Maize southern rust resistance gene and its application
A technology for southern rust and corn, which is applied in the field of corn southern rust resistance gene and its application, and can solve the problem of single source of resistance
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Embodiment 1
[0061] Example 1. Discovery of resistance genes
[0062] Based on corn inbred lines Jing 2416 and Jing 2416K, configuration F 1 Hybrid, the F 1 Obtained F of Jing 2416×Jing 2416K on behalf of self-introduction 2 Populations were segregated for resistance gene mapping. All experimental materials were planted in Sanya, Hainan, a high incidence area of southern corn rust. Sow mid-October, F 2 The population identification materials were randomly arranged and planted in a single row. Each material was sown with a row length of 5m and a row spacing of 0.55m, with 20 to 25 seedlings in each row. Maize inbred lines Jing 2416K and Jing 2416 were set up in 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, relying on natural disease. Surveys were conducted from the milk to wax stage of corn (approximately 1...
Embodiment 2
[0081] Example 2. Application of resistance gene 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 the linearized pYBA-p1132 vector, and the linearized vector was recovered from agarose gel.
[0085] The pYBA-p1132 vector is a vector obtained by inserting an expression cassette of a selectable marker gene (herbicide resistance gene Bar gene) at position 1369 of the pYBA vector. The nucleotide sequence of the expression cassette of the selectable marker gene (herbicide resistance gene Bar gene) is shown in SEQ ID NO: 5, which sequentially includes 35S promoter, selectable marker gene Bar and NOS terminator.
[0086] 2) Using the cDNA of Jing 2416K as a template, PCR amplification was carried out with primers pYBA-65F / R and pYBA-67F / R, respectively, and the full-length fragment of the coding region of Zm00001d023265 g...
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