CN107821167A - A kind of method for tissue culture of drinamyl potato - Google Patents
A kind of method for tissue culture of drinamyl potato Download PDFInfo
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- CN107821167A CN107821167A CN201711224056.5A CN201711224056A CN107821167A CN 107821167 A CN107821167 A CN 107821167A CN 201711224056 A CN201711224056 A CN 201711224056A CN 107821167 A CN107821167 A CN 107821167A
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- 235000002595 Solanum tuberosum Nutrition 0.000 title claims abstract description 45
- 244000061456 Solanum tuberosum Species 0.000 title claims abstract description 45
- 238000000034 method Methods 0.000 title claims abstract description 31
- 238000012360 testing method Methods 0.000 claims abstract description 15
- 230000001954 sterilising effect Effects 0.000 claims abstract description 12
- 239000007921 spray Substances 0.000 claims abstract description 10
- 238000001784 detoxification Methods 0.000 claims abstract description 9
- 230000008569 process Effects 0.000 claims abstract description 9
- 229960002523 mercuric chloride Drugs 0.000 claims abstract description 8
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 claims abstract description 8
- 239000002609 medium Substances 0.000 claims description 12
- 239000004576 sand Substances 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 241000196324 Embryophyta Species 0.000 claims description 6
- TWFZGCMQGLPBSX-UHFFFAOYSA-N Carbendazim Natural products C1=CC=C2NC(NC(=O)OC)=NC2=C1 TWFZGCMQGLPBSX-UHFFFAOYSA-N 0.000 claims description 5
- JNPZQRQPIHJYNM-UHFFFAOYSA-N carbendazim Chemical compound C1=C[CH]C2=NC(NC(=O)OC)=NC2=C1 JNPZQRQPIHJYNM-UHFFFAOYSA-N 0.000 claims description 5
- 239000006013 carbendazim Substances 0.000 claims description 5
- 238000005406 washing Methods 0.000 claims description 5
- 239000003643 water by type Substances 0.000 claims description 5
- 239000001963 growth medium Substances 0.000 claims description 4
- 238000007654 immersion Methods 0.000 claims description 2
- 230000004083 survival effect Effects 0.000 abstract description 8
- 241000700605 Viruses Species 0.000 abstract description 7
- 230000008030 elimination Effects 0.000 abstract description 7
- 238000003379 elimination reaction Methods 0.000 abstract description 7
- 238000004161 plant tissue culture Methods 0.000 abstract description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 7
- 238000005286 illumination Methods 0.000 description 6
- 230000006698 induction Effects 0.000 description 5
- 230000009286 beneficial effect Effects 0.000 description 4
- 230000004069 differentiation Effects 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 238000009395 breeding Methods 0.000 description 3
- 230000001488 breeding effect Effects 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 230000035755 proliferation Effects 0.000 description 3
- 230000003612 virological effect Effects 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 208000027418 Wounds and injury Diseases 0.000 description 2
- 229930014669 anthocyanidin Natural products 0.000 description 2
- 150000001452 anthocyanidin derivatives Chemical class 0.000 description 2
- 235000008758 anthocyanidins Nutrition 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 239000007943 implant Substances 0.000 description 2
- 208000014674 injury Diseases 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 239000013028 medium composition Substances 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- RMOGWMIKYWRTKW-UONOGXRCSA-N (S,S)-paclobutrazol Chemical compound C([C@@H]([C@@H](O)C(C)(C)C)N1N=CN=C1)C1=CC=C(Cl)C=C1 RMOGWMIKYWRTKW-UONOGXRCSA-N 0.000 description 1
- VEWUNRJQQRKSCB-UHFFFAOYSA-N 6-benzylpurine-2,6-diamine Chemical class C12=NC=NC2=NC(N)=NC1(N)CC1=CC=CC=C1 VEWUNRJQQRKSCB-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 229930191978 Gibberellin Natural products 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 239000005985 Paclobutrazol Substances 0.000 description 1
- 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 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 239000007640 basal medium Substances 0.000 description 1
- 230000003796 beauty Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- IXORZMNAPKEEDV-UHFFFAOYSA-N gibberellic acid GA3 Natural products OC(=O)C1C2(C3)CC(=C)C3(O)CCC2C2(C=CC3O)C1C3(C)C(=O)O2 IXORZMNAPKEEDV-UHFFFAOYSA-N 0.000 description 1
- 239000003448 gibberellin Substances 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 150000002475 indoles Chemical class 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000011785 micronutrient Substances 0.000 description 1
- 235000013369 micronutrients Nutrition 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 230000003405 preventing effect Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- UZKQTCBAMSWPJD-UQCOIBPSSA-N trans-Zeatin Natural products OCC(/C)=C\CNC1=NC=NC2=C1N=CN2 UZKQTCBAMSWPJD-UQCOIBPSSA-N 0.000 description 1
- UZKQTCBAMSWPJD-FARCUNLSSA-N trans-zeatin Chemical group OCC(/C)=C/CNC1=NC=NC2=C1N=CN2 UZKQTCBAMSWPJD-FARCUNLSSA-N 0.000 description 1
