CN104705193B - A kind of japonica rice flower pesticide Fiber differentiation based formulas - Google Patents

Abstract

The invention provides a kind of culture medium for the japonica rice flower pesticide Fiber differentiation stage, the formula of this culture medium is by a certain proportion of KNO₃、NH₄NO₃、Ca(NO₃)₂∙4H₂0、KH₂PO₄、KCl、MgSO₄∙7H₂O、Na₂‑EDTA、FeSO₄∙7H₂O、MnSO₄∙4H₂O、ZnSO₄∙7H₂O、H₃BO₃, KI, inositol, glycerine, Sodium Citrate, usp, Dihydrate Powder, folic acid, glycine, lysine, vitamin B1, vitamin B6, nicotinic acid, sucrose, maltose, plant gel, 2,4 D, NAA, triacontanol, protein hydrolysate, taurine, biotin, plant sulfuration kinetin PSK α formed.Culture medium of the present invention has the advantages that to induce japonica rice anther callus expeditiously, can significantly improve japonica rice Anther Culture Efficiency.

Description

A kind of japonica rice flower pesticide Fiber differentiation based formulas

Technical field

The present invention relates to a kind of japonica rice flower pesticide Fiber differentiation based formulas, be specifically related to one and induce japonica rice flower pesticide expeditiously The Fiber differentiation based formulas of callus, belongs to technical field of agriculture science.

Background technology

Flower pesticide is the male organs of plant, and Anther Culture is to utilize plant tissue culture technique, growing to certain phase Flower pesticide, be inoculated on the culture medium of artificial preparation by aseptic technique, under a series of conditions are induced, change flower pesticide Development program, carries out mitosis, forms cell mass, and then forms callus, experience differentiation and development again after dedifferentiation Become the process of complete haplobiont.The pollen of plant is that pollen mother cell is formed through meiosis, its chromosome number The most somatic half, is called haploid cell.Make that pollen development therein becomes by the method for cultured in vitro flower pesticide one Whole plant, is called haplobiont, and therefore Breeding by anther culture is also haploid breeding.It is to train the most in vitro with flower pesticide Support, make pollen parthenogentic development become haplobiont, then become dliploid through chromosome doubling, therefrom select defect individual and be bred as New varieties.

1964, two botanist Guha and Maheshwari of Univ Delhi of India found, the hair leaf that they cultivate The embryoid grown in datura flower pesticide is derived from the haplobiont of pollen.Nineteen sixty-eight, Niizeki and Oono of Japan leads to Cross Anther Culture and obtain Haploid Rice plant.Along with deepening continuously of research, anther culture technique is progressively improved, and It is applied to Genetic and breeding in rice.Rice anther culture be more practical in current biotechnology breeding, breeding fast and effectively is new Technology.Rice Anther Culture Breeding main feature shows as shortening breeding cycle, improves efficiency of selection, accelerates effective character transfer Deng.Rice Anther Culture Breeding with conventional cross-breeding, distant hybridization breeding, mutation breeding and transgenic technology breeding phase In conjunction with, development defines a set of breeding technique system, plays Link role at traditional agriculture in the conversion of high-tech agricultural.

China's Research Work on Anther Culture in Rice starts from 1970, has obtained continuous improvement and complete in rice breeding is studied Kind, anther cultural efficiency is improved, and has cultivated a lot of indica rice kind and has passed through authorization and obtained spread.From Plants of Beijing research institute of the Chinese Academy of Sciences in 1975, crop breeding research institute of Heilongjiang Institute of Agricultural Sciences and Songhuajiang Area agriculture The haploid breeding cooperative groups of industry Science Institute paddy rice experiment centre composition is bred as japonica rice variety " single rich No. 1 " and 1976 first Since year Chinese Academy of Sciences's heredity institute and the Tianjin institute of agricultural sciences have been bred as " flower educates No. 1 " and " flower educates No. 2 ", China achieves and spends training Breeding to the breakthrough of application, also promotes the development of Anther culture breeding technology both domestic and external from theory.Conventional breeding and Anther Culture Technology combines and has been bred as a large amount of new rice variety by authorization, the relatively representational Chinese agriculture having the eighties in 20th century Institute of section " middle flower " series, wherein in spend No. 8, middle spend No. 9 and in spend No. 10 Annual planting areas once to reach 20,000 hm.The nineties " Jiangxi Xian " series of Jiangxi academy of agricultural sciences seed selection, Heilongjiang Institute of Agricultural Sciences's rice research has been selected imperial round-grained rice series of products, Different middle flower varieties and dragon round-grained rice kind is had again by successful seed selection after 2000.Through breeding research team in all parts of the country over more than 30 years 5 concentrate on studies, the existing large quantities of kind generations by Anther culture breeding of current China, and large scale application.Along with The researchs that deepen continuously such as anther cultural hereditary capacity, Physiology and biochemistry, present Rice anther culture has become as current biological skill New technology of breeding the most practical, effective in art breeding, the application study of China's Rice Anther Culture Breeding is the most always Hold the lead.

