Corn southern rust resistance gene and application thereof

Abstract

The invention discloses a corn southern rust (Puccinia polysora Underw) resistance gene and application thereof. F2 segregation population of inbred line Jing 2416 and Jing 2416K is used for genetic analysis, and the result shows that the resistance gene contained in the Jing 2416K is a single dominant gene and is named as RppM. The application comprises the following steps of positioning RppM in an interval of a short arm 110-kb of a corn chromosome No. 10 through BSA re-sequencing analysis and genetic linkage analysis, performing function annotation analysis, sequence comparison analysis and expression quantity analysis on candidate genes in the interval, determining Zm00001d023265 and Zm00001d023267 as the candidate genes of the RppM, and performing molecular cloning on the two genes to obtain coding region sequences of the two genes. Transgenic experiment verification proves that the two genes both have the function of improving the resistance of the corn to the southern rust disease, and new resistant materials and gene resources are provided for corn resistance breeding.

Description

technical field

[0001] The invention relates to the technical field of plant genetic engineering, in particular to a maize southern rust resistance gene and its application. Background technique

[0002] Corn is the grain and economic crop with the largest planting area and the highest total output in my country, and occupies an important position in the national economy. Maize disease is an important factor affecting yield. With global climate change, changes in farming methods and the promotion of new varieties, the occurrence of corn diseases in my country has also changed. In the past, some secondary diseases (such as rust, tumor smut, etc.) pose a great threat. Southern corn rust is a fungal disease caused by Puccinia polysora Underw., which is mainly distributed in tropical and subtropical regions and is a very harmful airborne disease. In recent years, the disease has a tendency to spread to the north of our country. Once it breaks out and becomes popular, it will g...

Examples

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...

technical field

[0001] The invention relates to the technical field of plant genetic engineering, in particular to a maize southern rust resistance gene and its application. Background technique

[0002] Corn is the grain and economic crop with the largest planting area and the highest total output in my country, and occupies an important position in the national economy. Maize disease is an important factor affecting yield. With global climate change, changes in farming methods and the promotion of new varieties, the occurrence of corn diseases in my country has also changed. In the past, some secondary diseases (such as rust, tumor smut, etc.) pose a great threat. Southern corn rust is a fungal disease caused by Puccinia polysora Underw., which is mainly distributed in tropical and subtropical regions and is a very harmful airborne disease. In recent years, the disease has a tendency to spread to the north of our country. Once it breaks out and becomes popular, it will g...

Examples

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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