CN112189561A - Application of sodium hydrosulfide as plant callus inducer - Google Patents
Application of sodium hydrosulfide as plant callus inducer Download PDFInfo
- Publication number
- CN112189561A CN112189561A CN202011055892.7A CN202011055892A CN112189561A CN 112189561 A CN112189561 A CN 112189561A CN 202011055892 A CN202011055892 A CN 202011055892A CN 112189561 A CN112189561 A CN 112189561A
- Authority
- CN
- China
- Prior art keywords
- sodium hydrosulfide
- callus
- plant
- inducer
- nahs
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Developmental Biology & Embryology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses an application of sodium hydrosulfide as a plant callus inducer, which is characterized in that the sodium hydrosulfide is prepared into an aqueous solution with the concentration of 20-80 mu M, the aqueous solution is placed in a sterilized 200 mu L centrifugal tube, and the centrifugal tube is placed in a corresponding culture medium, so that the induction rate of callus can be improved, the formation of plant callus is promoted, and the regeneration frequency of adventitious buds is improved. The invention provides a new direction for applying sodium hydrosulfide, namely the sodium hydrosulfide is used for promoting the formation of plant callus and improving the differentiation rate of adventitious buds in the process of plant tissue culture.
Description
Technical Field
The invention belongs to the technical field of plant tissue culture, and particularly relates to application of sodium hydrosulfide as a plant callus inducer.
Background
Plant tissue culture has facilitated genetic, physiological, biochemical and pathological studies in plants and has become a routine method for plant science research. The tissue culture of the plant is beneficial to the fine variety breeding of the plant and the preservation of fine germplasm resources, and is an important basis for obtaining high-yield and high-quality varieties through a genetic engineering technology. The callus means a tissue which is newly generated on the surface of a wound after a part of an original plant body is stimulated by a wound, and the tissue is composed of living parenchyma cells and can be originated from living cells of various tissues in any organ of the plant body. In plant tissue culture, callus formation and morphogenesis are critical steps. The induction of the callus is a precondition for establishing plant tissue culture, and the formation of the rapidly induced callus can greatly shorten the time for plant tissue culture and improve the efficiency of plant tissue culture.
Hydrogen sulfide (H)2S) is a third gas signal molecule following Nitric Oxide (NO) and carbon monoxide (CO). Sodium hydrosulfide (NaHS) is a commonly used hydrogen sulfide (H)2S) donor, dissolving NaHS in water to release H2S gas, the principle of which is the hydrosulfide radical ion (HS) of NaHS-) With hydrogen ions (H) in water+) Bound to release H2S, the specific reaction formula is as follows: HS-+H+＝H2And S. At present H2S has been studied mainly on H2S relieving action and mechanism of action during biotic and abiotic stress. H2S participates in a plurality of physiological metabolic processes, such as seed germination, root morphogenesis, photosynthesis, leaf senescence delay, plant stress resistance improvement and the like. However, there has been no report on the effect of hydrogen sulfide on the induction of callus formation during plant tissue culture.
Disclosure of Invention
The invention aims to provide an application of NaHS as a plant callus inducer.
In order to achieve the purpose, the invention adopts the following technical scheme:
the invention provides an application of sodium hydrosulfide as a plant callus inducer, and the NaHS is applied to plant tissue culture as the inducer.
Further, the NaHS is used as an inducer to promote plant callus formation during plant tissue culture.
Further, the NaHS is used as an inducer to increase the adventitious bud differentiation rate during plant tissue culture.
Furthermore, the NaHS is prepared into NaHS aqueous solution with the concentration of 20-80 mu M, when in use, the NaHS aqueous solution is prepared as before, and plant tissues formed by the callus under induction are fumigated, so that the callus inductivity and the callus weight gain factor can be obviously improved, and the adventitious bud regeneration frequency is improved.
