CN104705193A - Formula of induction culture medium for japonica rice anthers - Google Patents

Abstract

The invention provides a culture medium for induction culture stage for japonica rice anthers. A formula of the culture medium consists of the following ingredients in a certain ratio: KNO3, NH4NO3, Ca(NO3)2.4H20, KH2PO4, KCl, MgSO4.7H2O, Na2-EDTA, FeSO4.7H2O, MnSO4.4H2O, ZnSO4.7H2O, H3BO3, KI, inosite, glycerol, Na3C6H5O7.H2O, folic acid, glycine, lysine, vitamin B1, vitamin B6, nicotinic acid, saccharose, maltose, plant gel, 2,4-D, NAA, triacontanol, hydrolyzed protein, taurine, biotin and plant vulcanization kinetin PSK-alpha. The culture medium provided by the invention has the characteristic of efficiently inducing japonica rice anther caluus and can be used for remarkably increasing the culture efficiency of the japonica rice anthers.

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 a kind of Fiber differentiation based formulas of inducing japonica rice anther callus expeditiously, belong to technical field of agriculture science.

Background technology

Flower pesticide is the male organs of plant, anther culture utilizes plant tissue culture technique, the flower pesticide of growing to certain phase, be inoculated on the medium of artificial preparation by aseptic technique, under a series of condition induction, change the development program of flower pesticide, carry out mitosis, after dedifferentiation, form cell mass, and then formation callus, experience the process that differentiation and development again becomes complete haplobiont.The pollen of plant is that pollen mother cell is formed through reduction division, and its chromosome number only has somatic half, is called haploid cell.With the whole plant that the method for cultured in vitro flower pesticide makes pollen development wherein become, be called haplobiont, therefore Breeding by anther culture is also haploid breeding.It is by the artificial cultured in vitro of flower pesticide, makes pollen parthenogentic development become haplobiont, then becomes dliploid through chromosome doubling, therefrom selects defect individual and is bred as new varieties.

1964, two botanist Guha and Maheshwari of Univ Delhi of India found, the embryoid grown in the hair leaf datura flower pesticide that they cultivate is the haplobiont deriving from pollen.Nineteen sixty-eight, Niizeki and Oono of Japan obtains Haploid Rice plant by anther culture.Along with deepening continuously of research, anther culture technique is progressively improved, and is applied to Genetic and breeding in rice.Rice anther culture is comparatively practical in current biotechnology breeding, new technology of breeding fast and effectively.Rice Anther Culture Breeding main feature shows as shortening breeding cycle, improves efficiency of selection, accelerates effective character transfer etc.Rice Anther Culture Breeding combines with conventional cross-breeding, distant hybridization breeding, mutation breeding and transgenic technology breeding, and development defines a set of breeding technique system, in the conversion of traditional farming to high-tech agricultural, play Link role.

China's Research Work on Anther Culture in Rice starts from 1970, and in rice breeding research, obtain constantly improvement and perfect, anther cultural efficiency is improved, and has cultivated a lot of indica rice kind and have passed and authorize and obtain spread.Be bred as since japonica rice variety " single rich No. 1 " and heredity institutes of the Chinese Academy of Sciences in 1976 and the Tianjin institute of agricultural sciences be bred as " flower educates No. 1 " and " spend and educate No. 2 " first from the haploid breeding cooperative groups of 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 Institute of agricultural sciences paddy rice experiment centre composition, China achieves Anther culture breeding from theory to the breakthrough of application, also promotes the development of Anther culture breeding technology both domestic and external.Conventional breeding combines with anther culture technique and has been bred as a large amount of new rice variety by authorizing, more representational " middle flower " series having a Chinese Academy of Agricultural Sciences of the eighties in 20th century, spend wherein No. 8, middle spend No. 9 and in spend No. 10 Annual planting areas once to reach 20,000 hm." Jiangxi Xian " series of the Jiangxi academy of agricultural sciences seed selection of the nineties, Heilongjiang Institute of Agricultural Sciences's rice research has selected imperial round-grained rice series of products, has again different middle flower varieties and imperial round-grained rice kind by successful seed selection after 2000.Through concentrating on studies of breeding research troop in all parts of the country over more than 30 years, current China has large quantities of kind by Anther culture breeding and produces, and large scale application.Along with researchs that deepens continuously such as anther cultural hereditary capacity, Physiology and biochemistries, present Rice anther culture has become comparatively practical, effective new technology of breeding in current biotechnology breeding, and the application study of China's Rice Anther Culture Breeding is held the lead in the world always.

