染色体分为两类。这进一步支持了目前分类学上的牛扁亚属和乌头亚属的划分。本文还讨论了这些种的有关分类问题。     

金沙江中游地区红山茶组植物的Giemsa C－带研究

研究了金沙江中游地区红山茶组植物的ＧｉｅｍｓａＣ－带。该地区的红山茶植物以四倍体为主,个别居群为二倍体或六倍体。居群间的Ｃ－带差异明显,Ｃ－带多出现在染色体端部。在四倍体和六倍体的Ｃ－带带型中,只能找到２条显相同Ｃ－带的同源染色体,通过与其它地区的红山茶植物进行比较,发现红山茶组植物的倍性从华东,华南经贵州,四川向云南逐渐增高,显Ｃ－带的染色体与染色体总数之比随信性增加而减少。文中指出华东或华南可能是红山茶组植物的起源地,而金沙江中游地区是其现代分化中心,这一地区红山茶的多倍体类群可能是异源起源的。     

Screening and identification of female-specific DNA sequences in octaploid sturgeon using comparative genomics with high-throughput sequencing

In this study, six candidate female-specific DNA sequences of octaploid Amur sturgeon (Acipenser schrenckii) were identified using comparative genomic approaches with high-throughput sequencing data. Their specificity was confirmed by traditional PCR. Two of these sex-specific sequences were also validated as female-specific in other eight sturgeon species and two hybrid sturgeons. The identified female-specific DNA fragments suggest that the family Acipenseridae has a ZZ/ZW sex-determining system. However, one of the two DNA sequences has been deleted in some sturgeons such as Sterlet sturgeon (Acipenser ruthenus), Beluga (Huso huso) and Kaluga (H. dauricus). The difference of sex-specific sequences among sturgeons indicates that there are different sex-specific regions among species of sturgeon. This study not only provided the sex-specific DNA sequences for management, conservation and studies of sex-determination mechanisms in sturgeons, but also confirmed the capability of the workflow to identify sex-specific DNA sequences in the polyploid species with complex genomes.     

Development of new microsatellite primers for green and white sturgeon

Sixty-eight primer sets for microsatellite loci were developed from microsatellite motif enriched genomic libraries of pooled DNA from the polyploid green and white sturgeon (Acipenser medirostris and A. transmontanus). Four individuals from each species were screened for polymorphism at these loci. Forty-eight loci amplified in both species, and some exhibited species-specific amplification for white or green sturgeon (8 and 12 loci, respectively). The number of alleles per locus ranged from one to 12. At least 68% of the green and 65% of the white sturgeon loci we developed are polysomic.     

Evidence for positive selection of taurine genes within a QTL region on chromosome X associated with testicular size in Australian Brahman cattle

Background

Evolution of sturgeons and paddlefishes (order Acipenseriformes) is inherently connected with polyploidization events which resulted in differentiation of ploidy levels and chromosome numbers of present acipenseriform species. Moreover, allopolyploidization as well as autopolyploidization seems to be an ongoing process in these fishes and individuals with abnormal ploidy levels were occasionally observed within sturgeon populations. Here, we reported occurrence of Siberian sturgeon (Acipenser baerii) male with abnormal ploidy level for this species, accessed its ploidy level and chromosome number and investigate its potential sterility or fertility in comparison with normal individuals of sterlet (A. ruthenus), Russian sturgeon (A. gueldenstaedtii) and Siberian sturgeon (A. baerii).

Results

Acipenser ruthenus possessed 120 chromosomes, exhibiting recent diploidy (2n), A. gueldenstaedtii and A. baerii had ~245 chromosomes representing recent tetraploidy (4n), and A. baerii male with abnormal ploidy level had?~?368 chromosomes, indicating recent hexaploidy (6n). Genealogy assessed from the mtDNA control region did not reveal genome markers of other sturgeon species and this individual was supposed to originate from spontaneous 1.5 fold increment in number of chromosome sets with respect to the number most frequently found in nature for this species. Following hormone stimulation, the spontaneous hexaploid male produced normal sperm with ability for fertilization. Fertilization of A. baerii and A. gueldenstaedtii ova from normal 4n level females with sperm of the hexaploid male produced viable, non-malformed pentaploid (5n) progeny with a ploidy level intermediate to those of the parents.

