CN111213587B - Tissue culture rapid propagation method of rose apple - Google Patents
Tissue culture rapid propagation method of rose apple 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
- A01H4/008—Methods for regeneration to complete plants
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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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- 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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
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- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
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Abstract
The invention discloses a tissue culture rapid propagation method of rose apple, comprising the steps of explant disinfection, axillary bud induction, proliferation culture and rooting culture, wherein the axillary bud induction is to inoculate the disinfected explant into an axillary bud induction culture medium for culture for 18-22 d to obtain sterile buds; the axillary bud induction culture medium comprises the following components: the improved WPM, 6-BA 0.01-0.03 mg/L, NAA 0.01-0.02 mg/L, L-leucine 1.0-2.0 mg/L, tomato juice 40-60 g/L. The invention adopts the improved WPM culture medium combined with leucine and tomato juice to greatly reduce the hormone dosage in the axillary bud induction period, shorten the axillary bud induction period and reduce the cost.
Description
Technical Field
The invention belongs to a plant tissue culture propagation method, and particularly relates to a tissue culture rapid propagation method of rose apple.
Background
Syzygium jambos (Xanthostemon chrysanthus F. Muell. ex Benth) is an evergreen small arbor of the genus Bupleurum of the family Myrtaceae (Myrtaceae), native Australia, one of representative plants of tropical rainforest of Australia, is a flower of Karens city (Cairns) of Queensland, and is introduced to the places of Fujian, Guangdong and the like in recent years. The leaf of the golden rose apple is opposite, intergrown or fasciculate, needle-shaped, complete, leathery, smooth in leaf surface, has guava smell after being kneaded, and the new leaf is red. The inflorescence is spherical, the axils of the apical or branch tip leaves bloom, the color is yellow green at the initial stage of blooming, the color is changed into yellow along with time, and the color is golden yellow when the inflorescence withers; although the flower is a flower, the petals of the flower are degenerated, only obvious circular sepals and stamens exist, the stamens and the stamens extend out, the filaments are radiated, the flower is unique all the year around, the full-bloom period is from 11 months to 2 months next year, the fruit is a capsule, and the fruit has a persistent pistil. The golden cattail has tall and straight plant, strong growth vigor, bright leaf color, large flower quantity and golden yellow tree when flowering in winter, and is extremely spectacular, thereby creating a beautiful landscape in winter.
The tissue culture seedling obtained by the method for tissue culture propagation of the rose apple has the characteristics of complete root system, large growth amount, no influence of seasons on seedling culture, high propagation rate and the like, and is more suitable for large-scale and standardized seedling culture in production. The tissue culture and rapid propagation technology of the golden syzygium jambos (Chenqiming, forestry investigation and design, 2019, 02) discloses that axillary bud induction, proliferation, rooting and seedling hardening transplantation experiments are carried out by taking the golden syzygium stems as explants, and the results show that: the more suitable axillary bud induction culture medium for the stem section with bud of the golden syzygium jambos is as follows: 1.0mg/L of MS +6-BA, 0.5mg/L of NAA, 30g/L of white sugar and 5.8g/L of carrageenan, wherein the inductivity reaches 90.78 percent, and the bud induction time is 30 days; the optimal formula of the subculture multiplication medium is as follows: MS +6-BA 0.6mg/L + NAA0.3mg/L + IBA0.1mg/L + riboflavin 1g/L + white sugar 30g/L + carrageenan 5.8g/L, and the subculture multiplication coefficient reaches 3.58 times; the optimal root induction culture medium formula is as follows: MS, IBA0.6mg/L, ABT 0.2mg/L, white sugar 30g/L, carrageenan 5.8g/L and active carbon 0.2g/L, wherein the rooting rate can reach 94.5%; peat soil and fermented fir bark are used as transplanting substrates, and the average transplanting survival rate is 92.0%. However, the method has the advantages of large dosage of auxin and mitogen and long bud induction period.
Disclosure of Invention
The invention aims to provide a tissue culture rapid propagation method of rose apple with small dosage of phytohormone and short bud induction period.
