Physical mapping of rDNA in Dendranthema nankingense and its close related species by fluorescent in situ hybridization

Distribution of 18S-26S rDNA on the chromosomes of Dendranthema nankingense and its two close-related species D. nankingense and D. lavendulifolium was studied by fluorescent in situ hybridization with a JHD 2-15A DNA clone. Six rDNA loci were found in D. nankingense, eight in D. lavendulifolium, and twelve in D. indicum. The rDNA loci number and pattern were applied to analyze the phylogenetic relationship of the three closely related species.     

菊花CmDFR与CmANS基因启动子序列克隆与瞬时表达分析

DFR和ANS是花青素合成途径下游两个关键的结构基因,其不仅决定花青素苷最终结构及其呈色,还在花器官中特异性表达,研究其启动子区域对花色改良的分子育种具有重要参考价值.利用接头染色体步移法( genome walking),从菊花的叶片基因组DNA中克隆到了2个DFR基因同源序列CmDFR的启动子片段,1个ANS基因同源序列CmANS的启动子片段.生物信息学软件分析结果显示,这3个启动子片段除了含有多个TATA-box、CAAT-box等基本的启动子元件,还含有很多与MYB转录因子相结合的元件,以及G-box等参与光响应的顺式作用元件.利用双酶切方法,将克隆到的启动子片段替换栽体pB1121上的35S启动子,将新构建的植物表达栽体转入农杆菌中,采用叶盘法侵染菊花叶片和花瓣进行瞬时表达研究.结果表明,这3个启动子片段均具有驱动下游报告基因表达的功能.因此,可以开发成菊花分子育种的有效基因构件.     

A molecular genetic linkage map of mouse chromosome 13 anchored by the beige (bg) and satin (sa) loci

A molecular genetic linkage map of mouse chromosome 13 was constructed using cloned DNA markers and interspecific backcross mice from two independent crosses. The map locations of Ctla-3, Dhfr, Fim-1, 4/12, Hexb, Hilda, Inhba, Lamb-1.13, Ral, Rrm2-ps3, and Tcrg were determined with respect to the beige (bg) and satin (sa) loci. The map locations of these genes confirm and extend regions of homology between mouse chromosome 13 and human chromosomes 5 and 7, and identify a region of homology between mouse chromosome 13 and human chromosome 6. The molecular genetic linkage map of chromosome 13 provides a framework for establishing linkage relationships between cloned DNA markers and known mouse mutations and for identifying homologous genes in mice and humans that may be involved in disease processes.     

玉米Lc基因植物表达载体构建及菊花转化

Lc基因是从玉米中分离得到的与花青素合成相关的调节基因,它在多种植物中的异源表达可以增加花青素的含量.本研究对pBI121载体Gus基因后的终止子进行2次PCR扩增,在原Sac Ⅰ酶切位点后添加了新的Sac Ⅰ酶切位点,利用组织化学染色法检测,结果表明改造后的载体上的Gus基因能正常表达,终止子功能正常,载体改造成功.用改造的pBI121N构建了含有Lc基因的植物表达载体pBl121N-Lc,利用农杆菌介导法转化菊花叶盘,获得了19株生根抗性苗.通过提取抗性苗基因组总DNA,PCR扩增Lc基因和CaMv35S启动子获得了11个阳性株系,PCR结果表明Lc基因已经转入菊花中.同时,在已获得的转基因植株中发现7个株系的根系有变红的现象.     

Application of Genomic SSR Locus Polymorphisms on the Identification and Classification of Chrysanthemum Cultivars in China

The Chinese traditional chrysanthemum is a notable group of chrysanthemums (Chrysanthemum×morifolium Ramat.) in which the phenotypic characteristics richly vary. At present, there is a serious controversy regarding homonyms and synonyms within this group. Moreover, the current international chrysanthemum classification systems are not comprehensive enough to be used on Chinese traditional chrysanthemums. Thus, we first identified a broad collection of 480 Chinese traditional chrysanthemum cultivars using the unique DNA fingerprints and molecular identities that were established by 20 simple sequence repeat markers. Five loci, which distinguished all of the selected cultivars, were identified as the core loci to establish unique fingerprints and molecular identities with 19 denary digits for each cultivar. A cluster analysis based on Nei''s genetic distance indicated that the selected cultivars were clustered according to their horticultural classification. Population structure analysis was subsequently performed with K values ranging from 2 to 14, and the most likely estimate for the population structure was ten subpopulations, which was nearly consistent with the clustering result. Principal component analysis was further performed to verify the classification results. On the basis of the Q-matrices of K = 10, a total of 19 traits were found to be associated with 42 markers. Taken together, these results can serve as starting points for the identification and classification of chrysanthemums based on the polymorphism of microsatellite markers, which is beneficial to promote the marker-assisted breeding and international communication of this marvelous crop.     

植物花青素苷转运机制的研究进展

花青素苷的合成过程是生物学上研究得较为清楚的代谢通路之一,但其最后阶段的分子机制即花青素苷从细胞质被转运至中央液泡的过程却仍不清晰。最近研究者们刚刚开始对类黄酮化合物的转运过程进行动态的描绘,迄今共提出了4种花青素苷转运模型,发现了4类与花青素苷转运过程相关的转运蛋白:谷胱甘肽转移酶、多药耐药抗性相关蛋白、多药和有毒化合物排出家族和同源于哺乳动物的胆红素易位酶同族体,并对这4种转运体及相关基因的功能进行了初步研究。尽管已经提出了不同的花青素苷转运模型,但仍然缺乏对不同物种不同类型花青素苷向液泡转运及在液泡中沉积的细胞学和亚细胞学研究。根据获得的信息,可以通过开展基因序列分析、基因表达分析、亚细胞定位和互补试验等,探求转运蛋白的功能及其作用位置,更好地解析植物体内花青素苷的转运机制。     

38个中国大菊品种染色体核型参数的统计分析

为了探讨中国传统大菊（Chrysanthemum×morifolium Ramat.）品种间的遗传差异,选取了38个品种进行核型参数和染色体数目观察和统计。结果表明：多数大菊品种的染色体数目介于50–60之间,为六倍体基础上的非整倍体;其中仅有2个品种染色体数目少于50,3个品种多于60;黄香梨（C.×morifolium‘Huangxiangli’）是唯一染色体数目达到73的品种,推测其为八倍体。观察品种多数为较对称的核型（2A或2B型）;具有随体染色体的品种有30个,占观察品种总数的79%,且古老品种的随体染色体数目较多,千手观音（C.×morifolium‘Qianshouguanyin’）的中期分裂相里最多可观察到6个随体;臂比均值（MAR）部分集中在1.5–2.0之间,臂比方差（VAR）则集中在0.1–0.5之间,臂比均值与臂比方差呈显著正相关,其相关系数（r2）为0.91。各瓣型之间的臂比均值差别不大;古老品种与现代品种在臂比均值和臂比方差上并未表现出明显的差异。三大瓣型（平瓣、匙瓣和管瓣）的核型不对称系数（As.K.C.）差异不显著,其在古老品种中较为一致,而现代品种之间的差异较大。这些数据表明,核型参数对菊花的品种鉴定、分类和遗传分析具有重要参考价值。