CN115720775B - Method for promoting quick yellowing of Wucai - Google Patents
Method for promoting quick yellowing of Wucai Download PDFInfo
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Abstract
The invention relates to the technical field of vegetable cultivation, and provides a method for promoting quick yellowing of Wucai, which comprises the following steps of performing low-temperature pretreatment on Wucai plants when reaching a plant period: 0:00-7:30,8 ℃;7:30-10:30, 11 ℃;10:30-14:30, 16 ℃;14:30-19:30, 11 ℃;19:30-24:00,8 ℃; after 2 days of low-temperature pretreatment, respectively regulating the temperature of each time period to 6 ℃ downwards for continuous culture; at the same time, the Wu Cai inner leaf was sprayed with 1mM MeJA. The invention jointly regulates and controls the yellowing effect of the black vegetable leaves through the combined action mode of low-temperature gradient pretreatment, formal low-temperature gradient treatment and She Penshi MeJA in the black vegetable, so that the black vegetable leaves can be quickly yellowing, and the black vegetable with ornamental and edible values is obtained, thereby providing a powerful foundation for the application and development of the black vegetable.
Description
Technical Field
The invention relates to the technical field of vegetable cultivation, in particular to a method for promoting quick yellowing of black vegetables.
Background
Wucai (Brassica campestris L.) is a subspecies of Chinese cabbage, native to China, and mainly distributed in river basin, and is a nutritious vegetable rich in minerals and vitamins, and leaves are colorful. Among the existing black vegetables, some black vegetables, such as W7-2 black vegetables, have green leaves in the seedling stage at the early autumn temperature (24-33 ℃), keep green at normal temperature in the plant-forming stage, and have yellow leaves along with the growth of plants at the low temperature (8-18 ℃). At this time, although She Bianhuang is contained, the phenotype can still keep growing in winter, and the yellow black-bone subjected to frosted snow has the characteristics of high ornamental value, less crude fiber, good taste and the like.
Therefore, if a method for promoting the quick yellowing of the black vegetables can be researched, a powerful foundation is provided for the application development of the black vegetables.
Disclosure of Invention
The invention aims to provide a method for promoting quick yellowing of Wucai.
The invention adopts the following technical scheme to solve the technical problems:
a method for promoting quick yellowing of Wucai plants, when the Wucai plants reach the plant period, the Wucai plants are subjected to low-temperature pretreatment at the following temperature: 0:00-7:30,8 ℃;7:30-10:30, 11 ℃;10:30-14:30, 16 ℃;14:30-19:30, 11 ℃;19:30-24:00,8 ℃;
after 2 days of low-temperature pretreatment, respectively regulating the temperature of each time period to 6 ℃ downwards for continuous culture; at the same time, the Wu Cai inner leaf was sprayed with 1mM MeJA.
As one preferable mode of the invention, 300-350 mu l of the spray is continuously sprayed into the Wucai She Penshi MeJA three times.
As one of the preferable modes of the invention, the method comprises the following specific steps:
(1) Sowing the black vegetable seeds in a container, and culturing the seeds;
(2) Transplanting after the seeds germinate and have 4 true leaves;
(3) When the black vegetable plants reach the plant period, placing the black vegetable plants in an ultralow temperature incubator for low-temperature pretreatment, wherein the pretreatment temperature is as follows: 0:00-7:30,8 ℃;7:30-10:30, 11 ℃;10:30-14:30, 16 ℃;14:30-19:30, 11 ℃;19:30-24:00,8 ℃;
(4) After 2 days of low-temperature pretreatment, the temperature of the ultralow-temperature incubator is adjusted to 6 ℃ for continuous culture, namely, the temperature is 0:00-7:30 and 2 ℃;7:30-10:30,5 ℃;10:30-14:30, 10 ℃;14:30-19:30,5 ℃;19:30-24:00,2 ℃; simultaneously, spraying treatment is carried out on the internal leaves of the Wucai with 1mM MeJA, and the spraying treatment is carried out three times continuously, wherein 300-350 mu l of the internal leaves of the Wucai are sprayed each time.
In a preferred embodiment of the present invention, in the step (1), the culture environment of the black vegetable seeds is: day temperature 25+ -1 deg.C, night temperature 18+ -1 deg.C, light quantum flux density 300-500 mu mol.m -2 ·s -1 The relative humidity is 60-70%.
In the step (2), transplanting the black vegetable seedlings to nutrient soil: vermiculite = 3:1 in plastic pots.
