CN107568062A - A kind of composite reagent and its application in the bletilla striata is cultivated - Google Patents

A kind of composite reagent and its application in the bletilla striata is cultivated Download PDF

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Publication number
CN107568062A
CN107568062A CN201710836183.4A CN201710836183A CN107568062A CN 107568062 A CN107568062 A CN 107568062A CN 201710836183 A CN201710836183 A CN 201710836183A CN 107568062 A CN107568062 A CN 107568062A
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mother liquor
liter
iii
composite reagent
bletilla striata
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胡俊
高丽
高睿琦
周丽
石俊
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Flowers And Trees Group Co Ltd Of Beijing Figured Township
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Flowers And Trees Group Co Ltd Of Beijing Figured Township
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Abstract

The invention discloses a kind of composite reagent, its drip irrigation device is to include mother liquor I, mother liquor II, mother liquor III and mother liquor IV, wherein, every liter of mother liquor I：KNO3 30‑46g、NH4NO3 27‑39g、CaCl2·2H2O 8‑9.6g、MgSO4·7H2O 7‑7.8g、KH2PO43‑3.8g；Every liter of mother liquor II：KI、H3BO3 、MnSO4·4H2O、ZnSO4·7H2O、Na2MoO4·2H2O、CuSO4·5H2O、CoCl2·6H2O；Every liter of mother liquor III：Inositol, nicotinic acid, pyridoxine hydrochloride, thiamine hydrochloride, glycine；Every liter of mother liquor IV：FeSO4·7H2O、Na2·EDTA·2H2O, the a variety of psychological needs for meeting flowers under reproductive status are reached, the collocation selection of various trace elements promotes flowers fast-growth in incubation, and breeding coefficient is high, can completely inherits maternal good characteristic, reaches the expectation of people's flowers industrialized cultivation.

Description

A kind of composite reagent and its application in the bletilla striata is cultivated
Technical field
The present invention relates to bletilla striata field of planting, more particularly to a kind of composite reagent and its application in the bletilla striata is cultivated.
Background technology
In vainSplendid achnatherum, also known asIt is even and careless, Gan Gen,Give free of charge、Indocalamus is blue、Zhu LanDeng perennial herbBulbous plant（Root tuber）, plant height 18-60 centimetres.It is mainly distributed on China, Japan and Upper Myanmar.MainlyFlorescenceIn spring, but according to the difference of various regions weather, Late Winter Solstice early summer may all bloom.The bletilla striata has extensive medical value and Ornamental value.It is mainly used in astringing to arrest bleeding, detumescence life Flesh.Flower has the colors such as purplish red, white, blue, yellow and powder, can potted plant indoor appreciation, can also intersperse in more cover flower stand, flower border or front yard One jiao of institute.
The existing Chinese patent for referring to Publication No. CN104255494A, it discloses a kind of iris tissue culture culture Base, it is related to iris planting technology field, it is characterised in that：Including following media, inducing culture, precious No.1 2.8g/L is spent + inositol 0.1g/L+ coconut juices 100mL/L+BA4.0mg/L+LAde5.0mg/L+NAA1.0mg/L；Proliferated culture medium, 1/3MS+ fleshes Alcohol 0.1g/L+ citric acid 0.03g/L+ carbon dust 1g/L+ coconut juices 100mL/L；Root media, 1/2MS+ bananas homogenate 100mL/L+ Carbon dust 1g/L+ABT1 root-inducing powders 1.0mg/L；It is reasonable that the present invention prepares, and inductivity is high, and plant growing way can be made strong, and growth is fast, leaf Piece is sturdy.
The extensive medicinal and Ornamental value of the bletilla striata allows many plantation enterprise more urgent need one kind can be by bletilla seed Plant efficient, industrialized means.
The content of the invention
It is an object of the invention to provide a kind of composite reagent, serves a variety of physiology need for meeting flowers under reproductive status Ask, the collocation of various trace elements selection promotes flowers fast-growth in incubation, and breeding coefficient is high, can completely after The good characteristic of female parent is held, reaches the expectation of people's flowers industrialized cultivation.