Conclusion

This study firstly described occurrence of hexaploid individual of A. baerii and confirmed its autopolyploid origin. In addition to that, the first detailed evidence about fertility of spontaneous hexaploid sturgeon was provided. If 1.5 fold increment in number of chromosome sets occurring in diploids, resulted triploids possess odd number of chromosome sets causing their sterility or subfertility due to interference of gametogenesis. In contrast, 1.5 fold increment in number of chromosome sets in naturally tetraploid A. baerii resulted in even number of chromosome sets and therefore in fertility of the hexaploid specimen under study.     

Genome duplication events and functional reduction of ploidy levels in sturgeon (Acipenser, Huso and Scaphirhynchus).

Sturgeon (order Acipenserformes) provide an ideal taxonomic context for examination of genome duplication events. Multiple levels of ploidy exist among these fish. In a novel microsatellite approach, data from 962 fish from 20 sturgeon species were used for analysis of ploidy in sturgeon. Allele numbers in a sample of individuals were assessed at six microsatellite loci. Species with approximately 120 chromosomes are classified as functional diploid species, species with approximately 250 chromosomes as functional tetraploid species, and with approximately 500 chromosomes as functional octaploids. A molecular phylogeny of the sturgeon was determined on the basis of sequences of the entire mitochondrial cytochrome b gene. By mapping the estimated levels of ploidy on this proposed phylogeny we demonstrate that (I) polyploidization events independently occurred in the acipenseriform radiation; (II) the process of functional genome reduction is nearly finished in species with approximately 120 chromosomes and more active in species with approximately 250 chromosomes and approximately 500 chromosomes; and (III) species with approximately 250 and approximately 500 chromosomes arose more recently than those with approximately 120 chromosomes. These results suggest that gene silencing, chromosomal rearrangements, and transposition events played an important role in the acipenseriform genome formation. Furthermore, this phylogeny is broadly consistent with previous hypotheses but reveals a highly supported oceanic (Atlantic-Pacific) subdivision within the Acipenser/Huso complex.     

The use of microsatellite loci for identification of sturgeon species (Acipenseridae) and hybrid forms

The genetic polymorphism of ten sturgeon species that inhabit the territory of the Russian Federation (Russian sturgeon, Siberian sturgeon, Amur sturgeon, Sakhalin sturgeon, Persian sturgeon, ship sturgeon, sterlet, stellate sturgeon, beluga, and kaluga) was examined at five microsatellite loci (Afug41, Afug51, An20, AoxD161, AoxD165) (in total, 3821 individuals). The examined loci were successfully amplified with the same primer set in all species tested and demonstrated a high level of variation. Alleles specific to different species have been identified, which allows them to be used to identify species of sturgeon and their food products. In addition, the possibility of identifying hybrid forms was demonstrated. The assignment test performed in the STRUCTURE software program showed a high probability of correctly assigning each individual to its species based on genotyping with five microsatellite loci examined (96–98%, on average). However, for Russian and Persian sturgeon, the rate of proper species assignment was considerably lower (75 and 84%, respectively).

     

Polyploidy‐associated genomic instability in Arabidopsis thaliana

Formation of polyploid organisms by fertilization of unreduced gametes in meiotic mutants is believed to be a common phenomenon in species evolution. However, not well understood is how species in nature generally exist as haploid and diploid organisms in a long evolutionary time while polyploidization must have repeatedly occurred via meiotic mutations. Here, we show that the ploidy increased for two consecutive generations due to unreduced but viable gametes in the Arabidopsis cyclin a1;2‐2 (also named tardy asynchronous meiosis‐2) mutant, but the resultant octaploid plants produced progeny of either the same or reduced ploidy via genomic reductions during meiosis and pollen mitosis. Ploidy reductions through sexual reproduction were also observed in independently generated artificial octaploid and hexaploid Arabidopsis plants. These results demonstrate that octaploid is likely the maximal ploidy produced through sexual reproduction in Arabidopsis. The polyploidy‐associated genomic instability may be a general phenomenon that constrains ploidy levels in species evolution. genesis 48:254–263, 2010. © 2010 Wiley‐Liss, Inc.     

