CN108849498B - Method for promoting accumulation of flavonoid substances in leaf tissues of ornamental crabapple - Google Patents
Method for promoting accumulation of flavonoid substances in leaf tissues of ornamental crabapple Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 44
- 229930003935 flavonoid Natural products 0.000 title claims abstract description 37
- 150000002215 flavonoids Chemical class 0.000 title claims abstract description 37
- 235000017173 flavonoids Nutrition 0.000 title claims abstract description 37
- 239000000126 substance Substances 0.000 title claims abstract description 34
- 238000009825 accumulation Methods 0.000 title claims abstract description 26
- 230000001737 promoting effect Effects 0.000 title claims abstract description 22
- 235000005087 Malus prunifolia Nutrition 0.000 title claims description 30
- 244000070406 Malus silvestris Species 0.000 title 1
- YJPIGAIKUZMOQA-UHFFFAOYSA-N Melatonin Natural products COC1=CC=C2N(C(C)=O)C=C(CCN)C2=C1 YJPIGAIKUZMOQA-UHFFFAOYSA-N 0.000 claims abstract description 55
- DRLFMBDRBRZALE-UHFFFAOYSA-N melatonin Chemical compound COC1=CC=C2NC=C(CCNC(C)=O)C2=C1 DRLFMBDRBRZALE-UHFFFAOYSA-N 0.000 claims abstract description 55
- 229960003987 melatonin Drugs 0.000 claims abstract description 55
- 229930002877 anthocyanin Natural products 0.000 claims abstract description 23
- 235000010208 anthocyanin Nutrition 0.000 claims abstract description 23
- 239000004410 anthocyanin Substances 0.000 claims abstract description 23
- 150000004636 anthocyanins Chemical class 0.000 claims abstract description 23
- 241001481296 Malus spectabilis Species 0.000 claims abstract description 21
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- 238000013518 transcription Methods 0.000 claims abstract description 14
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- 241000218993 Begonia Species 0.000 claims description 11
- 238000005286 illumination Methods 0.000 claims description 11
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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
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
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Abstract
The invention discloses a method for promoting accumulation of flavonoid substances in leaf tissues of normal-color purple-leaf ornamental Chinese flowering crabapple, which comprises the following steps: the method comprises the steps of designing melatonin treatment ornamental malus spectabilis tissue culture seedlings with different concentration gradients by taking ornamental malus spectabilis leaf varieties as materials, screening 200 mu g/l of melatonin treatment leaves according to phenotypes, wherein the leaf colors are very obvious, the color of the leaves with high concentration or low concentration is not obviously changed, and subsequently detecting the content of flavonoid substances in the 200 mu g/l of melatonin treatment ornamental malus spectabilis leaves, the transcription level of key genes in a flavonoid metabolic pathway and the transcription level of genes related to the melatonin pathway, wherein results show that the anthocyanin content is highest in the 200 mu g/l of melatonin treatment ornamental malus spectabilis. The invention provides a method for improving the phenomenon of the ancestral of the ornamental Chinese flowering crabapple due to the increase of the subculture frequency, and further provides a good theoretical basis for the genetic improvement of the future flowering plants.
Description
Technical Field
The invention relates to an application method in ornamental malus spectabilis tissue culture, in particular to a method for promoting accumulation of flavonoid substances in leaf tissues of ornamental malus spectabilis.
Background
Color is an important characteristic of many plant exteriors. However, different color modules can be observed in ornamental begonia leaf tissue. The special leaf color type of the ornamental crabapple also provides a good research material for researching the color forming mechanism of the color leaf plant, which changes the current situation that the ornamental crabapple resource is in stock and sporadic cultivation for a long time in China, and expands the research content and application field of the fruit tree plant as an ornamental, medicinal and greening tree species. At present, in the breeding process of ornamental Chinese flowering crabapple seedlings, tissue culture is an important means for producing virus-free seedlings, and in the tissue culture process, the phenomena of variety degradation, leaf fading and the like are easy to occur. However, in the tissue culture process, the research on the rapid promotion of the content of flavonoid substances in the leaf tissue and further the promotion of the color of the ornamental crabapple leaves is rarely reported.
