CN108496798A - A kind of tissue culture propagation method of alum root " kimonos " - Google Patents
A kind of tissue culture propagation method of alum root " kimonos " Download PDFInfo
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- CN108496798A CN108496798A CN201810243389.0A CN201810243389A CN108496798A CN 108496798 A CN108496798 A CN 108496798A CN 201810243389 A CN201810243389 A CN 201810243389A CN 108496798 A CN108496798 A CN 108496798A
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- kimonos
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- alum root
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- 241000220152 Heuchera Species 0.000 title claims abstract description 43
- 238000000034 method Methods 0.000 title claims abstract description 28
- 230000001939 inductive effect Effects 0.000 claims abstract description 13
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 6
- 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 description 18
- 229930006000 Sucrose Natural products 0.000 claims description 18
- 229960004793 sucrose Drugs 0.000 claims description 18
- 229920001817 Agar Polymers 0.000 claims description 17
- 239000008272 agar Substances 0.000 claims description 17
- 239000006160 differential media Substances 0.000 claims description 17
- 238000005286 illumination Methods 0.000 claims description 13
- 239000005720 sucrose Substances 0.000 claims description 13
- 206010020649 Hyperkeratosis Diseases 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 230000006698 induction Effects 0.000 claims description 6
- 230000001954 sterilising effect Effects 0.000 claims description 6
- 235000013681 dietary sucrose Nutrition 0.000 claims description 5
- 230000004069 differentiation Effects 0.000 claims description 5
- 201000010099 disease Diseases 0.000 claims description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 4
- 238000005728 strengthening Methods 0.000 claims description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 3
- RCTYPNKXASFOBE-UHFFFAOYSA-M chloromercury Chemical compound [Hg]Cl RCTYPNKXASFOBE-UHFFFAOYSA-M 0.000 claims description 3
- 230000000249 desinfective effect Effects 0.000 claims description 3
- 239000008223 sterile water Substances 0.000 claims description 3
- 239000008399 tap water Substances 0.000 claims description 3
- 235000020679 tap water Nutrition 0.000 claims description 3
- 238000005406 washing Methods 0.000 claims description 3
- 238000011010 flushing procedure Methods 0.000 claims description 2
- 239000010977 jade Substances 0.000 claims 1
- 238000009395 breeding Methods 0.000 abstract description 6
- 230000001488 breeding effect Effects 0.000 abstract description 6
- 238000012258 culturing Methods 0.000 abstract description 4
- 230000008774 maternal effect Effects 0.000 abstract description 2
- 238000009331 sowing Methods 0.000 abstract description 2
- 241001529246 Platymiscium Species 0.000 abstract 1
- 230000035784 germination Effects 0.000 abstract 1
- 230000004083 survival effect Effects 0.000 abstract 1
- 239000001963 growth medium Substances 0.000 description 6
- 241000196324 Embryophyta Species 0.000 description 2
- 244000025254 Cannabis sativa Species 0.000 description 1
- 241000409198 Packera aurea Species 0.000 description 1
- 229940037003 alum Drugs 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000009403 interspecific hybridization Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000012883 rooting culture medium Substances 0.000 description 1
- 239000002352 surface water Substances 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
- A01H4/008—Methods for regeneration to complete plants
Landscapes
- 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 kind of tissue culture propagation methods of alum root " kimonos ", using the blade of alum root as explant, pass through explant disinfection, inducing clumping bud, subculture and strong sprout, outside sprout-cultivating-bottle series of steps, efficiently solve the problems, such as that new varieties alum root " kimonos " seed sowing germination percentage is low, maternal excellent leaf color and character are saved, achievees the purpose that produce fast-propagation.Tissue culture and rapid propagation method provided by the invention is suitable for the tissue-culturing rapid propagation of Saxifragaceae alum root " kimonos ", platymiscium biotechnology.Growth coefficient is 8 10 in 1 month, and rooting rate 95%, transplanting survival rate is 95% 98%, greatly improves the breeding coefficient of alum root " kimonos ", occupies for the kind rapid expansion market, provide effective propagation method.
