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Development of an in vitro transformation system for Lupinus luteus

Li, H., Hoffman, K., Wylie, S.ORCID: 0000-0002-5639-7460, Ryan, K., Liu, L. and Jones, M.G.K.ORCID: 0000-0001-5002-0227 (1998) Development of an in vitro transformation system for Lupinus luteus. In Vitro Cellular & Developmental Biology - Animal, 34 (3). 53A.

Abstract

Lupins are a major crop in Western Australia, grown on 2.4 million acres of sandy soils in r~edium to nigh rainfall areas in the wheatbelt region. The major species grown is narrow-leafed lupin (Lupinus angustifolius). A further 1 million acres of poorly drained silty soils with low pH and a high aluminium content (Wodjil soils) exist on the eastern side of the wheatbelt where narrow-leafed lupins grow poorly. However, yellow lupins (L. luteus) tolerate such soils and one cultivar 'Wodjil', was released to growers in the 1997 season. 'Wodjil' is resistant to cucumber mosaic cucumovirus but susceptible to bean yellow mosaic potyvirus (BYMV), an important virus in the region. As genes for resistance to BYMV have not been detected in the yellow lupin germplasm, genetic engineering was seen as a method for introducing such genes, as well as other domestication traits. An in vitro transformation and regeneration system was developed for this species. Genes for herbicide resistance (bar) and BYMV resistance (Nla gene from BYMV) were introduced via Agrobacterium to hypocotyl segments which were grown in vitro until the shoots were big enough to graft to nontransgenic lupin rootstocks. Approximately 10% of explants produced roots /n vitro upon treatment with IBA. Grafted or rooted explants were transferred to hydroponic medium in the glasshouse. Seeds were harvested and grown on in soil. Transgenic plants were analysed by a number of methods including PCR, Southern analysis, bromocresol purple assay and PAT assay. T4 generation plants were subjected to BYMV challenge through manual and aphid inoculation and ELISA and visual observation indicated that resistance or delayed virus accumulation occurred in two transgenic lines.

Item Type: Journal Article
Murdoch Affiliation: Western Australian State Agricultural Biotechnology Centre
Publisher: Springer Verlag
Copyright: © 1998 Society for In Vitro Biology
Publishers Website: https://link.springer.com/article/10.1007/s11626-9...
Other Information: Abstract: 1998 Meeting of the Society for In Vitro Biology. Las Vegas, Nevada, 30 May - June 4 1998.
URI: http://researchrepository.murdoch.edu.au/id/eprint/49487
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