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CN-122012780-A - Method for screening waxy wheat gene Wx-B1B by using powdery mildew resistance gene MlIW30

CN122012780ACN 122012780 ACN122012780 ACN 122012780ACN-122012780-A

Abstract

The invention discloses a method for screening waxy wheat gene Wx-B1B by using powdery mildew resistance gene MlIW, belonging to the technical field of biology. The invention solves the technical problem of how to screen Wx-B1B loci. The method for screening waxy wheat gene Wx-B1B comprises the steps of detecting powdery mildew resistance of wheat to be detected, and detecting whether a Wx-B1 site carries Wx-B1B by using powdery mildew disease resistance phenotype, wherein the Wx-B1 site of the wheat to be detected resistant to powdery mildew carries Wx-B1B, the Wx-B1B is a DNA fragment shown as SEQ ID No.7, and the Wx-B1a is a DNA fragment shown as SEQ ID No. 8. The method can be used for breeding high-efficiency polymeric materials with disease-resistant waxy sites in the traditional breeding process.

Inventors

  • XIE CHAOJIE
  • SUN JUNNA
  • PAN WEI
  • MA JUN
  • HU ZHAORONG

Assignees

  • 中国农业大学

Dates

Publication Date
20260512
Application Date
20260211

Claims (10)

  1. 1. A method for detecting a wheat Wx-B1 locus, wherein the Wx-B1 locus is a DNA fragment corresponding to SEQ ID No.7 or SEQ ID No.8 in a wheat genome and is Wx-B1B or Wx-B1a, the method comprises detecting powdery mildew resistance of wheat to be detected, and detecting whether the Wx-B1 locus carries Wx-B1B by using a powdery mildew disease resistance phenotype, wherein the Wx-B1 locus of the wheat to be detected resistant to powdery mildew carries Wx-B1B; The Wx-B1B is a DNA fragment shown in SEQ ID No. 7; The Wx-B1a is a DNA fragment shown in SEQ ID No. 8.
  2. 2. The method of claim 1, wherein the detection of powdery mildew resistance of the wheat under test is accomplished by inoculating powdery mildew to the wheat under test to determine powdery mildew resistance.
  3. 3. The method of claim 1, wherein the detection of powdery mildew of wheat to be detected comprises PCR amplification of genomic DNA of the wheat to be detected by using a primer pair consisting of IW30InDel-F and IW30InDel-R, wherein the IW30InDel-F and the IW30InDel-R are single-stranded DNA shown as SEQ ID No.1 and SEQ ID No.2 respectively; The wheat to be tested of which the PCR product contains the DNA fragment 1431 bp is or is candidate to be powdery mildew resistant wheat, and the wheat to be tested of which the PCR product does not contain the DNA fragment 1431 bp is or is candidate to be non powdery mildew resistant wheat.
  4. 4. Use of the method of any one of claims 1-3 for cultivating waxy wheat, or for cultivating powdery mildew resistant waxy wheat.
  5. 5. Use of a substance detecting a wheat MlIW locus in detecting a wheat Wx-B1 locus, wherein the Wx-B1 locus is a DNA fragment corresponding to SEQ ID No.7 or SEQ ID No.8 in the wheat genome, which is Wx-B1B or Wx-B1a; The Wx-B1B is a DNA fragment shown in SEQ ID No. 7; The Wx-B1a is a DNA fragment shown in SEQ ID No. 8.
  6. 6. The method according to claim 5, wherein the substance for detecting MlIW locus is a primer pair consisting of single-stranded DNA shown in SEQ ID No.1 and SEQ ID No. 2.
  7. 7. The application of the substance for detecting the MlIW locus of wheat in cultivating waxy wheat or in cultivating powdery mildew resistant waxy wheat.
  8. 8. The method according to claim 7, wherein the substance for detecting the MlIW locus of wheat is a primer pair consisting of single-stranded DNA shown in SEQ ID No.1 and SEQ ID No. 2.
  9. 9. The application of the substance for detecting the wheat MlIW locus and the substance for detecting the wheat Wx-B1 locus in the cultivation of waxy wheat or the cultivation of powdery mildew resistant waxy wheat.
  10. 10. The application of claim 9, wherein the substance for detecting the MlIW locus of wheat is a primer pair consisting of single-stranded DNA shown in SEQ ID No.1 and SEQ ID No. 2; The substances for detecting the wheat Wx-B1 locus are primers shown as SEQ ID No.3, SEQ ID No.4, SEQ ID No.5 and SEQ ID No. 6.

