CN107646684B

CN107646684B - Cultivation method and application of arundina graminifolia

Info

Publication number: CN107646684B
Application number: CN201710995256.4A
Authority: CN
Inventors: 杨凤玺; 朱根发; 郭阿瑾; 陆楚桥; 金建鹏; 魏永路; 高洁
Original assignee: Environmental Horticulture Institute of Guangdong Academy of Agricultural Sciences
Current assignee: Zhongshan Xijiang Flowers And Trees Research Co ltd
Priority date: 2017-10-23
Filing date: 2017-10-23
Publication date: 2020-01-17
Anticipated expiration: 2037-10-23
Also published as: CN107646684A

Abstract

The invention discloses a cultivation method of arundina graminifolia and application thereof. Picking the bambusa arundinacea capsules growing for 28-30 days after pollination and then performing aseptic seeding; and then, transferring the obtained protocorm to a subculture growth medium, inducing the protocorm to grow and quickly grow seedlings, taking an LED as a tissue culture light source, and carrying out tissue culture by using the following components in a ratio of 2-4: 6-8 parts of red light mixed blue light cultivation; transplanting the obtained aseptic seedlings with the plant height of more than 7cm to a greenhouse for cultivation management. The optimal culture conditions including LED light quality, light intensity and treatment time in the subculture process after the cymbidium arundinaceum seeds germinate are determined, the chlorophyll accumulation, stem thickening and root length extension of plants are effectively promoted, direct transplanting can be realized without a strong seedling rooting process after the subculture, and the production cost and the production period are saved. Especially, the LED promotes the accumulation of flavonoid and phenolic substances of active ingredients in the cymbidium arundinaceum, obviously improves the oxidation resistance of the plant and has important significance for the cultivation of medicinal plants.

Description

Cultivation method and application of arundina graminifolia

Technical Field

The invention relates to the technical field of plant cultivation, in particular to a cultivation method of arundina graminifolia and application thereof.

Background

The Arundina graminifolia (Arundina graminifolia) is a species of cymbidium of orchidaceae, has the effects of regulating and nourishing qi and blood, clearing heat and detoxifying and the like, and is one of main raw materials of the Dai medicine Dai Baijie (Baijie capsules). A large number of researches find that the arundina graminifolia extract has high antioxidant, antitumor, antiviral activity and lipid peroxidation resistance, so that the arundina graminifolia extract can be used for treating diseases such as food poisoning, toxic bacteria poisoning, drug poisoning and the like, wherein the contents of polyphenol, flavone, saponin and the like have obvious dosage effect. In addition, the cymbidium lancifolium flowers are exactly like cattleya hybrida, have large and beautiful flower types, can continuously bloom all the year around, have strong environmental adaptability, and are excellent potted plants and ground cover wild orchids. However, the wild resource of the arundina graminifolia is gradually reduced due to a large amount of artificial digging, and is classified as a national level II protective plant. Therefore, a set of efficient seedling breeding and planting technical system is urgently needed to be established, market demands are met, and popularization and application and industrial development of arundina graminifolia are promoted.

In the plant cultivation management process, the light quality is an important factor influencing the growth and development of plants. Light-emitting diodes (LEDs) are high-quality light sources developed in recent years, which can emit monochromatic light of precise wavelength spectrum, have a narrow wavelength range, an amplitude of not more than 20nm, and can control the light intensity through a control device, thus effectively promoting the photosynthesis of plants, and having a wide application prospect. At present, there are reports that LED light sources are used for tissue culture of plants such as cymbidium hybridum, butterfly orchid, lotus petal orchid, doritaenopsis orchid, chrysanthemum, gerbera jamesonii, calla, white palm, peony, grape, strawberry and the like, and it is found that different light quality matches have larger difference in regulating action of plant growth and development. However, the current research focuses on species with relatively mature scale planting and breeding technologies, and no report is found on light quality regulation in the planting process of the arundina graminifolia.

Disclosure of Invention

In order to overcome the defects of the prior art, the invention aims to provide a cultivation method of arundina graminifolia. The method can effectively promote plant growth, improve medicinal active components, and shorten seedling culture period.

The invention also aims to provide the application of the cultivation method of the arundina graminifolia.