Development and standardization of disomic microsatellite markers for lake sturgeon genetic studies

Lake sturgeon (Acipenser fulvescens) are of conservation concern in North America. To facilitate the recovery of this fish species, an understanding of their population genetic structure is necessary to develop and implement spatially and temporally appropriate management actions. Until recently, few genetic data using nuclear loci have been collected, primarily due to the paucity of suitable genetic markers because most microsatellite loci in lake sturgeon appeared to be tetrasomic. The authors identified nine microsatellite loci (from 254 examined) that were putative polymorphic disomic loci and tested their conformance to a disomic mode of inheritance using three lake sturgeon families. The objectives of the study were to: (i) confirm the disomic status of the nine loci through inheritance testing, and (ii) standardize the genetic markers among participating laboratories. At all nine loci, disomic inheritance were confirmed, and all nine loci segregated independently in the 26 of 36 loci pairs possible to test. One of the nine loci showed non‐Mendelian segregation, possibly due to meiotic drive and/or selection. Three progeny had peak patterns inconsistent with disomy at one or more loci. The nine loci when combined with four microsatellite loci previously confirmed in other studies as disomic in lake sturgeon now yield a suite of 13 microsatellite markers. These 13 markers have been standardized among four other laboratories to facilitate building an inter‐laboratory genetic database for lake sturgeon.     

Isolation and characterization of thirteen microsatellite loci in four diploid species of Ranunculus cantonensis polyploid complex

Ranunculus is a cosmopolitan genus, and the evolution of the genus has been influenced by both polyploidy and hybridization. The interactions between some species of Ranunculus are regarded as a natural model system for studying hybridization. Four diploid species of the R. cantoniensis polyploid complex have relatively little reproductive isolation from each other, and they form a complicated hybrid pool in a multispecies hybrid zone. To clarify the genetic structure and spatiotemporal dynamics of gene flow in this multispecies hybrid zone, 13 polymorphic microsatellite loci were isolated from R. silerifolius var. dolicanthus. The loci's characteristics were observed in 30 individuals. The number of alleles per locus ranged from 6 to 20, and the PIC values ranged from 0.350 to 0.914. Eleven loci were significantly biased from the Hardy–Weinberg equilibrium. The 13 microsatellite loci were shown to be successfully amplified in three other related species. These polymorphic microsatellites will be powerful molecular tools for further studies of the Ranunculus genetic structure and gene flow.     

湖鲟微卫星DNA引物应用于中华鲟亲子关系分析的初步研究

利用湖鲟(Acipenser fulvescens)的4对微卫星引物对中华鲟随机个体样本进行PCR扩增,分析电泳结果发现,4对引物均可在中华鲟个体中得到稳定的同源序列,其中2对引物所探测到的等位基因数目较多,在个体间表现出较高的多态性,利用它们产生的DNA指纹图谱,能够对1999年度获得的中华鲟亲鱼样本进行有效的个体区分。并且这两对湖鲟的微卫星引物在对1999年度已知亲本的同一家系中的中华鲟随机个体的分析中,表现为按照孟德尔方式进行共显性遗传。证明这2对微卫星引物可以用于鉴别中华鲟人工放流个体和自然繁殖个体。     

跨种扩增筛选大鳄龟微卫星标记及遗传多样性分析

跨种扩增是一种能够快速、有效地获得物种微卫星标记的方法。本研究利用在近缘种中已发表的微卫星DNA引物,对大鳄龟（Macroclemys temminckii）进行跨种PCR扩增,在合成的69对引物中获得8对具有多态性的微卫星位点。对PCR扩增产物进行统计,得出观测杂合度（H_o）的范围是0.041 7~0.954 5,平均为0.384 8;期望杂合度（H_E）的范围为0.041 7~0.811 8,平均为0.479 1;多态信息含量范围为0.040 0~0.759 2,平均为0.423 2;经过卡方检验后,部分微卫星位点符合哈迪-温伯格平衡。总体来说,这些位点是研究大鳄龟遗传结构的良好分子标记。