- 229940023877 zeatin Drugs 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
-
- 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
Abstract
The invention belongs to field of plant tissue culture technique, specifically discloses a kind of method for tissue culture of drinamyl potato.The method for tissue culture of the present invention comprises the following steps：(1) vernalization is handled；(2) choose explant and pre-process：The stem section of clip root tuber spray part is as explant；(3) sterilize：Soaked with mercuric chloride solution；(4) Initial culture：Explant after sterilizing is inoculated in Initial culture base and cultivated；(5) squamous subculture：After sterile bud is cut into stem section, it is inoculated in subculture multiplication medium and cultivates；(6) culture of rootage：Aseptic seedling is inoculated in root media and cultivated, obtains detoxification test tube plantlet.The method for tissue culture of the present invention carries out tissue cultures using the stem section of drinamyl potato children tender shoots as explant, sterile bud is induced using stem section culture, sterile bud survival rate and planting percent are high, detoxification test tube plantlet can be produced, test tube seedling is healthy and strong and virus elimination rate is high, effectively improves the quality of drinamyl potato.
Description
【Technical field】
The present invention relates to field of plant tissue culture technique, and in particular to a kind of method for tissue culture of drinamyl potato.
【Background technology】
Drinamyl potato nutritional value is high, and the content of contained anthocyanidin is up to 4.20mg/100g.Anthocyanidin has anti-
The various health-cares such as cancer, anti-aging, beauty and preventing hypertension, therefore, drinamyl potato have very high nutritive value and
Economic value, it can be used as Medical Nutritional product.
At present, the vegetative propagation of potato main tuberosity breeding, cutting propagation, plant division and mound layering and tissue training
Support, and the field production of potato is mainly bred using stem tuber now, stem tuber breeding has many advantages, but there is also obvious
The drawbacks of, i.e., potato seed dosage is big, breeding coefficient is low and viral easily accumulation, result in serious quality deterioration.
【The content of the invention】
The goal of the invention of the present invention is：For above-mentioned problem, there is provided a kind of tissue cultures of drinamyl potato
Method.The method for tissue culture of drinamyl potato of the present invention carries out group using the stem section of drinamyl potato children tender shoots as explant
Culture is knitted, realizes drinamyl potato stem section culture induction sterile bud, sterile bud survival rate and planting percent are high, can produce detoxification
Test tube seedling, test tube seedling is healthy and strong and virus elimination rate is high, beneficial to the healthy and strong growth for promoting drinamyl potato to be subsequently colonized, and effectively improves
The quality of drinamyl potato.
To achieve these goals, the technical solution adopted by the present invention is as follows：
A kind of method for tissue culture of drinamyl potato, comprises the following steps：
(1) vernalization is handled：Healthy drinamyl potato root tuber is chosen, is embedded in vernalization in sand bed, waters every other day, keep sand bed wet
Profit；
(2) choose explant and pre-process：Treat that the eye of drinamyl potato root tuber sprouts the height of seedling that grows of tender shoots to 10-
During 15cm, the downward 5-10cm in clip tender shoots top spray part, and cut off blade, leave 0.5cm petiole；Then, by institute
State spray and be cut into one axillary bud of band and a length of 1.5cm stem section；Mass concentration is used to soak institute for 1-5g/L carbendazim solution
Stem section 10-15min is stated, then 10-15min is rinsed with flowing water；
(3) sterilize：Pass through pretreated explant 4- with the mercuric chloride solution immersion that mass concentration is 0.1% is above-mentioned
After 7min, aseptic water washing 5 times；
(4) Initial culture：Explant after above-mentioned sterilizing is inoculated in Initial culture base and cultivated, obtains sterile bud；It is described
Initial culture base forms：1.8-2.3mg/L 6-BA, 0.2-0.5mg/L ZT and 0.2- are added in 1/2MS culture mediums
0.4mg/L GA3；
(5) squamous subculture：After above-mentioned sterile bud to be cut into the stem section of one axillary bud of band, it is inoculated in subculture multiplication medium
Culture, obtains aseptic seedling；The subculture multiplication medium forms：1.0-3.0mg/L 6-BA, 0.1- is added in MS culture mediums
0.6mg/L PP333With 0.4-0.8mg/L ZT；
(6) culture of rootage：Above-mentioned aseptic seedling is inoculated in root media by individual plant and cultivated, produces detoxicating cuvette
Seedling；The root media forms：In 1/2MS culture mediums add 0.1-0.6mg/L NAA, 1-1.5mg/L AC and
0.1-0.3mg/L IBA.
