CN106718896B - A kind of subculture method of Bulgaria essential oil rose - Google Patents
A kind of subculture method of Bulgaria essential oil rose Download PDFInfo
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- CN106718896B CN106718896B CN201611119737.0A CN201611119737A CN106718896B CN 106718896 B CN106718896 B CN 106718896B CN 201611119737 A CN201611119737 A CN 201611119737A CN 106718896 B CN106718896 B CN 106718896B
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- 241000220317 Rosa Species 0.000 title claims abstract description 69
- 239000000341 volatile oil Substances 0.000 title claims abstract description 23
- 238000000034 method Methods 0.000 title claims abstract description 10
- 241000196324 Embryophyta Species 0.000 claims abstract description 22
- 239000000463 material Substances 0.000 claims abstract description 18
- 238000002474 experimental method Methods 0.000 claims abstract description 8
- 239000012879 subculture medium Substances 0.000 claims abstract description 6
- 241001582888 Lobus Species 0.000 claims abstract description 5
- 238000011081 inoculation Methods 0.000 claims abstract description 5
- 235000000659 Rosa rugosa Nutrition 0.000 claims description 9
- 240000006066 Rosa rugosa Species 0.000 claims description 9
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 claims description 9
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 6
- 239000008103 glucose Substances 0.000 claims description 6
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 4
- 241000206575 Chondrus crispus Species 0.000 claims description 4
- 229910021580 Cobalt(II) chloride Inorganic materials 0.000 claims description 4
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 claims description 4
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 claims description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 4
- 239000007836 KH2PO4 Substances 0.000 claims description 4
- 229910004619 Na2MoO4 Inorganic materials 0.000 claims description 4
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 claims description 4
- 239000001110 calcium chloride Substances 0.000 claims description 4
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 4
- 229910052927 chalcanthite Inorganic materials 0.000 claims description 4
- 229910052564 epsomite Inorganic materials 0.000 claims description 4
- 238000005286 illumination Methods 0.000 claims description 4
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 claims description 4
- 229960000367 inositol Drugs 0.000 claims description 4
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 claims description 4
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 4
- 229910052603 melanterite Inorganic materials 0.000 claims description 4
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 4
- 235000001968 nicotinic acid Nutrition 0.000 claims description 4
- 229960003512 nicotinic acid Drugs 0.000 claims description 4
- 239000011664 nicotinic acid Substances 0.000 claims description 4
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 claims description 4
- 239000011684 sodium molybdate Substances 0.000 claims description 4
- TVXXNOYZHKPKGW-UHFFFAOYSA-N sodium molybdate (anhydrous) Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 claims description 4
- 229960000344 thiamine hydrochloride Drugs 0.000 claims description 4
- 235000019190 thiamine hydrochloride Nutrition 0.000 claims description 4
- 239000011747 thiamine hydrochloride Substances 0.000 claims description 4
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 claims description 4
- 229910000368 zinc sulfate Inorganic materials 0.000 claims description 4
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims description 4
- 239000011686 zinc sulphate Substances 0.000 claims description 4
- 238000004519 manufacturing process Methods 0.000 abstract description 7
- 206010023126 Jaundice Diseases 0.000 abstract description 3
- 230000034303 cell budding Effects 0.000 abstract description 3
- 239000004615 ingredient Substances 0.000 abstract description 3
- 206010053759 Growth retardation Diseases 0.000 abstract description 2
- 231100000001 growth retardation Toxicity 0.000 abstract description 2
- 238000011534 incubation Methods 0.000 abstract description 2
- 235000016709 nutrition Nutrition 0.000 abstract description 2
- 238000011160 research Methods 0.000 description 11
- 239000005556 hormone Substances 0.000 description 6
- 229940088597 hormone Drugs 0.000 description 6
- 229920001817 Agar Polymers 0.000 description 5
- 239000008272 agar Substances 0.000 description 5
- 238000009395 breeding Methods 0.000 description 5
- 230000001488 breeding effect Effects 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 235000011449 Rosa Nutrition 0.000 description 4
