CN110810244A - Culture medium for tissue culture of floral rod - Google Patents
Culture medium for tissue culture of floral rod Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/22—Improving land use; Improving water use or availability; Controlling erosion
Abstract
The invention provides a culture medium for tissue culture of a floral rod, which comprises a primary generation induction culture medium, a secondary generation culture medium and a rooting culture medium, and can be used for taking a growing point on a tender branch of the floral rod as an explant, and performing the processes of primary generation induction culture, secondary generation culture, rooting culture and transplanting after sterilization. The optimal culture medium formula and the combination thereof selected in each growth period in the whole tissue culture process of the flower stick can fully meet the nutritional requirements and growth development of the flower stick in each period, promote the whole tissue culture propagation process of the flower stick, ensure the successful implementation of the tissue culture of the flower stick and provide powerful support for the large-scale production of the flower stick. The culture medium has the advantages of simple formula, low cost and good application prospect.
Description
Technical Field
The invention belongs to the technical field of plant biology, and particularly relates to a culture medium for each stage in tissue culture of a floral rod. The optimal culture medium for different periods in tissue culture of the flower stick is screened out, the proportion of each hormone in the culture medium is determined, and the culture medium prepared according to the formula can meet the nutritional requirements and growth development of the flower stick at each period.
Background
The floral rod is also called as the root of membranous milkvetch, flower firewood, flower stem, flower bud, floral cap and oxtail tip, and is a typical sand-fixing pioneer plant in the desert region of the large deciduous shrub of the genus Astragalus of the subfamily Papilionaceae of Leguminosae. The artificial planting method is characterized in that the artificial planting method is distributed in arid and semi-arid desert or sand land in middle Asia, and natural groups exist in Guerbantong Gute desert, Ba Dan Jilin desert, Hexi corridor sand land, Tenggeli desert, Wulan cloth and desert in China and west desert in Kubu, and artificial planting groups and descendants thereof exist in other deserts or sand lands. The eastern sandy land is used for introducing the flower sticks, and the growth is good and even superior to that of a natural distribution area. The flower stick is an excellent sand-fixing shrub, has certain economic benefit, and has large nursery stock demand in the ecological civilization construction process of a sand area.
Plant tissue culture is an important biotechnological means. Cells, stem segments, leaves, etc. of the plant may be cultured. The cells, organs and tissues of the plant are taken and put into a container with proper culture medium, and can be differentiated to form a complete plant under certain culture conditions. Tissue culture as a powerful means of biotechnology is increasingly paid attention to, and plays an important role in aspects of detoxification, rapid propagation, mutation induction, cell engineering, genetic engineering and the like of agricultural and forestry crops.
Under the condition of in vitro culture, tissue cells of different plants and different parts of the same plant have different nutritional requirements, and can grow and develop better only if the special requirements of the different plants and the tissue cells of different parts of the same plant are met. Mastering the preparation and screening method of the culture medium is one of the key links for obtaining the success of tissue culture.
The method is an effective way for solving ecological seedling use of the flower sticks by improving the quantity of industrial flower stick seedlings through a tissue culture propagation technology. However, in the prior art, tissue culture of the floral rod is still in a groping stage, few reference materials can be used in the preparation of the technical scheme, the reference materials are required to be solved one by one through experiments, and the key points are to solve the problems of explant material taking part and period, culture medium selection in each culture period, culture conditions, rapid propagation technology after seedling formation and the like. The invention aims at developing and utilizing, reducing cost and forming industrial and industrialized production, and makes corresponding working plan and implementation method
The whole process of tissue culture is generally divided into several technical links such as establishment of a culture scheme, selection and treatment of explants, inoculation, primary culture, subculture, strong seedling and rooting culture, domestication and transplantation of test-tube seedlings and the like. Under the condition of in vitro culture, tissue cells of different plants and different parts of the same plant have different nutritional requirements, and can grow and develop better only if the special requirements of the different plants and the tissue cells of different parts of the same plant are met. The control of the preparation and screening method of the culture medium is one of the key links for successful tissue culture.
Disclosure of Invention
The invention aims to screen out the optimal culture medium for different periods in tissue culture of a flower stick, determine the proportion of each hormone in the culture medium, and the culture medium prepared according to the formula can meet the nutritional requirements and growth development of the flower stick at each period.
