Growth regulator for rapid propagation of sweet potato stems as well as use method and application of growth regulator

Abstract

The invention discloses a growth regulator for rapid propagation of sweet potato stems and a use method and application thereof, and belongs to the technical field of plant tissue culture. In order to solve the technical problems that in the prior art, the period of rapid propagation of sweet potatoes is long, root growth is slow, and large-scale seedling growing requirements cannot be met, the invention provides a growth regulator for rapid propagation of sweet potato stems, and the growth regulator contains allantoin with the concentration of 250 mg / L. The allantoin-containing growth regulator provided by the invention can obviously promote the average growth amount of the single leaf area, the root length elongation and the root fresh weight of the sweet potato virus-free seedlings; the growth regulator independently treats the sweet potato stem segments, compared with NAA or the growth regulator and NAA cooperatively treat the sweet potato stem segments, the growth regulator has the better rapid propagation effect, growth of the roots of the sweet potato virus-free seedlings is remarkably promoted, and the growth regulator can be applied to large-scale seedling raising of the sweet potato virus-free seedlings.

Description

Technical Field

[0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a growth regulator for rapid propagation of sweet potato stem segments and a use method and application thereof. Background Art

[0002] The rapid propagation technology of cash crops is one of the development trends of modern agriculture. Since the culture is close to the best environmental conditions, nutrient supply and hormone regulation, and excludes competition from other organisms, it can proliferate in geometric progression. The process is divided into 6 parts: preparation of culture medium, selection and disinfection of explants, primary culture, subculture, rooting culture and seedling hardening and transplanting. In this process, primary culture is only a necessary process, while subculture is a regular and continuous process. However, after reaching a considerable number, it should be considered to transfer part of it to the rooting stage; in a sense, proliferation is only a reserve of mother plants, and rooting is the diversion of proliferation materials, which determines the production of rapid propagation products.

[0003] Sweet potato (Ipomoeabatatas Lam.), also known as sweet potato, red potato, sweet potato, red potato, red yam, etc., is a nutritious, adaptable and high-yield food crop with a sweet taste. It is rich in starch, carotene, dietary fiber, anthocyanins, vitamins and more than 10 trace elements such as potassium, iron, copper, selenium, calcium and 8 amino acids. It is known as the most balanced health food. It is also an important bioenergy and food processing raw material, and occupies an important position in agriculture and the national economy.

[0004] Sweet potatoes are root crops that use vegetative bodies for reproduction during production, so they are susceptible to virus infection, and as the cultivated varieties are cultivated for longer years, the virus density will become higher and higher. Therefore, the main method to overcome the harm of viral diseases is to use stem tips to cultivate virus-free seedlings; however, the existing method of cultivating virus-free seedlings from stem tips has a long reproduction cycle, slow sweet potato root growth, and cannot meet the technical problems of large-scale seedling cultivation needs. Summary of the invention

[0005] The present invention aims to solve the technical problem that the rapid propagation cycle of sweet potatoes in the prior art is long, the root system grows slowly and cannot meet the needs of large-scale seedling cultivation. A growth regulator for rapid propagation of sweet potato stem segments and a use method and application thereof are provided.

[0006] One of the purposes of the present invention is to provide a growth regulator for rapid propagation of sweet potato stem segments, wherein the active ingredient of the growth regulator is allantoin, and the effective concentration of the allantoin in the growth regulator is 250 mg / L.

[0007] The second object of the present invention is to provide the use of the above-mentioned growth regulator in improving the production of sweet potato virus-free seedlings, wherein the growth regulator promotes the root growth of the sweet potato virus-free seedlings.

[0008] The third object of the present invention is to provide a method for rapid propagation of sweet potato stem segments to promote root growth, the method comprising the following steps:

[0009] Add 1 / 2MS culture medium powder to the above-mentioned growth regulator with an allantoin concentration of 250 mg / L, heat to 100°C, stir evenly until dissolved, sterilize in a 120°C high pressure sterilizer for 15 minutes, and set aside; place the sweet potato stem segment in the above-mentioned sterilized 1 / 2MS culture medium for subculture to obtain sweet potato virus-free seedlings.