The purpose of the invention is realized by the following technical scheme:

a cultivation method of arundina graminifolia comprises the following steps: sterile seeding, subculture and hardening and transplanting;

(1) and (3) sterile sowing: picking the bambusa arundinacea capsules growing for 28-30 days after pollination, and then performing aseptic seeding; the composition of the culture medium is: the preparation method comprises the following steps of (1) Huabao culture medium, 10-30 g/L of cane sugar, 0.5-1.5 mg/L of 6-benzylpurine (6-BA), 0.1-0.2 mg/L of naphthylacetic acid (NAA), 0.5-1 g/L of activated carbon, 10-30 g/L of banana mud, 6-8 g/L of agar and 5.6-5.8 of pH; culturingConditions are as follows: the light intensity is 20 to 40 mu mo 1m^-2·s^-1Culturing for 25-60 days in a photoperiod of 12-16 h/d;

(2) subculturing: transferring the protocorm obtained in the step (1) to a subculture growth medium, and inducing the protocorm to grow and quickly grow into seedlings under the following culture conditions: LED is used as a tissue culture light source, red light is 620-660 nm, Blue light is 450-480 nm, and the proportion is 2-4: 6-8, light intensity of 30-60 μmo 1. m^-2·s^-1Culturing for 30-60 days in a photoperiod of 12-16 h/d, and increasing the growth speed of plants; the composition of the secondary growth medium is as follows: the preparation method comprises the following steps of (1) Huabao culture medium, 10-30 g/L of cane sugar, 0.5-3 mg/L of 6-benzylpurine (6-BA), 0.2-0.5 mg/L of naphthylacetic acid (NAA), 0.5-1 g/L of activated carbon, 10-30 g/L of banana mud, 6-8 g/L of agar and 5.6-5.8 of pH;

(3) transplanting the test-tube plantlets of the arundina graminifolia: transplanting the aseptic seedlings with the plant height of more than 7cm obtained in the step (2) to a greenhouse for cultivation management; the culture medium comprises: at least one of coconut coir, peat soil, peanut shell, bark and water moss; the cultivation conditions are as follows: keeping the humidity of 60% -80% and 75% for two weeks after transplanting.

The cultivation can be carried out at 20-30 ℃ and 40-60% relative humidity, except for the special specification.

The red light is preferably red light with a wavelength of 630 nm.

The seeding culture medium in the step (1) is preferably Huabao No. 1 culture medium +6-BA0.5mg/L + NAA0.2mg/L + active carbon 1g/L + sucrose 30g/L + banana puree 30g/L + agar 7g/L, and the pH value is 5.6-5.8.

The subculture growth medium in the step (2) is preferably Huabao 1 medium +6-BA1.0mg/L + NAA0.5mg/L + activated carbon 0.5g/L + sucrose 30g/L + banana puree 30g/L + agar 7g/L, and the pH value is 5.6-5.8.

The culture conditions in step (2) are preferably: with LEDs as tissue culture light sources, blue and red light 3: 1 mixing and matching, light intensity of 45 mu mo 1m^-2·s^-1Culturing for 60 days in a photoperiod of 16h/d, wherein the ambient temperature is 24 +/-1 ℃, and the relative humidity is 50-60%.

The new seedlings in the step (2) are preferably the new seedlings with the original bulbodium growing to 2.5-3.5 cm after sowing.

The culture medium in the step (3) is preferably: coconut coir is used as a transplanting substrate, the environmental temperature is 24 +/-1 ℃, and the relative humidity is 60-80%.

The cultivation method of the arundina graminifolia is applied to large-scale cultivation of the arundina graminifolia.

Compared with the prior art, the invention has the following advantages and effects:

(1) the method determines the optimal configuration of different culture media for promoting seed germination in the aseptic seeding process of the arundina graminifolia and promoting the rapid growth of new buds in the subculture process, and effectively promotes the efficient and rapid seedling breeding of the arundina graminifolia.

(2) The optimal culture conditions including LED light quality, light intensity and treatment time in the subculture process after the cymbidium arundinaceum seeds germinate are determined, the chlorophyll accumulation, stem thickening and root length extension of plants are effectively promoted, direct transplanting can be realized without a strong seedling rooting process after the subculture, and the production cost and the production period are saved. Especially, the LED promotes the accumulation of flavonoid and phenolic substances of active ingredients in the cymbidium arundinaceum, obviously improves the oxidation resistance of the plant and has important significance for the cultivation of medicinal plants.

