Applications of CRISPR/Cas9 as New Strategies for Short Breeding to Drought Gene in Rice

Abstract

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 Oryza sativa Senescence-associated protein (OsSAP) was edited by CRISPR/Cas9. To investigate the possible role of OsSAP 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 H2O2 and O2− were detected in leaves. In addition, expression levels of several critical stress-related transcription factors were decreased in the OsSAP genome-editing plant. These results suggest that OsSAP function as a positive regulator during drought stress response in rice. We analyzed the expression of OsSAP and Cas9 in T0 and T1 plants as well as T2 seeds. As the course of generation advancement progressed, Cas9 expression remained stable or weakened but the OsSAP expression was continuously removed from the T0 plant. The coefficient of variation (CV) in both T1 plants and T2 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.

Salt and Heat Stress Trigger Morpho-Physiological Changes, Antioxidant Enzyme and Secondary Metabolites Gene Expression in Rice (Oryza sativa L.)

“…Several antioxidant genes analyzed in this study could trigger the formation of antioxidant enzymes, namely superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione peroxidase (GPOD). Due to stress conditions, these enzymes play a role in the ROS scavenging process (Kibria et al 2017;Park et al 2022).…”

Unveiling the protective role of anthocyanin in rice: insights into drought-induced oxidative stress and metabolic regulation

“…After successful sprouting the seedling were transferred to pots and kept in dark condition for three days. The experiment was conducted in green house at 28/26°C and 16/8 light/dark condition ( Park et al., 2022 ). The 5 µM anthocyanin was applied to one month old plant, one week before to drought stress exposure.…”

Advances in Environmental Biotechnology with CRISPR/Cas9: Bibliometric Review and Cutting-Edge Applications

“…The scope of CRISPR/Cas9's application extends to the development of improved fruit, vegetable, and ornamental crops, as illustrated by Erpen-Dalla et al (2019) [118], and the conferment of enhanced resistance to rice root-knot nematode, a significant threat in rice agriculture [119]. Innovations in floricultural crops through genetic improvement [120], drought gene breeding in rice [121], and the enhancement of horticultural crops [122] further showcase the technology's versatility.…”