CN114667929A - Method for changing leaf color of tissue culture seedlings of ribgrass red leaf by utilizing illumination - Google Patents
Method for changing leaf color of tissue culture seedlings of ribgrass red leaf by utilizing illumination 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
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
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- A01G7/045—Electric or magnetic or acoustic treatment of plants for promoting growth with electric lighting
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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
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- A—HUMAN NECESSITIES
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- 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
- A01H4/002—Culture media for tissue culture
Abstract
The invention discloses a method for changing the leaf color of a tissue culture seedling of ribwort redleaf by utilizing illumination. The method is characterized in that the rough costaria verrucosa bud is cultured under the condition that the illumination intensity is 4000lx or the light quality condition is red light and blue light is 3:7 in the proliferation, differentiation and rooting culture stage, and the illumination intensity and the light quality condition in the proliferation, differentiation and rooting culture stage of the rough costaria rucosa bud are controlled, so that the red and bright degree of the leaf color of the rough costaria rucosa can be effectively kept, and the efficient, rapid and large-scale propagation of high-quality rough costaria rucosa seedlings is facilitated.
Description
Technical Field
The invention belongs to the field of plant seedling production, and particularly relates to a method for changing the leaf color of a tissue culture seedling of ribwort rupestris by utilizing illumination.
Background
Roughhaired costwort (Aglaonema spp.) is a perennial herbaceous foliage plant of the genus Roughhaired costwort (Aglaonema) of the family Araceae (Araceae). The red-leaf burley grass is one of the popular color-leaf varieties in China in recent years, is widely used indoors as a household small potted plant, is relatively shade-resistant, has long indoor arrangement and appreciation period, and can purify air. And because the leaves have larger red patches, the middle ribs are large and the color is bright, the defect of monotonous color indoors and outdoors in light flower seasons can be overcome, the red-leaf burley grass is deeply loved by consumers, and the planting and market prospect of the red-leaf burley grass is good. Along with the increasing year by year of the seedling yield of the tissue culture of the rudbeckia rossica, the problems that the leaf color is faded and turns green, the red and bright degree of the leaf color is not strong and the like easily occur to partial seedlings and finished potted flowers of the rudbeckia rossica variety, and the large-scale production and popularization of the rudbeckia rossica variety are greatly not facilitated. The red-leaf burley grass propagation mode mainly comprises the branch propagation, the cutting propagation, the tissue culture rapid propagation and the like, and the two traditional propagation modes of the branch propagation and the cutting propagation are easily infected by germs due to improper wound treatment, and the propagation rate is correspondingly lower; the tissue culture and rapid propagation mode is increasingly the main mode for supplying the seedlings of the Roxburgh goosegrass, but the leaf color and the red brilliance of the Roxburgh goosegrass seedlings propagated by the tissue culture and rapid propagation mode always have stability to a great extent, and the tissue culture and rapid propagation mode also becomes the bottleneck for limiting the propagation of the Roxburgh goosegrass seedlings and promoting the industrial development. Therefore, the research and discussion of the factors influencing the leaf color of the red-leaf burley grass in the tissue culture process have very important significance for maintaining the red brilliance of the leaf color and improving the tissue culture production efficiency.
The invention aims to explore factors influencing the leaf color in the process of tissue culture of the ribgrass redleaf from the aspect of culture conditions (illumination) of the ribgrass redleaf. The feasibility of keeping the color brightness of the leaves in the tissue culture link is discussed, and technical references are provided for further stabilizing the efficient tissue culture technology of the red-leaf type coarse ribbed grass and regulating the leaf color.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provides a method for changing the leaf color of a tissue culture seedling of ribgrass carpesium rubrum by utilizing illumination.
The technical scheme adopted by the invention is as follows:
a method for changing the leaf color of a tissue culture seedling of ribgrass carpesium rubrum by utilizing illumination comprises the following steps: culturing the thick burley bush buds under certain illumination intensity or light quality in the proliferation differentiation and rooting culture stages.