Description
Technical field
The invention belongs to biotechnologies, and in particular, to a kind of tissue culture propagation of plant especially relates to
And a kind of important ornamental plant alum root " kimonos " tissue culture propagation method.
Background technology
Alum root " kimonos " belongs to the new varieties that Saxifragaceae alum root category alum root interspecific hybridization obtains, perennial cold-resistant draft flower
Grass, shallow root, resistance to -34 DEG C of low temperature have higher ornamental value.It is chiefly used in hayashishita flower border, ground quilt, courtyard greening etc. in gardens
Deng.The kind of alum root is more, some kind tillering abilities are strong, and then tillering ability is weak for some.It, can be numerous by sowing in actual production
The alum root type grown is relatively limited, a kind of culture medium of tissue-culturing quick-propagations of alum root blade of CN201710118920.7 and
It with blade is method that explant carries out tissue culture propagation to be proposed in cultural method a kind of, because kind is numerous in alum root, each kind
Feature difference is larger, therefore is not fully appropriate for the fast numerous of all alum roots, has no to be directed at present and is somebody's turn to do the use of " kimonos " new varieties
Blade carries out the report of the method for tissue culture propagation.
Invention content：
Alum root " kimonos " is the kind of selection and breeding in alum root Interspecific Hybrids, has excellent ornamental value strain, seminal propagation cannot
The consistency for ensureing offspring, is generally bred by tissue culture method, the present invention is intended to provide a kind of alum root " kimonos " utilizes leaf
Piece tissue culture propagation method accelerates the reproduction speed of alum root, more seedlings is provided for market.
In order to realize the above-mentioned purpose of the present invention, the present invention provides the following technical solutions：
A kind of tissue culture propagation method of alum root " kimonos ", this method include the following steps：
(1) callus induction：The blade that will be obtained after sterilization process is cut into the small of 1cm*1cm with sterile curved scissors
Block is seeded in inducing culture, and light culture enters alternation of light and darkness culture 12h/12h after 3-5 days, is cultivated 30-40 days, illumination is strong
Degree is 2000-3000LUX, and the inducing culture is MS+1mg/L KT+0.2mg/L TDZ+1mg/L IBA, sucrose 28g/
L, agar 7g/L, pH6.0；
(2) callus that will be induced, is seeded in differential medium, be placed in the brightness time be 10h/8h environment under,
Intensity of illumination is 2000-3000LUX, is cultivated 20-30 days, and the differential medium is MS+1mg/L 6-BA+0.5mg/LKT+
0.1mg/L IBA, sucrose 28g/L, agar 7g/L, pH6.0；
(3) strong sprout and culture of rootage：The sprout that 1.5cm~2cm high is grown in differential medium is forwarded to strong sprout life
Root culture medium is additional saccharose 30g/L, agar 7g/L, pH6.0 in MS+0.3mg/L IBA+0.5mg/LNAA culture mediums；Strong sprout
And take root and be placed in luminous intensity to be cultivated 20-30 days under 1500LUX-2500LUX, the brightness time is 12h/10h.
