【福利】最新摘要发布-2017植物基因组与基因编辑学术研讨会

基因编辑是一种用于精确操纵细胞和生物体内基因和基因组的技术。近年来正迅速影响着植物基因组学研究领域，逐渐受到学术界及全社会的关注。生物谷在将于11月10日在上海好望角大酒店举办《2017植物基因组与基因编辑学术研讨会》。为大家提供一个交流技术和前沿信息的平台，推动植物基因编辑领域的发展。
 
以下是小编为大家带来的福利哦，部分与会嘉宾的摘要分享！
 
 
【陈其军】中国农业大学生物学院

演讲题目：高特异性和高通量植物CRISPR/Cas9基因组和碱基编辑技术应用研究

1.从另一个视角（文献计量学）分享CRISPR革命浪潮的气势；
2.报告我们建立的基因组和碱基编辑工具箱；
3.报告我们在研究过程中遇到的一序列意外的发现，包括在拟南芥中的高频率脱靶现象以及在下一代脱靶加重现象；
4.报告高特异性Cas9在植物中的效率及特异性；
5.介绍使用两套互补干扰CRISPR系统实现拟南芥多基因敲除的途径；
6.介绍实现玉米高通量基因组和碱基编辑的途径。
 
 
 
【金双侠】华中农业大学植物科学技术学院

演讲题目：多倍体棉花的基因组编辑研究进展

As one of gene editing technologies, the CRISPR/Cas9 system has exerted its broad application from prokaryotes to eukaryotes. Its robustness, costlessness and high efficiency give it obvious advantages and potential compared to both ZFN and TALENs gene editing technologies. Up to now, it had conspicuous effects in many plants, including major crops like rice, wheat and zea maize. As an important economic crop, the widely cultivated upland cotton is an allotetraploid with a complex genome structure and most genes have at least two copies that originated from A and D sub-genome, respectively. This feature always results in no obvious phenotype in gene functional analysis by RNA interference strategy, because of gene redundancy. There are no sufficient mutants in cotton for advanced gene function research. In this study, we utilized a CRISPR/Cas9 system that can generate more than one sgRNAs in one vector for genome editing at multiple sites. An exogenously transformed gene dsRFP and an endogenous gene GhCLA1 were chose as CRISPR/Cas9 knock out targets. For each gene, at least a pair of sgRNAs, driven by a cotton native type Ⅲ RNA polymerase promoter-pGhU6.9, were designed in the coding (or exons) regions. Through Agrobacterium-mediated transformation, the regenerated T0 plants exhibited obvious phenotype for both target genes. For RFP, the regenerated plants had vanished red fluorescence. GhCLA1 targeting plants gained obvious albefaction from embryo to young seedlings stages. Gene mutations were verified with Sanger sequencing and T7E1 enzyme. Large deletions were observed between the paired target sites. Moreover, these albefaction plants only arose in bioallelic mutation of all copies of the target gene. All these results demonstrated that the sgRNA guided Cas9 had a great potential in cotton for genome editing.  

 
 
【徐明良】中国农业大学

演讲题目：玉米抗病QTL克隆及标记辅助抗性改良

玉米病害是玉米生产的主要限制因子，每年造成的产量损失约占总产的10%。近年来，由于极端气候频发，秸秆还田、机械化收获等因素的影响，我国玉米病害呈越演越烈之势。有别于禾谷类其他作物，如水稻、玉米，大多数重要玉米病害都是由坏死营养型病原菌引起，相应的玉米抗病性呈数量性状遗传，由多个抗病位点控制。虽然数量性状抗病位点只能使抗性适度提高，但这类抗性稳定，广谱且可持续，在玉米育种中有重要的利用价值。如果将多个数量抗性位点聚合可以达到对某个病害高度的抗性。为了从遗传上改良玉米的抗病性，我们与玉米育种家合作，收集、筛选优良抗源，组配定位群体，开展图位克隆。迄今为止，我们已克隆了9个重要玉米抗病QTL基因，分别抗丝黑穗病、茎腐病、穗粒腐病、甘蔗花叶病毒病、粗缩病、灰斑病等。根据抗病基因序列信息，我们在玉米种质资源中挖掘优良抗病等位基因，发展分子标记，利用分子标记辅助选择将优良抗病等位基因导入到玉米自交系中，培育抗性改良的优良自交系。目前，我们发掘的优良抗病基因已广泛用于我国玉米抗病育种。
 
 
【刘克德】华中农业大学

演讲题目：
CRISPR/Cas9-mediated genome editing efficiently creates specific mutations at multiple loci using one sgRNA in Brassica napus

Hong Yang1, Jia-Jing Wu1, Ting Tang1, Ke-De Liu1*, Cheng Dai1*
1. National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070, China
Abstract
CRISPR/Cas9 is a valuable tool for both basic and applied research that has been widely applied to different plant species.  Nonetheless, a systematical assessment of the efficiency of this method is not available for the allotetraploid Brassica napus—an important oilseed crop. In this study, we examined the mutation efficiency of the CRISPR/Cas9 method for 12 genes and also determined the pattern, specificity and heritability of these gene modifications in B. napus. The average mutation frequency for a single-gene targeted sgRNA in the T0 generation is 65.3%. For paralogous genes located in conserved regions that were targeted by sgRNAs, we observed mutation frequencies that ranged from 27.6% to 96.6%. Homozygotes were readily found in T0 plants. Meantime, we developed a simple visual screen for mutant material in plant species, allowing us to quickly obtain mutated plants or tissue. We observed mutation frequencies that ranged from 7.4% to 100% in Arabidopsis and B. napus, respectively. Cas9-free mutants were sufficiently isolated from T2 generation in Arabidopsis and B. napus by visual screening. Homozygotes, bi-alleles, and heterozygotes were stably inherited as classic Mendelian alleles in the next generation (T3) without any new mutations or reversions. Moreover, no mutation was found in the putative off-target sites among the examined plants or tissue. The method allows us for effectively isolating positive transgenic plants and Cas9-free heritable CRISPR mutants in many plant species, which open many doors for biotechnological applications in oilseed crops.

Keywords: CRISPR/Cas9, gene editing, B. Napus.

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