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Application and application method of potato x virus in inducing embryogermination of tomato seeds

An application method, potato technology, applied in the biological field, can solve the problems of tomato fruit not ripening, lack of evidence, etc.

Active Publication Date: 2021-07-20
HANGZHOU NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, although there is sporadic evidence that epigenetics may be involved in the regulation of seed dormancy and / or germination, however, whether epigenetics is involved or how to control plant embryogenesis is lacking, and further research is needed
[0004] tomato Colourless non-ripening ( Cnr ) is a naturally occurring epigenetic mutant in which, LeSPL-CNR The promoter of the gene contains a highly methylated 286bp region, and hypermethylation inhibits LeSPL-CNR Gene expression that causes tomato fruit to fail to ripen

Method used

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  • Application and application method of potato x virus in inducing embryogermination of tomato seeds
  • Application and application method of potato x virus in inducing embryogermination of tomato seeds
  • Application and application method of potato x virus in inducing embryogermination of tomato seeds

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1: Using VIGS Research SlMET1 Whether to participate in tomato seed germination

[0057] 1. Using Trizol, total RNA was extracted from tomato leaves according to the method provided by the manufacturer. Using this as a template, the first-strand cDNA was synthesized using reverse transcriptase.

[0058] 2. Design primers according to the sequence of the tomato SlMET1 gene registered in the NCBI database, and entrust Shanghai Yingjun Company to synthesize them. The primer sequence information is shown in Table 1:

[0059] Table 1

[0060] .

[0061] 3. In the PCR reaction, the first-strand cDNA is used as a template, and primer 1 and primer 2 are used as primers. Take a 0.5ml PCR thin-walled tube and add the following reagents one by one: template DNA 2μl, 10mmol / l dNTP mixture 0.5μl, 20μmol / l upstream and downstream primers 0.5μl, 5× reaction buffer 4μl, 2.5U / μl PrimerSTAR HS DNA polymerase 0.1 μl, add double-distilled water to a final volume of 20 μl; PCR...

Embodiment 2

[0092] Example 2: Study using VIGS SlCMT4 Whether to participate in tomato seed germination

[0093] 1. Synthesize Arabidopsis first-strand cDNA according to the method described in Example 1;

[0094] 2. According to the tomato registered in the NCBI database SlCMT4 Primers were designed for the sequence of the gene and synthesized by Shanghai Yingjun Company. See Table 2 for the sequence information of the primers:

[0095] Table 2

[0096] ;

[0097] 3. Obtained according to step 3 of the method in Example 1 SlCMT4 PCR product fragments, the primers used are primer 3 and primer 4;

[0098] 4, construct PVX / SlCMT4 according to embodiment 1 method step 4, as figure 1 shown;

[0099] 5. Prepare linearized DNA according to step 5 of the method in Example 1;

[0100] 6. Perform in vitro transcription according to step 6 of the method in Example 1;

[0101] 7. Carry out the virus inoculation of Nicotiana benthamiana according to embodiment 1 method step 7;

[0102...

Embodiment 3

[0103] Example 3: Study using VIGS SINCED Whether to participate in tomato seed germination

[0104] 1. Synthesize Arabidopsis first-strand cDNA according to the method described in Example 1;

[0105] 2. According to the tomato registered in the NCBI database SINCED Primers were designed for the sequence of the gene and synthesized by Shanghai Yingjun Company. The sequence information of the primers is shown in Table 3:

[0106] table 3

[0107] ;

[0108] 3. Obtained according to step 3 of the method in Example 1 SINCED PCR product fragments, the primers used are primer 5 and primer 6;

[0109] 4, construct PVX / SINCED according to embodiment 1 method step 4, as figure 1 shown;

[0110] 5. Prepare linearized DNA according to step 5 of the method in Example 1;

[0111] 6. Perform in vitro transcription according to step 6 of the method in Example 1;

[0112] 7. Carry out the virus inoculation of Nicotiana benthamiana according to embodiment 1 method step 7;

[...

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Abstract

The application and application method of potato X virus in inducing tomato seed germination belong to the field of biotechnology. On the one hand, the invention discloses the application of potato X virus in inducing tomato seed germination, and on the other hand, discloses the application method of potato X virus in inducing tomato seed embryo germination. The method of the present invention does not need cumbersome transformation and transgene screening processes, and the process of studying tomato seed embryo germination only takes about 2 months, which can efficiently and quickly research tomato seed embryo germination; and by utilizing the potato virus X vector to study tomato seed embryo germination, It is confirmed that the epigenetic mechanism is involved in the regulation of plant seed germination.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application and application method of potato X virus in inducing embryo germination of tomato seeds. Background technique [0002] Vivipary, also known as post-harvest sprouting, is a survival adaptation mechanism that some plants have evolved to resist extreme environments; but for agricultural and fruit crops, vivipary will reduce yield, It will affect the quality of grains and fruits, and cause great harm to global food production and safety. [0003] In food and fruit crops, such as corn, wheat, rice and tomato, some studies have been carried out on fetal germination, and related genetic mechanisms have been discovered and some genes involved in the regulation of fetal germination have been revealed. At the same time, although there is sporadic evidence that epigenetics may be involved in the regulation of seed dormancy and / or germination, whether epigenetics is ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82A01H5/10A01H6/82
CPCC12N15/8203C12N15/8267
Inventor 洪益国陈微微
Owner HANGZHOU NORMAL UNIVERSITY
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