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Gene capable of reducing seed formation and application thereof in cultivatingseedless plants

A gene and plant technology, applied in the field of plant tissue culture, can solve the problems of long cultivation time and high cost, and achieve the effect of reducing seed formation and production

Pending Publication Date: 2019-06-14
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Cultivate seedless melons and fruits by triploid or induced parthenocarpy, which takes a long time and high cost

Method used

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  • Gene capable of reducing seed formation and application thereof in cultivatingseedless plants
  • Gene capable of reducing seed formation and application thereof in cultivatingseedless plants
  • Gene capable of reducing seed formation and application thereof in cultivatingseedless plants

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0048] The sixth embodiment of this specification provides a gene preparation method as described in the first embodiment, comprising the following steps:

[0049] Extract the RNA in the seedless grape ovule, and use the extracted RNA as a template to perform reverse transcription to obtain cDNA;

[0050] Using the cDNA as a template, the sequence shown in SEQ ID NO: 3 as an upstream primer, and the sequence shown in SEQ ID NO: 4 as a downstream primer are amplified to obtain the gene.

[0051] In one example, the extraction of RNA in seedless grape ovules, and using the extracted RNA as a template, reverse transcription to obtain cDNA, including:

[0052] The RNA of the ovules of 20 days after anthesis, 30 days after anthesis, 40 days after anthesis, and 50 days after anthesis were respectively extracted, and reverse-transcribed respectively to obtain 20 days after anthesis, 30 days after anthesis, 40 days after anthesis, and 40 days after anthesis. After 50 days ovule cDNA;...

Embodiment 1

[0062] Embodiment 1, the extraction of seedless white grape ovule RNA

[0063] Observe the flowering time of European grapes Pinot Noir and Pinot Noir in the Yangling region of Shaanxi, China. When 80% of the buds on the whole cluster of Pinot Noir and Pinot Noir are open, mark the bagging. Samples were taken every 10 days from 20 days after flowering until 50 days after flowering. Respectively named 20DAF, 30DAF, 40DAF, 50DAF. The collected grape samples were placed in an ice box and brought back to the laboratory for embryo picking. The plucked embryos were quickly frozen in liquid nitrogen and stored in a -80°C refrigerator. Seedless white grape ovule RNA was extracted according to the instructions in the Plant Total RNA Extraction Kit (Plant RNA Kit, OMEGA), and the specific test steps were as follows:

[0064] (1) Quick-freeze the seedless white grape ovules in liquid nitrogen, and fully grind them to a powder state;

[0065] (2) Mix RCL Buffer and β-mercaptoethanol, ...

Embodiment 2

[0083] Example 2. Reverse transcription of cDNA from seedless white grape ovule RNA.

[0084] The reverse transcription process was carried out according to the instructions in Novizan Reverse Transcription Kit (VazymE), and the steps were as follows:

[0085] (1) Remove genomic DNA, and prepare the following mixture in an RNase-free PCR tube:

[0086]

[0087] Gently blow and mix with a pipette, 42°C for 2min.

[0088] (2) Prepare the reverse transcription reaction system:

[0089] Add 5×HiScriptⅡqRT SuperMixⅡ directly to the reaction tube in step 1

[0090]

[0091] Mix by gently pipetting.

[0092] (3) Carry out reverse transcription reaction:

[0093]

[0094] The reaction product is cDNA, which can be used immediately for qRT-PCR reaction, or stored in a -80°C refrigerator.

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Abstract

The embodiment of the invention provides a gene capable of effectively reducing seed formation and application thereof in cultivatingseedless plants. Expression of the gene in the plants can effectively reduce seed formation; the gene is the following DNA fragments of (1) or (2); (1) is the DNA fragment with the base sequence as shown in SEQIDNO:1, and (2) is the DNA fragment which has at least 80% homology with the DNA fragment in (1) and has thefunction of the DNA fragment in (1).

Description

technical field [0001] One or more embodiments of this specification relate to the technical field of plant tissue culture, in particular to a gene capable of effectively reducing seed formation and its application in cultivating seedless plants. Background technique [0002] Seedless breeding is an important direction of melon and fruit breeding, and many seedless melons and fruits have been put on the market now, such as seedless watermelons, seedless grapes, and seedless kiwis. At present, most of the seedless melons are obtained by breeding triploid or inducing parthenocarpy. [0003] Cultivating seedless melons and fruits by triploids or inducing parthenocarpy requires a long cultivation time and high cost. Contents of the invention [0004] One or more embodiments of this specification describe a gene capable of effectively reducing seed formation and its application in breeding seedless plants. [0005] According to a first aspect, an isolated gene is provided, th...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82A01H5/00A01H6/88A01H6/82
Inventor 徐炎张晨焦韵桐李志谦郝新意
Owner NORTHWEST A & F UNIV
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