Transcript profiling of a bitter variety of narrow-leafed lupin to discover alkaloid biosynthetic genes

Yang, Ting; Nagy, István; Mancinotti, Davide; Otterbach, Sophie Lisa; Andersen, Trine Bundgaard; Motawia, M. S.; Asp, Torben; Geu‐Flores, Fernando

doi:10.1093/jxb/erx362

Cited by 47 publications

(90 citation statements)

References 59 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Lup000530.1 was detected in the stem and leaf tissue of P27255 but not in any tissues of Tanjil and Unicrop (Figure S1) and represented a good candidate for involvement in the QA biosynthetic pathway. The enzymatic function of this gene was recently validated by Yang et al () and is designated LaCAO for follow‐up gene expression analysis in this study.…”

Section: Resultsmentioning

confidence: 81%

“…LaL/ODC is involved in the first step of the QA biosynthetic pathway, the decarboxylation of lysine to the intermediate cadaverine (Bunsupa et al, ). A copper amine oxidase functions in the deamination of cadaverine to yield a pathway intermediate common to the formation of all QAs (Golebiewski & Spenser, ; Leistner & Spenser, ), with LaCAO recently identified as being involved in this reaction (Yang et al, ). After the formation of QAs such as lupanine, sparteine, multiflorine, and lupinine, further modification can yield a variety of structurally related QAs (Ohmiya, Saito, & Murakoshi, ; Saito, Suzuki, Takamatsu, & Murakoshi, ; Suzuki, Murakoshi, & Saito, ; Wink & Hartmann, ).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Characterization of the genetic factors affecting quinolizidine alkaloid biosynthesis and its response to abiotic stress in narrow‐leafed lupin (Lupinus angustifoliusL.)

et al. 2018

View full text Add to dashboard Cite

Quinolizidine alkaloids (QAs) are toxic secondary metabolites that complicate the end use of narrow-leafed lupin (NLL; Lupinus angustifolius L.) grain, as levels sometimes exceed the industry limit for its use as a food and feed source. The genotypic and environmental influences on QA production in NLL are poorly understood. Here, the expression of QA biosynthetic genes was analysed in vegetative and reproductive tissues of bitter (high QA) and sweet (low QA) accessions. It was demonstrated that sweet accessions are characterized by lower QA biosynthetic gene expression exclusively in leaf and stem tissues than bitter NLL, consistent with the hypothesis that QAs are predominantly produced in aerial tissues and transported to seeds, rather than synthesized within the seed itself. This analysis informed our identification of additional candidate genes involved in QA biosynthesis. Drought and temperature stress are two major abiotic stresses that often occur during NLL pod set. Hence, we assessed the effect of drought, increased temperature, and their combination, on QA production in three sweet NLL cultivars. A cultivar-specific response to drought and temperature in grain QA levels was observed, including the identification of a cultivar where alkaloid levels did not change with these stress treatments.

show abstract

Section: Resultsmentioning

confidence: 81%

Section: Introductionmentioning

confidence: 99%

Characterization of the genetic factors affecting quinolizidine alkaloid biosynthesis and its response to abiotic stress in narrow‐leafed lupin (Lupinus angustifoliusL.)

et al. 2018

View full text Add to dashboard Cite

show abstract

“…Phylogenetic analysis using all eight CuAO or CuAO-like enzymes from Arabidopsis with other plant CuAOs resulted in the formation of three major clades ( Figure S15). Among the eight Arabidopsis CuAOs, AtCuAO3 was classified in clade III, which included four previously characterized enzymes from L. angustifolius (LaCuAO), N. tabacum (NtDAO1 and NtMPO) and Malus domestica (MdAO1), all of which have shown activity towards cadaverine (Naconsie et al, 2014;Zarei et al, 2015;Yang et al, 2017). Figure 5.…”

Section: Cadaverine Catabolic Pathway In Arabidopsis Emerged By Ectopmentioning

confidence: 99%

Metabolic diversification of nitrogen‐containing metabolites by the expression of a heterologous lysine decarboxylase gene in Arabidopsis