Preferably, the method for changing the leaf color of the tissue culture seedlings of the ribwort herb red leaves by utilizing illumination comprises the following steps: culturing the rough cord grass clump buds under the condition that the illumination intensity is 4000lx or the light quality condition is red light and blue light is 3:7 in the proliferation, differentiation and rooting culture stages.
Preferably, the method for changing the leaf color of the tissue culture seedlings of the ribwort redleaf by using illumination comprises the following steps: culturing the rough cord grass clump buds under the conditions of illumination intensity of 4000lx, illumination time of 10h/d and temperature of 25 +/-1 ℃ in the proliferation differentiation and rooting culture stages.
Preferably, the method for changing the leaf color of the tissue culture seedlings of the ribwort redleaf by using illumination comprises the following steps: culturing the rough cord grass clump buds under the condition that the light quality condition is red light, blue light is 3:7, the illumination time is 10h/d, and the temperature is 25 +/-1 ℃ in the proliferation differentiation and rooting culture stages.
Preferably, the multiple shoots are induced from the stem of the rough burclover as an explant.
Preferably, the formula of the culture medium for proliferation and differentiation culture is as follows: MS + KT 0.2mg/L +6-BA 3.0mg/L + IBA 0.02mg/L + sucrose 30g/L + agar 5.5g/L, pH 6.0.
Preferably, the formula of the rooting culture medium is as follows: MS + NAA0.5mg/L + AC 0.2g/L + sucrose 30g/L + agar 5.5g/L, pH 6.0.
The innovation of the invention is to disclose a key technology of a method for changing the leaf color of the tissue culture seedlings of the ribgrass carpesium rubrum by utilizing illumination. The innovation of the invention is that: the rough-costed grass clump buds are cultured under the condition that the illumination intensity is 4000lx or the light quality condition is red light and blue light is 3:7 in the proliferation, differentiation and rooting culture stages, and the red brilliance of the leaf color of the rough-costed grass clump buds can be effectively maintained by controlling the illumination intensity and the light quality condition in the proliferation, differentiation and rooting culture stages, so that the high-quality rough-costed grass seedlings can be efficiently and quickly propagated in a large scale.
Drawings
FIG. 1 is a graph of the effect of different illumination intensities on leaf color. a: the illumination intensity is as follows: 3000 lx; b: the illumination intensity is as follows: 4000 lx.
FIG. 2 is a graph of the effect of different light quality conditions on leaf color. a: white light; b: red light (100% R); c: red-blue 7:3 (70% R: 30% B); d: red-blue-5: 5 (50% R: 50% B); e: red-blue 3:7 (30% R: 70% B); f: blue light (100% B); g: purple light.
Detailed Description
The following examples are further illustrative of the present invention and are not intended to be limiting thereof.
The test material 'Guangzhou red burley grass' in the following examples is a self-bred variety of Guangzhou flower research center, and the tissue culture material is induced by taking a burley grass stem section as an explant.
Example 1: illumination intensity processing
1. Method of producing a composite material
Treatment C1: 3000lx, treatment C2: 4000lx, the illumination time is 10h/d, and the temperature is 25 +/-1 ℃.
Taking a 'Guanghua burclover' stem section as an explant, treating the sterilized explant by adopting a cluster bud induced proliferation mode for proliferation and propagation, respectively culturing sterile bottle seedlings in proliferation and differentiation and rooting culture stages under the conditions of C1 and C2, wherein the illumination time is 10h/d, the temperature is 25 +/-1 ℃, the proliferation culture time is 50d, the number of times of repetition is 3, and the culture medium formula of the proliferation and differentiation culture is as follows: MS + KT 0.2mg/L +6-BA 3.0mg/L + IBA 0.02mg/L + sucrose 30g/L + agar 5.5g/L, pH 6.0; measuring the height, the leaf length and the leaf width of each single seedling obtained by proliferation and differentiation, and measuring 30 seedlings in each treatment; continuing to perform root transformation culture for 40 days, wherein the formula of the rooting culture medium is as follows: MS + NAA0.5mg/L + AC 0.2g/L + sucrose 30g/L + agar 5.5g/L, pH 6.0; observing the leaf color, counting the area ratio of the red patch leaves, transplanting the red patch leaves to a greenhouse, and counting the elimination rate of the grown seedlings after 120 days.
