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Embryogenic calli induction and proliferation method of pinus elliottii engelm

A technology of embryogenic callus and slash pine, which is applied in the field of plant tissue culture, can solve the problems of high cost, difficulty in meeting production requirements, and low efficiency, and achieve the effects of increasing yield, simple steps, and short culture time

Inactive Publication Date: 2019-07-16
JIANGXI ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the research on vegetative propagation technology of slash pine includes cutting, grafting, conventional tissue culture and other methods, but the cost is high, the efficiency is low, and it is difficult to meet the production requirements

Method used

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  • Embryogenic calli induction and proliferation method of pinus elliottii engelm
  • Embryogenic calli induction and proliferation method of pinus elliottii engelm
  • Embryogenic calli induction and proliferation method of pinus elliottii engelm

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A method for inducing and proliferating embryogenic callus of pine slats, comprising the following steps:

[0026] 1. Selection of Explants

[0027] At the end of June, natural pollinated immature cones were taken from the pine trees in the seed orchard of the Xiajiang Forest Tree Improved Seed Farm in Ji'an City, with good growth and no pests and diseases. They were stored in an ice box on the day of collection and brought back to the laboratory refrigerator at 4°C. save. Before use, rinse the cones with water to remove impurities such as stains on the cones, absorb the water with filter paper, and then perform the following steps on a sterile operating table: after immersing the cones in 75% alcohol for 5 minutes, rinse them once with sterile water, Soak in 0.1% mercuric solution for 10 minutes, take out and rinse with sterile water 3-4 times with filter paper to absorb water, then peel off the immature seeds, gently remove the outer epidermis of the seeds with a sca...

Embodiment 2

[0041] Example 2: The effect of different basic media on the induction of embryogenic callus from immature embryos of pine pine

[0042] According to the operation method of Example 1, the explants were inoculated on different basic mediums to induce proliferation, and the results of the induction of embryogenic callus of pine slats by the basic medium DCR, WPM, MS, B5 and 1 / 2MS were tested. As shown in table 2.

[0043] Table 2 shows: in DCR medium, the induction rate of embryogenic callus induced by immature seeds of slash pine is the highest, reaching 30.65%; followed by WPM, MS and B5 medium, the induction rate of embryogenic callus is 25.40% and 14.29%, respectively. %, 7.14%; while the induction rate was the lowest in 1 / 2MS medium, and the induction rate was 0. From the test results, we believe that the minimal medium is essential for inducing embryogenic callus differentiation of immature seeds. Comparing the five basic media, it was found that the induction culture o...

Embodiment 3

[0046] Example 3: Effects of Different Hormones on Induction Efficiency of Embryogenic Callus from Immature Seeds of Pine

[0047] In the same DCR basic culture and adding sucrose 30g / L, carrageenan 7g / L, L-glutamine 300mg / L, hydrolyzed casein 500mg / L, inositol 1g / L, 2-(N-morpholine)ethanesulfonate Under the condition of acid monohydrate 250mg / L, the comparison of different hormones and their combinations on the induction of embryogenic callus from immature embryos of sambar pine was tested. The results are shown in Table 3.

[0048] Table 3 shows that no embryogenic callus was induced in the medium without any hormones; only 2,4-D was induced in the medium with single addition of hormones 6-BA, KT, TDZ and 2,4-D Embryogenic callus formation. When 2,4-D was used in combination with 6-BA and KT, the induction rate of embryogenic callus was significantly higher than that when each hormone was used alone. The combination of 2,4-D and KT was significantly better than the combinat...

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Abstract

The invention provides an embroyogenic calli induction and proliferation method of pinus elliottii engelm. The method includes the steps of 1), induced culture of embroygenic calli, namely placing immature seed embryos of the pinus elliottii engelm in an embroyogenic calli induction medium (with a DCR culture medium as a basic culture medium added with, by weight, 30g / L of saccharose, 7g / L of carrageenan, 1.0mg / L-2.0mg / L of 2,4-dichlorophenoxyacetic acid (2,4-D), 1.0mg / L-2.5mg / L of N6-furan methyl adenine (KT), 300mg / L of L-glutamine, 250mg / L-500mg / L of casein hydrolysate, 1g / L of inose and 250mg / L of 2-(N-morpholine) ethylsulfonic acid monohydrate) for induced culture to obtain primary embryogenic calli; 2), proliferation of the embryogenic calli, namely transferring the embryogenic callli obtained in the step 1) into a proliferation culture medium (with the DCR culture medium as the basic culture medium added with, by weight, 30g / L of maltose, 7g / L of carrageenan, 0.5mg / L-1.0mg / L of2,4-D, 0.5mg / L-1.0mg / L of 6-benzylamino adenine (6-BA), 300mg / L of L-glutamine and 250mg / L-500mg / L of casein hydrolysate for proliferation culture to obtain proliferating embryogenic calli.

Description

technical field [0001] The invention relates to the technical field of plant tissue culture, in particular to a method for inducing and proliferating embryogenic callus of pine pine. Background technique [0002] Swamp pine (Pinus elliottii Engelm) is a tall tree native to the low-altitude and warm regions of the southeastern United States. It has been widely planted in Jiangxi, my country due to its excellent characteristics of barren tolerance, fast growth, good wood properties and high turpentine yield. , Fujian, Hunan and other southern provinces. Due to its high economic and ecological value, pine pine has become one of the main tree species in my country's artificial forest management. For many years, scientific researchers have been continuing research on the genetic improvement of slash pine, but the traditional multi-generational cross-breeding cycle of slash pine is long, which seriously restricts the breeding of fine pine varieties. How to shorten the breeding cyc...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/005
Inventor 程强强杨春霞丁伟谷振军肖复明杜强
Owner JIANGXI ACAD OF FORESTRY
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