Single candidate gene for salt tolerance of Vigna nakashimae (Ohwi) Ohwi & Ohashi identified by QTL mapping, whole genome sequencing and triplicated RNA-seq analyses

• Published in: Breeding Science Vol. 74; no. 2; pp. 93 - 102
• Main Authors: Ito, Miho, Ohashi, Honami, Takemoto, Masahiro, Muto, Chiaki, Seiko, Takashi, Noda, Yusaku, Ogiso-Tanaka, Eri, Nagano, Atsushi J., Takahashi, Yu, Furukawa, Jun, Monden, Yuki, Naito, Ken
• Format: Journal Article
• Language: English
• Published: Japan Japanese Society of Breeding 01.01.2024; Japan Science and Technology Agency
• Subjects: Comparative analysis; Gene mapping; Gene sequencing; Genes; Genetic analysis; Genetic diversity; genetic resources; Genomes; Genomic analysis; Genotypes; genus Vigna; Groundwater; Groundwater depletion; Growth chambers; Nucleotide sequence; QTL; Quantitative trait loci; Research Paper; Ribonucleic acid; RNA; Salinity tolerance; Salinization; Salt tolerance; Soil analysis; Soil salinity; transcriptome analysis; Transcriptomes; Vigna nakashimae; Whole genome sequencing; wild crop relatives; whole genome sequencing; genus Vigna; QTL; salt tolerance; genetic resources; wild crop relatives; transcriptome analysis
• ISSN: 1344-7610; 1347-3735
• DOI: 10.1270/jsbbs.23053

Salt tolerance has been an important issue as a solution for soil salinization and groundwater depletion. To challenge this issue, genetic diversity of wild plants must be harnessed. Here we report a discovery of a candidate gene for salt tolerance in Vigna nakashimae, one of the coastal species in the genus Vigna. Using intraspecific variation, we performed a forward genetic analysis and identified a strong QTL region harboring ~200 genes. To further narrow down the candidate genes, we performed a comparative transcriptome analysis, using the genome sequence of azuki bean (V. angularis) as a reference. However the detected differentially-expressed genes (DEGs) did not include those related to salt tolerance. As we suspected that the target gene in V. nakashimae is missing in V. angularis, we sequenced the whole genome sequence of V. nakashimae with long-reads. By re-analyzing the transcriptome data with the new reference genome, we successfully identified POCO1 as a candidate gene, which was missing not only in V. angularis but also in the salt-sensitive accession of V. nakashimae. Further comparative analysis revealed that the tolerant genotypes conserved the ancestral form of the locus, while the sensitive genotypes did not. We also emphasize the pitfalls in our study, such as position effect in a growth chamber, missing important genes in the reference genome, and limited reproducibility of RNA-seq experiments.