The invention sets an implementation scheme on the basis of information collection, summarization and analysis, comprehensively and diversely performs tissue culture experiments, and screens culture mediums at different periods of time, such as primary generation induction → subculture multiplication → rooting culture. The culture of the first generation, the multiplication and the rooting in each period can achieve the best effect, the combination effect is better, and the effect of the flower stick in different stages in the tissue culture propagation process is continuously promoted.
The embodiment of the invention comprises the optimal culture medium for each growth period in the whole tissue culture process of the flower stick, which is obtained through a plurality of experiments in a careful and repeated way, and different types of plant growth agents and different proportions are added according to the requirements of each growth period of the flower stick on the basis of a basic culture medium.
The invention provides a culture medium required by each stage of a tissue culture process of a flower stick, which is characterized by consisting of a primary induction culture medium, a multiplication culture medium and a rooting culture medium.
The optimal culture medium for each phase of the tissue culture of the floral bouquet is screened by a comparative test. The culture medium provided by the invention is suitable for tissue culture of a flower stick, and is particularly suitable for culture of a growing point of the flower stick.
One embodiment of the invention provides a primary induction culture medium of a floral rod: b5+ NAA0.1mg/L +6-BA0.5mg/L + 2.4-D0.1 mg/L + GA30.05-0.1 mg/L + CH 0.5-1 g/L; further preferably B5+ NAA0.1mg/L +6-BA0.5mg/L + 2.4-D0.1 mg/L + GA 30.1 mg/L + CH 0.5 g/L.
The research finds that different hormone combinations can induce the flower stick to differentiate into multiple buds, but the differentiation rates of the combinations are different, B5+ NAA0.1mg/L +6-BA0.5mg/L + 2.4-D0.1 mg/L + GA 30.1 mg/L + CH 0.5g/L is the best induction medium for the growth point of the flower stick, the induction rate is high, the number of newly-increased buds of each explant is large, the differentiation rate is the highest, the differentiation rate can reach 218.75%, and the subculture can be promoted.
One embodiment of the invention provides a subculture medium of a floral rod: modified B5+ NAA 0.3mg/L +6-BA 1mg/L + CH 0.5 g/L; further preferably B5+ NAA 0.3mg/L +6-BA1-2mg/L + CH 0.5-1g/L
The culture medium is the best subculture multiplication culture medium for the flower stick, and not only can produce the most adventitious buds, but also the multiplication times can reach 21.68 times.
One embodiment of the invention provides a rooting culture medium for a flower stick, which comprises the following components in percentage by weight: 1/2MS + IAA0.1mg/L + NAA0.5mg/L + activated carbon 1 g/L.
The research shows that the low salt concentration is favorable for the tissue culture of the anthurium andraeanum, and the culture medium suitable for rooting is 1/2MS, IAA0.1mg/L, NAA0.5mg/L and activated carbon 1 g/L.
In the embodiment of the invention, NAA, 6-BA, 2.4-D, GA3, CH and IAA are short names of naphthylacetic acid, 6-benzylaminopurine, 2.4-dichlorophenoxyacetic acid, gibberellin, hydrolyzed casein and indoleacetic acid respectively within the scope known by the technical personnel in the field.
The optimal culture medium of each growth period in the whole tissue culture process of the flower stick selected by the invention can fully meet the nutritional requirements and growth development of the flower stick at each period, is the key and core for ensuring the successful implementation of the tissue culture of the flower stick and achieving the flow production, and provides powerful support for the large-scale production of the flower stick. Moreover, the stages of primary generation induction, subculture and rooting culture can mutually influence each other, and the selection of the culture medium can continuously promote the floral rod in each stage of tissue culture propagation, so that the yield of industrial floral rod seedling can be greatly improved.
Detailed Description
In order to better explain the present invention and to facilitate the understanding of the technical solutions of the present invention, the present invention is further described in detail below. However, the following examples are only simple examples of the present invention and do not represent or limit the scope of the present invention, which is defined by the claims.