(3) The invention determines the optimal substrate condition for transplanting the aseptic seedlings of the arundo graminifolia, the transplanting survival rate is more than 80 percent, and the industrial production of the arundo graminifolia is effectively promoted.

(4) In the invention, the LED lamp is used as a tissue culture light source, so that the power consumption is saved by 30-50%, the cost is reduced, and the efficiency of large-scale production is improved.

Drawings

FIG. 1 shows the effect of different LED light qualities on chlorophyll fluorescence parameters of tissue culture seedlings of arundina graminifolia.

Detailed Description

The present invention will be described in further detail with reference to examples and drawings, but the present invention is not limited thereto.

Example 1 sterile seeding Medium optimization

Picking the capsules of the chloranthus japonicus which grow for 28-30 days after pollination, sterilizing the capsules for 60s by 70% alcohol, washing the capsules by sterile water, and then using 0.1% HgCl₂Sterilizing the solution for 15min, washing with sterile water for 3-5 times, sucking surface clear water with sterile filter paper, cutting the sterilized capsule with a scalpel, taking out the seed, and spreading the seed in a seeding culture medium sterilized at high temperature and high pressure.

The culture medium takes a Huabao No. 1 culture medium, 7g/L agar and 30g/L sucrose as a control group, 0.5-1.5 mg/L6-benzyl purine (6-BA), 0.1-0.2 mg/L naphthylacetic acid (NAA), 0.5-1 g/L active carbon and 10-30 g/L banana mud as an optimization group, and the specific preparation is shown in Table 1. The pH value is 5.6-5.8.

TABLE 1 seeding Medium formulation

Culture medium formula	6-BA(mg/L)	NAA(mg/L)	Activated carbon (g/L)	Banana mud (g/L)
					CK	0	0	0	0
1	0.5	0.1	0.5	15
					2	0.5	0.2	1	30
3	1.0	0.1	0.5	15
					4	1.0	0.1	1	15
5	1.0	0.2	1	30
					6	1.5	0.1	1	30

10 bottles of seed were seeded for each medium formulation. A Philips (PHILIPS) common white fluorescent lamp tube is used as a light source. The culture conditions are that the photoperiod is 16h/d, the light intensity is 20 mu mo 1m^-2·s^-1The environment temperature is 24 +/-1 ℃, and the relative humidity is 50-60%. After 20 days of continuous culture, the seeds begin to expand, after 40 days of continuous culture, the germination rate (the germination rate is the total number of yellow-white or green expanded and raised seeds/the total number of pollution-free sowed seeds multiplied by 100%) is counted, after 60 days of continuous culture, the height and the leaf number of the new seedlings are counted, and the results are shown in the table2。

TABLE 2 bamboo leaves orchid after 60 days of continuous culture, the height and number of new seedlings are counted

Culture medium formula	Germination rate	Plant height (cm)	Number of blades
				CK	42.98	2.58	2.8
1	62.27	2.74	2.7
				2	72.40	3.61	3.1
3	59.85	2.98	2.8
				4	63.40	3.34	3.2
5	68.83	2.87	3.5
				6	62.19	3.10	3.2

The statistical result (table 2) shows that the germination and growth effects of the formula 2, namely, Huabao No. 1 culture medium +6-BA0.5mg/L + NAA0.2mg/L + activated carbon 1g/L + sucrose 30g/L + banana puree 30g/L + agar 7g/L (pH 5.6-5.8) in different optimized combinations are optimal.

Example 2 Secondary growth Medium composition optimization

And inoculating the new buds in the seeding culture medium into a subculture growth culture medium for rapid growth culture. 10 shoots were inoculated per bottle, 10 bottles were treated each. The culture medium takes a Huabao No. 1 culture medium, 7g/L agar and 30g/L sucrose as a control group, wherein 0.5-3 mg/L6-benzylpurine (6-BA), 0.2-1.0 mg/L naphthylacetic acid (NAA), 0.5-1 g/L activated carbon and 10-30 g/L banana puree are added as an optimization group, and the specific preparation is shown in Table 3. The pH value is 5.6-5.8.

