Sweet potato stem nematode resistance related protein IbPIF1 as well as coding gene and application thereof

A resistance-related, stem nematode technology, applied in application, genetic engineering, plant genetic improvement, etc., to achieve the effects of improved disease resistance, broad application space and market prospects

Active Publication Date: 2022-05-27
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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  • Sweet potato stem nematode resistance related protein IbPIF1 as well as coding gene and application thereof
  • Sweet potato stem nematode resistance related protein IbPIF1 as well as coding gene and application thereof
  • Sweet potato stem nematode resistance related protein IbPIF1 as well as coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Example 1. Obtaining the protein IbPIF1 and its encoding gene related to the improvement of sweet potato stem nematode disease

[0071] 1. Extraction of total RNA from sweet potato

[0072] Take 0.1 g of young leaves of sweet potato variety Lushu No. 3 and grind them into powder in liquid nitrogen, add them to a 2 mL centrifuge tube, and use TIANGEN's RNAprep pure plant total RNA extraction kit (catalog number: DP432) to extract total sweet potato RNA, kit Included: Lysis Buffer RL, Deproteinization Buffer RW1, Wash Buffer RW, RNase-Free ddH 2 O, RNase-Free adsorption column CR3, RNase-Free filter column CS, DNase I, buffer RDD, RNase-Free centrifuge tube, RNase-Free collection tube.

[0073] Take 1 μL of the extracted total RNA to detect its integrity by 1.2% agarose gel electrophoresis, and take another 2 μL of the extracted total RNA to dilute to 500 μL, and use a UV spectrophotometer to detect its quality (OD). 260 ) and purity (OD 260 / OD 280 ), the total RNA e...

Embodiment 2

[0080] Example 2. Application of IbPIF1 protein in improving sweet potato D. nematode resistance

[0081] 1. Preparation of IbPIF1-transformed sweet potato

[0082] 1. Construction of plant expression vector

[0083] 1) According to the coding sequence of the sweet potato IbPIF1 gene, design and amplify the primer sequence of the complete coding sequence, and respectively introduce NcoI and PmlI restriction sites in the forward primer (primer 3) and reverse primer (primer 4), the primers The sequence is as follows:

[0084] Primer 3: 5'- CATGCCATGG ATGGATGAGGACTTCAATCTTCCT-3' (the underlined part is the NcoI restriction site);

[0085] Primer 4: 5'- GCCACGTG TCAACGAGATTGATGATTCCCTG-3' (the underlined part is the PmlI restriction site).

[0086] 2) Take the DNA fragment shown in the artificially synthesized SEQ ID No.2 as template, adopt primer 3 and primer 4 to carry out PCR amplification, obtain PCR product, then PCR product is connected to pMD19-T carrier (purchased f...

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Abstract

The invention discloses a sweet potato stem nematode resistance related protein IbPIF1 as well as a coding gene and application thereof. The IbPIF1 is a protein as shown in a1) or a2) or a3) or a4): a1) a protein as shown in a sequence 2; a2) a fusion protein obtained by connecting a tag to the N terminal or/and C terminal of the protein as shown in the sequence 2; a3) a stem nematode disease resistance related protein obtained by substitution and/or deletion and/or addition of one or more amino acid residues of the amino acid sequence shown in the sequence 2; a4) a protein which has 90% identity with the amino acid sequence shown in the sequence 2, is derived from sweet potatoes and is related to stem nematode disease resistance. The coding gene of the IbPIF1 protein is introduced into the sweet potato to obtain a transgenic plant, and the transgenic plant is subjected to stem nematode infection to find that the resistance of the transgenic plant is enhanced. It is shown that the IbPIF1 protein plays an important role in the plant disease resistance process and has a wide application prospect.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a protein IbPIF1 related to sweet potato stem nematode disease resistance and its encoding gene and application. Background technique [0002] Sweet potato is an important food, feed, industrial raw material and new energy plant, and its status is particularly important. my country is the largest sweet potato producer in the world, with an annual planting area of ​​3.291 million hectares 2 , accounting for 38.1% of the world's total planting area, and the annual output accounts for 67.3% of the world's total output. With the rapid growth of the world's population and the reduction of arable land, the security of food production has been seriously threatened. In agricultural production, pests and diseases are always one of the serious threats to crop production, affecting the yield and quality of crops, and sweet potato is no exception. Cultivating sweet potato varieties with good ...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/00
CPCC07K14/415C12N15/8285C12N15/8216Y02A40/146
Inventor 翟红刘庆昌何绍贞高少培张欢霍进喜聂楠
Owner CHINA AGRI UNIV
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