Identification of a Novel Mutant pAMT Allele Responsible for Low-pungency and Capsinoid Production in Chili Pepper: Accession ‘No. 4034’ (Capsicum chinense)

Abstract

Capsinoids are low-pungent capsaicinoid analogues in chili pepper fruits. They exhibit various bioactivities in humans similar to capsaicinoids, but do not produce a nasty burning sensation, encouraging their application in foods and supplements. Previous reports demonstrated that loss-of-function of putative aminotransferase (pAMT) leads to low-pungency and capsinoid accumulation. Therefore, the pamt allele is a useful gene in chili pepper breeding programs to enhance health-promoting properties. Eight loss-of-function alleles have been identified in the Capsicum genus, but the variation in pamt alleles remains to be fully elucidated. In this study, we identified one novel loss-of-function allele from the analysis of low-pungent chili pepper ‘No. 4034’ (C. chinense). ‘No. 4034’ contained mainly capsinoid with an undetectable level of capsaicinoid. A genetic complementation test was conducted by crossing ‘No. 4034’ with other accessions. The results indicated that ‘No. 4034’ possessed a loss-of-function pamt allele. Sequence analysis showed that the novel mutant allele contained a 7-bp insertion (TCGGTAC) in the 16th exon region, which we designated as pamt⁹. The insertion caused a frameshift mutation and resulted in a truncated protein. Gene expression analysis showed that the expression level of pAMT specifically decreased among biosynthetic genes tested here in ‘No. 4034’, compared with that of pungent accession. pamt⁹ will be useful for low-pungency and capsinoid breeding, and will provide additional information for variations in pAMT mutants.