TABLE 3 formulation of subculture growth medium

Culture medium formula	6-BA(mg/L)	NAA(mg/L)	Activated carbon (g/L)	Banana mud (g/L)
					CK	0	0	0	0
1	0.5	0.2	0.5	10
					2	1.0	0.2	1	30
3	1.0	0.5	0.5	30
					4	2.0	0.2	0.5	15
5	2.0	0.5	1	30
					6	3.0	0.2	0.5	20
7	0.5	1.0	1	30
					8	2.0	1.0	1	30

The culture conditions are 16h/d photoperiod and 45 mu mo 1m light intensity^-2·s^-1The environment temperature is 24 +/-1 ℃, and the relative humidity is 50-60%. After the test-tube plantlets are continuously cultured for 60 days, the average plant height of the inoculated buds, the average leaf number and fresh weight of a single plant and the number of newly added buds of the inoculated buds are detected, and specific results are shown in table 4. The plant height was measured with a vernier caliper (to an accuracy of 0.01 mm).

TABLE 4 test tube plantlet after 60d continuous culture test results

Culture medium formula	Plant height	Number of leaves	Root length	Fresh weight (g)
					CK	7.08	5.4	4.9	0.26
1	8.73	6.3	5.5	0.37
					2	9.21	6.1	5.9	0.39
3	8.52	6.2	6.7	0.51
					4	8.16	5.6	5.5	0.42
5	7.98	5.5	6.2	0.45
					6	7.53	5.2	5.1	0.29
7	7.32	5.4	6.4	0.38
					8	7.08	4.9	6.3	0.35

The results show that NAA can promote the growth of the root of the underground part in a certain range. Considering plant height and root length comprehensively, the optimal culture medium formula 3, namely a Huabao No. 1 culture medium +6-BA1.0mg/L + NAA0.5mg/L + activated carbon 0.5g/L + sucrose 30g/L + banana mud 30g/L + agar 7g/L (pH is 5.6-5.8), has a higher growth rate, the plant is thick and strong, the root length is increased by about 50% compared with a control group, and the fresh weight is increased by 2 times higher than that of the control group, so that the optimal successive transfer growth culture medium is selected.

Example 3 Effect of Using culture Medium with LED

1. Subculture growth medium

Huabao 1 medium +6-BA1.0mg/L + NAA0.5mg/L + activated carbon 0.5g/L + sucrose 30g/L + banana puree 30g/L + agar 7g/L (pH 5.6-5.8).

2. Inoculation mode

Inoculating the cymbidium arundinaceum seedlings to a subculture growth medium for multiplication culture after sowing for 60 days. 10 shoots were inoculated per bottle, 10 bottles were treated each.

3. Culture conditions

The light period is 16h/d, and the light intensity is 45 mu mo 1m^-2·s^-1The ambient temperature is 24 +/-1 ℃. In the experiment, 9 treatments were set, specifically, in table 5, White Fluorescent Lamps (WFL) were used as a control, and the LED light source treatments were Warm White (WW), Cool White (CW), far red (DR), red (R), blue (B), and red-blue (R: B: 3: 1, R: B: 2, and R: B: 1: 3, and hereinafter, 3R/1B, 2R/2B, and 1R/3B, respectively). Each treatment was repeated three times, and the light intensity was maintained at 45. mu. mol. m^-2·s^-1The light intensity was measured with an Apogee instrument.

9 light source treatments set forth in Table 5

4. Effect of LEDs on plant growth

Inoculating the new seedling of the cymbidium lophatherum to a new culture medium, continuously culturing for 30 days, and investigating the growth condition of the plant, wherein the statistical items comprise: plant height, root length, stem thickness, leaf length, leaf width; calculating the area of a single leaf according to a formula; weighing fresh weight and dry weight with an electronic balance (deactivating enzyme at 108 deg.C for 15min, oven drying at 80 deg.C for 48 hr to constant weight, measuring soluble sugar content with anthrone colorimetry, measuring soluble protein content with Coomassie brilliant blue method, measuring chlorophyll content with ethanol acetone method, repeating the experiment for 3 times, performing variance analysis with SPSS, and detecting significance of difference with Ducan, P is less than 0.05.

