CN113367137A - Culture solution for rapidly inducing tobacco leaf aging and treatment method - Google Patents
Culture solution for rapidly inducing tobacco leaf aging and treatment method Download PDFInfo
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- CN113367137A CN113367137A CN202110755360.2A CN202110755360A CN113367137A CN 113367137 A CN113367137 A CN 113367137A CN 202110755360 A CN202110755360 A CN 202110755360A CN 113367137 A CN113367137 A CN 113367137A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
- A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
- A01N43/14—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings
- A01N43/16—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings with oxygen as the ring hetero atom
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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
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/72—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with nitrogen atoms and oxygen or sulfur atoms as ring hetero atoms
- A01N43/84—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with nitrogen atoms and oxygen or sulfur atoms as ring hetero atoms six-membered rings with one nitrogen atom and either one oxygen atom or one sulfur atom in positions 1,4
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Abstract
The invention provides a culture solution for rapidly inducing leaf senescence, belonging to the technical field of plant production, wherein the culture solution comprises 10-60 g/L of sucrose, and tobacco plants are treated by the culture solution containing the sucrose, so that the accumulation of tobacco biomass is promoted, the senescence of tobacco leaves is accelerated, the maturation time is shortened, the labor intensity is greatly reduced, and the industrial value is improved.
Description
Technical Field
The invention relates to the technical field of plant production, in particular to a culture solution for rapidly inducing tobacco leaf senescence and a treatment method.
Background
The senescence of plant leaves, which is the last stage of plant growth and development, is influenced by various factors such as complex exogenous environmental signals, endogenous plant hormones, leaf age and the like. The leaf senescence process is a programmed process which influences the recovery and recycling of substances, and the speed of the plant senescence process obviously influences the benefits of agricultural production, such as the yield and the quality of crops.
Tobacco is one of the major leaf-based commercial crops and has both smoking and medicinal value. China is the country with the largest tobacco planting area and the largest tobacco yield, and makes great contribution to the social and economic development of China. The leaf is the product organ of tobacco, and the goal of tobacco production is to obtain high-yield and high-quality tobacco leaves. The aging and yellowing are the last stage before the tobacco leaves are harvested, the chlorophyll content is reduced in the period, the photosynthesis of the leaves is weakened, and the decomposition of protein, nucleic acid and lipid is realized to promote the formation of the quality of the leaves. However, in actual production, the problem of early aging of the tobacco leaves always exists, so that the labor intensity of tobacco growers for harvesting the tobacco leaves is high, and the tobacco leaf harvesting time is long. Therefore, on the premise of ensuring the biomass of tobacco flakes, the tobacco leaf senescence is quickly induced, the labor intensity of tobacco growers is relieved, the tobacco leaf harvesting time can be shortened, and the economic benefit is improved, but other reports related to the induction of the leaf senescence on tobacco are not seen yet.
Therefore, it is an urgent technical problem to provide a culture solution and a treatment method for rapidly inducing tobacco leaf aging.
Disclosure of Invention
The invention aims to provide a culture solution for rapidly inducing tobacco leaf senescence, which can promote the accumulation of biomass of tobacco leaves and shorten the maturation time.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a culture solution for rapidly inducing tobacco leaf senescence, which comprises 10-60 g/L of sucrose.
Preferably, the culture solution further comprises 1/2MS 2-2.5 g/L, MES 0.1.1-0.8 g/L.
Preferably, the culture solution comprises the following components in mass concentration: 1/2MS 2.2-2.4 g/L, sucrose 15-30 g/L, MES 0.3-0.7 g/L.
Preferably, the pH of the culture broth is 5.8.
The invention also provides a processing method for rapidly inducing the tobacco leaf aging, which comprises the following steps:
(1) selecting tobacco seedlings with consistent growth vigor and size;
(2) and (3) carrying out spraying treatment on tobacco leaves by using any culture solution or taking leaves to disperse in any culture solution.
Preferably, the spraying treatment mode is spraying once a day for 1-9 days continuously; the time for taking the leaves to disperse in the culture solution is 1-9 days.
Preferably, the tobacco is a 5-leaf, 1-heart plant.
Preferably, the tobacco comprises cut tobacco leaves or field grown tobacco plants.
Preferably, the tobacco is tobacco K326.
Preferably, the spraying ensures the leaves are moist.
Compared with the prior art, the technical scheme of the invention has the beneficial effects that:
(1) according to the invention, the tobacco plants are treated by the culture solution containing sucrose, so that the accumulation of tobacco biomass is promoted, the aging of tobacco is accelerated, the maturation time is shortened, the labor intensity is greatly reduced, and the industrial value is improved.
(2) The treatment method is simple and easy to operate, and the culture solution for inducing the tobacco leaf aging can be quickly prepared.
Drawings
FIG. 1 is a graph of the color change of leaf disks of examples 1-4 on days 1, 3, 5 and 7;
FIG. 2 is a graph showing the results of chlorophyll-a contents of leaf disks of examples 1 to 4 on days 1, 3, 5, 7 and 9;
FIG. 3 is a graph showing the results of biomass accumulation on days 1, 3, 5, 7 and 9 in the leaf disks of examples 1 to 4.
Detailed Description
The invention provides a culture solution for rapidly inducing tobacco leaf senescence, which comprises 10-60 g/L of sucrose.
In the invention, the culture solution also preferably comprises 1/2MS 2-2.5 g/L, MES 0.1-0.8 g/L; the culture solution preferably comprises the following components in mass concentration: 1/2MS 2.2-2.4 g/L, sucrose 15-30 g/L, MES 0.3-0.7 g/L; wherein said MS is free of agar and sucrose; the pH of the culture broth is preferably 5.8. In the invention, the culture solution has the effects of promoting the rapid senescence of tobacco leaves and improving the accumulation of the biomass of the tobacco leaves. The preparation method of the culture solution comprises the steps of dissolving sucrose, MS and MES which are weighed according to the mass concentration by using ultrapure water, then adjusting the pH value of the solution to 5.8, using the ultrapure water to perform volume fixing on the solution after the pH value is adjusted, pouring the liquid after the volume fixing into a sterilization bottle for sealing, placing the sealed solution into an autoclave for sterilization at 120 ℃ for 30 minutes, and cooling the sterilized solution to room temperature to obtain the culture solution. The MS, MES and sucrose of the present invention were all purchased from Solambio.
The invention also provides a treatment method for rapidly inducing leaf senescence, which comprises the following steps:
(1) selecting tobacco seedlings with consistent growth vigor and size;
(2) and (3) carrying out spraying treatment on tobacco leaves by using any culture solution or taking leaves to disperse in any culture solution.
In the present invention, the tobacco leaves are subjected to spraying treatment with any of the above-mentioned culture solutions. The spraying treatment mode is preferably one time of spraying every day for 1-9 days. The tobacco is preferably a 5-leaf and 1-heart plant. The tobacco preferably comprises excised leaf or field grown tobacco plants. The tobacco is preferably tobacco K326. The spraying is preferably carried out in such a way that the leaves are kept moist. The time for taking the leaves to disperse in the culture solution is 1-9 days. The tobacco K326 is provided by a national crop germplasm resource platform and a tobacco germplasm resource sub-platform, and the storage unit is the tobacco research institute of Chinese academy of agricultural sciences.
The treatment method is simple and rapid to operate, can rapidly induce the aging of the tobacco leaves, and shortens the maturation time.
The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
The culture solution for rapidly inducing the tobacco leaf aging consists of the following components in mass concentration: 10.27g/L of sucrose, 1/2MS 2.37g/L, MES 0.5.5 g/L, wherein the pH value of the culture solution is 5.8.
The treatment method for rapidly inducing leaf senescence of the embodiment comprises the following steps:
(1) selecting tobacco seedlings with consistent growth vigor and size;
(2) spraying 5-leaf 1-core tobacco K326 leaves with the culture solution or taking the leaves to disperse in the culture solution, wherein the spraying treatment mode is to spray once every morning and continuously spray for 9 days, and each leaf is guaranteed to be wet when spraying; the time period for which the leaves were dispersed in the culture solution was 9 days.
Example 2
The culture solution for rapidly inducing the tobacco leaf aging consists of the following components in mass concentration: 20.54g/L of sucrose, 1/2MS 2.37g/L, MES 0.5.5 g/L, wherein the pH value of the culture solution is 5.8.
The treatment method for rapidly inducing leaf senescence of the embodiment comprises the following steps:
(1) selecting tobacco seedlings with consistent growth vigor and size;
(2) spraying 5-leaf 1-core tobacco K326 leaves with the culture solution or taking the leaves to disperse in the culture solution, wherein the spraying treatment mode is to spray once every morning and continuously spray for 9 days, and each leaf is guaranteed to be wet when spraying; the time period for which the leaves were dispersed in the culture solution was 9 days.