- 229930006000 Sucrose Natural products 0.000 description 4
- 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 4
- 238000012136 culture method Methods 0.000 description 4
- 239000005720 sucrose Substances 0.000 description 4
- 238000012090 tissue culture technique Methods 0.000 description 4
- 238000012549 training Methods 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 244000020518 Carthamus tinctorius Species 0.000 description 2
- 235000004789 Rosa xanthina Nutrition 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 159000000007 calcium salts Chemical class 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000003205 fragrance Substances 0.000 description 2
- 235000021552 granulated sugar Nutrition 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 239000003921 oil Substances 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 239000005631 2,4-Dichlorophenoxyacetic acid Substances 0.000 description 1
- 235000003255 Carthamus tinctorius Nutrition 0.000 description 1
- 206010054949 Metaplasia Diseases 0.000 description 1
- 244000181025 Rosa gallica Species 0.000 description 1
- 235000000533 Rosa gallica Nutrition 0.000 description 1
- 241000109329 Rosa xanthina Species 0.000 description 1
- 241000220222 Rosaceae Species 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 239000000039 congener Substances 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000015689 metaplastic ossification Effects 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 230000001902 propagating effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 235000000346 sugar Nutrition 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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
-
- 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)
- Cultivation Of Plants (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
A kind of subculture method of Bulgaria essential oil rose, comprising the following steps: (1) experimental material must select；(2) specific experiment step: the acquisition of (2.1) aseptic explant: excision blade retains terminal bud lobus cardiacus, and petiole stays 0.1-0.2cm, is cut into the stem section of 0.5-0.8cm；(2.3) Bulgarian essential oil rose squamous subculture；Terminal bud and stem section separate inoculation, are equably inoculated into subculture medium 1 (MG-1)；(2.4) after cultivating 35d, plant longitudinal growth plant height is sprouted in 3.5cm or more, plant lateral bud, and lower part has a lateral budding is long to form single size about 2cm, and statistics multiplied ratio averagely reaches 1:4.5.Nutritional ingredient needed for the present invention is directed to rose squamous subculture incubation is adjusted, and solves the phenomenon that plant jaundice growth retardation in rose tissue-cultured seedling Subculture；And improve the multiplied ratio of rose tissue-cultured seedling.It is successfully established the tissue culture system of rose, realizes rose the factorial production.
Description
Technical field
The present invention relates to a kind of tissue culture methods, and in particular to a kind of subculture method of Bulgaria essential oil rose belongs to
In field of biotechnology.
Background technique
Bulgarian Rosa rugosa category rosaceae, Rosa, machaka.It is fuel-displaced since Bulgarian Rosa rugosa belongs to international odor type
Rate is up to 3/10000ths. six, is the best kind extracted Rosa Damascana and produce rose (fresh flower water).Bulgarian rose
Rare yield is very big, can achieve 500~750 kilograms of per mu yield fresh flower, 80~100 kilograms of per mu yield dried flower, dried flower flower bud.
The rose of Bulgaria is universally acknowledged high-quality rose variety, and the fragrance of a flower is pure, careful, belongs to international light odor type, flower
Phase Relatively centralized gives out fresh and sweet fragrance when blooming, be to extract Rosa Damascana and process the best kind of rose flavored juice, thus quilt
Plantation has very high economic value to extract attar of rose extensively.It the rose industry of Bulgaria existing more than 300 years goes through
History is known as the good reputation of " rose kingdom ".The rose produced with Bulgarian Rosa rugosa fresh flower, per kilogram price are beautiful up to 1000
Member.The Rosa Damascana extracted from Bulgarian Rosa rugosa fresh flower, per kilogram price are up to 7000~8000 dollars, are known as " liquid
Gold " title.
Bulgarian Rosa rugosa is adaptable, and cultivation management is easy, high financial profit.To the north of Yangtze River in China and on the south Beijing
Qu Junke cultivation.It is the new industry with broad mass market prospect and huge exploitation potential.
