Search

CN-122012533-A - Pt7G25060 gene and application of protein thereof in judging drought stress degree of Chinese pine seedlings

CN122012533ACN 122012533 ACN122012533 ACN 122012533ACN-122012533-A

Abstract

The invention discloses an application of Pt7G25060 gene and a protein thereof in determining drought stress degree of Chinese pine seedlings, and belongs to the technical field of biology. The gene expression quantity is inversely related to the drought stress degree, and the gene expression quantity shows obvious expression difference in the early drought stress stage of the Chinese pine seedlings. Based on the technical advantages, the invention provides a method for judging the drought stress degree of the Chinese pine seedlings. By using the detection primer provided by the invention to carry out fluorescent quantitative PCR amplification on the cDNA of the sample to be detected and the cDNA of the reference sample and comparing the expression quantity of the Pt7G25060 gene in the sample to be detected and the reference sample, the accurate judgment of the drought stress degree of the sample to be detected can be realized. Compared with the traditional resolution method which depends on the degree of observing seedling needles She Weihuang and physiological indexes, the method provided by the invention can sensitively and accurately reflect the drought stress degree of the pinus koraiensis seedlings, and has the advantages of higher precision, stronger scientificity and good repeatability.

Inventors

  • CUI YANING
  • ZHANG CE
  • LI SIBO
  • LIU XUANDONG
  • LU DUXIAN
  • XU CHANGWEN
  • LI YANJUN
  • LIN JINXING
  • ZHANG YANYAN
  • WANG XIAO

Assignees

  • 北京林业大学

Dates

Publication Date
20260512
Application Date
20260415

Claims (10)

  1. 1. The Pt7G25060 gene is characterized in that the amino acid sequence coded by the Pt7G25060 gene is shown as SEQ ID NO. 2.
  2. 2. The Pt7G25060 gene of claim 1, wherein the nucleotide sequence of the Pt7G25060 gene is set forth in SEQ ID No. 1.
  3. 3. The Pt7G25060 protein is characterized in that the amino acid sequence of the Pt7G25060 protein is shown as SEQ ID NO. 2.
  4. 4. The detection primer for the Pt7G25060 gene according to claim 1, wherein the nucleotide sequences of the detection primers are shown in SEQ ID NO.3 and SEQ ID NO.4 respectively.
  5. 5. A kit for amplifying the Pt7G25060 gene of claim 1, comprising the detection primer of claim 4.
  6. 6. The kit of claim 5, further comprising a PCR amplification reagent comprising SYBR Green DNA polymerase, dNTPs, PCR buffer, and internal reference gene primers.
  7. 7. The kit according to claim 6, wherein the nucleotide sequences of the reference gene primers are shown in SEQ ID NO.5 and SEQ ID NO.6, respectively.
  8. 8. Use of the Pt7G25060 gene of claim 1 or 2, the Pt7G25060 protein of claim 3, the detection primer of claim 4 or the kit of any one of claims 5 to 7 for determining the degree of drought stress to pine seedlings.
  9. 9. A method for determining the degree of drought stress to which a young pine seedling is subjected, comprising the steps of: respectively carrying out fluorescent quantitative PCR amplification by using the detection primers of claim 4 by taking cDNA of a sample to be detected and cDNA of a reference sample as templates, and respectively counting the expression amounts of Pt7G25060 genes in the sample to be detected and the reference sample; When the expression level of the Pt7G25060 gene in the sample to be tested is not significantly different from the expression level of the Pt7G25060 gene in the reference sample, judging that the drought stress degree of the sample to be tested is the same as the stress degree of the reference sample; when the expression quantity of the Pt7G25060 gene in the sample to be tested is obviously lower than that of the Pt7G25060 gene in the reference sample, judging that the drought stress degree of the sample to be tested is higher than that of the reference sample; When the expression quantity of the Pt7G25060 gene in the sample to be tested is obviously higher than that of the Pt7G25060 gene in the reference sample, judging that the drought stress degree of the sample to be tested is lower than that of the reference sample; the amino acid sequence of the Pt7G25060 gene is shown as SEQ ID NO. 2.
  10. 10. The method for determining the drought stress level of pine seedlings according to claim 9, wherein said reference samples are samples obtained by subjecting pine seeds to the same drought stress level and different drought stress times.

