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Method for inducing embryogenic callus and proliferation by utilizing immature embryos of taxodium 'zhongshanshan'

An embryogenic callus, immature embryo technology, applied in the field of forest cell engineering, can solve the problems of low proliferation rate, long breeding cycle, low seedling emergence rate, etc.

Inactive Publication Date: 2021-03-12
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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  • Method for inducing embryogenic callus and proliferation by utilizing immature embryos of taxodium 'zhongshanshan'
  • Method for inducing embryogenic callus and proliferation by utilizing immature embryos of taxodium 'zhongshanshan'
  • Method for inducing embryogenic callus and proliferation by utilizing immature embryos of taxodium 'zhongshanshan'

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] (1) Preparation of immature zygotic embryos of Zhongshan fir

[0029] 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 wash 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. immature zygotic embryos of Zhongshan fir figure 1 shown.

[0030] (2) Induction of embryogenic callus

[0031] Under sterile conditions, immature ...

Embodiment 2

[0047] It is basically the same as Example 1, except that the induction solid medium in step (2) is: improved DCR basic medium+NAA 3.0mg / L+6-BA 1.0mg / L+L-Glu 0.3g / L, Pro 0.2g / L+Inositol 0.2g / L+Maltose25g / L+agar 8.5g / L, pH 5.6.

[0048] The maintenance and proliferation medium in step (3) is: 3 / 4 improved DCR basic medium+NAA2.0mg / L+6-BA0.2mg / 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. Other steps and control conditions are the same as in Embodiment 1, and the experimental results show that the induction effect of Embodiment 1 can also be achieved.

Embodiment 3

[0050] 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.

[0051] The maintenance and proliferation medium in step (3) is: 3 / 4 improved DCR basic medium+NAA 1.0mg / 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. Other steps and control conditions are the same as in Embodiment 1, and the experimental results show that the induction effect of Embodiment 1 can also be achieved.

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Abstract

The invention discloses a method for inducing embryogenic callus and proliferation by utilizing immature embryos of taxodium 'zhongshanshan', and belongs to the technical field of forest tree cell engineering. The method comprises the following steps of collecting immature cones of taxodium 'zhongshanshan', taking seeds developed to early cotyledon embryos, separating out immature zygotic embryos,carrying out induction culture to obtain the embryogenic callus containing embryogenic suspensor cell clusters, transferring the obtained embryogenic callus into a maintenance and proliferation solidculture medium, and carrying out subculture to obtain a large amount of proliferated embryogenic callus. The method is not limited by seasons and external environments, the embryogenic callus can beefficiently induced, a large number of white and compact embryogenic callus can be obtained in the maintenance and proliferation culture medium in a short time, an efficient way is provided for rapidpropagation of excellent taxodium 'zhongshanshan' strains, preservation of excellent germplasm resources is facilitated, an early-stage basis is provided for subsequent regeneration plants and a genetic transformation receptor system, the goal of mass propagation of high-quality seedlings can be achieved, and the market requirements are greatly met.

Description

technical field [0001] The invention belongs to the technical field of tree cell engineering, and specifically relates to a method for inducing embryogenic callus and proliferating by using immature embryos of Zhongshan 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 stra...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/005
Inventor 周艳威张凡陈红王传永李云龙陆小清李乃伟蔡小龙种昕冉周婷
Owner INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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