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Insect resistance. Hessian fly

Gene Hdicoccum

Protocol contributed by X. Lui, M. Chen and
G. Brown-Guedira (Gina_Brown-Guedira@ncsu.edu)

Hessian fly resistance gene "Hdicoccum" (or Hdic) was transferred to wheat germplasm KS99AGRC42 from PI 94641, an accession of cultivated emmer wheat (T. turgidum ssp. dicoccum). The resistance of PI 94641 to Hessian fly is controlled by a single dominant gene located on chromosome 1AS. Thirteen genes for resistance to Hessian fly have been transferred to common wheat from durum wheat (T. turgidum ssp. durum), a species closely related to emmer wheat. Two of these genes, H5 and H11 are also located on the short arm of wheat chromosome 1A. The Hessian fly resistance gene in PI 94641 is effective at high temperature. When evaluated for reaction to Biotype L of the Hessian fly at high temperatures, lines having H5 and H11 were susceptible, indicating that the gene in KS99WGRC42 is different from H5 and H11. It is not known if the resistance gene in KS99WGRC42 is allelic to either H5 or H11.

Microsatellite markers Xgwm136 and Xbarc263

Microsatellite markers flanking Hessian fly resistance gene "Hdicoccum" have been identified. Xgwm136 is a recessive marker 7.4 cM from the gene and Xbarc263 is a co-dominant marker 4.9 cM from the gene (1).

Locus Xgwm136-1A

PCR primers:
    GWM136-L   5'- GAC AGC ACC TTG CCC TTT G -3'
    GWM136-R   5'- CAT CGG CAA CAT GCT CAT C -3'   
PCR conditions:
  • Denaturing step: 3 min at 94°C
  • 45 cycles of: [1 min at 94°C, 1 min at 60°C, 2 min at 72°C ]
  • Extension step: 10 min at 72°C

Locus Xbarc263-1A

PCR primers
    BARC263-L   5'- GGA AGC GCG TCA GCA CTA GGC AAC -3'   
    BARC263-R   5'- GGC TTC TAG GTG CTG CGG CTT TTG TC -3'   
PCR conditions:
  • Denaturing step: 3 min at 95°C
  • 35 cycles of: [40 sec at 94°C, 40 sec at 55°C, 1 min at 72°C ]
  • Extension step: 10 min at 72°C

Expected products

Xgwm136-1A. Polymorphic DNA fragment amplified using primer pair GWM136.

Karl92 (K) and Jagger (J), and the resistance gene donor parent, the emmer wheat PI 94641 (PI).
R: Hessian Fly (HF) resistant line, S: Hessian Fly susceptible line, H: segregating for HF resistance.
A 320-bp band (blue arrow) present in the S parents (K, J) was absent in the R parent (T). This marker behaves in a recessive fashion and cannot be used to detect Hdicoccum in BCF1 plants.


Xbarc263-1A. Polymorphic DNA fragments amplified using primer pair BARC263.

Karl92 (K) and Jagger (J), and the resistance gene donor parent the emmer wheat (Triticum turgidum subsp. dicoccum) PI 94641 (PI).
R: Hessian Fly (HF) resistant line, S: HF susceptible line, H: segregating for HF resistance.
The 210-bp band (blue arrow) was present in the S parents (K, J) and the 220-bp band (red arrow) was present in the R parent (PI).


Conditions presented here should be considered only as a starting point of the PCR optimization for individual laboratories.

References

1. Genetic characterization and molecular mapping of a Hessian fly-resistance gene transferred from T. turgidum ssp. dicoccum to common wheat. Liu XM, Brown-Guedira GL, Hatchett JH, Owuoche, Chen MS. In: Theoretical and Applied Genetics, 2005, 111:1308-1315. DOI: 10.1007/s00122-005-0059-3

 

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