A kind of tissue culture propagation method of alum root " kimonos ", this method comprises the following steps：
1) explant selects：The disease-free blade for taking alum root " kimonos ", circulating water rinses 10h-12h standby under tap water
With；
2) explant sterilizes：It takes the blade got ready on sterile super-clean bench, with 75% alcohol disinfecting 20s, uses sterile water
3-5 is rinsed after, 9min is sterilized with 0.1% mercury chloride, using aseptic water washing 3-5 all over spare；
3) callus induction：Inducing culture is MS+1mg/L KT+0.2mg/L TDZ+1mg/L IBA, sucrose 28g/
L, agar 7g/L, pH6.0.The blade that will be obtained after sterilization process blots surface water on super-clean bench with aseptic filter paper
Point, with sterile curved scissors, it is cut into the fritter of 1cm*1cm, is seeded in inducing culture, light culture enters alternation of light and darkness after 3-5 days
12h/12h is cultivated, is cultivated 30-40 days, intensity of illumination 2000-3000LUX；
4) differentiation culture：Differential medium be MS+1mg/L 6-BA+0.5mg/LKT+0.1mg/L IBA, sucrose 28g/L,
Agar 7g/L, pH6.0；The callus that will be induced is seeded in differential medium (Fig. 2), and it is 10h/ to be placed in the brightness time
It is cultivated 20-30 days under 8h environment, temperature is 20-23 DEG C, intensity of illumination 2000-3000LUX；
5) strong sprout and culture of rootage：The sprout that 1.5cm~2cm high is grown in differential medium is forwarded to strengthening seedling and rooting
Culture medium is additional saccharose 30g/L, agar 7g/L, pH6.0 in MS+0.3mg/L IBA+0.5mg/LNAA (Fig. 2, Fig. 3)；
It is cultivate 20-30 days under 1500LUX-2500LUX that strong sprout and taking root, which is placed in luminous intensity, and temperature is 20-23 DEG C, and the brightness time is
12h/10h。
Compared with prior art, the beneficial effects of the invention are as follows：
1. innovating and realizing the tissue culture propagation method of new varieties alum root " kimonos "；
2. the prior art is bred using explant terminal bud or axillary bud, and the present invention breaches and uses the kind leaf
Piece is the method that explant carries out tissue culture propagation；
3. by tissue culture expanding propagation, alum root " kimonos " growth coefficient within half a year is 20-30 times, greatly improve alum root " and
The breeding coefficient of clothes ", a kind of culture mediums of the tissue-culturing quick-propagation of alum root blade of significantly larger than CN201710118920.7
And the breeding potential of cultural method.
Description of the drawings：
Fig. 1 is that alum root " kimonos " breaks up seedling figure；
Fig. 2 is that alum root " kimonos " is taken root figure；
Fig. 3 is that alum root " kimonos " is taken root figure.
Specific implementation mode：
Below in conjunction with the accompanying drawings, the essentiality content further illustrated the present invention with the embodiment of the present invention, but not with
This limits the present invention.
Embodiment 1：
Alum root " kimonos " tissue cultures, carry out as follows：
1. the preparation of culture medium：Configure inducing culture MS culture medium MS+1mg/L KT+0.2mg/L TDZ+1mg/L
IBA, sucrose 28g/L, agar 7g/L, pH6.0；Configuration differentiation culture：MS+1mg/L6-BA+0.5mg/LKT+0.1mg/L IBA,
Sucrose 28g/L, agar 7g/L, pH6.0；Configure strong sprout and root media：MS+0.3mg/L IBA+0.5mg/LNAA, sucrose
30g/L, agar 7g/L adjust pH6.0；
2, concrete operation step：
(1) explant is acquired：Alum root " kimonos " is one of existing alum root kind more popular on the market, what the present invention used
Alum root " kimonos " is bought from Yunnan Province's academy of agricultural science, and disease-free blade is won from alum root " kimonos " maternal plant；
(2) callus induction：The blade that will be obtained after sterilization process is blotted on super-clean bench with aseptic filter paper
Surface moisture is cut into the fritter of 1cm*1cm, is seeded in inducing culture with sterile curved scissors, alternation of light and darkness 12h/12h, culture
30-40 days, intensity of illumination 2000-3000LUX；