Disclosure of Invention
Based on the problems in the prior art, the invention aims to provide a method for promoting the accumulation of flavonoid substances in the leaf tissue of ornamental malus spectabilis, which can promote the accumulation of the flavonoid substances in the leaf tissue of the ornamental malus spectabilis and quickly promote the color and luster of the leaf of the ornamental malus spectabilis.
The purpose of the invention is realized by the following technical scheme:
the embodiment of the invention provides a method for promoting accumulation of flavonoid substances in ornamental begonia leaf tissues, which comprises the following steps:
taking a tissue culture seedling of an ornamental crabapple leaf variety as a material, respectively adding the tissue culture seedling of the variety into melatonin culture media with different concentrations for propagation, and then culturing for 2 weeks under normal illumination;
selecting leaf samples with obvious phenotype and good growth vigor, extracting RNA from the leaf samples, inverting the RNA into cDNA to perform qRT-PCR experiment, detecting the transcription level of key structure genes in an anthocyanin metabolic pathway, detecting the content of flavonoid substances, and determining the optimal concentration of a melatonin culture medium which is processed by the optimal anthocyanin accumulation amount, the expression amount of the anthocyanin metabolic genes and the content of the flavonoid substances correspondingly as the optimal concentration of the melatonin culture medium for promoting the accumulation of the flavonoid substances in the leaf tissues of the ornamental crabapple;
and performing propagation culture on the tissue culture seedling of the ornamental crabapple leaf variety by using the determined melatonin culture medium with the optimal concentration.
According to the technical scheme provided by the invention, the method for promoting the accumulation of flavonoid substances in the leaf tissue of ornamental crabapple has the following beneficial effects:
aiming at the aspect of the leaf color of the ornamental malus spectabilis, a method for quickly promoting the leaf color of the ornamental malus spectabilis is constructed, the method can quickly and efficiently promote the content of flavonoid substances in leaf tissues of the ornamental malus spectabilis, can improve the transcription level of key genes in an anthocyanin metabolic pathway, can produce a large number of tissue culture seedlings with excellent leaf tissue color in a short time, and overcomes the defects that the number of subculture of the ornamental malus spectabilis is increased and the leaf color is degraded.
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In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the description of the embodiments are briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on the drawings without creative efforts.
Fig. 1 is a diagram illustrating the content of flavonoid in the tissue of the 'royal' leaves after treatment with melatonin of various concentrations according to an embodiment of the present invention;
fig. 2 is a schematic representation of transcription levels of anthocyanin metabolism key structural genes in 'royal' leaf tissue after treatment with melatonin of different concentrations, provided by an embodiment of the present invention;
fig. 3 is a schematic representation of the transcription levels of other structural genes of anthocyanin metabolism in 'royal' leaf tissue after treatment with melatonin of various concentrations provided by an embodiment of the present invention;
fig. 4 is a schematic diagram of the expression levels of anthocyanin metabolism transcription factors in 'royal' leaf tissue after treatment with different concentrations of melatonin provided by the embodiment of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention are clearly and completely described below with reference to the specific contents of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments of the present invention without making any creative effort, shall fall within the protection scope of the present invention. Details which are not described in detail in the embodiments of the invention belong to the prior art which is known to the person skilled in the art.
The embodiment of the invention provides a method for promoting accumulation of flavonoid substances in ornamental begonia leaf tissues, which can quickly promote accumulation of the flavonoid substances in the ornamental begonia leaf tissues, thereby improving ornamental and medicinal values of flowering plants, and comprises the following steps:
taking a tissue culture seedling of an ornamental crabapple leaf variety as a material, respectively adding the tissue culture seedling of the variety into melatonin culture media with different concentrations for propagation, and then culturing for 2 weeks under normal illumination;
selecting leaf samples with obvious phenotype and good growth vigor, extracting RNA from the leaf samples, inverting the RNA into cDNA to perform qRT-PCR experiment, detecting the transcription level of key structure genes in an anthocyanin metabolic pathway, detecting the content of flavonoid substances, and determining the optimal concentration of a melatonin culture medium which is processed by the optimal anthocyanin accumulation amount, the expression amount of the anthocyanin metabolic genes and the content of the flavonoid substances correspondingly as the optimal concentration of the melatonin culture medium for promoting the accumulation of the flavonoid substances in the leaf tissues of the ornamental crabapple;
and performing propagation culture on the tissue culture seedling of the ornamental crabapple leaf variety by using the determined melatonin culture medium with the optimal concentration.
In the method, the tissue culture seedling of the ornamental crabapple leaf variety is a subculture seedling which grows for 20-30 days. Preferably, the tissue culture seedling of the ornamental crabapple leaf variety is a tissue culture seedling of 'royal' of the ornamental crabapple normal color purple leaf variety, namely a subculture seedling growing for 20-30 days.
In the method, the melatonin culture media with different concentrations are as follows:
melatonin media at concentrations of 0. mu.g/l, 50. mu.g/l, 100. mu.g/l, 200. mu.g/l, 500. mu.g/l, 1000. mu.g/l, respectively.
In the above method, the determination of the optimal concentration of melatonin medium is: melatonin medium at a concentration of 200. mu.g/l.
In the above method, the melatonin medium is: MS culture medium with concentration of 4.74g/L, 6-BA solution with concentration of 1mg/L, NAA solution with concentration of 0.05mg/L, sucrose with concentration of 30g/L and agar with concentration of 7 g/L.
In the melatonin culture medium of the method, the 6-BA solution is a 6-Benzyl amino urea solution, the mother solution is 1mg/mL, and 2000 microliters per liter is added;
the NAA solution is a Naphtylacetic acid solution, the mother solution is 1mg/mL, and 400 microliters per liter is added.
In the above method, the melatonin culture medium is prepared, and then sterilized under high pressure for use.
In the method, the sampling mode of the blade is as follows: only the leaf parts were removed with scissors tweezers and collected in a 50ml centrifuge tube and frozen in liquid nitrogen.
In the method, the tissue culture seedling material is obtained by the following culture modes:
subculturing ornamental Chinese flowering crabapple leaf varieties, and placing the seedlings on a tissue culture shelf; the culture conditions are as follows: the relative humidity is 60-70%, the temperature is 24-26 ℃, the illumination time is 16h illumination/8 h dark light cycle, the illumination intensity is 1500-2000 Lx, and the tissue culture seedlings are obtained after 15 days of culture.
The embodiments of the present invention are described in further detail below.
The embodiment provides a method for rapidly promoting the accumulation of flavonoid substances in ornamental crabapple leaf tissues by using melatonin with specific concentration, which comprises the following steps:
selecting tissue culture seedlings of ornamental malus spectabilis normal purple leaf variety 'Wang' as materials, selecting seedlings with consistent growth vigor and vigorous buds for 20-25 days, subculturing the seedlings with good leaves on a culture medium added with 0 mug/l, 50 mug/l, 100 mug/l, 200 mug/l, 500 mug/l and 1000 mug/l, and then carrying out normal illumination culture for 2 weeks;
selecting leaves with obvious phenotype and good growth vigor for sample storage, subsequently extracting RNA, further converting the RNA into cDNA for qRT-PCR experiment, and detecting the transcription level of key structural genes in an anthocyanin metabolic pathway.
In the method, the used subculture seedlings are subcultured seedlings which grow for 20-30 days for ornamental malus spectabilis normal color purple leaf variety 'Wang nationality'.
In the above method, the melatonin culture medium used is: MS (Murashige and Skoog medium; product of chembase, 4.74g per liter) +1 mg/L6-BA solution (1 mg/mL of 6-Benzyl amino urea mother liquor, 2000. mu.L per liter) +0.05mg/L NAA solution (1 mg/mL of Naphtylacetic acid mother liquor, 400. mu.L per liter) +30g/L sucrose +7g/L agar) is prepared, autoclaved, and then melatonin (0. mu.g/L, 50. mu.g/L, 100. mu.g/L, 200. mu.g/L, 500. mu.g/L, 1000. mu.g/L) is added.
