CN109430060B - Culture method for prolonging storage life of kiwi fruit tissue culture seedlings - Google Patents
Culture method for prolonging storage life of kiwi fruit tissue culture seedlings Download PDFInfo
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- CN109430060B CN109430060B CN201811603447.2A CN201811603447A CN109430060B CN 109430060 B CN109430060 B CN 109430060B CN 201811603447 A CN201811603447 A CN 201811603447A CN 109430060 B CN109430060 B CN 109430060B
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- tissue culture
- culture
- kiwi fruit
- seedlings
- storage life
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- 244000298697 Actinidia deliciosa Species 0.000 title claims abstract description 27
- 235000009436 Actinidia deliciosa Nutrition 0.000 title claims abstract description 27
- 238000012136 culture method Methods 0.000 title claims abstract description 9
- 239000012879 subculture medium Substances 0.000 claims abstract description 9
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims abstract description 7
- 229930006000 Sucrose Natural products 0.000 claims abstract description 7
- 238000000034 method Methods 0.000 claims abstract description 7
- 239000005720 sucrose Substances 0.000 claims abstract description 7
- 238000005286 illumination Methods 0.000 claims description 8
- 229920001817 Agar Polymers 0.000 claims description 4
- 239000008272 agar Substances 0.000 claims description 4
- 230000004083 survival effect Effects 0.000 abstract description 4
- 230000014759 maintenance of location Effects 0.000 abstract 3
- 239000001963 growth medium Substances 0.000 description 9
- 238000004321 preservation Methods 0.000 description 7
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 238000004161 plant tissue culture Methods 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 2
- 241000220324 Pyrus Species 0.000 description 2
- 229930003268 Vitamin C Natural products 0.000 description 2
- 230000003111 delayed effect Effects 0.000 description 2
- 238000012423 maintenance Methods 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 230000035764 nutrition Effects 0.000 description 2
- 235000021017 pears Nutrition 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 235000019154 vitamin C Nutrition 0.000 description 2
- 239000011718 vitamin C Substances 0.000 description 2
- 244000144730 Amygdalus persica Species 0.000 description 1
- 244000141359 Malus pumila Species 0.000 description 1
- 244000183278 Nephelium litchi Species 0.000 description 1
- 235000006040 Prunus persica var persica Nutrition 0.000 description 1
- 235000021016 apples Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 231100000957 no side effect Toxicity 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a culture method for prolonging the retention period of a tissue culture seedling of kiwi fruit, which is characterized in that a certain amount of sucrose is added on the basis of a traditional subculture medium of the tissue culture seedling of kiwi fruit to effectively prolong the retention period of the tissue culture seedling and solve the problem of short retention period of the tissue culture seedling of kiwi fruit at present. The method of the invention can generally delay the growth of the tissue culture seedling for 4-5 months, and the survival rate of the tissue culture seedling can reach more than 80%.
Description
Technical Field
The invention belongs to the technical field of plant tissue culture, and particularly relates to a culture method for prolonging the storage life of tissue culture seedlings of kiwi fruits.
Background
The kiwi fruit is rich in nutrition, contains various vitamins, fat, protein and other components, wherein the content of vitamin C is 100 mg-400 mg/100 g, which is far higher than that of oranges, pears and apples, and the vitamin C is called as Vc king. Besides the edible value, the kiwi fruit has beautiful and fragrant flowers and higher ornamental value. China is the center of kiwi fruit resource distribution, germplasm resources are rich and widely distributed, but the kiwi fruit is in a wild or semi-wild state for a long time, and some species are in endangered places due to deterioration of ecological environment, so that collection and storage are urgently needed.
The common preservation method is to collect seeds, branches and seedlings to establish a resource garden. The resource garden building has the defects of large occupied space, large labor, high maintenance cost and the like. The appearance, development and perfection of the plant tissue culture technology provide possibility for the in vitro preservation of plant germplasm resources. The in vitro preservation has the advantages of sterile culture, small occupied space, low labor and maintenance cost, convenience for germplasm exchange in various regions, rescue and rapid propagation of endangered species and the like, and is applied to various plants (peaches, pears, litchis and the like). Therefore, the preservation of the germplasm resources of the kiwi fruits by utilizing the plant tissue culture technology is very necessary and urgent.