2. Data processing
The elimination rate (%) × 100% (number of abnormal seedlings such as non-red leaf color, abnormal shape, and mutation/total planted plants);
the seedling rate (%) is 1-elimination rate (%).
Carrying out statistical arrangement on experimental data by using Excel 2010 office software; analysis of variance and multiple comparisons (Duncan) were performed using Statistical Analysis software SAS 9.2(Statistical Analysis System).
3. Results
The results are shown in Table 1 and FIG. 1. The proliferation and differentiation seedlings cultured by two illumination intensities are characterized in that the illumination intensity of 3000lx (C1) is 3000lx, the plant height is 3.42cm, the plant height is obviously higher than that of 4000lx (C2) by 2.95cm, but the leaf width is 1.89cm, the leaf length is 2.49cm, the leaf width/leaf length is 0.77, and the leaf width, leaf length and leaf width/leaf length are all less than that of 4000 lx; the seedlings are proliferated and differentiated under 3000lx illumination intensity, the plants are thin and high, the leaf area is small, and the proportion of leaves is long; the proliferation and differentiation seedlings with the illumination intensity of 4000lx have shorter and stronger plants, large leaf area and round leaf proportion. In the aspect of leaf color, 3000lx illumination intensity, the leaf of the rooted seedling has light red color and small red patch percentage (figure 1a), and in the aspect of 4000lx illumination intensity, the red patch percentage is large and the leaf color is red (figure 1 b). After transplanting and planting, the elimination rate of the rooting seedlings with the illumination intensity of 4000lx is 11.11 percent and is obviously lower than that of the rooting seedlings with the illumination intensity of 3000 lx. The result shows that the multiplication and differentiation seedlings with the illumination intensity of 4000lx are shorter and stronger, the color of the leaves of the rooting seedlings is better, the planting elimination rate is lower, and the method is more suitable for the tissue culture illumination culture of the red-leaf burley grass.
TABLE 1 Effect of different illumination intensities on the growth and leaf color of tissue culture seedlings
Note: the difference of different lower case letters in the same column is obvious, and P is less than 0.05; plaque grade (red plaque proportion): +: less than 20 percent; ++: 20 to 50 percent; +++: 50% -80%; ++++: more than 80 percent.
Example 2: light quality test
1. Method of producing a composite material
Seven treatments are set up: d1: white light (LED lamps); d2: red light (100% R); d3: red-blue 7:3 (70% R: 30% B); d4: red-blue 5:5 (50% R: 50% B); d5: red-blue 3:7 (30% R: 70% B); d6: blue light (100% B); d7: purple light; the illumination time is 10h/d, and the temperature is 25 +/-1 ℃.
Taking a 'Guanghua burclover' stem section as an explant, treating the sterilized explant by a cluster bud induced proliferation mode for proliferation and propagation, respectively culturing sterile bottle seedlings in proliferation differentiation and rooting culture stages under the conditions of D1-D7, wherein the illumination time is 10h/D, the temperature is 25 +/-1 ℃, the proliferation culture time is 50D, 10 bottles/treatment are carried out, the repetition times are set to be 3 times, the formula of a culture medium for proliferation and differentiation culture is MS + KT 0.2mg/L +6-BA 3.0mg/L + IBA 0.02mg/L + sucrose 30g/L + agar 5.5g/L, and the pH is 6.0; then continuing to carry out root transformation culture for 40 days, wherein the formula of a rooting culture medium is as follows: MS + NAA0.5mg/L + AC 0.2g/L + sucrose 30g/L + agar 5.5g/L, pH 6.0. Observing the leaf color of the tissue culture seedling, taking the 1 st to 2 nd flat leaves counted from top to bottom, and extracting and measuring the content of chlorophyll and anthocyanin.
Chlorophyll extraction and content determination: reference is made to the related methods of the literature "Liheng. plant physiology and biochemistry experiment principle and technology [ M ]. Beijing: advanced education Press, 2000: 130-.