Chlorophyll fluorescence parameters adopt an MINI-PAM-II (WALZ, Germany) ultra-portable modulated chlorophyll fluorescence instrument, and the leaves of the cymbidium arundinaceum seedlings are treated for 30 days without in vitro determination of different LED light qualities. The actual photochemical efficiency Y (II), photochemical quenching coefficient (qp) and non-photochemical quenching coefficient (NPQ) values were directly measured at room temperature, the maximum photochemical efficiency Fv/Fm value was measured at room temperature after the leaves were dark-treated for at least 30 minutes, and each measurement was repeated 3 times. The operation method refers to the instruction of instrument.

TABLE 6 influence of different LED light quality on the growth of tissue culture seedlings of Dianthus hypochondriacus

Light-to-mass ratio

Plant height/mm

Root length/mm

Number of root/number

Thickness of stem/mm

Leaf width/mm

Dry weight/g

Fresh weight/g

T1

56.413^bc

44.1567^a

5^a

3.303^a

5.133^a

0.069^a

0.487^a

T2

58.167^bc

36.987^b

7^ab

3.140^a

4.437^b

0.062^a

0.411^b

T3

61.150^b

36.850^b

5^ab

2.297^bc

3.903^c

0.037^b

0.285^c

T4

58.760^b

31.483^c

4^b

1.787^de

3.530^d

0.033^b

0.246^d

T5

71.943^a

21.138^f

3^b

1.730^c

1.990^g

0.026^b

0.223^d

T6

72.783^a

23.590^f

5^b

1.387^de

2.313^fg

0.030^b

0.234^d

T7

57.083^bc

26.947^de

4^b

2.327^bc

2.537^ef

0.031^b

0.221^d

T8

52.377^c

23.947^ef

3^b

1.927^cd

2.213^fg

0.026^b

0.191^e

CK

59.373^b

28.843^cd

4^b

2.383^b

2.713^e

0.026^b

0.230^d

TABLE 7 influence of different LED photoplasms on pigment content in leaf of tissue-cultured seedling of arundina graminifolia

Light-to-mass ratio	Chlorophyll a/(mg. g)^-1)	Chlorophyll b/(mg. g)^-1)	Total chlorophyll content/(mg. g)^-1)	Chlorophyll a/b
					T1	5.5790^b	2.669^b	8.274^b	2.09^b
T2	7.1460^a	4.328^a	10.7640^a	1.6511^a
					T3	4.2310^c	2.49^c	6.7640^c	1.6991^c
T4	3.8840^e	2.116^e	6^e	1.836^e
					T5	1.9550^g	1.577^g	3.5330^g	1.2397^g
T6	2.4530^b	2.7070^b	5.1520^b	0.906^b
					T7	4.3890^cd	2.433^cd	6.8250^c ^d	1.804^cd
T8	3.4110^f	1.776^f	5.1880^f	1.921^f
					CK	3.9340^d	2.337^d	6.2740^d	1.683^d

TABLE 8 influence of different LED photoplasms on the soluble sugar and protein content of cymbidium lancifolium tissue culture seedlings

Light-to-mass ratio	Soluble sugar/(mg. g)^-1)	Soluble protein/(mg. g)^-1)
			T1	9.5603^f	2.5910^c
T2	9.4373^f	3.9841^a
			T3	10.658^e	3.0260^b
T4	13.407^a	1.7503^e
			T5	12.815^c	1.6680^e
T6	13.055^b	1.5082^f
			T7	11.317^d	2.4130^d
T8	10.879^e	2.3122.3407^d
			CK	11.215^d	2.4972.4787^cd