Example 3
The culture solution for rapidly inducing the tobacco leaf aging consists of the following components in mass concentration: 30.81g/L of sucrose, 1/2MS 2.37g/L, MES 0.5.5 g/L, wherein the pH value of the culture solution is 5.8.
The treatment method for rapidly inducing leaf senescence of the embodiment comprises the following steps:
(1) selecting tobacco seedlings with consistent growth vigor and size;
(2) spraying 5-leaf 1-core tobacco K326 leaves with the culture solution or taking the leaves to disperse in the culture solution, wherein the spraying treatment mode is to spray once every morning and continuously spray for 9 days, and each leaf is guaranteed to be wet when spraying; the time period for which the leaves were dispersed in the culture solution was 9 days.
Example 4
The culture solution for rapidly inducing the tobacco leaf aging consists of the following components in mass concentration: 41.08g/L of sucrose, 1/2MS 2.37g/L, MES 0.5.5 g/L, wherein the pH value of the culture solution is 5.8.
The treatment method for rapidly inducing leaf senescence of the embodiment comprises the following steps:
(1) selecting tobacco seedlings with consistent growth vigor and size;
(2) spraying 5-leaf 1-core tobacco K326 leaves with the culture solution or taking the leaves to disperse in the culture solution, wherein the spraying treatment mode is to spray once every morning and continuously spray for 9 days, and each leaf is guaranteed to be wet when spraying; the time period for which the leaves were dispersed in the culture solution was 9 days.
Example 5
The culture solution for rapidly inducing the tobacco leaf aging consists of the following components in mass concentration: sucrose 51.35g/L, 1/2MS 2.37g/L, MES 0.5.5 g/L, wherein the pH value of the culture solution is 5.8.
The treatment method for rapidly inducing leaf senescence of the embodiment comprises the following steps:
(1) selecting tobacco seedlings with consistent growth vigor and size;
(2) spraying 5-leaf 1-core tobacco K326 leaves with the culture solution or taking the leaves to disperse in the culture solution, wherein the spraying treatment mode is to spray once every morning and continuously spray for 9 days, and each leaf is guaranteed to be wet when spraying; the time period for which the leaves were dispersed in the culture solution was 9 days.
Example 6
The culture solution for rapidly inducing the tobacco leaf aging consists of the following components in mass concentration: 10g/L of sucrose, 1/2MS 2.5g/L, MES 0.8.8 g/L, wherein the pH value of the culture solution is 5.8.
The treatment method for rapidly inducing leaf senescence of the embodiment comprises the following steps:
(1) selecting tobacco seedlings with consistent growth vigor and size;
(2) spraying 5-leaf 1-core tobacco K326 leaves with the culture solution or taking the leaves to disperse in the culture solution, wherein the spraying treatment mode is to spray once every morning and continuously spray for 5 days, and each leaf is guaranteed to be wet when spraying; the time period for which the leaves were dispersed in the culture solution was 5 days.
Example 7
The culture solution for rapidly inducing the tobacco leaf aging consists of the following components in mass concentration: 60g/L of sucrose, 1/2MS 2g/L, MES 0.1.1 g/L of sucrose, wherein the pH value of the culture solution is 5.8.
The treatment method for rapidly inducing leaf senescence of the embodiment comprises the following steps:
(1) sowing in a tray containing a mixture of 1:1 peat and vermiculite (V/V), and selecting tobacco seedlings with consistent growth and size;
(2) spraying 5-leaf 1-core tobacco K326 leaves with the culture solution or taking the leaves to disperse in the culture solution, wherein the spraying treatment mode is to spray once every morning and continuously spray for 3 days, and each leaf is guaranteed to be wet when spraying; the time period for which the leaf was dispersed in the culture solution was 3 days.
Example 8
In the in vitro leaf senescence induction test according to the manner of example 1-4, in order to better reflect the test result, the leaf in the step (2) of example 1-4 is taken, the hole is punched, the leaf disc is taken and dispersed in a culture dish containing a culture solution, and the rest steps are completely the same. In this example, the leaf disks of examples 1 to 4 and the control group of 1, 3, 5 and 7 days were photographed and analyzed by dispersing the leaf disks in a petri dish containing ultrapure water for 9 days as the control group, and the results are shown in FIG. 1.