Anthocyanin extraction and determination: reference is made to the literature "Wangyanwana Siji begonia leaf transcriptome sequencing analysis and low temperature induced anthocyanin synthesis mechanism [ D ]. Henan university of agriculture, 2018." and "Liu Xiao Dong, in the methods related to the extraction of anthocyanin from leaves of Plectranthus niveus (purple leaf) and its stability [ J ]. proceedings of northeast university of forestry, 2011,39(02):38-39+81 ].
2. Data processing
The same as in example 1.
3. Results
The results of the effect of 7 different light quality treatments on the content of photosynthetic pigments in leaves can be seen (table 2), the content of chlorophyll a + b irradiated by pure red light (D2) is the highest and is 0.058mg/g, and the difference with the white light (D1) and purple light (D7) irradiation groups is not significant, but is significantly higher than that of other light treatment groups; the relative content of anthocyanin is the lowest, and is 0.056Units/g, and the anthocyanin is not different from the white light (D1) and purple light (D7) treated group, but is obviously lower than other light-treated groups. Irradiating red light, wherein the total content of chlorophyll a and b in blue light of 3:7(D5) is the lowest and is 0.037mg/g, and the red light is not significantly different from that in a pure blue light (D6) irradiation group and is obviously lower than that in other light quality treatment groups; the relative anthocyanin content was highest, 0.399Units/g, and all significantly higher than the other treatment groups. The results show that pure red light is not beneficial to the accumulation of anthocyanin in the leaves of the ruddy-leaf ruddy grass, the leaf color is the worst, and the leaves are green-white (figure 2 b); blue light promotes the accumulation of anthocyanin under the condition of a certain light quality proportion, wherein the light quality of red light, namely 3:7, is the highest relative content of anthocyanin, the leaf color is red and bright (figure 2e), and pure blue light, red light, namely 7:3, red light, namely 5:5 are adopted.
TABLE 2 Effect of different light quality conditions on photosynthetic pigment content in leaves
Note: the difference is marked by different lower case letters in the same column, and P is less than 0.05.
Example 3
A method for changing the leaf color of a tissue culture seedling of ribgrass carpesium rubrum by utilizing illumination comprises the following steps: using the stem segment of the disinfected Crabapple cinquefoil herb as an explant to induce cluster buds, wherein the cluster bud induction culture medium comprises: 1/2MS +6-BA 2.0mg/L + sucrose 30g/L + agar 6.0g/L, pH 6.0; inoculating the cluster buds into a multiplication culture medium to culture under the conditions of illumination intensity of 4000lx, illumination time of 10h/d and temperature of 25 +/-1 ℃, wherein the formula of the multiplication culture medium is as follows: MS + KT 0.2mg/L +6-BA 3.0mg/L + IBA 0.02mg/L + sucrose 30g/L + agar 5.5g/L, pH is 6.0; placing the single seedlings obtained by proliferation and differentiation into a rooting culture medium, wherein the formula of the rooting culture medium is as follows: MS + NAA0.5mg/L + AC 0.2g/L + sucrose 30g/L + agar 5.5g/L, pH 6.0: culturing under the conditions of illumination intensity of 4000lx, illumination time of 10h/d and temperature of 25 +/-1 ℃; transplanting the rooted seedlings.
Example 4
A method for changing the leaf color of a tissue culture seedling of ribgrass carpesium rubrum by utilizing illumination comprises the following steps: using the stem segment of the disinfected Crabapple cinquefoil herb as an explant to induce cluster buds, wherein the cluster bud induction culture medium comprises: 1/2MS +6-BA 2.0mg/L + sucrose 30g/L + agar 6.0g/L, pH 6.0; inoculating the cluster buds into a multiplication culture medium to be cultured under the conditions of red light, blue light, 3:7 (30% R, 70% B), illumination time of 10h/d and temperature of 25 +/-1 ℃, wherein the formula of the multiplication culture medium is as follows: MS + KT 0.2mg/L +6-BA 3.0mg/L + IBA 0.02mg/L + sucrose 30g/L + agar 5.5g/L, pH 6.0; placing the single plant seedlings obtained by proliferation and differentiation into a rooting culture medium, wherein the formula of the rooting culture medium is as follows: MS + NAA0.5mg/L + AC 0.2g/L + sucrose 30g/L + agar 5.5g/L, pH 6.0, culturing under the condition of red light, blue light 3:7 (30% R: 70% B), illumination time 10h/d, and temperature 25 + -1 deg.C; transplanting the rooted seedlings.