The present invention above-mentioned technical purpose technical scheme is that：A kind of composite reagent, includes Mother liquor I, mother liquor II, mother liquor III and mother liquor IV,
Wherein, every liter of mother liquor I：KNO3 30-46g、NH4NO3 27-39g、CaCl2·2H2O 8-9.6g、MgSO4·7H2O 7- 7.8g、KH2PO43-3.8g；
Every liter of mother liquor II：KI 0.16-0.172g、H3BO3 1.2-1.28g、MnSO4·4H2O 4.4-4.52g、ZnSO4·7H2O 1.7-1.74g、Na2MoO4·2H2O 0.03-0.07g、CuSO4·5H2O 0.03-0.07g、CoCl2·6H2O 0.03- 0.07g；
Every liter of mother liquor III：Inositol 16-24g, nicotinic acid 0.07-0.13g, pyridoxine hydrochloride 0.07-0.13g, thiamine hydrochloride 0.01-0.03g/L, glycine 0.2-0.6g/L；
Every liter of mother liquor IV：FeSO4·7H2O 5.5-5.62g/L、Na2·EDTA·2H2O 7.46g/L。
Pass through above-mentioned technical proposal, the main plantation for meeting flowers of the modulation of composite reagent use, and pass through above-mentioned four The adjustment of proportion can meet the needs of flowers are under a variety of physiological status between the mixing of kind solution and wherein each component, The collocation selection of various trace elements promotes flowers fast-growth in incubation, and breeding coefficient is high, and can completely is inherited Maternal good characteristic, reaches the expectation of people's flowers industrialized cultivation.
Preferably, also include the growth promoter for promoting plant growth and division.
Preferably, growth promoter includes 6-BA and the Plant Hormones of IAA two.
Preferably, 6-BA and IAA weight ratio is 1:1, wherein 6-BA and IAA concentration are 500mg/L.
Pass through above-mentioned technical proposal, two kinds of being used cooperatively for bio-hormone can stably allow flowers to carry out fast-growth, Promote the division of cell simultaneously, have preferable effect for some breedings and the slow plant of growth rate.
Preferably, also include nutritious supplementary pharmaceutical, every liter of nutritious supplementary pharmaceutical include pyrolkigneous liquid 60mL, tomato juice 240mL, Ginkgo liquid extraction liquid 75ml, sucrose 105g, agar powder 15g.
Pass through above-mentioned technical proposal, the selection of nutritious supplementary pharmaceutical can be very good to promote the division and growth of plant, simultaneously The pyrolkigneous liquid of addition has certain bacteriostasis property, reduces the pollution that culture medium is subject to when in use.
The purpose of the present invention two：A kind of application of composite reagent in bletilla striata Subculture.
The purpose of the present invention three：A kind of bletilla striata subculture medium, include the 45-55ml of mother liquor I in every liter of culture medium, it is female Liquid II, mother liquor III and the 3-7ml of mother liquor IV, growth promoter 0.06-0.1ml, nutritious supplementary pharmaceutical 200-400ml, spend precious 2.5- 4.5g, agar powder 5.5-7.5g.
Pass through above-mentioned technical proposal, the composite reagent is applied good effect, the bletilla striata in bletilla striata Subculture Seed speed of germinating is fast, and germination rate is high；Seedling is short strong after squamous subculture, has protocorm during bottle outlet, survival rate height after emergence.
The purpose of the present invention four：A kind of preparation method of bletilla striata subculture medium, comprises the following steps：
S1：Mother liquor I, mother liquor II, mother liquor III, mother liquor IV and nutritious supplementary pharmaceutical and each some liters of growth promoter are prepared, and are deposited It is stored in refrigerating chamber；
S2：When needing to use culture medium, according to proportioning by mother liquor I, mother liquor II, mother liquor III, mother liquor IV and nutritious supplementary pharmaceutical It is sufficiently mixed to obtain mixed liquor A with growth promoter, NaOH or the HCl regulation that then concentration is 0.5mol/L are mixed Liquid A pH value is closed, is ended when ph values reach 5.8；
S3：It is dispensed into every bottle of 80ML/ blake bottle, is put into high-pressure sterilizing pot, sterilizes 25min, high-pressure sterilizing pot work temperature Degree 120 DEG C, pressure 120kPa, after the completion of refrigeration place it is stand-by.
Pass through above-mentioned technical proposal, by above-mentioned preparation method prepare can with the bacterial content in stability contorting culture medium with And acid-base value, the specific living environment requirement of bletilla striata growth is grasped well, and the bletilla striata can be allowed to possess and be more suitable for itself life Long external environment.