[0010] In a preferred embodiment of the present invention, the preparation step of the sweet potato stem segments is: when the sweet potato seedlings grown to a height of 10 cm and have 4-6 leaves after primary culture, uncontaminated sweet potato stem segments are selected and cut into 2-3 cm small segments to obtain sweet potato stem segments.

[0011] In a preferred embodiment of the present invention, the 1 / 2MS culture medium further comprises: 30 g / L sucrose, 5 g / L agar powder, and pH=5.8.

[0012] In a preferred embodiment of the present invention, the subculture environment temperature is 28° C., the environment humidity is 80%, and the light exposure time is 16 / 8 h.

[0013] Beneficial effects of the present invention: The present invention provides a growth regulator for rapid propagation of sweet potato stem segments, wherein the active ingredient of the growth regulator is allantoin, and the effective concentration of allantoin in the growth regulator is 250 mg / L. Allantoin is a hydantoin derivative and has the advantages of being safe and non-toxic. The growth regulator containing allantoin provided by the present invention, through the statistical analysis results of rooting rate and rapid propagation efficiency, can make the average growth amount of the single leaf area of ​​the virus-free sweet potato seedlings reach 0.672 cm when the allantoin concentration is 250 mg / L. 2 , root length elongation reaches 7.08cm, root fresh weight reaches 170mg, and its root system is well-developed, adventitious roots are long and numerous; it can be seen that the growth regulator containing allantoin provided by the present invention can significantly promote the rapid propagation effect of sweet potato stem segments. The growth regulator containing allantoin provided by the present invention treats sweet potato stem segments alone, and has a better rapid propagation effect than NAA alone or the growth regulator and NAA synergistically treat sweet potato stem segments, and significantly promotes the growth of the roots of the virus-free sweet potato seedlings.

[0014] The growth regulator containing allantoin provided by the present invention can be used to improve the production of sweet potato virus-free seedlings. Specifically, the growth regulator can promote the root growth of sweet potato virus-free seedlings, improve the rooting efficiency of sweet potato stem segments, significantly promote the reproduction rate of sweet potato virus-free seedlings, and can be used for large-scale seedling cultivation of sweet potato virus-free seedlings. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 The diagram is the observation result of the subculture of sweet potato for 15 days in Example 2 and Comparative Example 1: A is the control group, B is the 100 mg / L allantoin experimental group, C is the 250 mg / L allantoin experimental group, D is the 500 mg / L allantoin experimental group, E is the 750 mg / L allantoin experimental group, and F is the 1000 mg / L allantoin experimental group;

[0016] Figure 2 The diagram shows the observation results of the subculture of sweet potatoes for 15 days in Example 3 and Comparative Example 2: A is the NAA control group, and B is the NAA+alantoin experimental group. DETAILED DESCRIPTION

[0017] Those skilled in the art can refer to the content of this article and appropriately improve the process parameters. It should be particularly noted that all similar replacements and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The method and application of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the methods and applications described herein without departing from the content and scope of the present invention to implement and apply the technology of the present invention.

[0018] In order to make the purpose, technical scheme and advantages of the present invention more clear, the present invention is further described in detail below in conjunction with specific embodiments and the accompanying drawings of the specification. The test methods used in the following examples are conventional methods unless otherwise specified, and the materials, reagents, methods and instruments used are conventional materials, reagents, methods and instruments in the art unless otherwise specified, and can be obtained by those skilled in the art through commercial channels.

