CN102884981B - Zanthoxylum nitidum tissue culture medium - Google Patents

Abstract

The invention discloses a zanthoxylum nitidum tissue culture medium. The medium comprises a subculture medium and a rooting medium. Improvement is carried out on the basis of an H medium, the macroelement usage is adjusted, and the iron vitriol content is increased. The medium is suitable for culturing zanthoxylum nitidum and single zanthoxylum nitidum tissues from different producing areas. The subculture period takes 20-25 days, the growth coefficient is 4-6, subculture seedlings grow well, small seedlings are thick and order, the leaf color is green, no etiolated seedlings are produced, and the seedling yield is high. The rooting rate of the rooted seedlings can reach 90%, the rooting is quick; and when the temperature is at 25 plus or minus 2 DEG C, the average rooting time is 10-15 days, a root system well grows, the rooting number is larger and averagely 3-5 roots, the seedling is good in lignification, the transplanting survival rate reaches above 90%, and the outplanting time of bottled seedlings is about 30 days averagely.

Description

The Shinyleaf Pricklyash Root culture medium for tissue culture

Technical field

The present invention relates to tissue culture medium (TCM), relate in particular to a kind of Shinyleaf Pricklyash Root culture medium for tissue culture.

Background technology

Shinyleaf Pricklyash Root (Zanthoxylum nitidum), be rutaceae, because of all there is little hook thorn on its blade Zhong Mai two sides, so claim " Shinyleaf Pricklyash Root ", have another name called two dorsal stylets, nail-plate thorn (Foochow), go under mountain tiger, anaesthetic rattan, Radix Zanthoxyli, the leaf to thread a needle, red barb muscle, Da Ye retouch pawl le (Guangdong, Guangxi).Shinyleaf Pricklyash Root happiness is grown in warm, sun-drenched place, in the sparse woods on mountain region, hills, level land, the shrubbery, grass slope, deserted mountain have in the thornbush more commonly, mainly be distributed in ground such as China Guangdong, Guangxi, Fujian, Hainan, Yunnan, Guizhou and Taiwan.Shinyleaf Pricklyash Root is China's a kind of Chinese medicinal materials commonly used, and rhizome, leaf, pericarp all can be used as medicine, the effects such as the stasis of blood, analgesia, detumescence of invigorating blood circulation, loose.Modern medicinal chemical research shows: the Shinyleaf Pricklyash Root rhizome contains alkaloid oxynitidine, N-demethoxychelerythrone, dihydronitidine, skimmianine, 6-ethoxychelerythrine, α-allocryptopine, nitidine etc.During topical application, nerve ending is had anesthetic action, and anaesthetic effect is stable, has no adverse reaction, and does not also have hepatorenal damage etc., is a kind of Chinese medicinal materials with wide prospect in medicine, therefore carries out the fundamental research of Shinyleaf Pricklyash Root, has crucial meaning.

At present, domestic more existing Shinyleaf Pricklyash Root tissue culture technique preliminary study reports, the general combination of adopting minimum medium (MS) and growth regulator in the selection of substratum, but Shinyleaf Pricklyash Root provenance place of production difference, conditions of tissue culture is also different, even the Shinyleaf Pricklyash Root provenance in the same place of production selects for use different individual plants to make explant, conditions of tissue culture is also different.The MS substratum is applicable to part place of production Shinyleaf Pricklyash Root tissue culture, and part place of production Shinyleaf Pricklyash Root provenance and Shinyleaf Pricklyash Root individual plant carry out succeeding transfer culture with the MS substratum, first blade flavescence, bleaches, and stem flavescence then, bleaches, and the death of bud seedling can't be proceeded succeeding transfer culture.If different places of production Shinyleaf Pricklyash Root provenance or Shinyleaf Pricklyash Root individual plant use a kind of substratum, the production operation difficulty is big, and the cost height is unfavorable for large-scale production.

