Paulownia BTB gene family and application thereof in biological and abiotic stress

Abstract

The invention discloses a paulownia BTB gene family and an application thereof in biological and abiotic stress, the paulownia BTB gene family comprises PfBT1-PfBT62, the paulownia BTB gene has an important defense effect on biological stress, the plant disease resistance is improved by overexpressing the paulownia BTB gene in a target plant, and under the stress of illness and health, PfBTB12, PfBTB14 and PfBTB44 are significantly up-regulated, and PfBTB3, PfBTB16, PfBTB19 and PfBTB36 are significantly down-regulated. By obtaining the related genes for resisting the occurrence of the paulownia arbuscular disease, the disease resistance rate of paulownia is reduced, and an important guiding effect is achieved for follow-up culture of disease-resistant paulownia varieties.

Description

Technical Field

[0001] The invention relates to the biological field, in particular to a Paulownia BTB gene family and applications thereof in biotic and abiotic stresses. Background Art

[0002] The BTB domain is a protein-protein interaction motif found in eukaryotes. Many proteins containing the BTB domain function as substrate-specific adaptors in Cullin3de-based E3 ubiquitin ligases, and plant BTB genes regulate biotic and abiotic stress responses.

[0003] Paulownia is an important deciduous tree. Due to its fast growth and excellent wood quality, it is often used in construction, furniture, and musical instruments, helping to alleviate the shortage of wood. Its flowers, leaves, fruits, and bark can be used as medicine and have high medicinal value. Paulownia has strong adaptability and resistance, and can improve soil, manage saline-alkali land, and improve the ecological environment. However, Paulownia trees are susceptible to phytoplasma infection, leading to the occurrence of Paulownia witches' broom disease.

[0004] Susceptibility to phytoplasma infection leads to axillary bud clusters, plant dwarfing, yellowing of leaves and other symptoms, resulting in slow growth or even death of large trees, seriously affecting the yield and economic benefits of Paulownia. This infection causes excessive growth of axillary buds and dwarfing of plants, increases tree mortality, slows tree growth, and seriously affects the growth of Paulownia production. Therefore, it is of great significance to identify genes related to resistance to Paulownia witches' broom. Summary of the invention

[0005] In order to solve the above problems, the present invention proposes a Paulownia BTB gene family, which includes PfBT1-PfBT62. The Paulownia BTB gene has an important defense effect on biological stress.

[0006] The invention discloses a paulownia BTB protein containing a paulownia BTB gene. The paulownia BTB protein comprises a BTB structural domain and an ankyrin repeat sequence and has both transcription activation and transcription inhibition effects.

[0007] The application of the Paulownia BTB gene family in biotic and abiotic stresses is to improve plant disease resistance by overexpressing the Paulownia BTB gene in target plants. Under diseased and healthy stresses, PfBTB12, PfBTB14 and PfBTB44 are significantly upregulated, and PfBTB3, PfBTB16, PfBTB19 and PfBTB36 are significantly downregulated.

[0008] A method for identifying a Paulownia BTB gene, comprising:

[0009] S1. Obtaining BTB genes in Paulownia: Download the Paulownia genome from NCBI, use HMM search software, and use the seed file of the BTB domain to search for the inferred BTB genes in the Paulownia genome database;

[0010] S2. Phylogenetic analysis of Paulownia BTB protein: The BTB protein sequence of Arabidopsis thaliana was obtained from the TAIR website, all BTB protein sequences of Paulownia and Arabidopsis thaliana were compared using a multiple sequence comparison tool, and a phylogenetic tree was constructed using MEGA-X;

[0011] S3. Gene structure and conserved motif analysis: Gene structure data were obtained from the Paulownia genome database to identify the introns, exons, and UTR structures of the PfBTB gene family. TBtools was used to visualize the gene structure and conserved motif analysis results.