In summary, the invention has the advantages that：
1st, the addition of pyrolkigneous liquid can supplement substantial amounts of trace element, while the addition of pyrolkigneous liquid also has sterilization, antibacterial effect, When the bletilla striata is planted in the industrialization of large area, the environment for cultivating the bletilla striata will necessarily slightly below in the lab, training now Foster base is easy to be contaminated, and the addition of pyrolkigneous liquid can sterilize, be antibacterial, reduces to cultivate caused by culture medium is contaminated and loses Lose, reduce the risk of culture, the plantation of the bletilla striata can be more prone to；
2nd, the culture medium that the design of tissue culture mode can allow the bletilla striata to use can be allocated largely in advance, just carry out when it is desired to be used Mixing, the problem of so as to save cost of some workers itself, while it is also convenient for factory and is prepared.
Embodiment
The present invention is described in further detail below.
All substances in the present invention are both from market.
Embodiment 1：Composite reagent is configured first, includes mother liquor I, mother liquor II, mother liquor III and mother liquor IV,
Wherein, every liter of mother liquor I：KNO3 46g、NH4NO339g、CaCl2·2H2O 9.6g、MgSO4·7H2O 7.8g、KH2PO4 3.8g；
Every liter of mother liquor II：KI 0.172g、H3BO31.28g、MnSO4·4H2O 4.52g、ZnSO4·7H2O 1.74g、 Na2MoO4·2H2O 0.07g、CuSO4·5H2O 0.07g、CoCl2·6H2O 0.07g；
Every liter of mother liquor III：Inositol 24g, nicotinic acid 0.13g, pyridoxine hydrochloride 0.13g, thiamine hydrochloride 0.03g, glycine 0.6g；
Every liter of mother liquor IV：FeSO4·7H2O 5.62g/L、Na2·EDTA·2H2O 7.46g/L。
Growth promoter, include the grade weight 6-BA and IAA that concentration is 500mg/L；
Nutritious supplementary pharmaceutical, every liter of nutritious supplementary pharmaceutical include pyrolkigneous liquid 60mL, tomato juice 240mL, ginkgo liquid extraction liquid 75ml, sucrose 105g, agar powder 15g.
Bletilla striata subculture medium and making：
S1：The mother liquor I as in above-mentioned composite reagent prefabricated first, mother liquor II, mother liquor III and mother liquor IV, growth promoter, nutrition Replenishers, and prepare to spend precious and agar powder to be refrigerated；
S2：The 55ml of mother liquor I, mother liquor II, mother liquor III, the 7ml of mother liquor IV, nutritious supplementary pharmaceutical are chosen from prefabricated composite reagent 400ml and growth promoter 0.1ml, precious 4.5g, agar powder 7.5g are spent, various solution are sufficiently mixed to obtain mixed liquor A, so Concentration is that 0.5mol/L NaOH or HCl adjust the pH value of mixed liquor A afterwards, is ended when ph values reach 5.8；
S3：It is dispensed into every bottle of 80ML/ blake bottle, is put into high-pressure sterilizing pot, sterilizes 25min, high-pressure sterilizing pot work temperature Degree 120 DEG C, pressure 120kPa, after the completion of refrigeration place it is stand-by.
Embodiment 2：Composite reagent is configured first, includes mother liquor I, mother liquor II, mother liquor III and mother liquor IV,
Wherein, every liter of mother liquor I：KNO3 30g、NH4NO3 27g、CaCl2·2H2O 8g、MgSO4·7H2O 7g、KH2PO43g；
Every liter of mother liquor II：KI 0.16g、H3BO3 1.2g、MnSO4·4H2O 4.4g、ZnSO4·7H2O 1.7g、Na2MoO4· 2H2O 0.03g、CuSO4·5H2O 0.03g、CoCl2·6H2O 0.03g；
Every liter of mother liquor III：Inositol 16g, nicotinic acid 0.07g, pyridoxine hydrochloride 0.07g, thiamine hydrochloride 0.01g, glycine 0.2g；
Every liter of mother liquor IV：FeSO4·7H2O 5.5g、Na2·EDTA·2H2O 7.46g。
Growth promoter, include 6-BA and IAA that concentration is 500mg/L；
Nutritious supplementary pharmaceutical, every liter of nutritious supplementary pharmaceutical include pyrolkigneous liquid 60mL, tomato juice 240mL, ginkgo liquid extraction liquid 75ml, sucrose 105g, agar powder 15g.
