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Functional molecular marker of triticum aestivum L. anti-blumeria graminis relevant gene Pm41, and application of functional molecular marker

A powdery mildew technology with the same function, applied in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of insufficient practicability and accuracy, long distance, etc., and achieve important theoretical significance and economic value. , the effect of accelerating the pace of breeding

Active Publication Date: 2021-01-05
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the distance between these molecular markers and the Pm41 gene, their practicability and accuracy in molecular marker-assisted selection breeding are insufficient

Method used

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  • Functional molecular marker of triticum aestivum L. anti-blumeria graminis relevant gene Pm41, and application of functional molecular marker
  • Functional molecular marker of triticum aestivum L. anti-blumeria graminis relevant gene Pm41, and application of functional molecular marker
  • Functional molecular marker of triticum aestivum L. anti-blumeria graminis relevant gene Pm41, and application of functional molecular marker

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Embodiment 1, wheat powdery mildew resistance gene Pm41 (hereinafter referred to as Pm41 gene) high-density genetic linkage map construction

[0051] 1. Crossbreed with TZ-2 as the male parent and Langdon as the female parent to obtain the hybrid F 1 generation.

[0052] 2. After completing step 1, use the wheat seedling stage powdery mildew resistance identification method to detect TZ-2, Langdon or hybrid F 1 Powdery mildew resistance of generation; adopt Coomassie brilliant blue staining method (recorded in the following documents: Li Jianqiang, Liu Xili and Wang Hongmei (2002) Fluorescein sodium and Coomassie brilliant blue are applied to wheat powdery mildew bacteria staining effect comparison. Bacterial system, 21( 4):592-596) detection of TZ-2, Langdon or hybrid F 1 generations of powdery mildew resistance.

[0053] See the test results figure 1 (F 1 for hybrid F 1 Generation, A is the reaction of leaves inoculated with powdery mildew, B is the staining of ...

Embodiment 2

[0061] Embodiment 2, Pm41 gene physical map construction

[0062] 1. Construction of the BAC library of the powdery mildew-resistant parent TZ-2 (recorded in the following literature: Liang Yong. Construction of the physical map of the wild emmer wheat powdery mildew resistance gene MlIW170 and comparative analysis of the homologous regions of the 2DS of Goatwort rough [D]; China Agricultural University; 2015). The constructed BAC library of powdery mildew-resistant parent TZ-2 contained 326,784 clones in total, with an average insert length of 120 kb, about 3× genome coverage, and contained HindIII restriction sites. Positive clones from the BAC library were aliquoted in 851 384-well plates. All the BAC clones in each 384-well plate were mixed, and after mixing, the bacteria were shaken to extract the BAC plasmid, and a primary screening pool was established, with a total of 851 pools. After numbering the 851 mixed pools, place them in a 100-well box of 10×10. In each 100-w...

Embodiment 3

[0065] Embodiment 3, expression analysis of CNL gene

[0066] hpi (hour post inoculation) indicates the time after inoculation.

[0067] The primers for detecting CNL gene or cnl gene are 41R-315 shown in Table 1. The primers for detecting Actin gene are Actin shown in Table 1.

[0068]1. When TZ-2 or Langdon grows to 2 leaves, use powdery mildew to inoculate with powdery mildew (recorded in the following documents: liu ZY, Sun QX, Ni ZF, Yang TM(1999) Development of SCArmarkers linked to the Pm21gene conferring resistance to powdery mildew in common wheat. Plant Breed 118:215-219).

[0069] 2. After completing step 1, take leaves at 0hpi, 2hpi, 4hpi, 12hpi, 18hpi, 24hpi, 36hpi, 48hpi, 60hpi and 72hpi time points respectively, then extract RNA, reverse transcribe to obtain cDNA.

[0070] 3. Use the cDNA obtained in step 2 as a template for semi-quantitative detection.

[0071] For semi-quantitative test results, see Figure 4 Middle A (CNL TZ-2 Indicates the CNL gene in ...

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Abstract

The invention discloses a functional molecular marker of a triticum aestivum L. anti-blumeria graminis relevant gene Pm41, and application of the functional molecular marker. The genome DNA of triticum aestivum L. is taken as a template, and a DNA fragment obtained by amplification of a primer pair formed by a primer F and a primer R is a molecular marker Pm41-427. The primer F is a single-stranded DNA molecule disclosed by a sequence 8 of a sequence table, and the primer R is a single-stranded DNA molecule disclosed by a sequence 9 of the sequence table. When the molecular marker Pm41-427 provided by the invention is applied, a triticum aestivum L. material with blumeria graminis resistance can be quickly screened so as to quicken a breeding pace of a new variety of the triticum aestivumL.. The functional molecular marker has an important theoretical meaning and economic value.

Description

technical field [0001] The invention belongs to the field of plant molecular breeding, and in particular relates to a functional molecular marker of wheat powdery mildew resistance-related gene Pm41 and its application. Background technique [0002] Wheat (Triticum aestivum L.) is one of the three major staple foods in the world, and more than one-third of the world's population uses it as an important source of food. Powdery mildew is a fungal disease that seriously harms wheat production, and its causative bacterium is the gramineous powdery mildew (Blumeria graminis f.sp.tritici). Infection of powdery mildew in wheat will cause photosynthetic efficiency to decrease, which will affect the normal growth and development of plants. At the same time, it will also cause the rate of tillering and ear formation to decrease, and the thousand-grain weight will decrease, resulting in reduced yield. Production practice shows that compared with spraying chemical agents, cultivating l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 刘志勇李淼淼王振忠李贝贝董玲丽陆平吴秋红陈永兴
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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