As a preferred embodiment of the present invention, in the step (3), the irradiation intensity is set to 300. Mu. Mol m during the low-temperature pretreatment -2 s -1 。
As a preferred embodiment of the present invention, in the step (4), the illumination intensity is set to 300. Mu. Mol m when the cultivation is performed by adjusting the temperature to 6℃ -2 s -1 。
Compared with the prior art, the invention has the advantages that: the invention jointly regulates and controls the yellowing effect of the black vegetable leaves through the combined action mode of low-temperature gradient pretreatment, formal low-temperature gradient treatment and She Penshi MeJA in the black vegetable, so that the black vegetable leaves can be quickly yellowing, and the black vegetable with ornamental and edible values is obtained, thereby providing a powerful foundation for the application and development of the black vegetable.
Drawings
FIG. 1 is a diagram of Wucai varieties with different degrees of yellowing in Experimental example 1;
FIG. 2 is a phenotype chart of W7-2 black vegetables under different treatments in Experimental example 5 (in the figure, LT+Control represents "low temperature Control", LT+MeJA represents "exogenous MeJA treatment under low temperature", LT+DIECA represents "DIECA treatment under low temperature", NT+Control represents "normal temperature Control", NT+MeJA represents "exogenous MeJA treatment under normal temperature", NT+DIECA represents "DIECA treatment under normal temperature");
FIG. 3 is the chlorophyll change of W7-2 leaf after MeJA treatment at low temperature in experimental example 6 (in the figure, mock represents a fresh water control);
FIG. 4 is a graph showing the change in a-value of W7-2 leaves after MeJA treatment at low temperature in experimental example 6 (in the graph, mock represents a fresh water control);
FIG. 5 is a graph showing the change in b-value of W7-2 leaves after MeJA treatment at low temperature in experimental example 6 (in the graph, mock represents a fresh water control);
FIG. 6 is a graph showing the change in JA content of W7-2 leaves after MeJA treatment at low temperature in experimental example 6 (in the graph, mock represents a fresh water control).
Detailed Description
The following describes in detail the examples of the present invention, which are implemented on the premise of the technical solution of the present invention, and detailed embodiments and specific operation procedures are given, but the scope of protection of the present invention is not limited to the following examples.
Example 1
The method for promoting the quick yellowing of the black vegetables in the embodiment comprises the following steps of:
(1) The black vegetable seeds are sown in containers and placed in the following environments for cultivation: day temperature 24 ℃, night temperature 17 ℃, and light quantum flux density 300 mu mol.m -2 ·s -1 The relative humidity is 60%.
(2) After germination of the seeds and 4 true leaves, transplanting the seedlings to nutrient soil: vermiculite = 3:1 in plastic pots.
(3) When the black vegetable plant reaches the plant period, the black vegetable plant is placed in an ultralow temperature incubator for low-temperature pretreatment, the temperature is set to 8 ℃ (0:00-7:30), 11 ℃ (7:30-10:30), 16 ℃ (10:30-14:30), 11 ℃ (14:30-19:30), 8 ℃ (19:30-24:00), and the illumination intensity is 300 mu mol m -2 s -1 。
(4) After 2 days of low temperature pretreatment, the temperature of the ultra-low temperature incubator is adjusted to 6 ℃ for continuous culture (the illumination intensity is unchanged), namely, the temperature is set to 2 ℃ (0:00-7:30), 5 ℃ (7:30-10:30), 10 ℃ (10:30-14:30), 5 ℃ (14:30-19:30) and 2 ℃ (19:30-24:00); meanwhile, spraying treatment is carried out on the internal leaves of the Wucai with 1mM MeJA, and the spraying is carried out three times continuously, and 300 mu l of spraying is sprayed each time, so that the leaves are preferably wet.
Example 2
The method for promoting the quick yellowing of the black vegetables in the embodiment comprises the following steps of:
(1) The black vegetable seeds are sown in containers and placed in the following environments for cultivation: day temperature 26 ℃, night temperature 9 ℃ and luminous flux density 500 mu mol.m -2 ·s -1 The relative humidity was 70%.
(2) After germination of the seeds and 4 true leaves, transplanting the seedlings to nutrient soil: vermiculite = 3:1 in plastic pots.