The technical scheme provided by the invention is a tissue culture rapid propagation method of rose apple, which comprises axillary bud induction, wherein a sterilized explant is inoculated into an axillary bud induction culture medium to be cultured for 18-22 d to obtain a sterile bud; the axillary bud induction culture medium comprises the following components: improved WPM, 6-BA 0.01-0.03 mg/L, NAA 0.01-0.02 mg/L, L-leucine 1.0-2.0 mg/L, tomato juice 40-60 g/L;
the formula of the improved WPM culture medium is as follows: NH (NH) 4 NO 3 400mg/L、CaCl 2 ·2H 2 O 96mg/L、MgSO4·7H 2 0 370mg/L、KH 2 PO 4 170mg/L、Ca(NO 3 ) 2 ·4H 2 O 360mg/L、KI 0.83mg/L、H 3 BO 3 6.2mg/L、MnSO 4 ·H 2 O 22.4mg/L、ZnSO4·7H 2 O 8.6mg/L、Na 2 MoO 4 ·2H 2 O 0.25mg/L、CuSO 4 ·5H 2 O 0.25mg/L、Na 2 ·EDTA37.3mg/L、FeSO4·7H 2 27.8mg/L of O, 100mg/L of inositol, 0.5mg/L of nicotinic acid, 1.0mg/L of pyridoxine hydrochloride (B6), 2.0mg/L of thiamine hydrochloride (B1), 2.0mg/L of glycine, 20.0g/L of sucrose, 6.0g/L of agar and 5.0-5.5 of pH.
The improved WPM culture medium is adopted in the axillary bud induction culture medium, and the L-leucine and the tomato juice are compounded, so that a good axillary bud induction effect can be ensured and the axillary bud induction period can be shortened under the condition that the dosage of hormones is greatly reduced. The bud inductivity is more than 90%, and the sterile bud is about 2cm long.
The tomato juice can be prepared according to a conventional method, and preferably, the preparation method of the tomato juice comprises the following steps: the tomato juice is prepared by cutting tomatoes, adding the cut tomatoes into water with the weight being 10-30 times of that of the tomatoes, heating the tomato to boiling and maintaining the boiling for 3-5 mm, and filtering to obtain filtrate, namely the tomato juice.
The axillary bud induction conditions are as follows: culturing for 6-8 days under the conditions that the illumination intensity is 0-500 lx, the temperature is 22 +/-2 ℃ and the humidity is 60-70%; and then keeping the temperature and the humidity unchanged, improving the illumination intensity to 1500-2000 lx, keeping the illumination time to 12-14 h/d, and continuing culturing for 12-14 d.
The enrichment culture is to cut the aseptic buds into small sections according to internodes and inoculate the small sections on an enrichment culture medium for culture for 25-30 d to obtain cluster buds; the formula of the proliferation culture medium is as follows: the improved WPM, 6-BA 0.03-0.05 mg/L, NAA 0.01-0.02 mg/L, L-leucine 1.0-2.0 mg/L, tomato juice 40-60 g/L.
The improved WPM, L-leucine and tomato juice are adopted in the proliferation culture medium, so that a good bud proliferation effect can be achieved under the condition that a very small amount of hormone is added, and the proliferation coefficient reaches more than 3.58.
The proliferation culture conditions are as follows: the temperature is 26 +/-2 ℃, the illumination is 1000-2000 lx, the illumination time is 14-16 h/d, and the humidity is 70-80%.
The rooting culture is to cut off single cluster buds, inoculate the cut cluster buds to a WPM culture medium without any hormone for culture for 7-10 days, and then transfer the cultured cluster buds to a rooting culture medium for culture for 10-14 days to obtain rooted seedlings; the formula of the rooting culture medium is as follows: 3/4WPM and NAA 0.5-1 mg/L.
In the process of rooting culture, WPM without any hormone is firstly used for culture and then transferred to a rooting culture medium for culture, and the rooting rate can reach more than 96%.