In the above method, the sampling mode: only the leaf parts were removed with scissors tweezers and collected in a 50ml centrifuge tube and frozen in liquid nitrogen.
In the above method, two conditions of cultivation: and (3) light treatment: placing the seedlings on a tissue culture room frame after subculture; the cells were cultured at 23 ℃ for 16 hours under light for 15 days.
Aiming at the phenomenon of progenitor of the tissue culture seedling of the woody plant, namely the ornamental crabapple, caused by more subculture times, the content of flavonoid substances in leaf tissues of the ornamental crabapple can be promoted by screening out melatonin with specific concentration, so that the leaves are red. The phenomenon that the leaves of the subculture tissue culture seedlings return to ancestors due to excessive generation is well solved.
The method screens out the melatonin with specific concentration of 200 mu g/l, subcultures the tissue culture seedlings to a culture medium with the melatonin, cultures the tissue culture seedlings for 15 days by using two culture conditions, observes the phenotype, samples and extracts RNA, reverses the cDNA to express, and finally determines the transcription level of genes related to the melatonin metabolic pathway and the transcription level of genes of key structures related to the anthocyanin metabolic pathway so as to determine that the melatonin with the concentration of 200 mu g/l can influence the metabolism of the endogenous melatonin of the plant and can influence the genes related to the anthocyanin metabolism.
Example (b):
the embodiment provides a method for promoting accumulation of flavonoid substances in ornamental begonia leaf tissues, which is a method capable of rapidly promoting color and luster of ornamental begonia leaves and comprises the following steps:
ordinary MS medium: MS (Murashige and Skoog medium; product of chembase, 4.74g per liter) +1 mg/L6-BA solution (1 mg/mL of 6-Benzyl amino urea mother liquor, 2000. mu.L per liter) +0.05mg/L NAA solution (1 mg/mL of Naphtylacetic acid mother liquor, 400. mu.L per liter) +30g/L sucrose +7g/L agar) is prepared, autoclaved, and then melatonin (0. mu.g/L, 50. mu.g/L, 100. mu.g/L, 200. mu.g/L, 500. mu.g/L, 1000. mu.g/L) is added. Culturing in a normal illumination culture rack for 15 days;
step 2, observation of phenotype, fig. 1 shows that different concentrations of melatonin were used in the present invention:
step 3, culturing the Chinese flowering crabapple seedlings treated by different concentrations of melatonin according to the phenotypes, selecting leaves with obvious phenotypes, storing the leaves at-80 ℃ for later use, detecting the content of flavonoid substances, and selecting the optimal melatonin concentration according to the anthocyanin accumulation amount and the anthocyanin metabolism gene expression amount of the leaves of the tissue culture seedlings treated by various concentrations, which is shown in figure 2;
step 4, qRT-PCR is carried out to detect the expression of melatonin pathway related genes and anthocyanin metabolic pathway related genes (see figure 3);
in the above method, the transcription levels of other structural genes for anthocyanin metabolism in the 'royal' leaf tissue after different concentrations of melatonin treatment are shown in fig. 3, and the expression levels of transcription factors for anthocyanin metabolism in the 'royal' leaf tissue after different concentrations of melatonin treatment are shown in fig. 4.
The method takes the 'royal' of the ornamental crabapple normal color purple leaf variety as a material, melatonin treatment ornamental crabapple tissue culture seedlings with different concentration gradients are designed, 200 mu g/l of melatonin treatment 'royal' is screened out according to phenotypes, purple red of leaves is very obvious, the color of the leaves with high concentration or low concentration is not obviously changed, the content of flavonoid substances in the 'royal' leaves treated by 200 mu g/l of melatonin, the transcription level of key genes in a flavonoid metabolic pathway and the transcription level of genes related to a melatonin pathway are detected subsequently, and the result shows that the anthocyanin content is highest in the 'royal' treated by 200 mu g/l of melatonin. The invention provides a method for improving the progenitor phenomenon of the 'Wang' due to the increase of the subculture frequency, and further provides a good theoretical basis for the genetic improvement of the color-leaf plants. It can be seen that the method of the invention is applicable to ornamental begonia leaves of various varieties.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are included in the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (7)
1. A method for promoting accumulation of flavonoid substances in ornamental crabapple leaf tissues is characterized by comprising the following steps:
taking a tissue culture seedling of an ornamental crabapple leaf variety as a material, carrying out propagation on the tissue culture seedling of the variety in melatonin culture media respectively added with melatonin of different concentrations, and then carrying out culture for 2 weeks under normal illumination, wherein the different concentrations of the melatonin are respectively as follows: 0. mu.g/l, 50. mu.g/l, 100. mu.g/l, 200. mu.g/l, 500. mu.g/l, 1000. mu.g/l;
selecting leaf storage samples with obvious phenotype and good growth vigor, extracting RNA from the storage sample leaves, inverting the RNA into cDNA to perform a qRT-PCR experiment, detecting the transcription level of key structural genes in an anthocyanin metabolic pathway, detecting the content of flavonoid substances, and determining the optimal melatonin concentration which is processed by the optimal anthocyanin accumulation amount, the expression amount of the anthocyanin metabolic genes and the content of the flavonoid substances as the optimal concentration for promoting the accumulation of the flavonoid substances in the leaf tissues of the ornamental crabapples, wherein the optimal concentration is 200 mug/l;
the melatonin culture medium comprises the following components except melatonin: MS culture medium with concentration of 4.74g/L, 6-BA solution with concentration of 1mg/L, NAA solution with concentration of 0.05mg/L, sucrose with concentration of 30g/L and agar with concentration of 7 g/L;
and performing propagation culture on the tissue culture seedling of the ornamental crabapple leaf variety by using the determined melatonin culture medium with the optimal concentration.
2. The method for promoting accumulation of flavonoid substances in ornamental crabapple leaf tissues according to claim 1, wherein the tissue culture seedlings of the ornamental crabapple leaf varieties are subcultured seedlings of the ornamental crabapple leaf varieties growing for 20-30 days.
3. The method for promoting accumulation of flavonoid substances in leaf tissues of ornamental crabapple as claimed in any one of claims 1 to 2, wherein in the melatonin culture medium, the 6-BA solution is a 6-Benzyl Aminopurine solution, the mother liquor is 1mg/mL, and 2000 microliters per liter is added;
the NAA solution is a Naphtylacetic acid solution, the mother solution is 1mg/mL, and 400 microliters per liter is added.
4. The method for promoting accumulation of flavonoid substances in leaf tissue of ornamental crabapple according to any one of claims 1 to 2, wherein the melatonin culture medium is prepared and then used after autoclaving.
5. The method for promoting accumulation of flavonoid substances in ornamental begonia leaf tissue according to any one of claims 1 to 2, wherein said leaf is sampled by: only the leaf parts were removed with scissors tweezers and collected in a 50ml centrifuge tube and frozen in liquid nitrogen.
6. The method for promoting accumulation of flavonoid substances in ornamental crabapple leaf tissues according to any one of claims 1 to 2, wherein the tissue culture seedling materials are obtained by the following culture modes:
subculturing ornamental Chinese flowering crabapple leaf varieties, and placing the seedlings on a tissue culture shelf; the culture conditions are as follows: the relative humidity is 60-70%, the temperature is 24-26 ℃, the illumination time is 16h illumination/8 h dark light cycle, the illumination intensity is 1500-2000 Lx, and the tissue culture seedlings are obtained after 15 days of culture.
7. The method for promoting accumulation of flavonoid substances in ornamental begonia leaf tissues according to any one of claims 1 to 2, wherein in the method, the ornamental begonia leaf variety is an ornamental begonia normal-color purple leaf variety.