From the point of view of breed breeding, the regeneration plant that Anther Culture obtains is the most, it is thus achieved that the probability of superior genotypes is more Greatly.Growing of paddy rice is together decided on environmental factor by autogene, due to by nutrient media components, the difference of material The impact of property and culture environment etc., current Efficiency is the most preferable, it is thus achieved that regeneration plant colony little, anther culture descendant Phenotype is inadequate, and this makes selection, utilization be limited by bigger.Improve anther cultural induction frequency and Hua Pei green seedling frequency Rate, increases the colony of anther culture descendant, is the Anther Culture technology premise and the basis that expand application on genetic breeding.To this end, still So need to carry out unremitting effort.

Culture medium is anther cultural material base, is directly connected to growth and the differentiation of culture, and nutrient media components is Affect a critically important factor of Efficiency.The suitable amounts of each component of culture medium, and combination certain between them Relation, may result in increasing substantially of Efficiency.Continue screening and optimize minimal medium, improve culture medium select for Property, it is the effective measures improving Anther Culture Ability；Substantial amounts of research finds that some Organic additives can significantly improve flower pesticide training The power of supporting, finds and optimum organization Anther Culture Organic additives matter, can further improve Anther Culture Ability.

The process of Rice anther culture needs to carry out three steps: Fiber differentiation, will peel off at inducing culture by flower pesticide Upper first dedifferentiation induces anther callus；Differentiation is cultivated, and will transfer to divide on differential medium by anther callus again Metaplasia goes out to have the process of the green seedling of root and bud；Squamous subculture, the young plant that will differentiate is transferred on subculture medium Grow into the process of healthy and strong plant.Wherein, flower pesticide Fiber differentiation be the emphasis of Rice anther culture process be also difficult point, paddy rice The frequency of medicine callus induction and quality directly decide anther cultural success or failure.Therefore, combination and Optimal Medium component, Finding out efficient Rice Anther Fiber differentiation based formulas is to improve the key of Anther Culture induction frequency.

Groping by japonica rice for many years is anther cultural and puts into practice, we optimize minimal medium formula further, and Continuously attempt to add some organic additives improving japonica rice medicine induction force and combine the collocation of its kind and concentration, finally find out A kind of culture medium prescription inducing japonica rice flower pesticide Fiber differentiation expeditiously.Our result of study for further genralrlization and Application Rice Haploid Breeding and theoretical research are without being suspected to have bigger realistic meaning and practical value.

Summary of the invention

The invention provides a kind of culture medium for the japonica rice flower pesticide Fiber differentiation stage, the formula of this culture medium is as follows:

KNO₃900～1100mg/L, NH₄NO₃900～1100mg/L, Ca (NO₃)₂∙4H₂0 320～380mg/L, KH₂PO₄270～330mg/L, KCl 60～70mg/L, MgSO₄∙7H₂O 32～38mg/L, Na₂-EDTA 34～41mg/L, FeSO₄∙7H₂O 25～31mg/L, MnSO₄∙4H₂O 4.0～4.8mg/L, ZnSO₄∙7H₂O 1.35～1.65mg/L, H₃BO₃ 1.45～1.75mg/L, KI 0.7～0.9mg/L, inositol 90～110mg/L, glycerine 4.5～5.5mg/L, two water lemons Acid sodium 0.25～0.35mg/L, folic acid 0.25～0.35mg/L, glycine 1.8～2.2mg/L, lysine 1.8～ 2.2mg/L, vitamin B1 0.09～0.11mg/L, vitamin B6 0.09～0.11mg/L, nicotinic acid 0.45～0.55mg/L, Sucrose 38～42g/L, maltose 14～16g/L, plant gel 5～6g/L；

2,4-D 1.8～2.2mg/L, NAA 0.45～0.55mg/L, triacontanol 0.25～0.35mg/L, protein hydrolysate 0.27～0.33g/L, taurine 0.04～0.06mg/L, biotin 0.04～0.06mg/L, plant sulfuration kinetin PSK-α 15～25pmol/L, PH 5.8-6.0.

