Paeonia suffruticosa Andr. gene IKU2 preparation method

A technology of peony and gene, applied in the field of bioengineering, can solve problems such as unreported, and achieve the effects of improving transformation efficiency, increasing yield, and increasing plant yield

Active Publication Date: 2019-01-04
DALIAN NATIONALITIES UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the functional research of the IKU2 gene has only been verified in model plants such as Arabidopsis thaliana, and the function of the IKU2 gene in woody oil crops, especially the purple spot peony, has not been reported yet.

Method used

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  • Paeonia suffruticosa Andr. gene IKU2 preparation method
  • Paeonia suffruticosa Andr. gene IKU2 preparation method

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Embodiment 1

[0046] 1. Obtaining the IKU2 gene cDNA from the seeds of Peony

[0047] Collect seeds from the four developmental periods (June 20, July 7, July 17, and July 27) of the purple spotted peony base of Jinshitan campus of Dalian Nationalities University, mark them, and immediately freeze them with liquid nitrogen, and then freeze the seeds Grind with liquid nitrogen, and extract the total RNA of Paeonia edulis seeds after improving the method recommended by the column-type plant total RNA extraction and purification kit of Shanghai Shenggong Company. Using the extracted RNA as a template for reverse transcription reaction, the steps are as follows:

[0048] ① Prepare the following mixture in a PCR tube, and the reaction system is as follows:

[0049] Template RNA (300ng / μL) 2.0μL

[0050] Oligo(dT)18 Primer(50μM) 2.0μL

[0051] RNase free H2O 10.0μL

[0052] Total 14.0μL

[0053] ②Gently mix and centrifuge, react at 70°C for 10 minutes, and quickly place on ice to cool for 5 minutes;

[0054...

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Abstract

The invention relates to a Paeonia suffruticosa Andr. gene IKU2 preparation method and belongs to the technical field of bioengineering. The Paeonia suffruticosa Andr. gene IKU2 preparation method includes (1), extracting of total RNA of seed of Paeonia suffruticosa Andr.; (2), synthesis of cDNA first chain; (3), amplifying of a conserved region of the Paeonia suffruticosa Andr. gene IKU2; (4), recovering and purifying of PCR amplification product; (5), connecting of a target fragment and a cloning vector pMD 19-T; (6), preparing of colibacillus DH5 alpha competent cell; (7), converting of connecting products; (8), extracting of recombinant plasmid; (9), sequencing. According to the method, the gene IKU2 capable of regulating and controlling of size of seed is screened from the transcriptome library at different development stages of the seed of Paeonia suffruticosa Andr., and the molecular basis is provided for increasing the seed yield of Paeonia suffruticosa Andr. through the genetic engineering technology.

Description

Technical field [0001] The present invention belongs to the technical field of bioengineering, and specifically relates to a method for preparing the IKU2 gene of Paeonia edulis. Background technique [0002] Peony is a small perennial deciduous shrub of Paeoniaceae (Oaeonia). The development and utilization of woody oils can effectively alleviate the current situation of edible oil shortage in my country. In March 2011, the Supervision Bureau of the Ministry of Health of the People’s Republic of China publicly approved peony seed oil as a new product based on the provisions of the Food Safety Law and the review by the New Resources Food Review Expert Committee. Resource food, peony seed oil is designated as a new type of healthy woody edible oil. At present, China's large-scale demonstration planting of oil peony varieties mainly include Fengdan (P.ostii) suitable for southern climate and P.suffruticosa Andr.var.papaveracea suitable for northern climate. But its smaller seeds (...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10A01H5/10A01H6/00
CPCC07K14/415C12N15/8267
Inventor 丁健韩平阮成江闫蕊刘凌悦吴波董旭
Owner DALIAN NATIONALITIES UNIVERSITY
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