Tissue culture multiplication method for Pinus densiflora

A technology of tissue culture and red pine, applied in horticultural methods, botany equipment and methods, horticulture, etc., can solve the problems of low proliferation coefficient of red pine tissue culture, precious mature seeds of red pine, and difficulty in large-scale multiplication, and achieve high reproduction rate industrialization The effects of production, shortening of the culture cycle, and stable genetic traits

Inactive Publication Date: 2009-07-22
NANJING FORESTRY UNIV
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  • Application Information

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Problems solved by technology

[0006] Purpose of the invention: the present invention mainly aims at the problems that mature seeds of red pine resistant to pine wood nematode are precious and rare, and the proliferation coefficient of r

Method used

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  • Tissue culture multiplication method for Pinus densiflora

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Selection of Akapine 1 imported from Japan that is resistant to pine wood nematode disease # 、7 # The mature seeds of Youshu were treated at a low temperature of 4°C for 4 weeks, taken out and pretreated with 0.1% potassium permanganate for 12 hours, washed with running water for 1 hour, after the washing was completed, the seed shells were peeled off under sterile conditions, and the surface was treated with 70% ethanol After sterilizing for 30 seconds, soak in 0.1% mercuric chloride for 2 minutes, rinse with sterile water for 4 to 5 times, and transfer to culture medium to grow aseptic vaccines for later use.

[0020] Select aseptic seedlings that have germinated for 3 to 4 weeks, cut off the hypocotyl at 6 to 8 mm below the cotyledon, and use the cotyledon node (containing the cotyledon and just germinated epicotyl and 6 to 8 mm hypocotyl) as an explant, Inoculate in GD+6-BA3.0~5.0mg / L+NAA0.1~0.3mg / L medium, after 4~5 weeks, transfer the explants together with the d...

Embodiment 2

[0025] Selection of Akapine 5 imported from Japan that is resistant to pine wood nematode disease # 、16 # For the mature seeds of excellent trees, on the basis of Example 1, the medium that promotes the elongation of clustered buds is replaced with DCR+NAA0.1~0.3mg / L, and the rest of the steps are the same as in Example 1, and the results are equivalent to those in Example 1. Effect.

[0026] The following specific experimental situations will help to illustrate the effectiveness of the formula, explant cutting mode and subculture mode used in the method of scorching pine tissue culture and proliferation of the present invention, and all tests are repeated at least 3 times.

[0027] First, the effect of hormones on the proliferation of clustered buds of Pinus chinensis

[0028] Take GD as the basic medium, add 6-BA0.5~2.5mg / L+NAA0.25mg / L or kinetin (KT)0.5~2.5mg / L+NAA0.25mg / L, and compare the effects of different hormone combinations on clustered buds. effects on proliferat...

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Abstract

The invention relates to a red pine tissue propagation method. The method comprises the following steps: taking a red pine asepsis seedling cotyledonary node (containing a cotyledon, a budding epicotyl and a 6 to 8mm hypocotyl) as an explant which is inoculated in a GD+6-BA3.0 to 5.0mg/L+NAA0.1 to 0.3mg/L culture medium so as to induce a cotyledon base to generate clustered seedlings; after 4 to 5 weeks, shifting the explant together with the differentiated clustered seedlings into a culture medium with added active carbon or NAA so as to promote the elongation of the seedlings; and when the clustered seedlings grow to 1.5 to 2.0cm, cutting off the seedlings separately, cutting the seedlings into 5 to 8mm small sections, and inoculating the small sections in a GD+6-BA1.5 to 2.5mg/L+NAA0.25mg/L culture medium for further inducing the propagation of clustered seedlings. The propagated clustered seedlings can be used for a new cycle of propagation again after culture and elongation. The method can realize the clustered seedlings propagation coefficient of between 3.5 and 4.6; and a seed can propagate 20 to 30 thousand tissue culture seedlings for rootage 2 years later.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically refers to a method for culturing and proliferating red pine tissue. Background technique [0002] Pinus (Pinus) is the largest and most important genus among the genus of conifers. There are about 116 species, varieties and hybrids. It is widely distributed and planted in the world. , and has a very important position in the ecosystem. Due to the harm of various diseases and insect pests, a large number of pine trees die every year, among which the pine wood nematode (Bursaphelenchus xylophilus) is the most serious harm. The disease has now caused a total area of ​​2.66 million ha of Japanese pine forests 2 450,000 hm in 2 morbidity, the loss is very heavy. In my country, the total number of dead pine trees caused by pine wood nematode disease has reached more than 50 million, with a direct economic loss of 2.5 billion yuan and an indirect economic loss of 25 billion yuan, and it...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 吴小芹朱丽华叶建仁戴培培
Owner NANJING FORESTRY UNIV
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