芸薹属及其他异源多倍体植物的基因表达特征研究进展

远缘杂交及异源多倍化导致许多重要作物的起源与进化，而芸薹属栽培异源四倍种是研究作物异源多倍化的模式系统之一。异源多倍体是如何调控及协调来自不同二倍体祖先的不同基因组的遗传行为及基因表达，是过去二十年间的研究热点和重要的生物学问题。利用不断发展的分子生物学技术，一方面揭示出芸薹属及其他多倍体物种基因组表现出动态性质，即在形成初期及长期进化过程中持续发生遗传及表观遗传的变化；另一方面发现异源多倍化过程中伴随着大量的基因表达模式改变，包括非加性表达、超亲表达、表达水平显性、部分同源偏向表达、基因剂量平衡效应等现象。上述基因组结构、表观遗传改变以及基因表达模式的调控，使新产生的多倍体得以成功进化为新物种。     

Genetic Engineering Approaches to Pig Production*

Modern biotechnology promises a number of new applications in animal breeding and production. Although conventional pig breeding has achieved a high level of efficiency and productivity numerous problems have been encountered with animal health and the loss of meat quality. Selection based on phenotypic performance data of individual animals does not take into account the importance of specific genes and their relevance within a complex regulatory system. In most cases it is therefore difficult to trace back the genetic origins of clinically important disorders. The application of genetic engineering techniques in pig production will facilitate diagnosis, improvement of productivity, and animal health by allowing direct genetic manipulation. Attention must be focussed on the physical and genetic analysis of the procine genome. The isolation and characterisation of genes, DNA-markers, polymorphic DNA-fragments, and their chromosomal assignment will be important prerequisites and tools for the elucidation of genetic disorders. Especially the detection of heterozygous carriers of recessive disorders and their elimination from the breeding stock will increase selection accuracy and decrease the generation intervals. But also the rapid and simple detection of infectious diseases, which is sometimes difficult if not impossible at present, will improve animal health and welfare. Although the production of transgenic animals either by DNA-microinjection into zygotes or the use of embryonal stem cells manipulated in vitro is less straightforward than DNA-based diagnosis it will play an important role in the direct manipulation of the porcine genome and genes. Breeding programmes including the use of transgenic livestock have already been developed. There is no doubt that genetic engineering has reached a degree of practical feasibility, allowing it to play an important role in pig breeding in particular and animal production in general.     

苹果基因组分子生物学研究进展

借助分子标记手段苹果基因组研究取得了巨大成就,涉及苹果的抗性、生长发育、果实品质等各方面基因,综合有关文献,将其研究的主要成果,包括已发现的主要苹果遗传标记、已构建的苹果遗传连锁图谱及苹果的转基因研究进行综述,并对目前苹果基因组研究存在的问题和对策进行了分析,以期为苹果遗传育种提供参考。     

Identification of a class of CFEM proteins containing a new conserved motif in Fusarium oxysporum

Fusarium oxysporum (Fo) is an important soil-borne fungus that cause huge economic losses worldwide. Fungal-specific Common in Fungal Extracellular Membrane (CFEM) proteins are known to be involved in some important physiological processes associated with pathogenicity. To date, few Fo CFEM proteins have been characterized. The recent publication of several genomes of Fo has allowed us to conduct a genome-wide comparison analysis of CFEM proteins in Fo. In this study, we identified CFEM proteins for 12 different Fo formae speciales(f. sp) and obtained an average of 16 CFEM proteins for each Fo. The Fo CFEM proteins were classified into three groups (groups 1–3) according to structural features. Importantly, we identified a new conserved motif containing about 50 amino acids (DR motif) in group 1 members. CFEM proteins containing DR motif (CFEM_DR proteins) are remarkably conserved among Fo, and their number is greater in Fo compared with other fungi. We found the expansion of CFEM_DR proteins in Fo can be attributed to the segmental duplications in the genomes. Expression analysis revealed that CFEM_DR genes had a higher expression level in mycelium than conidia, and their expressions increased dramatically in the host roots at 3 days post inoculation, indicating that Fo CFEM_DR genes have roles while colonizing and infecting their hosts.     

Improvements of TKC Technology Accelerate Isolation of Transgene-Free CRISPR/Cas9-Edited Rice Plants

Elimination of the CRISPR/Cas9 constructs in edited plants is a prerequisite for assessing genetic stability, conducting phenotypic characterization, and applying for commercialization of the plants. However, removal of the CRISPR/Cas9 transgenes by genetic segregation and by backcross is laborious and time consuming. We previously reported the development of the transgene killer CRISPR (TKC) technology that uses a pair of suicide genes to trigger self-elimination of the transgenes without compromising gene editing efficiency. The TKC technology enables isolation of transgene-free CRISPR-edited plants within a single generation, greatly accelerating crop improvements. Here, we presented two new TKC vectors that show great efficiency in both editing the target gene and in undergoing self-elimination of the transgenes. The new vectors replaced the CaMV35S promoter used in our previous TKC vector with two rice promoters to drive one of the suicide genes, providing advantages over our previous TKC vector under certain conditions. The vectors reported here offered more options and flexibility to conduct gene editing experiments in rice.     