(3) When the black vegetable plant reaches the plant period, the black vegetable plant is placed in an ultralow temperature incubator for low-temperature pretreatment, the temperature is set to 8 ℃ (0:00-7:30), 11 ℃ (7:30-10:30), 16 ℃ (10:30-14:30), 11 ℃ (14:30-19:30), 8 ℃ (19:30-24:00), and the illumination intensity is 300 mu mol m -2 s -1 。
(4) After 2 days of low temperature pretreatment, the temperature of the ultra-low temperature incubator is adjusted to 6 ℃ for continuous culture (the illumination intensity is unchanged), namely, the temperature is set to 2 ℃ (0:00-7:30), 5 ℃ (7:30-10:30), 10 ℃ (10:30-14:30), 5 ℃ (14:30-19:30) and 2 ℃ (19:30-24:00); simultaneously, spraying treatment is carried out on the internal leaves of the Wucai with 1mM MeJA, and three times of spraying are carried out continuously, wherein each time, 350 mul of spraying is carried out, and the wetting of the leaves is preferable.
Example 3
The method for promoting the quick yellowing of the black vegetables in the embodiment comprises the following steps of:
(1) The black vegetable seeds are sown in containers and placed in the following environments for cultivation: day temperature 25 ℃, night temperature 18 ℃, and luminous flux density 400 mu mol.m -2 ·s -1 The relative humidity was 65%.
(2) After germination of the seeds and 4 true leaves, transplanting the seedlings to nutrient soil: vermiculite = 3:1 in plastic pots.
(3) When the black vegetable plant reaches the plant period, the black vegetable plant is placed in an ultralow temperature incubator for low-temperature pretreatment, the temperature is set to 8 ℃ (0:00-7:30), 11 ℃ (7:30-10:30), 16 ℃ (10:30-14:30), 11 ℃ (14:30-19:30), 8 ℃ (19:30-24:00), and the illumination intensity is 300 mu mol m -2 s -1 。
(4) After 2 days of low temperature pretreatment, the temperature of the ultra-low temperature incubator is adjusted to 6 ℃ for continuous culture (the illumination intensity is unchanged), namely, the temperature is set to 2 ℃ (0:00-7:30), 5 ℃ (7:30-10:30), 10 ℃ (10:30-14:30), 5 ℃ (14:30-19:30) and 2 ℃ (19:30-24:00); simultaneously, spraying treatment is carried out on the internal leaves of the Wucai with 1mM MeJA, and the spraying is carried out three times continuously, and 330 mu l of spraying is sprayed each time, so that the leaves are preferably wetted.
Experimental example:
at present, studies have found that the main cause of broccoli yellowing is chlorophyll degradation. By treating broccoli with exogenous methyl jasmonate (MeJA) and diethyl Dithiocarbamate (DIECA), an inhibitor of methyl jasmonate, the endogenous jasmonate content was found to proliferate with the yellowing process. MeJA treatment increased endogenous jasmonic acid and DIECA treatment decreased endogenous jasmonic acid content. Therefore, the present invention attempted to investigate whether it could affect the turning of black vegetable leaves by treating yellow black at low and low temperatures with exogenous methyl jasmonate (MeJA) and diethyl Dithiocarbamate (DIECA), respectively.
Experimental example 1
The experimental example is used for verifying the relation between endogenous jasmonic acid and black vegetable turning yellow.
First, endogenous JA content was measured by targeted metabolism LC-MS/MS technique under low temperature treatment (natural temperature in winter) using 5 kinds of black vegetable varieties (W18-2, WS19-7, W12-7-2, W16-16, W16-7) with different degrees of yellowing as samples (shown in FIG. 1), and the results are shown in Table 1. As can be seen from table 1: the total JA content of the green-yellow variety gradually increases, and the total JA content of the yellow variety with the highest yellowing degree is increased by 93.52 percent compared with the total JA content of the full green variety. The levels of JA-Ile, JA-Val and MeJA were also increased by 76.09%, 47.49% and 51.51%, respectively, indicating that the degree of leaf color yellowing was positively correlated with the endogenous JA levels.
TABLE 1 endogenous JA content of 5 Wucai varieties with different degrees of yellowing
To further verify the change in jasmonic acid content during the yellow turning process of wu-ban, then, the endogenous JA content was measured using targeted metabolism LC-MS/MS technique with leaves of wu-ban variety W7-2 at 5 periods (13, 21, 29, 37, 45 d) before and after low temperature as samples, and the results are shown in table 2. As can be seen from table 2: for the same variety, the blades gradually yellow as the temperature gradually decreases. After 45d of low temperature domestication, the endogenous levels of leaf blades of total JA, JA-Ile, JA-Val and MeJA increased by 135.96%, 132.77%, 157.85% and 152.30%, respectively, indicating a significant increase in the endogenous levels of JA induced at low temperatures.