Summary of the invention

The objective of the invention is to overcome the deficiencies in the prior art, a kind of Shinyleaf Pricklyash Root culture medium for tissue culture is provided, this substratum comprises subculture medium and root media, wherein:

Component and each components contents of every liter of (L) subculture medium are as follows:

(1) macroelement: saltpetre 1250mg/L, ammonium nitrate 1000mg/L, magnesium sulfate heptahydrate 370mg/L, Calcium dichloride dihydrate 166mg/L, potassium primary phosphate 150mg/L, disodium ethylene diamine tetraacetate 56.0mg/L, iron vitriol 41.7mg/L;

(2) trace element: four water manganous sulfate 25mg/L, Zinc Sulphate Heptahydrate 10mg/L, boric acid 10mg/L, Sodium Molybdate Dihydrate 0.25mg/L, potassiumiodide 0.083mg/L, cupric sulfate pentahydrate 0.025mg/L;

(3) organism: inositol 100.0mg/L, glycine 2.0mg/L, vitamin (VB ₁) 0.5mg/L, pyridoxine hydrochloride (VB ₆) 0.5mg/L, folic acid 0.5mg/L, vitamin H (VH) 0.05mg/L;

(4) plant-growth regulator: 6-benzyladenine 0.4～1.0mg/L, naphthylacetic acid 0.2～0.5mg/L;

Surplus is a distilled water.

Component and each components contents of every liter of (L) root media are as follows:

(1) macroelement: saltpetre 1250mg/L, ammonium nitrate 1000mg/L, magnesium sulfate heptahydrate 370mg/L, Calcium dichloride dihydrate 166mg/L, potassium primary phosphate 150mg/L, iron vitriol 33.4mg/L, disodium ethylene diamine tetraacetate 44.8mg/L;

(2) trace element: four water manganous sulfate 25mg/L, Zinc Sulphate Heptahydrate 10mg/L, boric acid 10mg/L, Sodium Molybdate Dihydrate 0.25mg/L, potassiumiodide 0.083mg/L, cupric sulfate pentahydrate 0.025mg/L;

(3) organism: inositol 100.0mg/L, glycine 2.0mg/L, vitamin (VB ₁) 0.5mg/L, pyridoxine hydrochloride (VB ₆) 0.5mg/L, folic acid 0.5mg/L, vitamin H (VH) 0.05mg/L;

(4) plant-growth regulator: root-inducing powder ABT6 number 0.4～0.8mg/L, naphthylacetic acid 0.2～0.4mg/L;

Surplus is a distilled water.

Subculture in the Shinyleaf Pricklyash Root culture medium for tissue culture of the present invention, the compound method of root media are:

The preparation of 1 mother liquor

1.1 for ease of sampling, the various mother liquors of suitable preparation earlier are divided into macroelement mother liquor, micro-mother liquor, organism mother liquor and each plant growth regulators mother liquor.Sucrose, agar should not be made into mother liquor, directly claim sample when needing;

1.2 each concentration of component of macroelement mother liquor and organism mother liquor should become 10～100 multiple relation with the concentration of component of substratum, micro-mother liquor becomes 200～500 multiple relation, and the concentration of plant growth regulating reagent mother liquor should be made into 1mg/ml;

1.3 mother liquor is selected aseptic distilled water, deionized water or ultrapure water preparation for use, uses the water that boiled during mass production;

1.4 during preparation macroelement mother liquor, each component should be dissolved separately, mix one by one by the order of nitrogen, calcium, magnesium, phosphorus the back, otherwise cause precipitation easily;

1.5 micro-mother liquor contains the ferrous sulfate of seeing labile potassiumiodide of light and easy oxidation, uses brown bottle and preserves;

1.6 plant-growth regulator mother liquor and organism mother liquor should place refrigerator to preserve;

1.7 should in time use after the mother liquor preparation, period of storage should not be above 1 month;

1.8 find that mother liquor has precipitation, or microorganism growth arranged, or algal grown is arranged, should pass into disuse.

The preparation of 2 substratum

2.1 according to culture medium prescription, measure mother liquor in proportion, go in the containers such as Stainless Steel Kettle after adding the water constant volume;

2.2 add an amount of peptizer and sugar, and stir peptizer is fully disperseed, sugar fully dissolves;

2.3 be heated to firm boiling, peptizer dissolved fully.When if the self-reacting device that adopts the limit to divide rim to stir is prepared substratum, can save heating and make peptizer dissolved process;

2.4 be adjusted to suitable potential of hydrogen with 1.0N hydrochloric acid or 1.0N sodium hydroxide;

2.5 as early as possible the substratum branch is installed in the incubator ware, in order to avoid culture medium solidifying or retrogradation and be difficult to packing;

2.6 should avoid substratum to adhere to bottleneck during the packing substratum,, should wipe totally, otherwise cause pollution easily in case be stained with.

