Strategies to obtain stable transgenic plants from non-embryogenic lines: complementation of the nn1 mutation of the NORK gene in Medicago sativa MN1008

Plant Cell Rep. 2006 Aug;25(8):799-806. doi: 10.1007/s00299-006-0117-6. Epub 2006 Mar 14.

Abstract

Strategies to introduce genes into non-embryogenic plants for complementation of a mutation are described and tested on tetraploid alfalfa (Medicago sativa). Genes conditioning embryogenic potential, a mutant phenotype, and a gene to complement the mutation can be combined using several different crossing and selection steps. In the successful strategy used here, the M. sativa genotype MnNC-1008(NN) carrying the recessive non-nodulating mutant allele nn ( 1 ) was crossed with the highly embryogenic alfalfa line Regen S and embryogenic hybrid individuals were identified from the F1 progeny. After transformation of these hybrids with the wild-type gene (NORK), an F2 generation segregating for the mutation and transgene were produced. Plants homozygous for the mutant allele and carrying the wild-type NORK transgene could form root nodules after inoculation with Sinorhizobium meliloti demonstrating successful complementation of the nn ( 1 ) mutation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Chromosome Segregation / genetics
  • Chromosomes, Plant / genetics
  • DNA, Plant / metabolism
  • Embryonic Development*
  • Genes, Plant / genetics*
  • Genetic Complementation Test / methods*
  • Homozygote
  • Medicago sativa / genetics*
  • Mutation / genetics*
  • Nucleic Acid Hybridization
  • Plants, Genetically Modified
  • Transgenes

Substances

  • DNA, Plant