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A complete set of primers for identification or auxiliary identification of Fusarium wilt of banana and its application

A technique for identifying and assisting identification of Fusarium wilt of banana, a complete set of primers for identification or auxiliary identification of Fusarium wilt of banana and its application field

Active Publication Date: 2017-01-18
ENVIRONMENT & PLANT PROTECTION INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A physiological specialization may cause disease on a particular plant species, but strains belonging to other specializations may be harmless or even beneficial to the same particular plant species

Method used

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  • A complete set of primers for identification or auxiliary identification of Fusarium wilt of banana and its application
  • A complete set of primers for identification or auxiliary identification of Fusarium wilt of banana and its application
  • A complete set of primers for identification or auxiliary identification of Fusarium wilt of banana and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0112] Example 1. Preparation of primers for identification or auxiliary identification of banana Fusarium wilt

[0113] A pair of PCR primers for identifying Cuban-specific Fusarium oxysporum in banana Fusarium wilt. The name is B. A 730bp DNA can be amplified from the Cuban-specific DNA of Fusarium oxysporum in banana Fusarium wilt. Fragment, a 699bp DNA fragment can be amplified from the Cuban-specific RNA of Fusarium oxysporum in Banana Fusarium wilt or cDNA transcribed from RNA as a template; the B is represented by SEQ ID No. in the sequence table. 3 and SEQ ID No. 4 shown in two single-stranded DNA composition.

[0114] The PCR primer pair for identifying the Cuban-specific race 4 of Fusarium oxysporum in banana Fusarium wilt, whose name is A, can be derived from the Fusarium oxysporum Cuban-specific race 4 in banana Fusarium wilt A DNA fragment with a size of 699 bp is amplified from DNA or RNA or cDNA transcribed using RNA as a template; said A is composed of two single-s...

Embodiment 2

[0117] Example 2. Specificity and sensitivity experiment of PCR primers for identification of Fusarium oxysporum Cuban-specific type of Fusarium oxysporum in Example 1 to B

[0118] 1. Preparation of template

[0119] According to the literature (Yang Laying, Huang Huaping, Tang Furun, et al. Acta Phytopathology. 2006, 36(3): 219-225.), the total DNA of the following strains was extracted and obtained:

[0120] 1: B2; 2: B2R1; 3: B2gfp; 4: BW4; 5: TR4; 6: STR4; 7: 3#①; 8: 3#②; 9: 5#①; 10: 5#②; 11: 6#①;12:6#②;13:7#①;14:7#②;15:9#①;16:9#②;17:34#①;18:34#②;19:36# ①; 20: 36#②; 21: 37#①; 22: 37#②; 23: QWXZ b-2; 24: NXXZ; 25: NBXZ a; 26: DTXZ-1 a-1; 27: DTXZ-2 Dimension a; 28: DTXZ-3 dimension; 29: LDXZ a; 30: LGXD-5 dimension; 31: LGXD-6 dimension; 32: ZJXD; 33: ZJB1 b1; 34: ZJB2 b2; 35: XNB2 a; 36: XNB5 a; 37: XNB6 a; 38: XSBN; 39: HH; 40: HZ; 41: ZJ; 42: NX-27; 43: PB3-15; 44: 11#②; 45: 40#①; 46: 40#②; 47: 42#①; 48: N2; 49: BXFZ-1 b-1; 50: BXFZ-2a-2; 51: FOC1; 52: Foc1 e2; 53: Race1; ...

Embodiment 3

[0140] Example 3. Specificity and sensitivity experiment of PCR primers for identification of Fusarium oxysporum race 4, race 4 in banana Fusarium wilt of example 1

[0141] 1. The specificity experiment of PCR primers for identification of Fusarium oxysporum race 4, race 4 in banana Fusarium wilt in Example 1

[0142] Using the total DNA of the strain and the cDNA of the strain as the templates for PCR amplification, respectively, the PCR primers of the identification of Fusarium oxysporum in the banana Fusarium wilt of Example 1 and the Cuba specific race 4 were specifically identified for A , The experiment is repeated three times, the specific steps of each repeated experiment are as follows:

[0143] 1. Use the total DNA of the strain as the template for PCR amplification

[0144] Using the PCR primers of Example 1 to identify the Fusarium oxysporum race 4 of Fusarium oxysporum in Banana Fusarium wilt, the total DNA of each strain in step 1 of Example 2 was amplified by PCR. The...

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Abstract

The invention discloses complete set primers for identification or assisted identification of fusarium oxysporum f.sp.cubense and application of the complete set primers. The complete set primers for identification or assisted identification of fusarium oxysporum f.sp.cubense provided by the invention comprise a complete set primer 1, a complete set primer 2 or a complete set primer 3, wherein the complete set primer 1 consists of a PCR primer pair named A, a PCR primer pair named B and a PCR primer pair named C; the A consists of two DNA single strands shown in SEQ ID No.1 and SEQ ID No.2 in a sequence table; the B consists of two DNA single strands shown in SEQ ID No.3 and SEQ ID No.4 in the sequence table; the C consists of two DNA single strands shown in SEQ ID No.5 and SEQ ID No.6 in the sequence table; the complete set primer 2 consists of A and C; and the complete set primer 3 consists of B and C.

Description

Technical field [0001] The invention relates to a set of primers and applications for identifying or assisting in identifying banana Fusarium wilt in the field of biotechnology. Background technique [0002] Fusarium oxysporum (Fusarium oxysporum) is a soil-borne pathogenic fungus with a wide range of hosts, which can cause wilt of more than 100 plants such as melons, Solanaceae, bananas, cotton, legumes and flowers. Although Fusarium oxysporum has a broad host range as a whole, a single strain can only infect one or a few plant species. It is divided into different physiological specialization types on different hosts. At present, there are more than 150 physiological specialization types reported. Some specialization types are further divided into different physiological races according to the specificity of different species. Most specializations contain 2 or more races, while a small part are monotypes. A certain physiological specialization may cause disease on a specific ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11C12R1/645
CPCC12Q1/6895
Inventor 杨腊英黄俊生郭立佳汪军刘磊梁昌聪王国芬陈平亚师雪茹
Owner ENVIRONMENT & PLANT PROTECTION INST CHINESE ACADEMY OF TROPICAL AGRI SCI