TABLE 2 endogenous JA content of Wucai W7-2 at various periods
In summary, W7-2 turned yellow within She Hui after undergoing low temperature induction, with a concomitant increase in endogenous jasmonic acid content. Thus, there is a positive correlation between "endogenous jasmonic acid" and the yellow turning of wu.
Experimental example 2
The experimental example is used for researching the change of the yellow of the internal leaves of the Wucai when the Wucai is treated by exogenous methyl jasmonate (MeJA) and an inhibitor of methyl jasmonate (DIECA) respectively, so as to determine the exact relation between the MeJA and the yellow of the Wucai, and screening out the optimal MeJA action concentration (the inhibitor is used as a control).
To determine the concentrations of exogenous methyl jasmonate and inhibitor, the following concentrations were first formulated:
MeJA：0mM、0.1mM、0.2mM、0.5mM、1mM、2mM；
DIECA：0mM、0.1mM、1mM、5mM、10mM。
step one, when W7-2 begins to turn yellow after low-temperature domestication, dividing the W7-2 into three groups: one group was sprayed with distilled water as a control; spraying Wu Cai inner leaves with MeJA with different concentrations in two groups; three groups were sprayed with different concentrations of DIECA on Wu Cai inner leaves. All treatments were independently repeated three times and in all treatments, the wu-sedge was sprayed three times (300 to 350 μl each) with leaf wetting being preferred.
Step two, chlorophyll content determination
The content of photosynthetic pigments (Chl a, chl b) was measured by the Arnon method: fresh leaves were ground to a powder, then 2.5ml of 80% acetone and a small amount of calcium carbonate and quartz sand were added and ground to a homogenate. 3ml of acetone was added thereto and the mixture was left to stand in the dark for 3 to 5 seconds. A piece of filter paper was placed in a funnel, moistened with ethanol, the extract was poured into the funnel along a glass rod and filtered into a 25ml volumetric flask. Sucking 95% ethanol with a dropper, cleaning the mortar, filtering the cleaning solution into a volumetric flask, eluting pigment with 95% ethanol along the periphery of the filter paper, and finally fixing the volume to scale with 95% ethanol after the filter paper and residues are completely whitened. Absorbance at 646 and 663nm wavelengths was measured using a TU1950 ultraviolet visible spectrophotometer (PER-SEE, beijing, china). Wherein, the content of Chl a (mg/g) and Chl b (mg/g) is calculated as follows:
CChl a mg/L＝12.21×A663-2.81×A646；
CChl b mg/L＝20.13×A646-5.03×A663；
Chl a mg/g＝CChl a mg/L×V(L)/Mfresh；
Chl b mg/g＝CChl b mg/L×V(L)/Mfresh。
step three, determination of endogenous jasmonic acid content
Determination of endogenous jasmonic acid content the jasmonic acid content was determined by an enzyme-linked immunosorbent assay. Extraction of JA and ELISA detection using polyclonal anti-JA antibodies employed the following method: 5mL of 80% aqueous methanol was added to 1.0g of the Wucai sample, the mixture was uniformly ground in the presence of liquid nitrogen, incubated overnight at 4℃and then centrifuged at 12000 Xg for 15min at 4℃and the filtrate was discarded and the supernatant containing JA was collected. The precipitate was then extracted twice with 80% methanol. All supernatants were combined, dried under vacuum, and re-dissolved in 1.5mL of extract for ELISA. Plant jasmonate enzyme-linked immunosorbent assay kit (MLBIO, shanghai enzyme-linked biotechnology limited, shanghai, china) was used. The jasmonic acid standard curve was used to calculate the jasmonic acid content based on the absorbance of the standard substance at 450nm according to the manufacturer's instructions. Step four, measuring color values: leaf colorimetric values were measured on the upper surface of the leaf every 3 days after planting using a colorimeter (DS-700D), 10 plants were measured, and the measurement was repeated 3 times at the same leaf positions. And (5) performing color data analysis by adopting a CIELAB color coordinate system. Wherein, the color difference value has three representative color values: l, a, b. L represents brightness (black and white), a represents red-green, and b represents yellow-blue. In addition, there are two chromatic aberrations: value H (Hue angle) and value C (Chroma).
