CN109329054A - A kind of vitrified method of reduction mulberries adventitious bud - Google Patents
A kind of vitrified method of reduction mulberries adventitious bud Download PDFInfo
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- CN109329054A CN109329054A CN201811215287.4A CN201811215287A CN109329054A CN 109329054 A CN109329054 A CN 109329054A CN 201811215287 A CN201811215287 A CN 201811215287A CN 109329054 A CN109329054 A CN 109329054A
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- Prior art keywords
- mulberries
- adventitious bud
- bud
- vitrified
- reduction
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- 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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
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- 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 belongs to plant tissue culture fast breeding technique fields, disclose a kind of vitrified method of reduction mulberries adventitious bud, and glass rate gradually rises with the increase of cultivation temperature, and at 22 DEG C, mulberries adventitious bud grows best, proliferation rate highest；Moisture content (81.87%) soluble sugar content (14.37mg/g) is extremely significant to be lower than other Temperature Treatments, PAL activity (43.08U/ (gmin)), chlorophyll content (1.48mg/g), proliferation rate (1.59) is extremely significant to be higher than other Temperature Treatments, is the optimum culturing temperature for preventing the vitrified generation of mulberries adventitious bud.Temperature is vitrified to mulberries adventitious bud to be had a significant impact；The present invention provides certain reference frame for the control and large-scale production of glass effect in mulberries tissue culture procedures.
Description
Technical field
The invention belongs to plant tissue culture fast breeding technique field more particularly to a kind of vitrified sides of reduction mulberries adventitious bud
Method.
Background technique
Currently, the prior art commonly used in the trade is such that
Mulberries (Fructus Mori) are perennial xylophytas, nickname sorosis, mulberries, after being the solid development of mulberry tree
A kind of aggregate fruit, amino acid necessary to fruit protein rich in, various human bodies and a variety of be easy to be inhaled by human body
The fructose and glucose of receipts, moreover also containing the multivitamins such as VB1-VB6, VE, VC, the mineral such as Fe, Zn, Ca member
The substance very useful to human body such as element, cellulose and carrotene has anti-oxidation stress, anti-radiation, antitumor, elimination inflammation
Disease improves cardiovascular function and prevents the effect of the health cares such as coronary heart disease.
Mulberries are mainly bred using grafting and cuttage, and breeding coefficient is low, it is difficult to which satisfaction commercially produces demand, group
It is largely to provide the effective way of seedlings, but tissue culture discovery adventitious bud vitrifying is extremely serious, studies and controls glass that training is numerous fastly
The generation of change promotes the sound development of mulberries group culturation rapid propagating technology to be of great significance.
Vitrifying refers to the fast numerous some translucent lopsided test tube plants formed in the process of plant tissue culture, referred to as " glass
Glass seedling ", in vitro culture, the phenomenon that regeneration plant grows into " vitreous shoot ", is known as " vitrification phenomenon ".Vitrifying seedling proliferation
Rate is low, it is also difficult to take root.
The short and small swelling of vitrifying plant, blade, tender tip are translucent shape；Stem is short and thick, almost without internode；Blade is thick and narrow
Long, shrinkage or curling, it is fragile easily broken；Leaf table anoderm, non-functional stomatal apparatus, leaf only have spongy tissue, without fence
Tissue.The dry matter of vitreous shoot, chlorophyll content, protein content decline, cellulose and content of lignin are low, cuticula, grid
Column tissue development is not complete, and performance of taking root is very poor, hydroxycinnamic acid related with lignin synthesis, COA ligase and and lignifying
The activity of the related phenylalanine ammonia lyase of process is more much lower than field run plant.
In conclusion problem of the existing technology is:
(1) mulberries sapling multiplication is generally based on cuttage, propagation by grafiting, it is difficult to which satisfaction commercially produces demand.
(2) group culturation rapid propagating technology, adventitious bud proliferation rate is higher, and remains kind merit, but adventitious bud vitrifying
It is abnormal serious.
(3) vitrifying adventitious bud proliferation rate is low, it is difficult to take root, it is difficult to commercial applications.
Solve the meaning of above-mentioned technical problem:
Vitrification generation is related with many factors, and wherein temperature is main affecting factors, and research control mulberries are not
The vitrified temperature condition of normal bud reduces vitrifying proportion, and the sound development of mulberries group culturation rapid propagating technology is promoted to have weight
Want meaning.
Summary of the invention
In view of the problems of the existing technology, the present invention provides a kind of vitrified methods of reduction mulberries adventitious bud.