The sterilization of 3 substratum

3.1 the substratum that branch is installed is put into autoclave sterilizer and is sterilized;

3.2 the heating initial stage when the air pressure of sterilizing pan sterilized room reaches 0.05Mpa, is opened condensation trap, drains freezing air in the sterilized room;

3.3, keep temperature and pressure sterilization 15～20 minutes when the sterilized room internal gas pressure reaches 0.11Mpa, when temperature reaches 121 ℃, picks up counting;

3.4 close the heating power supply switch, by slow exhaust mode discharging hot gas, when reducing to normal atmosphere, opens indoor air pressure to be sterilized sterilization pot cover or door, take out substratum;

3.5 substratum should place little air to flow and the clean environment cooling of few dust, otherwise causes mould spores to enter the cultivation vessel in the cooling intake process, produces mould contamination.Or the thick cloth parcel cooling of sterilizing with high pressure-temperature.

4 substratum storage

4.1 substratum should be now with the current as far as possible.

4.2 can be at air clean and immobilising environment short period of time storage substratum, should pass into disuse but storage surpasses 1 month substratum.

The method of Shinyleaf Pricklyash Root tissue culture of the present invention is:

1. bud induces

Choose Shinyleaf Pricklyash Root plant robust growth, no disease and pest in fine day, the young shoot of clip semi-lignified is as explant, cut off blade (staying petiole), clean 1～2 time with the water that is added with a little liquid detergent, rinse well with flowing water again, then on Bechtop with 0.1% mercury chloride sterilization 6-7 minute and shake for several times, use sterile water wash at last 4～5 times, the material that has disinfected is sheared the long section into about 2cm, every section is stayed 1～2 axillalry bud, then oblique cutting or vertically be inserted in the substratum of induced bud.When explant is cultivated for the first time, need 10～15 days axillalry buds of complete dark cultivation can begin to sprout, and then move to the place cultivation of seeing light.

2. the multiplication culture of bud

Sprout when growing to the 1cm left and right sides when axillalry bud, cut axillalry bud and induce the differentiation bud of growing thickly can carry out the shoot proliferation of bud, general one month subculture once, the subculture material all is placed on indoor cultivation, culturing room's temperature is (26 ± 2) ℃, illumination 12h.d ^-1, illuminance 2000～3000lx.

3. what take root induces

When the bud of growing thickly grows to the 3cm left and right sides, can choose healthy and strong bud and transfer in the root media, carry out root induction, all the other bud clumps can be transferred to and allow it continue propagation in the subculture medium of newly joining.After budling changes in the root media, cultivating about 15 days under the indoor weak light condition earlier, the root of hair rate is moved outdoor hardening booth in natural scattering illumination cultivation 10～20 day (autumn and winter time slightly long) to (the low indoor cultivation of winter temperature is about 20 days, and the root of hair rate reaches more than 70%) at 50% o'clock.

4. the take root transplanting of seedling

The abundant lignifying of seedling of waiting to take root, blade is unfolded, and the leaf look dark green, and the seedling stem is dark green, the caulom elongation, height of seedling 4cm can transplant during the left and right sides.Pour out substratum during transplanting, take out seedling, wash the substratum that remains on the root, and clip long root system, it is long to keep 2～2.5cm, transplants in the nutrient bag of sterilizing through 0.2% potassium permanganate, the water of drenching, coating film keeps certain temperature and humidity, and will avoid the sunlight direct projection.

Advantage of the present invention is:

1, the Shinyleaf Pricklyash Root provenance place of production is different with individual plant, and conditions of tissue culture is also different, and the MS substratum is applicable to part place of production Shinyleaf Pricklyash Root tissue culture, and the present invention is more suitable for the Shinyleaf Pricklyash Root in the different places of production and the tissue culture of different Shinyleaf Pricklyash Root individual plants.

2, part place of production Shinyleaf Pricklyash Root provenance is carried out succeeding transfer culture with the MS substratum, first blade flavescence, bleaches, and stem flavescence then, bleaches, and can't proceed succeeding transfer culture; With substratum of the present invention Shinyleaf Pricklyash Root is cultivated, adjusted the macroelement consumption, increased the content of iron vitriol, subculture cycle 20～25 days, growth coefficient 4～6, subculture seedling well-grown, seedling is sturdy, neat, the leaf look dark green, etiolated seedling (flavescence gradually of blade, cauline leaf) few, gets seedling rate height.