Further, being added dropwise in step (3), in the mercuric chloride solution has 3 drop Tween-80s.
Further, in step (4), the Initial culture base composition is：Add 2.0mg/L's in 1/2MS culture mediums
6-BA, 0.4mg/L ZT and 0.3mg/L GA3。
Further, in step (5), the length of the stem section is 1.2-1.5cm.
Further, in step (5), the subculture multiplication medium composition is：Add 2.5mg/L's in MS culture mediums
6-BA, 0.4mg/L PP333With 0.5mg/L ZT.
Further, in step (6), the root media composition is：Add 0.3mg/L's in 1/2MS culture mediums
NAA, 1.2mg/L AC and 0.2mg/L IBA.
Wherein, 1/2MS culture mediums are that the micronutrient levels in MS culture mediums is reduced into 1/2 obtained culture medium；6-BA
For 6- benzyl aminoadenines；ZT is zeatin；GA3For gibberellin；PP333For paclobutrazol；AC is activated carbon；IBA is indoles second
Acid.
In summary, by adopting the above-described technical solution, the beneficial effects of the invention are as follows：
(1) present invention is pre-processed and sterilization treatment to explant successively, pretreatment and sterilizing of the present invention
Processing combines, and effectively explant can be carried out disinfection, so as to effectively reduce the pollution rate of explant, and explant will not made
Into injury.
(2) culture of explant is carried out using the Initial culture base of the present invention, calli induction can be effectively facilitated simultaneously
Promote its differentiation and bud formation；Further, Multiplying culture is carried out to the stem section of sterile bud using the subculture multiplication medium of the present invention,
The survival rate and planting percent of stem section are high, can obtain the aseptic seedling of substantial amounts of stalwartness；Finally, using the present invention root media come
Culture of rootage is carried out to aseptic seedling, the effect of hestening rooting is good.
To sum up, the method for tissue culture of drinamyl potato of the present invention is used as explant by the use of the stem section of drinamyl potato children tender shoots
Body carries out tissue cultures, each step mutual cooperation effect, realizes drinamyl potato stem section culture induction sterile bud, sterile bud
Survival rate and planting percent are high, can produce detoxification test tube plantlet, test tube seedling is healthy and strong and virus elimination rate is high, after drinamyl potato is promoted
The healthy and strong growth of continuous field planting, effectively improve the quality of drinamyl potato.
【Embodiment】
Below in conjunction with specific embodiment, the present invention is further illustrated.
Embodiment 1
A kind of method for tissue culture of drinamyl potato, comprises the following steps：
(1) vernalization is handled：Healthy drinamyl potato root tuber is chosen, is embedded in vernalization in sand bed, waters every other day, keep sand bed wet
Profit；
(2) choose explant and pre-process：Treat that the eye of drinamyl potato root tuber sprouts the height of seedling that grows of tender shoots to 10cm
When, the downward 5cm in clip tender shoots top spray part, and cut off blade, leave 0.5cm petiole；Then, the spray is cut
Into the stem section of one axillary bud of band and a length of 1.5cm；Mass concentration is used to soak the stem section for 1g/L carbendazim solution
10min, then rinse 10min with flowing water；
(3) sterilize：The mercuric chloride solution that with mass concentration be 0.1% and dropwise addition has 3 drop Tween-80s soaks above-mentioned process
After pretreated explant 4min, aseptic water washing 5 times；
(4) Initial culture：Explant after above-mentioned sterilizing is inoculated in the Initial culture base that pH is 5.8 and cultivated, is cultivated
Temperature is 20 DEG C, daily illumination 12h, and intensity of illumination is 2000lx, obtains sterile bud；The Initial culture base forms：1/
1.8mg/L 6-BA, 0.2mg/L ZT and 0.2mg/L GA are added in 2MS culture mediums3；
(5) squamous subculture：After above-mentioned sterile bud to be cut into the stem section that length is one axillary bud of 1.2cm and band, pH is inoculated in
To be cultivated in 5.8 subculture multiplication medium, aseptic seedling is obtained；The subculture multiplication medium forms：Add in MS culture mediums
Add 1.0mg/L 6-BA, 0.1mg/L PP333With 0.4mg/L ZT；
(6) culture of rootage：Above-mentioned aseptic seedling is inoculated in the root media that pH is 5.7 by individual plant and cultivated, i.e.,
Obtain detoxification test tube plantlet；The root media forms：In 1/2MS culture mediums add 0.1mg/L NAA, 1mg/L AC and
0.1mg/L IBA.
