CN116349676B - Application of trehalose in alleviating heavy metal stress on okra seedlings - Google Patents
Application of trehalose in alleviating heavy metal stress on okra seedlings Download PDFInfo
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Classifications
-
- 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
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
-
- 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
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P21/00—Plant growth regulators
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Botany (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Pest Control & Pesticides (AREA)
- Biodiversity & Conservation Biology (AREA)
- Ecology (AREA)
- Forests & Forestry (AREA)
- Zoology (AREA)
- Plant Pathology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Chemical & Material Sciences (AREA)
- Agronomy & Crop Science (AREA)
- Health & Medical Sciences (AREA)
- Dentistry (AREA)
- General Health & Medical Sciences (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Cultivation Of Plants (AREA)
Abstract
本发明公开了海藻糖在缓解重金属对秋葵幼苗胁迫作用中的应用，属于缓解植物重金属胁迫技术研究领域。本发明保护海藻糖在缓解秋葵CdCl2胁迫损伤中的应用。本发明以秋葵品种台湾五福为试验材料，研究海藻糖与CdCl2胁迫下秋葵的渗透调节、叶片损伤、光合作用、抗氧化酶系统、细胞膜系统的关系，探讨海藻糖在植物抵御重金属胁迫的作用机理，为其在秋葵的安全生产提供理论基础；本发明将在农业领域具有广阔的应用空间和市场前景。
The invention discloses the application of trehalose in alleviating the stress of heavy metals on okra seedlings, and belongs to the research field of technology for alleviating heavy metal stress of plants. The invention protects the application of trehalose in alleviating the damage of okra under CdCl2 stress. The invention uses the okra variety Taiwan Wufu as the test material, studies the relationship between trehalose and the osmotic regulation, leaf damage, photosynthesis, antioxidant enzyme system, and cell membrane system of okra under CdCl2 stress, explores the mechanism of action of trehalose in plants resisting heavy metal stress, and provides a theoretical basis for its safe production in okra; the invention will have broad application space and market prospects in the agricultural field.
Description
Technical Field
The invention belongs to the technical field of research for relieving heavy metal stress of plants, and particularly relates to application of exogenous trehalose to a stress relieving effect of CdCl 2 of okra seedlings.
Background
Cadmium is a non-essential nutrient element for plants. The low concentration cadmium plays a certain role in promoting plant growth, but the high concentration cadmium has a great toxic effect on plants. With the development of industry, industrial wastewater and pollutants enter soil to pollute the environment, and the problem of cadmium pollution in soil in China is serious. Studies show that excessive cadmium also causes stress on plants, inhibits synthesis of chlorophyll and photosynthesis, causes symptoms of slow growth, short plant, green-removing, yield reduction and the like of the plants under the stress of cadmium, and high-concentration cadmium can destroy active oxygen metabolism balance in the plants, and increase accumulation of active oxygen leads to peroxidation of membrane lipid and reduction of activity of antioxidant enzyme in the plants.
Okra contains abundant proteins, free amino acids, carotenoid, various vitamins, mineral elements such as phosphorus, iron, potassium, calcium and the like, and viscous substances composed of pectin, polysaccharide and the like, has various health care functions, and is welcomed by wide consumers. Research shows that the growth of okra leaves under cadmium stress and the activity of antioxidant enzymes are inhibited, the quality of okra fruits under cadmium stress is reduced, and the accumulation of cadmium in the fruits also affects the health of human beings. Therefore, the alleviation of the damage of cadmium stress to okra is of great importance.
Trehalose (Trehalose, abbreviated as Tre) is a safe and reliable natural sugar, is non-reducing sugar formed by two glucose molecules through 1, 1-glycosidic bonds, is disaccharide with the most stable properties in sugar, has low content in plant bodies, and can stabilize the structure and function of protein by regulating the content of trehalose in plants under adverse conditions. Research shows that trehalose is a penetration regulating substance, not only can protect an antioxidant enzyme system and subcellular structures, but also can mediate penetration regulation and activate ROS clearance, and can relieve damage to plants caused by stress.
Disclosure of Invention
The invention aims to solve the problem that the Cd content of okra seedlings growing under the stress of CdCl 2 is high, and provides application of trehalose in relieving the stress effect of CdCl 2 of okra seedlings.
In order to achieve the purpose, the invention adopts the following technical scheme:
application of trehalose in relieving plant CdCl 2 stress injury or improving plant CdCl 2 stress resistance.
