Localization of Ds-transposon containing T-DNA inserts in the diploid transgenic potato: Linkage to the R1 resistance gene against Phytophthora infestans (Mont) de Bary

被引:12
作者
ElKharbotly, A
Jacobs, JME
Hekkert, BTL
Jacobsen, E
Ramanna, MS
Stiekema, WJ
Pereira, A
机构
[1] AGR UNIV WAGENINGEN, DEPT PLANT BREEDING, GRAD SCH EXPT PLANT SCI, 6700 AJ WAGENINGEN, NETHERLANDS
[2] CTR PLANT BREEDING & REPROD RES CPRO DLO, DEPT MOLEC BIOL, 6700 AA WAGENINGEN, NETHERLANDS
关键词
Solanum tuberosum; Phytophthora infestans; Ds element; transposon tagging; R genes; euchromatin;
D O I
10.1139/g96-034
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The Dissociation transposable element (Ds) of maize containing NPTII was introduced into the diploid potato (Solanum tuberosum) clone J91-6400-A16 through Agrobacterium tumefaciens mediated transformation. Genomic DNA sequences flanking the T-DNAs from 312 transformants were obtained with inverse polymerase chain reaction or plasmid rescue techniques and used as probes for RFLP linkage analysis. The RFLP map location of 60 T-DNAs carrying Ds-NPTII was determined. The T-DNA distribution per chromosome and the relative distance between them appeared to be random. All 12 chromosomes have been covered with Ds-containing T-DNAs, potentially enabling tagging of any gene in the potato genome. The T-DNA insertions of two transformants, BET92-Ds-A16-259 and BET92-Ds-A16-416, were linked in repulsion to the position of the resistance gene R1 against Phytophthora infestans. After crossing BET92-Ds-A16-416 with a susceptible parent, 4 desired recombinants (Ds carrying T-DNA linked in coupling phase with the R1 gene) were discovered. These will be used for tagging the R1 gene. The efficiency of the pathway from the introduction to localization of T-DNAs is discussed.
引用
收藏
页码:249 / 257
页数:9
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