Breeding method for directed improvement of tobacco black shank resistance by molecular marker-assisted selection

A molecular marker-assisted, tobacco black shank technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the loss of black shank resistance, the inability to maintain safflower, and field characteristics. The problem of heavy workload, etc., can achieve the effect of increasing the resistance to black shank disease, improving the background recovery rate, and growing neatly in the field.

Active Publication Date: 2017-03-15
YUNNAN ACAD OF TOBACCO AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For more than 20 years, Chinese tobacco breeders have tried to use conventional breeding techniques to improve the resistance of Honghua Dajinyuan to black shank, but they have not been successful. The selected offspring materials cannot maintain the characteristics of Honghuadajinyuan varieties. characteristics and the loss of blackleg resistance; and

Method used

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  • Breeding method for directed improvement of tobacco black shank resistance by molecular marker-assisted selection
  • Breeding method for directed improvement of tobacco black shank resistance by molecular marker-assisted selection
  • Breeding method for directed improvement of tobacco black shank resistance by molecular marker-assisted selection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1: Acquisition of foreground and background molecular markers

[0055] The 23626 SSR markers developed with the whole genome data of Honghua Dajinyuan were used to test the recurrent parent Honghua Dajinyuan, the donor parent RBST (Chinese invention patent ZL201410819251.2) and its f 1 The DNA extracted from the leaves was screened for molecular markers in the laboratory of No. 14 Nanxiang Road, Yuxi City, Yunnan Academy of Tobacco Agricultural Sciences. 1 601 co-dominant molecular markers were used in the map software Joinmap 4.0 to compare 601 markers in 213 BC strains obtained in Datian, Yunnan Academy of Tobacco Agricultural Sciences Research and Experimental Base 1 f 1 The mapping population was subjected to genotype analysis of molecular markers and the construction of a genetic linkage map. Finally, a high-density genetic map covering the entire tobacco genome containing 581 SSR markers was constructed. Combined with 3 batches of 8030 BC plants in the g...

Embodiment 2

[0056] Example 2: BC 1 f 2 marker assisted selection

[0057] BC in 2012 1 f 1 BC obtained by selfing of resistant single plant 1 f 2 , BC 1 f 2 Seedlings were sown in 3 trays (128 plants / tray), transplanted to small flower pots after seedlings were grown, and then identified for resistance to blackleg disease after returning seedlings. A single plant without disease was transplanted to the field, and a total of 90 plants were transplanted. Leaf DNA of a single plant without disease; first use the four foreground markers Scf_30K, TM62, ST141 and InDel0617 that are closely linked to both sides of the blackleg resistance gene to help prospect selection, select a single plant containing the blackleg resistance gene, and then use the 24 linkage groups on the genome-wide genetic map of the improved original variety, and 5 background markers were selected for each linkage group, a total of 120 background markers evenly distributed in each linkage group assisted background sel...

Embodiment 3

[0058] Example 3: BC 2 f 1 marker assisted selection

[0059] BC 2 f 1Seedlings were sown in 5 trays (168 plants / tray), transplanted to small flowerpots after seedlings were grown, and identified for resistance to black shank after returning seedlings. A single plant without disease was transplanted to the field, and a total of 306 plants were transplanted. Leaf DNA of a non-infested individual plant; first use 4 foreground markers Scf_30K, TM62, ST141 and InDel0617 closely linked to both sides of the black shank resistance gene to assist foreground selection, and select a single plant containing the black shank resistance gene; based on construction 24 linkage groups on the whole genome genetic map of the original variety, and 5 background markers were selected for each linkage group, a total of 120 background markers evenly distributed in each linkage group were used to assist in the preliminary selection of the background. 33 individual plants with high recovery rate; t...

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Abstract

The invention discloses a breeding method for directed improvement of tobacco black shank resistance by molecular marker-assisted selection. The breeding method includes the steps: acquiring a molecular marker; performing molecular marker-assisted selection for five times. Existing excellent characteristics of Honghua Dajinyuan can be completely preserved, black shank resistance is enhanced, field growth is regular, and genetic characters are stable. A progressive two-step method assists in background selection, so that the workload of background molecular marker-assisted selection can be effectively decreased. The background recovery ratio of single plants with backcross 3-generation BC3F1 subjected to molecular marker-assisted selection can reach 97.5% or more and is greatly increased, and the breeding process is shortened. The harmful or adverse effect of foreground linkage drag on improved varieties is decreased by selection, and the excellent characteristics of improved initial varieties are effectively and comprehensively preserved. Large-scale field planting for phenotype screening is omitted in the whole directed improvement process, field workload is low, most of work is less affected by environments, and results are more accurate.

Description

technical field [0001] The invention belongs to the technical field of tobacco disease-resistant breeding, and in particular relates to a breeding method for directional improvement of tobacco black shank resistance by molecular marker-assisted selection. Background technique [0002] The development of molecular biology technology, especially molecular marker technology, provides an effective means for variety improvement. Molecular marker-assisted selection breeding is to select target traits with the help of molecular markers closely linked to target genes during the breeding process. The major project of the China Tobacco Genome Project launched in 2010 has drawn a set of high-quality genome maps centered on the whole genome sequence maps of Nicotiana tomentosa, tobacco forest, and cultivated tobacco, tobacco molecular genetic linkage maps, and tobacco haplotype maps. Laid a solid foundation for molecular marker-assisted breeding; [0003] The flue-cured tobacco variet...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 肖炳光李永平童治军陈学军方敦煌焦芳婵王德勋徐兴阳范志勇端永明
Owner YUNNAN ACAD OF TOBACCO AGRI SCI
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