Existing research technology:
In the existing report about rose tissue cultures, rose tissue cultures mostly are carried out by point of penetration of hormone combinations
Research.Deng Yanhong etc. is white using MS+6-BA2.0mg/L+NAA 0.01mg/L+3% in " rose Study on tissue culture " report
This formula of sugar+0.4g/L agar carries out the Multiplying culture of rose, and proliferation times can reach 6 or more；
Zhao Yipeng etc. also uses the similar formula of class in " tissue culture technique of rose " research, only to the concentration of hormone
It is adjusted, minimal medium selects MS, and the variation range of hormone is in BA1.0mg/L~10mg/L, GA3 0.25-20mg/
L, NAA and IAA 0.01-0.05mg/L；
Zhang Yanqiu etc. proposes that rose is in subculture medium MS+6-BA3.0mg/L in " rose tissue culture rapid propagating technology "
Growth coefficient highest, can achieve 4 in+NAA 0.1mg/L；
In keep it is superfine studied using purple branch rose as test material when " researchs of purple branch rose tissue cultures ", using MS
+ 1.8mg/l 6-BA+0.08mg/L NAA+0.4mg/l GA3 effect is fine；
" pre-test of white flower Rose plants tissue cultures " and " researchs of safflower Rose plants tissue cultures " are respectively with white flower rose
Rare and safflower rose uses identical formula MS+6-BA 3.0mg/L+NAA 0.2mg/ as test material, squamous subculture stage
L+ sucrose 30g/L+ agar 6g/L, multiplied ratio highest is respectively 3.24 and 3.75.
" edible rose tissue-culturing rapid propagation key technology research " and " Study on tissue culture of black red rose " selects Yunnan to work as
The edible rose kind " Mo Hong " on ground is used as test material, and the culture formula of multiplicative stage is respectively MS+6-BA3.0mg/L+NAA
0.1mg/L+ sucrose 30g/l+ agar 6g/l and MS+6-BA2.5mg/L+NAA 0.05mg/L+ sucrose 30g/l+ agar 6g/l increases
Growing coefficient is respectively 3.5 and 3.517.
It is basic training with MS using " Damascus " rose as material in " research of essential oil rose tissue culture technique " text
Support base, by Orthogonal Experiment and Design, obtained conclusion are as follows: hormone is handled with 6BA1.0mg/L and NAA0.01mg/l, culture
Carbon source selects white granulated sugar dosage best for cultivation effect of the 40g/L to rose in base.
The suitable Damask Rose Multiplying culture that " research of Damask Rose tissue culture technique " is obtained through overtesting
Formula be WPM+6-BA3.0mg/L+NAA 0.1mg/L+ sucrose 30g/l+ agar 6g/l, growth coefficient can reach 5.1.
Breeding " Bulgaria " rose that " rose tissue cultures and fast breeding technique experimental study " are introduced with Gansu province into
Row group training research obtain aseptic explant and enter the Multiplying culture stage, using MS+BA1.0+2,4-D 0.1, MS+BA1.0+
This 3 kinds formulas of IBA0.1 and MS+BA1.0+NAA0.2 are used interchangeably, and the rose tissue-cultured seedling of high proliferation coefficient is obtained with this.
Shortcoming is reported in existing research:
The studies above report rose material used can be divided into ornamental rose, edible rose and smart rose for oil.Plant
The theoretical basis of tissue culture technique be cellular omnipotency theory, even and congener, because genotype difference, tissue cultures
Used in squamous subculture technology it is also variant.Aromatic substance is abundant compared with other 2 class roses in smart rose for oil body, therefore straight
Connect the demand influenced during its tissue culture for each nutriment and Auto-regulator.It is more in culture medium in above-mentioned report
It is cultivated with 6-BA (concentration is greater than 2.0) collocation NAA of high concentration, 6-BA mostly generation may be decreased tissue-cultured seedling using easily accumulation
Differentiation capability, while group training research are carried out by material of " Bulgaria " rose, it is carried out using 3 kinds of formula interactions, this is for work
The production of factory's metaplasia easily causes managerial confusion, is not easy to carry out the control of production technology.
The present invention can reach 4.5 or more using " Bulgaria " rose as test material, using a kind of formula growth coefficient, at
The factorial production of function realization rose.