According to the invention, NaHS is prepared into a NaHS aqueous solution with a certain concentration, Chinese characteristic vegetable Chinese cabbage is taken as an experimental material, the stem cotyledon of the Chinese cabbage is fumigated, and the experimental result shows that compared with the condition without the NaHS, the wound part of the stem cotyledon treated by the NaHS fumigation is obviously expanded, and the capability of callus for inducing adventitious buds is obviously enhanced. The method shows that NaHS has the effect of promoting callus formation, and can shorten the tissue culture period and improve the efficiency of plant tissue culture in the process of plant tissue culture.
The NaHS aqueous solution of the invention is prepared by slowly releasing H in water2S, it plays the role of promoting the formation of callus.
Compared with the prior art, the invention has the following beneficial effects:
1. the NaHS is used as an inducer, has the effect of promoting the formation of the callus, and can improve the induction rate of the callus;
2. the NaHS is used as an inducer, so that the weight gain times of the callus are increased, and the effect of shortening the culture period of plant tissues is achieved;
3. the NaHS adopted by the invention has the advantages of good water solubility, low cost, safety, no toxicity, simple operation and obvious effect.
Description of the drawings:
FIG. 1 shows callus induction conditions after treatment with NaHS aqueous solutions of different concentrations;
FIG. 2 is a comparison of callus induction rates after treatment with different concentrations of NaHS aqueous solution;
FIG. 3 is a comparison of callus weight gain after treatment with NaHS aqueous solutions of different concentrations;
FIG. 4 shows the differentiation of adventitious buds after treatment with NaHS aqueous solutions of different concentrations;
FIG. 5 is a graph showing the comparison of the adventitious bud differentiation rates after treatment with different concentrations of NaHS aqueous solutions.
Detailed Description
The present invention will be described in further detail with reference to specific examples. The embodiment is implemented on the premise of the technical solution of the present invention, and a detailed implementation manner and a specific operation process are given, but the protection scope of the present invention is not limited to the embodiment.
The embodiment of the invention provides application of sodium hydrosulfide as an inducer in plant tissue culture, wherein the plant tissue is from a Chinese cabbage cotyledon with stalk.
The sodium hydrosulfide disclosed by the embodiment of the invention is applied to the caulicle leaves of the Chinese cabbage as an inducer, so that the formation of callus of the caulicle leaves of the Chinese cabbage is promoted, and the differentiation rate of adventitious buds is improved.
The NaHS provided by the embodiment of the invention is prepared into a NaHS aqueous solution with the concentration of 20-80 mu M, and is prepared when in use.
The embodiment of the invention provides a using method for rapidly inducing and forming callus of stem-bearing cotyledon of Chinese cabbage, which specifically comprises the following steps:
(1) selecting full and uniform sized Chinese cabbage seed (Beijing New No. three), sterilizing with 75% anhydrous ethanol for 30S, and 0.1% HgCl2Sterilizing for 6min, washing with sterilized water for 5 times, uniformly sowing in 1/2MS culture medium, placing in illumination incubator for 5 days, and culturing under the following conditions: the culture temperature is 23 deg.C, relative humidity is 60%, and illumination intensity is 160 μ Emm-2s-1The light/dark was 16/8 h.
(2) Cutting stem leaves of the Chinese cabbage seedlings in the step (1) at the growing point to be used as explants, inserting the stem leaves into 4 bottles of identical induced callus culture media, respectively placing 25 stem leaves on each bottle of culture media, wherein the induced culture media contain 2 mg/L6-BA and 0.3mg/L NAA, and adding 7mg/L AgNO into the culture media3The browning is prevented; putting NaHS aqueous solutions with different concentrations (0, 20, 50 and 80 mu M) into a sterilized 200 mu L centrifugal tube, and putting the centrifugal tube into 4 bottles of culture medium respectively to fumigate the stemmed leaves, wherein the fumigation culture conditions are as follows: the culture temperature is 23 deg.C, relative humidity is 60%, and illumination intensity is 160 μ Emm-2s-1The light/dark was 16/8 h.