3, in root media, increase the content of iron vitriol, cultivate with root media that the seedling rooting rate of taking root can reach 90%, to go out root fast, during 25 ± 2 ℃ of temperature, on average going out the root time is 10～15 days, well developed root system, and how go out the root amount on average has 3～5 roots, the seedling lignifying is good, transplanting survival rate height, transplanting survival rate reach more than 90%, and it is about 30 days that the bottle seedling goes out the garden time average.

Compare with the Shinyleaf Pricklyash Root tissue culture medium (TCM) of present prior art, advantage of the present invention is:

Embodiment

The invention will be further described with embodiment below, but the present invention is not limited to these embodiment.

Embodiment 1:

The Shinyleaf Pricklyash Root culture medium for tissue culture, this substratum comprises subculture medium and root media, wherein:

Component and each components contents of every liter of (L) subculture medium are as follows:

(1) macroelement: saltpetre 1250mg/L, ammonium nitrate 1000mg/L, magnesium sulfate heptahydrate 370mg/L, Calcium dichloride dihydrate 166mg/L, potassium primary phosphate 150mg/L, disodium ethylene diamine tetraacetate 56.0mg/L, iron vitriol 41.7mg/L;

(2) trace element: four water manganous sulfate 25mg/L, Zinc Sulphate Heptahydrate 10mg/L, boric acid 10mg/L, Sodium Molybdate Dihydrate 0.25mg/L, potassiumiodide 0.083mg/L, cupric sulfate pentahydrate 0.025mg/L;

(3) organism: inositol 100.0mg/L, glycine 2.0mg/L, vitamin (VB ₁) 0.5mg/L, pyridoxine hydrochloride (VB ₆) 0.5mg/L, folic acid 0.5mg/L, vitamin H (VH) 0.05mg/L;

(4) plant-growth regulator: 6-benzyladenine 0.4mg/L, naphthylacetic acid 0.2mg/L;

Surplus is a distilled water.

Component and each components contents of every liter of (L) root media are as follows:

(1) macroelement: saltpetre 1250mg/L, ammonium nitrate 1000mg/L, magnesium sulfate heptahydrate 370mg/L, Calcium dichloride dihydrate 166mg/L, potassium primary phosphate 150mg/L, iron vitriol 33.4mg/L, disodium ethylene diamine tetraacetate 44.8mg/L;

(2) trace element: four water manganous sulfate 25mg/L, Zinc Sulphate Heptahydrate 10mg/L, boric acid 10mg/L, Sodium Molybdate Dihydrate 0.25mg/L, potassiumiodide 0.083mg/L, cupric sulfate pentahydrate 0.025mg/L;

(3) organism: inositol 100.0mg/L, glycine 2.0mg/L, vitamin (VB ₁) 0.5mg/L, pyridoxine hydrochloride (VB ₆) 0.5mg/L, folic acid 0.5mg/L, vitamin H (VH) 0.05mg/L;

(4) plant-growth regulator: root-inducing powder ABT6 0.4mg/L, naphthylacetic acid 0.2mg/L;

Surplus is a distilled water.

Embodiment 2:

The Shinyleaf Pricklyash Root culture medium for tissue culture, this substratum comprises subculture medium and root media, wherein:

Component and each components contents of every liter of (L) subculture medium are as follows:

(1) macroelement: saltpetre 1250mg/L, ammonium nitrate 1000mg/L, magnesium sulfate heptahydrate 370mg/L, Calcium dichloride dihydrate 166mg/L, potassium primary phosphate 150mg/L, disodium ethylene diamine tetraacetate 56.0mg/L, iron vitriol 41.7mg/L;

(2) trace element: four water manganous sulfate 25mg/L, Zinc Sulphate Heptahydrate 10mg/L, boric acid 10mg/L, Sodium Molybdate Dihydrate 0.25mg/L, potassiumiodide 0.083mg/L, cupric sulfate pentahydrate 0.025mg/L;

(3) organism: inositol 100.0mg/L, glycine 2.0mg/L, vitamin (VB ₁) 0.5mg/L, pyridoxine hydrochloride (VB ₆) 0.5mg/L, folic acid 0.5mg/L, vitamin H (VH) 0.05mg/L;

(4) plant-growth regulator: 6-benzyladenine 1.0mg/L, naphthylacetic acid 0.5mg/L;

Surplus is a distilled water.

