Tissue culture and regenerated plant in vitro mycorrhization method for pinus massoniana

A technology for plant regeneration and tissue culture, applied in the fields of biotechnology and modern forestry, can solve the problems of lack of plant regeneration system and mycorrhization technology in bottles, low mycorrhization rate, and limited application, so as to increase the probability of contact, Effect of increasing mycorrhization rate and improving quality

Inactive Publication Date: 2009-11-25
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Cheng Xiaofei, Zheng Laiyou, etc. have carried out preliminary research on the mycorrhizal synthesis of Pine massoniana tissue culture regeneration plants (Cheng Xiaofei et al., 1995; Zheng Laiyou et al., 2003), but the matrix that it adopts is agar medium, and mycelium stays in mostly The surface of the medium is difficult to contact with adventiti

Method used

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  • Tissue culture and regenerated plant in vitro mycorrhization method for pinus massoniana
  • Tissue culture and regenerated plant in vitro mycorrhization method for pinus massoniana
  • Tissue culture and regenerated plant in vitro mycorrhization method for pinus massoniana

Examples

Experimental program
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Effect test

Embodiment 1

[0024] Take the mature seeds of Pinus massoniana, wash them with detergent, wash them with running water for 1 hour, peel off the seed husks under sterile conditions, sterilize the surface with 70% ethanol for 30-35 seconds, and then soak them in 0.1% mercury liter for 3-4 minutes. After rinsing with sterile water for 3 to 5 times, transfer to agar medium to germinate into sterile seedlings. Take the terminal buds of sterile seedlings that have germinated for 3 weeks, remove the top growth point, keep the cotyledons and 5mm hypocotyls, and inoculate them in GD+6-BA3mg / L (up to 4mg / L)+NAA0.1mg / L (up to 0.3 mg / L) + sucrose 3% medium to induce clustered buds. After 4 weeks of cultivation, transfer the terminal buds of the aseptic seedlings that have produced clustered buds into 1 / 2GD+AC0.5g / L (up to 1.0g / L)+2% sucrose medium, and cultivate for 4 weeks until the buds grow 1.0- 1.5cm. Individually cut out the shoots with a length of 1.0-1.5cm, and inoculate them into 1 / 4GD+NAA0.1...

Embodiment 2

[0028] Repeat the same steps as described in Example 1, but replace the adventitious root induction medium with 1 / 2SH+NAA0.2mg / L+IBA1.0mg / L+sucrose 1%, and replace the liquid medium added to the perlite matrix with 1 / 2 SH (containing 0.5% sucrose), can obtain the effect equivalent to that of Example 1.

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Abstract

The invention discloses a tissue culture and regenerated plant in vitro mycorrhization method for pinus massoniana, which belongs to the technical field of biotechnology and modern forestry. The method comprises the following steps of: taking a terminal bud of an aseptic seedling of the pinus massoniana as an explant, inducing to produce a large quantity of multiple shoots with strong growth first, further inducing the generation of an adventitious root, transferring a plant with the macroscopic adventitious root onto a perlite substrate under the condition of asepsis after the induction of the adventitious root is finished, and inoculating ectomycorrhizal rungi Pisolithustinctorius at the same time to realize the mycorrhization in a bottle. The mycorrhization improves the quality of a root system of the adventitious root of a tissue culture regenerated plant of the pinus massoniana, and ensures that the average lateral root number and the average root length are greatly improved. The mycorrhization also makes the state of the adventitious root greatly changed, and a fungal mantle obtained by closely arranging more than ten layers of mycelia is formed on the surface of the adventitious root. Through the method, the induction rate of the multiple shoots of the pinus massoniana reaches 95 percent, the proliferation rate reaches 430 percent, the rooting rate reaches 85 percent, and the transplantation survival rate is improved to more than 85 percent from 65 percent.

Description

1. Technical field [0001] The patent of the invention relates to a method for synthesizing mycorrhizae of pine tissue culture and regenerated plants under aseptic conditions, and belongs to the field of biotechnology and modern forestry technology. 2. Background technology [0002] Masson pine (Pinus massoniana Lamb.) is an important timber tree species and resin-producing tree species in my country. It is also a pioneer tree species for afforestation on barren hills. It is distributed in 16 provinces (autonomous regions and municipalities) across the country, and its plantation area accounts for about 100% of the country's plantation area. It has a very important position in my country's forestry production. Masson pine mainly reproduces by seeds. Due to the low seed yield in the seed orchard and the large and small years, the supply of masson pine varieties is insufficient, and there are large differences in the growth of different geographical provenances. Its vegetative ...

Claims

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Application Information

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IPC IPC(8): A01H4/00A01G7/06A01G31/00C12N5/04
Inventor 吴小芹朱丽华叶建仁唐颖
Owner NANJING FORESTRY UNIV
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