Litchi disease-resistant gene LcLTP as well as encoding protein and application thereof

A disease-resistant gene and protein-encoded technology, applied in the field of molecular plant pathology, can solve problems affecting the occurrence of diseases, and achieve the effect of enhanced expression and reduced disease spots

Active Publication Date: 2021-07-09
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The genome sequencing of P. litchi and P. litchi was completed late, so molecular biology research started relatively late, but now a research system for the interaction between ...

Method used

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  • Litchi disease-resistant gene LcLTP as well as encoding protein and application thereof
  • Litchi disease-resistant gene LcLTP as well as encoding protein and application thereof
  • Litchi disease-resistant gene LcLTP as well as encoding protein and application thereof

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Experimental program
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Effect test

Embodiment 1

[0034] Embodiment 1: Cloning and acquisition of LcLTP gene

[0035] (1) Through the analysis of the litchi genome, it was predicted that the litchi may encode the LcLTP gene. We designed primers (upstream primer LcLTP-F and downstream primer LcLTP-R), and successfully cloned the LcLTP gene in the litchi cDNA. Its nucleotide sequence is as follows: Shown in SEQ ID NO:1, the amino acid sequence of its encoded protein is shown in SEQ ID NO:2:

[0036] LcLTP gene (SEQ ID NO: 1; base composition: 80A, 87T, 109G, 66C):

[0037] ATGGGAAGCACCAAGGGAACATCACTAGGTTTTATGGTATTAGTAGTTGTAGCTGTTGTGGGGAAGTGGGAGGTGAAGATGGCTGGTGCAGAACTTAGTGCAGCCCAGTGCAAGGAAGAGAGGAGAATTGGGCTGAATGAGTGCAAGCCAGTGGTGTATGGGAAGCTTCCGTCGCCGTCGTGCTGTGAGCGTGTAAGGGTGAGTCATGTTGAATGTGTGTGCCCTGTCATTACACCTAAGTTGGCTGCTCTTATTGATCTCAACCGTGCCATCCGCCTCATCGAAGGCTGCGGTAGAAGAGTCCCTCGCCACTTCAAGTGTGGAAGTATCACAACTCCTTGA。

[0038] LcLTP protein (SEQ ID NO:2):

[0039] MGSTKGTSLGFMVLVVVAVGKWEVKMAGAELSAAQCKEERRIGLNECKPVVYGKLPSPSCCERVRVSHVE...

Embodiment 2

[0047] Example 2: Transient expression of LcLTP in tobacco

[0048] (1) Tobacco transient expression vector construction:

[0049] ① The target fragment is connected to the carrier: the amplified LcLTP target fragment is connected to the PVX vector with homologous recombinase (linkage site: SmaI) to obtain a recombinant plasmid containing the sequence of SEQ ID NO: 1; the GFP sequence (SEQ ID NO: 3) Use homologous recombinase to connect with PVX vector (connection site: SmaI), obtain recombinant plasmid PVX::GFP containing GFP green fluorescent protein, after breeding in Escherichia coli JM109 strain, extract the plasmid, and recombined plasmid PVX::GFP and the recombinant plasmid containing the sequence of SEQ ID NO:1 were transformed into Agrobacterium GV3101.

[0050] GFP gene (SEQ ID NO: 3):

[0051] ATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAGCTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTACGGCAAGCTGACCCTGAAGTTCATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGC...

Embodiment 3

[0055] Embodiment 3: Zoospores infect litchi leaves and branches

[0056] 1. Cultivation of Pythophthora lychee

[0057] The hyphae blocks of Pyrosophthora lychee strains preserved in the laboratory were inoculated on the carrot medium, and placed in the culture room at 27°C for one week.

[0058] 2. Preparation of zoospore suspension

[0059] Pour sterilized water on the Lychee downy mildew plate, use a brush to collect the litchi downy Phytophthora mycelium through the filter in a 50ml centrifuge tube, place it at 18°C ​​for 2 hours until it releases zoospores, when the sporangia are observed under the microscope After releasing the zoospores, the zoospore suspension was collected by filtration.

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Abstract

The invention discloses a litchi disease-resistant gene LcLTP as well as an encoding protein and application thereof. A litchi disease resistance related gene LcLTP is separated from litchis, the full-length cDNA of the gene is 342bp, and the nucleotide sequence of the gene is as shown in SEQ ID NO: 1; the protein coded by the gene is a lipid transfer protein with the full length consisting of 113 amino acids, and the amino acid sequence is shown as SEQ ID NO: 2. Transcription pattern analysis and plant expression analysis find that the LcLTP gene can improve the resistance of plants to phytophthora, so that the litchi disease-resistant gene LcLTP can be applied to the aspect of improving the disease resistance of the plants or breeding disease-resistant plants.

Description

technical field [0001] The invention belongs to the technical field of molecular plant pathology, and in particular relates to a litchi disease-resistant gene LcLTP, its encoded protein and its application. Background technique [0002] Litchi belongs to Sapindaceae, evergreen trees, and is an important tropical fruit in southern my country, with important economic value; it is often distributed in Guangdong, Fujian, Guangxi, Sichuan and other places in China, and is called "Southern Country" together with banana, pineapple and longan. Four major fruits". [0003] Litchi frost blight and anthracnose are the two most serious diseases on litchi; among them, litchi frost blight is caused by Peronophythora litchii Chen ex Ko et al, which usually infects hosts through sporangia and zoospores . Litchi frost blight can damage litchi leaves, flower spikes, branches, and fruits, and produce brown lesions. When the humidity is high, it can produce a white frosty mold layer. The dryn...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/82A01H6/00
CPCC07K14/415C12N15/8282
Inventor 孔广辉冯迪南李雯刘家琪黄伟雄司徒俊键习平根李敏慧姜子德
Owner SOUTH CHINA AGRI UNIV
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