Component and each components contents of every liter of (L) root media are as follows:

(1) macroelement: saltpetre 1250mg/L, ammonium nitrate 1000mg/L, magnesium sulfate heptahydrate 370mg/L, Calcium dichloride dihydrate 166mg/L, potassium primary phosphate 150mg/L, iron vitriol 33.4mg/L, disodium ethylene diamine tetraacetate 44.8mg/L;

(2) trace element: four water manganous sulfate 25mg/L, Zinc Sulphate Heptahydrate 10mg/L, boric acid 10mg/L, Sodium Molybdate Dihydrate 0.25mg/L, potassiumiodide 0.083mg/L, cupric sulfate pentahydrate 0.025mg/L;

(3) organism: inositol 100.0mg/L, glycine 2.0mg/L, vitamin (VB ₁) 0.5mg/L, pyridoxine hydrochloride (VB ₆) 0.5mg/L, folic acid 0.5mg/L, vitamin H (VH) 0.05mg/L;

(4) plant-growth regulator: root-inducing powder ABT6 0.8mg/L, naphthylacetic acid 0.4mg/L;

Surplus is a distilled water.

Embodiment 3:

The Shinyleaf Pricklyash Root culture medium for tissue culture, this substratum comprises subculture medium and root media, wherein:

Component and each components contents of every liter of (L) subculture medium are as follows:

(1) macroelement: saltpetre 1250mg/L, ammonium nitrate 1000mg/L, magnesium sulfate heptahydrate 370mg/L, Calcium dichloride dihydrate 166mg/L, potassium primary phosphate 150mg/L, disodium ethylene diamine tetraacetate 56.0mg/L, iron vitriol 41.7mg/L;

(2) trace element: four water manganous sulfate 25mg/L, Zinc Sulphate Heptahydrate 10mg/L, boric acid 10mg/L, Sodium Molybdate Dihydrate 0.25mg/L, potassiumiodide 0.083mg/L, cupric sulfate pentahydrate 0.025mg/L;

(3) organism: inositol 100.0mg/L, glycine 2.0mg/L, vitamin (VB ₁) 0.5mg/L, pyridoxine hydrochloride (VB ₆) 0.5mg/L, folic acid 0.5mg/L, vitamin H (VH) 0.05mg/L;

(4) plant-growth regulator: 6-benzyladenine 0.6mg/L, naphthylacetic acid 0.3mg/L;

Surplus is a distilled water.

Component and each components contents of every liter of (L) root media are as follows:

(1) macroelement: saltpetre 1250mg/L, ammonium nitrate 1000mg/L, magnesium sulfate heptahydrate 370mg/L, Calcium dichloride dihydrate 166mg/L, potassium primary phosphate 150mg/L, iron vitriol 33.4mg/L, disodium ethylene diamine tetraacetate 44.8mg/L;

(2) trace element: four water manganous sulfate 25mg/L, Zinc Sulphate Heptahydrate 10mg/L, boric acid 10mg/L, Sodium Molybdate Dihydrate 0.25mg/L, potassiumiodide 0.083mg/L, cupric sulfate pentahydrate 0.025mg/L;

(3) organism: inositol 100.0mg/L, glycine 2.0mg/L, vitamin (VB ₁) 0.5mg/L, pyridoxine hydrochloride (VB ₆) 0.5mg/L, folic acid 0.5mg/L, vitamin H (VH) 0.05mg/L;

(4) plant-growth regulator: root-inducing powder ABT6 0.6mg/L, naphthylacetic acid 0.3mg/L;

Surplus is a distilled water.

Embodiment 4:

The Shinyleaf Pricklyash Root culture medium for tissue culture, this substratum comprises subculture medium and root media, wherein:

Component and each components contents of every liter of (L) subculture medium are as follows:

(1) macroelement: saltpetre 1250mg/L, ammonium nitrate 1000mg/L, magnesium sulfate heptahydrate 370mg/L, Calcium dichloride dihydrate 166mg/L, potassium primary phosphate 150mg/L, disodium ethylene diamine tetraacetate 56.0mg/L, iron vitriol 41.7mg/L;

(2) trace element: four water manganous sulfate 25mg/L, Zinc Sulphate Heptahydrate 10mg/L, boric acid 10mg/L, Sodium Molybdate Dihydrate 0.25mg/L, potassiumiodide 0.083mg/L, cupric sulfate pentahydrate 0.025mg/L;

(3) organism: inositol 100.0mg/L, glycine 2.0mg/L, vitamin (VB ₁) 0.5mg/L, pyridoxine hydrochloride (VB ₆) 0.5mg/L, folic acid 0.5mg/L, vitamin H (VH) 0.05mg/L;

(4) plant-growth regulator: 6-benzyladenine 1.0mg/L, naphthylacetic acid 0.2mg/L;

Surplus is a distilled water.

