Article

Molecular Breeding

, Volume 23, Issue 1, pp 153-161

First online:

Development of a sequence-specific PCR marker linked to the gene “pauper” conferring low-alkaloids in white lupin (Lupinus albus L.) for marker assisted selection

  • Ruiming LinAffiliated withInstitute of Plant Protection, Chinese Academy of Agricultural Sciences
  • , Daniel RenshawAffiliated withDepartment of Agriculture and Food Western Australia
  • , David LuckettAffiliated withEH Graham Centre for Agricultural Innovation, NSW Department of Primary Industries, Agricultural Institute
  • , Jonathon ClementsAffiliated withDepartment of Agriculture and Food Western AustraliaThe Centre for Legumes in Mediterranean Agriculture, The University of Western Australia
  • , Guijun YanAffiliated withSchool of Plant Biology, Faculty of Natural and Agricultural Sciences, The University of Western Australia
  • , Kedar AdhikariAffiliated withDepartment of Agriculture and Food Western Australia
  • , Bevan BuirchellAffiliated withDepartment of Agriculture and Food Western Australia
  • , Mark SweetinghamAffiliated withDepartment of Agriculture and Food Western Australia
  • , Huaan YangAffiliated withDepartment of Agriculture and Food Western Australia Email author 

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Abstract

Seeds and plants of wild type Lupinus albus are bitter and contain high level of alkaloids. During domestication, at least three genes conferring low-alkaloid content were identified and incorporated into commercial varieties. Australian lupin breeders exclusively utilize one of these sweetness genes, “pauper”, in all varieties to prevent possible bitterness contamination via out-crossing. A cross was made between a sweet variety Kiev Mutant (containing pauper gene) and a bitter type landrace P27174, and the population was advanced into F8 recombinant inbred lines (RILs). Twenty-four plants representing sweetness and bitterness were subjected to DNA fingerprinting by the microsatellite-anchored fragment length polymorphism (MFLP) technique. A dominant polymorphism was discovered in an MFLP fingerprint. The MFLP marker was converted into a co-dominant, sequence-specific, simple PCR-based marker. Linkage analysis by the software program MapManager with marker score data and alkaloid phenotyping data from a segregating population containing 190 F8 RILs indicated that the marker is linked to the pauper gene at the genetic distance of 1.4 centiMorgans (cM). This marker, which is designated as “PauperM1”, is capable of distinguishing the pauper gene from the other two low-alkaloid genes exiguus and nutricius. Validation on germplasm from the Australian lupin breeding program showed that the banding pattern of the marker PauperM1 is consistent with the alkaloid genotyping on a wide range of domesticated varieties and breeding lines. The PauperM1 marker is now being implemented for marker assisted selection in the Australian albus lupin breeding program.

Keywords

Marker assisted selection (MAS) Molecular marker MFLP Lupinus albus