Strategies to obtain stable transgenic plants from non-embryogenic lines: complementation of the nn1 mutation of the NORK gene in Medicago sativa MN1008.
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.
Institute of Genetics, Biological Research Center of the Hungarian Academy of Sciences, Temesvari korut 62, 6726, Szeged, Hungary.
Ariana
A
Perhald
Gabriella
G
Endre
Zoltan
Z
Kevei
Gyorgy B
GB
Kiss
Attila
A
Kereszt
D000483:Alleles; D020090:Chromosome Segregation / Q000235:genetics; D032461:Chromosomes, Plant / Q000235:genetics; D018744:DNA, Plant / Q000378:metabolism; D047108:Embryonic Development; D017343:Genes, Plant / Q000235:genetics*; D005816:Genetic Complementation Test / Q000379:methods*; D006720:Homozygote; D000455:Medicago sativa / Q000235:genetics*; D009154:Mutation / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D030821:Plants, Genetically Modified; D019076:Transgenes
D018744:DNA, Plant