From the angle of breed breeding, the regeneration plant that anther culture obtains is more, and the probability obtaining superior genotypes is larger.Growing of paddy rice is determined jointly by autogene and environmental factor, due to the impact by nutrient media components, the otherness of material and culture environment etc., current Efficiency is desirable not enough, the regeneration plant colony obtained is little, the phenotype of anther culture descendant is inadequate, and this makes selection, utilization is subject to larger restriction.Improve anther cultural induction frequency and the green seedling frequency of Hua Pei, increasing the colony of anther culture descendant, is prerequisite and the basis that Anther Culture technology expands application on genetic breeding.For this reason, still need to carry out unremitting effort.

Medium is anther cultural material base, is directly connected to the Proliferation and differentiation of culture, and nutrient media components affects the very important factor of of Efficiency.The suitable amounts of each component of medium, and syntagmatic certain between them, can cause increasing substantially of Efficiency.Continuing screening and optimize minimal medium, improving the specific aim that medium is selected, is the effective measures improving Anther Culture Ability; Large quantifier elimination finds that some Organic additives can significantly improve Anther Culture Ability, finds and optimum organization anther culture Organic additives matter, can further improve Anther Culture Ability.

The process need of Rice anther culture carries out three steps: Fiber differentiation, peels off first dedifferentiation on inducing culture induce anther callus by flower pesticide; Differentiation is cultivated, and transfers to the process of differential medium breaking up again and bearing the green seedling with root and bud by anther callus; Squamous subculture, the young plant being about to differentiate is transferred to process subculture medium growing into healthy and strong plant.Wherein, flower pesticide Fiber differentiation is the emphasis of Rice anther culture process is also difficult point, and the frequency of Rice Anther 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 improve the key of anther culture induction frequency.

Grope by japonica rice is anther cultural for many years and puts into practice, we optimize minimal medium formula further, and constantly attempt adding some organic additives improving japonica rice medicine induction force and combining the collocation of its kind and concentration, finally found out a kind of culture medium prescription of inducing japonica rice flower pesticide Fiber differentiation expeditiously.Our result of study for further genralrlization and application Rice Haploid Breeding and theoretical research without being suspected to have larger realistic meaning and practical value.

Summary of the invention

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

KNO ₃900 ~ 1100mg/L, NH ₄nO ₃900 ~ 1100mg/L, Ca (NO ₃) ₂4H ₂0 320 ~ 380mg/L, KH ₂pO ₄270 ~ 330mg/L, KCl 60 ~ 70mg/L, MgSO ₄7H ₂o 32 ~ 38mg/L, Na ₂-EDTA 34 ~ 41mg/L, FeSO ₄7H ₂o 25 ~ 31mg/L, MnSO ₄4H ₂o 4.0 ~ 4.8mg/L, ZnSO ₄7H ₂o 1.35 ~ 1.65mg/L, H ₃bO ₃1.45 ~ 1.75mg/L, KI 0.7 ~ 0.9mg/L, inositol 90 ~ 110mg/L, glycerine 4.5 ~ 5.5mg/L, Na ₃c ₆h ₅o ₇h ₂o 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; vitamin h 0.04 ~ 0.06mg/L, plant sulfuration kinetin PSK-α 15 ~ 25pmol/L, PH 5.8-6.0.

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 clearly can reflect the feature of medium of the present invention.

Embodiment

Embodiment 1

Be formulated as follows the medium of formula:

KNO ₃1000mg/L, NH ₄nO ₃1000mg/L, Ca (NO ₃) ₂4H ₂0 350mg/L, KH ₂pO ₄300mg/L, KCl 65mg/L, MgSO ₄7H ₂o 35mg/L, Na ₂-EDTA 38mg/L, FeSO ₄7H ₂o 28mg/L, MnSO ₄4H ₂o 4.4mg/L, ZnSO ₄7H ₂o 1.5mg/L, H ₃bO ₃1.6mg/L, KI 0.8mg/L, inositol 100mg/L, glycerine 5.0mg/L, Na ₃c ₆h ₅o ₇h ₂o 0.3mg/L, folic acid 0.3mg/L, glycine 2mg/L, lysine 2.0mg/L, vitamin B1 0.1mg/L, vitamin 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, vitamin h 0.05mg/L, plant sulfuration kinetin PSK-α 20pmol/L, PH 5.9.

Choose japonica rice variety and connect No. 3, round-grained rice with Xu rice No. 5 as anther culture flower pesticide donor, collection monokaryon flower pesticide in late period tries material as supplying, after surface sterilization, and Cold pretreatment 3 days at 4 DEG C.After pretreatment, explant sterilization (0.1% mercuric chloride solution, 15 min), tremble medicine method inoculation flower pesticide evoked callus in this inducing culture, each triangular flask inoculates 50 pieces of flower pesticide, temperature be 28 DEG C, humidity be the environment of 60%-70% under light culture.(grow to about 2 mm) after Callus formation, calculate callus induction rate.