First, material of flower stick explant
The flower stick for experiment is collected from a test nursery of an experiment base of a drought-resistant plant research institute of inner Mongolia Mongolian grass ecological environment (group) GmbH, and is sampled and supplemented at any time according to the test progress and different periods. Collecting growing points of 0.2-0.5cm on tender branches of the flower stick, and cutting under aseptic condition. After washing by running water, sterilizing with 0.1% mercuric chloride solution for 5min, washing with sterile water for 2 times, 5min each time, sucking water on sterile filter paper, inoculating on primary induction culture medium, and establishing test-tube plantlet clone. Selecting cluster buds subcultured for about 10 days, and cutting off the base parts of the cluster buds; the growing point of the cluster bud is taken, the length of the growing point is about 0.2-0.5cm, and the growing point is used as a regenerated explant.
Second, screening statistical method
Survival rate (number of viable explants/total number of inoculated explants) x 100%
Primary shoot induction rate ═ 100% (number of germinated explants/total number of inoculated explants)%
The multiplication factor for subculture (number of cluster buds induced/total number of shoots inoculated) x 100%
Rooting rate ═ 100% (number of explants of differentiated root/total number of explants inoculated)
Example 2 selection of optimal Medium for various stages of tissue culture of floral rods
B5 is used as a basic culture medium, 3.5g/L of carrageenan is used as a curing agent, 40g/L of edible white sugar is used as a carbon source instead of cane sugar, tap water is used instead of distilled water, and cytokinin and auxin in different proportions are respectively added for screening the culture medium.
1. Selection of Primary Induction Medium
Setting two concentration gradients of 0.2mg/L and 0.5mg/L for 6-BA, two concentration gradients of 0.01mg/L and 0.05mg/L for NAA, two concentration gradients of 0.1mg/L and 0.05mg/L for GA3, and two concentration gradients of 0.5mg/L and 1mg/L for CH, performing different combinations, inoculating 2-3 growth points (about 0.2-0.5cm in length) of the flower stick on each bottle, and observing the occurrence of adventitious buds of the growth points on different culture media.
The conditions of primary induction culture are that the culture temperature is 25 +/-1 ℃, the illumination intensity is 1500Lx, and light is supplemented for 4 hours/day at night except for natural illumination in the daytime.
As can be seen from Table 1, 8 hormone combinations can induce the flower stick to differentiate into multiple buds, but the differentiation rates of the combinations are different, namely B5+ NAA0.1mg/L +6-BA0.5mg/L + 2.4-D0.1 mg/L + GA 30.1 mg/L + CH 0.5g/L and B5+ NAA0.1mg/L +6-BA0.5mg/L + 2.4-D0.1 mg/L + GA 30.1 mg/L + CH 1g/L of No. 1 are better induction culture media for the growth point culture medium of the flower stick, the induction rate is high, the number of new buds of each explant is large, the differentiation rate is highest, and the differentiation rate can reach 218.75% and 226.25%.
Table 1: effect of hormone concentration and combination thereof on floral rod growing point multiple bud induction rate
| Treatment of Numbering | Media composition | Number of inoculation (A) | Clumping Number of buds | Cluster bud inducer Conductivity (%) |
| 1 | B5+NAA 0.1mg/L+6-BA 0.5mg/L+2.4-D 0.1mg/ L+GA3 0.1 mg/L+CH 0.5g/L | 80 | 175 | 218.75 |
| 2 | B5+NAA 0.05mg/L+6-BA 0.2mg/L+2.4-D 0.1mg/L+GA3 0.1 mg/L+CH 0.5g/L | 80 | 77 | 96.25 |
| 3 | B5+NAA 0.1mg/L+6-BA 0.2mg/L+2.4-D 0.1mg/ L+GA3 0.1 mg/L+CH 0.5g/L | 80 | 34 | 42.5 |
| 4 | B5+NAA 0.05mg/L+6-BA 0.5mg/L+2.4-D 0.1mg/L+GA3 0.1 mg/L+CH 0.5g/L | 80 | 62 | 77.5 |