Embodiment 2
A kind of method for tissue culture of drinamyl potato, comprises the following steps：
(1) vernalization is handled：Healthy drinamyl potato root tuber is chosen, is embedded in vernalization in sand bed, waters every other day, keep sand bed wet
Profit；
(2) choose explant and pre-process：Treat that the eye of drinamyl potato root tuber sprouts the height of seedling that grows of tender shoots to 13cm
When, the downward 8cm in clip tender shoots top spray part, and cut off blade, leave 0.5cm petiole；Then, the spray is cut
Into the stem section of one axillary bud of band and a length of 1.5cm；Mass concentration is used to soak the stem section for 3g/L carbendazim solution
12min, then rinse 13min with flowing water；
(3) sterilize：The mercuric chloride solution that with mass concentration be 0.1% and dropwise addition has 3 drop Tween-80s soaks above-mentioned process
After pretreated explant 6min, aseptic water washing 5 times；
(4) Initial culture：Explant after above-mentioned sterilizing is inoculated in the Initial culture base that pH is 5.8 and cultivated, is cultivated
Temperature is 22 DEG C, daily illumination 14h, and intensity of illumination is 2500lx, obtains sterile bud；The Initial culture base forms：1/
2.0mg/L 6-BA, 0.4mg/L ZT and 0.3mg/L GA are added in 2MS culture mediums3；
(5) squamous subculture：After above-mentioned sterile bud to be cut into the stem section that length is one axillary bud of 1.2-1.5cm and band, inoculation
Cultivated in the subculture multiplication medium that pH is 5.8, obtain aseptic seedling；The subculture multiplication medium forms：In MS culture mediums
Middle addition 2.5mg/L 6-BA, 0.4mg/L PP333With 0.5mg/L ZT；
(6) culture of rootage：Above-mentioned aseptic seedling is inoculated in the root media that pH is 5.7 by individual plant and cultivated, i.e.,
Obtain detoxification test tube plantlet；The root media forms：In 1/2MS culture mediums add 0.1mg/L NAA, 1mg/L AC and
0.1mg/L IBA.
Embodiment 3
A kind of method for tissue culture of drinamyl potato, comprises the following steps：
(1) vernalization is handled：Healthy drinamyl potato root tuber is chosen, is embedded in vernalization in sand bed, waters every other day, keep sand bed wet
Profit；
(2) choose explant and pre-process：Treat that the eye of drinamyl potato root tuber sprouts the height of seedling that grows of tender shoots to 15cm
When, the downward 10cm in clip tender shoots top spray part, and cut off blade, leave 0.5cm petiole；Then, by the spray
It is cut into one axillary bud of band and a length of 1.5cm stem section；Mass concentration is used to soak the stem section for 5g/L carbendazim solution
15min, then rinse 15min with flowing water；
(3) sterilize：The mercuric chloride solution that with mass concentration be 0.1% and dropwise addition has 3 drop Tween-80s soaks above-mentioned process
After pretreated explant 7min, aseptic water washing 5 times；
(4) Initial culture：Explant after above-mentioned sterilizing is inoculated in the Initial culture base that pH is 5.8 and cultivated, is cultivated
Temperature is 25 DEG C, daily illumination 16h, and intensity of illumination is 3000lx, obtains sterile bud；The Initial culture base forms：1/
2.3mg/L 6-BA, 0.5mg/L ZT and 0.4mg/L GA are added in 2MS culture mediums3；
(5) squamous subculture：After above-mentioned sterile bud to be cut into the stem section that length is one axillary bud of 1.5cm and band, pH is inoculated in
To be cultivated in 5.8 subculture multiplication medium, aseptic seedling is obtained；The subculture multiplication medium forms：Add in MS culture mediums
Add 3.0mg/L 6-BA, 0.6mg/L PP333With 0.8mg/L ZT；
(6) culture of rootage：Above-mentioned aseptic seedling is inoculated in the root media that pH is 5.7 by individual plant and cultivated, i.e.,
Obtain detoxification test tube plantlet；The root media forms：0.6mg/L NAA, 1.5mg/L AC are added in 1/2MS culture mediums
With 0.3mg/L IBA.
