基因工程对改良观赏植物品质的研究进展

基因工程技术已在观赏植物的花色、花形、花香、花期、抗性等方面大量应用并取得了较大的进展,为花卉的性状和品质改良提供了全新的思路和手段。作者就近年来基因工程技术在观赏植物品质研究方面进行了综述,并对观赏植物基因工程存在的问题进行了探讨。     

大红甜橙遗传转化高效再生体系的建立

以大红甜橙的实生苗为试材,研究6-BA与IAA的不同浓度组合和Km不同浓度对大红甜橙不同外植体离体再生的影响和IBA的不同浓度对不定芽生根的影响,建立了高效的大红甜橙遗传转化再生体系.该体系的最佳条件是:1)以实生苗上胚轴为外植体.2)以MS无机盐 100mg/L肌醇 0.2mg/LV-B1 1mg/LV-B6 1mg/L烟酸 3%蔗糖 0.8%琼脂(pH5.8) 3mg/L6-BA为不定芽诱导培养基,在26℃黑暗条件下培养7d后,转移至光/暗周期16/8h的条件下诱导不定芽的分化(40d分化率达90%).以1/2MS 3%蔗糖 0.7%琼脂 2.0mg/LIBA为不定芽的生根培养基(40d生根率达94.7%).3)筛选遗传转化转化体的Km质量浓度为50mg/L.     

Multilocus sequence typing detects new Piscirickettsia salmonis hybrid genogroup in Chilean fish farms: Evidence for genetic diversity and population structure

Piscirickettsia salmonisis the causative bacterial pathogen of piscirickettsiosis, a salmonid disease that causes notable mortalities in the worldwide aquaculture industry. Published research describes the phenotypic traits, virulence factors, pathogenicity and antibiotic‐resistance potential for various P. salmonisstrains. However, evolutionary and genetic information is scarce for P. salmonis. The present study used multilocus sequence typing (MLST) to gain insight into the population structure and evolution of P. salmonis. Forty‐two Chilean P. salmonisisolates, as well as the type strain LF‐89^T, were recovered from diseased Salmo salar, Oncorhynchus kisutchand Oncorhynchus mykissfrom two Chilean Regions. MLST assessed the loci sequences of dnaK, efp, fumC, glyA, murG, rpoD and trpB. Bioinformatics analyses established the genetic diversity among P. salmonis isolates (H = 0.5810). A total of 23 sequence types (ST) were identified, 53.48% of which were represented by ST1, ST5 and ST2. Population structure analysis through polymorphism patterns showed few polymorphic sites (218 nucleotides from 4,010 bp), while dN/dS ratio analysis indicated purifying selection for dnaK, epf, fumC, murG, and rpoD but neutral selection for the trpB loci. The standardized index of association indicated strong linkage disequilibrium, suggesting clonal population structure. However, recombination events were detected in a group of seven isolates. Findings included genogroups homologous to the LF‐89^T and EM‐90 strains, as well as a seven‐isolate hybrid genogroup recovered from both assessed regions (three O. mykiss and four S. salar isolates). The presented MLST scheme has comparative potential, with promising applications in studying distinct P. salmonis isolates (e.g., from different hosts, farms, geographical areas) and in understanding the epidemiology of this pathogen.     

条斑紫菜6个品系的SRAP分析

为鉴别条斑紫菜不同品系的种质,使用相关序列扩增多态性(sequence-related amplified polymorphism,SRAP)标记对条斑紫菜的5个选育品系和1个野生品系进行遗传分析,结果从35对引物组合中筛选出可扩增出稳定清晰条带的组合11对,共获得131个扩增位点,其中多态性位点125个,多态性比例高达95.42%。6个品系 间的遗传距离为0.364 3～0.867 9,平均为0.593 0。用UPGMA法进行聚类分析,结果将6个品系分为2个群,所反映的亲缘关系与各品系的来源基本一致,说明SRAP 标记技术可以成为条斑紫菜品系间遗传分析的有效工具。在131个多态性位点中,选择扩增出的4个位点构建了6个品系的指纹图谱。另外,通过ME1/EM6引物组合 扩增得到耐高温品系TM-18的特异性条带,经回收测序和重新设计引物,该条带在其丝状体和叶状体DNA中均能稳定地被扩增出来,可用于该品系的种质鉴别 。     

根据mtDNA控制区序列分析野生唇 的种群遗传结构

为了掌握不同地理种群唇(鱼骨)(Hemibarbu labeo)的遗传多样性和遗传结构,探讨其亲缘地理演化过程,研究分析了乌苏里江、长江、黑龙江、鸭绿江和牡丹江5大水系,8个地理群体(n=42)的mtDNA控制区404 bp片段的变异,该片段共有26个变异位点,变异率为6.43%,平均核苷酸多样性为0.011 5.42尾样本共检测到18个单倍型,这8个地理群体是以HT1和HT2单倍型为中心向外辐射演化的.AMOVA分析结果表明,群体内变异为59.29%,同一水系群体间变异为23.50%,不同水系群体间变异为17.20%,以上变异均达到显著水平(P＜0.05),群体间配对FST分析结果表明,同一水系内有些群体间差异也达到了显著水平.聚类分析发现,最大简约法(MP)和最大似然率法(ML)结果基本一致,嘉荫群体(HRJY)和四川群体(YRSC)绝大部分个体被聚在一支,鸭绿江群体(YRLJ)聚在一支,这与单倍型网络的分析结果一致,以上结果表明唇(鱼骨)的遗传多样较为丰富,黑龙江流域的唇(鱼骨)保持了祖先单倍型,其他地理群体为黑龙江流域内群体向不同方向演化的结果,并且部分地理群体已经形成了特有的单倍型.     