Use of trehalose in at least one of the following (a 1) - (a 33):
(a1) Improving the trehalose content of okra under CdCl 2 stress;
(a2) Preparing a product for increasing the trehalose content of okra under the stress of CdCl 2;
(a3) The proline content of okra under CdCl 2 stress is improved;
(a4) Preparing a product for increasing the proline content of okra under the stress of CdCl 2;
(a5) The chlorophyll a content of okra under CdCl 2 stress is improved;
(a6) Preparing a product for improving chlorophyll a content of okra under CdCl 2 stress;
(a7) The chlorophyll b content of okra under CdCl 2 stress is improved;
(a8) Preparing a product for increasing the chlorophyll b content of okra under the stress of CdCl 2;
(a9) The total chlorophyll content of okra under CdCl 2 stress is improved;
(a10) Preparing a product for improving the total chlorophyll content of okra under the stress of CdCl 2;
(a11) The total carotenoid content of okra under CdCl 2 stress is improved;
(a12) Preparing a product for increasing the total carotenoid content of okra under the stress of CdCl 2;
(a13) The relative water content of okra under CdCl 2 stress is improved;
(a14) Preparing a product for increasing the relative water content of okra under the stress of CdCl 2;
(a15) Improving SOD activity of okra under CdCl 2 stress;
(a16) Preparing a product for improving SOD activity of okra under CdCl 2 stress;
(a17) The GPX activity of okra under CdCl 2 stress is improved;
(a18) Preparing a product for improving GPX activity of okra under CdCl 2 stress;
(a19) Improving CAT activity of okra under CdCl 2 stress;
(a20) Preparing a product for improving CAT activity of okra under CdCl 2 stress;
(a21) Improving GR activity of okra under CdCl 2 stress;
(a22) Preparing a product for improving GR activity of okra under CdCl 2 stress;
(a23) Improving the APX activity of okra under CdCl 2 stress;
(a24) Preparing a product for improving the APX activity of okra under the stress of CdCl 2;
(a25) The POD activity of okra under CdCl 2 stress is improved;
(a26) Preparing a product for improving POD activity of okra under CdCl 2 stress;
(a27) Reducing MDA content of okra under CdCl 2 stress;
(a28) Preparing a product for reducing MDA content of okra under CdCl 2 stress;
(a29) The electrolyte permeability of okra under the stress of CdCl 2 is reduced;
(a30) Preparing a product for reducing the electrolyte permeability of okra under the stress of CdCl 2;
(a31) Reducing the H 2O2 content of okra under CdCl 2 stress;
(a32) Preparing a product for reducing the H 2O2 content of okra under the stress of CdCl 2;
(a33) Improving stress resistance of okra CdCl 2.
In the application, the trehalose with the concentration of 1-100 mu mol/L is adopted to carry out foliar spraying on plants.
A culture method of okra comprises spraying trehalose with concentration of 1-100 mmol/L onto leaf surface of okra. Preferably, the okra is subjected to foliar spraying by adopting the trehalose with the concentration of 1-10 mmol/L, and further preferably, the okra is subjected to foliar spraying by adopting the trehalose with the concentration of 10 mmol/L.
A method for relieving stress injury of okra CdCl 2 adopts trehalose with the concentration of 1-100 mmol/L to carry out foliar spray on the okra. Preferably, the okra is subjected to foliar spraying by adopting the trehalose with the concentration of 1-10 mmol/L, and further preferably, the okra is subjected to foliar spraying by adopting the trehalose with the concentration of 10 mmol/L.
The above plant is any one of the following (c 1) to (c 3):
(c1) Dicotyledonous plants;
(c2) Plants of the family Malvaceae;
(c3) Okra.
The invention also protects the application of the trehalose in the selection and breeding of okra against CdCl 2 stress.
The invention also protects the application of the method in the selection and breeding of the okra resisting CdCl 2 stress.
In any of the above methods, the okra is specifically okra in a seedling stage.
Any of the above okra may be okra of Taiwan five-Fu.
