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Ascendens mucronatum somatic embryogenesis method

An embryogenesis and somatic cell technology, applied in the field of forest cell engineering, can solve the problems of long-term continuous production difficulties, long breeding cycle, low seedling emergence rate, etc., and achieve the effect of realizing mass reproduction, meeting market demand and improving induction rate.

Active Publication Date: 2021-08-06
INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Although Zhongshan fir can bear fruit naturally, the emergence rate is extremely low and the breeding cycle is long. Currently, the traditional cutting breeding technology is mainly used for the breeding of improved varieties. However, due to the large restrictions of the natural environment and seasonality, long-term continuous production is very difficult.
Although there are reports on the proliferation of adventitious buds of Zhongshan fir, the proliferation rate is low and rooting is difficult, which is not enough to meet the huge demand of the nursery stock market in the short term.

Method used

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  • Ascendens mucronatum somatic embryogenesis method
  • Ascendens mucronatum somatic embryogenesis method
  • Ascendens mucronatum somatic embryogenesis method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] (1) Preparation of immature zygotic embryos of Zhongshan fir

[0035] From late August to mid-September, the immature cones of Zhongshan fir were collected every 7 days, and quickly stored in a refrigerator at 4°C. Take out the zygotic embryo and observe it under a dissecting microscope. Take the seeds that have developed to the early cotyledon embryo, soak them in 75% ethanol for 45-60 seconds, then sterilize them with 2.5% sodium hypochlorite solution for 18-20 minutes, and finally rinse them with sterile water for 3-60 seconds. 4 times, then under sterile conditions, peel off the hard seed coat outside the seeds with a sterile scalpel and tweezers under a dissecting microscope, take out the immature zygotic embryos, and inoculate them in solid embryogenic callus induction culture Inoculate 8-10 grains per culture dish. Zhongshan fir immature zygotic embryos such as figure 1 shown.

[0036] (2) Induction of embryogenic callus

[0037] Under sterile conditions, imm...

Embodiment 2

[0057] It is basically the same as Example 1, except that the induction solid medium in step (2) is: improved DCR basic medium+NAA 4.0mg / L+6-BA 1.5mg / L+L-Glu 0.3g / L, Pro 0.2g / L+Inositol 0.2g / L+Maltose25g / L+agar 8.5g / L, pH 5.6.

[0058] The maintenance and proliferation medium in step (3) is: 3 / 4 improved DCR basic medium+NAA2.0mg / L+6-BA0.5mg / L+KT 1.0mg / L+Pro 0.2g / L+Inositol 0.2g / L+CH 0.5g / L+Maltose 30g / L+agar 8.5g / L, pH 5.8.

[0059] The somatic embryo induction medium in step (4) is: the improved DCR medium is basic medium+ABA 2.0mg / L+6-furfuryl aminopurine (KT) 2.0mg / L+methyl jasmonate 0.2g / L+polyethylene Diol 8000 (PEG8000) 100g / L+Inositol 0.5g / L+LH 1g / L+Vc 15mg / L+Maltose 20g / L+agar 8.5g / L), pH 6.0. Other each step and control condition are all identical with embodiment 1, and the result shows, also can reach the induction effect of embodiment 1.

Embodiment 3

[0061] Basically the same as Example 1, the difference is that the somatic embryo induction medium in step (4) is: the improved DCR medium is basic medium+ABA 3.0mg / L+6-furfuryl aminopurine (KT) 2.0mg / L+ Methyl jasmonate 0.2g / L+ polyethylene glycol 8000 (PEG8000) 100g / L+Inositol 0.5g / L+LH 1g / L+Vc15mg / L+Maltose 20g / L+agar 8.5g / L), pH 6.0. Other each step and control condition are all identical with embodiment 1, and the result shows, also can reach the induction effect of embodiment 1.

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Abstract

The invention discloses an ascendens mucronatum somatic embryogenesis method, and belongs to the technical field of forest cell engineering. The method comprises the following steps: collecting immature cones of ascendens mucronatum, taking seeds developed to early cotyledon embryos, separating immature zygotic embryos, carrying out induced culture to obtain embryonic calluses containing embryonic suspensor cell clusters, transferring the obtained embryonic calluses into a maintenance and proliferation solid culture medium, carrying out subculture to obtain the embryonic calluses which are proliferated massively, and then transferring the embryonic calluses to a somatic embryo induction culture medium until the ascendens mucronatum somatic embryos are induced. According to the method, the immature zygotic embryos of the ascendens mucronatum are adopted to induce the embryonic calluses, the induction rate is high, a large number of embryonic calluses can be obtained in a short time, the somatic embryos can be induced, and the method is a technology for producing ascendens mucronatum nursery stocks with short period and high efficiency, can relieve the tension of nursery stock demand, realizes the goal of mass propagation of high-quality seedlings, and is the first time in China.

Description

technical field [0001] The invention belongs to the technical field of tree cell engineering, and in particular relates to a method for embryogenesis of somatic cells of Shanshanshan fir. Background technique [0002] The genus Taxodium Rich. belongs to Taxodiaceae, and there are three main species in the genus, Taxodium distichum (Linn.) Rich., T. ascendens Brongn and T. ascendens Brongn. .mucronatum Tenore). Bald cypresses are all tall trees. Due to their deep roots, huge root system, and rapid base expansion, they are widely used in wetland afforestation, windbreak and soil stabilization, and gully filling in Europe and the United States. Zhongshan fir (Taxodium ‘Zhongshanshan’), also known as hybrid cypress, is a general term for excellent clones obtained by crossing three species of cypress, Mexican cypress, and pond fir. Zhongshan fir has the main advantages of the bald cypress tree species: semi-evergreen tall tree, tall and straight trunk, beautiful tree shape, lus...

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 周艳威张凡陈红王传永李乃伟李云龙陆小清蔡小龙种昕冉周婷
Owner INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI