A method for cultivating new seaside hibiscus seedlings

Abstract

The present invention provides a method for cultivating new seaside hibiscus seedlings, and belongs to the field of biological agricultural technology. The method for cultivating new seaside hibiscus seedlings provided by the present invention comprises the steps of first subjecting the original seaside hibiscus seedlings to specific aging culture to obtain new seedlings sprouting from the axillary buds of the stems, and then subjecting them to rooting culture to obtain new seaside hibiscus seedlings, which can solve the problem of low seedling formation rate of seaside hibiscus stem segments with buds in tissue culture and limitation of seaside hibiscus propagation in the prior art. The present invention can successfully cultivate and obtain new seaside hibiscus seedlings with excellent traits, and opens up a new way for the protection and propagation of seaside hibiscus germplasm resources.

Description

Technical Field

[0001] The invention relates to the technical field of biological agriculture, and in particular to a method for cultivating new seashore hibiscus seedlings. Background Art

[0002] Hibiscus hamabo is a deciduous shrub or small tree of the genus Hibiscus in the family Malvaceae. Hibiscus hamabo has a dense crown, large golden flowers, a long flowering period, and leaves that turn red in autumn. It is a good ornamental and greening tree species. Hibiscus hamabo has the ecological adaptability of mangrove plants and obvious sea-land transition characteristics. It is called a semi-mangrove tree species. It has a well-developed root system and is extremely resistant to salt and alkali (pH <8.6, salt content <15‰), flooding, and sea breeze. It is an excellent pioneer tree species for embankment forests and coastal trunk forest belts in coastal areas. In addition, Hibiscus hamabo can be used as medicine. The extract in its stem contains anti-tumor ingredients and can also be eaten. Its root bark and leaves can be used to make wine. Anti-photoaging compounds can also be isolated from its body, which has great development potential.

[0003] In the prior art, the main propagation methods of Hibiscus truncatum include sowing, cutting, grafting, etc., which have low seedling yield, slow propagation speed, high price, and cannot meet market demand. Tissue culture technology can be used to obtain a large number of high-quality seedlings with stable traits in a short period of time, reducing production costs. Therefore, it is of great significance to establish a tissue culture rapid propagation system for Hibiscus truncatum to provide a theoretical basis and technical support for the factory production of genetic transformation breeding of Hibiscus truncatum.

[0004] Chinese patent CN117243123A discloses a method for rapid propagation of Hibiscus truncatum through tissue culture, which comprises selecting young stem segments for sterilization, cutting small stem segments with buds and inoculating them into cluster bud induction medium for cluster bud induction and proliferation, and finally selecting seedlings with 4 to 5 true leaves for rooting culture.

[0005] However, in the above scheme, callus is easy to occur in tissue culture of Hibiscus truncatum, and callus restricts the normal growth of the plant. When the stem segments with buds are inoculated into the culture medium, the stem segments are easy to callus and then turn yellow and die. The normal seedling rate of the stem segments with buds is low, which greatly limits the propagation of Hibiscus truncatum. Summary of the invention

[0006] The invention aims to provide a method for cultivating new seaside hibiscus seedlings, which opens up a new way for the protection and expansion of seaside hibiscus germplasm resources.

[0007] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0008] The present invention provides a method for cultivating new seaside hibiscus seedlings, comprising the following steps:

[0009] S1. Inoculate the original seedlings of Hibiscus tiliaceus into the aged culture medium and culture until new seedlings sprout from the axillary buds of the stems;

[0010] S2. Inoculate new seedlings sprouting from the axillary buds of the stems into a rooting medium to obtain new seedlings of Hibiscus syriacus.

[0011] Preferably, the aging medium comprises basal medium, 6-BA, NAA, ABA and ethephon.

[0012] Preferably, the basal medium is MS medium;

[0013] and / or, in the aging medium, the amount of 6-BA added is 0.8 to 1.2 mg / L;

[0014] and / or, in the aging medium, the amount of NAA added is 0.1-0.5 mg / L;

[0015] And / or, in the aging medium, the amount of ABA added is 0.05-0.2 mg / L;

[0016] And / or, in the aging medium, the amount of ethephon added is 0.08-0.12 g / L.