| 5 | B5+NAA 0.1mg/L+6-BA 0.5mg/L+2.4-D 0.1mg/ L+GA3 0.05 mg/L+CH 1g/L | 80 | 146 | 182.5 |
| 6 | B5+NAA 0.05mg/L+6-BA 0.2mg/L+2.4-D 0.1mg/L+GA3 0. 05 mg/L+CH 1g/L | 80 | 82 | 102.5 |
| 7 | B5+NAA 0.1mg/L+6-BA 0.2mg/L+2.4-D 0.1mg/ L+GA3 0. 05 mg/L+CH 1g/L | 80 | 113 | 141.25 |
| 8 | B5+NAA 0.05mg/L+6-BA 0.5mg/L+2.4-D 0.1mg/L+GA3 0. 05 mg/L+CH 1g/L | 80 | 37 | 46.25 |
| 9 | B5+NAA 0.1mg/L+6-BA 0.5mg/L+2.4-D 0.1mg/ L+GA3 0.1 mg/L+CH 1g/L | 80 | 181 | 226.25 |
| 10 | B5+NAA 0.05mg/L+6-BA 0.2mg/L+2.4-D 0.1mg/L+GA3 0.1mg/L+CH 1g/L | 80 | 98 | 122.5 |
In this embodiment, the B5 medium is specifically composed of:
macroelements: KNO32500mg/L；MgSO4·7H2O 250mg/L；CaCl2·2H2O 150mg/L；(NH4)2SO4134mg/L；NaH2PO4·H2O 150mg/L
Trace elements: KI 0.75 mg/L; h3BO33mg/L；MnSO4·4H2O 10mg/L；ZnSO4·7H2O 2mg/L；Na2MoO4·2H2O 0.25mg/L；CuSO4·5H2O 0.025mg/L；CoCl2·6H2O 0.025mg/L
Iron salt: FeSO4·7H2O 27.8mg/L；Na2-EDTA 37.3mg/L
Organic matter: inositol 100 mg/L; 1mg/L of nicotinic acid; pyridoxine hydrochloride (vitamin B6)1 mg/L; thiamine hydrochloride (vitamin B1)10 mg/L.
2. Selection of subculture multiplication medium
Respectively inoculating the adventitious buds subjected to induced differentiation to a subculture multiplication medium, marking the serial number of a primary culture medium, setting two concentration gradients of 0.3mg/L and 0.5mg/L for NAA, two concentration gradients of 1mg/L and 2mg/L for 6-BA, two concentration gradients of 0.5g/L and 1g/L for CH, taking eight culture media in total, inoculating 4 single plants in each bottle, observing and recording the differentiation and growth conditions of the regenerated adventitious buds, and counting the multiplication rate of the buds under different hormone combination conditions after 15 days.
The subculture conditions were 25. + -. 1 ℃ for the culture temperature, 2000-3000Lx for the illumination intensity, and 1 hour/day for light supplement at night except for natural illumination in the daytime. After induction culture, 0.2-0.5cm of the growing points of the floral rod cluster buds generated in the induction culture medium are respectively transferred into a subculture multiplication culture medium (see table 2), observation and recording are started when the bud clusters appear on the bud bases of partial materials after 2-3 days, the test results are counted after 15 days, and partial data are shown in table 2 (other data with unobvious characteristics or unobvious effects are not recorded).
Table 2: effect of hormone concentration and combination thereof on adventitious bud proliferation of flower stick
| Treatment of Numbering | Primary culture Base number | Media composition | Proliferation of Multiple of | Growth status of adventitious bud |
| 1 | 1 | Improved B5+ NAA 0.3mg/L +6-BA 1mg/L+CH 0.5g/L | 21.68 | Robust, high emergence of new seedlings, and production of Abundant adventitious buds |
| 2 | 1 | Improved B5+ NAA 0.3mg/L +6-BA 1mg/L+CH 1g/L | 10.49 | Is more robust, has more new seedlings, more vitrification |
| 3 | 1 | Improved B5+ NAA 0.3mg/L +6-BA 2mg/L+CH 0.5g/L | 1.96 | Fine and weak without new emergence of seedlings |
| 4 | 1 | Improved B5+ NAA 0.3mg/L +6-BA 2mg/L+CH 1g/L | 15.64 | Strong and strong, and more new seedlings are produced Less raw glass |
| 5 | 1 | Improved B5+ NAA0.5mg/L +6-BA 1mg/L+CH 0.5g/L | 14.73 | Strong and strong, and more new seedlings are produced Less raw glass |
| 6 | 1 | Improved B5+ NAA0.5mg/L +6-BA 1mg/L+CH 1g/L | 5.84 | Delicate and weak, less new seedlings |
| 7 | 1 | Improved B5+ NAA0.5mg/L +6-BA 2mg/L+CH 0.5g/L | 7.72 | Strong, less new emergence of seedlings and raw Length and slow |
| 8 | 1 | Improved B5+ NAA0.5mg/L +6-BA 2mg/L+CH 1g/L | 11.09 | Strong, more newly emerged seedlings and high yield Less raw vitrification |
