CN107787973B - Application of rhamnolipid in relieving plant salt stress - Google Patents
Application of rhamnolipid in relieving plant salt stress Download PDFInfo
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Images
Classifications
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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
-
- 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
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/30—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests characterised by the surfactants
Abstract
The invention researches the changes of the relative growth rate, the relative water content, the relative conductivity and the Malondialdehyde (MDA) content of zoysia japonica in the salt stress process and after the rhamnolipid is used to prove the effect of the rhamnolipid in relieving the salt stress of zoysia japonica, and finds that the rhamnolipid can obviously improve the relative water content and the relative growth rate of the zoysia japonica in the salt stress process, reduce the relative conductivity and the malondialdehyde content of the zoysia japonica, thereby relieving the harm of the salt stress to the zoysia japonica, and providing a new method for relieving the salt stress of the zoysia japonica in the saline-alkali soil; the optimal concentration of the rhamnolipid solution is 0.5 g/L.
Description
Technical Field
The invention relates to a new application of rhamnolipid, in particular to an application in relieving zoysia salt stress.
Background
Zoysia japonica often grows in warm and humid coastal areas, has strict requirements on illumination, has certain shadiness, drought resistance, salt and alkali resistance and strong pest resistance, and is commonly used for ornamental lawns, sports lawns and slope protection lawns. Zoysia japonica grows in open environment like other plants and is often influenced by adverse environment, and severe environments such as high temperature, low temperature, salinization and drought can inhibit the growth of the zoysia japonica, so that the lawn quality is reduced.
The area of the saline-alkali land is about 9.6 multiplied by 108hm2And 3.47 multiplied by 10 for salinized land in China7hm2This corresponds to 1/3 of the cultivated land area. The plant growth can be inhibited when the soil contains too much salt, and the harm of the soil salinization to the plants is reflected in that: na in cells+The excessive accumulation can break the ion balance in cells, inhibit the normal metabolism in plant cells, reduce the photosynthesis capability of plants, and lead the growth of the plants to lack nutrients and then be inhibited and even die finally; the salinity of the saline-alkali soil enables the osmotic pressure of the soil to be higher than that of a plant root system, so that the water absorption of the plant is difficult, and the plant is dehydrated and dried up; the salinization of the land can destroy the balance of generation and elimination of active oxygen in plants, cause the accumulation of a large amount of free radicals, generate membrane lipid peroxidation, destroy cell membranes and enable organic matters of cells to permeate outwards, thereby inactivating the cells. Research finds that the salt resistance of plants can be improved by using exogenous trehalose through increasing the active oxygen scavenging capacity, relieving plasma membrane damage and maintaining the stable state of cytoplasmic ions. The rare earth solution is used for seed dressing or field spraying to enhance the effect of plant cell membranes on electrolytesThe control ability of infiltration stabilizes the metabolic process in plant cells and improves the salt tolerance of plants. Spraying H in the salt stress process2O2Can increase the content of osmoregulation substances such as soluble protein, soluble sugar and proline in plant, increase the activity of antioxidase such as SOD, CAT, POD and APX, increase the content of antioxidant GSH, and reduce O2 -And H2O2Accumulating, reducing membrane lipid oxidation injury and plant growth inhibition degree, thereby enhancing plant salt tolerance.
Rhamnolipids are a very important class of biosurfactants produced by pseudomonas aeruginosa. The biosurfactant is a metabolic substance with certain biochemical activity, is generated in vitro or on cell membranes by microorganisms such as bacteria, fungi, yeasts and the like, and can also be obtained by physiological and biochemical reactions of animals and plants. The biosurfactant has the characteristics of easy degradation, low toxicity, wide sources, high biocompatibility, multiple functional groups, strong adaptability and the like, can improve the hydrophobic capacity of the cell surface, and increases the direct physical action between a slightly soluble substrate and plant cells. Researches show that the rhamnolipid not only can change the surface property of cells and increase the hydrophobicity of the cells, but also can obviously improve the solubility of hydrocarbon compounds. In addition, the biological surfactant can influence the antibacterial capacity of microorganisms and the regulation and control capacity of microorganisms on the adsorption and desorption of plant cell membranes, and has great influence on heavy metal combination, the formation of biological membranes and the like. The research on the prevention and treatment effect of rhamnolipid on watermelon fusarium wilt proves that the rhamnolipid can prevent and treat watermelon seedling fusarium wilt by inducing plant resistance in a proper environment, and the research shows that the irrigation treatment of the rhamnolipid can improve the activities of chitinase and glucanase in plants. However, the effect of the plant salt stress relieving agent on the aspect of relieving plant salt stress is not reported so far.
