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Germplasm test tube storage and ex-situ conservation method for East Asian phyllitis japonica

A preservation method and germplasm technology, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve problems such as scarcity of reserves, degradation of varieties, damage to fragile ecological environment, etc.

Inactive Publication Date: 2013-04-03
JILIN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Artificial reproduction requires a large amount of germplasm, but the natural reserves of the East Asian folio fern are scarce, and a large amount of wild germplasm is collected. This behavior itself will cause damage to the already fragile ecological environment, so that the damage is greater than the consequences of protection.
Artificial long-term subculture will also cause the species to degenerate, which is not suitable for growth and reproduction in the wild environment.

Method used

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  • Germplasm test tube storage and ex-situ conservation method for East Asian phyllitis japonica
  • Germplasm test tube storage and ex-situ conservation method for East Asian phyllitis japonica
  • Germplasm test tube storage and ex-situ conservation method for East Asian phyllitis japonica

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Spore Germination

[0029] The material comes from the forest area of ​​Baishan City, Jilin Province; the sporophyll containing mature sporangia is taken, and the sporophyll is cut into 1 cm in size in the laboratory; 0.1% HgCl under sterile conditions 2 The solution was sterilized for 5 minutes, rinsed with sterile water 6 times, blotted dry with filter paper and inoculated.

[0030] Inoculation method: There are three methods of inoculation, one is direct inoculation of sporophylls; the other is inoculation after stripping the sporangia in the sporophylls under the dissecting microscope of the ultra-clean workbench; On the filter paper with the size of 2cm × 2cm after bacteria, carefully crush the sporangia with a scalpel, release the spores in it, and inoculate together with the small filter paper.

[0031] Spore culture: Two methods of solid culture and liquid culture of 1 / 2MS+20g of sucrose were adopted.

[0032] Culture conditions : 12h / d under...

Embodiment 2

[0035] Example 2 Prothallus Subculture and Proliferation

[0036] Cut the prothallus formed after spore germination into small pieces of about 0.5 cm in size, each small piece contains about 5-10 pieces of prothallus, and inoculate them into solid MS medium containing different concentrations of plant growth regulators (See Table 1), inoculate 8-10 pieces per bottle.

[0037] The protophyll mass formed after spore germination was respectively inoculated into MS medium containing different concentrations of plant growth regulators, culture conditions: 100-200lx scattered light, light 12h / d. The daytime temperature is 25-26°C, and the nighttime is 20-22°C, which is the same as the spore germination condition; after 60 days, observe the proliferation and growth of the prothallus (see Table 1, Image 6 , Figure 7 ).

[0038] Prothallus can proliferate at a rate of about 5 times in several media tested. The new multiple range test showed that there was no significant diffe...

Embodiment 3

[0040] Embodiment 3 prothallus test tube preservation

[0041] will grow robust prothalloids ( Image 6 ) placed in a light incubator. The culture temperature is 10, 12, 14, 15, 16, and 17°C; the light is 5-7h / d; the light intensity is 200-300lx; the medium is 1 / 2MS+sucrose 25-30g / L, or MS+sucrose 25-30g / L of solid medium; once every 12 months, a part of the prothallus was kept for preservation and culture during each transfer, a part was transferred to normal culture conditions for proliferation and subculture treatment, and a part was used for sporophyte induction outside the test tube. Observe its subculture proliferation and sporophyte plant formation.

[0042]After the prothallus were cultured at 14, 15, and 16°C for 12 months, there was almost no growth and proliferation, and the state was similar to that at the beginning of the culture. After being transferred to subculture conditions, it can still proliferate and grow normally; After 60 days of cultivation, the v...

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Abstract

The invention provides a germplasm storage method for East Asian phyllitis japonica, which comprises the following steps that: prothallus is inoculated in a 1 / 4-MS (full) + sucrose 25-30 g / L medium, and the medium is placed in an illumination incubator, wherein the illumination intensity is 200-300 lx, the illumination is 5-7 h / d, and the temperature is 14-16 DEG C; 12 months later, almost no growth and proliferation occurs, and the state of the prothallus is similar as that in initial culture, and after the prothallus is transferred into successive culture conditions, the prothallus can proliferate and grow normally; and the prothallus is subjected to five-year storage treatment. The invention also provides an ex-situ conservation method for East Asian phyllitis japonica, which is implemented through a way that the stored prothallus is subjected to sub-cultured reproduction every year, and the formation and ex-situ transplanting of sporophore plants are induced by taking turfs as a matrix outside a test tube; three sits with basically similar as the original growing environment of phyllitis japonica are selected as ex-situ transplanting test areas in Changbai Mountains; 100 plants are planted at each site; and after growing 4 years, the growth vigour of the prothallus is good, thereby truly realizing the protection on endangered plants.

Description

technical field [0001] The invention belongs to the field of biological technology, in particular to a germplasm preservation method and ex situ breeding protection method of the endangered and rare plant Pteridophyllum japonicus. Background technique [0002] East Asian Aloe Fern ( Phyllitis japonica Kom.) is also known as Northeast folio fern, Japanese folio fern, and folio fern. It has both ornamental value and medicinal value. It is a perennial evergreen herbaceous plant belonging to the family Aeridaceae. Rhizomes short, recumbent or oblique, roots slender, mostly horizontal. Rare species in the world, only one genus and one species in my country, national second-class rare and endangered protected plants. Produced in some areas on the south and west sides of Changbai Mountain, it is mainly distributed in some narrow areas such as Changbai Korean Autonomous County, Linjiang, Ji'an, Fusong, etc. in Jilin Province, and the distribution is scattered. Folio fern like...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 赵春莉赵和祥顾德峰李金英黄翔童陈丽飞董然刘晓嘉刘淑英姜立新
Owner JILIN AGRICULTURAL UNIV
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