Functional loss of pAMT results in biosynthesis of capsinoids, capsaicinoid analogs, in capsicum annuum cv. CH-19 sweet

Yaqin Lang, Hiroaki Kisaka, Ryuji Sugiyama, Kenzo Nomura, Akihito Morita, Tatsuo Watanabe, Yoshiyuki Tanaka, Susumu Yazawa, Tetsuya Miwa

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

Capsaicinoids are responsible for the spicy flavor of pungent peppers (Capsicum). The cultivar CH-19 Sweet is a non-pungent pepper mutant derived from a pungent pepper strain, Capsicum annuum CH-19. CH-19 Sweet biosynthesizes capsaicinoid analogs, capsinoids. We determined the genetic and metabolic mechanisms of capsinoid biosynthesis in this cultivar. We analyzed the putative aminotransferase (pAMT) that is thought to catalyze the formation of vanillylamine from vanillin in the capsaicinoid biosynthetic pathway. Enzyme assays revealed that pAMT activity catalyzing vanillylamine formation was completely lost in CH-19 Sweet placenta tissue. RT-PCR analysis showed normal mRNA transcription of the pAMT gene; however, SNP analysis of the cDNA sequence showed a T nucleotide insertion at 1291 bp in the pAMT gene of CH-19 Sweet. This insertion formed a new stop codon, TGA, that prevented normal translation of the gene, and the pAMT protein did not accumulate in CH-19 Sweet as determined using Western blot analysis. We developed a dCAPS marker based on the T insertion in the pAMT gene of CH-19 Sweet, and showed that the pAMT genotype co-segregated with the capsinoid or capsaicinoid fruit phenotype in the F 2 population. The T insertion was not found in other pungent and non-pungent Capsicum lines, suggesting that it is specific to CH-19 Sweet. CH-19 Sweet's pAMT gene mutation is an example of a nonsense mutation in a single gene that alters a secondary metabolite biosynthetic pathway, resulting in the biosynthesis of analogs. The dCAPS marker will be useful in selecting lines with capsinoid-containing fruits in pepper-breeding programs. Journal compilation

Original languageEnglish
Pages (from-to)953-961
Number of pages9
JournalPlant Journal
Volume59
Issue number6
DOIs
Publication statusPublished - Sep 2009
Externally publishedYes

Fingerprint

Capsicum
transaminases
Capsicum annuum
Transaminases
biosynthesis
sweet peppers
Genes
genes
Biosynthetic Pathways
biochemical pathways
Fruit
nonsense mutation
fruits
Terminator Codon
vanillin
stop codon
Nonsense Codon
Enzyme Assays
cultivars
placenta

Keywords

  • Capsicum
  • Capsinoids
  • Putative aminotransferase (pAMT)
  • T nucleotide insertion
  • Vanillyl alcohol
  • Vanillylamine

ASJC Scopus subject areas

  • Plant Science
  • Cell Biology
  • Genetics

Cite this

Functional loss of pAMT results in biosynthesis of capsinoids, capsaicinoid analogs, in capsicum annuum cv. CH-19 sweet. / Lang, Yaqin; Kisaka, Hiroaki; Sugiyama, Ryuji; Nomura, Kenzo; Morita, Akihito; Watanabe, Tatsuo; Tanaka, Yoshiyuki; Yazawa, Susumu; Miwa, Tetsuya.

In: Plant Journal, Vol. 59, No. 6, 09.2009, p. 953-961.

