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Transgenic method for improving cutting rooting rate of white birch

A technology of rooting rate and birch, applied in the directions of biochemical equipment and methods, genetic engineering, plant genetic improvement, etc., can solve the problems of high technical and conditional requirements, large-scale supply of planting raw materials, unsatisfactory rooting rate, etc. The effect of increasing the rooting rate

Active Publication Date: 2021-07-09
NORTHEAST FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The selection and breeding of fine varieties of birch began during the "Eighth Five-Year Plan" period. Remarkable results have been achieved in provenance selection and hybrid breeding. However, the promotion and utilization of improved varieties has been limited. The main reason is the lack of economical and effective asexual reproduction methods.
The birch asexual reproduction technology system has not been established, the tissue culture system has been established, and the somatic embryogenesis technology system is still in the exploratory stage, but the technical and conditional requirements for these two methods are high, which is not conducive to the promotion and application of grassroots units
The cutting propagation technology suitable for grassroots units, because the problem of birch cutting rooting has been difficult to break through
This method requires hormone treatment, and the achieved rooting rate is far from meeting the supply of large-scale planting raw materials. Therefore, the problem of cutting rooting is still not fundamentally solved, which restricts the promotion process of improved varieties of birch

Method used

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  • Transgenic method for improving cutting rooting rate of white birch
  • Transgenic method for improving cutting rooting rate of white birch
  • Transgenic method for improving cutting rooting rate of white birch

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Construction of BpmiR156 Overexpression Birch Strains

[0034] 1. Cloning of BpmiR156 gene

[0035] According to birch genome and transcriptome data (https: / / genomevolution.org / CoGe / GenomeInfo.pl?gid=35080), gene-specific primers miR156-F: 5'-CACCGTTTCCACATCTGGGAATAAGC-3' and miR156-R: 5'-TCAGCAGCTGTTGAAGTTGAAGG were designed -3'.

[0036] The birch cDNA was used as a template to amplify a 251bp fragment containing the mature sequence of BpmiR156, and the PCR band was detected by 1% agarose gel electrophoresis to obtain the target fragment.

[0037] PCR reaction system: 2μL 10×KOD Buffer, 2μL dNTPs (2.5mmol / L), 1.2μL MgSO 4 , 0.3 μL KOD Plus Neo, 0.6 μL miR156-F (10 μmol / L), 0.6 μL miR156-R (10 μmol / L), 2 μL template and 11.3 μL water.

[0038] PCR amplification reaction program: pre-denaturation at 94°C for 2min, denaturation at 94°C for 20s, annealing at 58°C for 20s, extension at 72°C for 15s, and extension at 72°C for 7min after 35 cycles.

[0039] The...

Embodiment 2

[0049] Example 2 Measurement of different physiological indexes of BpmiR156 birch

[0050] 1. Experimental materials

[0051] 1. Rooting and anatomical identification of BpmiR156 birch cuttings

[0052] Take the transgenic birch line with overexpression of miR156 and the non-transgenic control white birch young shoots cut into cuttings containing 2 bud points, in the substrate treated with carbendazim (50%) (black soil: peat soil: vermiculite = 2: 2:1) medium cuttings, attached with plastic wrap, placed in a plant culture room for cultivation, the cultivation conditions are: room temperature 25°C, 16h light / 8h dark. After 30 days of cutting, the rooting rate of the miR156-overexpressed birch line and control birch cuttings was counted. And at 0d, 3d, 6d, 9d, 12d, 15d, 18d, 21d after cutting, cut the 5mm long stem segments at the base of the cuttings for the preparation of paraffin sections

[0053] 2. Experimental methods and results

[0054] 1. Distribution of endogenous ...

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Abstract

The invention discloses a transgenic method for improving the cutting rooting rate of white birch, and belongs to the technical field of forest tree breeding. The invention provides the application of a white birch BpmiR156 gene in white birch cutting propagation breeding. A nucleotide sequence of the white birch BpmiR156 gene is shown as SEQ ID NO: 1. According to the invention, the BpmiR156 gene is introduced for overexpressing the white birch, so that the rooting rate can reach 85% or more under the condition that hormone treatment is not needed; the method is applied to white birch cutting propagation breeding, the white birch cutting propagation rooting rate can be remarkably increased, the bottleneck that white birch cutting rooting is difficult is effectively solved, and technical support is provided for white birch improved variety popularization.

Description

technical field [0001] The invention relates to the field of tree breeding, and relates to a transgenic method for increasing the rooting rate of white birch cuttings. Background technique [0002] At present, developed countries such as the United Kingdom, New Zealand, Canada and Brazil all put the utilization of heterosis in a priority position when developing industrial timber forests. After the 1980s, with the development of asexual reproduction technology, the United Kingdom, the United States, New Zealand, Canada and other countries have carried out research on the cutting propagation of some difficult-to-root tree species, especially in the application of broad-leaved tree hybrids to increase timber production and realize afforestation varieties. Achievements that have attracted worldwide attention. A number of high-yield and high-quality examples have been created in the aspect of "sexual seed production and asexual utilization". [0003] White birch (Betula platyp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/84C12N15/29A01H5/06A01H6/00
CPCC07K14/415C12N15/8205
Inventor 李慧玉李月马庆姜静刘桂丰
Owner NORTHEAST FORESTRY UNIVERSITY
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