Description

Pt7G25060 gene and application of protein thereof in judging drought stress degree of Chinese pine seedlings Technical Field The invention belongs to the technical field of biology, and particularly relates to an application of a Pt7G25060 gene and a protein thereof in determining drought stress degree of Chinese pine seedlings. Background The pine (Pinus tabuliformis Carriere) belongs to pine of Pinaceae, needle She Changlu arbor, and has strong cold resistance, drought resistance and barren resistance. The Chinese pine has remarkable water and soil conservation and water conservation effects, can effectively prevent soil erosion, improve soil structure and enhance soil fertility, and also plays a key role in wind prevention and sand fixation, climate adjustment, vegetation restoration and the like, so that the Chinese pine becomes an important tree species for barren mountain forestation, water and soil conservation and urban greening. In addition, the Chinese pine timber has hard material, corrosion resistance and beautiful texture, and is widely used in the fields of building, bridge, railway sleeper, ore pillar, furniture manufacturing and the like. The pine resin, pine needle, pine knot, bark and other parts of the Chinese pine contain rich medicinal components, are commonly used for extracting turpentine, rosin and the like, have the effects of dispelling wind and removing dampness, activating blood and relieving pain, resisting bacteria and diminishing inflammation and the like, and have higher application value in Chinese medicinal materials and chemical products. However, as the climate is warmed up and aggravated, drought frequently occurs in many areas, and the normal running of the seedling raising of the Chinese pine is seriously affected. The economic loss of the drought stress to the Chinese pine artificial forest is not neglected, in the seedling raising and forestation process in arid and semiarid regions, the seedling growth is slow and the survival rate is reduced due to drought, and the seedling raising cost and the planting cost are directly increased. In addition, seedling death and stand decay caused by drought further aggravate the soil erosion risk, weaken the key effect of pine in soil and water conservation and ecological restoration, bring huge economic loss to forestry production and ecological environment construction. The Chinese pine has certain drought resistance, so that it is sometimes difficult to judge whether the Chinese pine is subjected to drought stress or not from the appearance phenotype in time. The cell wall is the first contact interface between the plant cells and the external environment and is therefore also the ideal location for sensing environmental changes, including water depletion. When drought occurs, changes in cell wall tension, pressure or structure are often caused, thereby activating receptor kinases on the cell membrane, initiating intracellular signal transduction. There is no clear decision at present whether there is a responsive gene marker for seedling drought stress in the cell wall of Pinus koraiensis. Therefore, the exploration of drought stress response markers in the seedling stage of the Chinese pine is a technical problem to be solved urgently by the person skilled in the art. Disclosure of Invention The invention discloses an application of Pt7G25060 gene and protein thereof in determining drought stress degree of Chinese pine seedlings. The method for judging the drought stress degree of the Chinese pine seedlings is an earlier and more accurate drought diagnosis method, and has important theoretical and practical significance for guiding scientific water management of Chinese pine seedlings, improving the survival rate of the seedlings, reducing the economic loss of forestry and guaranteeing the effect of ecological restoration engineering. In order to achieve the above object, the present invention provides the following technical solutions: a Pt7G25060 gene, wherein the amino acid sequence coded by the Pt7G25060 gene is shown as SEQ ID NO. 2. As a preferable technical scheme, the nucleotide sequence of the Pt7G25060 gene is shown as SEQ ID NO. 1. The invention also provides a Pt7G25060 protein, and the amino acid sequence of the Pt7G25060 protein is shown as SEQ ID NO. 2. The invention also provides a detection primer of the Pt7G25060 gene, and the nucleotide sequences of the detection primer are respectively shown as SEQ ID NO.3 and SEQ ID NO. 4. The invention also provides a kit for amplifying the Pt7G25060 gene, which comprises the detection primer. As a preferable technical scheme, the kit further comprises a PCR amplification reagent, wherein the PCR amplification reagent comprises SYBR Green DNA polymerase, dNTPs, a PCR buffer solution and an internal reference gene primer. As a more preferable technical scheme, the nucleotide sequences of the internal reference gene primers are respectively shown as SEQ ID NO.5 and SEQ ID