[0012] S4. Localization of PfBTB gene on chromosome and analysis of homologous genes: The full-blast method and full-blast software were used to find homologous gene pairs, MCScanX was used to determine synchronous regions, and TBtools was used to analyze the chromosome position to show the distribution of synchronous gene pairs;

[0013] S5. Study on the expression pattern of BTB gene of Paulownia tomentosa: The transcription amount of BTB gene in Paulownia tomentosa seedlings with clustering disease and healthy Paulownia tomentosa seedlings was detected to obtain the transcription results of each gene.

[0014] The beneficial effects of the present invention are as follows:

[0015] The paulownia BTB gene family of the present invention and its application in biological and abiotic stresses can reduce the disease resistance rate of paulownia by obtaining genes related to resistance to paulownia witches' broom, and play an important guiding role in the subsequent cultivation of disease-resistant paulownia varieties. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 The chromosomal location of the Paulownia BTB gene family.

[0017] Figure 2 This is the phylogenetic tree of the BTB family of Paulownia alba and Arabidopsis thaliana.

[0018] Figure 3 Figure 2 shows the detection of conserved motifs in the BTB gene (left) and analysis of the UTR, intron and exon regions of the BTB gene (right).

[0019] Figure 4 This is the phylogenetic tree of the Paulownia BTB gene.

[0020] Figure 5 This is a synchronous study diagram of the distribution pattern of the BTB gene family.

[0021] Figure 6 It is a bar graph of the relative expression levels of Paulownia BTB genes in the stems, roots, and leaves of PF and PFI. DETAILED DESCRIPTION

[0022] Embodiments of the present invention are described in detail below, and examples of the embodiments are shown in the accompanying drawings. The embodiments described below with reference to the accompanying drawings are exemplary and intended to be used to explain the present invention, but should not be understood as limiting the present invention.

[0023] The present invention provides a Paulownia BTB gene family, which includes PfBT1-PfBT62. The Paulownia BTB gene has an important defense effect on biological stress. Figure 1 The Paulownia BTB gene is unevenly distributed on the 17 chromosomes, with the most distribution on chromosome 8 and the least distribution on chromosomes 18 and 19, each of which contains only one BTB gene. Chromosomes 1 and 20 do not contain any BTB genes, so there is no BTB gene in the chromosomes. Figure 1 Display them in .

[0024] A Paulownia BTB protein containing a Paulownia BTB gene, wherein the Paulownia BTB protein comprises a BTB domain and an ankyrin repeat sequence, and has both transcriptional activation and transcriptional inhibition effects. Figure 2 , Paulownia BTB proteins can be divided into 8 groups: MATH, Armadillo, BTB-only, TAZ, BACK, Ankyrin, TPR or NPH3, see Figure 3 , these proteins are highly conserved, UTRs are represented by green rectangles, exons are represented by yellow rectangles, and introns are represented by gray lines. The scale bar shows the value of 2kb.

[0025] The application of the Paulownia BTB gene family in biotic and abiotic stresses is to improve plant disease resistance by overexpressing the Paulownia BTB gene in target plants. Under diseased and healthy stresses, PfBTB12, PfBTB14 and PfBTB44 are significantly upregulated, and PfBTB3, PfBTB16, PfBTB19 and PfBTB36 are significantly downregulated.

[0026] A method for identifying a Paulownia BTB gene, comprising:

[0027] S1. Obtain BTB genes in Paulownia: Download the Paulownia genome from NCBI (https: / / www.ncbi.nlm.nih.gov / ), use HMM search software, use the seed file of BTB domain (PF00651) to search for the inferred BTB genes in the Paulownia genome database, upload all candidate proteins to the SMART website (http: / / smart.embl-heide lberg.de / ) to check the accuracy of the BTB conserved domain;

[0028] S2. Phylogenetic analysis of Paulownia BTB protein: The BTB protein sequence of Arabidopsis thaliana was obtained from the TAIR website (https: / / www.Arabidopsis.org / browse / genefamily / pub.jsp). All BTB protein sequences of Paulownia thaliana and Arabidopsis thaliana were compared using the multiple sequence alignment tool ClustalW. A phylogenetic tree was constructed using MEGA-X (Method, NJ; Bootstrap, 1000 was used, see ). Figure 4 , the evolutionary tree of BTB proteins was modified using the iTOL online website (https: / / itol.embl.de / );

[0029] S3. Gene structure and conserved motif analysis: Gene structure data were obtained from the Paulownia genome database to identify the introns, exons and UTR structures of the PfBTB gene family. TBtools was used to visualize the results of gene structure and conserved motif analysis. The position 2000 bp upstream of PfBTB was once described as a putative promoter sequence. Such PfBTB sequences were obtained using TBtools. The factors of the promoter positions of these genes were estimated using the promoter model. According to the characteristic annotations of cis-acting elements, cis-acting elements with the same functional annotations were built into the same group.