| 9 | 9 | Improved B5+ NAA 0.3mg/L +6-BA 1mg/L+CH 0.5g/L | 16.73 | Strong and strong, and more new seedlings are produced More adventitious bud |
| 10 | 9 | Improved B5+ NAA 0.3mg/L +6-BA 2mg/L+CH 1g/L | 12.57 | Is more robust, has more new seedlings, less vitrification |
| 11 | 9 | Improved B5+ NAA0.5mg/L +6-BA 2mg/L+CH 1g/L | 9.72 | Strong, less new seedlings and high yield Less raw vitrification |
Experiments show that the flower stick has a promoting effect on the growth of the cluster buds in the improved B5+ NAA 0.3mg/L +6-BA 1mg/L + CH 0.5g/L subculture, compared with other combinations, the combination is more ideal, the multiplication factor can reach 21.68 times, the number of the cluster buds is large, the seedlings are large, and the effect of transplanting the emergence rate is optimal. However, by comparing the multiplication times of the adventitious buds of different primary culture mediums, the hormone environment of the primary culture medium can be found to influence the adaptability of the adventitious bud in the later propagation stage, and the comprehensive results show that: the combination effect of the primary culture medium B5, the NAA0.1mg/L, the 6-BA0.5mg/L, the 2.4-D0.1 mg/L, the GA 30.1 mg/L and the CH 0.5g/L and the secondary culture medium B5, the NAA 0.3mg/L, the 6-BA 1mg/L and the CH 0.5g/L is the best. In the embodiment, the modified B5 medium is obtained by halving the amounts of macroelements and microelements in the B5 medium, and the basic medium for the floral rod subculture is specifically composed of:
macroelements: KNO31250mg/L；MgSO4·7H2O 125mg/L；CaCl2·2H2O 75mg/L；(NH4)2SO467mg/L； NaH2PO4·H2O 75mg/L
Trace elements: KI 0.375 mg/L; h3BO31.5mg/L；MnSO4·4H2O 5mg/L；ZnSO4·7H2O 1mg/L；Na2MoO4·2H2O 0.125mg/L；CuSO4·5H2O 0.0125mg/L；CoCl2·6H2O 0.0125mg/L
Iron salt: FeSO4·7H2O 27.8mg/L；Na2-EDTA 37.3mg/L
Organic matter: inositol 100 mg/L; 1mg/L of nicotinic acid; pyridoxine hydrochloride (vitamin B6)1 mg/L; thiamine hydrochloride (vitamin B1)10 mg/L.
3. Selection of rooting Medium
Dividing the seedlings with better growth vigor in the subculture medium into single plants, transferring the single plants into a rooting medium, inoculating 4 plants into each bottle, observing and recording the rooting conditions of the seedlings on different culture media, and counting the rooting rate after 10 days.
The rooting culture condition is that the culture temperature is 25 +/-1 ℃, and light is not supplemented at night except for natural illumination in the daytime.
From the test results of 10 days of rooting culture (table 3), the two culture mediums have induction effect on the formation of roots, wherein the No. 2 rooting culture medium has better rooting effect, and the rooting rate reaches 91.56%. The rooting base can be newly added with cluster buds with roots, and the average number can reach 3-4. It can be seen that the low salt concentration is favorable for the growth of the roots of the floral rods, and compared with the influence of the primary culture medium and the secondary culture medium, the seedlings stimulated by high hormone in the primary culture or the secondary culture process are more suitable for the rooting culture medium with high salt concentration. However, the culture medium with low hormone content and low salt concentration is adopted to be matched, so that the better proliferation and rooting effects of the flower stick in primary culture, secondary culture and rooting culture are the results wanted by the technical personnel in the field, the cost can be reduced, and the dependence of the expanding propagation growth of the flower stick on artificial factors can be reduced. Therefore, the culture medium suitable for rooting is 1/2MS + IAA0.1mg/L + NAA0.5mg/L + activated carbon 1 g/L.