Culture medium of the present invention has the advantages that to induce japonica rice anther callus expeditiously, can significantly improve Japonica rice Anther Culture Efficiency.The following examples and contrast experiment can clearly reflect the spy of culture medium of the present invention Point.

Detailed description of the invention

Embodiment 1

It is formulated as follows the culture medium of formula:

KNO₃1000mg/L, NH₄NO₃1000mg/L, Ca (NO₃)₂∙4H₂0 350mg/L, KH₂PO₄300mg/L, KCl 65mg/L, MgSO₄∙7H₂O 35mg/L, Na₂-EDTA 38mg/L, FeSO₄∙7H₂O 28mg/L, MnSO₄∙4H₂O 4.4mg/L, ZnSO₄∙7H₂O 1.5mg/L, H₃BO₃1.6mg/L, KI 0.8mg/L, inositol 100mg/L, glycerine 5.0mg/L, two water lemon Lemon acid sodium 0.3mg/L, folic acid 0.3mg/L, glycine 2mg/L, lysine 2.0mg/L, vitamin B1 0.1mg/L, dimension is raw Element B6 0.1mg/L, nicotinic acid 0.5mg/L, sucrose 40g/L, maltose 15g/L, plant gel 5.5g/L；

2,4-D 2mg/L, NAA 0.5mg/L, triacontanol 0.3mg/L, protein hydrolysate 0.3g/L, taurine 0.05mg/L, biotin 0.05mg/L, plant sulfuration kinetin PSK-α 20pmol/L, PH 5.9.

Choose japonica rice variety and connect round-grained rice 3 and Xu rice No. 5 as Anther Culture flower pesticide donor, collection monokaryon flower pesticide in late period conduct Material to be tested, after surface sterilization, Cold pretreatment 3 days at 4 DEG C.After pretreatment, and outer implant sterilizing (0.1% mercuric chloride solution, 15 Min), trembling medicine method inoculation flower pesticide evoked callus in this inducing culture, each triangular flask 50 pieces of flower pesticide of inoculation, in temperature Be 28 DEG C, humidity be light culture in the environment of 60%-70%.(grow to about 2 mm) after Callus formation, calculate callus Inductivity.

Embodiment 2

It is formulated as follows the culture medium (optimal screening of carbon source combination mode) of formula:

KNO₃1000mg/L, NH₄NO₃1000mg/L, Ca (NO₃)₂∙4H₂0 350mg/L, KH₂PO₄300mg/L, KCl 65mg/L, MgSO₄∙7H₂O 35mg/L, Na₂-EDTA 38mg/L, FeSO₄∙7H₂O 28mg/L, MnSO₄∙4H₂O 4.4mg/L, ZnSO₄∙7H₂O 1.5mg/L, H₃BO₃1.6mg/L, KI 0.8mg/L, inositol 100mg/L, glycerine 5.0mg/L, two water lemon Lemon acid sodium 0.3mg/L, folic acid 0.3mg/L, glycine 2mg/L, lysine 2.0mg/L, vitamin B1 0.1mg/L, dimension is raw Element B6 0.1mg/L, nicotinic acid 0.5mg/L, plant gel 5.5g/L；

2,4-D 2mg/L, NAA 0.5mg/L, triacontanol 0.3mg/L, protein hydrolysate 0.3g/L, taurine 0.05mg/L, biotin 0.05mg/L, plant sulfuration kinetin PSK-α 20pmol/L, PH 5.9.

Carbon source combination mode arranges following 11 kinds: maltose 55g/L；Sucrose 5g/L, maltose 50g/L；Sucrose 10g/L, maltose 45g/L；Sucrose 15g/L, maltose 40g/L；Sucrose 20g/L, maltose 35g/L；Sucrose 25g/ L, maltose 30g/L；Sucrose 30g/L, maltose 25g/L；Sucrose 35g/L, maltose 20g/L；Sucrose 45g/L, wheat Bud sugar 10g/L；Sucrose 50g/L, maltose 5g/L；Sucrose 55g/L.

Choose these two japonica rice varieties equally to heal with embodiment 1, calculating as flower pesticide donor, method of operating, cultural method Injured tissue inductivity.

