Method for promoting scale differentiation of longya lilium and reducing scale browning

Abstract

The invention relates to the technical field of plant tissue culture, in particular to a method for promoting scale differentiation of longya lilium and reducing scale browning. By observing scale differentiation and browning conditions under different culture medium conditions, it is found that when NAA is 0.05 mg.L <-1 >, TDZ is 0.25 mg.L <-1 >, KT is 0.05 mg.L <-1 > and AC is 6 g / L, the scale differentiation rate of the longya lilium can reach 71.66% + / -6.00 ab, the browning rate is reduced to 26.66% + / -6.00 c, the growth of the seed balls is remarkably promoted, the size of the seed balls can reach (0.52 cm + / -0.01 ab) * (0.4 cm + / -0.02a), and a support can be provided for efficient breeding of the longya lilium seed balls.

Description

Technical Field

[0001] The invention relates to the technical field of plant tissue culture, and in particular to a method for promoting differentiation of Lilium aralia scales and reducing browning of scales. Background Art

[0002] Dragon Tooth Lily (Lilium brownii var. viridulum) is a perennial herbaceous plant of the genus Lilium in the family Liliaceae. It is a variant of the wild lily (Lilium brownii) and is named for its enlarged scales that resemble dragon teeth. It is mainly produced in Shaoyang, Longhui in Hunan and Wanzai, Taihe in Jiangxi. It is one of the first batch of medicinal and edible foods announced by the Ministry of Health and is deeply favored by consumers in Southeast Asian countries and regions. In production, Dragon Tooth Lily mainly reproduces asexually by scale cuttings and bulb division, but this method will lead to the degeneration of its species, and the yield and quality will be significantly reduced. In addition, Dragon Tooth Lily also has the problems of low differentiation coefficient and long breeding cycle during the breeding process. Therefore, high-quality and efficient breeding of bulbs is the only way to solve the development of the Dragon Tooth Lily industry.

[0003] Plant tissue culture technology is one of the important research technologies and means in life sciences. Tissue culture technology can accelerate the reproduction speed of lilies, shorten the growth cycle of lily bulbs, and make up for the shortcomings of bulb breeding. Therefore, it is widely used in the commercialization and industrialization of lilies. In recent years, there have been many research reports on lily tissue culture at home and abroad. Lily tissue culture often uses scales, stem segments, etc. as explants, adds 6-BA, NAA, 2,4-D and other hormones, and promotes the induction, proliferation and expansion of lily bulbs in a single or combined form. For example, the Chinese invention patent with publication number CN 107396835 A discloses a method for rapid propagation of edible lilies. The invention greatly improves the adventitious bud induction effect of famous and excellent varieties of lily explants by determining the culture medium and culture conditions at each stage of the proliferation of famous and excellent varieties of lily explants, and at the same time, the adventitious bud differentiation rate is high and the differentiation amount is large. However, this method can only promote the differentiation of lily scales, and there is still the problem of browning of scales during the breeding and cultivation of lilies. In addition, the lily genome is huge, with a high degree of heterozygosity, different varieties have different ploidy, and there are huge differences between different hybrid lines. The hormone treatment combinations required for different varieties of lily are different. Therefore, this method cannot be widely used. Summary of the invention

[0004] In view of the above-mentioned deficiencies in the prior art, the object of the present invention is to provide a method for promoting differentiation of Lilium arborvitae scales and reducing browning of scales.

[0005] In order to achieve the above-mentioned object of the invention, the technical solution adopted by the present invention is:

[0006] Provided is a method for promoting differentiation of Lilium argentea scales and reducing browning of scales, which is achieved by setting a culture medium and culture conditions; the culture medium and culture conditions are: MS+NAA 0.05 mg·L -1 +TDZ 0.25mg·L -1 +KT 0.05mg·L -1 +AC 6g·L -1 , pH 6.0.

[0007] Furthermore, the method specifically includes the following steps:

[0008] Step 1: Select the white scales without spots in the middle of the bulb of Lilium longituba and clean them under running water;

[0009] Step 2: Prepare culture medium: MS + NAA 0.05 mg·L -1 +TDZ 0.25mg·L -1 +KT 0.05mg·L -1 +AC6g·L -1 , pH 6.0, the culture medium was sterilized at high temperature before use;

[0010] Step 3: Place the scales in a sterilized empty bottle and place them on a clean bench for UV sterilization for 15 minutes, treat with 75% alcohol for 30 seconds, then treat with 10% NaClO for 15 minutes, and rinse with sterile water 3 times, 3 minutes each time;

[0011] Step 4, remove the transparent part around the scale, cut the scale into two parts, and inoculate them into the culture medium with the concave surface facing upwards;

[0012] Step 5: After 20 days, observe the differentiation and browning of the lily scales, calculate the differentiation rate, and determine the size of the seed ball.

[0013] Furthermore, activated carbon AC needs to be added to the culture medium at the end.