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| CN113906945B (en) * | 2021-09-23 | 2023-04-18 | 中南林业科技大学 | Method for promoting accumulation of flavonoid content in camellia oleifera seeds |
| CN114080946B (en) * | 2021-10-09 | 2023-03-31 | 北京林业大学 | Application of exogenous melatonin in improving salt tolerance of ginkgo and increasing content of ginkgo leaf flavone |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1288656A (en) * | 2000-11-09 | 2001-03-28 | 中国科学院石家庄农业现代化研究所 | Fast seedling growing mehtod for Chinese flowering crabapple zhumei |
| CN1794907A (en) * | 2003-05-22 | 2006-06-28 | 株式会社鹿児岛Tlo | Method of crossing flower color genotypes |
| CN101258245A (en) * | 2005-08-30 | 2008-09-03 | 新西兰园艺和食品研究院有限公司 | Compositions and methods for modulating pigment production in plants |
| CN103392597A (en) * | 2013-07-24 | 2013-11-20 | 江苏绿苑园林建设有限公司 | Tissue culture method of North American begonia |
| CN104115747A (en) * | 2014-07-03 | 2014-10-29 | 山东农业大学 | Production of apple flavonoids by using tissue culture method |
| CN104686005A (en) * | 2015-02-04 | 2015-06-10 | 中国农业大学 | Method for increasing content of anthocyanin in purple cabbage |
| CN107258538A (en) * | 2017-07-01 | 2017-10-20 | 西北农林科技大学 | A kind of apple rootstock SH38 rapid propagation methods |
-
2018
- 2018-04-27 CN CN201810394671.9A patent/CN108849498B/en not_active Expired - Fee Related
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1288656A (en) * | 2000-11-09 | 2001-03-28 | 中国科学院石家庄农业现代化研究所 | Fast seedling growing mehtod for Chinese flowering crabapple zhumei |
| CN1794907A (en) * | 2003-05-22 | 2006-06-28 | 株式会社鹿児岛Tlo | Method of crossing flower color genotypes |
| CN101258245A (en) * | 2005-08-30 | 2008-09-03 | 新西兰园艺和食品研究院有限公司 | Compositions and methods for modulating pigment production in plants |
| CN103392597A (en) * | 2013-07-24 | 2013-11-20 | 江苏绿苑园林建设有限公司 | Tissue culture method of North American begonia |
| CN104115747A (en) * | 2014-07-03 | 2014-10-29 | 山东农业大学 | Production of apple flavonoids by using tissue culture method |
| CN104686005A (en) * | 2015-02-04 | 2015-06-10 | 中国农业大学 | Method for increasing content of anthocyanin in purple cabbage |
| CN107258538A (en) * | 2017-07-01 | 2017-10-20 | 西北农林科技大学 | A kind of apple rootstock SH38 rapid propagation methods |
Non-Patent Citations (6)
| Title |
|---|
| A label-free differential proteomics analysis reveals the effect of melatonin on promoting fruit ripening and anthocyanin accumulation upon postharvest in tomato;Qianqian Sun等;《Journal of Pineal Research》;20161231;第61卷(第2期);第138-153页 * |
| Application of melatonin promotes anthocyanin accumulation in crabapple leaves;Li Chen等;《Plant Physiology and Biochemistry》;20191231;第142卷;第332–341页 * |
| Melatonin Improved Anthocyanin Accumulation by Regulating Gene Expressions and Resulted in High Reactive Oxygen Species Scavenging Capacity in Cabbage;Na Zhang等;《Frontiers in PlantScience》;20161231;第7卷;第1-17页 * |
| 植物激素对类黄酮代谢调控机制研究进展;于江珊等;《中国中药杂志》;20210831;第46卷(第15期);第3806-3813页 * |
| 褪黑素对光暗培养条件下紫锥菊毛状根生长和代谢的影响;何珊珊;《中国科学院机构知识库网格》;20130923;摘要 * |
| 观叶秋海棠叶片微繁方法的研究;杜文文等;《现代园林》;20151231;第12卷(第4期);第266页 * |
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