Method for somatic embryogenesis and plant regeneration of slash pines

A technique for embryogenesis and slash pine, applied in the field of plant tissue culture, can solve the problems of hindering the industrial application of the technology, low embryo induction rate, survival rate, somatic embryo maturation rate and germination rate, etc.

Active Publication Date: 2018-06-01
GUANGDONG ACAD OF FORESTRY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Somatic embryogenesis and plant regeneration of slash pine have been reported, but the embryo induction rate, survival rate, somatic embryo maturation rate and germination rate of slash pine in previous studies are generally low, which greatly hinders the industrial application of this technology

Method used

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  • Method for somatic embryogenesis and plant regeneration of slash pines
  • Method for somatic embryogenesis and plant regeneration of slash pines
  • Method for somatic embryogenesis and plant regeneration of slash pines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiment 1: Observation of the embryo maturation situation of slash pine cone seeds

[0049] Using 6 improved slash pine trees growing well, healthy and free from pests in the Hongling Seed Garden of Taishan City, Guangdong Province as material resources, of which 3 EE1 and 3 EE2 were harvested on June 16, June 23, 26, 2016, respectively. On July 30th and July 7th, collect cones, 3 to 9 cones per tree each time, store the cones collected on that day in an ice box at low temperature and bring them back to the laboratory, and then put them in a refrigerator at 4°C. Refrigerate in the refrigerator for observation of embryo maturity, the steps are as follows:

[0050] (1) Peel the cones and separate the explants containing zygotic embryos and endosperms: after the collected cones are cut in half, the reverse seed scales are pushed aside, the seeds are picked out, and the outer testa is peeled off to obtain the described Explant.

[0051] (2) Move the explants obtained i...

Embodiment 2

[0056] Example 2: Somatic embryogenesis and plant regeneration of immature zygotic embryos of slash pine

[0057]The formulations of the medium used in this example are shown in Table 2, wherein formulation 1 is an induction medium, formulation 2 is a proliferation medium, formulation 3 is a maturation medium, and formulation 4 is a germination medium.

[0058] Table 2 Components of each medium formula

[0059]

[0060]

[0061] Note: "—" in the table means that this component has not been added. After adding the formula components in the above Table 2, 0.5 g / L of Casein Hydrolysate and Phytagel are also added to Formula 1. Add 2g / L of Casein Hydrolysate to formulas 2, 3, and 4; add Phytagel 3g / L to formulas 3 and 4; adjust the pH of the medium to 5.70-5.80 with 1M HCl or NaOH.

[0062] (1) Cone collection

[0063] According to the optimal collection time of the cones determined in Example 1, a large number of cones were collected.

[0064] (2) Disinfection of peeled ...

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Abstract

The invention relates to a method for somatic embryogenesis and plant regeneration of slash pines. The method for somatic embryogenesis and plant regeneration of the slash pines comprises the steps of(1) collecting slash pines cones and isolating immature zygotic embryos; (2) inducing embryogenic callus formation; (3) proliferation of embryogenic callus; (4) curing and cultivating cotyledon embryos with germination potential; (5) sprouting and cultivating to obtain sprouted seedlings and (6) hardening seedlings and transplanting. An induction medium is a high-concentration nitrate nitrogen medium. The total concentration of nitrate ions in the culture medium is 13.8 to 14.6 mM; hormone components in the induction medium are prepared from 2.5 to 3 mu M of 6-benzyl aminopurine, 10.5 to 11 mu M of naphthylacetic acid, 2.5 to 3 mu M of Kinetin, 18.7 to 19.2 mu M of abscisic acid, and 0.8 to 1.2 mu M of brassinolide. The method has a high induction rate of embryogenic callus, somatic embryo maturation rate and germination rate, and the plant survival rate after transplanting is high.

Description

technical field [0001] The invention relates to the technical field of plant tissue culture, in particular to a method for pine cell embryogenesis and plant regeneration. Background technique [0002] Slash pine (Pinus elliottii Engelmann) is a tree species of the genus Pinus in the family Pinaceae. Arbor, native to the warm, humid and low-altitude areas of the southeastern United States, with a height of 30 meters and a diameter at breast height of 90 centimeters in the original place. Introduced to my country in the 1930s, it has the advantages of strong adaptability, drought resistance and barrenness, high afforestation survival rate, high rosin yield, good quality, and high economic value. After continuous improvement and promotion of planting, the current wetland Pine planting area reaches 2 million hectares 2 , distributed between 18°30' and 35°50' north latitude, is one of the main pine resin raw material cultivation tree species in southern my country. [0003] Imp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/005A01H4/008
Inventor 郭文冰赵奋成胡继文司徒荣贵彭冠明王为民黄婷邓乐平吴惠姗李义良廖仿炎
Owner GUANGDONG ACAD OF FORESTRY
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