[0017] Preferably, the aging culture medium further comprises activated carbon, and the added amount of the activated carbon is 0.8 to 1.2 g / L.

[0018] Preferably, the rooting medium comprises basal medium, TIBA and NAA.

[0019] Preferably, in the rooting medium, the amount of TIBA added is 0.5-1.5 mg / L;

[0020] And / or, in the rooting medium, the amount of NAA added is 0.05-0.15 mg / L;

[0021] And / or, the basal culture medium is WPM culture medium;

[0022] And / or, the rooting medium further comprises activated carbon and NaCl, the activated carbon is added in an amount of 0.8-1.2 g / L, and the NaCl is added in an amount of 0.08-0.12%.

[0023] Preferably, the culture time in S1 is 3 to 5 months;

[0024] The culture time in S2 is 1 to 3 months.

[0025] Preferably, the Hibiscus sylvestris seedlings are seedlings.

[0026] Preferably, the Hibiscus truncatum seedlings are obtained by the following cultivation method:

[0027] The seaside hibiscus seeds are sterilized and then sown into a germination medium, and the seaside hibiscus seedlings are obtained by germination culture.

[0028] Preferably, the germination culture time is 20 to 40 days;

[0029] The germination medium comprises a basal medium and activated carbon, and the basal medium is an MS medium;

[0030] The added amount of the activated carbon is 0.8-1.2 g / L.

[0031] Beneficial effects of the present invention:

[0032] The method for cultivating new seaside hibiscus seedlings provided by the present invention comprises the steps of first subjecting the original seaside hibiscus seedlings to specific aging culture to obtain new seedlings germinated at the axillary buds of the stems, and then subjecting the seedlings to rooting culture to obtain new seaside hibiscus seedlings. The method can successfully cultivate and obtain new seaside hibiscus seedlings with excellent traits, thus opening up a new way for the protection and expansion of seaside hibiscus germplasm resources. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 Examples of photos of results of culturing in aged culture medium;

[0034] Figure 2 This is a photo of an example of cutting a seedling with a 0.5 cm stem segment;

[0035] Figure 3 This is a photo of the results after aging culture of treatment group 2;

[0036] Figure 4 This is a photo of the results after aging culture of treatment group 3;

[0037] Figure 5 This is a photo of the results after aging culture of treatment group 4;

[0038] Figure 6 This is a photo of the results after aging culture of treatment group 5;

[0039] Figure 7 This is a photo of the results after aging culture of treatment group 6;

[0040] Figure 8 This is a photo of the results after aging culture of treatment group 7;

[0041] Fig. 9 This is a photo of the results after aging culture of treatment group 8;

[0042] Fig.10 This is a photo of the results after aging culture of treatment group 9;

[0043] Fig.11 This is a photo of the results after aging culture of treatment group 10;

[0044] Fig.12 This is a photo of the results of rooting culture of treatment group 11;

[0045] Fig.13 This is a photo of the results of rooting culture of treatment group 12;

[0046] Fig.14 This is a photo of the results of rooting culture of treatment group 13. DETAILED DESCRIPTION

[0047] The present invention provides a method for cultivating new seaside hibiscus seedlings, comprising the following steps:

[0048] S1. Inoculate the original seedlings of Hibiscus tiliaceus into the aged culture medium and culture until new seedlings sprout from the axillary buds of the stems;

[0049] S2. Inoculate new seedlings sprouting from the axillary buds of the stems into a rooting medium to obtain new seedlings of Hibiscus syriacus.

[0050] In the present invention, preferably, the culture time in S1 is 3 to 5 months; the culture time in S2 is 1 to 3 months; preferably, the Hibiscus sabdariffa seedlings are seedlings; preferably, the Hibiscus sabdariffa seedlings are obtained by the following culture method: after sterilizing Hibiscus sabdariffa seeds, sowing them into a germination medium, and germinating and culturing them to obtain Hibiscus sabdariffa seedlings; preferably, the germination and culturing time is 20 to 40 days; the germination medium includes a basal medium and activated carbon, and the basal medium is an MS medium; the amount of activated carbon added is 0.8 to 1.2 g / L.

