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Culture method for proliferation and rejuvenation of subculture buds in tissue culture of masson pine

A culture method and subculture technology, applied in the field of plant reproduction, can solve the problems of low effective bud proliferation coefficient, poor subculture bud physiological activity, difficulty in subculture proliferation, etc., and achieve high subculture proliferation coefficient and shorten subculture. The effect of cycle, culture medium science

Active Publication Date: 2018-12-11
GUANGXI FORESTRY RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the easy physiological aging of Pine massoniana in the in vitro tissue culture process, the physiological activity of the subcultured buds is poor, the subculture proliferation is difficult, the effective bud proliferation coefficient is low, and vitrification and browning are obvious, which seriously limits the excellent quality of Pine massoniana. Development of Industrialization of Germplasm Asexual Seedling Breeding

Method used

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  • Culture method for proliferation and rejuvenation of subculture buds in tissue culture of masson pine
  • Culture method for proliferation and rejuvenation of subculture buds in tissue culture of masson pine
  • Culture method for proliferation and rejuvenation of subculture buds in tissue culture of masson pine

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Effect test

Embodiment 1

[0025] According to the Guangxi local standard of tissue culture and rapid propagation of Pinus massoniana, routine disinfection, inoculation and subculture were carried out, and the subcultures were subcultured 3 times / subculture cycle 40 days, with stretched needles, longer leaf nodes, and 3.5-4.5 cm high subcultures. Substitute buds were used as experimental materials, and the tissue-cultured shoots of Pinus massoniana were subcultured and rejuvenated.

[0026] Select a 500 mL colorless glass triangular flask as the subculture bottle, add 100 mL of solid medium Ⅰ to the subculture bottle, and after routine high-temperature and high-pressure disinfection, cut off the vigorously growing subculture buds with a height of 2.5-3.5 cm , transferred to a subculture bottle containing solid medium Ⅰ, at a temperature of 25±1°C, the light source is a red and blue LED light of 650 nm red light: 450 nm blue light=3:5, and the light intensity is 3000-3500 lx, After culturing for 7 days u...

Embodiment 2

[0030] According to the Guangxi local standard of tissue culture and rapid propagation of Pinus massoniana, routine disinfection, inoculation and subculture were carried out, and the subcultures were subcultured 5 times / 40 days, with stretched needles, longer leaf nodes, and 3.5-4.5 cm high subcultures. Substitute buds were used as experimental materials, and the tissue-cultured shoots of Pinus massoniana were subcultured and rejuvenated.

[0031] Select a 500 mL colorless glass triangular flask as the subculture bottle, add 100 mL of solid medium Ⅰ to the subculture bottle, and after conventional high-temperature and high-pressure disinfection, cut off the vigorously growing subculture buds with a height of 2.5-3.5 cm , transferred to a subculture bottle containing solid medium Ⅰ, at a temperature of 25±1°C, the light source is a red and blue LED light of 650 nm red light: 450 nm blue light=3:5, and the light intensity is 3000-3500 lx, After culturing for 8 days under the con...

Embodiment 3

[0035] According to the Guangxi local standard of tissue culture and rapid propagation of Pinus massoniana, routine disinfection, inoculation and subculture were carried out, with subculture 10 times / subculture cycle 40 days, the needles stretched, the leaf nodes were longer, and the subcultures with a height of 3.5-4.5 cm were used. Substitute buds were used as experimental materials, and the tissue-cultured shoots of Pinus massoniana were subcultured and rejuvenated.

[0036] Select a 500 mL colorless glass triangular flask as the subculture bottle, add 100 mL of solid medium Ⅰ to the subculture bottle, and after routine high-temperature and high-pressure disinfection, cut off the vigorously growing subculture buds with a height of 2.5-3.5 cm , transferred to a subculture bottle containing solid medium Ⅰ, at a temperature of 25±1°C, the light source is a red and blue LED light of 650 nm red light: 450 nm blue light=3:5, and the light intensity is 3000-3500 lx, After culturin...

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Abstract

The invention provides a culture method for proliferation and rejuvenation of subculture buds in tissue culture of masson pine. The culture method comprises the following steps of selection of subculture buds, induction of axillary buds, and rejuvenation culture; selecting the subculture single bud of the masson pine after conventional disinfection, inoculation and subculture, transferring into asubculture bottle of a solid culture medium I, and performing induction of the axillary buds; adding a liquid culture medium II into the original subculture bottle by a double-layer and double-phase culture method, so as to obtain the subculture clumpy bud of the masson pine with strong growth vigor. The culture method has the advantages that the transferring step in the traditional subculture method of tissue culture of masson pine is simplified, and the subculture clumpy bud of the masson pine with high subculture proliferation coefficient, strong bud and seedling vigor and difficulty in aging is obtained; the quick propagation and utilization of scale in-vitro tissue culture of high-quality variety of masson pine are realized; the better economic, social and ecological benefits are realized.

Description

technical field [0001] The invention belongs to the technical field of plant propagation, and relates to tissue culture seedling raising technology, in particular to a method for the proliferation and rejuvenation of masson pine tissue culture subculture buds. Background technique [0002] Masson pine ( Pinus massonianan ) is native to my country and is one of the important tree species in barren hills greening, papermaking raw material forests, resin mining forests, and natural landscape forests in southern my country. It is widely distributed in my country. As a tree species with high comprehensive utilization value and great promotion potential, masson pine can not only be used to produce three-board, papermaking and chemical fiber industry manufacturing, but also can be used to produce industrial raw materials such as rosin and turpentine. In recent years, with the intensification of contradictions between people's growing living needs and the shortage of timber resourc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 姚瑞玲王胤
Owner GUANGXI FORESTRY RES INST
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