CN106718896A - A kind of subculture method of Bulgarian essential oil rose - Google Patents
A kind of subculture method of Bulgarian essential oil rose Download PDFInfo
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- CN106718896A CN106718896A CN201611119737.0A CN201611119737A CN106718896A CN 106718896 A CN106718896 A CN 106718896A CN 201611119737 A CN201611119737 A CN 201611119737A CN 106718896 A CN106718896 A CN 106718896A
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- 241000220317 Rosa Species 0.000 title claims abstract description 73
- 239000000341 volatile oil Substances 0.000 title claims abstract description 24
- 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 13
- 241001582888 Lobus Species 0.000 claims abstract description 5
- 238000011081 inoculation Methods 0.000 claims abstract description 5
- 239000012879 subculture medium Substances 0.000 claims abstract description 5
- 235000000659 Rosa rugosa Nutrition 0.000 claims description 9
- 240000006066 Rosa rugosa Species 0.000 claims description 9
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 claims description 8
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 claims description 8
- 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
- 238000009395 breeding Methods 0.000 claims description 6
- 230000001488 breeding effect Effects 0.000 claims description 6
- 239000008103 glucose Substances 0.000 claims description 6
- 238000012136 culture method Methods 0.000 claims description 5
- 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
- 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
- 230000015572 biosynthetic process Effects 0.000 abstract description 3
- 230000034303 cell budding Effects 0.000 abstract description 3
- 206010053759 Growth retardation Diseases 0.000 abstract description 2
- 206010023126 Jaundice Diseases 0.000 abstract description 2
- 231100000001 growth retardation Toxicity 0.000 abstract description 2
- 238000011534 incubation Methods 0.000 abstract description 2
- 239000004615 ingredient Substances 0.000 abstract description 2
- 235000016709 nutrition Nutrition 0.000 abstract description 2
- 238000011160 research Methods 0.000 description 7
- 239000005556 hormone Substances 0.000 description 6
- 229940088597 hormone Drugs 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 229920001817 Agar Polymers 0.000 description 5
- 239000008272 agar Substances 0.000 description 5
- 238000005516 engineering process 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
- 239000005720 sucrose Substances 0.000 description 4
- 238000012090 tissue culture technique Methods 0.000 description 4
- 238000012549 training Methods 0.000 description 4
- 230000006872 improvement Effects 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 238000011282 treatment 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
- 238000009825 accumulation Methods 0.000 description 2
- 230000009286 beneficial effect Effects 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
- 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
- 235000004789 Rosa xanthina 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
- 230000001413 cellular effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 239000000039 congener Substances 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
- 239000002609 medium Substances 0.000 description 1
- 239000013028 medium composition Substances 0.000 description 1
- 230000015689 metaplastic ossification Effects 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000000750 progressive effect 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 Bulgarian essential oil rose, comprises the following steps：(1) experiment material must be selected；(2) specific experiment step：(2.1) acquisition of aseptic explant：Excision blade, retains terminal bud lobus cardiacus, and petiole stays 0.1 0.2cm, cuts 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 culture 35d, plant longitudinal growth plant height is sprouted in more than 3.5cm, plant lateral bud, and a lateral budding single size about 2cm of formation long is arranged at bottom, and statistics multiplied ratio averagely reaches 1:4.5.Nutritional ingredient of the present invention needed for rose squamous subculture incubation is adjusted, and solves the phenomenon of plant jaundice growth retardation in rose tissue-cultured seedling Subculture；And improve the multiplied ratio of rose tissue-cultured seedling.The tissue culture system of rose is successfully established, rose factorial praluction is realized.
Description
Technical field
The present invention relates to a kind of tissue culture method, and in particular to a kind of subculture method of Bulgarian essential oil rose, category
In biological technical field.
Background technology
Bulgarian Rosa rugosa belongs to the rose family, Rosa, machaka.It is fuel-displaced because Bulgarian Rosa rugosa belongs to international odor type
Rate is up to 3/10000ths. six, is the optimal kind for extracting Rosa Damascana and production rose (fresh flower water).Bulgarian rose
Rare yield greatly, can reach 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 optimal kind of rose flavored juice, thus quilt
Plantation extensively is used to extract attar of rose, with economic worth very high.The rose industry of Bulgaria is existing more than 300 years to be gone through
History, have the good reputation of " rose kingdom ".The rose produced with Bulgarian Rosa rugosa fresh flower, per kilogram price is beautiful up to 1000
Unit.The Rosa Damascana extracted from Bulgarian Rosa rugosa fresh flower, per kilogram price is up to 7000~8000 dollars, have " liquid
The title of gold ".
Bulgarian Rosa rugosa strong adaptability, cultivation management is easy, high financial profit.To the north of Yangtze River in China and on the south Beijing
Qu Junke is cultivated.It is a new industry with broad mass market prospect and huge exploitation potential.
