Genome-Wide Identification and Analysis of Biotic and Abiotic Stress Regulation of C4 Photosynthetic Pathway Genes in Rice

Abstract

Photosynthetic fixation of CO₂ is more efficient in C₄ than in C₃ plants. Rice is a C₃ plant and a potential target for genetic engineering of the C₄ pathway. It is known that genes encoding C₄ enzymes are present in C₃ plants. However, no systematic analysis has been conducted to determine if these C₄ gene family members are expressed in diverse rice genotypes. In this study, we identified 15 genes belonging to the five C₄ gene families in rice genome through BLAST search using known maize C₄ photosynthetic pathway genes. Phylogenetic relationship of rice C₄ photosynthetic pathway genes and their isoforms with other grass genomes (Brachypodium, maize, Sorghum and Setaria), showed that these genes were highly conserved across grass genomes. Spatiotemporal, hormone, and abiotic stress specific expression pattern of the identified genes revealed constitutive as well as inductive responses of the C₄ photosynthetic pathway in different tissues and developmental stages of rice. Expression levels of C₄ specific gene family members in flag leaf during tillering stage were quantitatively analyzed in five rice genotypes covering three species, viz. Oryza sativa, ssp. japonica (cv. Nipponbare), Oryza sativa, ssp. indica (cv IR64, Swarna), and two wild species Oryza barthii and Oryza australiensis. The results showed that all the identified genes expressed in rice and exhibited differential expression pattern during different growth stages, and in response to biotic and abiotic stress conditions and hormone treatments. Our study concludes that C₄ photosynthetic pathway genes present in rice play a crucial role in stress regulation and might act as targets for C₄ pathway engineering via CRISPR-mediated breeding.

Keywords: C4 rice; Carbonic anhydrase (CA); Malate dehydrogenase (MDH); Malic enzyme (ME) and Pyruvate; Orthophosphate dikinase (PPDK); Phosphoenol pyruvate carboxylase (PEPC).