Research output: Contribution to journalArticle

Lang, Y, Kisaka, H, Sugiyama, R, Nomura, K, Morita, A, Watanabe, T, Tanaka, Y, Yazawa, S & Miwa, T 2009, 'Functional loss of pAMT results in biosynthesis of capsinoids, capsaicinoid analogs, in capsicum annuum cv. CH-19 sweet', Plant Journal, vol. 59, no. 6, pp. 953-961. https://doi.org/10.1111/j.1365-313X.2009.03921.x
Lang, Yaqin ; Kisaka, Hiroaki ; Sugiyama, Ryuji ; Nomura, Kenzo ; Morita, Akihito ; Watanabe, Tatsuo ; Tanaka, Yoshiyuki ; Yazawa, Susumu ; Miwa, Tetsuya. / Functional loss of pAMT results in biosynthesis of capsinoids, capsaicinoid analogs, in capsicum annuum cv. CH-19 sweet. In: Plant Journal. 2009 ; Vol. 59, No. 6. pp. 953-961.
@article{3fdc12bd8077403c9e94f4630650cc8b,
title = "Functional loss of pAMT results in biosynthesis of capsinoids, capsaicinoid analogs, in capsicum annuum cv. CH-19 sweet",
abstract = "Capsaicinoids are responsible for the spicy flavor of pungent peppers (Capsicum). The cultivar CH-19 Sweet is a non-pungent pepper mutant derived from a pungent pepper strain, Capsicum annuum CH-19. CH-19 Sweet biosynthesizes capsaicinoid analogs, capsinoids. We determined the genetic and metabolic mechanisms of capsinoid biosynthesis in this cultivar. We analyzed the putative aminotransferase (pAMT) that is thought to catalyze the formation of vanillylamine from vanillin in the capsaicinoid biosynthetic pathway. Enzyme assays revealed that pAMT activity catalyzing vanillylamine formation was completely lost in CH-19 Sweet placenta tissue. RT-PCR analysis showed normal mRNA transcription of the pAMT gene; however, SNP analysis of the cDNA sequence showed a T nucleotide insertion at 1291 bp in the pAMT gene of CH-19 Sweet. This insertion formed a new stop codon, TGA, that prevented normal translation of the gene, and the pAMT protein did not accumulate in CH-19 Sweet as determined using Western blot analysis. We developed a dCAPS marker based on the T insertion in the pAMT gene of CH-19 Sweet, and showed that the pAMT genotype co-segregated with the capsinoid or capsaicinoid fruit phenotype in the F 2 population. The T insertion was not found in other pungent and non-pungent Capsicum lines, suggesting that it is specific to CH-19 Sweet. CH-19 Sweet's pAMT gene mutation is an example of a nonsense mutation in a single gene that alters a secondary metabolite biosynthetic pathway, resulting in the biosynthesis of analogs. The dCAPS marker will be useful in selecting lines with capsinoid-containing fruits in pepper-breeding programs. Journal compilation",
keywords = "Capsicum, Capsinoids, Putative aminotransferase (pAMT), T nucleotide insertion, Vanillyl alcohol, Vanillylamine",
author = "Yaqin Lang and Hiroaki Kisaka and Ryuji Sugiyama and Kenzo Nomura and Akihito Morita and Tatsuo Watanabe and Yoshiyuki Tanaka and Susumu Yazawa and Tetsuya Miwa",
year = "2009",
month = "9",
doi = "10.1111/j.1365-313X.2009.03921.x",
language = "English",
volume = "59",
pages = "953--961",
journal = "Plant Journal",
issn = "0960-7412",
publisher = "Wiley-Blackwell",
number = "6",

}

TY - JOUR

T1 - Functional loss of pAMT results in biosynthesis of capsinoids, capsaicinoid analogs, in capsicum annuum cv. CH-19 sweet