Embodiment 2

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

KNO ₃1000mg/L, NH ₄nO ₃1000mg/L, Ca (NO ₃) ₂4H ₂0 350mg/L, KH ₂pO ₄300mg/L, KCl 65mg/L, MgSO ₄7H ₂o 35mg/L, Na ₂-EDTA 38mg/L, FeSO ₄7H ₂o 28mg/L, MnSO ₄4H ₂o 4.4mg/L, ZnSO ₄7H ₂o 1.5mg/L, H ₃bO ₃1.6mg/L, KI 0.8mg/L, inositol 100mg/L, glycerine 5.0mg/L, Na ₃c ₆h ₅o ₇h ₂o 0.3mg/L, folic acid 0.3mg/L, glycine 2mg/L, lysine 2.0mg/L, vitamin B1 0.1mg/L, vitamin 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, vitamin h 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, maltose 10g/L; Sucrose 50g/L, maltose 5g/L; Sucrose 55g/L.

Choose these two japonica rice varieties equally as flower pesticide donor, method of operating, cultural method, with embodiment 1, calculate callus induction rate.

Embodiment 3

Be formulated as follows the medium (optimal screening of coagulating agent) of formula:

KNO ₃1000mg/L, NH ₄nO ₃1000mg/L, Ca (NO ₃) ₂4H ₂0 350mg/L, KH ₂pO ₄300mg/L, KCl 65mg/L, MgSO ₄7H ₂o 35mg/L, Na ₂-EDTA 38mg/L, FeSO ₄7H ₂o 28mg/L, MnSO ₄4H ₂o 4.4mg/L, ZnSO ₄7H ₂o 1.5mg/L, H ₃bO ₃1.6mg/L, KI 0.8mg/L, inositol 100mg/L, glycerine 5.0mg/L, Na ₃c ₆h ₅o ₇h ₂o 0.3mg/L, folic acid 0.3mg/L, glycine 2mg/L, lysine 2.0mg/L, vitamin B1 0.1mg/L, vitamin 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, vitamin h 0.05mg/L, plant sulfuration kinetin PSK-α 20pmol/L, PH 5.9.

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

Choose these two japonica rice varieties equally as flower pesticide donor, method of operating, cultural method, with embodiment 1, calculate callus induction rate.

Embodiment 4

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

KNO ₃1000mg/L, NH ₄nO ₃1000mg/L, Ca (NO ₃) ₂4H ₂0 350mg/L, KH ₂pO ₄300mg/L, KCl 65mg/L, MgSO ₄7H ₂o 35mg/L, Na ₂-EDTA 38mg/L, FeSO ₄7H ₂o 28mg/L, MnSO ₄4H ₂o 4.4mg/L, ZnSO ₄7H ₂o 1.5mg/L, H ₃bO ₃1.6mg/L, KI 0.8mg/L, inositol 100mg/L, glycerine 5.0mg/L, Na ₃c ₆h ₅o ₇h ₂o 0.3mg/L, folic acid 0.3mg/L, glycine 2mg/L, lysine 2.0mg/L, vitamin B1 0.1mg/L, vitamin 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, vitamin h 0.05mg/L, plant sulfuration kinetin PSK-α 20pmol/L, PH 5.9.

Hormone combinations and matched proportion density are set to following two classes:

One class fixes triacontanol 0.3mg/L, and 2,4D, NAA combination arrange following 6 kinds: 2,4D 0.5mg/L, NAA 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 as flower pesticide donor, method of operating, cultural method, with embodiment 1, calculate callus induction rate.

Embodiment 5

Be formulated as follows the medium (protein hydrolysate adds the optimal screening of concentration) of formula:

KNO ₃1000mg/L, NH ₄nO ₃1000mg/L, Ca (NO ₃) ₂4H ₂0 350mg/L, KH ₂pO ₄300mg/L, KCl 65mg/L, MgSO ₄7H ₂o 35mg/L, Na ₂-EDTA 38mg/L, FeSO ₄7H ₂o 28mg/L, MnSO ₄4H ₂o 4.4mg/L, ZnSO ₄7H ₂o 1.5mg/L, H ₃bO ₃1.6mg/L, KI 0.8mg/L, inositol 100mg/L, glycerine 5.0mg/L, Na ₃c ₆h ₅o ₇h ₂o 0.3mg/L, folic acid 0.3mg/L, glycine 2mg/L, lysine 2.0mg/L, vitamin B1 0.1mg/L, vitamin 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, vitamin h 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 as flower pesticide donor, method of operating, cultural method, with embodiment 1, calculate callus induction rate.