The WPM formula is as follows: NH 4 NO 3 400mg/L、Ca(NO 3 ) 2 ·4H 2 O 556mg/L、K 2 SO 4 99Omg/L、CaCl 2 ·2H 2 O 96mg/L、KH 2 PO 4 170mg/L、Na 2 MoO 4 ·2H 2 O 0.25mg/L、MgSO4·7H 2 0 370mg/L、MnSO 4 ·H 2 O 22.4mg/L、ZnSO4·7H 2 O 8.6mg/L、CuSO 4 ·5H 2 O 0.25mg/L、FeSO4·7H 2 O 27.8mg/L、Na 2 EDTA37.3mg/L, inositol 100mg/L, thiamine hydrochloride (B1)1.0mg/L, nicotinic acid 0.5mg/L, pyridoxine hydrochloride (B6)0.5mg/L, glycine 2.0mg/L, sucrose 20.0g/L, agar 6.0g/L, pH 5.0-5.5.
The rooting culture conditions are as follows: the temperature is 25 +/-2 ℃, the illumination intensity is 1000-1500 lx, the illumination time is 10-12 h/d, and the humidity is 70-80%.
Compared with the prior art, the invention has the following advantages:
1) the axillary bud induction culture medium disclosed by the invention takes the improved WPM culture medium as a basic culture medium, and the L-leucine and the tomato juice are added, so that the conventional axillary bud induction period is shortened to 18-22 days from 30 days under the condition of greatly reducing the dosage of hormone, and the axillary bud induction rate is ensured to be higher than 90%.
2) The improved WPM culture medium is adopted in the proliferation culture medium, and the L-leucine and the tomato juice are added, so that a good proliferation effect can be achieved under the condition that the dosage of hormones is greatly reduced, and the proliferation coefficient is over 3.58.
Detailed Description
The following examples further illustrate the invention but are not intended to limit it.
Example 1
(1) Selection of explants: in early spring, cutting a semi-lignified new shoot with a latent bud which grows in the current year from a flowering mother plant of the rose apple as an explant; diluting the whole plant by 2000 times of triazolone missible oil with the effective content of 20 percent one day before pruning, and spraying the whole plant for 1 time.
(2) Explant disinfection: cutting off leaves on the branches, washing for 10-15 min by using tap water, then washing by using a soft-bristle toothbrush in combination with a proper amount of detergent, and brushing the branches for 2-3 times during washing; then washing with sterilized water for 4-5 times, cutting the branches into stem sections with 1-2 axillary buds, transferring to a superclean bench, placing into a sterilized container, and adding 0.1% HgCl 2 Soaking for 8-12 min, and finally washing with sterilized water for 4 times for later use.
(3) Axillary bud induction: transferring the sterilized stem segments into an inoculation dish by using a pair of tweezers, cutting 2-3 mm lower morphological ends of the stem segments into inclined planes, keeping morphological upper ends, inserting the inclined planes into an axillary bud induction culture medium, inoculating 3-5 stem segments into each bottle, culturing for 6 days under the conditions that the illumination intensity is 0lx, the temperature is 20 ℃ and the humidity is 60%, then increasing the illumination intensity to 1500lx under the conditions that the temperature and the humidity are not changed, and continuously culturing for 12 days under the illumination time of 12h/d to obtain sterile buds.
The axillary bud induction culture medium comprises the following components: improved WPM, 6-BA0.01 mg/L, NAA 0.01.01 mg/L, L-leucine 1.0mg/L, tomato juice 40 g/L;
the preparation method of the tomato juice comprises the following steps: crushing tomato, adding 10 times of water, heating to boil and maintaining for 3mm, and filtering to obtain filtrate.
The formula of the improved WPM culture medium is as follows: NH 4 NO 3 400mg/L、CaCl 2 ·2H 2 O 96mg/L、MgSO4·7H 2 0 370mg/L、KH 2 PO 4 170mg/L、Ca(NO 3 ) 2 ·4H 2 O 360mg/L、KI 0.83mg/L、H 3 BO 3 6.2mg/L、MnSO 4 ·H 2 O 22.4mg/L、ZnSO4·7H 2 O 8.6mg/L、Na 2 MoO 4 ·2H 2 O 0.25mg/L、CuSO 4 ·5H 2 O 0.25mg/L、Na 2 ·EDTA 37.3mg/L、FeSO4·7H 2 27.8mg/L of O, 100mg/L of inositol, 0.5mg/L of nicotinic acid, 1.0mg/L of pyridoxine hydrochloride (B6), 2.0mg/L of thiamine hydrochloride (B1), 2.0mg/L of glycine, 20.0g/L of sucrose, 6.0g/L of agar and 5.0-5.5 of pH.
