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SCAR markerer of sorghum head smut resistance germ No. 3 physiological strain

A technology for anti-head smut and physiological races, applied in the direction of microbial measurement/testing, DNA/RNA fragments, recombinant DNA technology, etc., to shorten the breeding cycle, speed up the breeding process, and overcome the huge workload of auxiliary selection Effect

Inactive Publication Date: 2010-09-08
SHENYANG NORMAL UNIV
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  • Description
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  • SCAR markerer of sorghum head smut resistance germ No. 3 physiological strain
  • SCAR markerer of sorghum head smut resistance germ No. 3 physiological strain
  • SCAR markerer of sorghum head smut resistance germ No. 3 physiological strain

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Experimental program
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Embodiment 1

[0068] Embodiment 1: the acquisition of the RAPD polymorphic marker linked with the anti-head smut gene

[0069] The present invention uses 400 pairs of (S1-S400) RAPD primers to parent and its F 2 Generational populations were analyzed for markers. Among them, 355 pairs of primers amplified products, 45 pairs did not amplify products, and the amplification rate was 88.7%. Among them, primer S18 is in anti-parent, sense-parent and its F 2 A stable differential band was amplified from the generation population. According to Maker, the size of the fragment was about 850bp. 2 Two stable differential bands were amplified from the generation population, and according to Maker, the fragment sizes were about 1500bp and 700bp respectively (see figure 1 ), the co-segregation analysis of these three differential bands was carried out, and a total of 144 strains of F 2 Among them, there were 96 head smut-resistant strains and 48 head smut-sensitive strains. The analysis results showe...

Embodiment 2

[0070] Implementation Example 2: Recovery and sequencing of the RAPD marker linked to the head smut gene

[0071] S18 to be linked to head smut resistance gene 799、S336-1 1419 and S336-2 716 The three polymorphic fragments were retrieved and sequenced, and the sequencing results showed that the sizes of the three differential fragments were 799, 1419 and 716, so these three markers were named S18 799 、S336-1 1419 and S336-2 716 The sequencing results are as follows.

[0072] A. Polymorphic difference fragment S18 799 The sequencing result of:

[0073] 1 TCCACAGCAG TTGTACATCA GGTTTGCCTT CGTCTCCAAT CTCTTAGCAT

[0074] 51 TTTTCTCCTG GTTAATTTGA CTTCTCATTC GAGATGGGGA TAACTTTTGT

[0075] 101 TACTGTATGG TGCTATAATA ATTTCATCAT TTCTGAGCTT GTTGGTTTCT

[0076] 151 TTGTGGAACT ACTGAAATTG TGTTCACTAA TTCACTTGAT AGATTATGCT

[0077] 201 CTTAATAGAA ATTCAGCATA GTGCAGAGTG TCATTTCACT ATATTTTGTT

[0078] 251 ATTTTTAAGT AATATATGCT GGAGAGGGTT GCATCGCTTT TCCAAGCTGA

[0079] 301 GACTTAGGTT T...

Embodiment 3

[0138] Example 3: Conversion of RAPD markers to SCAR markers

[0139] According to the specific fragment S18 799 、S336-1 1419 and S336-2 716 Based on the principles of both-end sequence and primer design (avoid hairpin structure, appropriate G+C content), three pairs of specific primers were designed at both ends of the sequence (see Table 1 below for the primer sequence), and these three pairs of primers were used to detect 7050B (anti- parent), Tx622B (sense parent) and the F 2 The results showed that the SCAR marker and the RAPD marker of the head smut resistance gene had the same linkage in the resistant pool, sensitive pool and individual plants, and the PCR product was only a band of 799bp, 1419bp and 716bp, which were easy to distinguish. In this way, the RAPD marker was successfully transformed into a SCAR marker with good stability and repeatability (see Figure 4 , 5 , 6).

[0140] Table 1 SCAR primer sequence

[0141] Table 1 primer sequence of SCAR

[0142]...

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Abstract

The invention discloses an SCAR markerer of a sorghum head smut resistance germ No. 3 physiological strain, which takes 7050B which is immune to head smut, Tx622B which is high sensitive to the head smut and F2 hybridized by Tx622B / 7050B as test pieces and adopts a BSA method to make a molecular marker on a sorghum head smut resistance gene by applying an RAPD (Random Amplified Polymorphic DNA) screening technology. The invention establishes and optimizes a sorghum RAPD reaction system to obtain three RAPD markers of S18799, S336-11419 and S336-2716 which are tightly interlocked with the sorghum head smut resistance gene, also converts the S18799, the S336-11419 and the S336-2716 to stable SCAR markerers and uses the markers to carry out rapid PCR (Polymerase Chain Reaction) detection on the F2 generation and a part of sorghum resistant varieties and finds out a molecular marker specific band from the resistant varieties, thereby establishing an SCAR markerer rapid PCR detection method of the sorghum head smut resistance germ No. 3 physiological strain, supplying a basis for molecular marker assisted breeding and providing the technical support for practicing the molecular marker assisted breeding and shortening the breeding period.

Description

technical field [0001] The invention relates to a SCAR marker of sorghum anti-head smut physiological race No. 3, in particular a sorghum anti-head smut RAPD marker and a SCAR marker transformed from the marker, belonging to the field of biotechnology. Background technique [0002] Sorghum (Sorghum bicolor (L.) Moench) is one of the most important cereal crops in the world. It is mainly distributed in tropical arid and semi-arid regions, and is also planted in temperate and cold regions. From a global perspective, it is second only to wheat, rice, corn, and barley, ranking fifth. [0003] Sorghum head smut (Sphacelotheca reiliana (Kühn) Clinton) is a major sorghum disease worldwide. Sorghum head smut was first discovered in Egypt in 1868, and then spread to almost every sorghum producing area in the world. Head smut also occurs in all sorghum producing areas in China, and the damage is most serious in Northeast and North China, and it is one of the main diseases affecting ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 李玥莹邹剑秋郝林李雪梅马莲菊马纯艳陆丹彭霞陶思源陈琳
Owner SHENYANG NORMAL UNIV
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