AU - Lang, Yaqin

AU - Kisaka, Hiroaki

AU - Sugiyama, Ryuji

AU - Nomura, Kenzo

AU - Morita, Akihito

AU - Watanabe, Tatsuo

AU - Tanaka, Yoshiyuki

AU - Yazawa, Susumu

AU - Miwa, Tetsuya

PY - 2009/9

Y1 - 2009/9

N2 - Capsaicinoids are responsible for the spicy flavor of pungent peppers (Capsicum). The cultivar CH-19 Sweet is a non-pungent pepper mutant derived from a pungent pepper strain, Capsicum annuum CH-19. CH-19 Sweet biosynthesizes capsaicinoid analogs, capsinoids. We determined the genetic and metabolic mechanisms of capsinoid biosynthesis in this cultivar. We analyzed the putative aminotransferase (pAMT) that is thought to catalyze the formation of vanillylamine from vanillin in the capsaicinoid biosynthetic pathway. Enzyme assays revealed that pAMT activity catalyzing vanillylamine formation was completely lost in CH-19 Sweet placenta tissue. RT-PCR analysis showed normal mRNA transcription of the pAMT gene; however, SNP analysis of the cDNA sequence showed a T nucleotide insertion at 1291 bp in the pAMT gene of CH-19 Sweet. This insertion formed a new stop codon, TGA, that prevented normal translation of the gene, and the pAMT protein did not accumulate in CH-19 Sweet as determined using Western blot analysis. We developed a dCAPS marker based on the T insertion in the pAMT gene of CH-19 Sweet, and showed that the pAMT genotype co-segregated with the capsinoid or capsaicinoid fruit phenotype in the F 2 population. The T insertion was not found in other pungent and non-pungent Capsicum lines, suggesting that it is specific to CH-19 Sweet. CH-19 Sweet's pAMT gene mutation is an example of a nonsense mutation in a single gene that alters a secondary metabolite biosynthetic pathway, resulting in the biosynthesis of analogs. The dCAPS marker will be useful in selecting lines with capsinoid-containing fruits in pepper-breeding programs. Journal compilation

AB - Capsaicinoids are responsible for the spicy flavor of pungent peppers (Capsicum). The cultivar CH-19 Sweet is a non-pungent pepper mutant derived from a pungent pepper strain, Capsicum annuum CH-19. CH-19 Sweet biosynthesizes capsaicinoid analogs, capsinoids. We determined the genetic and metabolic mechanisms of capsinoid biosynthesis in this cultivar. We analyzed the putative aminotransferase (pAMT) that is thought to catalyze the formation of vanillylamine from vanillin in the capsaicinoid biosynthetic pathway. Enzyme assays revealed that pAMT activity catalyzing vanillylamine formation was completely lost in CH-19 Sweet placenta tissue. RT-PCR analysis showed normal mRNA transcription of the pAMT gene; however, SNP analysis of the cDNA sequence showed a T nucleotide insertion at 1291 bp in the pAMT gene of CH-19 Sweet. This insertion formed a new stop codon, TGA, that prevented normal translation of the gene, and the pAMT protein did not accumulate in CH-19 Sweet as determined using Western blot analysis. We developed a dCAPS marker based on the T insertion in the pAMT gene of CH-19 Sweet, and showed that the pAMT genotype co-segregated with the capsinoid or capsaicinoid fruit phenotype in the F 2 population. The T insertion was not found in other pungent and non-pungent Capsicum lines, suggesting that it is specific to CH-19 Sweet. CH-19 Sweet's pAMT gene mutation is an example of a nonsense mutation in a single gene that alters a secondary metabolite biosynthetic pathway, resulting in the biosynthesis of analogs. The dCAPS marker will be useful in selecting lines with capsinoid-containing fruits in pepper-breeding programs. Journal compilation

KW - Capsicum

KW - Capsinoids

KW - Putative aminotransferase (pAMT)

KW - T nucleotide insertion

KW - Vanillyl alcohol

KW - Vanillylamine

UR - http://www.scopus.com/inward/record.url?scp=70349139708&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=70349139708&partnerID=8YFLogxK

U2 - 10.1111/j.1365-313X.2009.03921.x

DO - 10.1111/j.1365-313X.2009.03921.x

M3 - Article

VL - 59

SP - 953

EP - 961

JO - Plant Journal

JF - Plant Journal

SN - 0960-7412

IS - 6

ER -