et al. 2019

View full text Add to dashboard Cite

Summary Lysine decarboxylase converts l‐lysine to cadaverine as a branching point for the biosynthesis of plant Lys‐derived alkaloids. Although cadaverine contributes towards the biosynthesis of Lys‐derived alkaloids, its catabolism, including metabolic intermediates and the enzymes involved, is not known. Here, we generated transgenic Arabidopsis lines by expressing an exogenous lysine/ornithine decarboxylase gene from Lupinus angustifolius (La‐L/ODC) and identified cadaverine‐derived metabolites as the products of the emerged biosynthetic pathway. Through untargeted metabolic profiling, we observed the upregulation of polyamine metabolism, phenylpropanoid biosynthesis and the biosynthesis of several Lys‐derived alkaloids in the transgenic lines. Moreover, we found several cadaverine‐derived metabolites specifically detected in the transgenic lines compared with the non‐transformed control. Among these, three specific metabolites were identified and confirmed as 5‐aminopentanal, 5‐aminopentanoate and δ‐valerolactam. Cadaverine catabolism in a representative transgenic line (DC29) was traced by feeding stable isotope‐labeled [α‐15N]‐ or [ε‐15N]‐l‐lysine. Our results show similar 15N incorporation ratios from both isotopomers for the specific metabolite features identified, indicating that these metabolites were synthesized via the symmetric structure of cadaverine. We propose biosynthetic pathways for the metabolites on the basis of metabolite chemistry and enzymes known or identified through catalyzing specific biochemical reactions in this study. Our study shows that this pool of enzymes with promiscuous activities is the driving force for metabolite diversification in plants. Thus, this study not only provides valuable information for understanding the catabolic mechanism of cadaverine but also demonstrates that cadaverine accumulation is one of the factors to expand plant chemodiversity, which may lead to the emergence of Lys‐derived alkaloid biosynthesis.

show abstract

“…Primers used in this study for quantitative real-time -PCR have been described previously (Frick et al 2018). These primers are used to amplify the quantitative real-time PCR reference gene L. angustifolius UBC9-like (LaUBC9-like; Lup018255.1), QA biosynthetic genes LaL/ODC (Lup009726.1) (Bunsupa et al 2012), LaCAO (Lup000530.1) (Yang et al 2017a) and LaHMT/ HLT (Lup022251.1) (Okada et al 2005) and candidate biosynthetic gene LaAT (Lup021586.1) (Bunsupa et al 2011).…”

Section: Primer Design and Quantitative Real-time -Pcrmentioning

confidence: 99%

“…We then measured the expression of four key QA biosynthetic genes in response to these treatments and compared this expression to the observed induction of the JA pathway. Although the QA biosynthetic pathway is only partly elucidated (Frick et al 2017), genes that have been molecularly characterised as having a role in QA biosynthesis include L. angustifolius lysine/ornithine decarboxylase (LaL/ODC), L. angustifolius copper amine oxidase (LaCAO) and L. albus tigloyl-CoA: (Okada et al 2005;Bunsupa et al 2012;Yang et al 2017a). L. angustifolius acyltransferase (LaAT) has also been suggested as being involved in QA biosynthesis (Bunsupa et al 2011).…”

Section: Introductionmentioning

confidence: 99%

The role of jasmonate signalling in quinolizidine alkaloid biosynthesis, wounding and aphid predation response in narrow-leafed lupin

Frick

Foley

Siddique

et al. 2019

Functional Plant Biol.

View full text Add to dashboard Cite

Quinolizidine alkaloids (QAs) are toxic secondary metabolites produced in lupin species that protect the plant against insects. They form in vegetative tissues and accumulate to a different extent in the grains: high levels in ‘bitter’ narrow-leafed lupin (NLL) and low levels in ‘sweet’ NLL. Grain QA levels vary considerably, and sometimes exceed the industry limit for food and feed purposes. We hypothesised that jasmonates regulate QA biosynthesis in response to environmental stresses such as wounding and aphid predation, which may explain non-genetic variability in grain QA levels. Methyl jasmonate (MeJA)-inducible genes were identified and verified in NLL. Exogenous MeJA application-induced expression of QA biosynthetic genes and QA levels for bitter, but not sweet NLL. Although MeJA-inducible genes responded to wounding, the expression of QA biosynthetic genes was not induced for bitter and sweet NLL. We assessed the effect of aphid predation on QA production for two cultivars – one moderately resistant and one susceptible to aphid predation. Although MeJA-inducible genes responded to aphid predation, no change in QA levels was found for either cultivar. These findings offer insights into the regulation of QA biosynthesis in bitter and sweet NLL and concludes that aphids are not a concern for increasing grain QAs in NLL cultivars.

show abstract

Transcript profiling of a bitter variety of narrow-leafed lupin to discover alkaloid biosynthetic genes

Abstract: We report the first transcript profiling of a high-alkaloid lupin variety and identify a new alkaloid biosynthetic gene, thus accelerating the efforts to develop alkaloid-free lupin crops.

Cited by 47 publications

References 59 publications

Characterization of the genetic factors affecting quinolizidine alkaloid biosynthesis and its response to abiotic stress in narrow‐leafed lupin (Lupinus angustifoliusL.)

Characterization of the genetic factors affecting quinolizidine alkaloid biosynthesis and its response to abiotic stress in narrow‐leafed lupin (Lupinus angustifoliusL.)

Metabolic diversification of nitrogen‐containing metabolites by the expression of a heterologous lysine decarboxylase gene in Arabidopsis

The role of jasmonate signalling in quinolizidine alkaloid biosynthesis, wounding and aphid predation response in narrow-leafed lupin

Contact Info

Product

Resources

About