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Cinnamomum chekiangense somatic embryogenesis and plant regeneration method

A technology of somatic embryos and camphor bodies, applied in plant regeneration, botany equipment and methods, horticultural methods, etc., can solve the problems of prone to differentiation, instability, and inability to fully retain the excellent characteristics of the mother parent in offspring

Active Publication Date: 2020-02-25
ZHEJIANG FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, most of the researches on Cinnamomum camphora in Zhejiang are on seed collection, seedling cultivation, and cold resistance of Cinnamomum camphora in Zhejiang (Wang Nianjin et al., 2003; Hao Riming et al., 2004; Qian Wei et al., 2007; Sun Qimeng et al., 2008; Xie Xiaojin, 2005 ), at present Zhejiang Cinnamomum camphora mainly relies on seed propagation, and the natural seed setting rate of Zhejiang Cinnamomum camphora is low and unstable, which limits its large-scale seedling production and application, and cannot meet the needs of afforestation. At the same time, the offspring of seed propagation are prone to differentiation and cannot completely retain the female parent excellent characteristics of

Method used

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  • Cinnamomum chekiangense somatic embryogenesis and plant regeneration method
  • Cinnamomum chekiangense somatic embryogenesis and plant regeneration method
  • Cinnamomum chekiangense somatic embryogenesis and plant regeneration method

Examples

Experimental program
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Effect test

Embodiment 1

[0025] Isolation of immature zygotic embryos: At the end of August, the immature fruit of a single plant of Cinnamomum chinensis, which grows vigorously and has no pests and diseases, was collected in Hangzhou Botanical Garden. The explants were rinsed under flowing tap water for 2 h. Then, treat it with 75% (w / v) alcohol for 30 seconds on the ultra-clean workbench in the aseptic room, then disinfect it with 0.1% (w / v) mercury chloride for 10-15 minutes, and then rinse it with sterile water for 4-4 minutes. 5 times. Under sterile conditions, the seed coat was peeled off, and the immature zygotic embryos were taken out for the next stage of embryogenic callus induction experiment.

Embodiment 2

[0027] Embryogenic callus induction stage: The immature zygotic embryos obtained in Example 1 were subjected to embryogenic callus induction, using MS medium, and the additional hormone concentrations were 2, 4-D0.05~0.2mg / L and 6 -BA 0.5~2.0mg / L, add 2.0g / L CH and 2.0g / L AC, the medium contains 30g / L sucrose and 7g / L agar. Before sterilization, adjust the pH value of the medium to 5.8, sterilize at 121°C and pressure 101kp for 20 minutes under high temperature and high pressure, and culture at 24±1°C in dark conditions until embryogenic callus is induced.

[0028] Observing the internal morphology of immature fruit embryos of Cinnamomum camphora in different development stages, the fruit development is divided into four stages, that is, the ratio of embryo to seed in the first stage is 1:2 respectively.

[0029] Table 1 Induction rate of embryogenic callus in fruits at different developmental stages

[0030] developmental stage I II III IV Induction rat...

Embodiment 3

[0037]Proliferation stage of embryogenic callus: The embryogenic callus obtained in Example 2 is proliferated, using MS medium, additional hormones as shown in Table 3, adding 2.0g / L of CH, containing 30g / L of CH in the medium L of sucrose and 8g / L of agar, other culture conditions were the same as in the induction stage, the growth of embryogenic callus is shown in Table 3 below.

[0038] Table 3 Effects of different culture media on the growth of embryogenic callus

[0039]

[0040] It can be seen from Table 3 that when 2,4-D 0.2mg / L and 6-BA 0.1mg / L were added to the MS basic medium, the proliferation factor of embryogenic callus was the highest, and the embryogenic callus after proliferation Wounded tissue is light yellow in color, loose in texture, and granular in structure. When the concentration is higher or lower than this concentration, the multiplication factor decreases. Therefore, the optimum proliferation medium is: MS+2,4-D0.05mg / L+6-BA 2.0mg / L+CH 2.0g / L+AC 2...

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Abstract

The invention relates to a Cinnamomum chekiangense somatic embryogenesis and plant regeneration method. The method includes: collecting the fruits of Cinnamomum chekiangense open-pollination half-sibfamilies, and isolating immature zygote embryos to perform somatic embryogenesis culture which comprises an embryonic callus tissue induction stage, an embryonic callus proliferation stage, a somaticembryo maturing stage and a somatic embryo germination stage. The method has the advantages that the embryonic callus induction and culture system of the Cinnamomum chekiangense immature zygote embryos is built for the first time, the optimal embryonic callus tissue induction culture medium and proliferation culture medium are screened out, a good proliferation effect is achieved, a method which is short in cycle, high in breeding rate and low in cost is provided for the large-scale asexual breeding of Cinnamomum chekiangense, and reference is provided for the building of a Cinnamomum chekiangense genetic transformation receptor system which is more efficient.

Description

technical field [0001] The invention relates to the technical field of plant tissue culture and propagation, in particular to a method for embryogenesis and plant regeneration of camphor camphor in Zhejiang. Background technique [0002] Cinnamomum chekiangense is an evergreen broad-leaved tree belonging to the genus Lauraceae. It is a unique plant and a rare tree species in China. It is mainly distributed in East China and scattered or small patches in Zhejiang, Anhui, Jiangxi, Fujian and other places. The amount of resources is relatively small. Less (Sun Qimeng et al., 2008). Zhejiang camphor has excellent material, straight trunk, dense branches and leaves, graceful posture, and bright red fresh leaves. It is an excellent timber and ornamental tree species for greening. At the same time, Cinnamomum camphora has strong resistance to pollutants such as smoke and sulfur dioxide, and can release chemical substances that purify the air and provide health care. 1989). Howev...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 张俊红王晨童再康楼雄珍黄华宏
Owner ZHEJIANG FORESTRY UNIVERSITY