CRISPR/Cas-based genome editing for enhancing grain quality traits in rice (Oryza sativa L.)

Genome editing-CRISPR/Cas9 for the crop improvement of Industrial Cannabis sativa (Hemp)

Industrial Cannabis sativa (Hemp) is a multipurpose revolutionary crop widely cultivated for its seeds, which are rich in oil, CBD content, proteins, carbohydrates, fibers, as well as vitamins and minerals. Genome editing, CRISPR/Cas9 acts as highly precise "genetic scissors" that accelerate traditional plant and animal breeding by modifying specific DNA sequences without introducing foreign (transgenic) DNA. The world's first genome-edited rice varieties are DRR Dhan 100 (Kamala) and Pusa DST Rice 1 developed by the Indian Council of Agricultural Research (ICAR),New Delhi, India. Genome editing, CRISPR/Cas9 revolutionizes Cannabis sativa (hemp) breeding by enabling precise, targeted modifications in a crop traditionally hampered by dioecy and high heterozygosity. CRISPR/Cas9 technology allows for sequence specific editing of the target genome, thereby allowing for precise control over gene modifications and associated traits, in a low cost and straightforward manner. Gene editing has been successfully used to generate climate-resilient crops for various climatic conditions. However, several limitations are descending its overall potential remains limited. The biological functions of most candidate genes are poorly characterized. For practical crop improvement in Cannabis sativa, functional validation serves as a bridge between genomic discovery and breeding applications. The Cannabis sativa genome has long been challenging to genetically manipulate due to recalcitrance in tissue culture and transformation, limiting functional validation of genes and targeted trait improvement. The major drawback of gene-editing technologies is off-target, which can cause unwanted editing of other genes, which hinders their wide applicability for crop trait improvement. Despite these advances, transformation and regeneration remain the major bottlenecks limiting widespread genome editing in Cannabis. Another disadvantage is the lack of efficient tissue culture methods for regeneration, transformation and regeneration of gene edited hemp crops. There are studies highlighting successful Cannabis sativa organogenesis but the commercial scale production is still a problem. However, in vitro regeneration of different varieties of hemp is very slow and found recalcitrant. This is another major disadvantage for the application of genome editing, CRISPR/Cas9 in the crop improvement of Industrial Cannabis sativa, hemp. Further limitations are caused by the regulatory uncertainty surrounding genetically edited cannabis.

Transgenic sorghum with improved grain protein digestibility: copy number inheritance of the gKAF1 RNAi genetic construct, and its effects on agronomically important traits

Genome editing in tetraploid potato: CRISPR technologies, multi-omics target discovery, and applications for trait-oriented breeding

This review summarizes examples of reduced tuber browning, modified starch characteristics, and editing of susceptibility loci for late blight and viral resistance, as well as technical challenges specific to potato, such as allele identification in tetraploids, editing efficiency, and bystander edits.

Genome-wide analysis and expression profiling of CPK gene families in Sorghum bicolor (L.) Moench reveal their hormone-mediated responses during seed germination

This study identifies key candidate SbCPK genes associated with seed germination and hormonal signal transduction, providing a fundamental theoretical basis for further elucidating the calcium signaling-mediated molecular regulatory mechanisms of PHS in sorghum.

Multifaceted Effects of Starch Branching Enzyme and Soluble Starch Synthase Gene Editing in Rice with Different Wx Genotypes

Rice endosperm, the major edible portion of the grain, plays an important role in regulating blood glucose and preventing intestinal diseases by increasing its resistant starch (RS) content. Previous studies have shown that suppressing amylopectin biosynthesis via genome editing can increase RS content. However, the influence of different Waxy (Wx) allelic backgrounds on RS accumulation in edited lines has not been systematically evaluated. In this study, we used glutinous rice Yunan Heixiangnuo (HXN) with a nonfunctional wx allele and indica rice Yixiang 1B (YX1B) with a weak Wxb allele as backgrounds. We simultaneously knocked out SSSIIIa, SBEI, SBEIIa, and SBEIIb using CRISPR/Cas9, and systematically analyzed changes in RS content, rice quality, and yield traits. The results showed that, in the HXN background, multigene knockout did not significantly alter amylose or RS content but largely maintained favorable eating quality. In contrast, in the YX1B background, quadruple-gene knockout lines exhibited an increase in amylose content from 17.7% to 53.7% and an increase in RS content to 2.48%, representing a 4.35-fold increase over the wild type, while gel consistency and seed-setting rate were significantly reduced. Scanning electron microscopy revealed that multigene knockout markedly remodeled starch granule structure, shifting from dense polygonal granules to loosely packed spherical particles and resulting in a floury endosperm. Collectively, enhancement of RS content through multigene editing was influenced by Wx gene function. Although the wx allele failed to increase RS content, it still participated in the regulation of grain quality and yield traits by modulating starch structure. This study provides a reference for breeding high-RS rice cultivars while balancing yield performance and eating quality.

CRISPR/Cas9-based gene-editing platform development by targeting genes encoding magnesium chelatase and phytoene desaturase in sugarbeet (Beta vulgaris L.)

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.