Method for Regenerating Plants from Somatic Embryos of Osmanthus fragrans

A technology for somatic embryos and plant regeneration, applied in the field of plant tissue and cell culture, can solve the problems of difficult organs and the inability of callus to differentiate further, and achieve the effect of high coefficient and short reproductive cycle.

Inactive Publication Date: 2014-10-08
HUAZHONG AGRI UNIV
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  • Description
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Problems solved by technology

[0005] The purpose of the present invention is to establish a kind of highly efficient and rapid osmanthus somatic embryo regeneration method for the current osmanthus fragrans tissue culture process in which the callus cannot be further differentiated and the organ generation is difficult, so as to provide a new method for industrial seedling cultivation and artificial seed preparation in the future ways and techniques

Method used

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  • Method for Regenerating Plants from Somatic Embryos of Osmanthus fragrans
  • Method for Regenerating Plants from Somatic Embryos of Osmanthus fragrans
  • Method for Regenerating Plants from Somatic Embryos of Osmanthus fragrans

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Experimental program
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Effect test

Embodiment 1

[0034] Embodiment 1: the screening of osmanthus zygotic embryo sampling site

[0035] The zygotic embryo of the osmanthus tree of the test material was taken from the fruit of the osmanthus tree Jingui (the variety is a kind of osmanthus tree commonly cultivated in Wuhan, Hubei Province, China) that grows robustly and is more fruitful on the campus of Huazhong Agricultural University (see figure 2 a,b in). The sampling time was in the morning of fine weather in March. After the fruit was harvested indoors, add a small amount of washing powder and wash it with tap water for 1~2 hours, remove the outer testa and pulp for disinfection, and put it in an ultra-clean workbench with 75% (V / V) sterilized alcohol for 30 seconds, rinsed twice with sterile water, then disinfected with 0.1% (W / V) mercuric chloride for 10 minutes, and rinsed three times with sterile water.

[0036] Will complete osmanthus zygotic embryo (see figure 2 c) were cut into cotyledons respectively (see figur...

Embodiment 2

[0040] Embodiment 2: the screening of sweet-scented osmanthus zygotic embryo best development state

[0041] The zygote embryos for the test were taken from the fruit of the vigorous and fruitful osmanthus tree "Jingui: "Jingui: fruit (this variety is the osmanthus tree variety commonly cultivated in Wuhan, Hubei Province, China). In the morning of the weather, the materials are collected every 30 days or so (in case of rainy weather, it can be adjusted appropriately), until the fruit is fully mature and falls off naturally in May.

[0042] Remove the outer testa of sweet-scented osmanthus seed, pulp and carry out disinfection, and disinfection method is with embodiment 1. Use zygotic embryos of different development states as explants (Table 1), cut 1-3mm fragments on the ultra-clean workbench and inoculate them in MS in time, add 0.5mg / L 2,4-dichlorophenoxyacetic acid ( 2,4-D), 1.0mg / L 6-benzylaminoadenine (6-BA), 30g / L sucrose and 7g / L agar, in a callus induction medium wi...

Embodiment 3

[0047] Embodiment 3: Osmanthus fragrans embryogenic callus induction medium screening

[0048] The zygote embryos of the test material were taken from the Jingui fruit of the strong and strong Osmanthus fragrans tree in the campus of Huazhong Agricultural University. The materials were collected in the morning of fine weather in March, and the outer testa and pulp were removed after the fruit was collected indoors and then disinfected. The specific disinfection method was the same as in Example 1 above. The embryos of the sterilized seeds were completely peeled off, cut into small fragments of 1-3mm on the ultra-clean workbench, inoculated in MS respectively, and added 0~2.5mg / L 2,4-dichlorophenoxyacetic acid ( 2,4-D), 0~2.5mg / L 6-benzylaminoadenine (6-BA), sucrose 30g / L and agar 7g / L, medium pH 5.8~6.5 for induction (Table 2), cultured in the dark at 25°C for 30 days (the same conditions as in Example 1), to obtain embryogenic callus.

[0049] Table 2 Effects of different h...

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Abstract

The invention belongs to the technical field of plant tissue and cell culture, and particularly relates to a method for a regeneration plant of a somatic embryo of osmanthus fragrans Lour. The method takes zygotic embryo of the osmanthus fragrans Lour as an explant, an osmanthus fragrans Lour regeneration system is built through a body cell embryo generating way so as to solve the problem that a callus can not further differentiate by an osmanthus fragrans Lour tissue culture technology. The method comprises the following steps: taking the zygotic embryo of the osmanthus fragrans Lour as materials; building the osmanthus fragrans Lour somatic embryo regeneration system through five stages: embryonic callus induction, the differentiation of the embryonic callus into somatic embryos, somatic embryos maturity and germination, somatic embryos rootage and seedling of the somatic embryos regeneration plant. According to the invention, through comparing different development states of the zygotic embryo and the different using parts of the zygotic embryo, the development part and state suitable for explant are determined. Due to the adoption of the method, a large number of the regeneration plants of the somatic embryo of osmanthus fragrans Lour, which have the consistent genetic basis, can be obtained, so that a novel method is provided for preservation of sources of the rear osmanthus fragrans Lour varieties, industrialized production of good genetype nursery stocks, and preparation of artificial seeds.

Description

technical field [0001] The invention belongs to the technical field of plant tissue and cell culture, and specifically relates to a method for plant regeneration of osmanthus fragrans somatic embryos. Using osmanthus zygotic embryos as explants, an osmanthus fragrans regeneration system is established through somatic embryogenesis to solve the problem of osmanthus fragrans tissue culture. The problem of the inability of the injured tissue to differentiate further. Background technique [0002] Sweet osmanthus (Osmanthus fragrans Lour.) belongs to Oleaceae (Oleaceae) Osmanthus (Osmanthus), originated in Asia, is one of the traditional ten famous flowers with Chinese characteristics. After the 1870s, it began to spread to Europe, the United States, Japan, Korea, and India, and it has a cultivation history of more than 2,000 years in my country. Osmanthus fragrans is beautiful in shape, evergreen in all seasons, and pleasant in aroma. It is known as "exclusively in three autum...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 王彩云邹晶晶高微蔡璇杨谢寒
Owner HUAZHONG AGRI UNIV
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