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Method capable of improving masson pine somatic embryo inductivity

A masson pine and induction rate technology, applied in the field of plant reproduction, can solve the problems of low induction rate of embryogenic callus, difficult to obtain regenerated plants, low germination rate, etc. high rate effect

Active Publication Date: 2018-10-19
GUANGXI FORESTRY RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there have been related studies on somatic embryogenesis of Pinus massoniana, but there are still problems such as low induction rate of embryogenic callus, difficulty in differentiation, low germination rate, and difficulty in obtaining regenerated plants.

Method used

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  • Method capable of improving masson pine somatic embryo inductivity
  • Method capable of improving masson pine somatic embryo inductivity

Examples

Experimental program
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Effect test

Embodiment 1

[0025]On June 25, 2015, the immature cones of Tongmian pine excellent tree that were healthy, free from diseases and insect pests, and the zygotic embryo was in the stage of fission and multiple embryos were collected as explants in the excellent stand of Tongmian pine in the state-owned Paiyangshan Forest Farm in Guangxi. Use volume concentration 45% alcohol + volume concentration 0.03% streptomycin + 100 mg·L -1 After the nicotinic acid mixture was wiped and disinfected on the surface of the explants, put every 4-5 explants into a plastic ziplock bag with a size of 20 mm×30 mm, seal the bag mouth by 80%, fold the bag mouth down Store in a refrigerator at 8°C for 14 days.

[0026] The pretreated explants were taken out of the refrigerator and rinsed with running water for 2 h, soaked in a broad-spectrum fungicide with a volume concentration of 0.1-0.3 % for 1 h, took out the explants and placed them in a volume concentration of 60 % alcohol for 3-5 min, then transferred to 8...

Embodiment 2

[0030] On July 15, 2016, the immature cones of the Tongmian pine excellent tree that were healthy, free from diseases and insect pests, and the zygotic embryos were in the stage of fission and multiple embryos were collected as explants in the excellent stand of Tongmian pine in the state-owned Paiyangshan Forest Farm in Guangxi. Use volume concentration 45% alcohol + volume concentration 0.03% streptomycin + 100 mg·L -1 After the nicotinic acid mixture was wiped and disinfected on the surface of the explants, put every 4-5 explants into a plastic ziplock bag with a size of 20 mm×30 mm, seal the bag mouth by 80%, fold the bag mouth down Store in a refrigerator at 6°C for 12 days.

[0031] The pretreated explants were taken out from the refrigerator and rinsed with running water for 2 h, soaked in a broad-spectrum fungicide with a volume concentration of 0.1-0.3 % for 1.5 h, took out the explants and placed them in a volume concentration of 60 % alcohol for 3-5 min, then trans...

Embodiment 3

[0035] On July 8, 2017, the immature cones of Tongmian pine excellent tree that were healthy, free from diseases and insect pests, and the zygotic embryo was in the stage of fission and polyembryony were collected as explants in the excellent stand of Tongmian pine in the state-owned Paiyangshan Forest Farm in Guangxi. Use volume concentration 45% alcohol + volume concentration 0.03% streptomycin + 100 mg·L -1 After the nicotinic acid mixture was wiped and disinfected on the surface of the explants, put every 4-5 explants into a plastic ziplock bag with a size of 20 mm×30 mm, seal the bag mouth by 80%, fold the bag mouth down Store in a refrigerator at 7°C for 14 days.

[0036]The pretreated explants were taken out of the refrigerator and rinsed with running water for 2 h, soaked in a broad-spectrum fungicide with a volume concentration of 0.1-0.3 % for 1 h, took out the explants and placed them in a volume concentration of 60 % alcohol for 3-5 min, then transferred to a sodi...

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Abstract

The invention provides a method capable of improving masson pine somatic embryo inductivity. The method comprises the work procedures: pretreating explant, sterilizing the expalnt and inducing a somatic embryo; collecting masson pine superior tree immaturate cones as the explant, pretreating and sterilizing the explant, peeling female gametophyte in the explant cones in a sterile mode, flatly putting the female gametophyte in an induction medium and putting into a suitable environment to be cultured, wherein the immaturate cones are healthy and have no pest and disease damage, zygotic embryo of the immaturate cones are in a cleavage polyembryony period, and an embryogenic callus induction culture period is 3 to 4 weeks; finally, obtaining a masson pine embryonic cell line which has quickness in growth and exuberant vigor. According to the method disclosed by the invention, a series of optimized explants are treated and sterilized, then female gametophyte in the cones is peeled and induced to form a lot of embryonic cell lines with exuberant vigor, powerful material resources and technical supports are established for further developing masson pine somatic embryo seedling culture, germplasm innovation and bioengineering seed breeding, and better economic benefits, social benefits and ecological benefits are achieved.

Description

technical field [0001] The invention belongs to the technical field of plant propagation and relates to tissue culture somatic embryogenesis technology, in particular to a method for increasing the induction rate of masson pine somatic embryos. Background technique [0002] Masson pine ( Pinus massoniana Lamb.) is the main tree species used for ecological construction and afforestation in southern my country. As a tree species with high comprehensive utilization value and broad promotion prospects, masson pine can not only be used to produce three-board, papermaking and chemical fiber industrial manufacturing, but also can be used to produce industrial raw materials such as rosin and turpentine. As a large forestry province, Guangxi, according to the "13th Five-Year" forestry development plan of the autonomous region, the development of 100 billion yuan of characteristic and advantageous industries such as turpentine industry and timber industry occupies a pivotal position...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/005
Inventor 姚瑞玲王胤
Owner GUANGXI FORESTRY RES INST
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