[0019] Example 1: Application of growth regulators in improving the production of virus-free sweet potato seedlings

[0020] S1: When the primary cultured sweet potato seedlings grow to 10 cm in height and have 4-6 leaves, uncontaminated sweet potato stem segments are selected and cut into 2-3 cm small segments to obtain sweet potato stem segments;

[0021] S2: Add 1 / 2MS culture medium powder to growth regulators with allantoin concentrations of 100, 250, 500, 750, and 1000 mg / L, respectively, heat to 100°C, stir evenly until dissolved, sterilize in a 120°C high pressure sterilizer for 15 min, and set aside; place the sweet potato stem segments in the above sterilized 1 / 2MS culture medium for subculture for 15 days, 30 bottles for each allantoin concentration group, and inoculate 4 pieces in each bottle to obtain the virus-free sweet potato seedlings treated with growth regulators of different allantoin concentrations;

[0022] The 1 / 2MS culture medium also includes: 30 g / L sucrose, 5 g / L agar powder, and pH=5.8; the environmental temperature of the subculture is 28° C., the environmental humidity is 80%, and the light exposure time is 16 / 8 h.

[0023] Note: Before inoculation, irradiate with ultraviolet light in the clean bench for about 20-30 minutes, ventilate for 10 minutes, and then perform sterile operations such as disinfection and inoculation of sweet potato stem segments; after inoculation, the material name, inoculation date and group name must be indicated on the bottle cap.

[0024] Comparative Example 1:

[0025] The difference between this comparative example and Example 1 is that the 1 / 2MS culture medium in S2 does not contain allantoin, and the rest is the same as Example 1.

[0026] Comparative Example 2: Application of growth regulators and NAA synergistically in improving the production of sweet potato virus-free seedlings

[0027] S1: When the primary cultured sweet potato seedlings grow to 10 cm in height and have 4-6 leaves, uncontaminated sweet potato stem segments are selected and cut into 2-3 cm small segments to obtain sweet potato stem segments;

[0028] S2: Add 1 / 2MS culture medium powder to a growth regulator with an allantoin concentration of 250 mg / L, heat to 100°C, stir evenly until dissolved, sterilize in a 120°C high pressure sterilizer for 15 minutes, and set aside; place the sweet potato stem segments in the above sterilized 1 / 2MS culture medium for subculture for 15 days, inoculate 30 bottles in total, and inoculate 4 pieces in each bottle to obtain the virus-free sweet potato seedlings;

[0029] The 1 / 2MS culture medium also includes: 0.5 mg / L NAA, 30 g / L sucrose, 5 g / L agar powder, and pH=5.8; the environmental temperature of the subculture is 28° C., the environmental humidity is 80%, and the light exposure time is 16 / 8 h.

[0030] Note: Before inoculation, irradiate with ultraviolet light in the clean bench for about 20-30 minutes, ventilate for 10 minutes, and then perform sterile operations such as disinfection and inoculation of sweet potato stem segments; after inoculation, the material name, inoculation date and group name must be indicated on the bottle cap.

[0031] Comparative Example 3:

[0032] The difference between this comparative example and comparative example 2 is that the 1 / 2MS culture medium in S2 does not contain allantoin, and the rest is the same as comparative example 2.

[0033] Effect experiment:

[0034] (1) The present invention uses Example 1 as the experimental group and Comparative Example 1 as the control group, and statistically analyzes the rooting rate and rapid propagation efficiency of the sweet potato stem segments in the experimental group and the control group. The rapid propagation efficiency is specifically measured from the seedling time (d), single leaf area (cm 2 A comparative analysis was conducted in four aspects: root fresh weight (mg) and root length elongation (cm).

[0035] Rapid propagation efficiency: (single seedling time of experimental group / single seedling time of control group) × 100%;

[0036] (Average fresh weight of roots in the experimental group / Average fresh weight of roots in the control group) × 100%;

[0037] (The average increase in single leaf area of ​​the experimental group / the average increase in single leaf area of ​​the control group) × 100%;

[0038] (Root length extension of experimental group / root length extension of control group × 100%).