Table 3: influence of inorganic salt on seedling rooting
| Processing number | Subculture medium number | Media composition | Rooting percentage (%) | Root growth status |
| 1 | 1 | MS + IAA 0.2mg/L + NAA 1mg/L + active carbon 2g/L | 65.89 | Short, thick and small in quantity |
| 2 | 1 | 1/2MS + IAA0.1mg/L + NAA0.5mg/L + active carbon 1g/L | 91.56 | Long and large in quantity |
| 3 | 9 | MS + IAA 0.2mg/L + NAA 1mg/L + active carbon 2g/L | 74.48 | Short, thick and small in quantity |
| 4 | 9 | 1/2MS + IAA0.1mg/L + NAA0.5mg/L + active carbon 1g/L | 86.74 | Long and large in quantity |
The above examples are intended to illustrate the present invention specifically, but should not be construed as limiting the present invention. Modifications or substitutions to methods, procedures, or conditions of the invention may be made without departing from the spirit and scope of the invention.
Claims (8)
1. A primary induction culture medium of a flower stick is characterized in that the composition of the culture medium is B5+ NAA0.1mg/L +6-BA0.5mg/L + 2.4-D0.1 mg/L + GA30.05-0.1 mg/L + CH 0.5-1 g/L.
2. The subculture medium for a floral rod of claim 1, wherein the primary induction medium is preferably B5+ NAA0.1mg/L +6-BA0.5mg/L + 2.4-D0.1 mg/L + GA 30.1 mg/L + CH 0.5 g/L.
3. The subculture medium for the floral rod is characterized by consisting of improved B5+ NAA 0.3mg/L +6-BA1-2mg/L + CH 0.5-1 g/L.
4. The subculture medium for a floral rod of claim 3, wherein the subculture medium is preferably modified B5+ NAA 0.3mg/L +6-BA 1mg/L + CH 0.5 g/L.
5. The subculture medium for a floral rod of claim 4, wherein the modified B5 medium is obtained by halving the amounts of macroelements and microelements in the B5 medium, and comprises the following components: macroelements: KNO31250mg/L；MgSO4·7H2O 125mg/L；CaCl2·2H2O 75mg/L；(NH4)2SO467mg/L； NaH2PO4·H2O 75mg/L；
Trace elements: KI 0.375 mg/L; h3BO31.5mg/L；MnSO4·4H2O 5mg/L；ZnSO4·7H2O 1mg/L；Na2MoO4·2H2O 0.125mg/L；CuSO4·5H2O 0.0125mg/L；CoCl2·6H2O 0.0125mg/L；
Iron salt: FeSO4·7H2O 27.8mg/L；Na2-EDTA 37.3mg/L；
Organic matter: inositol 100 mg/L; 1mg/L of nicotinic acid; pyridoxine hydrochloride (vitamin B6)1 mg/L; thiamine hydrochloride (vitamin B1)10 mg/L.
6. The rooting culture medium for the flower sticks is characterized by comprising 1/2MS, 0.1mg/L IAA, 0.5mg/L NAA0.5mg/L and 1g/L of activated carbon.
7. A culture medium for tissue culture of a floral rod consists of a primary generation induction culture medium, a secondary generation culture medium and a rooting culture medium; the primary generation induction culture medium is B5+ NAA0.1mg/L +6-BA0.5mg/L + 2.4-D0.1 mg/L + GA30.05-0.1 mg/L + CH 0.5-1g/L, the secondary generation culture medium is improved B5+ NAA 0.3mg/L +6-BA1-2mg/L + CH 0.5-1g/L, and the rooting culture medium is 1/2MS + IAA0.1mg/L + NAA0.5mg/L + active carbon 1 g/L.
8. The culture medium for tissue culture of a floral rod as claimed in claim 7, wherein the composition of the primary culture medium is B5+ NAA0.1mg/L +6-BA0.5mg/L + 2.4-D0.1 mg/L + GA 30.1 mg/L + CH 0.5g/L, and the composition of the secondary culture medium is modified B5+ NAA 0.3mg/L +6-BA 1mg/L + CH 0.5 g/L.
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