Chlorophyll deficiency delays but does not prevent melanogenesis in barley seed melanoplasts

Plant melanin is a dark polymerized polyphenolic substance that can by synthesized in seed tissues. Unlike well-defined enzymatic browning reaction leading to melanin synthesis in senescent and damaged plant tissues, melanin formation in intact tissues was not studied properly. Recently, melanin synthesis was demonstrated in chloroplast-derived melanoplasts in pericarp and husk cells of barley seeds. In barley, there are two independent genes, Blp1 and Alm1, affecting respectively the biosynthesis of melanin and chlorophyll in seeds. Even though different genetic systems are responsible for these traits, the localization of these biosynthetic pathways in the same organelle prompted us to conduct an in-depth study of the i:Bwalm1Blp1 line characterized by simultaneous chlorophyll deficiency caused by recessive allele alm1 and melanin accumulation controlled by dominant allele Blp1. This barley line and parental ones-Bowman, i:BwBlp1, and i:Bwalm1, which are characterized by different combinations of pigments chlorophyll and melanin in seeds-were subjected to a comparative cytological analysis. Three markers were analyzed: the presence of visible pigments, chlorophyll, and PsbA protein (a thylakoid membrane marker). Plastids of the barley pericarp and husk showed prominent differences among the lines, with internal structures that are more developed in husk cells. Although chlorophyll deficiency did not prevent melanogenesis in the spike of the hybrid line, a 7-day delay in melanization initiation and a decrease in its magnitude were observed in comparison with the melanin-and-chlorophyll-containing line. Thus, melanin biosynthesis is not related to photosynthetic processes directly but may be dependent on the presence of plastids with well-developed internal membranes.