Human utilization of the mulberry–silkworm interaction started at least 5,000 years ago and greatly influenced world history through the Silk Road. Complementing the silkworm genome sequence, here we describe the genome of a mulberry species Morus notabilis. In the 330-Mb genome assembly, we identify 128 Mb of repetitive sequences and 29,338 genes, 60.8% of which are supported by transcriptome sequencing. Mulberry gene sequences appear to evolve ~3 times faster than other Rosales, perhaps facilitating the species’ spread worldwide. The mulberry tree is among a few eudicots but several Rosales that have not preserved genome duplications in more than 100 million years; however, a neopolyploid series found in the mulberry tree and several others suggest that new duplications may confer benefits. Five predicted mulberry miRNAs are found in the haemolymph and silk glands of the silkworm, suggesting interactions at molecular levels in the plant–herbivore relationship. The identification and analyses of mulberry genes involved in diversifying selection, resistance and protease inhibitor expressed in the laticifers will accelerate the improvement of mulberry plants.

MiRNAs are a class of non-coding endogenous small RNAs. They play vital roles in plant growth, development, and response to biotic and abiotic stress by negatively regulating genes. Mulberry trees are economically important species with multiple uses. However, to date, little is known about mulberry miRNAs and their target genes. In the present study, three small mulberry RNA libraries were constructed and sequenced using high-throughput sequencing technology. Results showed 85 conserved miRNAs belonging to 31 miRNA families and 262 novel miRNAs at 371 loci. Quantitative real-time PCR (qRT-PCR) analysis confirmed the expression pattern of 9 conserved and 5 novel miRNAs in leaves, bark, and male flowers. A total of 332 potential target genes were predicted to be associated with these 113 novel miRNAs. These results provide a basis for further understanding of mulberry miRNAs and the biological processes in which they are involved.

Mulberry is a fundamental component of the global sericulture industry, and its positive impact on our health and the environment cannot be overstated. However, the mulberry reference genomes reported previously remained unassembled or unplaced sequences. Here, we report the assembly and analysis of the telomere-to-telomere gap-free reference genome of the mulberry species, Morus notabilis, which has emerged as an important reference in mulberry gene function research and genetic improvement. The mulberry gap-free reference genome produced here provides an unprecedented opportunity for us to study the structure and function of centromeres. Our results revealed that all mulberry centromeric regions share conserved centromeric satellite repeats with different copies. Strikingly, we found that M. notabilis is a species with polycentric chromosomes and the only reported polycentric chromosome species up to now. We propose a compelling model that explains the formation mechanism of new centromeres and addresses the unsolved scientific question of the chromosome fusion-fission cycle in mulberry species. Our study sheds light on the functional genomics, chromosome evolution, and genetic improvement of mulberry species.

Proanthocyanidins (PAs) are major defense-related phenolics in mulberry, but the mechanism underlying their biosynthesis remains uncharacterized. In this study, the relationship between the expression of genes encoding anthocyanidin reductase (ANR) or leucoanthocyanidin reductase (LAR) and PA biosynthesis was investigated in white and red mulberry fruits. In ripening fruits, the MnANR and MnLAR transcription levels tended to decrease, whereas the catechin and epicatechin contents initially increased and then decreased. In contrast, the PA content exhibited a clearly different trend. The ectopic expression of MnANR and MnLAR in tobacco increased the resistance to Botrytis cinerea, as evidenced by the less extensive disease symptoms of the transgenic plants compared with the wild-type plants. In vitro experiments revealed that the transgenic tobacco crude leaf extract had an obvious inhibitory effect on B. cinerea. Moreover, the ectopic expression of MnANR and MnLAR in tobacco inhibited the expression of anthocyanin biosynthesis genes, resulting in decreased anthocyanin contents in flowers. The results of this study may be useful for elucidating the mechanism underlying PA biosynthesis. Furthermore, ANR and LAR represent potential targets for improving the resistance of mulberry and related plant species to B. cinerea.