Bletilla striata subculture medium and making：
S1：The mother liquor I as in above-mentioned composite reagent prefabricated first, mother liquor II, mother liquor III and mother liquor IV, growth promoter, battalion Replenishers are supported, and prepare to spend precious and agar powder to be refrigerated；
S2：The 45ml of mother liquor I, mother liquor II, mother liquor III, the 3ml of mother liquor IV, nutritious supplementary pharmaceutical are chosen from prefabricated composite reagent 200ml and growth promoter 0.06ml, precious 2.5g is spent, the various solution of agar powder 5.5g are sufficiently mixed to obtain mixed liquor A, so Concentration is that 0.5mol/L NaOH or HCl adjust the pH value of mixed liquor A afterwards, is ended when ph values reach 5.8；
S3：It is dispensed into every bottle of 80ML/ blake bottle, is put into high-pressure sterilizing pot, sterilizes 25min, high-pressure sterilizing pot work temperature Degree 120 DEG C, pressure 120kPa, after the completion of refrigeration place it is stand-by.
Embodiment 3：Composite reagent is configured first, includes mother liquor I, mother liquor II, mother liquor III and mother liquor IV,
Wherein, mother liquor I：KNO3 38g、NH4NO3 33g、CaCl2·2H2O 8.8g、MgSO4·7H2O 7.4g、KH2PO4 3.4g；
Mother liquor II：KI 0.166g、H3BO3 1.24g、MnSO4·4H2O 4.46g、ZnSO4·7H2O 1.72g、Na2MoO4· 2H2O 0.05g、CuSO4·5H2O 0.005g、CoCl2·6H2O 0.005g；
Mother liquor III：Inositol 20g, nicotinic acid 0.1g, pyridoxine hydrochloride 0.1g, thiamine hydrochloride 0.02g, glycine 0.4g；
Mother liquor IV：FeSO4·7H2O 5.56g、Na2·EDTA·2H2O 7.46g；
Growth promoter, include 6-BA and IAA that concentration is 500mg/L；
Nutritious supplementary pharmaceutical, every liter of nutritious supplementary pharmaceutical include pyrolkigneous liquid 60mL, tomato juice 240mL, ginkgo liquid extraction liquid 75ml, sucrose 105g, agar powder 15g.
Bletilla striata subculture medium and making：
S1：The mother liquor I as in above-mentioned composite reagent prefabricated first, mother liquor II, mother liquor III and mother liquor IV, growth promoter, battalion Replenishers are supported, and prepare to spend precious and agar powder to be refrigerated；
S2：The 50ml of mother liquor I, mother liquor II, mother liquor III, the 5ml of mother liquor IV, nutritious supplementary pharmaceutical are chosen from prefabricated composite reagent 300ml and growth promoter 0.08ml, precious 3.5g, agar powder 6.5g are spent, various solution are sufficiently mixed to obtain mixed liquor A, Then concentration is that 0.5mol/L NaOH or HCl adjust the pH value of mixed liquor A, is ended when ph values reach 5.8；
S3：It is dispensed into every bottle of 80ML/ blake bottle, is put into high-pressure sterilizing pot, sterilizes 25min, high-pressure sterilizing pot work temperature Degree 120 DEG C, pressure 120kPa, after the completion of refrigeration place it is stand-by.
Embodiment 4：Composite reagent is configured first, includes mother liquor I, mother liquor II, mother liquor III and mother liquor IV,
Wherein, every liter of mother liquor I：KNO3 30g、NH4NO3 27g、CaCl2·2H2O 8g、MgSO4·7H2O 7g、KH2PO43g；
Every liter of mother liquor II：KI 0.16g、H3BO3 1.2g、MnSO4·4H2O 4.4g、ZnSO4·7H2O 1.7g、Na2MoO4· 2H2O 0.03g、CuSO4·5H2O 0.03g、CoCl2·6H2O 0.03g；
Every liter of mother liquor III：Inositol 16g, nicotinic acid 0.07g, pyridoxine hydrochloride 0.07g, thiamine hydrochloride 0.01g, glycine 0.2g；
Every liter of mother liquor IV：FeSO4·7H2O 5.5g、Na2·EDTA·2H2O 7.46g/L。
Growth promoter, include 6-BA and IAA that concentration is 500mg/L；
Nutritious supplementary pharmaceutical, every liter of nutritious supplementary pharmaceutical include pyrolkigneous liquid 60mL, tomato juice 240mL, ginkgo liquid extraction liquid 75ml, sucrose 105g, agar powder 15g.