[0030] S4. Localization of PfBTB gene on chromosomes and analysis of homologous genes: Homologous gene pairs were discovered using the full-blast method and full-blast software, synchronous regions were determined using MCScanX, and chromosomal positions were analyzed using TBtools to show the distribution of synchronous gene pairs; see Figure 5 , The results of synchronous study on the distribution pattern of BTB gene family. The synchronous gene pairs of BTB gene family are represented by black lines.

[0031] S5. Study on the expression pattern of BTB gene of Paulownia tomentosa: The transcription amount of BTB gene in Paulownia tomentosa seedlings with clustering disease and healthy Paulownia tomentosa seedlings was detected to obtain the transcription results of each gene. The study showed that BTB gene plays an important role in defense against biological stress. Figure 6The relative expression levels of Paulownia BTB genes in stems, roots and leaves of PF and PFI are shown in the figure. The values ​​are expressed as the mean ± SE of three biological replicates. Different letters for each gene indicate significant differences (P < 0.05). PF represents healthy brown-spotted poplar seedlings. PFI represents Plasmodium fowleri seedlings infected with witches' broom phytoplasma.

[0032] Although the above embodiments have been shown and described, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations on the present invention. Changes, modifications, substitutions and variations of the above embodiments by those of ordinary skill in the art are all within the scope of protection of the present invention.

Claims

1. A Paulownia BTB gene family, characterized in that: The Paulownia BTB gene family includes PfBT1-PfBT62, and the Paulownia BTB gene has an important defense effect on biological stress.

2. The Paulownia BTB protein comprising the Paulownia BTB gene according to claim 1, characterized in that: The Paulownia BTB protein contains a BTB structural domain and an ankyrin repeat sequence, and has both transcription activation and transcription inhibition effects.

3. Application of Paulownia BTB gene family in biotic and abiotic stresses, characterized in that: By overexpressing the Paulownia BTB gene in the target plant, the disease resistance of the plant is improved. Under the stress of disease and health, PfBTB12, PfBTB14 and PfBTB44 are significantly upregulated, and PfBTB3, PfBTB16, PfBTB19 and PfBTB36 are significantly downregulated.

4. A method for identifying the BTB gene of Paulownia tomentosa, characterized in that: include: S1. Obtaining BTB genes in Paulownia: Download the Paulownia genome from NCBI, use HMM search software, and use the seed file of the BTB domain to search for the inferred BTB genes in the Paulownia genome database; S2. Phylogenetic analysis of Paulownia BTB protein: The BTB protein sequence of Arabidopsis thaliana was obtained from the TAIR website, all BTB protein sequences of Paulownia and Arabidopsis thaliana were compared using a multiple sequence comparison tool, and a phylogenetic tree was constructed using MEGA-X; S3. Gene structure and conserved motif analysis: Gene structure data were obtained from the Paulownia genome database to identify the introns, exons, and UTR structures of the PfBTB gene family. TBtools was used to visualize the gene structure and conserved motif analysis results. S4. Localization of PfBTB gene on chromosome and analysis of homologous genes: The full-blast method and full-blast software were used to find homologous gene pairs, MCScanX was used to determine synchronous regions, and TBtools was used to analyze the chromosome position to show the distribution of synchronous gene pairs; S5. Study on the expression pattern of BTB gene of Paulownia tomentosa: The transcription amount of BTB gene in Paulownia tomentosa seedlings with clustering disease and healthy Paulownia tomentosa seedlings was detected to obtain the transcription results of each gene.

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