Genetic mapping of a new flowering time gene on chromosome 3B of wheat

The single chromosome substitution lines of chromosome 3B of the Czech alternative wheat variety Česká Přesívka (CP 3B) into two spring varieties Zlatka and Sandra, revealed clear differences in flowering time compared to the recipient varieties. To map this gene(s), recombinant substitution lines for chromosome 3B were produced from crosses of the substitution lines with their recipient parents and genetic maps developed using SSR markers. Two populations were mapped, Sandra//Sandra 3B/Sandra (CP 3B) and Zlatka//Zlatka/Zlatka (CP 3B). Combining the genotype data with phenotype data on flowering time in five independent experiments under natural long day or controlled short day conditions revealed a single flowering time QTL. This gene had an additive effect of 1–6 days, depending on environment and genetic background, and was mapped in both populations to a position in the region of marker Xbarc164 near the centromere on the long arm of 3B. Comparisons of the genetic maps with other 3B maps developed by the authors indicated that the QTL may be homologous to a QTL segregating in UK germplasm.     

开发实用的染色体加倍体系构建成烟草DH群体

通过对4种染色体加倍方法(烟草浸花药法、浸花培苗法,叶片再生法及浸腋芽法等)的比较研究表明：以4g／L浓度的秋水仙碱浸泡花药法加倍率最高达47．5％～75％,其次为叶片再生法(30％～36．67％)和浸苗法(16．67％～32．35％),而浸腋芽法较低(17．78％)。前两种方法加倍率虽高,但有较高畸形苗比率,叶片再生法工序较繁琐。作者认为烟草加倍单倍体产生,应以采用浸苗法为主。在使用秋水仙碱浸苗加倍时,添加DMSO可明显促进秋水仙碱的加倍效率,且促进作用随时间延长而提高：从高效、快捷和节约等原则考虑,我们开发了烟草有效的染色体加倍体系,以4g／L的秋水仙碱 20g／LDMSO溶液浸苗48h的效果最好。对两个组合烟草单倍体苗浸12h以上,其染色体加倍效率达到对照的2．30-2．93倍。本试验用较低浓度秋水仙碱添加DMSO有利于节约实验费用。通过完善染色体加倍技术程序已构建成2个DH群体,供基因定位研究。     

大西洋鲑的染色体核型

采用体内注射小牛血清、PHA培养法，取鱼体肾细胞经空气干燥法制备大西洋鲑（Salmo salar)的染色体玻片，并进行核型分析。试验结果表明：大西洋鲑的染色体数目为2n=56，其核型为18m＋38t，臂数NF：74。     

染色体单片段代换系的构建及应用于QTL精细定位

染色体片段代换系(Chromosome Segment Substitution Lines,CSSLs),又称为代换系(Substitution lines,SLs)或导入系(Introgression lines,ILs)。只含一个代换片段的代换系,即单片段代换系(Single Segment Substitution Lines,SSSLs)是理想的代换系。单片段代换系只有代换片段与受体亲本不同,其它遗传背景与受体亲本完全一致,对代换区段中的QTL进行分析时遗传背景干扰很小,有利于QTL的分析,不少学者利用单片段代换系材料对许多QTL进行了鉴定和精细定位,并克隆了一些重要性状的QTL。本文介绍了染色体单片段代换系构建的原理和利用微卫星标记(SSR）进行单片段代换系鉴定的方法,讨论了代换系构建过程中值得注意的问题,并对用染色体单片段代换系进行QTL精细定位做了展望。     

小麦全基因组NBS类R基因分析及2AL染色体NBS-SSR特异标记开发

NBS (nucleotide binding site)类基因是植物界中最重要的一类抗病基因。用信息学方法从普通小麦(Triticum aestivum L.)全基因组中分离出2406条含有NBS结构的完整蛋白序列,每条包含48~2272个氨基酸。根据NBS结构域两端是否连接CC或LRR结构域,将TaNBS分为N、CN、NL和CNL4类。对TaNBS所在scaffold序列的SSR位点进行诊断,从1203条scaffold序列上发现2177个SSR位点,以二碱基重复位点最多,占73.5%。针对小麦2AL染色体上的51个SSR位点开发标记,缺体-四体和双端体验证结果表明,有39个标记(76.5%)为2AL特异标记,其中24个特异标记在抗白粉病材料Khapli (2AL上携带Pm4a)和感病材料Chancellor间存在多态性。利用近等基因系Khapli/8*Cc筛选出3个可能与Pm4a连锁的NBS-SSR标记,分别是Sxaas_2AL22、Sxaas_2AL39和Sxaas_2AL46。本研究开发的与抗病序列紧密连锁的特异SSR标记可用于2AL染色体上抗病新基因的检测以及已有抗病基因的候选序列筛选。     

Genetic diversity of the Mexican Lidia bovine breed and its divergence from the Spanish population