Disclosure of Invention
The invention researches the changes of the relative growth rate, the relative water content, the relative conductivity and the Malondialdehyde (MDA) content of zoysia japonica in the salt stress process and after the rhamnolipid is used to prove the effect of the rhamnolipid in relieving the salt stress of zoysia japonica, and finds that the rhamnolipid can obviously improve the relative water content and the relative growth rate of the zoysia japonica in the salt stress process, reduce the relative conductivity and the malondialdehyde content of plants, thereby relieving the harm of the salt stress to the plants and providing a new method for relieving the salt stress of the zoysia japonica in the saline-alkali soil.
Specifically, the rhamnolipid can be used for relieving the harm of salt stress on zoysia japonica, and the application can be realized by irrigating a rhamnolipid solution, preferably, the concentration of the rhamnolipid solution is 0.5g/L, and detection shows that the rhamnolipid can obviously improve the relative water content and the relative growth rate of zoysia japonica leaves in the salt stress process and reduce the relative conductivity and the malondialdehyde content of the zoysia japonica.
Drawings
FIG. 1 is a graph showing the change in the relative growth rate of zoysia japonica during salt stress in an example of the present invention;
FIG. 2 is a graph showing the change in relative conductivity of zoysia japonica during salt stress in an example of the present invention;
FIG. 3 is a graph showing the change in water content of zoysia japonica leaves during salt stress in an embodiment of the present invention;
FIG. 4 is a graph showing the change in malondialdehyde content of zoysia japonica during salt stress in examples of the present invention;
FIG. 5 is a graph showing the change in the relative growth rate of zoysia japonica after administration of rhamnolipids in an example of the present invention;
FIG. 6 is a graph of the change in relative conductivity of zoysia japonica after administration of rhamnolipids in an example of the present invention;
FIG. 7 is a graph showing the change in water content of zoysia japonica leaves after administration of rhamnolipids in an example of the present invention;
FIG. 8 is a graph of the change in malondialdehyde content of zoysia following administration of rhamnolipids in an example of the invention;
in the figure: the difference between different lower case letters is significant (P < 0.05).
Detailed Description
In order that the objects and advantages of the invention will be more clearly understood, the invention is further described in detail below with reference to examples. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The applicant has obtained through a large number of experiments: when the concentration of the rhamnolipid is 0.5g/L, the effect of relieving the zoysia salt stress is the best, so the rhamnolipid with the concentration of 0.5g/L is selected as an experimental reagent in the following examples.
Examples
Examples zoysia matrella and zoysia sinensis were used as materials, and turf pieces (diameter of 12.00cm) of both zoysia japonica were planted in pots (height of 14.00cm, pot mouth diameter of 13.00cm) which were not permeable to water, 6 pots each were planted, of which 3 pots were used for salt treatment and 3 pots were used for control. The substrate for salt stress treatment is raw soil from the beach of the salt city, and is prepared by mixing to keep the salt concentration (conductivity) at about 0.5%, placing the culture substrate into a culture pot, compacting, planting turf blocks on the culture substrate, and culturing with soil free of salt as control. The cells were incubated at room temperature (incubation temperature 20. + -. 5 ℃ C.) and poured with 600mL of water each week. After cultivation, each pot material was trimmed to 2.00cm from the pot mouth, and the physiological index was measured by taking the leaf at 4d, 8d, 12d, 16d and 20d after cultivation. Then treating 100mL of rhamnolipid with concentration of 0.5g/L per pot, and taking out leaves again at 4d, 8d, 12d, 16d and 20d after application to determine physiological index.
