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| 1 | Advancements in plant regeneration and genetic transformation of grapevine(Vitis spp.)显示文摘Grapevine(Vitis spp.)is one of the most economically important fruit crops worldwide,and there is considerable interest in improving its major agronomic and enological traits in response to ever-changing agricultural environments and consumer demands.Molecular genetic techniques in particular,associated with rapid technological advancements,provide an attractive alternative to conventional breeding approaches for developing new grapevine varieties with enhanced yield performance,quality,stress tolerance and disease resistance.To date,several grapevine varieties have been transformed with genes associated with diverse functions through biolistic bombardment and/or Agrobacterium-mediated transformation,and transgenic grape lines have been obtained using established regeneration systems.Nevertheless,a wide range of factors,including genotype,explant source and culture medium,have been shown to affect the efficiency of plant regeneration.Moreover,the selection and use of acceptor materials,bacterial strain and cell density,selectable markers and selection methods also influence transformation efficiency.This paper provides an overview of recent advances in grapevine regeneration and genetic transformation and in-depth discussion of the major limiting factors,and discusses promising future strategies to develop robust plant regeneration and genetic transformation in grapevine. | ZHANG Xiu-ming WU Yi-fei LI Zhi SONG Chang-bing WANG Xi-ping | 2021 | Journal of Integrative Agriculture2021，20，6： | 1 |
| 2 | Whole-genome sequencing reveals rare off-target mutations in CRISPR/Cas9-edited grapevine显示文摘The CRISPR(clustered regularly interspaced short palindromic repeats)-associated protein 9(Cas9)system is a powerful tool for targeted genome editing,with applications that include plant biotechnology and functional genomics research.However,the specificity of Cas9 targeting is poorly investigated in many plant species,including fruit trees.To assess the off-target mutation rate in grapevine(Vitis vinifera),we performed whole-genome sequencing(WGS)of seven Cas9-edited grapevine plants in which one of two genes was targeted by CRISPR/Cas9 and three wild-type(WT)plants.In total,we identified between 202,008 and 272,397 single nucleotide polymorphisms(SNPs)and between 26,391 and 55,414 insertions/deletions(indels)in the seven Cas9-edited grapevine plants compared with the three WT plants.Subsequently,3272 potential off-target sites were selected for further analysis.Only one off-target indel mutation was identified from the WGS data and validated by Sanger sequencing.In addition,we found 243 newly generated off-target sites caused by genetic variants between the Thompson Seedless cultivar and the grape reference genome(PN40024)but no true off-target mutations.In conclusion,we observed high specificity of CRISPR/Cas9 for genome editing of grapevine. | Xianhang Wang Mingxing Tu Ya Wang Wuchen Yin Yu Zhang Hongsong Wu Yincong Gu Zhi Li Zhumei Xi Xiping Wang | 2021 | Horticulture Research2021，8，1： | 1 |
| 3 | 金银花MLO家族全基因组序列鉴定及表达分析显示文摘通过全基因组测序获得金银花MLO家族候选基因25个。生物信息学分析发现,该家族蛋白氨基酸数量为137~846,理论等电点在5.02~9.50之间,富含碱性氨基酸,有1个蛋白不含跨膜结构域,其他蛋白为3~10个。亚细胞定位预测结果显示,有21个蛋白定位于细胞膜上,1个定位于叶绿体。通过对金银花、小麦、拟南芥、番茄、普通烟草、林烟草和绒毛状烟草等物种的133个MLO蛋白构建系统发育树,显示金银花MLO家族蛋白可分为5个亚组。组织特异性分析发现,金银花MLO家族基因的表达具有明显的组织特异性。其中8个基因在叶中高表达,2个基因在茎中高表达,2个基因在花中高表达。接种白粉菌后,有4个基因显著上调表达,其中MLO14增加最多,比对照提高了2000多倍。本研究初步解析了金银花白粉病发生相关基因MLO的组成及表达情况,为进一步利用MLO作为靶基因,培育金银花抗白粉病新种质奠定了基础。 | 李小丽 魏丽 刘谦 李佳 张芳 张永清 蒲高斌 | 2021 | 药学学报2021，56，10： | 1 |
| 4 | CRISPR/Cas基因组编辑技术在果树育种中的应用显示文摘CRISPR/Cas系统的出现使植物基因组编辑进入了基因精准编辑的新时代。CRISPR/Cas系统已成功应用于多个果树物种。育种研究统计发现,研究者利用CRISPR/Cas基因组编辑技术编辑了大约45个果树基因,主要用于增强抗病性、延长货架期、提早开花、树形矮化体和增加分枝数等方面的研究。目前,果树基因组编辑大多为CRISPR/Cas9系统产生的插入/缺失突变,导入方式主要为农杆菌(Agrobacterium)介导的稳定转化方式,外源基因的瞬时转化体系少有研究,主要是因为大多数果树中转化效率低限制了其应用。本文概述CRISPR/Cas系统及其相关技术,介绍CRISPR/Cas表达载体系统进入植物基因组中的导入方法,并综述CRISPR/Cas系统在果树研究中的应用情况,分析了存在的问题并对其应用前景进行了展望。本文为果树基因功能研究及分子育种研究工作提供参考。 | 倪海枝 王引 王平 程玉芳 颜帮国 陈方永 | 2023 | 农业生物技术学报2023，31，8： | 0 |
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