Comparative example 1
Example 1 was repeated except that:
the formula of the axillary bud induction culture medium in the step (3) is as follows: improved WPM, 6-BA0.01 mg/L, NAA 0.01.01 mg/L, tomato juice 40 g/L.
Comparative example 2
Example 1 was repeated except that:
the formula of the axillary bud induction culture medium in the step (3) is as follows: the WPM and 6-BA are improved, wherein the proportion of the WPM and the 6-BA is 0.01mg/L, NAA 0.01.01 mg/L, L-leucine is 1.0 mg/L.
Comparative example 3
Example 1 was repeated except that:
the axillary bud induction culture medium in the step (3) has the formula: WPM, 6-BA0.01 mg/L, NAA 0.01.01 mg/L, L-leucine 1.0mg/L, tomato juice 40 g/L.
Comparative example 4
Example 1 was repeated, except that the axillary bud induction medium in step (3) was the axillary bud induction medium of "rapid tissue culture and propagation technique of rose apple (aged, forestry investigation design" 2019, stage 02), and the formula was: MS +6-BA 1.0mg/L + NAA0.5 mg/L + white sugar 30g/L + carrageenan 5.8 g/L.
Example 2
The sterile bud obtained by the method in example 1 is tested according to proliferation, rooting, hardening and transplanting in the rapid propagation technology of tissue culture of rose apple (aged brightness, forestry investigation and design, 2019, 02), and specifically comprises the following steps:
(4) and (3) proliferation culture: cutting the aseptic buds into small segments according to internodes, obliquely inserting the small segments into a multiplication culture medium, and culturing for 25d to obtain cluster buds. The formula of the proliferation culture medium is as follows: MS +6-BA 0.6mg/L + NAA0.3mg/L + IBA0.1mg/L + riboflavin 1g/L + white sugar 30g/L + carrageenan 5.8 g/L. The proliferation culture conditions are as follows: the temperature is 24 ℃, the illumination is 1000lx, the illumination time is 14h/d, and the humidity is 70 percent.
(5) Rooting culture: shearing off the cluster bud single plant, inoculating the cluster bud single plant to a WPM culture medium, culturing for 7d, and then transferring to a rooting culture medium, culturing for 10d to obtain a rooted seedling. The formula of the rooting culture medium is as follows: MS, IBA0.6mg/L, ABT 0.2mg/L, white sugar 30g/L, carrageenan 5.8g/L and active carbon 0.2 g/L. The rooting culture conditions are as follows: the temperature is 23 ℃, the illumination intensity is 1000lx, the illumination time is 10h/d, and the humidity is 70 percent.
(6) Hardening and transplanting seedlings: hardening the rooted seedlings obtained in the step (5) in a seedling hardening chamber for 2d, then removing the cover and hardening the seedlings for 1d, and hardening the roots of the rooted seedlings by using KMnO with the mass fraction of 0.05 percent 4 Soaking in solution for 1min, transplanting to seedbed, taking peat soil and fermented fir bark as transplanting matrix, and thoroughly spraying with water before use. Keeping the leaf surface humidity after transplanting.
Example 3
The sterile shoots obtained as described in example 1 were used for further cultivation.
(4) And (3) proliferation culture: cutting the aseptic buds into small segments according to internodes, obliquely inserting the small segments into a multiplication culture medium, and culturing for 25d to obtain cluster buds. The formula of the proliferation culture medium is as follows: improved WPM, 6-BA0.03mg/L, NAA0.01mg/L, L-leucine 1.0mg/L, and tomato juice 40 g/L. The proliferation culture conditions are as follows: the temperature is 24 ℃, the illumination is 1000lx, the illumination time is 14h/d, and the humidity is 70 percent.
(5) Rooting culture: shearing off the cluster bud single plant, inoculating the cluster bud single plant to a WPM culture medium, culturing for 7d, and then transferring to a rooting culture medium, culturing for 10d to obtain a rooted seedling. The formula of the rooting culture medium is as follows: 3/4WPM and NAA are modified at 0.5-1.0 mg/L. The rooting culture conditions are as follows: the temperature is 23 ℃, the illumination intensity is 1000lx, the illumination time is 10h/d, and the humidity is 70 percent.