From the results in FIG. 1, it is clear that the culture solutions of examples 1 to 4 and the treatment method for inducing leaf senescence showed that the leaf yellowing time was faster than that of the control group, indicating that the culture solution of the present invention promotes the degradation of chlorophyll.
Example 9
In the in vitro leaf senescence induction test according to the manner of example 1-4, in order to better reflect the test result, the leaf in the step (2) of example 1-4 is punched, leaf discs are taken and dispersed in a culture dish containing a culture solution, and the rest steps are completely the same. In the embodiment, the leaf discs are taken and dispersed in a culture dish containing ultrapure water for 9 days to serve as a control group, and samples are taken on 1 st, 3 rd, 5 th, 7 th and 9 th days of induction to measure the content of chlorophyll a;
the method for measuring the content of chlorophyll a comprises the following steps: to ensure reproducibility of the measurements, at least three biological replicates per dish were performed to minimize experimental error. Chlorophyll a content was determined according to the method of (ZHao L, Xia Y, Wu XY, Schippers JHM, lacing HC (2018) photopic analysis and molecular markers of leaf sensors. methods Mol Biol 1744: 35-48). The leaf discs are weighed by an electric balance before being soaked in absolute ethyl alcohol. 10mL of absolute ethanol was added to the centrifuge tube. The leaves are completely immersed into a 10mL centrifuge tube and stored at room temperature for 24-48 hours in the dark. The aluminum foil paper is wrapped on the centrifuge tube to completely decolorize the leaves. Blank solution was used as control. The absorbance values of the two solutions were calculated at 665nm and 649nm, respectively, using an M200PRO spectrophotometer (TECAN), and the data were recorded. The calculation formula of the chlorophyll a content of the leaves is as follows: c (mg/g) ═ Ca + Cb × V/W × g, where Ca ═ 13.95 × X1-6.88Y × Y1, Cb ═ 24.96 × Y1-7.32 × X1, V ═ 10mL, W is the fresh weight of the leaves, and X1 and Y1 are absorbance values at 665nm and 649nm, respectively. The results of the chlorophyll a content of the leaf disks of examples 1 to 4 after induction for days 1, 3, 5, 7 and 9 are shown in FIG. 2, wherein Chl a in FIG. 2 represents the green color a of the leaf.
As can be seen from FIG. 2, the content of chlorophyll a in the control group has no significant difference with the time, the content of chlorophyll a in examples 1 to 4 gradually decreases, and the content of chlorophyll a gradually decreases with the increase of the sucrose concentration in the culture solution, which indicates that the culture solution of the present invention promotes the aging of the tobacco leaves.
The fresh weight of the tobacco leaf discs induced by examples 1-4 was weighed, and the weighing results are shown in fig. 3.
As is clear from FIG. 3, the biomass accumulation of leaves was higher in examples 1 to 4 on the same days than in the control group; the accumulation of the biomass of the tobacco leaves is gradually increased along with the increase of the concentration of the sucrose in the culture solution, which shows that the culture solution improves the accumulation of the biomass of the tobacco leaves.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. A culture solution for rapidly inducing tobacco leaf senescence is characterized by comprising 10-60 g/L of sucrose.
2. The culture solution of claim 1, further comprising 1/2MS 2-2.5 g/L, MES 0.1-0.8 g/L.
3. The culture solution according to claim 2, wherein the culture solution comprises the following components in mass concentration:
1/2MS 2.2-2.4 g/L, sucrose 15-30 g/L, MES 0.3-0.7 g/L.
4. The culture solution according to any one of claims 1 to 3, wherein the pH of the culture solution is 5.8.
5. A treatment method for rapidly inducing the aging of tobacco leaves, which is characterized by comprising the following steps:
(1) selecting tobacco seedlings with consistent growth vigor and size;
(2) spraying tobacco leaves with the culture solution of any one of claims 1 to 4 or dispersing the tobacco leaves in the culture solution of any one of claims 1 to 4.
6. The treatment method according to claim 5, wherein the spraying treatment is carried out by spraying once a day for 1-9 days; the time for taking the leaves to disperse in the culture solution is 1-9 days.
7. The treatment method according to claim 5, wherein the tobacco is a 5-leaf 1-heart plant.
8. The treatment method of claim 5, wherein the tobacco comprises cut tobacco leaves or field grown tobacco plants.
9. The treatment according to claim 5, wherein the tobacco is tobacco K326.
10. The treatment method according to claim 5, wherein said spraying ensures the wetting of the leaves.
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