Embodiment 3

It is formulated as follows the culture medium (optimal screening of coagulator) of formula:

KNO₃1000mg/L, NH₄NO₃1000mg/L, Ca (NO₃)₂∙4H₂0 350mg/L, KH₂PO₄300mg/L, KCl 65mg/L, MgSO₄∙7H₂O 35mg/L, Na₂-EDTA 38mg/L, FeSO₄∙7H₂O 28mg/L, MnSO₄∙4H₂O 4.4mg/L, ZnSO₄∙7H₂O 1.5mg/L, H₃BO₃1.6mg/L, KI 0.8mg/L, inositol 100mg/L, glycerine 5.0mg/L, two water lemon Lemon acid sodium 0.3mg/L, folic acid 0.3mg/L, glycine 2mg/L, lysine 2.0mg/L, vitamin B1 0.1mg/L, dimension is raw Element B6 0.1mg/L, nicotinic acid 0.5mg/L, sucrose 40g/L, maltose 15g/L；

2,4-D 2mg/L, NAA 0.5mg/L, triacontanol 0.3mg/L, protein hydrolysate 0.3g/L, taurine 0.05mg/L, biotin 0.05mg/L, plant sulfuration kinetin PSK-α 20pmol/L, PH 5.9.

Coagulator is set to following 2 kinds: plant gel 2.75g/L, agar 3.5g；Agar 7g.

Choose these two japonica rice varieties equally to heal with embodiment 1, calculating as flower pesticide donor, method of operating, cultural method Injured tissue inductivity.

Embodiment 4

It is formulated as follows the culture medium (hormone combinations and the optimal screening of matched proportion density) of formula:

KNO₃1000mg/L, NH₄NO₃1000mg/L, Ca (NO₃)₂∙4H₂0 350mg/L, KH₂PO₄300mg/L, KCl 65mg/L, MgSO₄∙7H₂O 35mg/L, Na₂-EDTA 38mg/L, FeSO₄∙7H₂O 28mg/L, MnSO₄∙4H₂O 4.4mg/L, ZnSO₄∙7H₂O 1.5mg/L, H₃BO₃1.6mg/L, KI 0.8mg/L, inositol 100mg/L, glycerine 5.0mg/L, two water lemon Lemon acid sodium 0.3mg/L, folic acid 0.3mg/L, glycine 2mg/L, lysine 2.0mg/L, vitamin B1 0.1mg/L, dimension is raw Element B6 0.1mg/L, nicotinic acid 0.5mg/L, sucrose 40g/L, maltose 15g/L, plant gel 5.5g/L；

Protein hydrolysate 0.3g/L, taurine 0.05mg/L, biotin 0.05mg/L, plant sulfuration kinetin PSK-α 20pmol/L, PH 5.9.

Hormone combinations and matched proportion density are set to two categories below:

One class fixes triacontanol 0.3mg/L, 2,4D, NAA combination following 6 kinds: 2,4D 0.5mg/L, NAA are set 0.5mg/L；2,4D 1mg/L, NAA 0.5mg/L；2,4D 3mg/L, NAA 0.5mg/L；2,4D 2mg/L, NAA 0.25mg/ L；2,4D 2mg/L, NAA 0.75mg/L；2,4D 2mg/L, NAA 1mg/L；

Two classes fix 2,4-D 2mg/L, NAA 0.5mg/L, and triacontanol adds concentration and arranges following 6 kinds: 0；0.1g/L； 0.2g/L；0.4g/L；0.5g/L；0.8g/L.

Choose these two japonica rice varieties equally to heal with embodiment 1, calculating as flower pesticide donor, method of operating, cultural method Injured tissue inductivity.

Embodiment 5

It is formulated as follows the culture medium optimal screening of concentration (protein hydrolysate add) of formula:

KNO₃1000mg/L, NH₄NO₃1000mg/L, Ca (NO₃)₂∙4H₂0 350mg/L, KH₂PO₄300mg/L, KCl 65mg/L, MgSO₄∙7H₂O 35mg/L, Na₂-EDTA 38mg/L, FeSO₄∙7H₂O 28mg/L, MnSO₄∙4H₂O 4.4mg/L, ZnSO₄∙7H₂O 1.5mg/L, H₃BO₃1.6mg/L, KI 0.8mg/L, inositol 100mg/L, glycerine 5.0mg/L, two water lemon Lemon acid sodium 0.3mg/L, folic acid 0.3mg/L, glycine 2mg/L, lysine 2.0mg/L, vitamin B1 0.1mg/L, dimension is raw Element B6 0.1mg/L, nicotinic acid 0.5mg/L, sucrose 40g/L, maltose 15g/L, plant gel 5.5g/L；

2,4-D 2mg/L, NAA 0.5mg/L, triacontanol 0.3mg/L, taurine 0.05mg/L, biotin 0.05mg/L, plant sulfuration kinetin PSK-α 20pmol/L, PH 5.9.