The method for measuring the physiological indexes comprises the following steps:
1. relative growth rate
The vertical growth height of each zoysia japonica was measured, and the relative growth rate was obtained by dividing the vertical growth height of each group treatment by the growth height of the control group.
2. Relative water content of leaves
Cutting a plurality of zoysia matrella and zoysia sinensis leaves, quickly putting the zoysia matrella and zoysia sinensis leaves into an aluminum box with known weight, and weighing the fresh weight of zoysia japonica leaf tissues. Weighing fresh weight of plant tissue, soaking zoysia japonica leaves in distilled water for 5-9h until the water content of the zoysia japonica leaves is close to saturation state, then putting the zoysia japonica leaves in a drying oven at 105 ℃ for deactivating enzymes for 15min, then putting the zoysia japonica leaves in the drying oven at 80 ℃ for drying to constant weight, and weighing dry weight of the tissue. The relative water content of the leaves was calculated using the following formula:
relative water content ═ plant tissue fresh weight-plant tissue dry weight)/(plant tissue saturated fresh weight-plant tissue dry weight)
3. Relative conductivity measuring method
Taking zoysia japonica leaves at the same position, cutting the zoysia japonica leaves into 0.6cm small sections by using scissors, quickly flushing the zoysia japonica leaves twice by using double distilled water, wrapping the zoysia japonica leaves by using absorbent paper to absorb water, weighing 0.3g of the processed zoysia japonica leaves, putting the zoysia japonica leaves into a coarse test tube, adding 6mL of the double distilled water, carrying out vacuum infiltration, air exhaust and air exhaust for 3 times, enabling water to be tightly contacted with the leaves to enable electrolyte to be easily seeped out, taking out the zoysia japonica leaves after 30min, standing the zoysia japonica leaves at room temperature for 2h, measuring a conductivity value by using a conductivity meter, taking the conductivity value as an original conductivity value, heating the zoysia japonica leaves at 100 ℃ for 5min to kill the zoysia japonica leaves, cooling. The relative conductivity of the blades was calculated using the following formula.
Relative conductivity (%) < original conductance/total conductance x 100%
4. Malondialdehyde (MDA) content determination method
Weighing 0.5g of chopped zoysia japonica leaves, adding 2mL of 10% chloroacetic acid (TCA) and a small amount of quartz sand, grinding until homogenate is obtained, adding a proper amount of TCA for further grinding, transferring the homogenate into a 5mL centrifuge tube, centrifuging at 4000rpm for 10min, and taking supernatant as a sample extracting solution. 2mL of the centrifuged supernatant (2 mL of distilled water added to the blank) was taken, 2mL of a 0.6% TBA (thiobarbituric acid) solution was added, and the mixture was reacted in a boiling water bath for 15min, rapidly cooled and centrifuged. The supernatant was taken to determine the optical density at wavelengths of 532nm, 600nm and 450 nm.
MDA content (μmol/gfr) ═ { MDA concentration (μmol/L) × extract volume (mL) }/fresh weight of plant tissue (g)
Results and analysis
Effect of salt stress on the relative growth rates of two species of zoysia
The change in relative growth rate of the two species of zoysia during salt stress is shown in figure 1. As can be seen from FIG. 1, at the initial stage of salt stress, i.e., from the 4 th d to the 8 th d, the relative growth rates of both zoysia matrella and zoysia huashanensis increased by 4%, and the relative growth rates of zoysia matrella increased by 3%, indicating that the salt stress for a short period of time contributes to the growth of zoysia japonica, but there was no significant difference (P > 0.05) between the two zoysia japonica. After 8d, the relative growth rate of the two zoysia japonica shows a gradual reduction trend along with the prolonging of the salt stress time. The relative growth rate change ranges of zoysia matrella and zoysia sinensis are respectively 99% -93% and 97% -89%; at 12d, 16d and 20d of salt stress, the relative growth rate of zoysia sinensis is significantly smaller than that of zoysia japonica of ditch leaves (P < 0.05), which indicates that the salt stress significantly affects the relative growth rates of the two zoysia japonica.