[0051] In the present invention, preferably, the aging medium comprises a basal medium, 6-BA, NAA, ABA and ethephon; preferably, the basal medium is an MS medium; and / or, in the aging medium, the amount of 6-BA added is 0.8-1.2 mg / L; and / or, in the aging medium, the amount of NAA added is 0.1-0.5 mg / L; and / or, in the aging medium, the amount of ABA added is 0.05-0.2 mg / L; and / or, in the aging medium, the amount of ethephon added is 0.08-0.12 g / L; preferably, the aging medium also comprises activated carbon, and the amount of activated carbon added is 0.8-1.2 g / L. The aging medium can accelerate the growth of seedlings until aging and leaf shedding, such as Figure 1 shown.

[0052] In the present invention, preferably, the rooting medium comprises a basal medium, TIBA and NAA; preferably, in the rooting medium, the addition amount of TIBA is 0.5-1.5 mg / L; and / or, in the rooting medium, the addition amount of NAA is 0.05-0.15 mg / L; and / or, the basal medium is a WPM medium; and / or, the rooting medium further comprises activated carbon and NaCl, the addition amount of the activated carbon is 0.8-1.2 g / L, and the addition amount of the NaCl is 0.08-0.12%.

[0053] In the present invention, by using the above two culture media, new seaside hibiscus seedlings with excellent performance can be successfully cultivated.

[0054] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0055] TDZ: Thidiazuron 96%, 5g / bottle, brand: Mumu Biotechnology.

[0056] NAA: CP / Shanghai 10g.

[0057] ABA: abscisic acid, 30mg / bottle, brand: Mumu Biotechnology.

[0058] 6-BA: 1g / bottle, brand: Sinopharm.

[0059] Ethephon: 100g / bottle, brand: Guoguang.

[0060] Example

[0061] On a clean bench, take the seeds of Hibiscus sylvestris and disinfect them in 75% alcohol for 30 seconds, and in 0.1% mercuric chloride for 5 minutes. After rinsing three times with sterile water, the seeds are evenly sown on the prepared culture medium (MS+1g / L activated carbon) for germination culture.

[0062] After one month, the seeds germinated into seedlings, and then the seedlings were transferred to the prepared aging medium (MS+6-BA1.0mg / L+NAA0.3mg / L+ABA0.1mg / L+ethephon0.1g / L+1g / L activated carbon) for aging culture.

[0063] After 4 months, the plants began to yellow and shed leaves until new seedlings sprouted from the stem axils. Then, the seedlings with 0.5 cm stem segments (such as Figure 2 As shown) were placed in rooting medium (WPM + 1 g / L activated carbon + 0.1% NaCl + TIBA 1.0 mg / L + NAA 0.1 mg / L) for rooting culture.

[0064] Observe the growth of the seedlings after 2 months.

[0065] Experimental example

[0066] In order to study the effects of different types of culture media on the technical solution of the present invention, the following experimental groups were set up:

[0067] Based on the example, other conditions remained unchanged, and only the aged culture medium was replaced with the following numbered culture medium:

[0068] 2: MS+TDZ 1.0mg / L+1g / L activated carbon

[0069] 3: MS+TDZ 1.0mg / L+NAA0.1mg / L+1g / L activated carbon

[0070] 4: MS+TDZ 1.0mg / L+ABA0.1mg / L+1g / L activated carbon

[0071] 5: MS+TDZ 1.0mg / L+NAA 0.3mg / L+ABA0.1mg / L+1g / L activated carbon

[0072] 6: MS+NAA0.3mg / L+1g / L activated carbon

[0073] 7: MS+NAA 0.3mg / L+ABA 0.1mg / L+1g / L activated carbon

[0074] 8: MS+6-BA 0.3mg / L+1g / L activated carbon

[0075] 9: MS+6-BA 1.0mg / L+NAA 0.3mg / L+ABA0.1mg / L+1g / L activated carbon

[0076] 10: MS+6-BA 1.0mg / L+NAA 0.3mg / L+ABA 0.1mg / L+ethephon 0.1g / L+1g / L activated carbon

[0077] Take photos of the aging culture results. Figures 3 to 11 shown.

