Breeding method utilizing wild rice resource to create japonica-type novel paddy rice species

A technology for wild rice and new germplasm, applied to other methods of inserting foreign genetic materials, using microinjection methods, botanical equipment and methods, etc., can solve the problems of sterility of hybrid offspring, narrow genetic basis, and lack of flowering. To achieve the effect of shortening the breeding period, broad application prospects, and rapid stability of offspring

Active Publication Date: 2014-05-28
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AI-Extracted Technical Summary

Problems solved by technology

At present, the genetic basis of japonica rice cultivars is narrow and their stress resistance is poor. The hybridization of wild rice and japonica rice has p...
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The invention belongs to the fields of paddy rice improvement and biological technology application, and specifically relates to a breeding method utilizing wild rice resource to create japonica-type novel paddy rice species. The method comprises the following steps: S1, extracting DNA genome from wild rice; S2, introducing DNA into paddy rice; S3, screening the plants with varied characters; S4, carrying out molecular verification on the screened variant plants. The invention provides a practicable and feasible way for creating novel paddy rice species. The method introduces a wild rice DNA into japonica-type rice so as to obtain an offspring material having wild rice characters of the japonica-type rice, and the introduction efficiency is as high as 6.8%. The method has the advantages of wide variation, wide selection population, shortened breeding period, suitability for breeding needs, and quick character stability, and the third generation of the introduced rice has stable varied characters. The created novel species can be used as a parent material for japonica rice breeding, or is subjected to a directed breeding to generate a novel species which can be applied to paddy rice production.

Application Domain

Microinjection basedAngiosperms/flowering plants

Technology Topic

Japonica riceParent material +7


  • Breeding method utilizing wild rice resource to create japonica-type novel paddy rice species
  • Breeding method utilizing wild rice resource to create japonica-type novel paddy rice species
  • Breeding method utilizing wild rice resource to create japonica-type novel paddy rice species


  • Experimental program(1)
  • Effect test(1)

Example Embodiment

[0026] The following describes the present invention in further detail with reference to the embodiments, but it is not a limitation of the present invention. Any equivalent replacement in the field made according to the disclosure of the present invention belongs to the protection scope of the present invention. Example 1
[0027] The experimental materials in this example are wild rice DNA, Ningjing 16 and Ningjing 23. The breeding method of using wild rice resources to create new germplasm of japonica rice in this embodiment includes the following steps:
[0028] The present invention uses wild rice resources to create a new breeding method of japonica rice germplasm, which includes the following steps:
[0029] S1. Extraction of Wild Rice Genomic DNA
[0030] (A) Cut 5g of leaves at the booting stage into a 50ml mortar;
[0031] (B) Add liquid nitrogen to the mortar of step (a), add 50ml each time, 3 to 4 times in total, quickly grind into fine powder, and pour the fine powder into a 50ml plastic centrifuge tube;
[0032] (C) Add 25ml of preheated extraction buffer and 25μl of β-mercaptoethanol to the centrifuge tube of step (b), mix well and water bath at 65℃ for 30min; extraction buffer 100mmol/L Tris-HCl pH8.0, 500mmol/L NaCl, 50mmol/L EDTA pH8.0, mass concentration is 1.5% SDS.
[0033] (D) Add 5ml of 5mol/L potassium acetate solution to the centrifuge tube of step (c), mix gently, and ice bath for 20min;
[0034] (E) Add an equal volume of a mixture of chloroform and isoamyl alcohol to the centrifuge tube of step (d), the volume ratio of chloroform to isoamyl alcohol is 24:1, and centrifuge at 4500×g at 4°C for 15 minutes;
[0035] (F) Aspirate the supernatant in step (e), add an equal volume of isopropanol pre-cooled at -20°C, and stand still for 3min-5min until white DNA coils appear;
[0036] (G) Pick up the DNA coil in step (f), wash with 70% ethanol, dry, and dissolve in 0.1ml 1×SSC, store at 4°C;
[0037] S2. DNA import
[0038] In the late booting stage, the pollen develops to the middle and late stages of mononucleus, and the external morphology is that the top of the panicle is 2cm from the flag leaf occiput. The DNA is introduced into the neck of the panicle with a micro-injector, and it penetrates into the neck section of the panicle at a 45 degree angle from top to bottom. Place, inject DNA solution, the DNA concentration is 400μg/ml, and the buffer solution is 2mg/ml CaCl added 2 0.1×SSC buffer, the injection time is from 9 am to 10 am, each plant is introduced 10μl, and the introduction is repeated 3 times; the introduced ears are harvested after the introduced plants mature, and the mutants are planted next year;
[0039] S3. Screening plants with variant traits
[0040] The harvested introduced panicles were cultivated for soilless seedlings according to panicles and control panicles. When the seedlings grew to about 10cm, they were planted into the field by panicles. The recipient materials were used as controls to select plants with variant traits, and continue with the variants. Plant until the traits of the offspring are stable.


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