Corn southern rust resistance gene and application thereof
A southern rust and maize technology, applied to southern rust resistance gene of maize and its application field, can solve the problem of single resistance source and the like
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Embodiment 1
[0061] Embodiment 1, discovery of resistance gene
[0062] Using corn inbred lines Jing 2416 and Jing 2416K as basic materials, configure F 1 hybrids, the F 1 The F of Jing 2416×Jing 2416K was obtained by inbred generation 2 Isolate populations for resistance gene mapping. All experimental materials were planted in Sanya, Hainan, where maize rust is high in the south. Sowing in mid-October, F 2 Group identification materials are randomly arranged and planted in a single row. The row length of each material was 5m, the row spacing was 0.55m, and 20-25 seedlings were reserved in each row. The maize inbred lines Jing 2416K and Jing 2416 were set as a group of high resistance and high sensitivity control materials in every 50 rows. Field management is carried out according to general field production.
[0063] Resistance identification was carried out in Sanya, Hainan by natural occurrence. The survey was carried out from the corn milk stage to the wax stage (about 15 days ...
Embodiment 2
[0081] Example 2, Application of Resistance Genes in Maize Genetic Improvement
[0082] 1. Preparation of transgenic corn
[0083] 1. Construction of recombinant vector
[0084] 1) The pYBA-p1132 vector was digested with restriction endonuclease EcoRI to obtain a linearized pYBA-p1132 vector, and the linearized vector was recovered by agarose gel.
[0085] The pYBA-p1132 vector is a vector obtained by inserting an expression cassette of a selection marker gene (herbicide resistance gene Bar gene) at position 1369 of the pYBA vector. The nucleotide sequence of the expression cassette of the screening marker gene (herbicide-resistant gene Bar gene) is shown in Sequence 5, which sequentially includes the 35S promoter, the screening marker gene Bar and the NOS terminator.
[0086] 2) Using the cDNA of Jing 2416K as a template, the primers pYBA-65F / R and pYBA-67F / R were used for PCR amplification respectively, and the full-length fragments of the coding region of the Zm00001d0232...
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