利用CRISPR/CAS9技术定点编辑水稻花粉特异表达基因OsIPA

摘要

水稻雄配子发育是水稻得以繁殖的关键发育过程,同时水稻的雄性不育在水稻杂种优势利用中也发挥着重要作用.花药/花粉特异表达基因可能在雄配子发育中发挥关键作用.本研究利用CRISPR/CAS9技术对水稻品种明恢86中的花药特异表达基因OsIPA定点编辑,获得OsIPA突变体,以期研究OsIPA基因在水稻花粉发育过程中的功能.试验最终成功的对OsIPA基因第1外显子上的2个不同的靶位点进行了定点突变,经检测发现有24株T0代在相应的靶位点发生了突变,共有8种不同的类型,包括碱基缺失、替换以及单碱基的插入等不同类型.%Male gamete development is an essential step in rice reproduction process,and male sterility is crucial for the utilization of rice heterosis.Since the anther or pollen specific genes in rice might play a pivotal role in the male gamete development,the functions of OsIPA were of great interest to the scientists.This study applied CRISPR/CAS9 to generate an OsIPA mutant for an investigation.Two sites in the 1st exon of OsIPA were selected as the targets.After sequencing,a total of 8 different types of mutant,including deletion,substitution and single nucleotide insertion,were found in 24T0 transgenic plants.