Kuluev B R, Mikhailova E V, Kuluev A R, Galimova A A, Zaikina E A, Khlestkina E K
Institute of Biochemistry and Genetics, Ufa Research Center, Russian Academy of Sciences, Ufa, 450054 Russia.
Vavilov All-Russia Institute of Plant Genetic Resources, St. Petersburg, 190000 Russia.
Mol Biol (Mosk). 2022 Nov-Dec;56(6):949-968. doi: 10.31857/S0026898422060155.
The tribe Triticeae includes important agricultural crops, such as bread wheat, durum wheat, barley, rye, and triticale. Research in the field of reverse genetics and genetic engineering of Triticeae received a new impetus as the CRISPR/Cas genome editing system came into broad use. The review describes and analyzes the data on recent advances in genomic editing of cultivated plants of the tribe Triticeae and tools used in the field. The tools most commonly used for genome editing in Triticeae include the codon-optimized Cas9 gene under the control of the maize ubiquitin gene promoter and guide RNAs under the control of Pol III promoters U6 and U3 in one or more binary vectors. Phosphinothricin and hygromycin resistance genes are used as selectable genes. Agrobacterium-mediated transformation and biolistics are performed to obtain genome-edited plants, and immature embryos are used as explants. Approaches developed to overcome the problem of low regenerative capacity of Triticeae include in planta transformation of shoot apical meristems, transformation of microspores and pollen grains, and the use of haploinductors. Bread wheat and barley were subject to genomic editing in the majority of studies published to date, and durum wheat and triticale were recently used in CRISPR/Cas knockout studies of target genes. Further progress in the development of genome editing of cultivated plants of the tribe Triticeae should be aimed at expanding the range of species and varieties involved and overcoming the problems of low regenerative capacity. This will allow genetic modification of elite varieties, which will be in demand in agricultural production.
小麦族包括重要的农作物，如面包小麦、硬粒小麦、大麦、黑麦和小黑麦。随着CRISPR/Cas基因组编辑系统的广泛应用，小麦族反向遗传学和基因工程领域的研究获得了新的推动力。本综述描述并分析了小麦族栽培植物基因组编辑的最新进展以及该领域所使用工具的数据。小麦族中最常用于基因组编辑的工具包括在玉米泛素基因启动子控制下的密码子优化Cas9基因，以及在一个或多个二元载体中由Pol III启动子U6和U3控制的引导RNA。草丁膦和潮霉素抗性基因用作选择基因。通过农杆菌介导的转化和基因枪技术来获得基因组编辑植物，并使用未成熟胚作为外植体。为克服小麦族再生能力低的问题而开发的方法包括茎尖分生组织的体内转化、小孢子和花粉粒的转化以及单倍体诱导剂的使用。在迄今为止发表的大多数研究中，面包小麦和大麦都进行了基因组编辑，最近硬粒小麦和小黑麦也被用于靶基因的CRISPR/Cas敲除研究。小麦族栽培植物基因组编辑的进一步发展应旨在扩大所涉及的物种和品种范围，并克服再生能力低的问题。这将使优良品种能够进行基因改良，而优良品种在农业生产中是有需求的。
