Park Jae-Ryoung, Kim Eun-Gyeong, Jang Yoon-Hee, Jan Rahmatullah, Farooq Muhammad, Ubaidillah Mohammad, Kim Kyung-Min
Division of Plant Biosciences, School of Applied Biosciences, College of Agriculture and Life Science, Kyungpook National University, Daegu, South Korea.
Crop Breeding Division, National Institute of Crop Science, Rural Development Administration, Wanju, South Korea.
Front Plant Sci. 2022 Feb 24;13:850441. doi: 10.3389/fpls.2022.850441. eCollection 2022.
Recent unpredictable climate change is the main reason for the decline in rice yield. In particular, drought stress is a major constraint in reducing yield and quality for rice at rainfed agriculture areas, such as Asia and South America. CRISPR/Cas9 provides an effective solution for gene function study and molecular breeding due to specific editing of targeted genome sequences. In addition, CRISPR/Cas9 application can significantly reduce the time required to develop new cultivars with improved traits compared to conventional complex and time-consuming breeding. Here, drought-induced gene () was edited by CRISPR/Cas9. To investigate the possible role of in drought stress, genome-editing plants were subjected to drought stress until the soil moisture content reached 20%, and the reactive oxygen species (ROS) scavenging efficiency of genome-editing plants were decreased. When the genome-editing plants were subjected to drought stress, survival rate, shoot length, root length, content of chlorophyll number of tiller, and 1,000-grain weight decreased, and more HO and O were detected in leaves. In addition, expression levels of several critical stress-related transcription factors were decreased in the genome-editing plant. These results suggest that function as a positive regulator during drought stress response in rice. We analyzed the expression of and in T and T plants as well as T seeds. As the course of generation advancement progressed, expression remained stable or weakened but the expression was continuously removed from the T plant. The coefficient of variation (CV) in both T plants and T seeds was lower than 5%. Overall, our results suggest that CRISPR/Cas9 could be a novel and important tool for efficiently generating specific and inheritable targeted genome editing in rice, with short breeding cycles.
近期不可预测的气候变化是水稻产量下降的主要原因。特别是，干旱胁迫是亚洲和南美洲等雨养农业地区水稻产量和品质降低的主要限制因素。由于对靶向基因组序列进行特异性编辑，CRISPR/Cas9为基因功能研究和分子育种提供了一种有效的解决方案。此外，与传统复杂且耗时的育种相比，CRISPR/Cas9的应用可显著缩短培育具有改良性状新品种所需的时间。在此，利用CRISPR/Cas9对干旱诱导基因（）进行编辑。为了研究该基因在干旱胁迫中的可能作用，对基因组编辑植株施加干旱胁迫直至土壤含水量达到20%，结果发现基因组编辑植株的活性氧（ROS）清除效率降低。当对基因组编辑植株施加干旱胁迫时，其存活率、地上部长度、根长度、叶绿素含量、分蘖数和千粒重均下降，并且在叶片中检测到更多的HO和O。此外，在该基因的基因组编辑植株中，几个关键的胁迫相关转录因子的表达水平降低。这些结果表明，该基因在水稻干旱胁迫响应过程中作为正向调节因子发挥作用。我们分析了该基因在T和T植株以及T种子中的表达情况。随着世代推进，该基因的表达保持稳定或减弱，但该基因的表达在T植株中持续消除。T植株和T种子的变异系数（CV）均低于5%。总体而言，我们的结果表明，CRISPR/Cas9可能是一种新颖且重要的工具，可用于在水稻中高效产生特异性且可遗传的靶向基因组编辑，育种周期短。