In the process of preserving the germplasm of the plant in vitro, the nutrient components and water in the culture medium are gradually consumed, and the culture is inoculated to a new culture medium at intervals so as to ensure the normal growth. The tissue culture seedlings of the kiwi fruits are subcultured once in 1 month generally, can be stored for about 2 months at most, and cost a large amount of manpower, material resources and time for frequent subculture; meanwhile, with the increase of the number of subcultures, the mutation probability of the culture material is increased, which is not favorable for the preservation of the original germplasm.
Disclosure of Invention
The invention aims to provide a culture method for prolonging the storage life of tissue culture seedlings of kiwi fruits, and provides technical support for in vitro preservation of kiwi fruit germplasm resources by using a plant tissue culture technology.
In order to realize the purpose, the invention adopts the following technical scheme:
a culture method for prolonging the storage life of tissue culture seedlings of kiwi fruits comprises the following steps:
(1) inoculating the non-pollution and strong-growing kiwi fruit tissue culture seedlings into a special subculture medium;
(2) preserving in a culture chamber with the culture temperature of 25 +/-1 ℃, the illumination time of 12 h/d and the illumination intensity of 1500-2000 lx;
(3) when the leaves of the tissue culture seedlings are dry, the new special subculture medium is replaced.
The special subculture medium is MS +6-BA 1.0-2.0 mg/L + NAA 0.5-1.0 mg/L + GA30.1 mg/L + agar 7g/L + sucrose 15-20 g/L.
The invention has the beneficial effects that:
(1) according to the method, the growth of the tissue culture seedlings is effectively slowed down by limiting the concentration of sucrose in the culture medium, the nutrition consumption is reduced, the subculture interval time of the kiwi fruit tissue culture seedlings is prolonged from 2 months to 4-5 months, the subculture workload is greatly reduced, the survival rate of the tissue culture seedlings can reach more than 80%, and the mutation probability is also reduced.
(2) The method has the characteristics of simplicity, easiness in operation, safety, reliability, no side effect on the culture material, simplicity, effectiveness, strong operability, high application value and the like.
TABLE 1 influence of different sucrose concentrations on the shelf life of tissue culture seedlings of Kiwi fruit
As can be seen from the results in Table 1, the preservation period of the kiwi fruit tissue culture seedlings can be doubled by adopting the method, and the tissue culture cost is saved.
Detailed Description
In order to make the present invention more comprehensible, the technical solutions of the present invention are further described below with reference to specific embodiments, but the present invention is not limited thereto.
Example 1
A culture method for prolonging the storage life of tissue culture seedlings of kiwi fruits comprises the following steps:
(1) according to the formula of MS +6-BA 1.0 mg/L + NAA 0.5 mg/L + GA3Preparing a special subculture medium from 0.1 mg/L agar 7g/L sucrose 15 g/L; adjusting the pH value to 6.0 before the culture medium is sterilized;
(2) selecting a pollution-free and strong-growing kiwi fruit tissue culture seedling to inoculate in the culture medium;
(3) placing the inoculated kiwi fruit tissue culture seedling in a culture room with the culture temperature of 25 +/-1 ℃, the illumination time of 12 h/d and the illumination intensity of 1500-;
(4) the tissue culture seedling with delayed growth can be maintained for 5 months, and when the leaf of the tissue culture seedling is found to be dry, the new culture medium is replaced in time. The survival rate of the cultured tissue culture seedling reaches 86 percent.
Example 2
A culture method for prolonging the storage life of tissue culture seedlings of kiwi fruits comprises the following steps:
(1) according to the formula of MS +6-BA 1.0 mg/L + NAA 1.0 mg/L + GA3Preparing a special subculture medium from 0.1 mg/L agar 7g/L and sucrose 20 g/L; adjusting the pH value to 6.0 before the culture medium is sterilized;
(2) selecting a pollution-free and strong-growing kiwi fruit tissue culture seedling to inoculate in the culture medium;
(3) placing the inoculated kiwi fruit tissue culture seedling in a culture room with the culture temperature of 25 +/-1 ℃, the illumination time of 12 h/d and the illumination intensity of 1500-;
(4) the tissue culture seedling with delayed growth can be maintained for 4.5 months, and when the leaf of the tissue culture seedling is found to be dry, the new culture medium is replaced in time. The survival rate of the cultured tissue culture seedling reaches 83 percent.