基于GBS技术的黄渤海沿岸托氏琩螺遗传特征分析

为了探讨辽宁沿海非经济贝类野生种群的遗传多样性水平和遗传分化特征,本研究选取了托氏琩螺(Umbonium thomasi),利用基于简化基因组测序的基因分型技术(GBS)对辽宁黄渤海沿岸的5个群体进行了全基因组水平的SNP标记开发和遗传特征分析。本研究共获得1315987个SNP标记,发现这5个托氏琩螺群体的观测杂合度和期望杂合度分别为0.0851～0.1161和0.1424～0.1627,观测杂合度均低于期望杂合度,说明托氏琩螺群体存在杂合子缺失,有种群退化风险。C4-ZH群体可能因为外来物种的入侵而发生种群缩小和多样性降低。遗传分化研究发现, 5个托氏琩螺群体处于中等分化水平,其中C5-JZ群体因为区域保护原因与其他4个群体的分化程度相对较高,且通过群体遗传结构分析发现,C5-JZ群体与其他群体的基因交流较少,基于此本研究开发了3个C5-JZ群体的特征性SNP标记。本研究结果将有助于了解辽宁沿海非经济物种的多样性水平,为自然资源的保护和利用提供依据。     

Genetic diversity and structure of Northern Sheatfish (Silurus soldatovi) assessed by newly developed and cross‐species microsatellites

Twenty‐five polymorphic microsatellite markers, including 13 newly isolated markers and 12 existing cross‐species markers, were used to assess the genetic variation of 141 individuals from three cultured stocks of northern sheatfish, Silurus soldatovi, in China. Samples were collected from Heilongjiang (H), Tongerpu (T) and Liaoning (L). Allelic richness, expected and observed heterozygosity among 25 loci were calculated, and the mean A_e, H_o and H_e were estimated in three stocks. In low polymorphic locus HLJcf19 and HLJcf38, most individuals in all three stocks were homozygous and had only one same allele, while only few heterozygous individuals had the other allele in one certain stock. And all the three stocks showed excess of heterozygotes, because of significant negative F_IS values. Genetic distance, UPGMA dendrogram, as well as structure analysis, all indicated that Stock H and L were closer among three stocks. The results revealed that though all stocks had no reduction in recent time, their effective population size were significantly lower to the ideal value.     

Genetic parameters for growth‐related traits and survival with age in the Kuruma shrimp Marsupenaeus japonicus

Genetic parameters were estimated for growth‐related traits and survival of the Kuruma shrimp Marsupenaeus japonicus based on 66 families, including 30 paternal half‐sib families, which were obtained using artificial insemination of two females by each male. The variance components for growth‐related trait and survival were estimated using a single‐trait animal model and a sir‐dam model, respectively, and genetic parameters were estimated using the restricted maximum likelihood method. The coefficients of variation for growth‐related traits such as body length (BL), abdominal length (AL), and body weight (BW) were 9.36%–22.44%, 10.61%–21.92%, and 26.47%–58.33%, respectively, at different ages (45, 75, 105, 135, and 165 days). The corresponding heritability estimates for each growth trait were 0.1545–0.1951, 0.1672–0.1905, and 0.1596–0.1934, respectively, all of which were found to be at moderate levels and increased with age. The heritability of survival on day 165 was low (0.003). The genetic correlations among growth traits were positive and high (0.7316–0.9896) at the different ages, suggesting that selection to improve any single growth trait will cause positive responses in other growth traits examined in M. japonicus. The genetic correlations between growth traits and survival were also positive but low (0.005–0.087), which indicated that only selecting for growth traits may not cause a positive correlated response in the survival of the core population. According to the above results, we suggested that growth and survival traits should be taken as improving targets of breeding in M. japonicus. These results provided reference data for selective breeding and multitrait selection of M. japonicus.     

红唇薄鳅2个野生群体的遗传多样性研究

采用9个微卫星DNA标记对长江支流岷江下游厥溪和长江上游干流朱杨溪红唇薄鳅的2个野生群体遗传多样性及遗传分化情况进行了研究。共检测出73条等位基因,其中厥溪群体中有55条等位基因,朱杨溪群体中有69条等位基因,两群体共有等位基因数为51条。厥溪群体和朱杨溪群体的平均等位基因数分别为6.111和7.667,平均观测杂合度分别为0.665和0.668;平均期望杂合度分别0.684和0.712,平均多态性信息指数分别为0.621和0.656,从各参数结果来看,朱杨溪群体的遗传多样性水平高于厥溪。AMOVA结果显示,大多数遗传变异存在于红唇薄鳅群体内(98.68%),群体间的遗传变异为1.32%,固定系数不显著。基因流为9.42,表明两群体间基因交流频繁。UPGMA聚类显示,厥溪和朱杨溪群体中的个体相互混杂。这些结果表明长江上游厥溪和朱杨溪群体未出现遗传分化。     

黑龙江和长江散鳞镜鲤群体的种质差异分析

本文检测分析黑龙江和长江地区散鳞镜鲤Cyprinus carpio群体的主要生物学特征、生长性能、生化遗传特性。结果表明:长江群体后代鳞被分离较大;全长/体长、体长/体高、体长/体厚、体长/头长、体长/头高、头长/头高、尾柄长/尾柄高等可量性状差异显著。黑龙江群体较长江群体在1龄鱼阶段具有生长优势,2龄鱼生长无明显差异。2个群体1龄和2龄鱼的成活率无明显差异。测量不同酶在不同组织中的活性,结果发现长江散鳞镜鲤与黑龙江散鳞镜鲤在酯酶和乙醇脱氢酶的谱带活性具有一定的差异,淀粉酶、超氧化歧化酶、苹果酸脱氢酶及其他酶的带型和活性差异不明显。