The application of the exogenous trehalose in relieving the stress effect of CdCl 2 on okra seedlings provided by the invention is that the exogenous trehalose sprayed can improve the CdCl 2 stress resistance of okra plants. 0.1 The test result of the stress germination rate of the mmol/L CdCl 2 shows that the germination rate and fresh weight of okra seeds treated by 1 and 10 mmol/L trehalose are obviously higher than those treated by 0 and 100 mmol/L trehalose, and are respectively improved by 51%, 130% and 47% and 118% compared with 0 mmol/L trehalose plants, wherein the okra seedlings treated by 10 mmol/L trehalose have the best effect. 0.1 The test result of the potted plant stressed by mmol/L CdCl 2 shows that the plant treated by exogenous trehalose grows well and the resistance is improved, the fresh weight and the dry weight of the potted plant treated by trehalose are obviously higher than those of the plant treated by 1, 10 and 100 mmol/L trehalose by 0 mmol/L trehalose, 62 percent, 123 percent, 37 percent and 213 percent, 368 percent and 188 percent are respectively improved compared with those of the plant treated by 0.0 mmol/L trehalose, wherein the plant treated by 10 mmol/L trehalose has the best effect, expresses strong CdCl 2 stress resistance, and is characterized in that the plant material of the okra seedlings treated by trehalose has the advantages of increasing the trehalose content, increasing the proline content, increasing the chlorophyll a content, increasing the chlorophyll b content, increasing the total chlorophyll content, increasing the total carotenoid content, increasing the relative water content, increasing the SOD activity, increasing the CAT activity, increasing the GPX activity, increasing the GR activity, increasing the APX activity, increasing the POD activity, decreasing the electrolyte permeability and decreasing the H 2O2 content and the malondialdehyde content.
The result shows that the trehalose has important application in relieving the stress effect of CdCl 2 on okra seedlings, the trehalose can promote the growth of okra seedlings under the stress of CdCl 2 by using the trehalose crop relieving agent with a certain concentration, the tolerance of okra seedlings to the stress of CdCl 2 is improved, the relation of the trehalose and the CdCl 2 stress of okra in osmotic regulation, leaf damage, photosynthesis, an antioxidant enzyme system and a cell membrane system is researched, the action mechanism of the trehalose on plants for resisting heavy metal stress is discussed, and a theoretical basis is provided for the safe production of the trehalose in okra.
Drawings
FIG. 1 effect of different concentrations (0, 1, 10 and 100 mmol/L) of trehalose on okra seed germination rate and seedling fresh weight under normal conditions and 0.1 mmol/LCdCl 2 stress of the invention.
FIG. 2 identification of the remission effect resistance of trehalose at different concentrations (0, 1, 10 and 100 mmol/L) under normal conditions and 0.1 mmol/LCdCl 2 stress according to the invention on okra seedlings.
FIG. 3 effect of 10 mmol/L trehalose on okra seedlings on trehalose and proline content under normal conditions and 0.1 mmol/L CdCl 2 stress of the invention.
FIG. 4 effect of 10 mmol/L trehalose on photosynthetic pigment content (chlorophyll a, total chlorophyll and total carotenoids) of okra seedlings under normal conditions and 0.1 mmol/LCdCl 2 stress of the present invention.
FIG. 5 effects of 10. 10 mmol/L trehalose on malondialdehyde content, electrolyte permeability, relative water content and H 2O2 content of okra seedlings under normal conditions and 0.1 mmol/LCdCl 2 stress of the present invention.
FIG. 6 effect of 10 mmol/L trehalose on the activity of key enzyme (SOD, GPX, CAT, GR, APX, POD) of okra seedling ROS scavenging system under normal conditions and 0.1 mmol/LCdCl 2 stress of the present invention.
Description of the embodiments
The following examples facilitate a better understanding of the present invention, but are not intended to limit the same. The experimental methods in the following examples are conventional methods unless otherwise specified. The test materials used in the examples described below, unless otherwise specified, were commercially available from conventional biochemical reagents. The quantitative tests in the following examples were all set up in triplicate and the results averaged.
In the examples, okra (Abelmoschus esculentus) variety Taiwan five-Fu was used as the test material. The okra variety taiwan wufu, reference 【Feibing Wang, Gaolei Ren, Fengsheng Li, Sitong Qi, Yan Xu, Bowen Wang, Yulin Yang, Yuxiu Ye, Qing Zhou, Xinhong Chen. A chalcone synthase gene AeCHS from Abelmoschus esculentus regulates flavonoid accumulation and abiotic stress tolerance in transgenic Arabidopsis. 2018, 40:97】, was saved by the Huaiyin institute of engineering life sciences and food engineering, academy of plant production and processing practice education center laboratory.
Trehalose (Trehalose, abbreviated as Tre) purchased from Sigma company.
