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Molecular marker primer of disease-resistant gene ty-5 of tomato yellow leaf curl disease and application of molecular marker primer

A tomato yellow leaf curl and molecular marker technology, applied in the field of tomato yellow leaf curl disease resistance gene ty-5 molecular marker primers, can solve the problems of cumbersome operation, high experimental cost, unclear linkage distance, etc., and achieve improvement Economic benefits, convenient operation, and the effect of ensuring smooth development

Active Publication Date: 2015-12-23
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, in the study of ty-5 linkage markers, only one CAPS marker SlNAC1 linkage has been identified, and the linkage distance is still unclear; (see HuttonSF, ScottJW, SchusterDJ. Recessive Resistance to Tomatoyellowleaf curlvirus from the TomatoCultivarTykingIsLocatedintheSameRegionasTy-5onChromosome4. Hortscience, 2012, 47(3): 724-3) , and the detection of this type of marker requires enzyme digestion of the PCR product, which is costly and cumbersome to operate.

Method used

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  • Molecular marker primer of disease-resistant gene ty-5 of tomato yellow leaf curl disease and application of molecular marker primer
  • Molecular marker primer of disease-resistant gene ty-5 of tomato yellow leaf curl disease and application of molecular marker primer
  • Molecular marker primer of disease-resistant gene ty-5 of tomato yellow leaf curl disease and application of molecular marker primer

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Experimental program
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Effect test

Embodiment 1

[0049] The molecular marker of the tomato yellow leaf curl resistance gene ty-5 is an InDel marker type, which utilizes the following molecular marker primers of the tomato yellow leaf curl resistance gene ty-5:

[0050] The upstream primer from 5' end to 3' end is: TAACAAAGCCCTCAAAGC

[0051] The 5' end to the 3' end of the downstream primer is: GTCTCCGAAACGTAATCC;

[0052] A co-dominant DNA molecular marker closely linked to ty-5 obtained through PCR amplification;

[0053] The PCR reaction system is: a total of 10 μL reaction system, 10 ng / μL template DNA 1.0 μL; 4 pmmol / L molecular marker primer 1.0 μL, 10×PCRBuffer 1.0 μL, 25 mmol / LMgCl 2 1.0μL, 10mmol / LdNTPs0.25μL, 5U / μLrTaqDNAPolymerase0.1μL, ddH 2 O5.65 μL;

[0054] The PCR reaction amplification program is: 94°C pre-denaturation for 5 minutes; each cycle of 94°C denaturation for 30 s, 48°C annealing for 30 s, 72°C extension for 30 seconds, a total of 38 cycles, and finally 72°C extension for 7 min;

[0055] The nu...

Embodiment 2

[0057] The molecular marker of the tomato yellow leaf curl disease resistance gene ty-5 is an InDel marker type, and is a molecular marker primer utilizing the above tomato yellow leaf curl disease resistance gene ty-5:

[0058] The upstream primer from 5' end to 3' end is: TAACAAAGCCCTCAAAGC

[0059] The 5' end to the 3' end of the downstream primer is: GTCTCCGAAACGTAATCC;

[0060] A co-dominant DNA molecular marker closely linked to ty-5 obtained through PCR amplification;

[0061] The PCR reaction system is: a total of 10 μL reaction system, 10 ng / μL template DNA 1.0 μL; 4 pmmol / L molecular marker primer 1.0 μL, 10×PCRBuffer 1.0 μL, 25 mmol / LMgCl 2 1.0μL, 10mmol / LdNTPs0.25μL, 5U / μLrTaqDNAPolymerase0.1μL, ddH 2 O5.65 μL;

[0062] The PCR reaction amplification program is: 94°C pre-denaturation for 5 minutes; each cycle of 94°C denaturation for 30 s, 48°C annealing for 30 s, 72°C extension for 30 seconds, a total of 38 cycles, and finally 72°C extension for 7 min;

[0063...

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Abstract

The invention discloses a molecular marker primer of a disease-resistant gene ty-5 of the tomato yellow leaf curl disease and application of the molecular marker primer. Sequences of the molecular marker primer are TAACAAAGCCCTCAAAGC from 5' terminal to 3' terminal of a forward primer and GTCTCCGAAACGTAATCC from 5' terminal to 3' terminal of a reverse primer. A molecular marker is of an InDel marker type, a nucleotide sequence of the molecular marker is SEQ ID No.3 or SEQ ID No.4. The InDel molecular marker linked with the gene is developed by a tomato material AVTO 1227 containing the gene ty-5 and is used for molecular marker-assisted selection. The molecular marker which is the InDel marker can be used for pleomorphic analysis without being subjected to enzyme digestion, and accordingly, convenience and simplicity in operation and low experiment cost are achieved.

Description

technical field [0001] The invention belongs to the field of biotechnology, relates to the technical field of molecular markers, in particular to a molecular marker primer of tomato yellow leaf curl disease resistance gene ty-5 and an application thereof. Background technique [0002] Tomato (Solanum lycopersicum) is an important vegetable crop in the world, which plays a huge role in promoting agricultural economic development and increasing farmers' income. The planting process is susceptible to various diseases, resulting in a decrease in yield, so it is very important to breed disease-resistant varieties. [0003] Tomato yellow leaf curl disease (TomatoyellowleafcurldiseaseTYLCD) is an important disease of tomato. Lice (Bemisiatabaci) spread. The disease was first reported in Israel. With the deepening of international trade, TYLCV gradually expanded to the global scope. In the 1990s, the disease was introduced into my country. Now it has spread all over the main tomat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/70C12Q1/68
Inventor 王银磊
Owner JIANGSU ACAD OF AGRI SCI
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