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MicroRNA167a for regulation related with induction of plant male sterility and application of microRNA167a

A technology for male sterility and plant induction, applied in the direction of DNA/RNA fragments, applications, plant products, etc., can solve the problems of wheat miR167, etc., and achieve the effects of improving breeding models, simplifying breeding procedures, and accelerating applications

Active Publication Date: 2017-05-10
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the function of miR167 in wheat

Method used

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  • MicroRNA167a for regulation related with induction of plant male sterility and application of microRNA167a
  • MicroRNA167a for regulation related with induction of plant male sterility and application of microRNA167a
  • MicroRNA167a for regulation related with induction of plant male sterility and application of microRNA167a

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: cDNA cloning of tae-pre-miR167a related to light-temperature-sensitive male sterility in wheat

[0033] The anthers of wheat BS366 were taken, the total RNA of the anthers was extracted by Trizol method, and cDNA was obtained by reverse transcription with superscript II (purchased from Invitrogen Company) reverse transcriptase. Using the specific primers P1 and P2 of tae-pre-miR167a, the cDNA obtained by reverse transcription was used as a template for PCR amplification, and the amplified products were as follows: figure 1 shown. The sequences of primers P1 and P2 are as follows:

[0034] P1: 5'-CTGCCCAAGGGAACGAGTGAA-3'

[0035] P2: 5'-AGAAGTAGTTGTGCCCAACGGC-3'

[0036] The PCR product was detected by 1% agarose gel electrophoresis, and a band with a length of about 120 bp was obtained, and the fragment was recovered with an agarose gel recovery kit (TIANGEN). The recovered fragment was ligated with pGEM-T Easy (purchased from Promega Company), and with r...

Embodiment 2

[0037] Embodiment 2: Cultivate male sterile transgenic plants with Tae-pre-miR167a gene

[0038] 1. Construction of recombinant expression vector

[0039] Construction of recombinant expression vector of tae-pre-miR167a dicot

[0040] Using the cDNA obtained by reverse transcription of the total microRNA of wheat BS366 as a template, PCR amplification was performed with specific primers containing XbaI and SmaI linker sequences; then the PCR product was digested and recovered, and the digested product was inserted into the vector pBI121 to obtain the recombinant vector pBI121 -tae-pre-miR167a.

[0041] The primer sequences are as follows:

[0042] tae-pre-miR167a[XbaI]: 5'–GC TCTAGA CTGCCCAAGGGAACGAGTGAA-3'

[0043] tae-pre-miR167a[SmaI]: 5'–TCC CCCGGG AGAAGTAGTTGTGCCCAACGGC-3'

[0044] 2. Acquisition and identification of transgenic Arabidopsis

[0045] 1) Obtaining transgenic Arabidopsis

[0046] The recombinant expression vector pBI121-tae-pre-miR167a constructed ab...

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Abstract

The invention relates to the field of genetic engineering of plants, in particular to microRNA167a for regulation related with induction of plant male sterility and an application of microRNA167a. The mature sequence of microRNA167a is shown in SEQ ID NO.1, and the precursor sequence of microRNA167a is shown in SEQ ID NO.2. Tae-pre-miR167a deriving from wheat can induce sterility of pollen in stamens of arabidopsis, so that fruitlessness occurs to fruit pods.

Description

technical field [0001] The invention relates to the field of plant genetic engineering, in particular to microRNA167a for inducing plant male sterility-related regulation and its application. Background technique [0002] In modern agricultural production, the use of heterosis is considered to be the main way to greatly improve the wheat production level in the future. At present, breeders have used transgenic technology to transfer some genes related to fertility regulation into plants, and obtained partially or even completely male sterile plants. [0003] MicroRNAs (miRNAs) are a class of 21-25nt non-coding small RNA molecules that can bind to the 3' untranslated region of mRNA molecules through base pairing to degrade mRNA or inhibit the transcription of target genes. Regarding the report of miR167 in plant research, it was found in Arabidopsis that only the overexpression of miR167a caused curly flowers, short tidbits, and inhibited flower development; in rice, it was ...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/82A01H5/00
CPCC12N15/113C12N15/8218C12N15/8289C12N2310/141
Inventor 张立平赵昌平段文静王玉昆苑少华白建芳王鹏苑国良张风廷王娜
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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