(3) After 5 days of treatment, the callus expansion degree (figure 1) and the induction rate differentiation rate (figure 2) are observed, and the callus weight gain times (figure 3) are counted, wherein the formula is as follows:
as shown in FIG. 1, the wounded part of cotyledon callus with petiole treated with NaHS was significantly enlarged compared with that without NaHS; as shown in FIG. 2, the same letter indicates that the statistical result is on the same level, different letters indicate that the result is not on the same level, the callus induction rate has significant difference, and the callus induction rate is obviously improved after the NaHS treatment; as shown in figure 3, the same letter indicates that the statistical result is on the same level, different letters indicate that the result is not on the same level, the callus weight gain times have significant difference, the callus weight gain times are obviously improved after the NaHS is added for treatment, and the culture period is shortened.
(4) After 7 days of treatment, the degree of adventitious bud differentiation was observed (FIG. 4), and the rate of adventitious bud differentiation was counted (FIG. 5), and the formula was as follows:
as shown in FIG. 4, the cotyledons with stalks treated with NaHS showed significantly more adventitious shoots than those treated with NaHS; as shown in FIG. 5, the same letter indicates that the statistical results are on the same level, different letters indicate that the results are not on the same level, the adventitious bud differentiation rate has significant difference, and the adventitious bud differentiation rate is obviously improved after NaHS treatment.
Claims (7)
1. Application of sodium hydrosulfide as plant callus inducer.
2. The use of sodium hydrosulfide as an inducer of plant callus according to claim 1, characterized in that: the sodium hydrosulfide is used as an inducer for plant tissue culture.
3. The use of sodium hydrosulfide as an inducer of plant callus according to claim 2, characterized in that: the sodium hydrosulfide is used as an inducer for promoting the formation of plant callus in the process of plant tissue culture.
4. The use of sodium hydrosulfide as an inducer of plant callus according to claim 2, characterized in that: the sodium hydrosulfide is used as an inducer for improving the differentiation rate of adventitious buds in the process of plant tissue culture.
5. The method for using sodium hydrosulfide as plant callus induction agent according to any one of claims 1 to 4, characterized in that: the sodium hydrosulfide is prepared into a sodium hydrosulfide aqueous solution with the concentration of 5-500 mu M, and the sodium hydrosulfide aqueous solution is used for fumigating induction culture of plant tissues formed by the callus under induction.
6. The method for using sodium hydrosulfide as plant callus induction agent according to any one of claims 1 to 4, characterized in that: the sodium hydrosulfide is prepared into a sodium hydrosulfide aqueous solution with the concentration of 20-80 mu M, and the sodium hydrosulfide aqueous solution is used for fumigating induction culture of plant tissues formed by the callus under induction.
7. The method for using sodium hydrosulfide as plant callus inducer according to claim 6, characterized by that: the fumigation induction culture conditions are as follows: the culture temperature is 23 deg.C, relative humidity is 60%, and illumination intensity is 160 μ Emm-2s-1The light/dark was 16/8 h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011055892.7A CN112189561A (en) | 2020-09-29 | 2020-09-29 | Application of sodium hydrosulfide as plant callus inducer |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011055892.7A CN112189561A (en) | 2020-09-29 | 2020-09-29 | Application of sodium hydrosulfide as plant callus inducer |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112189561A true CN112189561A (en) | 2021-01-08 |
Family
ID=74008229
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011055892.7A Pending CN112189561A (en) | 2020-09-29 | 2020-09-29 | Application of sodium hydrosulfide as plant callus inducer |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112189561A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040072790A1 (en) * | 2002-10-09 | 2004-04-15 | Yaguang Liu | Safe natural pharmaceutical composition for treating cancer |