The results are shown in tables 3 and 4.
TABLE 3 influence of different concentrations of MeJA on chlorophyll, JA content, a and b of W7-2 leaves
TABLE 4 influence of different DIECA concentrations on chlorophyll, JA content, a and b of W7-2 leaves
From the above, it is possible to: when compared to the fresh water treatment, the endogenous jasmonic acid content and the leaf color parameter b values (yellow and blue) increased by 21.0% and 47.5%, respectively, and the total chlorophyll content and the leaf color parameter a values (red and green) decreased by 18.8% and 57.7%, respectively, at a MeJA concentration of 1 mM. However, when MeJA concentration was greater than 1mM, the endogenous jasmonic acid content and leaf color parameter b values decreased by 9.5% and 15.8%, respectively, and the total chlorophyll content and leaf color parameter a values increased by 12.8% and 13.1%, respectively, as compared to treatment with 1mMMeJA (table 3). As a result, it was revealed that the effect of promoting the yellowing of Wucai increased with an increase in the MeJA concentration, and that the degree of promoting the yellowing of Wucai was maximized when the MeJA concentration was 1mM, but the inhibition was started when the MeJA concentration was more than 1mM, so that the effect of promoting the yellowing of Wucai was most suitable when the MeJA concentration was 1 mM.
When the DIECA concentration was 1mM, the endogenous jasmonic acid content and the leaf color parameter b values decreased by 9.9% and 7.3%, respectively, and the total chlorophyll content and the leaf color parameter a values increased by 8.33% and 22.7%, respectively, compared to the fresh water treatment. But when the DIECA concentration was greater than 1mM, the endogenous jasmonic acid content and leaf color parameter b values began to rise and the total chlorophyll content and leaf color parameter a values began to fall compared to treatment with 1mMMeJA (table 4). The results show that DIECA can inhibit the yellowing of wu dish, the inhibition being maximal at a DIECA concentration of 1mM, but the inhibition starting to decrease above 1mM, so DIECA concentration of 1mM is the most suitable concentration to inhibit the yellowing of wu dish.
In conclusion, there is a clear and definite relationship between the yellow turning of the internal leaves of Wucai and MeJA, and the concentration of MeJA is 1mM, which is the optimum concentration for promoting the yellow turning of Wucai.
Experimental example 3
The low-temperature pretreatment is needed before the low-temperature treatment of the black vegetable, and different temperature gradients can be helpful for the black vegetable to adapt to the low-temperature environment better and turn yellow faster.
The experimental example is used for researching the influence of different low-temperature pretreatment temperature gradients on the yellowing of the Wucai.
Placing W7-2 in an ultralow temperature incubator, setting 3 different temperature gradients in the time periods of 0:00-7:30, 7:30-10:30, 10:30-14:30, 14:30-19:30 and 19:30-24:00, and measuring the leaf length, leaf width, plant height, development degree, chlorophyll content and endogenous jasmonic acid content of the fourth leaf under the different temperature gradients after two days of treatment so as to screen the temperature gradient more suitable for low-temperature pretreatment of W7-2.
Temperature gradient 1:6 ℃ (0:00-7:30), 9 ℃ (7:30-10:30), 14 ℃ (10:30-14:30), 9 ℃ (14:30-19:30), 6 ℃ (19:30-24:00);
temperature gradient 2:8 ℃ (0:00-7:30), 11 ℃ (7:30-10:30), 16 ℃ (10:30-14:30), 11 ℃ (14:30-19:30), 8 ℃ (19:30-24:00);
temperature gradient 3:10 ℃ (0:00-7:30), 13 ℃ (7:30-10:30), 18 ℃ (10:30-14:30), 13 ℃ (14:30-19:30), 10 ℃ (19:30-24:00).
The results are shown in Table 5.
TABLE 5 growth conditions of W7-2 at different temperature gradients
As can be seen from table 5: under the temperature gradient 2, the chlorophyll content is increased by 15.2% compared with 1, the endogenous jasmonic acid content is reduced by 3.4%, the chlorophyll content is reduced by 13.1% compared with 3, and the endogenous jasmonic acid content is increased by 6.5%. The leaf length, leaf width, plant height and development of W7-2 under temperature gradients 2 and 3 are all significantly higher than those of temperature gradient 1.
