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Ex-vitro rooting method for tissue culture seedlings of pinus massoniana

A technology for rooting out of a bottle and tissue culture seedlings, applied in the field of plant reproduction, can solve problems such as difficulty in meeting the needs of afforestation, inability to maximize the gain of genetic improvement, restricting the process of breeding of masson pine fine varieties, etc. Good economic benefits and the effect of reducing production costs

Active Publication Date: 2013-12-25
GUANGXI FORESTRY RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, sexual reproduction has the problem of differentiation and cannot maximize the genetic improvement gain, while asexual reproduction can overcome the above-mentioned shortcomings of sexual reproduction
However, the cutting and grafting of the traditional asexual propagation technology of Masson pine are still difficult to meet the needs of afforestation, which restricts the process of masson pine breeding.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] 1. Experimental materials

[0025] The terminal buds of aseptic seedlings germinated from the mature cone seeds of Pine masson in the same year were used as explants, and the cones were collected from Ningming 3, a high-yielding and excellent Pine masson family in the mother forest of Pine masson in Ningming, Guangxi.

[0026] 2. Experimental method

[0027] (1) Establishment of sterile culture system

[0028] Inoculate sterile explants into modified MS medium + 6-BA 3.0 mg·L -1 +NAA 0.1 mg·L -1 + sucrose 30000 mg·L -1 + Agar 3500 mg·L -1 cultured on culture medium. After 26 days of culture, the average cluster bud induction rate reached 99.1%.

[0029] The basic composition of the improved MS medium is: KNO 3 1650 mg·L -1 ; NH 4 NO 3 600 mg·L -1 ;CaCl 2 2H 2 O 260 mg·L -1 ;MgSO 4 ·7H 2 O 370 mg·L -1 ; Ca(NO 3 ) 2 4H 2 O 400 mg·L -1 ;KH 2 PO 4 170 mg·L -1 ;MnSO 4 ·H 2 O 22.3 mg L -1 ; ZnSO 4 ·7H 2 O 8.6 mg L -1 ; CuSO 4 ·5H 2 O 0.025 mg·...

Embodiment 2

[0039] 1. Experimental materials

[0040] The terminal buds of aseptic seedlings germinated from mature cone seeds of Pine masson in the same year were used as explants, and the cones were collected from Ningming pine family Ningming No.

[0041] 2. Experimental method

[0042] (1) Establishment of sterile culture system

[0043] Inoculate sterile explants into modified MS+6-BA 4.0 mg·L -1 +NAA 0.1 mg·L -1 + sucrose 30000 mg·L -1 + Agar 3500 mg·L -1 cultured on culture medium. After 26 days of culture, the average cluster bud induction rate reached 98.4%.

[0044] The basic composition of the improved MS medium is the same as in Example 1.

[0045] (2) Subculture multiplication and expansion

[0046] The induced clustered budlets were transferred to the improved MS+NAA 0.3 mg·L -1 + sucrose 30000 mg·L -1 + Agar 3500 mg·L -1 on elongation medium to promote elongation of the culture. After elongation culture for 28 days, the height of cluster buds is 3-5 cm. Cut out...

Embodiment 3

[0054] 1. Experimental materials

[0055] The top buds of aseptic seedlings germinated from the mature cone seeds of Pine masson in the same year were used as explants, and the cones were collected from Ningming 7, a high-yielding and excellent Pine masson family in the mother forest of Pine masson in Ningming, Guangxi.

[0056] 2. Experimental method

[0057] (1) Establishment of sterile culture system

[0058] Inoculate sterile explants into modified MS+6-BA 4.0 mg·L -1 +NAA 0.1 mg·L -1 + sucrose 30000 mg·L -1 + Agar 3500 mg·L -1 cultured on culture medium. After 26 days of culture, the average cluster bud induction rate reached 99.0%.

[0059] The basic composition of the improved MS medium is the same as in Example 1.

[0060] (2) Subculture multiplication and expansion

[0061] Transfer the induced clustered buds to the improved MS+AC 4000 mg·L -1 + sucrose 30000 mg·L -1 + Agar 3500 mg·L -1on elongation medium to promote elongation of the culture. After elonga...

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Abstract

The invention discloses an ex-vitro rooting method for tissue culture seedlings of pinus massoniana. The ex-vitro rooting method comprises the following steps: taking terminal buds of aseptic seedlings with 0.5cm long hypocotyls as explants for inducing cluster buds in a culture medium I; carrying out elongation culture on the induced cluster buds in a culture medium II, individually cutting off the terminal buds, and inoculating the cut terminal buds into a culture medium III for propagation to obtain subculture buds I; repeatedly elongating the subculture buds I, individually cutting off the subculture buds I, dividing the cut subculture buds I into terminal buds and bud sections, continuously propagating in a subculture propagation culture medium III to obtain subculture buds II, and repeatedly elongating and propagating the subculture buds II to obtain new propagation subculture buds; and transplanting the individually cut 2.5-3.5cm high subculture buds into a seedling culture cup for ex-vitro rooting, and transplanting the ex-vitro subculture buds into a seedling nursery for culture to obtain the ex-vitro tissue culture seedlings. According to the method disclosed by the invention, the production program of test-tube seedlings is simplified, the production cost is reduced, the production efficiency is improved, the rooting treatment time is 25-30 days, the ex-vitro rooting rate reaches 92%-98%, the root system quality is higher, the transplanting survival rate reaches 90%-95%, and the method has better economic, social and ecologic benefits.

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 rooting out of a bottle of tissue culture seedlings of Pinus massoniana. Background technique [0002] Masson pine ( Pinus massonianan ) is native to my country and has a very wide distribution range in my country. It is one of the important tree species in the important barren hills greening, papermaking raw material forests, resin mining forests, and natural landscape forests in southern 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, 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 resources an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 姚瑞玲袁铁象吴幼媚蔡玲
Owner GUANGXI FORESTRY RES INST
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