(3) differentiation culture：The callus that will be induced, is seeded in differential medium, and first light culture is placed in after 5 days
The brightness time is to be cultivated 20-30 days under 10h/8h environment, and temperature is 20-23 DEG C, intensity of illumination 2000-3000LUX；
(4) strong sprout and culture of rootage：The small sprout switching strengthening seedling and rooting culture of 3 leaves will be grown in differential medium
In base, it is to be cultivated 20-30 days under 1500LUX-2500LUX to be placed in luminous intensity, and temperature is 20-23 DEG C, and the brightness time is 12h/
10h。
Embodiment 2
The alum offspring for carrying out tissue culture will be needed to be placed in a greenhouse culture 1 month, daily clear water pours, and takes alum root " kimonos "
Disease-free blade, it is spare after circulating water flushing 12h under tap water；
1) explant sterilizes：It takes the blade got ready on sterile super-clean bench, with 75% alcohol disinfecting 20s, uses sterile water
3-5 is rinsed after, 9min is sterilized with 0.1% mercury chloride, using aseptic water washing 3-5 all over spare；
2) callus induction：Inducing culture is MS+1mg/L KT+0.2mg/L TDZ+1mg/L IBA, sucrose 28g/
L, agar 7g/L, pH6.0, light culture carry out illumination cultivation 30-40 days after 5 days, the brightness time is 12h/12h.Sterilizing will be passed through
The terminal bud obtained after process blots surface moisture on super-clean bench with aseptic filter paper, with sterile curved scissors, is cut into the small of 1cm*1cm
Block is seeded in inducing culture, is cultivated 30-40 days, intensity of illumination 2000-3000LUX；
3) differentiation culture：Differential medium be MS+1mg/L 6-BA+0.5mg/LKT+0.1mg/L IBA, sucrose 28g/L,
Agar 7g/L, pH6.0；The callus that will be induced, is seeded in differential medium, and first light culture is after 5 days, when being placed in brightness
Between under 12h/8h environment, intensity of illumination 2000-3000LUX；
4) strong sprout and culture of rootage：It is MS+ that the small sprout that 3 leaves are grown in differential medium, which is forwarded to culture medium,
In the culture medium of 0.3mg/L IBA+0.5mg/LNAA, additional saccharose 30g/L, agar 7g/L, pH6.0；Strong sprout and taking root is placed in
Luminous intensity is to be cultivated 20-30 days under 1500LUX-2500LUX, and temperature is 20-23 DEG C, and the brightness time is 12h/10h.
By tissue culture expanding propagation, alum root " kimonos " of the present invention growth coefficient within half a year is 20-30 times, greatly improves alum root
A kind of culture of the tissue-culturing quick-propagation of alum root blade of the breeding coefficient of " kimonos ", significantly larger than CN201710118920.7
The breeding potential of base and cultural method.
Claims (2)
1. a kind of tissue culture propagation method of alum root " kimonos ", it is characterised in that this method includes the following steps：
(1) callus induction：Alum root " kimonos " blade obtained after sterilization process is cut into 1cm* with sterile curved scissors
The fritter of 1cm, is seeded in inducing culture, and light culture enters alternation of light and darkness culture 12h/12h after 3-5 days, cultivates 30-40
It, intensity of illumination 2000-3000LUX, the inducing culture is MS+1mg/L KT+0.2mg/L TDZ+1mg/L
IBA, sucrose 28g/L, agar 7g/L, pH6.0；
(2) callus that will be induced, is seeded in differential medium, and it is illumination under 10h/8h environment to be placed in the brightness time
Intensity is 2000-3000LUX, is cultivated 20-30 days, and the differential medium is MS+1mg/L 6-BA+0.5mg/LKT+
0.1mg/L IBA, sucrose 28g/L, agar 7g/L, pH6.0；
(3) strong sprout and culture of rootage：The sprout that 1.5cm-2cm high is grown in differential medium is forwarded to strengthening seedling and rooting culture
Base is additional saccharose 30g/L, agar 7g/L, pH6.0 in MS+0.3mg/L IBA+0.5mg/LNAA culture mediums；It strong sprout and takes root
It is to be cultivated 20-30 days under 1500LUX-2500LUX to be placed in luminous intensity, and the brightness time is 12h/10h.
