Molecular marker closely linked to salt tolerance of tomato and application thereof

A technology of salt tolerance and tomato, applied in the field of tomato breeding and molecular biology, can solve the problems of ion poisoning and lack of versatility, and achieve the effects of wide application, speeding up the breeding process, and improving the efficiency and accuracy of selection.

Active Publication Date: 2019-01-18
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Second, ion poisoning
Markers for salt tolerance are currently rare and, more importantly, not universal

Method used

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  • Molecular marker closely linked to salt tolerance of tomato and application thereof
  • Molecular marker closely linked to salt tolerance of tomato and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] A tomato salt-tolerant molecular marker STQ4 obtained by the following method:

[0031] 1. Group building

[0032] Salt-sensitive cultivated tomato TS-3 and salt-tolerant wild gooseberry tomato TS-21 are both resequenced tomato germplasm resources (Linet al.Nature Genetics2014,46:1220-1226), with TS-3 as the female parent, TS -21 is the male parent crossed to obtain F1, and then self-crossed to obtain F2, and the RIL population was constructed by the method of single son generation.

[0033] 2. Identification of salt tolerance

[0034] (1) The RIL6 population was planted in a 7×7cm black square pot containing the same amount of nutrient soil, and there were 2 replicates. The plants were grown in a greenhouse with a temperature of 25±2°C, a photoperiod of 16h light / 8h dark, a light intensity of 3500Lux, and normal fertilizer and water management.

[0035] (2) When the seedlings have 2 leaves and 1 heart, take a small amount of samples of each line of plants and freeze...

Embodiment 2

[0044] Verification of tomato salt-tolerant molecular marker STQ4:

[0045] Using the obtained salt-tolerant molecular marker primers, 30 parts of salt-sensitive materials and 30 parts of salt-tolerant materials screened in step 2 of Example 1 were used for salt-tolerance identification. The steps are as follows:

[0046] (1) Using tomato genomic DNA as a template, the concentration is 80-120ng / μl; molecular marker STQ4 is used for PCR amplification, the sequence of the forward primer is SEQ ID NO.1, and the sequence of the reverse primer is SEQ ID NO.2. PCR reaction system: the total volume is 15 μl, and the specific components are as follows: ddH 2 O 10.4μl, 10×PCR Buffer1.5μl, 1.2μldNTPs (2mmol / L), 0.08μl Taq DNA polymerase (5U / μl), 0.4μl forward primer (10ng / μl) (sequence SEQ ID NO.1), 0.4 μl reverse primer (10 ng / μl) (sequence SEQ ID NO.2) and 1.0 μl DNA template (100 ng / μl).

[0047] (2) The PCR reaction was carried out on the S1000 PCR machine produced by Bio-Rad Comp...

Embodiment 3

[0052] Application of tomato salt-tolerant molecular marker STQ4 in tomato salt-tolerant breeding:

[0053] 1) Using the method in Example 2, 204 tomato germplasms (including cultivated tomato and wild tomato) were identified by PCR, and 19 parts of materials (TS-7, TS-8, TS-13, TS-17, TS-18, TS-19, TS-20, TS-22, TS-56, TS-79, TS-80, TS-87, TS-116, TS-123, TS- 144, TS-164, TS-208, TS-226, TS-295).

[0054] 2) To identify the salt tolerance of the materials obtained above, at the same time, 5 copies of cultivated tomato (TS-131, TS-165, TS-166, TS-195, TS-201) and 4 copies of Gooseberry tomato (TS-16, TS-24, TS-77, TS-92) was used as a control. The specific steps are:

[0055] (1) The plants of each salt-tolerant material screened by molecular markers were planted in sprout trays, with 8 plants for each material, randomly distributed, and 3 replicates. The plants were grown in a greenhouse with a temperature of 25±2°C, a photoperiod of 16h light / 8h dark, a light intensity o...

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Abstract

The invention discloses a molecular marker closely linked to salt tolerance of tomato and application thereof. The RIL population was constructed by using salt-tolerant gooseberry tomato as male parent and salt-intolerant tomato as female parent. Chromosomal regions and candidate genes closely linked to salt tolerance were identified by high-throughput sequencing and recombinant plant analysis. Molecular markers were designed according to gene variations. Using this marker to identify RIL population, the coincidence rate was 96.7%. At the same time, the coincidence rate was 94.7% by using themarker to screen the natural population and identify the salt tolerance. Therefore, the present invention provides closely linked molecular markers for map-based cloning of tomato salt-tolerant genesand salt-tolerant breeding, and has important utilization value.

Description

technical field [0001] The invention belongs to the field of tomato breeding and molecular biology, more specifically relates to a molecular marker closely related to tomato salt tolerance, and also relates to a molecular marker closely related to tomato salt tolerance in tomato salt tolerance breeding application. Background technique [0002] Soil salinization is one of the most serious abiotic stresses on plants, affecting more than 1 billion hectares of land in more than 100 countries, seriously affecting the yield and quality of crops and vegetables. At present, global climate change has further led to the continuous expansion of drought and saline soil, and the development of facility agriculture has led to the increase of secondary salinization of soil year by year. At present, the global soil salinization area is increasing at a rate of 10% every year. According to statistics, there are about 36.9 million hectares of salinized land in China, accounting for about 4....

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 欧阳波赵钢军刘敏敏卢永恩李汉霞
Owner HUAZHONG AGRI UNIV
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