Lidia bovine breed exists since the XIV century in the Iberian Peninsula. These animals were initially produced for meat but some, showing an aggressive behaviour which difficulted their management, were used to participate in popular traditional and social events. A specialization of the breed giving rise to the original Lidia population is documented in Spain since mid-XVIII century. Following the same tradition than in the Spanish population, Mexico used aggressive animals at the beginning of the XX century until two families of breeders started importing Lidia breed bovines from Spain with the aim of specializing their production. Each family (Llaguno and González) followed different breeding managements, and currently, most of the Lidia Mexican population derives from the Llaguno line. Although genetic structure and diversity of the Spanish population have been studied (using autosomal microsatellite markers, Y chromosome DNA markers and mitochondrial DNA sequences), the Mexican population is not analysed. The aim of the study was to assess both the genetic structure and diversity of the Mexican Lidia breed and its relationship with the original Spanish population using the same molecular tools. A total of 306 animals belonging to 20 breeders issued from both existing Mexican families were genotyped, and the genetic information was compared to the previously existing Spanish information. Slightly higher levels of genetic diversity in Mexican population were found when comparing to the Spanish population, and the variability among populations accounted for differences within them showing mean values of 0.18 and 0.12, respectively. Animals from the Mexican breeders, belonging to each of the two families, clustered together, and there was little evidence of admixture with the Spanish population. The analysis of Y chromosome diversity showed a high frequency of the H6 haplotype in the Mexican population, whereas this haplotype is rare in the Spanish, which is only found in the Miura (100%) and Casta Navarra (38%) lineages. Mitochondrial DNA revealed similar haplotypic pattern in both Spanish and Mexican populations, which is in accordance with most of the Mediterranean bovine breeds. In conclusion, as the Mexican Lidia population had initially a small number of founders and its current population has been reared isolated from their Spanish ancestors since a long time, these bottleneck effects and a combination of mixed cattle origin are the factors that might erase any trace of the Spanish origin of this population.     

双壳贝类染色体标本制备技术的优化

本研究以牡蛎、咬齿牡蛎鳃细胞为材料,采用PHA处理法、低温同步化法及活体去壳法进行预处理,制作成染色体标本;以马氏珠母贝、企鹅珍珠贝担轮幼虫为材料,采用PHA促繁殖获取胚胎法,制作胚胎染色体标本。对不同时间不同处理方式获得牡蛎染色体分裂相比例、PHA促繁殖获取胚胎法获得马氏珠母贝染色体分裂相比例进行统计。结果发现,PHA处理法在24h时能获取最多分裂相（3．50‰）,低温同步化法在72h时能获得最多分裂相（2．20‰）,活体去壳法始终保持较高分裂相比例（3．60‰）,PHA促繁殖获取胚胎法获得最大分裂相比例（10．00‰）。4种方法都能获得理想的中期分裂相数目。其中,低温同步化法缺点是温度难控制、预处理时间长;活体去壳法易导致贝类死亡;PHA促受精获取胚胎法缺点是胚胎的获取受繁殖季节限制。PHA处理法预处理时间短、获取分裂相数目多,无疑是贝类成体染色体制备的最好的方法。同时,PHA也为贝类人工繁殖提供了一种更为有效的手段。     

蓝粒小麦4Ag染色体的显微分离与鉴定

本研究采用显微分离技术从蓝粒小麦(Triticum aestivum L.(2n=6x=42)×Thinopyrum ponticum Liu &Wang(2n=10x=70))根尖细胞中期分裂相中分离出具有4Ag染色体形态特征的单条染色体,对分离后的细胞进行原位杂交(FISH),结果显示细胞中所剩的和显微分离到的单条染色体均为4Ag染色体.利用Sau3A接头介导的PCR方法对分离出的单条4Ag染色体进行体外扩增,扩增的DNA片段大小约为200～2 000 bp,主要集中在250～750 bp之间.以DIG-dUTP标记的蓝粒基因组DNA为探针,与该扩增产物进行Southern杂交,结果表明显微分离出的染色体得到了有效的扩增.利用该分离染色体的PCR扩增产物为探针,对蓝粒小麦进行原位杂交,证明显微分离的染色体体外扩增片段确实来源于4Ag染色体.本研究拓宽了染色体显微分离的范围,为构建4Ag染色体文库和克隆位于该染色体上的重要农艺性状基因提供了新途径.     

人工合成甘蓝型油菜减数分裂中的染色体行为观察

以白菜(Brassica rapaL.,2n=20,AA)为母本,芥蓝(B.oleraceaL.,2n=18,CC)为父本,通过种间杂交、子房培养和染色体加倍,获得新合成甘蓝型油菜(B.napusL.,2n=38,AACC)株系35个。细胞学观察结果表明,新合成甘蓝型油菜的花粉母细胞(PMCs)内染色体在早双线期聚集成两个染色体群,具有较为明显的界限;随后逐渐形成单个二价体,直至终变期产生全部19个二价体,或一些四价体和二价体;在个别PMCs内19个二价体呈9∶10的分开排列;在后期Ⅰ/末期Ⅰ以及第二次减数分裂过程中,染色体分离较为正常。在讨论中提出甘蓝型油菜的染色体在长期的进化过程中所产生的结构变异可能主要来自于A、C基因组内部染色体间交换的假说。