利用CRISPR/Cas9系统编辑日本结缕草ZjSGR基因技术研究

日本结缕草(Zoysia japonica)是一种常见的具有众多优良性状的暖季型草种,秋冬季节在我国北方地区会较早出现叶片脱绿现象,在一定程度上限制了日本结缕草在北方的大面积使用。ZjSGR是从结缕草中分离出来的一种衰老诱导基因,负调控叶片滞绿性状。本研究利用CRISPR/Cas9系统,通过基因枪介导的遗传转化方法实现对日本结缕草ZjSGR基因的编辑,经测序验证获得1株单碱基缺失突变植株。突变植株叶片深绿,在正常生长温度(28~30℃)条件下,和对照相比,叶绿素含量更高(P<0.05)。本研究为进一步阐述ZjSGR基因在日本结缕草滞绿性状调控中的作用机理提供了基础研究材料,也为培育日本结缕草滞绿品种育种工作奠定了基础。     

Genome-edited crops: how to move them from laboratory to market

Recent breakthroughs in CRISPR technology allow specific genome manipulation of almost all crops and have initiated a revolution in precision crop breeding. Rationally-based regulation and widespread public acceptance are needed to propel genome-edited crops from laboratory to market and to translate this innovative technology into agricultural productivity.     

Whole-genome sequence of a novel Chinese cyprinid herpesvirus 3 isolate reveals the existence of a distinct European genotype in East Asia

Cyprinid herpesvirus 3 (CyHV3), also known as koi herpesvirus (KHV), can be subdivided primarily into European and Asian genotypes, which are represented by CyHV3-U or CyHV3-I and CyHV3-J, respectively. In this study, the whole genome sequence of a novel Chinese CyHV3 isolate (GZ11) was determined and annotated. CyHV3-GZ11 genome was found to contain 295,119 nucleotides with 52.9% G/C content, which is highly similar to those of published CyHV3-U, CyHV3-I, and CyHV3-J strains. With reference to CyHV3-U, CyHV3-I, and CyHV3-J, CyHV3-GZ11 was also classified into 164 open reading frames (ORF), which include eight repeated ORFs. On the basis of the 12 alloherpeviruses core genes, results from phylogenetic analysis showed that CyHV3-GZ11 had closer evolutionary relationships with CyHV3-U and CyHV3-I than with CyHV3/KHV-J, which were also supported by genome wide-based single nucleotide substitution analysis and the use of a series of developed molecular markers. This study was the first to reveal the presence of a distinct European CyHV3 genotype in East and Southeast Asia at a whole genome level, which will evoke new insights on exploring the origin, evolution, and epidemiology of the virus.     

茶树咖啡碱合成酶CRISPR/Cas9基因组编辑载体的构建

CRISPR/Cas9技术是一门新兴的基因组定点编辑技术,具有操作简单、高效的优点,可轻松实现对目标基因的敲除、替换和定点突变等操作。该技术刚诞生,就受到了全球生命科学领域研究者的关注,不到3年的时间就已经成功应用于多种动、植物当中。然而CRISPR/Cas9技术在茶树中的应用面临载体构建问题,本文以茶树咖啡碱合成酶为例,联合采用常规PCR、Overlapping PCR和Golden Gate Cloning技术,构建了包含茶树咖啡碱合成酶双靶点的CRISPR/Cas9基因编辑载体,为CRISPR/Cas9介导的基因组编辑技术在茶树中的应用奠定了坚实基础。     

重组减蛋综合征病毒KnobS干酪乳杆菌的构建与表达

以干酪乳杆菌为传递载体,以开发食品级安全的减蛋综合征病毒疫苗为目标,构建了一个温度敏感型自杀型质粒系统p ORZP-UKD。将该质粒电转化到干酪乳杆菌L.casei中,经过2次升温处理和5-氟尿嘧啶抗性筛选,挑选阳性克隆,采用PCR和SDS-PAGE方法进行鉴定。KnobS基因整合到干酪乳酸杆菌基因组内,并实现融合蛋白KnobS的分泌表达。表明得到了一株具有无任何选择性标记、携带有KnobS表达盒元件的基因组整合的重组干酪乳酸杆菌KnobSΔupp L.casei,且整合的外源基因能够随着细菌的复制而进行表达,为减蛋综合征病毒免疫保护性抗原KnobS疫苗的进一步研制提供了试验数据。