Zhejiang phoebe somatic embryo induction method

A technology of somatic embryos and cell embryos, which is applied in the field of Zhejiang Nan somatic embryo induction, can solve the problems of inability to realize large-scale breeding applications, failure to survive, and low survival rate

Active Publication Date: 2017-08-18
ZHEJIANG FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the survival rate of cutting propagation of Phoebe zhejiang has been low so far, and due to the limitation of cutting materials and seasons, large trees cannot survive due to their physiological age, and large-scale breeding applications cannot be realized.

Method used

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  • Zhejiang phoebe somatic embryo induction method
  • Zhejiang phoebe somatic embryo induction method
  • Zhejiang phoebe somatic embryo induction method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Induction of somatic embryogenesis of immature zygotic embryos

[0020] From mid-July to late August, the immature fruits on a single plant of Panshan Phoebe, which grow robustly and are free from diseases and insect pests, were harvested.

[0021] First rinse in running tap water for 2 hours. Treat with 75% alcohol on the ultra-clean workbench in the aseptic room for 30 seconds, rinse with sterile distilled water for 3 to 4 times, sterilize with 0.1% mercuric chloride for 8 minutes, 10 minutes, 12 minutes and 14 minutes, and then rinse with sterile distilled water for 5 minutes. ~7 times to wash off residual mercury chloride. After mercuric chloride treatment at different times, the effects on explants were significantly different, as shown in Table 1.

[0022] Table 1 Effect of different mercuric chloride treatment time on explant disinfection

[0023] Mercury treatment time Disinfection effect explant state 8min Incomplete, bacteria r...

Embodiment 2

[0034] Embodiment 2 The influence of different developmental stages of immature embryos on the incidence rate of somatic embryos

[0035] On July 17, July 24, July 31, August 7, August 14, and August 21, 2016, respectively, 150 seeds of 4 excellent plants were collected, of which 135 were used for body For the embryogenesis test, the other 15 were used for dissection observation, and the developmental stages of the zygotic embryos at each collection time were analyzed. The seeds of 4 superior plants were recorded as A, B, C, and D respectively. The peeled immature embryos were placed in 0.5M sucrose, pretreated at 4°C for 72 hours, and inoculated in BIM medium, namely MS+CH500mg / L+glutamine 500mg / L+6-BA 1.0mg / L+2,4-D0.5mg / L was used for somatic embryogenesis experiment to determine the influence of different collection time, that is, different development stages of zygotic embryos, on the somatic embryogenesis rate. Concrete operation steps are with embodiment 1.

[0036] Ta...

Embodiment 3

[0039] Embodiment 3 Proliferation of somatic embryos

[0040] The induced somatic embryos are placed in a proliferation medium for proliferation. The number of somatic embryos in each treatment was 25, 5 in each dish, 5 dishes in total, and 3 repetitions were set for each treatment. The culture temperature was 25°C±2°C, and the culture was carried out in the dark.

[0041] Table 5 Effects of different proliferation media on the proliferation of somatic embryos

[0042] 1 2 3 4 5 6 6-BA 0 0 0 1.0 1.0 1.0 NAA 0.1 0.5 1.0 0.1 0.5 1.0 Multiplication coefficient 4.8±1.3 14.3±2.4 7.9±2.0 5.2±1.9 6.4±2.3 4.0±1.4

[0043] The induced primary embryos were cultured in the proliferation medium, and after 3 weeks, a white embryo-like structure similar to that of the parent was observed on the surface of the primary embryos, which was a secondary embryo obtained through secondary embryogenesis. Prolonging the time of proliferation cul...

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Abstract

The invention discloses a Zhejiang phoebe somatic embryo induction method which comprises the following steps: 1) performing surface sterilization on immature fruits of Zhejiang phoebe as explants; 2) peeling off young embryos of seeds, inoculating with a somatic embryo induction culture medium, and performing somatic embryo induction; and 3) inoculating the induced somatic embryos of the step 2) with the somatic embryo enrichment medium to proliferate somatic embryos to generate secondary embryos. By adopting the Zhejiang phoebe somatic embryo induction method, a somatic embryo direct generation method for immature zygotic embryos of the Zhejiang phoebe is established for the first time, and the somatic embryo generation rate can be up to 65% on the culture medium of the method with the zygotic embryos which are at the stage of globular embryos or are converted into heart-shaped embryos. On an optimal enrichment culture medium, the enrichment coefficient can be up to 14.3, and on an optimal somatic embryo germination culture medium, the germination rate of the somatic embryos can be up to 68.5%.

Description

technical field [0001] The invention belongs to the field of plant biotechnology, and in particular relates to a method for inducing somatic embryos of Phoebus zhejiang. Background technique [0002] Phoebe chekiangensis is a tall tree belonging to the genus Phoebe in Lauraceae. It is one of the original planting species of "Golden Phoebe". It has high economic value and is a unique and precious tree species in the eastern subtropical zone. It is naturally distributed in Zhejiang and northern Fujian , Northeastern Jiangxi and Southern Anhui (Editorial Committee of Flora of China, Chinese Academy of Sciences, 1982; Collaborative Group of Flora of Anhui, 1986). The trunk is straight and the material is beautiful. It is a special material for high-end furniture. The tree is tall and green in all seasons. It is also an excellent tree species for landscaping. Although Zhejiang Phoebe has high economic and ornamental value, there are very few existing resources, and the protectio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/00A01H4/001
Inventor 童再康张俊红师晶晶
Owner ZHEJIANG FORESTRY UNIVERSITY
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