[0078] From the results, we can see that when NAA is added on the basis of TDZ, the root system of Hibiscus syriacus grows vigorously; after adding ABA, the plant grows densely, the leaves are dark green, and the leaves grow in layers; after adding the combination of ABA and NAA, the root system of the plant is well developed, the leaf spacing becomes larger, and the number of leaves decreases; adding ANN alone or the combination of NAA and ABA is not conducive to plant growth, the seedlings grow weakly, and the plants are short; when ABA is added on the basis of the combination of 6-BA and NAA, the leaves of the plant begin to turn yellow and fall off, and after continuing to add a certain concentration of ethephon, the leaves on the stems of the plant basically fall off, the stems become lignified, and new seedlings sprout from the axillary buds.

[0079] From the above, it can be seen that the combination of 6-BA, NAA and ABA can accelerate the senescence of Hibiscus truncatum plants and promote leaf fall, and ethephon can also promote their senescence and the germination of seedlings from axillary buds of the stems. The solution provided by the present invention can achieve the best effect.

[0080] On the basis of the embodiment, other conditions remain unchanged, only the rooting medium is replaced with the following numbered medium

[0081] 11: WPM+1g / L activated carbon+0.1%NaCl+TIBA 1.0mg / L

[0082] 12: WPM+1g / L activated carbon+0.1%NaCl+TIBA 1.0mg / L+NAA 0.1mg / L

[0083] 13: WPM+1g / L activated carbon+0.1%NaCl+TIBA2.0mg / L+NAA 0.1mg / L

[0084] The rooting situation of the seedlings was statistically analyzed, and the results are shown in Table 1 below:

[0085] Table 1 Effects of different formulations on rooting of Hibiscus truncatum seedlings

[0086] deal with Proliferation coefficient Plant height(cm) Root length (cm) Number of roots 11 2.0 3.2 6.5 2 12 2.0 5.3 9.0 3 13 2.0 4.5 6.2 4

[0087] The photo results of different treatment groups are as follows Figures 12-14 As shown,

[0088] It can be seen from the results that under the three treatments, the seedlings sprouted from the axillary buds of Hibiscus truncatum can take root and grow normally, and two plants can be proliferated at each bud point. After adding NAA and increasing the concentration of TIBA, the number of seedling roots increased, but with the increase of TIBA concentration, the root length and plant height of the seedlings decreased. Comprehensively considering the various indicators, the scheme provided by the present invention can achieve the best effect.

[0089] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A method for cultivating new seaside hibiscus seedlings, characterized in that: The following steps are involved: S1. Inoculate the original seedlings of Hibiscus tiliaceus into the aged culture medium and culture until new seedlings sprout from the axillary buds of the stems; S2, inoculating new seedlings sprouted from the axillary buds of the stems into a rooting medium to obtain new seedlings of Hibiscus syriacus; The seaside hibiscus seedlings are obtained by the following cultivation method: The seaside hibiscus seeds were sterilized and sown into a germination medium to obtain seaside hibiscus seedlings by germination culture; the germination medium was MS + 1g / L activated carbon; The aging medium is MS+6-BA1.0mg / L+NAA0.3mg / L+ABA0.1mg / L+ethephon0.1g / L+1g / L activated carbon; the rooting medium is WPM+1g / L activated carbon+0.1%NaCl+TIBA1.0mg / L+NAA0.1mg / L.

2. The method for cultivating new seaside hibiscus seedlings according to claim 1, wherein: The culture time in S1 is 3 to 5 months; The culture time in S2 is 1 to 3 months.

3. The method for cultivating new seaside hibiscus seedlings according to claim 1 or 2, characterized in that: The original seedlings of Hibiscus tiliaceus are seedlings.

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