The invention is realized in this way a kind of vitrified method of reduction mulberries adventitious bud includes:
Under the conditions of growth cabinet, cultivated at a temperature of 18 DEG C, 22 DEG C, 26 DEG C, 30 DEG C, 34 DEG C respectively, illumination is
12h/d。
It selects and is cultivated at a temperature of 22 DEG C, illumination 12h/d.For best practice.
Further, the vitrified method of the reduction mulberries adventitious bud further comprises:
Mulberries Adventitious bud culture is carried out using culture medium；Culture medium are as follows: MS+6-BA 1.0mg/L+NAA 0.01mg/L+
+ 0.6% agar of 3% sucrose.
Further, the reduction vitrified method of mulberries adventitious bud further comprises: the calculating of proliferation rate, specifically:
Total bud number/initial bud number after proliferation rate=proliferation.
Further, the vitrified method of the reduction mulberries adventitious bud further comprises: phenylalanine ammonia lyase is active
It calculates, specifically:
Phenylalanine ammonia lyase activity=(sample cell reaction solution OD value-inactivation control tube reaction solution OD value) × sample extraction
Liquid total volume/(samples taken extracting liquid volume time of enzymatic reacting sample quality when 0.01 × measurement).
In conclusion advantages of the present invention and good effect are shown in Table 1:
The influence that mulberries adventitious bud vitrifying occurs for 1 temperature of table
It can be seen that in the present invention: temperature is vitrified to mulberries adventitious bud to be had a significant impact, and glass rate is with culture
The increase of temperature and gradually rise, at 22 DEG C, mulberries adventitious bud grow best, proliferation rate highest.Moisture content (81.87%),
Soluble sugar content (14.37mg/g) is extremely significant to be lower than other Temperature Treatments, PAL activity (43.08U/ (g
Min)), chlorophyll content (1.48mg/g), it is to prevent mulberries indefinite that proliferation rate (1.59) is extremely significant, which to be higher than other Temperature Treatments,
The optimum culturing temperature of the vitrified generation of bud.For the control and large-scale production of glass effect in mulberries tissue culture procedures
Certain reference frame is provided.
Detailed description of the invention
Fig. 1 is the vitrified method flow diagram of reduction mulberries adventitious bud provided in an embodiment of the present invention.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to
Limit the present invention.
It is low that the prior art not can effectively solve vitrifying seedling proliferation rate, it is difficult to the problem of taking root.
Fig. 1 is the vitrified method of reduction mulberries adventitious bud provided in an embodiment of the present invention, comprising:
S101: it under the conditions of growth cabinet, is cultivated at a temperature of 18 DEG C, 22 DEG C, 26 DEG C, 30 DEG C, 34 DEG C respectively, illumination
It is 12h/d.It selects and is cultivated at a temperature of 22 DEG C, illumination 12h/d.For best practice.
Wherein, mulberries Adventitious bud culture is carried out using culture medium；Culture medium are as follows: MS+6-BA 1.0mg/L+NAA0.01mg/
+ 0.6% agar of L+3% sucrose.
S102: the calculating of proliferation rate, specifically: total bud number/initial bud number after proliferation rate=proliferation.Phenylalanine ammonolysis
The calculating of enzymatic activity, specifically: phenylalanine ammonia lyase activity=(sample cell reaction solution OD value-inactivation control tube reaction solution OD
Value) × sample extracting solution total volume/(0.01 × samples taken extracting liquid volume time of enzymatic reacting sample quality when measuring.
Below with reference to concrete analysis, the invention will be further described.
In the vitrified method of reduction mulberries adventitious bud provided in an embodiment of the present invention,
Culture medium prescription :+0.6% agar of MS+6-BA 1.0mg/L+NAA 0.01mg/L+3% sucrose.
Condition of culture: temperature (18,22,26,30,34) DEG C, illumination 12h/d.
Proliferation rate calculates: total bud number/initial bud number after proliferation rate=proliferation.
Phenylalanine ammonia lyase activity=(sample cell reaction solution OD value-inactivation control tube reaction solution OD value) × sample extraction
Liquid total volume/(samples taken extracting liquid volume time of enzymatic reacting sample quality when 0.01 × measurement).
In the present invention: temperature is vitrified to mulberries adventitious bud to be had a significant impact, and glass rate is with cultivation temperature
Increase and gradually rise, at 22 DEG C, mulberries adventitious bud grow best, proliferation rate highest.Moisture content (81.87%) is soluble
Sugared content (14.37mg/g) it is extremely significant be lower than other Temperature Treatments, PAL activity (43.08U/ (gmin)),
Chlorophyll content (1.48mg/g), it is to prevent mulberries adventitious bud glass that proliferation rate (1.59) is extremely significant, which to be higher than other Temperature Treatments,
The optimum culturing temperature of the generation of change.One is provided for the control and large-scale production of glass effect in mulberries tissue culture procedures
Fixed reference frame.