Bletilla striata subculture medium and making：
S1：The mother liquor I as in above-mentioned composite reagent prefabricated first, mother liquor II, mother liquor III and mother liquor IV, growth promoter, battalion Replenishers are supported, and prepare to spend precious and agar powder to be refrigerated；
S2：The 55ml of mother liquor I, mother liquor II, mother liquor III, the 7ml of mother liquor IV, nutritious supplementary pharmaceutical are chosen from prefabricated composite reagent 400ml and growth promoter 0.1ml, precious 3.5g, agar powder 6.5g are spent, various solution are sufficiently mixed to obtain mixed liquor A, so Concentration is that 0.5mol/L NaOH or HCl adjust the pH value of mixed liquor A afterwards, is ended when ph values reach 5.8；
S3：It is dispensed into every bottle of 80ML/ blake bottle, is put into high-pressure sterilizing pot, sterilizes 25min, high-pressure sterilizing pot work temperature Degree 120 DEG C, pressure 120kPa, after the completion of refrigeration place it is stand-by.
Embodiment 5：Composite reagent is configured first, includes mother liquor I, mother liquor II, mother liquor III and mother liquor IV：
Wherein, every liter of mother liquor I：KNO346g、NH4NO3 39g、CaCl2·2H2O 9.6g、MgSO4·7H2O 7.8g、KH2PO4 3.8g；
Every liter of mother liquor II：KI 0.172g、H3BO3 1.28g、MnSO4·4H2O 4.52g、ZnSO4·7H2O 1.74g、 Na2MoO4·2H2O 0.07g、CuSO4·5H2O0.07g、CoCl2·6H2O 0.07g；
Every liter of mother liquor III：Inositol 24g, nicotinic acid 0.13g, pyridoxine hydrochloride 0.13g, thiamine hydrochloride 0.03g, glycine 0.6g；
Every liter of mother liquor IV：FeSO4·7H2O 5.62g、Na2·EDTA·2H2O 7.46g。
Growth promoter, include 6-BA and IAA that concentration is 500mg/L；
Nutritious supplementary pharmaceutical, every liter of nutritious supplementary pharmaceutical include pyrolkigneous liquid 60mL, tomato juice 240mL, ginkgo liquid extraction liquid 75ml, sucrose 105g, agar powder 15g.
Bletilla striata subculture medium and making：
S1：The mother liquor I as in above-mentioned composite reagent prefabricated first, mother liquor II, mother liquor III and mother liquor IV, growth promoter, battalion Replenishers are supported, and prepare to spend precious and agar powder to be refrigerated；
S2：The 45ml of mother liquor I, mother liquor II, mother liquor III, the 3ml of mother liquor IV, nutritious supplementary pharmaceutical are chosen from prefabricated composite reagent 200ml and growth promoter 0.06ml, precious 3.5g, agar powder 6.5g are spent, various solution are sufficiently mixed to obtain mixed liquor A, Then concentration is that 0.5mol/L NaOH or HCl adjust the pH value of mixed liquor A, is ended when ph values reach 5.8；
S3：It is dispensed into every bottle of 80ML/ blake bottle, is put into high-pressure sterilizing pot, sterilizes 25min, high-pressure sterilizing pot work temperature Degree 120 DEG C, pressure 120kPa, after the completion of refrigeration place it is stand-by.
Bletilla striata squamous subculture overall process
Step 1：The processing of explant, the Fruit pod of the bletilla striata is selected all to call explant below as explant, rinsed with flowing water Several times, dry standby；
Step 2：Explant is first with 75% alcohol disinfecting 1min, and with sterile water wash 2 times, explant is again with 2% sodium hypochlorite Solution disinfection 10min, with sterile water wash 3 times, the moisture on explant surface is blotted with the filter paper sterilized；
Step 3：The culture of explant, the culture medium of inoculation explant is placed into culturing room and cultivated 1 month, so as to obtain nothing Vaccine, condition of culture are to cultivate 25 DEG C of room temperature, the lux of intensity of illumination 1200, daily light application time 10h；
Step 4：Subinoculation and culture, the seedling will be sprouted on transfer room superclean bench, be individually inoculated into the present invention In subculture root media on, cultivate 2 months, cultivate 25 DEG C of room temperature, the lux of intensity of illumination 1200, daily light application time 10h。
Step 5：Repeat the above steps four, after 2 squamous subculture cultures, take root good and have a larger protocorm, Directly it can be transplanted in matrix, normal maintenance obtains finished product seedling.