Compliance test result
1. pollution rate determines
To be pre-processed successively by the present invention with the explant of sterilization treatment as experimental group, with merely through pre- place of the invention
The explant of reason as a comparison case 1, with the explant merely through sterilization treatment of the present invention as a comparison case 2.By outside above-mentioned each group
Implant is inoculated on the basal medium without exogenous hormone (MS culture mediums+sucrose 18g/L+ agar 9g/L, pH=5.8) respectively
After carrying out culture 30d, there is situation about polluting (as long as having an explant in the blake bottle of inoculation explant in observation each group explant
There is the fungus colony being visually observed or bacterial clump and regards as polluting by whole bottle explant in body), and calculate outside each group
The pollution rate of implant, the results are shown in Table 1：
The pollution rate of each group explant of table 1
| Group | Experimental group | Comparative example 1 | Comparative example 2 |
| Pollution rate (%) | 1.1 | 8.4 | 3.2 |
As shown in Table 1, the pollution rate of the explant of experimental group 1 is significantly lower than comparative example 1 and 2, illustrate the present invention pretreatment and
Sterilization treatment combines, good to the Disinfection Effect of explant.
2. the observation of each cultivation stage of explant, statistical result
(1) bud rate is divided into the Initial cultures of above-mentioned 3 groups of embodiment explants and carries out observation statistics, the results are shown in Table 2：
The each group explant of table 2 is divided into bud rate
| Group | Embodiment 1 | Embodiment 2 | Embodiment 3 |
| It is divided into bud rate (%) | 97.3 | 98.6 | 96.7 |
As shown in Table 2, embodiment 1-3 explants are divided into that bud rate is higher, illustrate that the Initial culture base of the present invention can
Explant induction differentiation is effectively facilitated, simultaneously, it can be seen that and explant of the present invention pretreatment and sterilization steps are not made to explant
Into injury, the induction differentiation of explant is not influenceed.
(2) surviving for shoot proliferation culture is carried out to the sterile leaf stem section that above-described embodiment 1-3 obtains by Initial culture
Rate and planting percent carry out observation statistics, the results are shown in Table 3：
The survival rate and planting percent of the sterile leaf stem section of each group of table 3
| Group | Embodiment 1 | Embodiment 2 | Embodiment 3 |
| Survival rate (%) | 94.1 | 95.4 | 93.5 |
| Planting percent (%) | 91.1 | 92.8 | 90.3 |
As shown in Table 3, the sterile leaf stem section of each group is respectively provided with high survival rate and planting percent, illustrates the shoot proliferation of the present invention
Culture medium can effectively facilitate the healthy growth and differentiation of sterile bud.
(3) taking root for 7d culture of rootage is carried out to the aseptic seedling that above-described embodiment 1-3 obtains by shoot proliferation culture
Situation carries out observation statistics, the results are shown in Table 4：
The situation of taking root of each group aseptic seedling culture of rootage of table 4
| Group | Embodiment 1 | Embodiment 2 | Embodiment 3 |
| Rooting rate (%) | 100 | 100 | 100 |
| Take root and count (root) | 6.2 | 6.4 | 6.0 |
| Root shape condition | It is more sturdy | It is more sturdy | It is more sturdy |
As shown in Table 4, the rooting rate of each group aseptic seedling is 100%, and number of taking root is more and sturdy, illustrates taking root for the present invention
Culture medium is to the facilitation effect that has had of taking root of aseptic seedling, so as to beneficial to the survival rate improved when drinamyl potato is subsequently colonized
And the speed of growth.
(4) Viral diagnosis is carried out to the detoxification test tube plantlet that each group embodiment in (3) obtains, calculates virus elimination rate, the results are shown in Table
5：
The virus elimination rate of each group test tube seedling of table 5
| Group | Embodiment 1 | Embodiment 2 | Embodiment 3 |
| Virus elimination rate (%) | 89.5 | 92.3 | 90.4 |
As shown in Table 5, the test tube seedling virus elimination rate that the method for tissue culture by the present invention obtains is high, and viral level is few, from
And beneficial to promoting the healthy and strong of drinamyl potato to grow up, effectively improve the quality of drinamyl potato.