The influence of different LED light qualities and proportioning treatments on the cymbidium bambusicola tissue culture seedlings is analyzed by measuring plant growth parameters (table 6) such as plant height, root length, stem thickness, leaf area, dry weight, fresh weight and the like, and physiological indexes such as photosynthetic pigment content (table 7), chlorophyll fluorescence parameters (figure 1), soluble sugar content, soluble protein content (table 8) and the like. Compared with the common fluorescence as a light source, the blue light irradiation of 450-480 nm can obviously improve the content of total chlorophyll and soluble protein, promote the growth of root systems, make plants sturdy, increase biomass, but inhibit the overground part of the arundinacea from elongating and growing. While 630nm red light and 735nm far-red light are used as light sources to mainly promote plant height elongation and increase the content of soluble sugar in vivo, but are not beneficial to rooting, obviously inhibit in vivo chlorophyll synthesis and correspondingly reduce biomass. The red and blue light combination can achieve the superposition effect, wherein the ratio of blue light to red light is 3: 1 the best treatment effect of mixing and matching. Compared with common fluorescence, the T2 treatment (3B1R) increases the rooting number of the arundina graminifolia by 75%, the biomass by nearly 80%, the content of soluble protein by 59.5%, the content of total chlorophyll by 71.6%, the values of chlorophyll fluorescence parameters Fv/Fm, Y (II), qp and NPQ are all increased, and the plant photosynthetic capacity is stronger.

5. Effect of LED on active ingredients of arundina graminifolia

The arundina graminifolia has high medicinal value, and the extract has high antioxidant, antitumor, antiviral and lipid peroxidation resisting effects, so that the arundina graminifolia extract can be used for treating diseases such as food poisoning, toxic bacteria poisoning, drug poisoning and the like, wherein the contents of polyphenol, flavone, saponin and the like have obvious dosage effect. Inoculating the new seedling of the arundina graminifolia to a new culture medium, continuously culturing for 30 days, and investigating active ingredients in plants by investigating different LED light quality treatments. Determining the Total Phenol (TP) content of the plant by a Folin-Ciocalteu method; measuring the content of the plant Flavonoid (Flavonoid) by using a NaN02-AI (N03)3 colorimetric method; measuring the superoxide anion scavenging ability of the plants by using a hydroxylamine oxidation method; measuring the scavenging capacity of the hydroxyl radicals of the plants by using a Fenton method; measuring the contents of Catalase (CAT) and Peroxidase (POD) by ultraviolet absorption method; the total antioxidant capacity (T-AOC) of plants was determined by the FRAP method.

TABLE 9 Effect of different LED light quality treatments on Total phenols and Flavonoids in plants

Light-to-mass ratio	Total phenols (mg/g)	Flavonoids (mg/g)
			T1	19.0097^c	25.8747^b
T2	21.1757^a	31.1291^a
			T3	19.2535^b	24.2885^c
T4	16.0270^e	20.5521^d
			T5	12.8461^g	17.7453^e
T6	10.0918^h	10.0242^h
			T7	18.1483^d	15.2380^f
T8	15.8380^f	14.5463^g
			CK	18.0903^d	16.0280^f

TABLE 10 Effect of different LED light quality treatments on antioxidant capacity in plants

As can be seen from the results in tables 9 and 10, the T2 treatment (3B1R) had a significant additive effect on the accumulation of effective active ingredients of arundina graminifolia compared to ordinary fluorescence. The flavonoid is improved by about 2 times, the superoxide anion removal and total oxidation resistance is increased by about 1.5 times, and the flavonoid can be used as a high-quality light source for plant growth and effectively promote the accumulation of medicinal active ingredients.

Example 4 transplantation matrix optimization

Transplanting 3 months old rooting bottle seedlings to a greenhouse, and adding 75% of two layers for shading. Firstly, unscrewing a bottle cap to enable the bottle cap to be breathable, opening the bottle cap after the bottle cap is placed for two weeks to enable the bottle cap to adapt to the external environment, taking out the rooted seedlings from the bottle, cleaning a culture medium at the root, soaking the rooted seedlings in 0.1% of carbendazim for 5-10 min, taking out the rooted seedlings, airing the rooted seedlings, and transferring the rooted seedlings into a matrix. And (5) thoroughly watering with a sprayer to ensure that the seedlings are attached to the matrix. And then keeping the environmental humidity at about 80% for two weeks, and keeping the humidity at 50-60% after the seedlings survive.

And (5) measuring the growth index after transplanting the aseptic seedlings for 2 months. The survey items are: survival rate, average plant height, single plant leaf number, newly increased bud number and fresh weight. The plant height was measured with a vernier caliper (to an accuracy of 0.01 mm). The results are shown in Table 11.

