2026· Methods in molecular biology· Vol 3017, pp.
47-57
· 0 citations
Medicine
TL;DR
This protocol presents a step-by-step approach for generating bispyribac sodium-tolerant rice using the CRISPR/Cas-mediated editing of the ALS gene, and offers a robust framework for producing HT rice lines, potentially reducing reliance on conventional herbicide regimes and mitigating the risk of resistance development in weed populations.
Overall, the evidence suggests that CRISPR Cas-9 has the potential to accelerate plant breeding, improve food security, and support the development of climate resilient crops while reducing the time and uncertainty associated with traditional breeding methods.
Victoria Imafidor· World Journal of Advanced En...· 0 citations
This review provides a comprehensive synthesis of a recent advances in CRISPR–Cas technologies and their strategic applications in crop genetics and hybrid breeding, and showcases how these technologies accelerate hybrid breeding by engineering male sterility systems, fixing heterosis, and generating high-throughput mutant libraries for trait discovery.
Syed Riaz Ahmed, Jahangir Khan, I. Ijaz et al.· Frontiers in Plant Science· 0 citations
The results demonstrate the successful deployment of CRISPR/Cas9 for targeted genome engineering in sugarbeet and establish a reliable platform for future gene-editing efforts aimed at enhancing resistance to a wide range of pathogens and diseases affecting the crop.
Z. Khan, Tinley Hathaway, C. Chu et al.· Frontiers in Genome Editing· 0 citations
This critical narrative review evaluates the evidence available across four distinct intervention pathways: editing crops for selective herbicide tolerance; editing crop hosts to resist parasitic weeds; editing weeds for functional genomics or direct fitness manipulation; and developing gene-drive systems for population suppression or restoration of herbicide susceptibility.
M. Karthik, V. Chandrika, D. Subramanyam et al.· Journal of Scientific Resear...· 0 citations
INTRODUCTION
Weeds significantly threaten rice production. While herbicides are the most efficient weed-control method, prolonged use accelerates resistant weed emergence. 4-Hydroxyphenylpyruvate dioxygenase (HPPD)-inhibiting herbicides offer potential for managing resistant weeds in rice, but the availability of resistant rice mutants is limited.
OBJECTIVES
This study aimed to optimize the method of DNA shuffling to conduct directed evolution of the rice HPPD gene, discover novel resistance mutations, and generate herbicide-resistant rice germplasm.
METHODS
We developed an efficient DNA shuffling method utilizing flanking sequences and segmented amplification to construct random mutation libraries of rice HPPD and maize (Zea mays L.) HPPD genes. Mutant libraries were screened via chromogenic reaction in Escherichia coli. CRISPR-mediated adenine base editing was adopted to generate rice mutants, and hydroponic assays were conducted to evaluate herbicide resistance. The crystal structure of rice HPPD was determined by X-ray crystallography, while mutant structures and ligand docking were simulated with AlphaFold3.
RESULTS
Flanking sequences enhanced single-stranded DNA amplification efficiency, and segmented amplification reduced nonsense mutations. A resistant mutant, OsHPPD-mHelix, was obtained. Enzyme kinetics analysis demonstrated that its Ki for mesotrione was increased approximately 2.5-fold compared to that of wild-type OsHPPD, while its enzyme activity was not significantly affected. Using the mutation sites contained in OsHPPD-mHelix as hotspots, genome-edited rice was further generated. It was found that the mesotrione resistance of rice carrying the E423G mutation was approximately 2.2-fold higher than that of wild-type rice. Structural and molecular dynamics analyses suggested the E423G mutation promotes a closed terminal α-helix conformation, with free energy landscapes indicating reduced mesotrione binding stability, potentially explaining the resistance mechanism.
CONCLUSION
This study provides an efficient DNA shuffling technology and novel herbicide-resistant rice germplasm, enriching the toolbox for rice gene-directed evolution and resistant variety cultivation.
Le Chen, Dongshu Guo, Zhou Yang et al.· Journal of Advanced Research· 0 citations
A complete CRISPR-Cas9-mediated knockout of the BEL5 gene, encoding a transcription factor, is reported, known as one of the key regulators driving tuber formation, and a regulatory role of BEL5 in the timing of tuber onset but, unexpectedly, its dispensability for tuber development in modern cultivated potato is proposed.
Andrea Zounková, Daniele Chirivì, A. Přibylová et al.· bioRxiv· 0 citations
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