Summary of the invention
The object of the present invention is to provide a kind of subculture methods of Bulgarian essential oil rose, to overcome existing skill
Disadvantages mentioned above present in art and deficiency.
This experiment solves a kind of Bulgarian essential oil rose for a kind of Subculture of Bulgarian essential oil rose
Multiplied ratio is low in rare tissue-culturing rapid propagation, tissue-cultured seedling jaundice and vitrified problem, to realize Bulgarian essential oil rose tissue culture
The factorial production.
Technical problems to be solved needed for the present invention can be achieved through the following technical solutions:
A kind of quick breeding by group culture method of Bulgaria essential oil rose, it is characterised in that: the following steps are included:
(1) experimental material must select: select growing way is neat, plant height 2.5-4.0cm, subculture cycle are 30-35 days,
Leaf color is in the Bulgarian Rosa rugosa tissue-cultured seedling of green or peak green as squamous subculture material；
(2) specific experiment step:
(2.1) cutting of material: excision blade retains terminal bud lobus cardiacus, and petiole stays 0.1-0.2cm, is cut into 0.5-0.8cm
Stem section；
(2.2) Bulgarian essential oil rose squamous subculture；Terminal bud and stem section separate inoculation, are equably inoculated into squamous subculture
In base 1 (MG-1)；
Wherein, MS (KNO is improved in step (2.2)31.9-2.5g/L, NH4NO31.65-2.1g/L MgSO4·7H2O
0.37-0.48g/L, KH2PO40.17-0.22g/L, CaCl2·2H2O 0.44-0.57g/L, Na2EDTA·2H2O 0.041-
0.056g/L,FeSO4·7H2O 0.03-0.041g/L,MnSO4·H2O 16.9mg/L, H3BO36.2mg/L, ZnSO4·7H2O
8.6mg/L, KI 0.83mg/L, Na2MoO4·2H2O0.25mg/L, CoCl2·6H2O 0.025mg/L, CuSO4·5H2O
0.025mg/L, inositol 10mg/L, niacin 0.05mg/L, thiamine hydrochloride 0.01mg/L, puridoxine hydrochloride 0.05mg/L)+6-
BA0.3mg/L+NAA 0.01mg/L+ glucose 20g/L+ carragheen 5.5g/L；
Wherein, in step (2.2), it be light application time is 12h/d that Bulgarian essential oil rose squamous subculture, which obtains condition of culture,
Intensity of illumination 2000-3000lux, cultivation temperature are 26 ± 2 DEG C；
(2.3) after cultivating 35d, plant longitudinal growth plant height is sprouted in 3.5cm or more, plant lateral bud, and lower part has one
Lateral budding is long to form single size about 2cm, and statistics multiplied ratio averagely reaches 1:4.5.
Beneficial effects of the present invention:
The present invention be directed to rose squamous subculture incubation needed for nutritional ingredient be adjusted, improve a great number of elements,
The dosage of molysite, calcium salt, carbon source use glucose instead, solve the phenomenon that plant jaundice growth retardation during rose tissue-cultured seedling；
Simultaneously using the combined treatment of the 6-BA and NAA of low concentration, reduce because hormone is excessively high and mostly generation accumulation leads to rose
There is abnormal risk in tissue-cultured seedling differentiation capability, and improves the multiplied ratio of rose tissue-cultured seedling.
This experiment improves rose minimal medium ingredient and using suitable hormone combinations, is matched using one group of Multiplying culture
Side is successfully established the tissue culture system of rose, realizes rose the factorial production.
Specific embodiment
Below in conjunction with specific embodiment, progress explanation is made to the present invention.It should be understood that following embodiment is merely to illustrate this hair
It is bright not for limiting the scope of the invention.
