N-acetylglucosamine-1-P uridylyltransferase 1 and 2 are required for gametogenesis and embryo development in Arabidopsis thaliana.

ABSTRACT

Although N-acetylglucosamine-1-P uridylyltransferase (GlcNAc1pUT) that catalyzes the final step of the hexosamine biosynthetic pathway and is conserved among, organisms, produces UDP-N-acetylglucosamine (UDP-GlcNAc), an essential sugar moiety involved in protein glycosylation and structural polymers, its biological function in plants remains unknown. In this study, two GlcNA.UT genes were characterized in Arabidopsis thaliana. The single mutants glcna.ut1 and glcna.ut2 revealed no obvious phenotype, but their homozygous double mutant was lethal, reflecting the functional redundancy of these genes in being essential for plant growth. Mutant plants, GlcNA.UT1/glcna.ut1 glcna.ut2/ glcna.ut2, obtained from an F2-segregating population following reciprocal crosses of glcna.ut1 with glcna.ut2, displayed shorter siliques and fewer seed sets combined with impaired pollen viability and unfertilized ovules. Genetic analyses further demonstrated that the progeny of the GlcNA.UT1/glcna.ut1 glcna.ut2/glcna.ut2 mutant plants, but not those of the glcna.ut1/glcna.ut1 GlcNA.UT2/glcna.ut2 mutant plants, suffer from the aberrant transmission of (glcna.ut1 glcna.ut2) gametes. In parallel, cell biology analyses revealed a substantial defect in male gametophytes appearing during the late vacuolated or pollen mitosis I stages and that the female gametophyte is arrested during the uninucleate embryo sac stage in GlcNA.UT1/glcna.ut1 glcna.ut2/glcna.ut2 mutant plants. Nevertheless, although the glcna.ut1/glcna.ut1 GlcNA.UT2/glcna.ut2 mutant plants exhibited a normal transmission of (glcna.ut1 glcna.ut2) gametes and gametophytic development, the development of numerous embryos was arrested during the early globular stage within the embryo sacs. Collectively, despite having overlapping functions, the GlcNA.UT genes play an indispensable role in the unique mediation of gametogenesis and embryogenesis.