Tissue culture multiplication method for Pinus densiflora

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+1.5 to 2.5mg / L of 6-BA+0.25mg / L of NAA 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...

AI Technical Summary

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 red pine tissue culture is low, and it is difficult to multiply in large quantities, and researches and invents a method for the proliferation of red pine tissue culture to meet the needs of social production