Component and each components contents of every liter of (L) root media are as follows:

(1) macroelement: saltpetre 1250mg/L, ammonium nitrate 1000mg/L, magnesium sulfate heptahydrate 370mg/L, Calcium dichloride dihydrate 166mg/L, potassium primary phosphate 150mg/L, iron vitriol 33.4mg/L, disodium ethylene diamine tetraacetate 44.8mg/L;

(2) trace element: four water manganous sulfate 25mg/L, Zinc Sulphate Heptahydrate 10mg/L, boric acid 10mg/L, Sodium Molybdate Dihydrate 0.25mg/L, potassiumiodide 0.083mg/L, cupric sulfate pentahydrate 0.025mg/L;

(3) organism: inositol 100.0mg/L, glycine 2.0mg/L, vitamin (VB ₁) 0.5mg/L, pyridoxine hydrochloride (VB ₆) 0.5mg/L, folic acid 0.5mg/L, vitamin H (VH) 0.05mg/L;

(4) plant-growth regulator: root-inducing powder ABT6 0.6mg/L, naphthylacetic acid 0.4mg/L;

Surplus is a distilled water.

Comparative Examples:

Subculture medium MS+6-BA2.0+NAA1.0, root media 1/2MS+ABT ₁0.8+IBA0.6 concentration unit is mg/L.

The substratum that utilizes the foregoing description 1 and Comparative Examples to make carries out tissue culture to Shinyleaf Pricklyash Root, and from the result of following table as can be seen, the technical indicator of embodiment far surpasses Comparative Examples.

Claims (1)

1. Shinyleaf Pricklyash Root culture medium for tissue culture, this substratum comprises subculture medium and root media, it is characterized in that:

Component and each components contents of every liter of (L) subculture medium are as follows:

(1) macroelement: saltpetre 1250mg/L, ammonium nitrate 1000mg/L, magnesium sulfate heptahydrate 370mg/L, Calcium dichloride dihydrate 166mg/L, potassium primary phosphate 150mg/L, disodium ethylene diamine tetraacetate 56.0mg/L, iron vitriol 41.7mg/L;

(2) trace element: four water manganous sulfate 25mg/L, Zinc Sulphate Heptahydrate 10mg/L, boric acid 10mg/L, Sodium Molybdate Dihydrate 0.25mg/L, potassiumiodide 0.083mg/L, cupric sulfate pentahydrate 0.025mg/L;

(3) organism: inositol 100.0mg/L, glycine 2.0mg/L, vitamin 0.5mg/L, pyridoxine hydrochloride 0.5mg/L, folic acid 0.5mg/L, vitamin H 0.05mg/L;

(4) plant-growth regulator: 6-benzyladenine 0.4-1.0mg/L, naphthylacetic acid 0.2-0.5mg/L;

Surplus is a distilled water;

Component and each components contents of every liter of (L) root media are as follows:

(1) macroelement: saltpetre 1250mg/L, ammonium nitrate 1000mg/L, magnesium sulfate heptahydrate 370mg/L, Calcium dichloride dihydrate 166mg/L, potassium primary phosphate 150mg/L, iron vitriol 33.4mg/L, disodium ethylene diamine tetraacetate 44.8mg/L;

(2) trace element: four water manganous sulfate 25mg/L, Zinc Sulphate Heptahydrate 10mg/L, boric acid 10mg/L, Sodium Molybdate Dihydrate 0.25mg/L, potassiumiodide 0.083mg/L, cupric sulfate pentahydrate 0.025mg/L;

(3) organism: inositol 100.0mg/L, glycine 2.0mg/L, vitamin 0.5mg/L, pyridoxine hydrochloride 0.5mg/L, folic acid 0.5mg/L, vitamin H 0.05mg/L;

(4) plant-growth regulator: root-inducing powder ABT6 0.4-0.8mg/L, naphthylacetic acid 0.2-0.4mg/L;

Surplus is a distilled water.