Embodiment 1
A kind of quick breeding by group culture method of Bulgaria essential oil rose, comprising the following steps:
(1) experimental material must select: select growing way is neat, plant height 2.5-4.0cm, subculture cycle are 30-35 days,
Leaf color is in the Bulgarian Rosa rugosa tissue-cultured seedling of green or peak green as squamous subculture material；
(2) specific experiment step:
(2.1) acquisition of aseptic explant: excision blade retains terminal bud lobus cardiacus, and petiole stays 0.1-0.2cm, is cut into
The stem section of 0.5-0.8cm；
(2.2) Bulgarian essential oil rose squamous subculture；Terminal bud and stem section separate inoculation, are equably inoculated into squamous subculture
In base 1 (MG-1)；
Wherein, MS (KNO is improved in step (2.2)31.9-2.5g/L, NH4NO31.65-2.1g/L MgSO4·7H2O
0.37-0.48g/L, KH2PO40.17-0.22g/L, CaCl2·2H2O 0.44-0.57g/L, Na2EDTA·2H2O 0.041-
0.056g/L,FeSO4·7H2O 0.03-0.041g/L,MnSO4·H2O 16.9mg/L, H3BO36.2mg/L, ZnSO4·7H2O
8.6mg/L, KI 0.83mg/L, Na2MoO4·2H2O0.25mg/L, CoCl2·6H2O 0.025mg/L, CuSO4·5H2O
0.025mg/L, inositol 10mg/L, niacin 0.05mg/L, thiamine hydrochloride 0.01mg/L, puridoxine hydrochloride 0.05mg/L)+6-
BA0.3mg/L+NAA 0.01mg/L+ glucose 20g/L+ carragheen 5.5g/L；
Wherein, in step (2.2), it be light application time is 12h/d that Bulgarian essential oil rose squamous subculture, which obtains condition of culture,
Intensity of illumination 2000-3000lux, cultivation temperature are 26 ± 2 DEG C；
As described in Table 1, pH is adjusted to 5.9 for growth performance in each subculture medium:
Table 1
Table 1 the results showed that compared with white granulated sugar, be added in the culture medium of glucose, rose tissue-cultured seedling growth
Well, degree of lignification is low, is not in early ageing phenomenon, and plant is full of vitality.
Embodiment 2
A kind of quick breeding by group culture method of Bulgaria essential oil rose, comprising the following steps:
(1) experimental material must select: select growing way is neat, plant height 2.5-4.0cm, subculture cycle are 30-35 days,
Leaf color is in the Bulgarian Rosa rugosa tissue-cultured seedling of green or peak green as squamous subculture material；
(2) specific experiment step:
(2.1) acquisition of aseptic explant: excision blade retains terminal bud lobus cardiacus, and petiole stays 0.1-0.2cm, is cut into
The stem section of 0.5-0.8cm；
(2.2) Bulgarian essential oil rose squamous subculture；Terminal bud and stem section separate inoculation, are equably inoculated into squamous subculture
In base 1 (MG-1)；
Wherein, MS (KNO is improved in step (2.3)31.9-2.5g/L, NH4NO31.65-2.1g/L MgSO4·7H2O
0.37-0.48g/L, KH2PO40.17-0.22g/L, CaCl2·2H2O 0.44-0.57g/L, Na2EDTA·2H2O 0.041-
0.056g/L,FeSO4·7H2O 0.03-0.041g/L,MnSO4·H2O 16.9mg/L, H3BO36.2mg/L, ZnSO4·7H2O
8.6mg/L, KI 0.83mg/L, Na2MoO4·2H2O0.25mg/L, CoCl2·6H2O 0.025mg/L, CuSO4·5H2O
0.025mg/L, inositol 10mg/L, niacin 0.05mg/L, thiamine hydrochloride 0.01mg/L, puridoxine hydrochloride 0.05mg/L)+6-
BA0.3mg/L+NAA 0.01mg/L+ glucose 20g/L+ carragheen 5.5g/L；
Wherein, in step (2.2), it be light application time is 12h/d that Bulgarian essential oil rose squamous subculture, which obtains condition of culture,
Intensity of illumination 2000-3000lux, cultivation temperature are 26 ± 2 DEG C；
As described in Table 2, pH is adjusted to 5.9 for growth performance in each subculture medium:
Table 2
In table 2, the dosage of a great number of elements, calcium salt, molysite is improved, 1.5 times of maximum concentration to former dosage, is successfully improved
The color of plant, shows as green.