[0039] The results are as follows Figure 1 As shown in Table 1, compared with the control group (Comparative Example 1), in the experimental group BF (Example 1) added with a growth regulator containing allantoin, the average growth of the single leaf area, the root length elongation, and the root fresh weight of the treated sweet potato virus-free seedlings were significantly improved, and the rooting rate could reach 100%. In addition, the present invention found that there were significant differences in the effects of growth regulators with different allantoin concentrations on the rapid propagation of sweet potato stem segments. In the experimental group BF (Example 1), it can be seen that low-concentration allantoin promoted the growth of the sweet potato virus-free seedlings, while high-concentration allantoin inhibited the growth of the sweet potato virus-free seedlings; when the allantoin concentration was 250 mg / L, the average growth of the single leaf area of ​​the treated sweet potato virus-free seedlings was 0.672 cm 2 , root length elongation is 7.08cm, root fresh weight is 170mg, and its root system is well-developed, the adventitious roots are long and numerous, which can significantly promote the rapid propagation of sweet potato stem segments.

[0040] Table 1

[0041]

[0042]

[0043] (2) The present invention uses the treatment group with an allantoin concentration of 250 mg / L in Example 1 as the experimental group and Comparative Examples 2-3 as the control group to conduct statistical analysis on the rooting rate and rapid propagation efficiency of sweet potato stem segments. The rapid propagation efficiency is specifically analyzed from the seedling time (d), single leaf area (cm 2 A comparative analysis was conducted in four aspects: root fresh weight (mg) and root length elongation (cm).

[0044] The results are as follows Figure 2 As shown in Table 2, compared with the control group treated with NAA alone (Comparative Example 3), the control group (Comparative Example 2) with the addition of a growth regulator containing allantoin showed an increase of 64.9% in the average growth rate of single leaf area, 54.7% in root length elongation, 156% in root fresh weight, and a rooting rate of up to 100%.

[0045] In Example 1, the experimental group with an allantoin concentration of 250 mg / L, compared with the control group (Comparative Example 2) added with a growth regulator containing allantoin, the average growth rate of single leaf area of ​​the sweet potato virus-free seedlings treated therewith increased by 41.2%, the root length elongation increased by 85.3%, and the root fresh weight increased by 32.8%.

[0046] It can be seen that compared with the treatment of sweet potato stem segments with NAA alone or the synergistic treatment of the growth regulator containing allantoin provided by the present invention, the sweet potato root system is well-developed, the adventitious roots are longer and more numerous, and the rapid propagation effect is better, which significantly promotes the growth of the roots of the sweet potato virus-free seedlings.

[0047] Table 2

[0048]

[0049] The contents not described in detail in the specification of the present invention are well-known technologies for those skilled in the art. Although the present invention has been disclosed as above with preferred embodiments, it is not intended to limit the present invention. Anyone familiar with this technology can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the protection scope of the present invention should be based on the definition of the claims.

Claims

1. A growth regulator for rapid propagation of sweet potato stem segments, characterized in that: The active ingredient of the growth regulator is allantoin, and the effective concentration of allantoin in the growth regulator is 250 mg / L.

2. The use of the growth regulator according to claim 1 in improving the production of virus-free sweet potato seedlings, characterized in that: The growth regulator promotes the root growth of the virus-free sweet potato seedlings.

3. A method for rapid propagation of sweet potato stem segments to promote root growth, characterized in that: The method comprises the following steps: Add 1 / 2MS culture medium powder to the growth regulator with an allantoin concentration of 250 mg / L as described in claim 1, heat to 100°C, stir evenly until dissolved, sterilize in a 120°C high pressure sterilizer for 15 minutes, and set aside; place the sweet potato stem segment in the sterilized 1 / 2MS culture medium for subculture to obtain sweet potato virus-free seedlings.

4. The method according to claim 3, characterized in that The preparation steps of the sweet potato stem segments are as follows: when the sweet potato seedlings grown to a height of 10 cm after primary culture and have 4 to 6 leaves, uncontaminated sweet potato stem segments are selected and cut into small segments of 2 to 3 cm to obtain sweet potato stem segments.

5. The method according to claim 3, characterized in that: The 1 / 2MS culture medium also includes: 30 g / L sucrose, 5 g / L agar powder, and pH=5.

8.

6. The method according to claim 3, characterized in that The subculture environment temperature is 28° C., the environment humidity is 80%, and the light exposure time is 16 / 8 h.

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