Effect of salt stress on relative conductivity of two zoysia japonica leaves
The relative conductivities of the two zoysia japonica during salt stress are shown in fig. 2, and as can be seen from fig. 2, the relative conductivities of the controls do not change much and have no significant difference (P > 0.05). The relative conductivity of the leaves of the two zoysia japonica shows a gradual rising trend in the salt stress process, and the variation ranges of the relative conductivity of the leaves of the zoysia japonica and the zoysia sinensis are 8.9% -33.26% and 9.21% -40.85%, respectively. At 4d of salt stress, the relative conductivity of zoysia japonica and zoysia sinensis is increased, but has no significant difference with the control, and has no significant difference between treatments (P is more than 0.05). At 8d, the relative conductivity of zoysia matrella was not significantly different from the control (P > 0.05), but the relative conductivity of zoysia sinensis was significantly higher than that of zoysia matrella (P < 0.05). At 12d, 16d and 20d of salt stress, the relative conductivity of the two zoysia japonica is not only significantly higher than that of the control, but also that of the zoysia sinensis is significantly higher than that of zoysia japonica of furrows (P < 0.05).
Effect of salt stress on Water content of two leaves of zoysia japonica
The water content of the leaves of the two zoysia japonica during the salt stress process is shown in figure 3, and the water content of the control leaves is not changed greatly and has no significant difference (P is more than 0.05) from figure 3. In the salt stress process, the relative water content of the leaves of the two zoysia japonica gradually decreases, and the relative water content of the leaves of the zoysia japonica and the zoysia sinensis is 78.45-55.74% and 76.38-42.62% respectively. The relative water content of zoysia sinensis starting from 4d is significantly lower than that of the control (P < 0.05), while the relative water content of zoysia japonica is significantly lower than that of the control (P < 0.05) starting from 8d, and the relative water content of zoysia japonica treated at 16d and 20d is significantly greater than that of zoysia sinensis (P < 0.05).
Effect of salt stress on the malondialdehyde content of two species of zoysia
The change of malondialdehyde content of the two zoysia japonica during salt stress is shown in figure 4, and as can be seen from figure 4, the change of malondialdehyde content of the control group is not large, and the two have no significant difference (P is more than 0.05). The malondialdehyde content of two zoysia japonica in the salt stress process is gradually increased, and the malondialdehyde content of the zoysia japonica furrows and zoysia sinensis leaves is respectively 40-190 [ mu ] moL/g and 45-240 [ mu ] moL/g. At the 4 th d and 8 th d of salt stress, the contents of the zoysia matsutake and zoysia media hamiana malondialdehyde have no significant difference with the control, and the treatment has no significant difference (P is more than 0.05). Starting from 12d, the content of zoysia matrella and zoysia media malondialdehyde was significantly higher than the control (P < 0.05), and the content of zoysia media malondialdehyde was significantly higher than that of zoysia matrella (P < 0.05).
Changes in the relative growth rates of two species of zoysia after rhamnolipid administration
The change of the relative growth rate of the two zoysia japonica leaves after the rhamnolipid administration is shown in fig. 5, and it can be seen from fig. 5 that the relative growth rate of the two zoysia japonica leaves after the rhamnolipid administration is continuously increased, and the change ranges of the relative growth rate of the leaves of zoysia japonica and zoysia sinensis are 94% -98% and 90% -97%, respectively. When rhamnolipid 20d was administered, the relative growth rate of zoysia japonica was increased by 5% compared to salt-stressed 20d and the relative growth rate of zoysia sinensis was increased by 8% compared to salt-stressed 20 d. The relative growth rate of zoysia matrella was significantly greater than that of zoysia sinensis (P < 0.05) when applied at 4d, 8d, 12d and 16d of rhamnolipids. There was no significant difference in the relative growth rates of zoysia matrella and zoysia sinensis leaves at 20d (P > 0.05).