Protein hydrolysate adds concentration and arranges following 6 kinds: 0；0.1 g/L；0.2g/L；0. 4g/L；0.5 g/L；0.8 g/L.

Choose these two japonica rice varieties equally to heal with embodiment 1, calculating as flower pesticide donor, method of operating, cultural method Injured tissue inductivity.

Embodiment 6

It is formulated as follows the culture medium optimal screening of kinetin PSK α (plant sulfuration) of formula:

KNO₃1000mg/L, NH₄NO₃1000mg/L, Ca (NO₃)₂∙4H₂0 350mg/L, KH₂PO₄300mg/L, KCl 65mg/L, MgSO₄∙7H₂O 35mg/L, Na₂-EDTA 38mg/L, FeSO₄∙7H₂O 28mg/L, MnSO₄∙4H₂O 4.4mg/L, ZnSO₄∙7H₂O 1.5mg/L, H₃BO₃1.6mg/L, KI 0.8mg/L, inositol 100mg/L, glycerine 5.0mg/L, two water lemon Lemon acid sodium 0.3mg/L, folic acid 0.3mg/L, glycine 2mg/L, lysine 2.0mg/L, vitamin B1 0.1mg/L, dimension is raw Element B6 0.1mg/L, nicotinic acid 0.5mg/L, sucrose 40g/L, maltose 15g/L, plant gel 5.5g/L；

2,4-D 2mg/L, NAA 0.5mg/L, triacontanol 0.3mg/L, protein hydrolysate 0.3g/L, taurine 0.05mg/L, biotin 0.05mg/L, PH 5.9.

Plant sulfuration kinetin PSK α adds concentration and arranges following 6 kinds: 0；10pmol/L；30pmol/L；40pmol/L； 60pmol/L；80pmol/L.

Choose these two japonica rice varieties equally to heal with embodiment 1, calculating as flower pesticide donor, method of operating, cultural method Injured tissue inductivity.

Culture medium contrast experiment

Embodiment 2-6 being contrasted with embodiment 1 respectively, the result of contrast is as shown in the table:

From table 1-6 it can be seen that use culture medium provided by the present invention than using the induction of other any contrast culture medium Japonica rice anther callus in hgher efficiency, shows that the culture medium prescription that the present invention provides is through a large amount of single-factors, multiple-factor The optimum combination that test is screened, we have also made the change optimization of little scope single-factor around the combination matching of the present invention for many years Composite test, substantial amounts of experimental study also show that the component proportion of the present invention is optimum.Use cultivation provided by the present invention Base connects round-grained rice 3, the slowly callus induction rate of rice No. 5 and is up to 28.7% and 26.9% japonica rice variety, has absolutely proved the present invention The culture medium provided is a kind of excellent japonica rice flower pesticide inducing culture.

Embodiment 7

In order to the culture medium that relatively culture medium provided by the present invention and forefathers use induces japonica rice antherderived callus effect Difference, we have searched 6 japonica rice antherderived callus Fiber differentiation based formulas from pertinent literature, have been configured to antherderived callus Fiber differentiation Base, chooses japonica rice variety equally and connects round-grained rice 3, slowly rice No. 5 as flower pesticide donor induction antherderived callus, method of operating, cultural method With embodiment 1, calculate callus induction rate.

Other 6 inducing cultures are:

MS minimal medium+3% sucrose, 3% maltose+2,4-D(2mg/L), NAA(2mg/L)+agar (7g/L), pH " screenings of high Anther Culture Efficiency japonica rice cross combination " such as Wang Qing is come from) for 5.8(documents；

N6 minimal medium+3% sucrose, 3% maltose+2,4-D(2mg/L), NAA(2mg/L)+agar (7g/L), pH " screenings of high Anther Culture Efficiency japonica rice cross combination " such as Wang Qing is come from) for 5.8(documents；

