Quinolizidine alkaloids are transported to seeds of bitter narrow-leafed lupin.

Narrow-leafed lupin (NLL, Lupinus angustifolius) is a promising legume crop that produces seeds with very high protein content. However, NLL accumulates toxic quinolizidine alkaloids (QAs) in most of its tissues, including the seeds. To determine the level of in situ biosynthesis in the seeds, we compared the accumulation of QAs with the expression of the biosynthetic gene lysine decarboxylase (LDC) in developing seeds and pods of a bitter (high-QA) variety of NLL. While QAs accumulated steadily in seeds until the drying phase, LDC expression was comparatively very low throughout seed development. In contrast, both QA accumulation and LDC expression peaked early in pods and decreased subsequently, reaching background levels at the onset of drying. We complemented these studies with MS imaging, which revealed the distribution patterns of individual QAs in cross-sections of pods and seeds. Finally, we show that a paternal bitter genotype does not influence the QA levels of F1 seeds grown on a maternal, low-QA genotype. We conclude that the accumulation of QAs in seeds of bitter NLL is mostly, if not exclusively, transported from other tissues. These results open the possibility of using transport engineering to generate herbivore-resistant bitter NLL varieties that produce QA-free seeds.

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

Lupins (Lupinus spp.) are nitrogen-fixing legumes that accumulate toxic alkaloids in their protein-rich beans. These anti-nutritional compounds belong to the family of quinolizidine alkaloids (QAs), which are of interest to the pharmaceutical and chemical industries. To unleash the potential of lupins as protein crops and as sources of QAs, a thorough understanding of the QA pathway is needed. However, only the first enzyme in the pathway, lysine decarboxylase (LDC), is known. Here, we report the transcriptome of a high-QA variety of narrow-leafed lupin (L. angustifolius), obtained using eight different tissues and two different sequencing technologies. In addition, we present a list of 33 genes that are closely co-expressed with LDC and that represent strong candidates for involvement in lupin alkaloid biosynthesis. One of these genes encodes a copper amine oxidase able to convert the product of LDC, cadaverine, into 1-piperideine, as shown by heterologous expression and enzyme assays. Kinetic analysis revealed a low KM value for cadaverine, supporting a role as the second enzyme in the QA pathway. Our transcriptomic data set represents a crucial step towards the discovery of enzymes, transporters, and regulators involved in lupin alkaloid biosynthesis.