2. a kind of tissue culture propagation method of alum root " kimonos ", it is characterised in that this method comprises the following steps：
1) explant selects：The disease-free blade for taking alum root " kimonos ", it is spare after circulating water flushing 10h-12h under tap water；
2) explant sterilizes：It takes the blade 1) got ready on sterile super-clean bench, with 75% alcohol disinfecting 20s, is rushed with sterile water
3-5 is washed after, 9min is sterilized with 0.1% mercury chloride, it is spare using aseptic water washing 3-5 times；
3) callus induction：Inducing culture is MS+1mg/L KT+0.2mg/L TDZ+1mg/L IBA, sucrose 28g/L, fine jade
Fat 7g/L, pH6.0, the blade that will be obtained after sterilization process blot surface moisture on super-clean bench with aseptic filter paper, use
Sterile curved scissors is cut into the fritter of 1cm*1cm, is seeded in inducing culture, and light culture enters alternation of light and darkness culture after 3-5 days
12h/12h is cultivated 30-40 days, intensity of illumination 2000-3000LUX；
4) differentiation culture：Differential medium is MS+1mg/L 6-BA+0.5mg/LKT+0.1mg/L IBA, sucrose 28g/L, agar
7g/L, pH6.0, the callus that will be induced, are seeded in differential medium, and it is to be trained under 10h/8h environment to be placed in the brightness time
It supports 20-30 days, temperature is 20-23 DEG C, intensity of illumination 2000-3000LUX；
5) strong sprout and culture of rootage：The sprout that 1.5cm-2cm high is grown in differential medium is forwarded to strengthening seedling and rooting culture
Base is additional saccharose 30g/L, agar 7g/L, pH6.0 in MS+0.3mg/L IBA+0.5mg/LNAA；Strong sprout and taking root is placed in
Luminous intensity is to be cultivated 20-30 days under 1500LUX-2500LUX, and temperature is 20-23 DEG C, and the brightness time is 12h/10h.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201810243389.0A CN108496798A (en) | 2018-03-23 | 2018-03-23 | A kind of tissue culture propagation method of alum root " kimonos " |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810243389.0A CN108496798A (en) | 2018-03-23 | 2018-03-23 | A kind of tissue culture propagation method of alum root " kimonos " |
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| Publication Number | Publication Date |
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| CN108496798A true CN108496798A (en) | 2018-09-07 |
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| CN201810243389.0A Pending CN108496798A (en) | 2018-03-23 | 2018-03-23 | A kind of tissue culture propagation method of alum root " kimonos " |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109430056A (en) * | 2018-11-19 | 2019-03-08 | 上海市农业科学院 | A method of induction alum root adventitious shoot regeneration |
| CN117598199A (en) * | 2024-01-24 | 2024-02-27 | 云南农业大学 | A kind of alum root hybrid breeding method |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106942051A (en) * | 2017-03-02 | 2017-07-14 | 浙江省萧山棉麻研究所 | The culture medium and propagation method of a kind of tissue-culturing quick-propagation of alum root blade |
-
2018
- 2018-03-23 CN CN201810243389.0A patent/CN108496798A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106942051A (en) * | 2017-03-02 | 2017-07-14 | 浙江省萧山棉麻研究所 | The culture medium and propagation method of a kind of tissue-culturing quick-propagation of alum root blade |
Non-Patent Citations (2)
| Title |
|---|
| 李小东等: "矾根组培快繁", 《中国花卉园艺》 * |
| 陈锦等: "美洲矾根愈伤组织诱导及其快繁技术体系研究", 《湖北农业科学》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109430056A (en) * | 2018-11-19 | 2019-03-08 | 上海市农业科学院 | A method of induction alum root adventitious shoot regeneration |
| CN109430056B (en) * | 2018-11-19 | 2021-11-09 | 上海市农业科学院 | Method for inducing regeneration of adventitious buds of alum roots |
| CN117598199A (en) * | 2024-01-24 | 2024-02-27 | 云南农业大学 | A kind of alum root hybrid breeding method |
| CN117598199B (en) * | 2024-01-24 | 2024-03-22 | 云南农业大学 | A kind of alum root hybrid breeding method |
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Application publication date: 20180907 |