Comparative example is tested
Comparative example 1：Composite reagent is configured first, includes mother liquor I, mother liquor II, mother liquor III and mother liquor IV,
Wherein, mother liquor I：KNO3 38g、NH4NO3 33g、CaCl2·2H2O 8.8g、MgSO4·7H2O 7.4g、KH2PO4 3.4g；
Mother liquor II：KI 0.166g、H3BO3 1.24g、MnSO4·4H2O 4.46g、ZnSO4·7H2O 1.72g、Na2MoO4· 2H2O 0.05g、CuSO4·5H2O 0.005g、CoCl2·6H2O 0.005g；
Mother liquor III：Inositol 20g, nicotinic acid 0.1g, pyridoxine hydrochloride 0.1g, thiamine hydrochloride 0.02g, glycine 0.4g；
Growth promoter, include 6-BA and IAA that concentration is 500mg/L；
Nutritious supplementary pharmaceutical, every liter of nutritious supplementary pharmaceutical include pyrolkigneous liquid 60mL, tomato juice 240mL, ginkgo liquid extraction liquid 75ml, sucrose 105g, agar powder 15g.
Bletilla striata subculture medium and making：
S1：The mother liquor I as in above-mentioned composite reagent prefabricated first, mother liquor II, mother liquor III, growth promoter, nutritious supplementary pharmaceutical, And prepare to spend precious and agar powder to be refrigerated；
S2：The 50ml of mother liquor I, mother liquor II, mother liquor III, 5ml, nutritious supplementary pharmaceutical 300ml and life are chosen from prefabricated composite reagent Long accelerator 0.08ml, precious 3.5g is spent, agar powder 6.5g, various solution are sufficiently mixed to obtain mixed liquor A, then using dense The NaOH or HCl that spend for 0.5mol/L adjust the pH value of mixed liquor A, end when ph values reach 5.8；
S3：It is dispensed into every bottle of 80ML/ blake bottle, is put into high-pressure sterilizing pot, sterilizes 25min, high-pressure sterilizing pot work temperature Degree 120 DEG C, pressure 120kPa, after the completion of refrigeration place it is stand-by.
Comparative example 2：Composite reagent is configured first, includes mother liquor I, mother liquor II, mother liquor III and mother liquor IV,
Wherein, mother liquor I：KNO3 38g、NH4NO3 33g、CaCl2·2H2O 8.8g、MgSO4·7H2O 7.4g、KH2PO4 3.4g；
Mother liquor II：KI 0.166g、H3BO3 1.24g、MnSO4·4H2O 4.46g、ZnSO4·7H2O 1.72g、Na2MoO4· 2H2O 0.05g、CuSO4·5H2O 0.005g、CoCl2·6H2O 0.005g；
Mother liquor III：Inositol 20g, nicotinic acid 0.1g, pyridoxine hydrochloride 0.1g, thiamine hydrochloride 0.02g, glycine 0.4g；
Mother liquor IV：FeSO4·7H2O 5.56g、Na2·EDTA·2H2O 7.46g；
Nutritious supplementary pharmaceutical, every liter of nutritious supplementary pharmaceutical include pyrolkigneous liquid 60mL, tomato juice 240mL, ginkgo liquid extraction liquid 75ml, sucrose 105g, agar powder 15g.
Bletilla striata subculture medium and making：
S1：The mother liquor I as in above-mentioned composite reagent prefabricated first, mother liquor II, mother liquor III and mother liquor IV, growth promoter, battalion Replenishers are supported, and prepare to spend precious and agar powder to be refrigerated；
S2：The 50ml of mother liquor I, mother liquor II, mother liquor III, the 5ml of mother liquor IV, nutritious supplementary pharmaceutical are chosen from prefabricated composite reagent 300ml, precious 3.5g, agar powder 6.5g are spent, various solution are sufficiently mixed to obtain mixed liquor A, and then concentration is The pH value of 0.5mol/L NaOH or HCl regulation mixed liquor A, ends when ph values reach 5.8；
S3：It is dispensed into every bottle of 80ML/ blake bottle, is put into high-pressure sterilizing pot, sterilizes 25min, high-pressure sterilizing pot work temperature Degree 120 DEG C, pressure 120kPa, after the completion of refrigeration place it is stand-by.