Improvement N6 culture medium prescription: KNO₃2830mg/L, (NH₄)₂SO₄463mg/L, KH₂PO₄400mg/L, MgSO₄∙ 7H₂O 185mg/L, CaCl₂∙2H₂O 166mg/L, MnSO₄∙4H₂O 4.4mg/L, ZnSO₄∙7H₂O 1.55mg/L, H₃BO₃ 1.6mg/L, KI 0.8mg/L, CuSO₄∙5H₂O 0.025mg/L, CoCl₂∙6H₂O 0.025mg/L, Na₂MoO₄∙2H₂O 0.25mg/L, Fe-EDTA 5.57mg/L, glycine 2mg/L, vitamin B1 1mg/L, vitamin B6 0.5mg/L, nicotinic acid 0.5mg/L, sucrose 50g/L, glucitol 100 mg/L, LH 500mg/L, 2,4-D 2mg/L, agar 6g, pH are 5.8(pair " researchs of japonica rice anther culture medium Optimum Experiment " such as the honor coming from grandson than file)；

SK3 minimal medium+2,4-D(2mg/L), NAA(1mg/L), KT(1mg/L), LH(0.5 g/L)+sucrose (50 G/L)+agar (4.2g/L), pH is that 5.8(documents comes from seedling " North Japonica Rice different genotype Anther Culture spy such as vertical new grade Property compares ")；

M8 minimal medium+3% sucrose, 3% maltose+2,4-D(2mg/L), NAA(2mg/L)+agar (7g/L), pH " screenings of high Anther Culture Efficiency japonica rice cross combination " such as Wang Qing is come from) for 5.8(documents；

B5 minimal medium+3% sucrose, 3% maltose+2,4-D(2mg/L), NAA(2mg/L)+agar (7g/L), pH " screenings of high Anther Culture Efficiency japonica rice cross combination " such as Wang Qing is come from) for 5.8(documents.

Culture medium contrast experiment

Embodiment 7 being contrasted with embodiment 1, the result of contrast is as shown in the table:

From the point of view of the inducing effect of different culture media, culture medium of the present invention inductivity average to japonica rice anther callus is 27.8%, MS, N6, improvement N6, SK3, M8, B5 medium are then 2.4%, 14.8%, 20.0%, 15.6%, 8.8%, 3.8%, Ke Yifa The existing present invention to be significantly higher than other 6 kinds of inducing cultures to the inductivity of japonica rice anther callus.

Above 7 embodiments can illustrate that the component proportion of the culture medium prescription that the present invention provides is optimum combination, this The japonica rice flower pesticide inducing culture that the induction ratio forefathers of japonica rice anther callus are found by the culture medium of bright offer has substantially Advantage.Our result of study is for further genralrlization and applies haploid breeding without being suspected to have bigger realistic meaning and practical valency Value.

Those skilled in the art can be according to present disclosure and the art technology grasped in the present invention Holding and make replacement or modification, but these are replaced or modification is all not regarded as a departure from present inventive concept, these are replaced or modification All in claimed interest field.

Claims (1)

1., for the culture medium in japonica rice flower pesticide Fiber differentiation stage, the formula of this culture medium is as follows:

KNO₃900～1100mg/L, NH₄NO₃900～1100mg/L, Ca (NO₃)₂∙4H₂0 320～380mg/L, KH₂PO₄ 270～330mg/L, KCl 60～70mg/L, MgSO₄∙7H₂O 32～38mg/L, Na₂-EDTA 34～41mg/L, FeSO₄∙ 7H₂O 25～31mg/L, MnSO₄∙4H₂O 4.0～4.8mg/L, ZnSO₄∙7H₂O 1.35～1.65mg/L, H₃BO₃1.45～ 1.75mg/L, KI 0.7～0.9mg/L, inositol 90～110mg/L, glycerine 4.5～5.5mg/L, Sodium Citrate, usp, Dihydrate Powder 0.25～0.35mg/L, folic acid 0.25～0.35mg/L, glycine 1.8～2.2mg/L, lysine 1.8～2.2mg/L, dimension Raw element B1 0.09～0.11mg/L, vitamin B6 0.09～0.11mg/L, nicotinic acid 0.45～0.55mg/L, sucrose 38～ 42g/L, maltose 14～16g/L, plant gel 5～6g/L；

2,4-D 1.8～2.2mg/L, NAA 0.45～0.55mg/L, triacontanol 0.25～0.35mg/L, protein hydrolysate 0.27～0.33g/L, taurine 0.04～0.06mg/L, biotin 0.04～0.06mg/L, plant sulfuration kinetin PSK-α 15～25pmol/L, PH 5.8-6.0.