A transcription factor of root-knot nematode mgbtf3 and its application in disease control

A root-knot nematode, transcription factor technology, applied in application, nematicide, genetic engineering, etc., can solve the problem of undiscovered effector protein of transcription factor type

Active Publication Date: 2022-04-08
RICE RES INST GUANGDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In plant-parasitic nematodes, domestic and foreign studies have also reported that nematodes can perform transcriptional regulation on host cells during the process of parasitic plants. However, transcription factor-type effector proteins in plant-parasitic nematodes have not been found.

Method used

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  • A transcription factor of root-knot nematode mgbtf3 and its application in disease control
  • A transcription factor of root-knot nematode mgbtf3 and its application in disease control
  • A transcription factor of root-knot nematode mgbtf3 and its application in disease control

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 Cloning and sequence analysis of MgBTF3 gene

[0049] Based on the secretome of M. incognita and the genome of M. graminearum, the full-length cDNA sequence of MgBTF3 was amplified from M. graminearum through homologous cloning technology ( figure 1 ). The full-length cDNA sequence of MgBTF3 contains an open reading frame of 492bp, encoding 163aa, with a size of 18kDa. Using SignalP, it was found that the MgBTF3 protein does not contain a typical secretion signal peptide at the N-terminus. Further predicting SecretomeP 2.0 software by atypical secreted protein, the results showed that BTF3 may be an atypical signal peptide secreted protein. At the same time, TMHMM 2.0 online prediction of the transmembrane domain showed that MgBTF3 did not have a transmembrane domain.

Embodiment 2

[0050] Example 2 The MgBTF3 gene is specifically expressed in the head sensory organs of nematodes, and the expression is highest at the stage of feeding site formation

[0051] In this study, in situ hybridization was used to determine the expression position of MgBTF3 gene in Meloidogyne graminearum. The results showed that the sense strand probe of MgBTF3 had no obvious signal on the worm body (Fig. 2A). On the contrary, the antisense strand probe had obvious hybridization signal in the head sensilla of the 2nd instar larvae ( figure 2 B-2C), illustrating the site-specific expression of MgBTF3 in the head sensilla of M. graminearum.

[0052] The developmental expression of MgBTF3 gene was analyzed by real-time fluorescence quantitative qRT-PCR. In this experiment, five stages of the life cycle of root-knot nematode graminaceae were selected: eggs, 2nd instar larvae before infection (pre-J2s), 2nd instar worms in the parasitic stage (par-J2s), 3rd and 4th instar larvae (...

Embodiment 3

[0054] Example 3 The acquisition and specificity detection of MgBTF3 antiserum

[0055] In this study, the MgBTF3 immunogen protein ( image 3 A). The polyclonal antiserum of MgBTF3 was obtained by immunizing New Zealand white rabbits. The total protein of pre-J2s and healthy rice roots was extracted, and the specificity of MgBTF3 antiserum was verified by western blot. The results of Western blot showed that the MgBTF3 antiserum had a specific hybridization band of about 28kDa in the total protein of pre-J2s, which was 10kDa larger than the expected molecular weight of MgBTF3 protein (18kDa). In contrast, MgBTF3 antiserum showed no hybridization signal in the total protein of healthy rice roots. The comprehensive results showed that the MgBTF3 polyclonal antiserum could specifically recognize the MgBTF3 protein in nematodes, and had no non-specific signal with the total protein in rice roots. It is speculated that MgBTF3 may undergo protein modification in nematodes, lead...

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Abstract

The invention discloses a transcription factor MgBTF3 of root-knot nematode graminaceae and its application. MgBTF3 is the first transcription factor-type effector protein cloned from root-knot nematode. It is specifically expressed in the head sensilla, the secretory organ of nematode effector proteins, and is secreted into the host plant during nematode infection. Experiments have shown that reducing the transcription level of MgBTF3 during nematode infection will lead to a significant decrease in nematode infectivity. At the same time, overexpression of MgBTF3 in rice can significantly improve the infectivity of nematodes. Knocking out the endogenous OsBTF3 in the host rice, the nematode's ability to infect rice decreased, and the complementation experiment proved that MgBTF3 and rice OsBTF3 have similar functions. The research on MgBTF3 of the present invention reveals a new strategy for the root-knot nematode effector protein to suppress the host immune response: regulating the host immune system by simulating the host's endogenous protein. The root-knot nematode MgBTF3 and the corresponding rice endogenous OsBTF3 have important application value and prospects in the prevention and control of root-knot nematode graminearum.

Description

technical field [0001] The invention belongs to the technical field of plant disease control. More specifically, it relates to a transcription factor MgBTF3 of root-knot nematode graminaceae and its application in disease control. Background technique [0002] Root-knot nematode is an important class of plant pathogens. For example, Meloidogynegraminicola is an important parasitic nematode in rice roots, which causes serious economic losses to rice. [0003] Studies have shown that root-knot nematode effector proteins can be secreted into host tissues during the parasitic process of nematodes, thereby regulating the physiological metabolism and immune system of host cells to enhance the pathogenicity of nematodes. For example, Chinese patent CN108611352B discloses a translation elongation factor Mg-eEF1A of root-knot nematode graminearum, which can be secreted by root-knot nematode graminearum to activate the immune system of the host during the infection of plant parasitic...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/435C12N15/12C12N15/113C12N15/29C12N15/82A01N57/16A01P5/00A01H5/00A01H6/46
CPCC07K14/4354C07K14/415C12N15/113C12N15/8218C12N15/8285A01N57/16C12N2310/14
Inventor 陈建松李志文
Owner RICE RES INST GUANGDONG ACADEMY OF AGRI SCI
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