TABLE 11 determination of growth index 2 months after transplanting of aseptic seedlings

Substrate	Plant height (cm)	Number of leaves	Fresh weight (g)	Survival rate (%)
					T1	13.57	6.9	0.97	86.5
T2	11.71	6.1	0.84	70.5
					T3	12.52	6.3	1.11	80.2
T4	13.09	6.6	1.03	78.6
					T5	13.78	6.5	1.16	73.2
T6	10.89	6.1	1.0 9	70
					T7	11.08	6.9	1.04	71.9
T8	14.31	7.1	1.17	73.4
					T9	12.53	7.0	1.13	82.3

Therefore, the arundina graminifolia has strong adaptability, the survival rate of the arundina graminifolia in different matrixes is more than 70 percent, but the growth conditions of different treatments have larger difference. The test result shows that the survival rate of the transplanted seedlings is the highest after the coconut coir is used as the matrix, the growth vigor of the seedlings is better after the water moss is transplanted to live, but the survival rate is lower than that of the coconut coir used as the matrix, and the survival rate is probably related to the water retention capacity. Comprehensively comparing the coconut chaff used as the substrate most suitable for the growth of the cymbidium arundinaceum test-tube plantlet. It can be seen that, different from other tropical orchid with aerial roots, the coco coir cultivation with good air permeability and water retention is beneficial to the survival of aseptic seedlings and the increase of the growth rate of the aseptic seedlings for the cymbidium bambusoides with the terrestrial property.

The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.

Claims

1. A cultivation method of arundina graminifolia is characterized by comprising the following steps: sterile seeding, subculture and hardening and transplanting;

(1) and (3) sterile sowing: picking the bambusa arundinacea capsules growing for 28-30 days after pollination, and then performing aseptic seeding; the composition of the culture medium is: the preparation method comprises the following steps of (1) a Huabao 1 culture medium, 10-30 g/L of cane sugar, 0.5-1.5 mg/L of 6-BA, 0.1-0.2 mg/L of NAA, 0.5-1 g/L of active carbon, 10-30 g/L of banana mud and 6-8 g/L of agar, wherein the pH is 5.6-5.8; the culture conditions are as follows: the light intensity is 20-40 mu mo 1.m^-2•s^-1Culturing for 25-60 days in a photoperiod of 12-16 h/d;

(2) subculturing: transferring the protocorm obtained in the step (1) to a subculture growth medium, and inducing the protocorm to grow and quickly grow into seedlings under the following culture conditions: an LED is used as a tissue culture light source, and Blue light Blue 450-480 nm and red light 630nm are mixed according to the proportion of 3: 1 mixing and matching, light intensity of 45 mu mo 1. m^-2•s^-1Culturing for 60 days in a photoperiod of 16h/d, wherein the ambient temperature is 24 +/-1 ℃, and the relative humidity is 50-60%; the composition of the secondary growth medium is as follows: the preparation method comprises the following steps of (1) Huabao culture medium +6-BA0.5mg/L + NAA0.2mg/L + activated carbon 1g/L + sucrose 30g/L + banana puree 30g/L + agar 7g/L, and the pH is = 5.6-5.8;

2. The cultivation method of arundina graminifolia according to claim 1, characterized in that:

the culturing in the steps (1), (2) and (3) is carried out under the conditions of 20-30 ℃ and 40-60% of relative humidity except for special instructions.

3. The cultivation method of arundina graminifolia according to claim 1, characterized in that:

the sowing culture medium in the step (1) is a Huabao No. 1 culture medium +6-BA0.5mg/L + NAA0.2mg/L + active carbon 1g/L + sucrose 30g/L + banana puree 30g/L + agar 7g/L, and the pH = 5.6-5.8.

4. The cultivation method of arundina graminifolia according to claim 1, characterized in that:

the protocorm in the step (2) is grown to 2.5-3.5 cm after being sown.

5. The cultivation method of arundina graminifolia according to claim 1, characterized in that:

the culture medium in the step (3) is as follows: coconut coir is used as a transplanting substrate, the environmental temperature is 24 +/-1 ℃, and the relative humidity is 60-80%.

6. The application of the arundina graminifolia culture method of any one of claims 1 to 5 in large-scale cultivation of arundina graminifolia with high medicinal active ingredients is characterized in that: the medicinal active ingredients are flavonoids and phenols.