Characterization of the ABC Transporter G Subfamily in Pomegranate and Function Analysis of PgrABCG14.

ATP-binding cassette subfamily G (ABCG) proteins play important roles in plant growth and development by transporting metabolites across cell membranes. To date, the genetic characteristics and potential functions of pomegranate ABCG proteins (PgrABCGs) have remained largely unknown. In this study, we found that 47 PgrABCGs were divided into five groups according to a phylogenetic analysis; groups I, II, III, and IV members are half-size proteins, and group V members are full-size proteins. PgrABCG14, PgrABCG21, and PgrABCG47 were highly expressed in the inner seed coat but had very low expression levels in the outer seed coat, and the expression levels of these three PgrABCG genes in the inner seed coats of hard-seeded pomegranate 'Dabenzi' were higher than those of soft-seeded pomegranate 'Tunisia'. In addition, the expression of these three PgrABCG genes was highly correlated with the expression of genes involved in lignin biosynthesis and hormone signaling pathways. The evolution of PgrABCG14 presents a highly similar trend to the origin and evolution of lignin biosynthesis during land plant evolution. Ectopic expression of PgrABCG14 in Arabidopsis promoted plant growth and lignin accumulation compared to wild type plants; meanwhile, the expression levels of lignin biosynthesis-related genes (CAD5, C4H, and Prx71) and cytokinin response marker genes (ARR5 and ARR15) were significantly upregulated in transgenic plants, which suggests the potential role of PgrABCG14 in promoting plant growth and lignin accumulation. Taken together, these findings not only provide insight into the characteristics and evolution of PgrABCGs, but also shed a light on the potential functions of PgrABCGs in seed hardness development.

Identification of sugar transporter (SWEET) genes involved in pomegranate seed coat sugar accumulation.

Sugar content of the outer seed coat and hardness of the inner seed coat are important traits of the pomegranate fruit. The translocation of sugars across biological membranes, mediated by SWEET transporters, is critical to seed development. In this study, we identified 16 PgrSWEET genes distributed on six chromosomes in the pomegranate genome. According to the phylogenetic analysis, PgrSWEET proteins were divided into four groups. Tandem and segmental duplications contributed to the expansion of the PgrSWEET family, while functional redundancy and diversification may have occurred among SWEET members according to analyses of evolution and gene expression. RNA-seq and qRT-PCR analyses revealed that PgrSWEET1a and PgrSWEET9 were highly expressed in the inner seed coat, and the expression levels gradually increased during seed development. Moreover, the relative expression levels of PgrSWEET1a and PgrSWEET9 in a hard-seeded cultivar were higher than those in a soft-seeded cultivar, indicating that PgrSWEET1a and PgrSWEET9 might function in the inner seed coat development by accumulating sugar metabolites. We also found that PgrSWEET2 was highly expressed in the outer seed coat during seed development, and the protein was localized to the tonoplast, indicating that PgrSWEET2 is likely a candidate regulating sugar accumulation or reutilization in the vacuoles of the outer seed coat. Genes encoding transcription factors probably regulating the candidate PgrSWEET genes were chosen by co-expression analysis. These results not only helped to characterize PgrSWEET genes but also provided an insight into their functions in relation to seed coat development. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03248-6.

Physiological adjustment of pomegranate pericarp responding to sunburn and its underlying molecular mechanisms.

BACKGROUND: Sunburn is common in pomegranate, and sunburned fruits have poor appearance and low marketability. However, the physiological and metabolic responses to sunburn and their underlying molecular mechanisms in pomegranate fruit are little understood. Fruit of sunburn-sensitive cultivar 'Hongyushizi' was used to carry out physiological parameter detection and widely-targeted metabolomics and transcriptome study. RESULTS: Malondialdehyde and relative conductivity increased with the severity of sunburn, which indicated increased membrane injury. Meanwhile, the content of antioxidants (total phenols and flavonoids), which reduce and repair membrane damage, increased and were accompanied by increases in total antioxidant capacity. In sunburned fruits compared with controls, 129 metabolites changed (including naringenin, pelargonidin and kaempferol) and 447 differentially expressed genes including CHI (Pgr25966.1), F3'5'H (Pgr26644.1), and CHS (Pgr005566.1) may have contributed to these changes. Transcription factors, such as NAC 5 (Pgr008725.1), MYB 93 (Pgr001791.1), and MYB 111 (Pgr027973.1) may be involved in phenylpropanoid and flavonoid biosynthesis by regulating the CHI, F3'5'H, and CHS etc. CONCLUSIONS: These findings provide insight into the sunburn mechanisms of pomegranate, and also into the genetic improvement of fruit sunburn.

Identification of Candidate Auxin Response Factors Involved in Pomegranate Seed Coat Development.

Auxin response factors (ARFs) are transcription factors, regulating the auxin signaling pathways involved in plant development and related processes. In this study, we performed the genome-wide identification and characterization of ARFs in pomegranate and compared them with ARFs from three other species. Seventeen PgrARFs were identified and clustered into four groups, according to their phylogenetic relationship with the remaining 59 ARFs. A recent whole-genome duplication event in pomegranate may have contributed to the expansion and diversification of PgrARFs. Genomic truncation and variant splicing mechanisms contributed to the divergence of PgrARFs, a conclusion that was supported by different exon-intron structures of genes and incomplete conserved domains of PgrARFs in a specific phylogenetic group (group III). Interestingly, the absence of motifs from certain PgrARF genes corresponded to their low transcription levels, which contrasted to the highly expressed PgrARFs with intact motifs. Specifically, PgrARF1 and PgrARF2 highly expressed in both inner and outer seed coat, and phylogenetically related to Arabidopsis orthologs which mediates cell divisions in seed coat. We infer these two PgrARFs might involve in seed coat development through cell divisions in response to auxin regulation. These findings provided information on the characteristics and evolutionary relationships of PgrARFs, but also shed lights on their potential roles during seed coat development in pomegranate.