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Aesculus hippocastanum somatic embryogenesis and strain regenerating method

A technique for embryogenesis and horse chestnut, applied in the directions of plant regeneration, botanical equipment and methods, horticultural methods, etc., can solve the problems of inability to meet the needs of European horse chestnut, slow reproduction, consuming manpower, material and financial resources, etc.

Inactive Publication Date: 2004-09-15
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0021] In view of the huge medicinal and ornamental value of horse chestnut, the demand for horse chestnut in the domestic market is increasing. At present, the introduction and cultivation of horse chestnut in China is limited, and its propagation method is limited to sowing, and the reproduction speed is extremely fast. Slow, and extremely consuming manpower, material and financial resources, far from meeting the market's demand for horse chestnut

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1: Tissue culture using germs of seeds

[0068] The seed coat of horse chestnut is thick and smooth, which is difficult to cut, so it can be germinated first. When germinating, the radicle first protrudes from the seed coat, and after 5 days, the exposed part of the seed coat can reach 4cm. This part is covered with germs. Select the part that does not expose the germ, wash it with tap water for 2 hours, and use 75% of the seed coat on the ultra-clean workbench. Soak in alcohol for 30s, 0.1% HgCl 2 Soak for 8-12 minutes, rinse with sterile water for 5-6 times, blot the surface moisture with sterile filter paper, remove the outer covering of the germ, take out the germ, and inoculate it in MS to grow for a period of time. Because the seed coat is dense, the cotyledons are thick, and the germ is not easily polluted by the outside world. Skilled people can also directly remove the outer packaging of the germ without using disinfectant to inoculate it, so as to avo...

Embodiment 2

[0089] Embodiment 2: Adopt the leaf of the adventitious bud that spring branch just pulls out or is induced by germ to carry out tissue culture

[0090] In this embodiment, there are two ways for the source of explants. One is to take the leaves from the aseptic seedlings obtained in Example 1 and inoculate them on the induction medium, which can save the step of disinfection; Materials are collected from germinated branches, and young leaves (with petioles) with bright green color and good growth state are selected, soaked in detergent for 2 hours, rinsed with running water for 2 hours, and disinfected on an ultra-clean workbench. During disinfection, each triangular bottle can be put into 4 to 6 leaves, 75% absolute ethanol for 1 min, 0.1% 84 disinfectant for 8 min, rinse with sterile water for 5 to 6 times, wash the surface with sterile filter paper, cut off the edge of the leaf, and cut into 5mm×10mm and cut 3 times across the midrib of the leaf, then put the backside down...

Embodiment 3

[0138] Example 3: Tissue culture with cotyledons after somatic embryo growth for half a month

[0139] The somatic embryo of the European horse chestnut is quite special. The cotyledons cannot develop into leaves like the somatic cotyledons of other plants, but gradually crack during the growth of the somatic embryo, which is very hypertrophic, and is the induction of adventitious buds and somatic embryogenesis again. good material. In this example, MS is the minimal medium, the hormones used are ZT, BA, KT, NAA, IAA and a new type of cytokinin TDZ was tried. The cotyledons of somatic embryos were cut directly and inoculated on the prepared medium. It was found that the cotyledons produced adventitious buds in the form of direct organogenesis, and somatic embryos in two forms of direct and indirect organogenesis. Different hormone combinations had a great influence on cotyledon direct organogenesis to produce adventitious buds or somatic embryos, as shown in Table 8.

[0140...

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Abstract

The present invention provides a seedling cultivation method of aesculus hippocastanum with high ornamental and medicinal value. It is characterized by that it uses seed plumula of aesculus hippocastanum or new bud germinated by spring plant, leaf cultured by seed plumula and cotyledon formed after embryo proper is grown for 15 days to make tissue cultivation method of somatic embryogenesis and plant regeneration so as to implement said invention.

Description

1. Technical field [0001] The invention belongs to cell embryogenesis and clone plant regeneration in tree tissue culture technology in forestry science [0002] technology field. 2. Background technology [0003] There are 2 existing genera of Hippocastanaceae in the world, among which there are about 30 species of Aesculus L, which are mainly distributed in the northern temperate zone. There are about 10 species in my country, which are mainly distributed in the subtropical regions in the southwest. They are all tall deciduous trees, usually 20-30m high, and the highest can reach 45m. Among them, horse chestnut, long-handled horse chestnut, Zhejiang horse chestnut, Yunnan horse chestnut, Tianshi chestnut, Japanese horse chestnut, etc. are widely distributed in my country, and Shaanxi, Henan, Jiangsu, Zhejiang and other places are all It has been cultivated, and there is still an ancient tree more than 300 years old in Longdong, Gansu. Due to climate change, the distribut...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 施季森吕秀立
Owner NANJING FORESTRY UNIV
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