CRISPR-Cas system, a natural acquired immune system in prokaryotes that defends against exogenous DNA invasion because of its simple structure and easy operation, has been widely used in many research fields such as synthetic biology, crop genetics and breeding, precision medicine, and so on. The miniature CRISPR-Cas12 system has been an emerging genome editing tool in recent years. Compared to the commonly used CRISPR-Cas9 and CRISPR-Cas12a, the miniature CRISPR-Cas12 system has unique advantages, such as rich PAM sites, higher specificity, smaller volume, and cytotoxicity. However, the application of miniature Cas12 proteins and the methods to improve its editing efficiency have not been systematically summarized. In this review, we introduce the classification of CRISPR-Cas system and summarize the structural characteristics of type V CRISPR-Cas system and the cleavage mechanism of five miniature Cas12 proteins. The application of a miniature CRISPR-Cas12 system in the gene editing of animals, plants, and microorganisms is summarized, and the strategies to improve the editing efficiency of the miniature CRISPR-Cas12 system are discussed, aiming to provide reference for further understanding the functional mechanism and engineering modification of the miniature CRISPR-Cas12 system.

Cadmium (Cd) is a heavy metal that poses harm to both plants and humans. OsHMA3, a member of the heavy metal ATPase (HMA) family, plays a crucial role in sequestering Cd into the vacuoles of roots, thereby limiting its accumulation in rice grains. However, the response of rice plants to Cd under complete loss-of-function of OsHMA3 remains unclear. In this study, we successfully generated OsHMA3 null mutants in an indica variety 93 − 11 using CRISPR/Cas9 technology. A Cd resistance experiment revealed that the Oshma3 mutants exhibited increased sensitivity to Cd compared to the wild-type at a tested concentration of 10 µM CdCl₂. Furthermore, the seedlings of Oshma3 mutant lines displayed inhibited plant growth in the presence of 1 µM Cd, specifically suppressing aboveground growth. As expected, knockout lines of OsHMA3 showed lower Cd accumulation in roots but higher concentrations in shoots compared to wild-type plants, highlighting the role of OsHMA3 in root-to-shoot Cd translocation. We further performed RNA sequencing analysis on wild-type and Oshma3 plants under control and Cd treatment conditions and found that differentially expressed genes were mainly enriched in metal ion binding, integral component of the membranes, and biosynthesis pathways for secondary metabolites triggered by exposure to Cd. When grown in a paddy field, the Oshma3 mutants exhibited shorter plant height, lower seed setting rate, and higher Cd accumulation in grains compared to wild-type plants. Our results indicate that knockout of OsHMA3 in the 93 − 11 variety increases sensitivity to Cd and inhibits plant growth.

As a staple food crop, Oryza sativa L. is not only the basis of global food and nutrition security but also an important cornerstone of national economic development and social stability. However, the growth of rice is often accompanied by the threat of rice blast, which can lead to the death of seedlings or plants before heading. In the later stages of growth, a severe leaf blast infection will reduce the leaf area at the filling stage, thereby reducing the grain yield. The study of rice blast resistance genes and susceptibility genes is a key strategy for controlling the occurrence of rice blast and ensuring sustainable rice production. This paper reviews the impact of rice blast on the global economy and food security in recent years, describes the immune mechanism of rice blast resistance, and introduces the latest progress in related research. At the same time, the main genes of rice blast resistance and the resistance-related genes, as well as the susceptibility genes identified or cloned in recent years, are summarized. This paper also discusses the application of conventional breeding, molecular-marker-assisted breeding, gene editing, and other technologies in rice blast resistance breeding. The problem of accurately finding avirulence genes for R genes in current disease-resistant breeding is discussed and explored, aiming to improve rice blast resistance, agronomic traits, and yield in a sustainable way.

Aluminum (Al) stress is a dominant obstacle for plant growth in acidic soil, which accounts for approximately 40–50% of the world’s potential arable land. The identification and characterization of Al stress response (Al-SR) genes in Arabidopsis, rice, and other plants have deepened our understanding of Al’s molecular mechanisms. However, as a crop sensitive to acidic soil, only eight Al-SR genes have been identified and functionally characterized in maize. In this review, we summarize the Al-SR genes in plants, including their classifications, subcellular localizations, expression organs, functions, and primarily molecular regulatory networks. Moreover, we predict 166 putative Al-SR genes in maize based on orthologue analyses, facilitating a comprehensive understanding of the impact of Al stress on maize growth and development. Finally, we highlight the potential applications of alleviating Al toxicity in crop production. This review deepens our understanding of the Al response in plants and provides a blueprint for alleviating Al toxicity in crop production.

Mitogen-activated protein kinase (MAPK/MPK) cascades are pivotal and highly conserved signaling modules widely distributed in eukaryotes; they play essential roles in plant growth and development, as well as biotic and abiotic stress responses. With the development of sequencing technology, the complete genome assembly of rice without gaps, T2T (Telomere-to-Telomere)—NIP (version AGIS-1.0), has recently been released. In this study, we used bioinformatic approaches to identify and analyze the rice MPK kinases (MKKs) based on the complete genome. A total of seven OsMKKs were identified, and their physical and chemical properties, chromosome localization, gene structure, subcellular localization, phylogeny, family evolution, and cis-acting elements were evaluated. OsMKKs can be divided into four subgroups based on phylogenetic relationships, and the family members located in the same evolutionary branch have relatively similar gene structures and conserved domains. Quantitative real-time PCR (qRT-PCR) revealed that all OsMKKs were highly expressed in rice seedling leaves. The expression levels of all OsMKKs were more or less altered under exogenous hormone and abiotic stress treatments, with OsMKK1, OsMKK6, and OsMKK3 being induced under almost all treatments, while the expression of OsMKK4 and OsMKK10-2 was repressed under salt and drought treatments and IAA treatment, respectively. In this study, we also summarized the recent progress in rice MPK cascades, highlighted their diverse functions, and outlined the potential MPK signaling network, facilitating further studies on OsMKK genes and rice MPK cascades.