Existing investigative technique：
In the existing report on rose tissue cultures, rose tissue cultures are carried out by point of penetration of hormone combinations more
Research.Deng Yanhong etc. exists《Rose Study on tissue culture》It is white using MS+6-BA2.0mg/L+NAA 0.01mg/L+3% in report
Sugar+0.4g/L agar this formula carries out the Multiplying culture of rose, and proliferation times can reach more than 6；
Zhao Yipeng etc. exists《The tissue culture technique of rose》The similar formula of class is also adopted by research, only to the concentration of hormone
Adjusted, minimal medium selects MS, and the excursion 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. exists《Rose tissue culture rapid propagating technology》Middle proposition, rose is in subculture medium MS+6-BA3.0mg/L
Growth coefficient highest in+NAA 0.1mg/L, can reach 4；
In keep it is superfine studied as test material with purple branch rose when《The research of purple branch rose tissue cultures》, using MS
+ 1.8mg/l 6-BA+0.08mg/L NAA+0.4mg/l GA3 effects are fine；
《Spend Rose plants tissue cultures pre-test in vain》With《The research of safflower Rose plants tissue cultures》Respectively spending rose in vain
Rare and safflower rose is formulated MS+6-BA 3.0mg/L+NAA 0.2mg/ as test material, squamous subculture stage using identical
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》With《The Study on tissue culture of black red rose》Worked as from Yunnan
The edible rose kind " Mo Hong " on ground is respectively MS+6-BA3.0mg/L+NAA as test material, the culture formula of multiplicative stage
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, increase
Grow coefficient and be respectively 3.5 and 3.517.
《Essential oil rose tissue culture technique is studied》It is material with " Damascus " rose in one text, is basic training with MS
Base is supported, by Orthogonal Experiment and Design, the conclusion for obtaining is：Hormone is cultivated with 6BA1.0mg/L and NAA0.01mg/l treatment
Carbon source selection white granulated sugar consumption is that 40g/L is best to the cultivation effect of rose in base.
《Damask Rose tissue culture technique is studied》The suitable Damask Rose Multiplying culture drawn 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.
《Rose tissue cultures and fast breeding technique experimental study》Entered with breeding " Bulgaria " rose that Gansu province is introduced
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.
Studies have reported that weak point：
The studies above report rose material used can be divided into be viewed and admired with rose, edible rose and smart rose for oil.Plant
The theoretical foundation of tissue culture technique is cellular omnipotency theoretical, even and congener, because genotype is different, tissue cultures
In squamous subculture technology used it is also variant.Aromatic substance is abundant compared with other 2 class roses in smart rose for oil body, therefore directly
Connect the demand having influence on during its tissue culture for each nutriment and Auto-regulator.It is many in culture medium in above-mentioned report
Cultivated with 6-BA (concentration is more than 2.0) collocation NAA of high concentration, 6-BA many generations may reduce tissue-cultured seedling using easily accumulation
Differentiation capability, while carrying out group training research by material of " Bulgaria " rose, is carried out using 3 kinds of formula interactions, and this is for work
Factory's metaplasia is produced and easily causes managerial confusion, is difficult to carry out the control of production technology.
The present invention is test material with " Bulgaria " rose, and more than 4.5 are can reach using one kind formula growth coefficient, into
Work(realizes the factorial praluction of rose.
The content of the invention
It is an object of the present invention to provide a kind of subculture method of Bulgarian essential oil rose, to overcome existing skill
Disadvantages mentioned above and deficiency existing for art.
This experiment is directed to a kind of Subculture of Bulgarian essential oil rose, solves a kind of Bulgarian essential oil rose
Multiplied ratio is low in rare tissue-culturing rapid propagation, and tissue-cultured seedling turns to be yellow and vitrified problem, so as to realize Bulgarian essential oil rose tissue culture
Factorial praluction.
The technical problem solved required for of the invention, can be achieved through the following technical solutions：
A kind of quick breeding by group culture method of Bulgarian essential oil rose, it is characterised in that：Comprise the following steps：
(1) experiment material must be selected：Select that growing way is neat, plant height be 2.5-4.0cm, subculture cycle be 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, cuts 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, improvement MS (KNO 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, nicotinic acid 0.05mg/L, thiamine hydrochloride 0.01mg/L, puridoxine hydrochloride 0.05mg/L)+6-
BA0.3mg/L+NAA 0.01mg/L+ glucose 20g/L+ carragheens 5.5g/L；
Wherein, in step (2.2), Bulgarian essential oil rose squamous subculture obtains condition of culture for light application time is 12h/d,
Intensity of illumination 2000-3000lux, cultivation temperature is 26 ± 2 DEG C；
(2.3) after culture 35d, plant longitudinal growth plant height is sprouted in more than 3.5cm, plant lateral bud, and bottom has one
Lateral budding single size about 2cm of formation long, statistics multiplied ratio averagely reaches 1:4.5.