Changes in the relative conductivity of the two zoysia leaves following rhamnolipid administration
The change of the relative conductivity of the two zoysia leaves after the rhamnolipid application is shown in figure 6, and it can be seen from figure 6 that the relative conductivity of the two zoysia leaves after the rhamnolipid application shows a gradual decrease trend, and the change ranges of the relative conductivity of the leaves of zoysia japonica and zoysia sinensis are 33.26% -12.54% and 40.85% -15.42%, respectively. When rhamnolipid 20d was administered, the relative conductivity of zoysia japonica leaves was reduced by 20.72% compared to salt stress 20d and the relative conductivity of zoysia sinensis leaves was reduced by 25.43% compared to salt stress 20 d. The relative conductivity difference between the two zoysia japonica was significant (P < 0.05) at 4d, 8d, 12d and 16d of rhamnolipid application, but there was no significant difference in the relative conductivity of the leaves of zoysia japonica and zoysia sinensis at 20d (P > 0.05).
Changes in moisture content of two zoysia japonica leaves after rhamnolipid administration
The change of the relative water content of the two zoysia leaves after the rhamnolipid is applied is shown in figure 7, and it can be seen from figure 7 that the relative water content of the two zoysia leaves after the rhamnolipid is applied is in a gradually rising trend, and the relative water content of the zoysia japonica leaves and the zoysia sinensis leaves is respectively 55.74% -73.84% and 42.62% -70.56%. When rhamnolipid 20d was administered, the relative water content of zoysia japonica leaves was increased by 18.1% compared to salt stress 20d and the relative water content of zoysia sinensis was increased by 27.94% compared to salt stress 20 d. The relative water content difference between the two zoysia japonica parts was significant (P < 0.05) at 4d, 8d and 12d on rhamnolipids, but not significant (P > 0.05) on leaves of zoysia japonica and zoysia sinensis at 16d and 20 d.
Change in malondialdehyde content of two zoysia after rhamnolipid administration
The change of malondialdehyde content in leaves of two zoysia japonica after rhamnolipid administration is shown in FIG. 8. it can be seen from FIG. 8 that the malondialdehyde content in leaves of two zoysia japonica after rhamnolipid administration is continuously reduced, and the change ranges of malondialdehyde content in leaves of zoysia japonica and zoysia sinensis are 190 μmoL/g-86 μmoL/g and 240 μmoL/g-144 μmoL/g, respectively. When rhamnolipid 20d was applied, the malondialdehyde content of zoysia japonica leaves was 54.7% lower than that of salt-stressed 20d, and the malondialdehyde content of zoysia sinensis leaves was 52.5% lower than that of salt-stressed 20 d. The malondialdehyde content of zoysia japonica was always significantly lower after rhamnolipid administration than that of zoysia sinensis leaves (P < 0.05).
Therefore, after the rhamnolipid is applied, the relative water content of the two zoysia japonica is obviously increased, and the rhamnolipid can increase the osmotic potential of the zoysia japonica root system cells by promoting the synthesis of macromolecules such as protein, sugar and the like in the zoysia japonica root system cells, so that the zoysia japonica can absorb water from soil and increase the transportation to stem leaves. The relative growth rate of the two zoysia japonica after the rhamnolipid is applied shows an increasing trend, which is probably that the rhamnolipid can reduce the absorption of sodium ions in cells, increase the absorption of potassium ions in soil and promote photosynthesis, thereby relieving the inhibition effect of salt stress on the growth of the zoysia japonica. After the rhamnolipid is used, the water content of the leaves is increased, so that the generation and the elimination of active oxygen of the leaves tend to be balanced, the peroxidation degree of membrane substances is reduced, and the MDA content is reduced.
In conclusion, the rhamnolipid is shown to obviously improve the relative water content and the relative growth rate of two zoysia japonica leaves in the salt stress process, reduce the relative conductivity and the malondialdehyde content of the two zoysia japonica leaves and relieve the harm of the salt stress to the zoysia japonica.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that those skilled in the art can make various improvements and modifications without departing from the principle of the present invention, and these improvements and modifications should also be construed as the protection scope of the present invention.
Claims (1)
1. The application of the rhamnolipid in relieving plant salt stress is characterized in that the damage of the salt stress to zoysia japonica can be relieved by pouring a rhamnolipid solution;
the concentration of the rhamnolipid solution is 0.5 g/L;
the rhamnolipid can obviously improve the relative water content and the relative growth rate of zoysia japonica leaves in the salt stress process and reduce the relative conductivity and the malondialdehyde content of the zoysia japonica.
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