Described above is the detailed description for the present invention preferably possible embodiments, but embodiment is not limited to this hair
Bright patent claim, the equal change completed or modification change under the technical spirit suggested by all present invention, all should belong to
Cover the scope of the claims in the present invention.
Claims (6)
1. a kind of method for tissue culture of drinamyl potato, it is characterised in that comprise the following steps：
(1) vernalization is handled：Healthy drinamyl potato root tuber is chosen, is embedded in vernalization in sand bed, waters every other day, keep sand bed moistening；
(2) choose explant and pre-process：Treat that the eye of drinamyl potato root tuber sprouts the height of seedling that grows of tender shoots to 10-15cm
When, the downward 5-10cm in clip tender shoots top spray part, and cut off blade, leave 0.5cm petiole；Then, will be described tender
Stem section of the secateurs into one axillary bud of band and a length of 1.5cm；Mass concentration is used to soak the stem for 1-5g/L carbendazim solution
Section 10-15min, then rinse 10-15min with flowing water；
(3) sterilize：Pass through pretreated explant 4-7min with the mercuric chloride solution immersion that mass concentration is 0.1% is above-mentioned
Afterwards, aseptic water washing 5 times；
(4) Initial culture：Explant after above-mentioned sterilizing is inoculated in Initial culture base and cultivated, obtains sterile bud；The primary
Culture medium forms：1.8-2.3mg/L 6-BA, 0.2-0.5mg/L ZT and 0.2-0.4mg/ are added in 1/2MS culture mediums
L GA3；
(5) squamous subculture：After above-mentioned sterile bud to be cut into the stem section of one axillary bud of band, it is inoculated in subculture multiplication medium and trains
Support, obtain aseptic seedling；The subculture multiplication medium forms：1.0-3.0mg/L 6-BA, 0.1- is added in MS culture mediums
0.6mg/L PP333With 0.4-0.8mg/L ZT；
(6) culture of rootage：Above-mentioned aseptic seedling is inoculated in root media by individual plant and cultivated, produces detoxification test tube plantlet；
The root media forms：0.1-0.6mg/L NAA, 1-1.5mg/L AC and 0.1- are added in 1/2MS culture mediums
0.3mg/L IBA.
2. a kind of method for tissue culture of drinamyl potato according to claim 1, it is characterised in that described in step (3)
Being added dropwise in mercuric chloride solution has 3 drop Tween-80s.
3. a kind of method for tissue culture of drinamyl potato according to claim 1, it is characterised in that described in step (4)
Initial culture base forms：2.0mg/L 6-BA, 0.4mg/L ZT and 0.3mg/L GA are added in 1/2MS culture mediums3。
4. a kind of method for tissue culture of drinamyl potato according to claim 1, it is characterised in that described in step (5)
The length of stem section is 1.2-1.5cm.
5. a kind of method for tissue culture of drinamyl potato according to claim 1, it is characterised in that described in step (5)
Subculture multiplication medium forms：2.5mg/L 6-BA, 0.4mg/L PP are added in MS culture mediums333With 0.5mg/L's
ZT。
6. a kind of method for tissue culture of drinamyl potato according to claim 1, it is characterised in that described in step (6)
Root media forms：0.3mg/L NAA, 1.2mg/L AC and 0.2mg/L IBA are added in 1/2MS culture mediums.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN116491417A (en) * | 2023-03-29 | 2023-07-28 | 云南师范大学 | Regeneration method of potato wild species S.commersonii |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105248279A (en) * | 2015-10-28 | 2016-01-20 | 天津大学 | Tissue culture method for miniature potatoes |
| CN107047299A (en) * | 2017-03-13 | 2017-08-18 | 甘肃省农业科学院马铃薯研究所 | A kind of potato stem section tissue culture medium (TCM) and its cultural method |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105248279A (en) * | 2015-10-28 | 2016-01-20 | 天津大学 | Tissue culture method for miniature potatoes |
| CN107047299A (en) * | 2017-03-13 | 2017-08-18 | 甘肃省农业科学院马铃薯研究所 | A kind of potato stem section tissue culture medium (TCM) and its cultural method |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116491417A (en) * | 2023-03-29 | 2023-07-28 | 云南师范大学 | Regeneration method of potato wild species S.commersonii |
| CN116491417B (en) * | 2023-03-29 | 2024-04-16 | 云南师范大学 | Regeneration method of potato wild species S.commersonii |
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