Anthocyanin Accumulation and Related Gene Expression Profile in ‘Red Zaosu’ Pear and Its Green Mutant

Red-skinned pear is a promising commercial fruit due to its attractive appearance and nutritious value. Anthocyanin is the determinant of the red coloration of the pear peel. However, differences in anthocyanin accumulation exist among red pear cultivars with different genetic backgrounds. In this study, we analyzed the anthocyanin content and gene expression patterns in the fruits and different tissues of the red pear ‘Red Zaosu’ at different developmental stages and found a difference in anthocyanin accumulation between ‘Red Zaosu’ pear and its green mutant. The data showed that the expression profiles of transcripts that encoded critical anthocyanin biosynthetic genes were basically consistent with a tendency to a decreased anthocyanin content during fruit development, indicating that a synergistic effect of these genes was responsible for anthocyanin biosynthesis and regulation. Tissue-specific expression analysis of anthocyanin biosynthetic genes showed that they could be expressed in all tissues but at different levels. PbF3H, PbDFR, and PbANS were mainly expressed during the early flowering period, which explained the reduced levels of anthocyanin content in petals. Additionally, the content of anthocyanins and the expression levels of PbDFR, PbANS, and PbMYB10 significantly decreased in the green mutant of ‘Red Zaosu’, suggesting that PbDFR, PbANS, and PbMYB10 probably play a decisive role in determining the skin coloration of ‘Red Zaosu’ and its green mutant.

An Argon-Ion-Induced Pale Green Mutant of Arabidopsis Exhibiting Rapid Disassembly of Mesophyll Chloroplast Grana

Argon-ion beam is an effective mutagen capable of inducing a variety of mutation types. In this study, an argon ion-induced pale green mutant of Arabidopsis thaliana was isolated and characterized. The mutant, designated Ar50-33-pg1, exhibited moderate defects of growth and greening and exhibited rapid chlorosis in photosynthetic tissues. Fluorescence microscopy confirmed that mesophyll chloroplasts underwent substantial shrinkage during the chlorotic process. Genetic and whole-genome resequencing analyses revealed that Ar50-33-pg1 contained a large 940 kb deletion in chromosome V that encompassed more than 100 annotated genes, including 41 protein-coding genes such as TYRAAt1/TyrA1, EGY1, and MBD12. One of the deleted genes, EGY1, for a thylakoid membrane-localized metalloprotease, was the major contributory gene responsible for the pale mutant phenotype. Both an egy1 mutant and F1 progeny of an Ar50-33-pg1 × egy1 cross-exhibited chlorotic phenotypes similar to those of Ar50-33-pg1. Furthermore, ultrastructural analysis of mesophyll cells revealed that Ar50-33-pg1 and egy1 initially developed wild type-like chloroplasts, but these were rapidly disassembled, resulting in thylakoid disorganization and fragmentation, as well as plastoglobule accumulation, as terminal phenotypes. Together, these data support the utility of heavy-ion mutagenesis for plant genetic analysis and highlight the importance of EGY1 in the structural maintenance of grana in mesophyll chloroplasts.

Defect in Brnym1, a magnesium-dechelatase protein, causes a stay-green phenotype in an EMS-mutagenized Chinese cabbage (Brassica campestris L. ssp. pekinensis) line

Leaf color is an important target trait in Chinese cabbage breeding programs. Leaf yellowing may reduce crop commercial and nutritional values. Some plants with the “stay-green” trait maintain leaf greenness during senescence and even after death. Stay-green Chinese cabbage may be a focal point of future breeding projects because it could improve crop quality and yield and prolong shelf life. A new stay-green mutant, non-yellowing mutant 1 (nym1), was identified in Chinese cabbage derived from an ethyl methane sulfonate (EMS)-mutagenized population. The mutant had stay-green characteristics and a higher chlorophyll content than the wild-type during leaf senescence. The stay-green trait in the mutant Chinese cabbage was controlled by the recessive gene Brnym1. MutMap and KASP analyses showed that Brnym1 (BraA03g050600.3C) encodes an mg-dechelatase (SGR protein), which might be the causal gene of the mutation in Chinese cabbage. A nonsynonymous single nucleotide base substitution (G to A) in the third exon of Brnym1 caused an amino acid substitution from L to F in the highly conserved domain of the magnesium-dechelatase. Ectopic overexpression showed that the BrNYM1 gene of wild-type Chinese cabbage complemented the SGR-defective stay-green mutant nye1-1 of Arabidopsis. The magnesium-dechelatase activity in the nym1 mutant was significantly downregulated compared to that in the wild type. Brnym1 was relatively upregulated in the mutant during late senescence, and BrNYM1 was localized to the chloroplasts. These results indicate that Brnym1 (BraA03g050600.3C) is the causal gene of the stay-green mutation and could be of particular significance in the genetic improvement of Chinese cabbage.