The results show that the lower the temperature is, the formation of endogenous jasmonic acid is promoted, and the growth and chlorophyll content of the Wucai are inhibited to a certain extent. Therefore, under the condition of the temperature gradient 2, the growth condition of the W7-2 is ensured to be good, and the yellowing is promoted to a certain extent.
In summary, the temperature for the low temperature pretreatment of W7-2 was set to 8 ℃ (0:00-7:30), 11 ℃ (7:30-10:30), 16 ℃ (10:30-14:30), 11 ℃ (14:30-19:30), 8 ℃ (19:30-24:00).
Experimental example 4
After the W7-2 is subjected to low-temperature pretreatment, the W7-2 is subjected to low-temperature treatment, and the experimental example is used for researching the influence of different low-temperature treatment temperature gradients after the low-temperature pretreatment on the yellowing of the Wucai.
Based on experimental example 3, 8 ℃ (0:00-7:30), 11 ℃ (7:30-10:30), 16 ℃ (10:30-14:30), 11 ℃ (14:30-19:30) and 8 ℃ (19:30-24:00) were selected as the optimal low temperature pretreatment mode, then three different 'low temperature treatment' temperature gradients were set, and the fourth leaf length, leaf width, plant height, development, chlorophyll content and endogenous jasmonic acid content at different temperature gradients were measured after two days of treatment to screen the temperature gradient more suitable for low temperature treatment of W7-2.
Temperature gradient 1:4 ℃ (0:00-7:30), 7 ℃ (7:30-10:30), 12 ℃ (10:30-14:30), 7 ℃ (14:30-19:30), 4 ℃ (19:30-24:00);
temperature gradient 2:2 ℃ (0:00-7:30), 5 ℃ (7:30-10:30), 10 ℃ (10:30-14:30), 5 ℃ (14:30-19:30), 2 ℃ (19:30-24:00);
temperature gradient 3:0 ℃ (0:00-7:30), 3 ℃ (7:30-10:30), 8 ℃ (10:30-14:30), 3 ℃ (14:30-19:30), 0 ℃ (19:30-24:00).
The results are shown in Table 6.
TABLE 6 growth conditions of W7-2 at different temperature gradients
As can be seen from table 6: w7-2 had no significant differences in leaf length, leaf width, plant height and development at temperature gradients 1 and 2, but these several indices were significantly lower than 1 and 2 at temperature gradient 3. Under the temperature gradient 2, the chlorophyll content is reduced by 11.1% compared with 1, the endogenous jasmonic acid content is increased by 3.5%, the chlorophyll content is increased by 74.0% compared with 3, and the endogenous jasmonic acid content is reduced by 5.1%. The results show that under the condition of the temperature gradient 3, the blade area is reduced due to excessive temperature down-regulation, chlorophyll is destroyed, the jasmonic acid content is obviously increased, and the growth of W7-2 is not suitable. And under the temperature gradient 2, the yellowing of W7-2 is promoted while the good growth condition of W7-2 is ensured.
In summary, the temperature of the low-temperature pretreatment of W7-2 is set to 8 ℃ (0:00-7:30), 11 ℃ (7:30-10:30), 16 ℃ (10:30-14:30), 11 ℃ (14:30-19:30), 8 ℃ (19:30-24:00), and the temperature of the ultra-low temperature incubator is adjusted to 6 ℃ to be optimal after 2 days of the low-temperature pretreatment.
Experimental example 5
Based on experimental example 4, the experimental example is used for comprehensively verifying the influence of different treatments (clear water, meJA or DIECA treatment) at low temperature and normal temperature on chlorophyll, JA content, a and b of the W7-2 leaf.
1. The adult stage of W7-2 is divided into two parts:
a part of: low temperature pretreatment in an ultra-low temperature incubator at 8deg.C (0:00-7:30), 11deg.C (7:30-10:30), 16deg.C (10:30-14:30), 11deg.C (14:30-19:30), 8deg.C (19:30-24:00), and light intensity of 300 μm -2 s -1 The method comprises the steps of carrying out a first treatment on the surface of the After 2 days of low temperature pretreatment, the temperature of the ultra-low temperature incubator was adjusted down to 6 ℃.
Another part: placing in room temperature growth chamber, setting temperature at 26+ -2deg.C (daytime) and 18+ -2deg.C (nighttime), and illumination intensity of 300 μmol m -2 s -1 。
2. The W7-2 leaves that began to yellow at the beginning of the plant stage and W7-2 leaves that did not yellow at room temperature were treated with 1mM MeJA and 1mM DIECA, and the control was clear water.