6-BA and NAA combined treatment, when 6-BA is improved using concentration, the sprouting amount of clump bud is also increased accordingly, but group
It is thinner and more delicate to train seedling, when being applied to production, reduces the ratio of effective seedling, loses more than gain；When 6-BA be 0.3mg/L,
When NAA0.01mg/L, multiplied ratio can reach 4.5, and robust plant, and the effective seedling ratio for carrying out squamous subculture reaches
99% or more, improve the utilization rate of seedling, and be conducive to next stage to take root operation.
(2.3) after cultivating 35d, plant longitudinal growth plant height is sprouted in 3.5cm or more, plant lateral bud, and lower part has one
Lateral budding is long to form single size about 2cm, and statistics multiplied ratio averagely reaches 1:4.5.
A specific embodiment of the invention is illustrated above, but the present invention is not limited thereto, without departing from
Spirit of the invention, the present invention can also have various change.
Claims (2)
1. a kind of subculture method of Bulgaria essential oil rose, it is characterised in that: the following steps are included:
(1) experimental material must select: select that growing way is neat, plant height 2.5-4.0cm, subculture cycle are 30-35 days, leaf color
In the Bulgarian Rosa rugosa tissue-cultured seedling of green or peak green as squamous subculture material；
(2) specific experiment step:
(2.1) cutting of material: excision blade retains terminal bud lobus cardiacus, and petiole stays 0.1-0.2cm, is cut into the stem of 0.5-0.8cm
Section；
(2.2) Bulgarian essential oil rose squamous subculture；Terminal bud and stem section separate inoculation, are equably inoculated into subculture medium 1
In number；
(2.3) after cultivating 35d, plant longitudinal growth plant height is sprouted in 3.5cm or more, plant lateral bud, and a lateral bud is arranged at lower part
Growth forms single size about 2cm, and statistics multiplied ratio averagely reaches 1:4.5；
No. 1 KNO of subculture medium in step (2.2)31.9-2.5g/L, NH4NO31.65-2.1g/L MgSO4·7H2O
0.37-0.48g/L, KH2PO40.17-0.22g/L, CaCl2·2H2O 0.44-0.57g/L, Na2EDTA·2H2O 0.041-
0.056g/L,FeSO4·7H2O 0.03-0.041g/L,MnSO4·H2O 16.9mg/L, H3BO36.2mg/L, ZnSO4·7H2O
8.6mg/L, KI 0.83mg/L, Na2MoO4·2H2O 0.25mg/L, CoCl2·6H2O 0.025mg/L, CuSO4·5H2O
0.025mg/L, inositol 10mg/L, niacin 0.05mg/L, thiamine hydrochloride 0.01mg/L, puridoxine hydrochloride 0.05mg/L are added
6-BA 0.3mg/L, NAA 0.01mg/L, glucose 20g/L+ carragheen 5.5g/L.
2. a kind of subculture method of Bulgarian essential oil rose according to claim 1, it is characterised in that: step
(2.2) in, the condition of culture of Bulgarian essential oil rose squamous subculture is that light application time is 12h/d, intensity of illumination 2000-
3000lux, cultivation temperature are 26 ± 2 DEG C.
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| CN102027881A (en) * | 2010-11-10 | 2011-04-27 | 天津滨海国际花卉科技园区股份有限公司 | Method for overcoming yellowing of leaves of tissue culture seedlings of Rosa damascena |
| CN103960133A (en) * | 2014-05-27 | 2014-08-06 | 昆明学院 | Method for tissue culture and rapid propagation of Rosa rugosa Thunb. |
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| CN102027881A (en) * | 2010-11-10 | 2011-04-27 | 天津滨海国际花卉科技园区股份有限公司 | Method for overcoming yellowing of leaves of tissue culture seedlings of Rosa damascena |
| CN103960133A (en) * | 2014-05-27 | 2014-08-06 | 昆明学院 | Method for tissue culture and rapid propagation of Rosa rugosa Thunb. |
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| 保加利亚精油玫瑰的离体起始培养与快速繁殖;康冰等;《西北林学院学报》;20041231;第19卷(第1期);第70-72页，尤其是第1.3、2.5节 * |
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