[0014] The beneficial effects of the present invention are:

[0015] The present invention proposes a method for promoting the differentiation of Lilium argentea scales and reducing the browning of scales, which can effectively improve the differentiation rate of Lilium argentea scales and reduce the browning of Lilium argentea scales, shorten the growth time of seed balls and significantly promote the growth of seed balls. -1 +TDZ 0.25mg·L -1 +KT 0.05mg·L -1 The optimal medium for inducing the bulblets of Lilium argenteae was found, and the differentiation rate of scales could reach 81.67%±3.33a. -1The differentiation rate after AC was 71.66%±6.00ab, which was not significantly different from the control, but the browning rate was significantly reduced to 26.66%±6.00c, and the ball size could reach (0.52cm±0.01ab)×(0.4cm±0.02a) (length×width), which was significantly higher than other groups, providing support for the efficient breeding of Lilium bulbs. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 The growth of 20-day scales of Lilium truncatum under different differentiation medium conditions in the embodiment (Table 1); the numbers in the figure represent the treatment numbers;

[0017] Figure 2 The growth conditions of Lilium argenteae under different anti-browning culture media (Table 2) in the example are shown; the numbers in the figure represent different treatment concentrations of activated carbon. DETAILED DESCRIPTION

[0018] The specific implementation modes of the present invention are described below so that those skilled in the art can understand the present invention. However, it should be clear that the present invention is not limited to the scope of the specific implementation modes. For those of ordinary skill in the art, as long as various changes are within the spirit and scope of the present invention as defined and determined by the attached claims, these changes are obvious, and all inventions and creations utilizing the concept of the present invention are protected.

[0019] Test example

[0020] In this experiment, we first conducted a differentiation test on Lilium argentea scales and screened out the most suitable differentiation medium and culture conditions: MS + NAA 0.05 mg·L -1 +TDZ 0.25mg·L -1 +KT 0.05mg·L -1 , pH 6.0, and then a browning test was performed, in which different concentrations of activated carbon were added to the optimal medium for the differentiation test.

[0021] The specific steps of the differentiation assay are:

[0022] Step 1: Select the white scales without spots in the middle of the bulb of Lilium longituba and clean them under running water;

[0023] Step 2, preparing the scale differentiation culture medium treated with different hormones, and using the culture medium after high temperature sterilization;

[0024] Step 3: Place the scales in a sterilized empty bottle and place them on a clean bench for UV sterilization for 15 minutes, treat with 75% alcohol for 30 seconds, then treat with 10% NaClO for 15 minutes, and rinse with sterile water 3 times, 3 minutes each time;

[0025] Step 4, remove the transparent part around the scale, cut the scale into two parts, and inoculate them into different culture media with the concave surface facing upwards;

[0026] Step 5: Observe the differentiation of lily scales after 20 days and calculate the differentiation rate.

[0027] Table 1

[0028]

[0029] The specific steps of the browning test are:

[0030] Step 1: Select the white scales without spots in the middle of the bulb of Lilium longituba and clean them under running water;

[0031] Step 2: In differentiation medium MS + NAA 0.05 mg L -1 +TDZ 0.25mg·L -1 +KT 0.05mg·L -1 Different concentrations of activated carbon AC were added to the culture medium (Table 2), where AC was added to the culture medium at the end and the culture medium was sterilized at high temperature before use;

[0032] Step 3: Place the scales in a sterilized empty bottle and place them on a clean bench for UV sterilization for 15 minutes, treat with 75% alcohol for 30 seconds, then treat with 10% NaClO for 15 minutes, and rinse with sterile water 3 times, 3 minutes each time;

[0033] Step 4, remove the transparent part around the scale, cut the scale into two parts, and inoculate them into the culture medium with the concave surface facing upwards;

[0034] Step 5: After 20 days, observe the differentiation and browning of the lily scales, calculate the differentiation rate, and determine the size of the seed ball.

[0035] Table 2

[0036]

[0037] In summary, the present invention observed the differentiation and browning of scales under different culture medium conditions and found that when NAA was 0.05 mg·L -1 , TDZ is 0.25 mg·L -1 , KT is 0.05 mg·L -1 When AC is 6g / L, the differentiation rate of Lilium argentea scales can reach 71.66%±6.00ab, the browning rate is reduced to 26.66%±6.00c, and the growth of seed balls is significantly promoted. The size of the seed balls can reach (0.52cm±0.01ab)×(0.4cm±0.02a)(length×width), which can provide support for the efficient breeding of Lilium argentea bulbs.

[0038] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the present invention can be implemented in other specific forms without departing from the spirit or essential features of the present invention. Therefore, the embodiments should be considered exemplary and non-restrictive in all respects, and the scope of the present invention is defined by the appended claims rather than the above description, and it is intended that all changes falling within the meaning and scope of the equivalent elements of the claims be included in the present invention.

[0039] In addition, it should be understood that although the present specification is described according to implementation modes, not every implementation mode contains only one independent technical solution. This description of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment may also be appropriately combined to form other implementation modes that can be understood by those skilled in the art.

Claims

1. A method for promoting differentiation of Lilium aralia scales and reducing browning of scales, characterized in that: This is achieved by setting the culture medium and culture conditions; the culture medium and culture conditions are: MS+NAA 0.05mg·L -1 +TDZ 0.25mg·L -1 +KT0.05mg·L -1 +AC 6g·L -1 , pH 6.

0.

2. The method for promoting differentiation of Lilium aralia scales and reducing browning of scales according to claim 1, characterized in that: The specific steps include: Step 1: Select the white scales without spots in the middle of the bulb of Lilium longituba and clean them under running water; Step 2: Prepare culture medium: MS + NAA 0.05 mg·L -1 +TDZ 0.25mg·L -1 +KT 0.05mg·L -1 +AC 6g·L -1 , pH 6.0, the culture medium was sterilized at high temperature before use; Step 3: Place the scales in a sterilized empty bottle and place them on a clean bench for UV sterilization for 15 minutes, treat with 75% alcohol for 30 seconds, then treat with 10% NaClO for 15 minutes, and rinse with sterile water 3 times, 3 minutes each time; Step 4: Remove the transparent part around the scales, cut the scales into two parts, and inoculate them into the culture medium with the concave side facing upwards.

3. The method for promoting differentiation of Lilium aralia scales and reducing browning of scales according to claim 1 or 2, characterized in that: Activated carbon AC needs to be added to the culture medium last.

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