亚麻外源DNA导入的技术与实践

为了尽快选育亚麻新品种,我所从20世纪90年代对亚麻外源DNA导入技术研究,从DNA提取、子房内部结构、外源DNA导入时间、方法、后代的变异、品种选育的应用等进行了一系列的研究,并取得了一定的成就,选育出等黑亚14、16、17等3个亚麻品种及一批优良品系。利用该技术进行亚麻育种,可缩短育种年限,加速育种进程。     

Chloroplast DNA molecular characterization and leaf volatiles analysis of mint (Mentha; Lamiaceae) populations in China

Non-coding chloroplast DNA (cpDNA) was used to evaluate genetic diversity and relationships within the section Mentha collected from China. Leaf volatiles of Mentha spicata accessions and its parent were analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). To this end, genetic diversity of 30 Mentha accessions from different geographic origins of China, representing six species and two hybrids, was assessed. Molecular studies grouped the samples mainly according to genetic relationship established by conventional methods. Cluster analysis based on the leaf volatiles chemical composition of M. spicata accessions defined two main chemotypes: 1,8-cineole-piperitenone oxide type and limonene-carvone type. The results suggested that there was a high genetic variability among individuals of Mentha in China. A distinct correlation between cpDNA marker and volatile oils in M. spicata accessions was revealed. Moreover, the chemical composition followed maternal inheritance in M. spicata accessions. In conclusion, the genetic diversity of Mentha populations as shown in this study should play a critical role in future selection and breeding programs.     

The implications for aquaculture practice of epigenomic programming of components of the endocrine system of teleostean embryos: lessons learned from mammalian studies

Epigenetics is the study of changes in gene expression patterns that occur without any modification of the underlying nucleotide sequence of the DNA. Modifications of the so‐called epigenome include complex transient or permanent chemical changes of the DNA or histone proteins resulting in the suppression or enhancement of gene expression, together with an array of post‐translational events that modify the translational products. Epigenomic programming (EP) of the genome is an essential component of embryonic development in animals from the totipotent fertilized egg to the pluripotent stem cells, stem cell differentiation and final tissue and organ formation. Many of these EP processes are influenced transiently and some permanently by environmental influences. In eutherian mammals, environmentally related EP of embryos is linked to permanent changes in the phenotype of the progeny, some of which have been associated with adult onset metabolic disorders. Moreover, because some of the epigenetic remodelling occurs in both the soma and germ line, the resultant phenotypic characteristics (some of which are linked to disease states) may be heritable. Although far less is known about the effects of environmentally linked EP on the ontogeny of fishes, the available information suggests that the EP processes are similar amongst all vertebrates, and there are clear parallels between fish and mammals that are discussed in this paper. Our perspective takes the well‐established findings in mammals and uses them to proactively extrapolate to the as yet under‐recognized implications of EP for fish biology and for fish production in intensive aquaculture.     

哈克尼西棉细胞质陆地棉雄性不育系orf160的克隆及遗传转化

以哈克尼西棉细胞质陆地棉雄性不育系及其保持系、恢复系为材料,克隆并测序线粒体功能基因的侧翼序列。通过生物信息学分析,在atp4下游发现一个哈克尼西棉细胞质不育系特有的orf160。orf160全长480 bp,N端与atp6序列部分同源,C端序列与核序列部分同源;编码的159个氨基酸序列,与膜蛋白、细胞周期蛋白具有部分同源性。以含有线粒体导肽的GFP瞬时表达载体转化后,在洋葱表皮的细胞膜上观察到绿色荧光,表明线粒体定向载体正常表达。构建酵母和植物表达载体,分别转化酵母、烟草和拟南芥;同对照相比,转基因酵母菌斑畸形且生长缓慢;转基因拟南芥和烟草种子大部分畸形,T₁和T₂代植株的结实率和种子发芽率降低,花粉着色比野生型浅。结果表明,orf160基因的表达影响受体正常发育。     

3种椰子基因组DNA提取方法的比较

旨在得到一套快速提取椰子DNA的方法。采用CTAB小样法、SDS-CTAB法以及PVP法提取椰子组织的DNA,并通过后续的PCR扩增和酶切等实验比较了3种方法所提取的DNA质量的优劣。将CTAB小样法提取的DNA与SDS-CTAB法和PVP法提取的DNA进行了比较,CTAB小样法提取的DNA的OD260/OD280(1.839)较另外2种方法SDS-CTAB法(1.709)和PVP法(1.613)的值更高,CTAB小样法提取的DNA适合于PCR扩增,扩增的条带清晰,且特异性好。此外,CTAB小样法提取的DNA,酶切也较为均匀。CTAB小样法能高效地提取椰子的DNA,并且能一次提取大量的高质量DNA样品,因而能满足高通量的实验要求,同提取DNA能较好地满足下游的分子生物学研究。     

濒危树种金钱松RAPD体系的建立和遗传多样性分析

通过优化随机扩增多态性DNA（RAPD） 反应体系,对浙江省不同来源的金钱松Pseudolarix amabilis进行遗传多样性分析。结果表明：18条随机引物在62个样品中可检测到172个可重复位点,其中多态性位点有170个,多态性比率为99.2%。聚类分析的结果表明：同一来源金钱松材料聚为一类,说明金钱松天然群体的RAPD多态性分类和其地理分布有一定的关系,但也有特殊个体不一致,表明金钱松天然种群体存在较高的遗传多样性,并建议对金钱松进行就地保存和迁地保存。图2表3参31     

雷公藤总DNA提取方法的研究

由于雷公藤叶片富含多糖及酚类物质,要获得高质量的DNA有一定难度。本研究以雷公藤植物叶片为材料,对传统提取方法进行改进、优化,比较了4种不同的提取方法,最终获得了一种以CTAB法为基础,试剂盒纯化的获得高质量DNA的方法。试验结果表明,使用该方法从雷公藤叶片中分离的DNA,纯度高、杂质少,且相对于其他方法,试验成功率明显提高,适用性强。     

玉米种子纯度鉴定中DNA提取方法的比较研究

分别以玉米干种子胚、幼芽和幼苗为材料提取DNA,并对所得DNA的纯度和质量进行检测,结果显示,利用玉米干种子胚提取的DNA质量及分子量大小与幼苗法相当,完全可以满足SSR分子标记对玉米杂交种纯度的鉴定。     

SDS-CTAB法提取烟草病圃土壤微生物总DNA

采用SDS-CTAB法提取了烟草病圃土壤中微生物的总DNA,经过纯化后获得的DNA大小约为21 kb,OD260/OD280高于1.70;用细菌和真菌保守序列引物进行PCR扩增,琼脂糖凝胶电泳能检测到预期大小的扩增条带。     

蝴蝶兰叶绿体DNA微卫星分析与标记开发

为开发兰科植物叶绿体微卫星(cpSSR)引物,对台湾蝴蝶兰叶绿体全序列进行cpSSR统计分析。结果表明台湾蝴蝶兰SSR以单碱基重复为主,占总数的78.3%。在单碱基重复中又以A和T重复为主,占95.8%。重复方式以完全重复为主,占总数的70%。开发出3对cpSSR引物。