Use of a ccch domain to increase biomass of poplar trees

Abstract

The application relates to the fields of molecular biology and genetic engineering technology, and particularly relates to a DNA sequence with a regulation function identified from a poplar 'Nanlin 895' variety, and research on the function and application of the DNA sequence. The cDNA sequence is shown as SEQ ID NO. 1. The application also discloses a method for promoting the growth of the poplar and regulating the yield of the poplar by using the segment.

Description

Technical Field

[0001] This invention belongs to the field of genetic engineering technology, specifically involving the transformation of a poplar tree... PdC3H17 Application of gene overexpression vectors containing CCCH domain fragment sequences in improving plant yield. Background Technology

[0002] Poplar is one of the most important plantation tree species in the world's temperate regions, possessing advantages such as rapid growth, strong adaptability, early maturity, and large timber volume. Furthermore, poplar has a rich genetic background, is easily propagated asexually, and undergoes genetic transformation. With the completion of whole-genome sequencing of Populus euphratica and Populus euphratica, poplar has become an ideal woody model plant. Therefore, researching methods to increase poplar yield is of great significance for improving timber production.

[0003] CCCH-type zinc finger proteins are a class of zinc finger proteins widely distributed in organisms, found in plants, animals, and microorganisms. These proteins all contain one or more conserved domains consisting of three cysteine ​​residues (Cys) and one histidine residue (His). In plants, they exist as multi-gene families and participate in the regulation of plant growth, development, and abiotic stress. For example, in Arabidopsis thaliana, SOMNUS has been shown to be a component of the phytochrome signaling pathway, negatively regulating light-dependent seed germination; PEI1 is involved in the formation of heart-shaped embryos; HUA1 plays an important regulatory role in flower development; AtSZF1 and AtSZF2 are involved in regulating Arabidopsis thaliana's tolerance to salt stress. AtC3H14 and AtC3H15 redundantly regulate stem secondary cell wall formation and anther development. In rice, OsLIC regulates plant morphogenesis; OsDOS affects leaf senescence through the jasmonic acid signaling pathway; and OsC3H12 positively regulates rice's quantitative resistance to Xoo.

[0004] Our previous systematic analysis of the poplar CCCH gene family revealed that poplar contains 91 members, evolutionarily divided into 13 subfamilies, including 6 members highly expressed in the xylem. Among them, Pd C3H17 contains two tandem CX8CX5CX3CH domains and is a typical CCCH zinc finger protein. Evolutionarily, it is homologous to Arabidopsis thaliana AtC3H14 and AtC3H15. Pd The C3H17 protein can bind to both DNA and RNA, and is primarily located in the cytoplasmic foci. It is specifically expressed in the xylem; overexpression promotes secondary xylem development and increases secondary cell wall thickness, while dominant inhibition produces the opposite phenotype, with the stem radius reduced by approximately one-fifth compared to the control. It is worth noting that... Pd C3H17 is a second-level regulatory hub in wood formation. Pd MYB3 andPd MYB21's direct target. Pd While C3H17 overexpression promotes stem thickening, it also leads to plant dwarfing, resulting in a slight decrease in timber yield. We analyzed... Pd After characterizing the C3H17 protein sequence, a vector was constructed to overexpress the CCCH domain, thereby creating a high-yield transgenic poplar tree. This is the main content of this patent. Summary of the Invention

[0005] The purpose of this invention is to provide a method for increasing poplar yield.

[0006] To achieve the above objectives, the present invention adopts the following technical solution: In a first aspect, the present invention provides a poplar tree PdC3H17 The CCCH domain fragment of the gene, the sequence of which is shown in SEQ ID NO.1.

[0007] In a second aspect, the present invention provides a material comprising poplar PdC3H17 Gene overexpression vectors for CCCH domain fragments, including microbial expression vectors and plant expression vectors.

[0008] In a third aspect, the present invention provides a poplar tree PdC3H17 Application of the CCCH domain fragment in increasing poplar yield.

[0009] This invention identified a stem xylem-specific expression in poplar. PdC3H17 The gene was selected, and an overexpression vector was constructed using the CCCH domain to promote poplar growth. Therefore, this invention helps to increase forest biomass. Attached Figure Description

[0010] The illustrative drawings that constitute a part of this invention are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an improper limitation of the invention.

[0011] Figure 1 For overexpression PdC3H17 Improving poplar timber yield after passing through the CCCH domain fragment of the gene Detailed Implementation

[0012] It should be noted that the following detailed descriptions are exemplary and intended to provide further illustration of the invention. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0013] It should be noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of exemplary embodiments according to the invention. As used herein, the singular form is intended to include the plural form as well, unless the context clearly indicates otherwise. Furthermore, it should be understood that when the terms "comprising" and / or "including" are used in this specification, they indicate the presence of features, steps, operations, components, and / or combinations thereof.

[0014] To promote the research and application of genetic engineering in increasing forest yield, the first aspect of this invention provides a poplar tree... PdC3H17 The CCCH domain fragment of the gene, the sequence of which is shown in SEQ ID NO.1.

[0015] In a preferred embodiment of the present invention, the poplar tree is amplified. PdC3H17 The CCCH domain fragment of the gene includes the following steps: (1) Poplar PdC3H17 Primers were designed using the CCCH domain fragment of the gene as a template. (2) Total RNA was extracted from poplar seedlings and then reverse transcribed into cDNA; (3) Using the cDNA obtained in step (2) above as a template, perform PCR amplification with a high-fidelity enzyme, recover and purify the product, and obtain the product.

[0016] In a second aspect, the present invention provides a material comprising poplar PdC3H17 Gene overexpression vectors for CCCH domain fragments, including microbial expression vectors and plant expression vectors.

[0017] In a third aspect, the present invention provides a poplar tree PdC3H17 Application of the CCCH domain fragment in increasing poplar yield.

[0018] To enable those skilled in the art to better understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below with reference to specific embodiments.

[0019] Example 1: Poplar PdC3H17 Cloning of the CCCH domain fragment of the gene and construction of an overexpression vector (1) Download poplar trees from the phytozome website PdC3H17 The cDNA sequence of the gene is analyzed, its specific regions are analyzed, and primers are designed to amplify them; (2) Total RNA was extracted from poplar seedlings using the Tiangen RNA Extraction Kit, and then the RNA was reverse transcribed into first-strand cDNA using the Total Gold Reverse Transcription Kit. (3) Using cDNA as a template, PCR amplification was performed using a high-fidelity enzyme, the purified product was recovered and constructed into an overexpression vector, and the sequencing results are shown in SEQ ID NO.1. sequence list <110> Qingdao Agricultural University <120> An application of CCCH domain to increase poplar biomass <141> 2021-08-21 <160> 1 <170> SIPOSequenceListing 1.0 <210> 1 <211> 237 <212> DNA <213> Poplar (2 Ambystoma laterale x Ambystoma jeffersonianum) <400> 1 gggatgtgga agacagagct atgcaacaag tggcaagaga caggcacctg tccttatggt 60 aaccattgtc agtttgctca tggcattggc gagctacgtc cagttatcag gcacccaagg 120 tacaagactc aagcttgccg catggttctt gctggcgggg tttgccctta tggccacaga 180 tgccactttc gccactccct cactgaccaa gagaggttac tgctgggtcc tcgttga 237

Claims

1. The application of a poplar PdC3H17 protein CCCH domain overexpression vector, characterized in that... The vector was genetically transformed into poplar trees, and the nucleotide sequence of the CCCH domain is shown in SEQ ID NO.1.

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