Embodiment 6

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

KNO ₃1000mg/L, NH ₄nO ₃1000mg/L, Ca (NO ₃) ₂4H ₂0 350mg/L, KH ₂pO ₄300mg/L, KCl 65mg/L, MgSO ₄7H ₂o 35mg/L, Na ₂-EDTA 38mg/L, FeSO ₄7H ₂o 28mg/L, MnSO ₄4H ₂o 4.4mg/L, ZnSO ₄7H ₂o 1.5mg/L, H ₃bO ₃1.6mg/L, KI 0.8mg/L, inositol 100mg/L, glycerine 5.0mg/L, Na ₃c ₆h ₅o ₇h ₂o 0.3mg/L, folic acid 0.3mg/L, glycine 2mg/L, lysine 2.0mg/L, vitamin B1 0.1mg/L, vitamin 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, vitamin h 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 as flower pesticide donor, method of operating, cultural method, with embodiment 1, calculate callus induction rate.

Medium contrast experiment

Contrasted with embodiment 1 respectively by embodiment 2-6, the result of contrast is as shown in the table:

As can be seen from table 1-6, it is higher than using the efficiency of other any contrast medium induction japonica rice anther callus to use medium provided by the present invention, show the optimum combination that culture medium prescription provided by the invention is through a large amount of single-factor, multifactor experiment screens, we have also made single-factor change optimum organization test among a small circle around combination matching of the present invention for many years, and a large amount of experimental studies also show that component proportion of the present invention is optimum.Use medium provided by the present invention to connect the callus induction rate of No. 3, round-grained rice, slowly rice No. 5 up to 28.7% and 26.9% to japonica rice variety, absolutely proved that medium provided by the invention is a kind of excellent japonica rice flower pesticide inducing culture.

Embodiment 7

The medium adopted in order to medium more provided by the present invention and forefathers induces the difference of japonica rice antherderived callus effect, we have searched 6 japonica rice antherderived callus Fiber differentiation based formulas from pertinent literature, be mixed with antherderived callus inducing culture, choose japonica rice variety equally and connect No. 3, round-grained rice, Xu 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 is that 5.8(documents comes from " screenings of high Anther Culture Efficiency japonica rice hybrid combination " such as Wang Qing);

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

Improvement N6 culture medium prescription: KNO ₃2830mg/L, (NH ₄) ₂sO ₄463mg/L, KH ₂pO ₄400mg/L, MgSO ₄7H ₂o 185mg/L, CaCl ₂2H ₂o 166mg/L, MnSO ₄4H ₂o 4.4mg/L, ZnSO ₄7H ₂o 1.55mg/L, H ₃bO ₃1.6mg/L, KI 0.8mg/L, CuSO ₄5H ₂o 0.025mg/L, CoCl ₂6H ₂o 0.025mg/L, Na ₂moO ₄2H ₂o 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 that 5.8(documents comes from " researchs of japonica rice anther culture medium Optimum Experiment " such as the honor of grandson);

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 that seedling is vertical new waits " North Japonica Rice different genotype Anther culture characteristics compares ");

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

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

Medium contrast experiment

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

From the inducing effect of different culture media, medium of the present invention is 27.8% to the average inductivity of japonica rice anther callus, MS, N6, improvement N6, SK3, M8, B5 medium are then 2.4%, 14.8%, 20.0%, 15.6%, 8.8%, 3.8%, can find that the present invention will 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 culture medium prescription provided by the invention is optimum combination, and medium provided by the invention has clear superiority to the japonica rice flower pesticide inducing culture that the induction ratio forefathers of japonica rice anther callus find.Our result of study for further genralrlization and application haploid breeding without being suspected to have larger realistic meaning and practical value.

Those skilled in the art can make replacement or modification according to content disclosed by the invention and the art technology grasped to content of the present invention; but these replacements or modification should not be considered as disengaging the present invention design, and these replacements or modification are all in the interest field of application claims protection.

Claims (1)

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

KNO ₃900 ~ 1100mg/L, NH ₄nO ₃900 ~ 1100mg/L, Ca (NO ₃) ₂4H ₂0 320 ~ 380mg/L, KH ₂pO ₄270 ~ 330mg/L, KCl 60 ~ 70mg/L, MgSO ₄7H ₂o 32 ~ 38mg/L, Na ₂-EDTA 34 ~ 41mg/L, FeSO ₄7H ₂o 25 ~ 31mg/L, MnSO ₄4H ₂o 4.0 ~ 4.8mg/L, ZnSO ₄7H ₂o 1.35 ~ 1.65mg/L, H ₃bO ₃1.45 ~ 1.75mg/L, KI 0.7 ~ 0.9mg/L, inositol 90 ~ 110mg/L, glycerine 4.5 ~ 5.5mg/L, Na ₃c ₆h ₅o ₇h ₂o 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; vitamin h 0.04 ~ 0.06mg/L, plant sulfuration kinetin PSK-α 15 ~ 25pmol/L, PH 5.8-6.0.