The above are only preferred embodiments of the present invention, and it should be noted that the above preferred embodiments should not be considered as limiting the present invention, and the protection scope of the present invention should be subject to the scope defined by the claims. It will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the spirit and scope of the invention, and these modifications and adaptations should be considered within the scope of the invention.
Claims (7)
1. A method for changing the leaf color of a tissue culture seedling of ribgrass bicolor by utilizing illumination is characterized by comprising the following steps: culturing the thick burley bush buds under certain illumination intensity or light quality in the proliferation differentiation and rooting culture stages.
2. The method for changing the leaf color of the tissue culture seedlings of the ribgrass bicolor by using illumination according to claim 1, which comprises the following steps: culturing the rough cord grass clump buds under the condition that the illumination intensity is 4000lx or the light quality condition is red light and blue light is 3:7 in the proliferation, differentiation and rooting culture stages.
3. The method for changing the leaf color of the tissue culture seedlings of the ribgrass carpesium rubrum by using illumination as claimed in claim 2, comprising the steps of: culturing the rough cord grass clump buds under the conditions of illumination intensity of 4000lx, illumination time of 10h/d and temperature of 25 +/-1 ℃ in the proliferation differentiation and rooting culture stages.
4. The method for changing the leaf color of the tissue culture seedlings of the ribgrass carpesium rubrum by using illumination as claimed in claim 2, comprising the steps of: culturing the rough cord grass clump buds under the condition that the light quality condition is red light, blue light is 3:7, the illumination time is 10h/d, and the temperature is 25 +/-1 ℃ in the proliferation differentiation and rooting culture stages.
5. The method for changing the leaf color of a ribbed red-leaf tissue culture seedling by illumination according to any one of claims 1-4, wherein the multiple shoots are induced by using ribbed grass stem as an explant.
6. The method for changing the leaf color of a ribwort tissue culture seedling by using illumination according to any one of claims 1 to 4, wherein the formula of the culture medium for proliferation and differentiation culture is as follows: MS + KT 0.2mg/L +6-BA 3.0mg/L + IBA 0.02mg/L + sucrose 30g/L + agar 5.5g/L, pH 6.0.
7. The method for changing the leaf color of the ribbed red-leaf ribbed grass tissue culture seedling by illumination according to any one of claims 1-4, characterized in that the formula of the culture medium for rooting culture is as follows: MS + NAA0.5mg/L + AC 0.2g/L + sucrose 30g/L + agar 5.5g/L, pH 6.0.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080209598A1 (en) * | 2004-06-15 | 2008-08-28 | Syngenta Participations Ag | Plant Cultivation Method |
| CN107711513A (en) * | 2017-11-24 | 2018-02-23 | 广东省农业科学院环境园艺研究所 | A kind of thick rib grass quick breeding method for tissue culture |
| CN109220804A (en) * | 2018-11-05 | 2019-01-18 | 吴子平 | A kind of high efficiency quick breeding method of the thick rib grass of coloured silk leaf |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080209598A1 (en) * | 2004-06-15 | 2008-08-28 | Syngenta Participations Ag | Plant Cultivation Method |
| CN107711513A (en) * | 2017-11-24 | 2018-02-23 | 广东省农业科学院环境园艺研究所 | A kind of thick rib grass quick breeding method for tissue culture |
| CN109220804A (en) * | 2018-11-05 | 2019-01-18 | 吴子平 | A kind of high efficiency quick breeding method of the thick rib grass of coloured silk leaf |
Non-Patent Citations (2)
| Title |
|---|
| 周佐葡: "彩叶粗肋草离体培养及影响叶色因子研究", 《中国优秀硕士学位论文全文数据库-农业科技辑》 * |
| 顾梦云等: "红叶粗肋草组织培养叶色影响因素研究（网络首发）", 《广东农业科学》 * |
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