A Carbonic Anhydrase, ZmCA4, Contributes to Photosynthetic Efficiency and Modulates CO2 Signaling Gene Expression by Interacting with Aquaporin ZmPIP2;6 in Maize.

Carbonic anhydrase (CA) catalyzes the reversible CO2 hydration reaction that produces bicarbonate for phosphoenolpyruvate carboxylase (PEPC). This is the initial step for transmitting the CO2 signal in C4 photosynthesis. However, it remains unknown whether the maize (Zea mays L.) CA gene, ZmCA4, plays a role in the maize photosynthesis process. In our study, we found that ZmCA4 was relatively highly expressed in leaves and localized in the chloroplast and the plasma membrane of mesophyll protoplasts. Knock-out of ZmCA4 reduced CA activity, while overexpression of ZmCA4 increased rubisco activity, as well as the quantum yield and relative electron transport rate in photosystem II. Overexpression of ZmCA4 enhanced maize yield-related traits. Moreover, ZmCA4 interacted with aquaporin ZmPIP2;6 in bimolecular fluorescence complementation and co-immunoprecipitation experiments. The double-knock-out mutant for ZmPIP2;6 and ZmCA4 genes showed reductions in its growth, CA and PEPC activities, assimilation rate and photosystem activity. RNA-Seq analysis revealed that the expression of other ZmCAs, ZmPIPs, as well as CO2 signaling pathway homologous genes, and photosynthetic-related genes was all altered in the double-knock-out mutant compared with the wild type. Altogether, our study's findings point to a critical role of ZmCA4 in determining photosynthetic capacity and modulating CO2 signaling regulation via its interaction with ZmPIP2;6, thus providing insight into the potential genetic value of ZmCA4 for maize yield improvement.

Genome-wide investigation of the LARP gene family: focus on functional identification and transcriptome profiling of ZmLARP6c1 in maize pollen.

BACKGROUND: The La-related proteins (LARPs) are a superfamily of RNA-binding proteins associated with regulation of gene expression. Evidence points to an important role for post-transcriptional control of gene expression in germinating pollen tubes, which could be aided by RNA-binding proteins. RESULTS: In this study, a genome-wide investigation of the LARP proteins in eight plant species was performed. The LARP proteins were classified into three families based on a phylogenetic analysis. The gene structure, conserved motifs, cis-acting elements in the promoter, and gene expression profiles were investigated to provide a comprehensive overview of the evolutionary history and potential functions of ZmLARP genes in maize. Moreover, ZmLARP6c1 was specifically expressed in pollen and ZmLARP6c1 was localized to the nucleus and cytoplasm in maize protoplasts. Overexpression of ZmLARP6c1 enhanced the percentage pollen germination compared with that of wild-type pollen. In addition, transcriptome profiling analysis revealed that differentially expressed genes included PABP homologous genes and genes involved in jasmonic acid and abscisic acid biosynthesis, metabolism, signaling pathways and response in a Zmlarp6c1::Ds mutant and ZmLARP6c1-overexpression line compared with the corresponding wild type. CONCLUSIONS: The findings provide a basis for further evolutionary and functional analyses, and provide insight into the critical regulatory function of ZmLARP6c1 in maize pollen germination.