(6) Hardening and transplanting seedlings: hardening the rooted seedlings obtained in the step (5) in a seedling hardening chamber for 2d, then removing the cover and hardening the seedlings for 1d, and hardening the roots of the rooted seedlings by using KMnO with the mass fraction of 0.05 percent 4 Soaking the solution for 1min, and then transplanting the solution to a seedbed, wherein the matrix is prepared from the following components in percentage by mass: 2: 1 peat soil, perlite and vermiculite, and before using the seedbed, water is usedAnd (6) thoroughly leaching. Keeping the leaf surface humidity after transplanting.
Comparative example 5
Example 3 was repeated, except that the proliferation medium in step (4) was: improved WPM, 6-BA0.03mg/L, NAA0.01mg/L, and tomato juice 40 g/L.
Comparative example 6
Example 3 was repeated, except that the proliferation medium in step (4) was: improved WPM, 6-BA0.03mg/L, NAA 0.01.01 mg/L, L-leucine 1 mg/L.
Comparative example 7
Example 3 was repeated, except that the proliferation medium in step (4) was: WPM, 6-BA0.03mg/L, NAA0.01mg/L, L-leucine 1mg/L, and tomato juice 40 g/L.
Comparative example 8
Example 3 was repeated except that the rooting medium of step (5) was the rooting medium of "tissue culture and rapid propagation of rose apple (chenqiming, forestry investigation design" 2019, stage 02) and the formulation thereof was: MS + IBA0.6mg/L + ABT 0.2mg/L + white sugar 30g/L + carrageenan 5.8g/L + active carbon 0.2 g/L.
Example 4
(1) And (2) same as example 3.
(3) Axillary bud induction: transferring the sterilized stem segments into an inoculation dish by using a pair of tweezers, cutting 2-3 mm lower morphological ends of the stem segments into inclined planes, keeping morphological upper ends, inserting the inclined planes into an axillary bud induction culture medium, inoculating 3-5 stem segments into each bottle, culturing for 8 days under the conditions of illumination intensity of 500lx, temperature of 24 ℃ and humidity of 70%, then increasing the illumination intensity to 2000lx under the conditions of unchanged temperature and humidity, and continuously culturing for 14 days under the illumination time of 14h/d to obtain sterile buds.
The axillary bud induction culture medium comprises the following components: improved WPM, 6-BA0.03mg/L, NAA0.02mg/L, L-leucine 2.0mg/L, tomato juice 60 g/L;
the preparation method of the tomato juice comprises the following steps: crushing tomato, adding 30 times of water, heating to boil and maintaining for 5 mm, and filtering to obtain filtrate.
(4) And (3) proliferation culture: cutting the aseptic buds into small segments according to internodes, obliquely inserting the small segments into a multiplication culture medium, and culturing for 30d to obtain cluster buds. The formula of the proliferation culture medium is as follows: modified WPM, 6-BA0.05mg/L, NAA0.02mg/L, L-leucine 2.0mg/L, tomato juice 60 g/L. The proliferation culture conditions are as follows: the temperature is 28 ℃, the illumination is 2000lx, the illumination time is 16h/d, and the humidity is 80 percent.
(5) Rooting culture: shearing off the cluster bud single plant, inoculating the cluster bud single plant to a WPM culture medium, culturing for 10d, and then transferring to a rooting culture medium, culturing for 14d to obtain a rooted seedling. The formula of the rooting culture medium is as follows: 3/4WPM was modified to 1.0 mg/L. The rooting culture conditions are as follows: the temperature is 27 ℃, the illumination intensity is 1500lx, the illumination time is 12h/d, and the humidity is 80 percent.
(6) Hardening and transplanting seedlings: hardening the rooted seedlings obtained in the step (5) in a seedling hardening chamber for 3d, then, covering and hardening the seedlings for 3d, soaking the roots of the hardened rooted seedlings in a carbendazim solution with the mass ratio of 800 times for 3min, and then, transplanting the roots onto a seedbed, wherein the matrix comprises the following components in percentage by mass: 2: 1, and before the seedbed is used, the seedbed is thoroughly drenched with water. Keeping the leaf surface humidity after transplanting.
