Breeding rice for yield improvement through CRISPR/Cas9 genome editing method: current technologies and examples.

The impending climate change is threatening the rice productivity of the Asian subcontinent as instances of crop failures due to adverse abiotic and biotic stress factors are becoming common occurrences. CRISPR-Cas9 mediated genome editing offers a potential solution for improving rice yield as well as its stress adaptation. This technology allows modification of plant's genetic elements and is not dependent on foreign DNA/gene insertion for incorporating a particular trait. In this review, we have discussed various CRISPR-Cas9 mediated genome editing tools for gene knockout, gene knock-in, simultaneously disrupting multiple genes by multiplexing, base editing and prime editing the genes. The review here also presents how these genome editing technologies have been employed to improve rice productivity by directly targeting the yield related genes or by indirectly manipulating various abiotic and biotic stress responsive genes. Lately, many countries treat genome-edited crops as non-GMOs because of the absence of foreign DNA in the final product. Thus, genome edited rice plants with improved yield attributes and stress resilience are expected to be accepted by the public and solve food crisis of a major portion of the globe. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-024-01423-y.

A simplified and improved protocol of rice transformation to cater wide range of rice cultivars.

The latest CRISPR-Cas9-mediated genome editing technology is expected to bring about revolution in rice yield and quality improvement, and thus validation of rice transformation protocols using CRISPR-Cas9-gRNA constructs is the need of the hour. Moreover, regeneration of more number of transgenic rice plants is prerequisite for developing genome-edited rice lines, as recalcitrant rice varieties were shown to have lower editing efficiencies which necessities screening of large number of transgenic plants to find the suitable edits. In the present study, we have simplified the Agrobacterium-mediated rice transformation protocol for both Indica and Japonica rice cultivars using CRISPR/Cas9 empty vector construct, and the protocols have been suitably optimized for getting large numbers of the regenerated plantlets within the shortest possible time. The Japonica transgenic lines were obtained within 65 days and for the Indica cultivars, it took about 76-78 days. We also obtained about 90% regeneration efficiency for both Japonica and Indica cultivars. The transformation efficiency was about 97% in the case of Japonica and 69-83% in the case of Indica rice cultivars. Furthermore, we screened the OsWRKY24 gene editing efficiency by transforming rice cultivars with CRISPR/Cas9 construct harbouring sgRNA against OsWRKY24 gene and found about 90% editing efficiency in Japonica rice cultivars, while 30% of the transformed Indica cultivars were found to be edited. This implicated the presence of a robust repair mechanism in the Indica rice cultivars.

Hepatoprotective effects of Psidium guajava on mitochondrial enzymes and inflammatory markers in carbon tetrachloride-induced hepatotoxicity.

OBJECTIVE: The present investigation was aimed to evaluate the hepatoprotective potential of ethanolic extract of Psidium guajava (P. guajva) and its isolated quercetin fraction on carbon tetrachloride (CCl4)-induced hepatotoxicity. MATERIALS AND METHODS: The rats were divided into 6 groups and each group contained 6 rats. CCl4 (1.5 ml/kg b.w.) was used to induce the hepatotoxicity. Ethanolic extract of P. guajava (300 mg/kg b.w.), isolated quercetin fraction (20 mg/kg b.w.) were used as a treatment and silymarin (25 mg/kg b.w.) was used as a standard drug. After the study period, the liver tissues were collected and evaluate the levels of liver functional markers, mitochondrial enzymes, histopathological analysis and the expressions of inflammatory markers. RESULTS: The levels of liver functional markers were increased and protein, albumin and A/G ratio levels were decreased and the decreased levels of mitochondrial enzymes were noted in CCl4-induced rats and the levels were restored near to normal significantly when the administration ethanolic extract of P. guajava, isolated quercetin fraction and silymarin. The normal architecture of liver tissues were altered and the mRNA expressions were up-regulated in CCl4-induced rats and the liver tissues were normalized and the mRNA and protein expressions were down-regulated near to normal significantly when the administration of ethanolic extract of P. guajava, isolated quercetin fraction and silymarin. CONCLUSION: From these results, the isolated quercetin fractions have better activity than that of the ethanolic extract of P. guajava leaves. Hence, the isolated quercetin may be used as the safest drug for hepatotoxicity in future.