| CN101385465A (en) * | 2008-10-15 | 2009-03-18 | 合肥工业大学 | New use of sodium hydrosulfide for promoting plant root morphogenesis |
| CN102851347A (en) * | 2012-07-13 | 2013-01-02 | 东北林业大学 | New application of hydrogen sulfide in promotion of secondary metabolite synthesis |
| CN107223668A (en) * | 2017-05-12 | 2017-10-03 | 郑龙 | A kind of plant graft consolidant |
| CN111528101A (en) * | 2020-06-18 | 2020-08-14 | 陈滋倩 | Differentiation culture medium for carnation anther culture |
-
2020
- 2020-09-29 CN CN202011055892.7A patent/CN112189561A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040072790A1 (en) * | 2002-10-09 | 2004-04-15 | Yaguang Liu | Safe natural pharmaceutical composition for treating cancer |
| CN101385465A (en) * | 2008-10-15 | 2009-03-18 | 合肥工业大学 | New use of sodium hydrosulfide for promoting plant root morphogenesis |
| CN102851347A (en) * | 2012-07-13 | 2013-01-02 | 东北林业大学 | New application of hydrogen sulfide in promotion of secondary metabolite synthesis |
| CN107223668A (en) * | 2017-05-12 | 2017-10-03 | 郑龙 | A kind of plant graft consolidant |
| CN111528101A (en) * | 2020-06-18 | 2020-08-14 | 陈滋倩 | Differentiation culture medium for carnation anther culture |
Non-Patent Citations (2)
| Title |
|---|
| ZHONG-GUANG LI,JIA-ZHENG JIN: "Hydrogen sulfide partly mediates abscisic acid-induced heat tolerance in tobacco (Nicotiana tabacum L.) suspension cultured cells", 《PLANT CELL, TISSUE AND ORGAN CULTURE 》 * |
| 戴莹等: "药用植物组织培养中褐化现象的研究进展", 《中草药》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Deepika et al. | In vitro regeneration of Punica granatum L. plants from different juvenile explants | |
| CN105191793B (en) | The efficient breeding method of purple tissue culture seedlings of potatoes | |
| Do et al. | Effects of nano silver on the growth of banana (Musa spp.) cultured in vitro | |
| Ahmad et al. | Optimization of protocol for surface sterilization and callus induction for three rice varieties | |
| CN113317204B (en) | Method for inducing adventitious buds of seedlings of sedum aizoon and efficiently regenerating plants | |
| CN110810247A (en) | Sweet potato stem tip detoxification and breeding method | |
| CN109392721A (en) | A method of induction sweet potato plant regeneration | |
| CN117617122A (en) | Combined culture medium for small Huang Jiangtuo toxin and thin-layer culture and culture method | |
| CN111567178A (en) | Breeding method for improving germination rate of August fried seeds | |
| CN116686717A (en) | Method for breeding small Huang Jiangjing tip detoxified stock | |
| CN112189561A (en) | Application of sodium hydrosulfide as plant callus inducer | |
| CN108401906B (en) | Method for grafting pomelo by using micro-buds | |
| CN116019013A (en) | Fast breeding method of bougainvillea spectabilis | |
| CN113854155B (en) | High-throughput breeding method of vanilla virus-free seedlings | |
| Roh et al. | The influence of silver thiosulfate and thidiazuron on shoot regeneration from cotyledon explants of Brassica napus | |
| CN110089430B (en) | Eucommia ulmoides seedling culture method | |
| CN108293879B (en) | Rapid propagation method of euphorbia kansui | |
| CN106134999A (en) | The method that capital potato 6 group training detoxification is cultivated | |
| Khan et al. | EFFECT OF DIFFERENT HORMONAL CONCENTRATIONS ONIN-VITRO REGENERAT-ION OF RICE (Oryza sativaL.) | |
| Budisantoso et al. | Effect of BAP (6-Benzyl Amino Purine) Concentration on Growth Micro Cutting of Nepenthes ampullaria | |
| CN112042535B (en) | Induction method of kok-saghyz tetraploid plant | |
| CN116649215B (en) | Tissue culture and rapid propagation method of phyllanthus niruri | |
| Karn et al. | In-vitro regeneration in long-day garlic (Allium sativum) | |
| CN111972074B (en) | Seed treatment method for early-maturing sweet cherry variety and method for sowing seedlings in current year | |
| CN115956506B (en) | Construction method of tea tree in-vitro regeneration tissue |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210108 |
|
| RJ01 | Rejection of invention patent application after publication |