Below with reference to specific experiment, the invention will be further described.
Mulberries adventitious bud under 18 degrees Celsius are as follows: adventitious bud green, slow growth, seedling are short and small.
Mulberries adventitious bud under 22 degrees Celsius are as follows: adventitious bud green, growth is very fast, and seedling is higher, and bud is more.
Mulberries adventitious bud under 26 degrees Celsius are as follows: adventitious bud light green color, growth is very fast, and seedling is higher, and bud is more.
Mulberries adventitious bud under 30 degrees Celsius are as follows: adventitious bud is light yellow, and growth is slower, and seedling is shorter, and bud is less.
Mulberries adventitious bud under 34 degrees Celsius are as follows: adventitious bud blade yellow, growth is slower, and seedling is shorter, and bud is few.
Mulberries adventitious bud under 18-34 degrees Celsius of different temperatures are as follows: it is increased with temperature, adventitious bud increment gradually decreases,
Leaf color gradually becomes yellow from green.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Made any modifications, equivalent replacements, and improvements etc., should all be included in the protection scope of the present invention within mind and principle.
Claims (4)
1. a kind of vitrified method of reduction mulberries adventitious bud, which is characterized in that the vitrified side of the reduction mulberries adventitious bud
Method includes:
Under the conditions of growth cabinet, cultivated at a temperature of 18 DEG C, 22 DEG C, 26 DEG C, 30 DEG C, 34 DEG C respectively, illumination is 12h/
d。
2. reducing the vitrified method of mulberries adventitious bud as described in claim 1, which is characterized in that the reduction mulberries are indefinite
The vitrified method of bud further comprises:
Mulberries Adventitious bud culture is carried out using culture medium；Culture medium are as follows: MS+6-BA 1.0mg/L+NAA 0.01mg/L+3% sugarcane
Sugared+0.6% agar.
3. reducing the vitrified method of mulberries adventitious bud as described in claim 1, which is characterized in that the reduction mulberries are indefinite
The vitrified method of bud further comprises: the calculating of proliferation rate, specifically:
Total bud number/initial bud number after proliferation rate=proliferation.
4. reducing the vitrified method of mulberries adventitious bud as described in claim 1, which is characterized in that the reduction mulberries are indefinite
The vitrified method of bud further comprises: the active calculating of phenylalanine ammonia lyase, specifically:
Phenylalanine ammonia lyase activity=(sample cell reaction solution OD value-inactivation control tube reaction solution OD value) × sample extracting solution is total
Volume/(samples taken extracting liquid volume time of enzymatic reacting sample quality when 0.01 × measurement).
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| CN201811215287.4A CN109329054A (en) | 2018-10-18 | 2018-10-18 | A kind of vitrified method of reduction mulberries adventitious bud |
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| CN201811215287.4A CN109329054A (en) | 2018-10-18 | 2018-10-18 | A kind of vitrified method of reduction mulberries adventitious bud |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109197592A (en) * | 2018-10-24 | 2019-01-15 | 河南云帮农业科技有限公司 | A method of reducing albumen mulberry tissue culture Vitrification |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103120128A (en) * | 2013-02-28 | 2013-05-29 | 河南科技大学 | Method for avoiding paeonia suffruticosa tissue culture seedling browning and vitrifying |
| CN104026010A (en) * | 2014-06-04 | 2014-09-10 | 江苏科技大学 | Inducing method for adventitious buds of mulberry cotyledons |
-
2018
- 2018-10-18 CN CN201811215287.4A patent/CN109329054A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103120128A (en) * | 2013-02-28 | 2013-05-29 | 河南科技大学 | Method for avoiding paeonia suffruticosa tissue culture seedling browning and vitrifying |
| CN104026010A (en) * | 2014-06-04 | 2014-09-10 | 江苏科技大学 | Inducing method for adventitious buds of mulberry cotyledons |
Non-Patent Citations (2)
| Title |
|---|
| 李娅莉等: "观赏植物组织培养过程中玻璃化现象与解决措施进展", 《四川农业大学学报》 * |
| 王振龙主编: "《植物组织培养》", 30 June 2007, 中国农业大学出版社 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109197592A (en) * | 2018-10-24 | 2019-01-15 | 河南云帮农业科技有限公司 | A method of reducing albumen mulberry tissue culture Vitrification |
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Application publication date: 20190215 |