Comparative example 3：Composite reagent is configured first, includes mother liquor I, mother liquor II, mother liquor III and mother liquor IV,
Wherein, mother liquor I：KNO3 38g、NH4NO3 33g、CaCl2·2H2O 8.8g、MgSO4·7H2O 7.4g、KH2PO4 3.4g；
Mother liquor II：KI 0.166g、H3BO3 1.24g、MnSO4·4H2O 4.46g、ZnSO4·7H2O 1.72g、Na2MoO4· 2H2O 0.05g、CuSO4·5H2O 0.005g、CoCl2·6H2O 0.005g；
Mother liquor III：Inositol 20g, nicotinic acid 0.1g, pyridoxine hydrochloride 0.1g, thiamine hydrochloride 0.02g, glycine 0.4g；
Mother liquor IV：FeSO4·7H2O 5.56g、Na2·EDTA·2H2O 7.46g；
Growth promoter, include 6-BA and IAA that concentration is 500mg/L.
Bletilla striata subculture medium and making：
S1：The mother liquor I as in above-mentioned composite reagent prefabricated first, mother liquor II, mother liquor III and mother liquor IV, growth promoter, and Preparation spends precious and agar powder to be refrigerated；
S2：The 50ml of mother liquor I, mother liquor II, mother liquor III, the 5ml of mother liquor IV, and growth promoter are chosen from prefabricated composite reagent 0.08ml, precious 3.5g, agar powder 6.5g are spent, various solution are sufficiently mixed to obtain mixed liquor A, and then concentration is The pH value of 0.5mol/L NaOH or HCl regulation mixed liquor A, ends when ph values reach 5.8；
S3：It is dispensed into every bottle of 80ML/ blake bottle, is put into high-pressure sterilizing pot, sterilizes 25min, high-pressure sterilizing pot work temperature Degree 120 DEG C, pressure 120kPa, after the completion of refrigeration place it is stand-by.
Comparative example 4：Composite reagent is configured first, includes mother liquor I, mother liquor II, mother liquor III and mother liquor IV,
Wherein, mother liquor I：KNO3 38g、NH4NO3 33g、CaCl2·2H2O 8.8g、MgSO4·7H2O 7.4g、KH2PO4 3.4g；
Mother liquor II：KI 0.166g、H3BO3 1.24g、MnSO4·4H2O 4.46g、ZnSO4·7H2O 1.72g、Na2MoO4· 2H2O 0.05g、CuSO4·5H2O 0.005g、CoCl2·6H2O 0.005g；
Mother liquor IV：FeSO4·7H2O 5.56g、Na2·EDTA·2H2O 7.46g；
Growth promoter, include 6-BA and IAA that concentration is 500mg/L；
Nutritious supplementary pharmaceutical, every liter of nutritious supplementary pharmaceutical include pyrolkigneous liquid 60mL, tomato juice 240mL, ginkgo liquid extraction liquid 75ml, sucrose 105g, agar powder 15g.
Bletilla striata subculture medium and making：
S1：The mother liquor I as in above-mentioned composite reagent prefabricated first, mother liquor II and mother liquor IV, growth promoter, nutritional supplementation Agent, and prepare to spend precious and agar powder to be refrigerated；
S2：The 50ml of mother liquor I, mother liquor II, mother liquor IV 5ml, nutritious supplementary pharmaceutical 300ml and growth are chosen from prefabricated composite reagent Accelerator 0.08ml, precious 3.5g, agar powder 6.5g are spent, various solution are sufficiently mixed to obtain mixed liquor A, then concentration The pH value of mixed liquor A is adjusted for 0.5mol/L NaOH or HCl, is ended when ph values reach 5.8；
S3：It is dispensed into every bottle of 80ML/ blake bottle, is put into high-pressure sterilizing pot, sterilizes 25min, high-pressure sterilizing pot work temperature Degree 120 DEG C, pressure 120kPa, after the completion of refrigeration place it is stand-by.
In squamous subculture, the size of bletilla striata seedling weightening weight can fully demonstrate the upgrowth situation of the bletilla striata, weightening weight The big then bletilla striata growing way of amount is better, and detection process is chosen according to the deployed each 100ml of culture medium of component in embodiment and comparative example, 10 plants are weighed and the bletilla striata seedling of similar weight is put into every kind of subculture medium, 15 after culture day and progress in 30 days Weighing measurement.
The unit for detecting the weightening weight in data is g（FW）/ml-1。
Embodiment 1 Embodiment 2 Embodiment 3 Embodiment 4 Embodiment 5 Comparative example 1 Comparative example 2 Comparative example 3 Comparative example 4
15d weights 0.1 0.08 0.07 0.1 0.08 0.05 0.05 0.04 0.04
30d weights 0.62 0.58 0.60 0.64 0.61 0.34 0.37 0.31 0.51
By the contrast between comparative example and embodiment it can be found that having used the parameter in embodiment example 1-5 to be adjusted Culture medium can be very good promote the bletilla striata weightening, so as to ensure smoothly completing for squamous subculture, and lack in comparative example Necessary component, although so as to which subculture can also smoothly complete, growing way has a certain distance relative to embodiment really, is not A kind of preferably plant generation state.