The Cd donor was cadmium chloride (CdCl 2·2H2 O), purchased from Sigma.
Example 1 exogenous trehalose Regulation of CdCl 2 stress resistance assay of okra seedlings
1. Analysis of CdCl 2 stress-resistant germination rate of okra seeds treated by trehalose with different concentrations
Referring to Wang Yonghui et al (2014) [ Wang Yonghui, chen Jianping, zhang Peitong, cai Liwang, shi Qinghua, wang Haiyang. Abelmoschus esculentus salt tolerance material screening and analysis of salt tolerance during germination, southwest agricultural report, 2014,27 (2): 788-792 ], abelmoschus esculentus seed material was selected from 50 full seeds, sterilized 5min with 5% hypochlorous acid solution, washed 3 times with distilled water, soaked in clear water and 0.1mmol/L CdCl 2 solution at room temperature for 24 hours, respectively, and then placed in a transparent plastic fresh-keeping box with two layers of filter paper having an inner diameter of 12 cm, and then subjected to treatments of different concentrations (0, 1, 10 and 100 mmol/L) of trehalose, respectively, including (i) adding 0, 1, 10 and 100 mmol/L of trehalose with distilled water under normal conditions, (ii) adding 0, 1, 10 and 100 mmol/L of trehalose under conditions of 0.1mmol/L CdCl 2, respectively. Each treatment was repeated six times. The fresh-keeping box is placed in a climatic chamber for germination, the white/night temperature is 28 ℃ per 25 ℃, the daytime/black day is 12 h, and the humidity is 60% +/-1%. And taking the radicle length of 0.2 mm as a seed germination mark, and counting the germination number of the seeds on the 5 th day. Seed germination = (number of germinated seeds/total number of test seeds on day 5) ×100%.
The results show that the germination rate test results of the stress on CdCl 2 of 0.1 mmol/L show that the germination rate (figure 1A) and fresh weight (figure 1B) of okra seeds treated by the trehalose of figures 1,1 and 10 mmol/L are significantly higher than those of okra seeds treated by the trehalose of 0 and 10 mmol/L, and the germination rate is improved by 51%, 130% and 47% and 118% respectively compared with that of okra seedlings treated by the trehalose of 0 mmol/L, wherein the okra seedlings treated by the trehalose of 10 mmol/L have the best effect.
2. Analysis of CdCl 2 stress resistance of okra seedlings treated with trehalose at different concentrations
Referring to the method of Li et al （2022）【Hengpeng Li, Shasha Yang, Wenya Wu, Chunyan Wang, Yanyang Li, Chenzhong Wan, Yuxiu Ye, Xinhong Chen, Zunxin Wang, Laibao Hu, Feibing Wang. Physiological and biochemical mechanisms of improving salt and drought tolerance in okra plants based on applied attapulgite clay. Advances in Biochemistry, 2022, 10:1-10】, abelmoschus manihot seedlings of Taiwan were transplanted into plastic pots (19 cm in diameter) containing a mixture of turf, humus and vermiculite (1:1:1, v/v/v) in the greenhouse. All seedlings were thoroughly watered with half Hoagland solution for 4 weeks until the seedlings developed new leaves. Subsequently, cdCl 2 stress and trehalose treatments were performed, including (i) normally, irrigation with fresh water every pot of okra seedling plants, once every 2 days for 4 weeks, while spraying aqueous solutions containing 0, 1, 10 and 100 mmol/L of trehalose every day, 3 times per day. (ii) CdCl 2 stress treatment, irrigation with 100ml of 0.1 mmol/L CdCl 2 solution was performed on each pot of okra seedling plants once every 2 days for 4 weeks, while spraying aqueous solutions containing 0, 1, 10 and 100 mmol/L trehalose every day, 3 times a day.
The result shows that the result of the 0.1 mmol/L CdCl 2 stress potting experiment shows that, as shown in FIG. 2, the okra seedling plants treated by exogenous trehalose grow well and have improved resistance, the fresh weight and dry weight of the okra seedlings treated by the exogenous trehalose are obviously higher than those of the okra seedlings treated by the exogenous trehalose, which are respectively 62%, 123%, 37% and 213%, 368% and 188% higher than those of the okra seedlings treated by the exogenous trehalose, and the okra seedlings treated by the exogenous trehalose are respectively improved by 0. mmol/L compared with those of the okra seedlings treated by the exogenous trehalose, wherein the okra seedling plants treated by the exogenous trehalose have the best effect, and the okra seedling treated by the exogenous trehalose expresses very strong CdCl 2 stress resistance.