Beneficial effects of the present invention：
The present invention is adjusted for the nutritional ingredient needed for rose squamous subculture incubation, improve a great number of elements,
The consumption of molysite, calcium salt, carbon source uses glucose instead, solves the phenomenon of plant jaundice growth retardation during rose tissue-cultured seedling；
The combined treatment of the 6-BA and NAA of low concentration is used simultaneously, is reduced because the too high and many generation accumulation of hormone causes 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 composition and uses suitable hormone combinations, is matched somebody with somebody using one group of Multiplying culture
Side, is successfully established the tissue culture system of rose, realizes rose factorial praluction.
Specific embodiment
Below in conjunction with specific embodiment, progressive explanation is made to the present invention.It should be understood that following examples are merely to illustrate this hair
It is bright not for limit the scope of the present invention.
Embodiment 1
A kind of quick breeding by group culture method of Bulgarian essential oil rose, comprises the following steps：
(1) experiment material must be selected：Select that growing way is neat, plant height be 2.5-4.0cm, subculture cycle be 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, cuts 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, improvement MS (KNO 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, nicotinic acid 0.05mg/L, thiamine hydrochloride 0.01mg/L, puridoxine hydrochloride 0.05mg/L)+6-
BA0.3mg/L+NAA 0.01mg/L+ glucose 20g/L+ carragheens 5.5g/L；
Wherein, in step (2.2), Bulgarian essential oil rose squamous subculture obtains condition of culture for light application time is 12h/d,
Intensity of illumination 2000-3000lux, cultivation temperature is 26 ± 2 DEG C；
Growth performance in each subculture medium is as described in Table 1, and pH is adjusted to 5.9：
Table 1
Table 1 test result indicate that：Compared with white granulated sugar, with the addition of in the culture medium of glucose, rose tissue-cultured seedling grows
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 Bulgarian essential oil rose, comprises the following steps：
(1) experiment material must be selected：Select that growing way is neat, plant height be 2.5-4.0cm, subculture cycle be 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, cuts 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, improvement MS (KNO 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, nicotinic acid 0.05mg/L, thiamine hydrochloride 0.01mg/L, puridoxine hydrochloride 0.05mg/L)+6-
BA0.3mg/L+NAA 0.01mg/L+ glucose 20g/L+ carragheens 5.5g/L；
Wherein, in step (2.2), Bulgarian essential oil rose squamous subculture obtains condition of culture for light application time is 12h/d,
Intensity of illumination 2000-3000lux, cultivation temperature is 26 ± 2 DEG C；
Growth performance in each subculture medium is as described in Table 2, and pH is adjusted to 5.9：
Table 2
In table 2, a great number of elements, calcium salt, the consumption of molysite are improved, 1.5 times of maximum concentration to former consumption, successfully improved
The color of plant, shows as green.
6-BA and NAA combined treatments, when 6-BA concentrations are improved, the sprouting amount of clump bud is also improved therewith, but group
Training seedling is thinner and more delicate, when being applied to production, reduces the ratio of effective seedling, loses more than gain；When 6-BA is 0.3mg/L,
During NAA0.01mg/L, multiplied ratio can reach 4.5, and robust plant, and the effective seedling ratio for carrying out squamous subculture reaches
More than 99%, improve the utilization rate of seedling, and take root operation beneficial to next stage.
(2.3) after culture 35d, plant longitudinal growth plant height is sprouted in more than 3.5cm, plant lateral bud, and bottom has one
Lateral budding single size about 2cm of formation long, statistics multiplied ratio averagely reaches 1:4.5.
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 (3)
1. a kind of subculture method of Bulgarian essential oil rose, it is characterised in that：Comprise the following steps：
(1) experiment material must be selected：Select that growing way is neat, plant height is that 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, cuts 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 (MG-1)；
(2.3) after culture 35d, plant longitudinal growth plant height is sprouted in more than 3.5cm, plant lateral bud, and a lateral bud is arranged at bottom
Growth forms single size about 2cm, and statistics multiplied ratio averagely reaches 1:4.5.
2. the quick breeding by group culture method of a kind of Bulgarian essential oil rose according to claim 1, it is characterised in that：Step
Suddenly MS (KNO are improved in (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, nicotinic acid 0.05mg/L, thiamine hydrochloride 0.01mg/L, puridoxine hydrochloride 0.05mg/L, additional 6-BA0.3mg/
L, NAA0.01mg/L, glucose 20g/L+ carragheens 5.5g/L.
3. the subculture method of a kind of Bulgarian essential oil rose according to claim 1, it is characterised in that：Step
(2.2) in, Bulgarian essential oil rose squamous subculture obtains condition of culture for light application time is 12h/d, intensity of illumination 2000-
3000lux, cultivation temperature is 26 ± 2 DEG C.
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| Publication number | Priority date | Publication date | Assignee | Title |
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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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| Publication number | Priority date | Publication date | Assignee | Title |
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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. |
Non-Patent Citations (1)
| Title |
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| 康冰等: "保加利亚精油玫瑰的离体起始培养与快速繁殖", 《西北林学院学报》 * |
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