This specific embodiment is only explanation of the invention, and it is not limitation of the present invention, people in the art Member can make the modification of no creative contribution to the present embodiment as needed after this specification is read, but as long as at this All protected in the right of invention by Patent Law.

Claims (8)

  1. A kind of 1. composite reagent, it is characterised in that：Include mother liquor I, mother liquor II, mother liquor III and mother liquor IV,
    Wherein, every liter of mother liquor I：KNO3 30-46g、NH4NO3 27-39g、CaCl2·2H2O 8-9.6g、MgSO4·7H2O 7- 7.8g、KH2PO43-3.8g；
    Every liter of mother liquor II：KI 0.16-0.172g、H3BO3 1.2-1.28g、MnSO4·4H2O 4.4-4.52g、ZnSO4·7H2O 1.7-1.74g、Na2MoO4·2H2O 0.03-0.07g、CuSO4·5H2O 0.03-0.07g、CoCl2·6H2O 0.03- 0.07g；
    Every liter of mother liquor III：Inositol 16-24g, nicotinic acid 0.07-0.13g, pyridoxine hydrochloride 0.07-0.13g, thiamine hydrochloride 0.01-0.03g, glycine 0.2-0.6g；
    Every liter of mother liquor IV：FeSO4·7H2O 5.5-5.62g、Na2·EDTA·2H2O 7.46g。
  2. A kind of 2. composite reagent according to claim 1, it is characterised in that：Also include what is promoted plant growth and divide Growth promoter.
  3. A kind of 3. composite reagent according to claim 2, it is characterised in that：Growth promoter includes 6-BA and IAA two Plant Hormone.
  4. A kind of 4. composite reagent according to claim 3, it is characterised in that：6-BA and IAA weight ratio is 1:1, wherein 6-BA and IAA concentration is 500mg/L.
  5. A kind of 5. composite reagent according to claim 1, it is characterised in that：Also include nutritious supplementary pharmaceutical, every liter of nutrition Replenishers include pyrolkigneous liquid 60mL, tomato juice 240mL, ginkgo liquid extraction liquid 75ml, sucrose 105g, agar powder 15g.
  6. A kind of 6. application of composite reagent in bletilla striata incubation.
  7. A kind of 7. bletilla striata subculture medium, it is characterised in that：Include the 45-55ml of mother liquor I, mother liquor II, mother in every liter of culture medium Liquid III and the 3-7ml of mother liquor IV, growth promoter 0.06-0.1ml, nutritious supplementary pharmaceutical 200-400ml, spend precious 2.5-4.5g, agar Powder 5.5-7.5g.
  8. A kind of 8. preparation method of bletilla striata subculture medium, it is characterised in that：Comprise the following steps：
    S1：Mother liquor I, mother liquor II, mother liquor III, mother liquor IV and nutritious supplementary pharmaceutical and each some liters of growth promoter are prepared, and are deposited It is stored in refrigerating chamber；
    S2：When needing to use culture medium, according to proportioning by mother liquor I, mother liquor II, mother liquor III, mother liquor IV and nutritious supplementary pharmaceutical It is sufficiently mixed to obtain mixed liquor A with growth promoter, NaOH or the HCl regulation that then concentration is 0.5mol/L are mixed Liquid A pH value is closed, is ended when ph values reach 5.8；
    S3：It is dispensed into every bottle of 80ML/ blake bottle, is put into high-pressure sterilizing pot, sterilizes 25min, high-pressure sterilizing pot work temperature Degree 120 DEG C, pressure 120kPa, after the completion of refrigeration place it is stand-by.
CN201710836183.4A 2017-09-16 2017-09-16 A kind of composite reagent and its application in the bletilla striata is cultivated Pending CN107568062A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112369326A (en) * 2020-11-05 2021-02-19 北京花乡花木集团有限公司 Tissue culture method of hedyotis diffusa' Hadesben ice cream

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112369326A (en) * 2020-11-05 2021-02-19 北京花乡花木集团有限公司 Tissue culture method of hedyotis diffusa' Hadesben ice cream

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Application publication date: 20180112