Therefore, the phenotype identification result shows that the okra plant treated by the exogenous 10 mmol/L trehalose has the optimal CdCl 2 stress resistance phenotype, and is used for analyzing a physiological and biochemical mechanism for inducing CdCl 2 stress resistance.
Example 2 determination of physiological and Biochemical indicators of resistance of okra seedlings to CdCl 2 stress
Trehalose is a safe and reliable natural sugar, is non-reducing sugar formed by two glucose molecules with 1, 1-glycosidic bond, is disaccharide with the most stable property in sugar, has low content in plant body, and can stabilize the structure and function of protein by regulating the content of trehalose in plant under adverse conditions. Research shows that trehalose is a penetration regulating substance, not only can protect an antioxidant enzyme system and subcellular structures, but also can mediate penetration regulation and activate ROS clearance, and can relieve damage to plants caused by stress.
Determination methods reference 【Tao Jiang, Hong Zhai, Feibing Wang, Huanan Zhou, Zengzhi Si, Shaozhen He, Qingchang Liu. Cloning and characterization of a salt tolerance-associated gene encoding trehalose-6-phosphate synthase in sweetpotato. Journal of Integrative Agriculture, 2014, 13(8):1651-1661】, detects trehalose content of okra seedling plants. Okra plants are non-stressed in the above potted plant identification, were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves, and were treated with 0.1. 0.1 mmol/L CdCl 2 in the above potted plant identification, and were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves. The experiment was repeated three times and the results averaged.
The experimental results are shown in FIG. 3A (Normal is a blank control, cdCl 2 stress is heavy metal stress). The result shows that after 0.1 mmol/L CdCl 2 stress treatment, the trehalose content in the okra plants treated by the exogenous trehalose is significantly higher than that of the okra plants not treated by the trehalose.
Under normal conditions, plants have very low free proline content, but when subjected to salt, drought and other stresses, free amino acids accumulate in large quantities, and the accumulation index is related to stress resistance of the plants. Thus, proline can be used as a biochemical indicator of stress resistance in plants.
Assay reference 【Feibing Wang, Weili Kong, Gary Wong, Lifeng Fu, Rihe Peng, Zhenjun Li, Quanhong Yao. AtMYB12 regulates flavonoids accumulation and abiotic stress tolerance in transgenic Arabidopsis thaliana. Molecular Genetics and Genomics, 2016, 291:1545-1559】, detects proline content in okra seedling plants. Okra plants are non-stressed in the above potted plant identification, were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves, and were treated with 0.1. 0.1 mmol/L CdCl 2 in the above potted plant identification, and were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves. The experiment was repeated three times and the results averaged.
The experimental results are shown in FIG. 3B (Normal is a blank control, cdCl 2 stress is heavy metal stress). The result shows that the proline content in the okra plants treated by the exogenous trehalose is obviously higher than that of the okra plants not treated by the trehalose after the stress treatment of 0.1 mmol/L CdCl 2.
Chloroplasts are important organelles of plants for carrying out physiological processes such as photosynthesis and adverse stress reaction, and photosynthetic pigments in the chloroplasts comprise chlorophyll and carotenoid, wherein the chlorophyll is divided into chlorophyll a and chlorophyll b. Adverse stress affects photosynthesis of plants, and increases the number of free radicals in chloroplasts, thereby destroying chlorophyll, affecting photosynthesis of plants, and its photosynthetic pigment content reflects the ability of plants to perform photosynthesis.
Assay reference 【Lichtenthaler Hartmut K, Buschmann Claus. Chlorophylls and carotenoids: measurement and characterization by UV-VIS spectroscop. Current Protocols in Food Analytical Chemistry, 2001, 1】, detects photosynthetic pigment content of okra seedling plants. Okra plants are non-stressed in the above potted plant identification, were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves, and were treated with 0.1. 0.1 mmol/L CdCl 2 in the above potted plant identification, and were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves. The experiment was repeated three times and the results averaged.
The experimental results are shown in FIG. 4 (Normal is a blank control, cdCl 2 stress is heavy metal stress). The results showed that chlorophyll a (fig. 4A), chlorophyll B (fig. 4B), total chlorophyll (fig. 4C) and total carotenoid content (fig. 4D) were significantly higher in the exogenous trehalose-treated okra plants after 0.1 mmol/L CdCl 2 stress treatment than in okra plants not treated with trehalose.