Growth parameters in the cultivation processes of examples 1-4 and comparative examples 1-7 are shown in the following table:
Claims (7)
1. a tissue culture rapid propagation method of rose apple comprises the steps of explant disinfection, axillary bud induction, proliferation culture and rooting culture, and is characterized in that: in the axillary bud induction, the sterilized explant is inoculated into an axillary bud induction culture medium to be cultured for 18-22 d to obtain a sterile bud; the axillary bud induction culture medium comprises the following components: improved WPM, 6-BA 0.01-0.02 mg/L, NAA 0.01-0.02 mg/L, L-leucine 1.0-2.0 mg/L and tomato juice 40-60 g/L;
the formula of the improved WPM culture medium is as follows: NH (NH) 4 NO 3 400 mg/L、CaCl 2 ·2H 2 O 96 mg/L、MgSO 4 ·7H 2 O 370 mg/L、KH 2 PO 4 170 mg/L、Ca(NO 3 ) 2 ·4H 2 O 360 mg/L、KI 0.83 mg/L、H 3 BO 3 6.2 mg/L、MnSO 4 ·H 2 O 22.4 mg/L、ZnSO 4 ·7H 2 O 8.6 mg/L、Na 2 MoO 4 ·2H 2 O 0.25 mg/L、CuSO 4 ·5H 2 O 0.25 mg/L、Na 2 ·EDTA 37.3 mg/L、FeSO 4 ·7H 2 27.8mg/L of O, 100mg/L of inositol, 0.5mg/L of nicotinic acid, 1.0mg/L of pyridoxine hydrochloride, 2.0mg/L of thiamine hydrochloride, 2.0mg/L of glycine, 20.0g/L of sucrose, 6.0g/L of agar and pH of 5.0-5.5.
2. The tissue culture rapid propagation method of rose apple according to claim 1, characterized in that: the preparation method of the tomato juice comprises the following steps: crushing tomatoes, adding water with the weight 10-30 times of that of the tomatoes, heating to boil, maintaining for 3-5 min, and filtering to obtain filtrate, namely tomato juice.
3. The tissue culture rapid propagation method of rose apple according to claim 1, characterized in that: the axillary bud induction conditions are as follows: culturing for 6-8 d under the conditions that the illumination intensity is 0-500 lx, the temperature is 22 +/-2 ℃ and the humidity is 60-70%; and then keeping the temperature and the humidity unchanged, increasing the illumination intensity to 1500-2000 lx, keeping the illumination time to 12-14 h/d, and continuing culturing for 12-14 d.
4. The tissue culture rapid propagation method of Eugenia jambolana according to any one of claims 1 to 3, which is characterized in that: the enrichment culture is to cut the aseptic buds into small sections according to internodes and inoculate the small sections on an enrichment culture medium for culture for 25-30 d to obtain cluster buds; the formula of the proliferation culture medium is as follows: the improved WPM, 6-BA 0.03-0.05 mg/L, NAA 0.01-0.02 mg/L, L-leucine 1.0-2.0 mg/L, tomato juice 40-60 g/L.
5. The tissue culture rapid propagation method of Eugenia jambolana according to claim 4, which comprises: the proliferation culture conditions are as follows: the temperature is 26 +/-2 ℃, the illumination is 1000-2000 lx, the illumination time is 14-16 h/d, and the humidity is 70-80%.
6. The tissue culture rapid propagation method of rose apple according to any one of claims 1 to 3, characterized in that: the rooting culture is to cut off single cluster buds, inoculate the single cluster buds to a WPM culture medium for culture for 7-10 days, and then transfer the single cluster buds to a rooting culture medium for culture for 10-14 days to obtain rooted seedlings; the formula of the rooting culture medium is as follows: 3/4WPM and NAA 0.5-1 mg/L.
7. The tissue culture rapid propagation method of Eugenia jambolana according to claim 6, which comprises: the rooting culture conditions are as follows: the temperature is 25 +/-2 ℃, the illumination intensity is 1000-1500 lx, the illumination time is 10-12 h/d, and the humidity is 70-80%.
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