The results are shown in tables 7 and 8.
TABLE 7 influence of different treatments at low temperature on chlorophyll, JA content, a and b of W7-2 leaves
TABLE 8 influence of different treatments at normal temperature on chlorophyll, JA content, a and b of W7-2 leaves
The result shows that:
at low temperature, compared with the control, after MeJA treatment, chlorophyll content and a value are respectively reduced by 17% and 63.1%, and JA content and b value are respectively increased by 24.3% and 46.9%; after DIECA treatment, chlorophyll content and a value were increased by 10.6% and 8.4%, respectively.
At normal temperature, compared with untreated water, the inhibitor treated water has no obvious change in the chlorophyll content, JA content and leaf color parameters of W7-2; after MeJA treatment, chlorophyll content and a value were reduced by 12.5% and 43.2%, respectively, and JA content and b value were increased by 17% and 36.5%, respectively. Compared with W7-2 at low temperature, chlorophyll content and a value at normal temperature are respectively increased by 7.7% and 31.7% after MeJA treatment, and JA content and b value are respectively decreased by 7.5% and 9.7%.
The results show that the exogenous MeJA can turn black vegetable into yellow at normal temperature, and can promote the black vegetable to turn yellow rapidly at low temperature, and the black vegetable leaves are gradually greened after DIECA treatment (as shown in figure 2).
Experimental example 6
On the basis of experimental example 5, W7-2 leaves at low temperature were treated with MeJA at a concentration of 1mM, samples were collected every two days, and changes in chlorophyll, JA content, a and b values of W7-2 leaves (with clear water treatment as a control) were measured at different times ( days 0,2, 4, 6, 8, 10).
The results are shown in FIGS. 3, 4, 5 and 6,
As can be seen from fig. 3 to 6: the chlorophyll content and a values were significantly lower than untreated and the JA content and b values were significantly higher than untreated for W7-2 after MeJA treatment from day six, so that over the course of the treatment days, the treated group turned yellow in advance compared to the control group, indicating that MeJA treatment rapidly promoted yellowing of the W7-2 inner leaves.
From the above experimental examples, it is comprehensively known that low temperature and exogenous methyl jasmonate are a way for rapidly promoting the yellow turning of the Wucai.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, and alternatives falling within the spirit and principles of the invention.
Claims (5)
1. The method for promoting the quick yellowing of the black vegetables is characterized by comprising the following specific steps of:
(1) Sowing the black vegetable seeds in a container, and culturing the seeds;
(2) Transplanting after the seeds germinate and have 4 true leaves;
(3) When the black vegetable plants reach the plant period, placing the black vegetable plants in an ultralow temperature incubator for low-temperature pretreatment, wherein the pretreatment temperature is as follows: 0:00-7:30,8 ℃;7:30-10:30, 11 ℃;10:30-14:30, 16 ℃;14:30-19:30, 11 ℃;19:30-24:00,8 ℃;
(4) After 2 days of low-temperature pretreatment, the temperature of the ultralow-temperature incubator is adjusted to 6 ℃ for continuous culture, namely, the temperature is 0:00-7:30 and 2 ℃;7:30-10:30,5 ℃;10:30-14:30, 10 ℃;14:30-19:30,5 ℃;19:30-24:00,2 ℃; simultaneously, spraying treatment is carried out on the internal leaves of the Wucai with 1mM MeJA, and the spraying treatment is carried out three times continuously, wherein 300-350 mu l of the internal leaves of the Wucai are sprayed each time.
2. The method for promoting rapid yellowing according to claim 1, wherein in the step (1), the culture environment of the black vegetable seeds is: day temperature 25+ -1 deg.C, night temperature 18+ -1 deg.C, light quantum flux density 300-500 mu mol.m -2 ·s -1 The relative humidity is 60-70%.
3. The method for promoting quick yellowing according to claim 1, wherein in the step (2), transplanting the black vegetable seedlings to nutrient soil: vermiculite = 3:1 in plastic pots.
4. The method for promoting rapid turning yellow of wu-cai according to claim 1, wherein the illumination intensity is set to 300 μmolm during the low-temperature pretreatment in the step (3) -2 s -1 。
5. The method for promoting rapid yellowing according to claim 1, wherein in the step (4), the illumination intensity is set to 300 μmolm when culturing at a temperature of 6 ℃ -2 s -1 。
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