The above description is only a preferred embodiment of the present invention, and all equivalent changes and modifications made in accordance with the claims of the present invention should be covered by the present invention.
Claims (1)
1. A culture method for prolonging the storage life of tissue culture seedlings of kiwi fruits is characterized in that: the method comprises the following steps:
(1) inoculating the non-pollution and strong-growing kiwi fruit tissue culture seedlings into a special subculture medium;
(2) storing the seeds in a culture chamber with the culture temperature of 25 +/-1 ℃, the illumination time of 12 h/d and the illumination intensity of 1500-;
(3) when the leaves of the tissue culture seedlings are dry, a new special subculture medium is replaced;
the special subculture medium is MS +6-BA 1.0 mg/L + NAA 1.0 mg/L + GA30.1 mg/L + agar 7g/L + sucrose 15 g/L.
Priority Applications (1)
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|---|---|---|---|
| CN201811603447.2A CN109430060B (en) | 2018-12-26 | 2018-12-26 | Culture method for prolonging storage life of kiwi fruit tissue culture seedlings |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201811603447.2A CN109430060B (en) | 2018-12-26 | 2018-12-26 | Culture method for prolonging storage life of kiwi fruit tissue culture seedlings |
Publications (2)
| Publication Number | Publication Date |
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| CN109430060A CN109430060A (en) | 2019-03-08 |
| CN109430060B true CN109430060B (en) | 2022-06-21 |
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| CN201811603447.2A Active CN109430060B (en) | 2018-12-26 | 2018-12-26 | Culture method for prolonging storage life of kiwi fruit tissue culture seedlings |
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Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112335548A (en) * | 2020-11-12 | 2021-02-09 | 福建省农业科学院果树研究所 | In-vitro conservation method of kiwi fruit germplasm resources |
Family Cites Families (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| USPP14212P3 (en) * | 2002-02-19 | 2003-10-07 | The Horticulture And Food Research Institute Of New Zealand Limited | Kiwi plant named ‘Hortgem Toru’ |
| CN101574056A (en) * | 2008-05-08 | 2009-11-11 | 黑龙江省林业科学研究所 | Actinidia arguta tissue culturing method |
| CN101647393B (en) * | 2009-09-24 | 2012-07-04 | 浙江省农业科学院 | Fast tissue culture reproducing method of actinidia eriantha |
| CN102177847B (en) * | 2011-03-07 | 2012-07-25 | 沈阳农业大学 | Factory seedling cultivating method of actinidia arguta |
| CN102845313A (en) * | 2012-10-19 | 2013-01-02 | 沈阳农业大学 | Method for quickly in-vitro actinidia kolomikta propagating |
| CN106258985B (en) * | 2016-09-30 | 2018-10-02 | 中国科学院华南植物园 | A kind of culture medium and cultural method extending tissue culture seedlings of bananas storage life |
| CN108739384A (en) * | 2018-05-31 | 2018-11-06 | 福建省农业科学院果树研究所 | A kind of method of Kiwifruit tissue-cultured seedling strengthening seedling and rooting |
| CN109006475B (en) * | 2018-07-16 | 2020-08-18 | 四川农业大学 | Kiwi fruit micro-grafting method and kiwi fruit seedling raising method |
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Application publication date: 20190308 Assignee: Sanming Dongfangliang Modern Agriculture Co.,Ltd. Assignor: POMOLOGY Research Institute FUJIAN ACADEMY OF AGRICULTURAL SCIENCES Contract record no.: X2024350000079 Denomination of invention: A cultivation method for extending the shelf life of kiwifruit tissue culture seedlings Granted publication date: 20220621 License type: Common License Record date: 20240514 |