Regulation of the plastochron by three many-noded dwarf genes in barley

Author summary The number of leaves produced during a plant's lifetime is major determinant of plant architecture and affects the efficiency of photosynthesis and crop productivity. The leaf number is dependent on the temporal pattern of leaf initiation at the shoot apical meristem, which is termed the plastochron. The genetic factors involved in plastochron regulation have been identified in several plant species. However, whether the functions of plastochron-related genes and their genetic pathways are universal or diversified among different plant species is unclear. In this study, we investigated many-noded dwarf (mnd) mutants in barley, which exhibited a shortened plastochron and a decreased leaf blade length. The mutant alleles used in this study were derived from three genes, MND4, MND1, and MND8, which encode a cytochrome P450 family protein, an N-acetyltransferase-like protein, and a MATE transporter-family protein, respectively. Phenotypic and expression analyses revealed that these three MND genes affect the leaf production rate and leaf maturation program, but their expression levels were interdependent. In addition, the plastochron and leaf growth are closely related but independently regulated. We also analyzed the expression patterns and knockout mutants of rice MND orthologs to clarify whether their biological functions are conserved in rice and barley. This study provides insight into the genetic mechanisms of plastochron control in grass species. The plastochron, the time interval between the formation of two successive leaves, is an important determinant of plant architecture. We genetically and phenotypically investigated many-noded dwarf (mnd) mutants in barley. The mnd mutants exhibited a shortened plastochron and a decreased leaf blade length, and resembled previously reported plastochron1 (pla1), pla2, and pla3 mutants in rice. In addition, the maturation of mnd leaves was accelerated, similar to pla mutants in rice. Several barley mnd alleles were derived from three genes-MND1, MND4, and MND8. Although MND4 coincided with a cytochrome P450 family gene that is a homolog of rice PLA1, we clarified that MND1 and MND8 encode an N-acetyltransferase-like protein and a MATE transporter-family protein, which are respectively orthologs of rice GW6a and maize BIGE1 and unrelated to PLA2 or PLA3. Expression analyses of the three MND genes revealed that MND1 and MND4 were expressed in limited regions of the shoot apical meristem and leaf primordia, but MND8 did not exhibit a specific expression pattern around the shoot apex. In addition, the expression levels of the three genes were interdependent among the various mutant backgrounds. Genetic analyses using the double mutants mnd4mnd8 and mnd1mnd8 indicated that MND1 and MND4 regulate the plastochron independently of MND8, suggesting that the plastochron in barley is controlled by multiple genetic pathways involving MND1, MND4, and MND8. Correlation analysis between leaf number and leaf blade length indicated that both traits exhibited a strong negative association among different genetic backgrounds but not in the same genetic background. We propose that MND genes function in the regulation of the plastochron and leaf growth and revealed conserved and diverse aspects of plastochron regulation via comparative analysis of barley and rice.

Automated summary

This study investigated the genetic mechanisms of plastochron control in grass species using many-noded dwarf mutants in barley. The MND genes were found to play important roles in regulating leaf growth and plastochron independently in barley. The study revealed both conserved and diverse aspects of plastochron regulation between barley and rice.