Description

Method for screening waxy wheat gene Wx-B1B by using powdery mildew resistance gene MlIW30 Technical Field The invention belongs to the technical field of biology, and particularly relates to a method for screening waxy wheat gene Wx-B1B by using powdery mildew resistance gene MlIW. Background Wheat is used as an important grain crop, the production of the wheat is limited by climate change and threatened by pathogen small body renewal and toxicity enhancement, the wheat powdery mildew is an air-borne fungus disease caused by the powdery mildew of the brucella gramineae, the epidemic range is wide, the variation is rapid, and the sustainable production [1] of a main wheat production area of the world is threatened. The resistance of wheat to powdery mildew is determined by disease-resistant genes, which disease-resistant genes are carried is the key for determining whether the wheat is resistant to disease or not, the breeding of disease-resistant varieties by using the disease-resistant genes is the most economical and effective measure for preventing and treating the wheat powdery mildew, the traditional breeding method has long selection period and low efficiency, and the molecular marker is used for assisting in breeding new wheat germplasm polymerized by the disease-resistant genes and other high-quality genes, so that the breeding period can be shortened, and the accuracy of gene selection can be improved. Powdery mildew resistance gene MlIW from wild two-grain wheat (Triticum dicoccoides, 2n=4x=28, aabb) showed full-growth resistance to physiological race E09 prevalent in beijing area, was located [2] on chromosome 4AL, and was accurately identified by its tightly linked molecular marker IW30InDel (CN 120591444 a). The wild two-grain wheat can be fully recombined into the genome of the common wheat as ancestor of the common wheat, enriches the genetic background of the wheat and has great breeding value. Starch is an important component of wheat grain, including both amylopectin and amylose, accounting for 20% -30% and 70% -80% of the total starch content, respectively, and is referred to as full waxy wheat when the amylose content in wheat grain is < 1%. Japanese scholars in 1993 have found that wheat amylose synthesis is controlled primarily by granule-bound STARCH SYNTHASE I (GBSSI), also known as the wall protein [3]. The content of the wall protein is positively correlated with the Waxy and non-Waxy properties of wheat, and the gene encoding the wall protein is named as Wx gene. The wheat of the common hexaploid contains 3 w xy protein subunits, the Waxy property of the wheat is controlled and synthesized by Wx-A1, wx-B1 and Wx-D1 genes positioned on 7AS, 4AL and 7DS, and the corresponding allelic deletions are called Wx-A1B, wx-B1B and Wx-D1B [4]. Chen Dongsheng and other studies show that the Wx gene in common wheat endosperm has a dose effect on amylose content, namely the wheat amylose content is reduced most when the Wx-B1 subunit is absent, the influence on the amylose content is minimum when the Wx-A1 and the Wx-D1 are simultaneously absent when two Wx protein subunits are absent, and the wheat endosperm almost does not contain amylose when 3 Wx protein subunits are completely absent, which is also called full waxy wheat [5]. In 1999, murai published DNA sequences of 3 Wx genes in wheat, wx-A1 is 2791 bp, wx-B1 is 2794 bp, wx-D1 is 2862 bp, and the similarity of the encoded proteins is as high as 96.5% -97.4% [6]. It has been found that Wx-A1 and Wx-D1 site variations of the Wx gene are not permanent and comprise base insertions and deletions in many cases, and that the Wx-B1 site is a deletion of the complete coding sequence. The proportion of the wheat amylose and the amylopectin affects the viscosity, gelatinization property and the yield of starch, waxy wheat is taken as a special wheat, the amylopectin content is high, so that the waxy wheat has higher starch viscosity, good ductility, obvious advantages in the aspects of making noodles, special cakes and the like, the aging delay effect of the made bread is good, the dough viscosity is rapidly increased when the temperature is above 60 ℃, and the waxy wheat can be taken as a material [7], liang Rongji of cooked foods and the like, and researches show that the low amylose variety is not easy to sprout, the yield loss before harvest is reduced, and the flour quality reduction [8] caused by the increase of amylase activity in the processing is avoided. In addition, the polymer has important value in paper making and textile industry, and can be made into degradable agricultural films, so that white pollution in agriculture in recent years is reduced. The detection of waxy wheat mainly comprises the modes of iodine staining method, SDS-PAGE, far infrared spectrum, antibody, 2D-PAGE, molecular marking and the like, wherein the iodine staining method only can identify full waxy wheat, waxy wheat is stained in reddish brown, waxy wheat is