Plant organ aging or injury in adverse circumstances often occurs, and Malondialdehyde (MDA) is the final decomposition product of membrane lipid peroxidation, and its content can reflect the degree of adverse circumstances injury of plants, i.e. the higher the MDA content, the greater the degree of adverse circumstances injury of plants.
Assay reference 【Feibing Wang, Weili Kong, Gary Wong, Lifeng Fu, Rihe Peng, Zhenjun Li, Quanhong Yao. AtMYB12 regulates flavonoids accumulation and abiotic stress tolerance in transgenic Arabidopsis thaliana. Molecular Genetics and Genomics, 2016, 291:1545-1559】, detects MDA content of okra seedling plants. Okra plants are non-stressed in the above potted plant identification, were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves, and were treated with 0.1. 0.1 mmol/L CdCl 2 in the above potted plant identification, and were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves. The experiment was repeated three times and the results averaged.
The experimental results are shown in FIG. 5A (Normal is a blank control, cdCl 2 stress is heavy metal stress). The result shows that after 0.1 mmol/L CdCl 2 stress treatment, the MDA content in the okra plants treated by the exogenous trehalose is significantly lower than that of the okra plants not treated by the trehalose.
The permeability of the electrolyte is an index for measuring the permeability of the plant cell membrane, and the lower the value is, the more complete the cell membrane is, and the function is good.
Determination methods reference 【Yujia Liu, Xiaoyu Ji, Xianguang Nie, Min Qu, Lei Zheng, Zilong Tan, Huimin Zhao, Lin Huo,Shengnan Liu, Bing Zhang, Yucheng Wang. Arabidopsis AtbHLH112 regulates the expression of genes involved in abiotic stress tolerance by binding to their E-box and GCG-box motifs. New Phytologist, 2015, 207:692-709】, detects electrolyte permeability of okra seedling plants. Okra plants are non-stressed in the above potted plant identification, were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves, and were treated with 0.1. 0.1 mmol/L CdCl 2 in the above potted plant identification, and were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves. The experiment was repeated three times and the results averaged.
The experimental results are shown in FIG. 5B (Normal is a blank control, cdCl 2 stress is heavy metal stress). The result shows that the electrolyte permeability of the okra plants treated by the exogenous trehalose is obviously lower than that of the okra plants not treated by the trehalose after the stress treatment of 0.1 mmol/L CdCl 2.
The relative water content is an important index reflecting the water content condition of plants, the relative water content of plant tissues is positively correlated with the stress resistance of plants, and the relative water content of cells reflects the stress degree of cells.
Assay reference 【Yufeng Yang, Shikai Guan, Hong Zhai, Shaozhen He, Qingchang Liu. Development and evaluation of a storage root-bearing sweetpotato somatic hybrid between Ipomoea batatas (L.) Lam. and I. trilobaL. Plant Cell, Tissue and Organ Culture, 2009, 99:83-89】, detects the relative water content of okra seedling plants. Okra plants are non-stressed in the above potted plant identification, were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves, and were treated with 0.1. 0.1 mmol/L CdCl 2 in the above potted plant identification, and were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves. The experiment was repeated three times and the results averaged.
The experimental results are shown in FIG. 5C (Normal is a blank control, cdCl 2 stress is heavy metal stress). The result shows that after 0.1 mmol/L CdCl 2 stress treatment, the relative water content in the okra plants treated by the exogenous trehalose is significantly higher than that of the okra plants not treated by the trehalose.
7. Determination of H 2O2 content
H 2O2 accumulates in plants under stress or in senescence due to enhanced in vivo active oxygen metabolism. H 2O2 can oxidize biomacromolecules such as nucleic acids, proteins and the like in cells directly or indirectly, and damage cell membranes, thereby accelerating aging and disintegration of cells. Thus, the higher the content of H 2O2, the greater the extent to which the plant suffers from stress injury.
Assay reference 【Feibing Wang, Weili Kong, Gary Wong, Lifeng Fu, Rihe Peng, Zhenjun Li, Quanhong Yao. AtMYB12 regulates flavonoids accumulation and abiotic stress tolerance in transgenic Arabidopsis thaliana. Molecular Genetics and Genomics, 2016, 291:1545-1559】, detects H 2O2 content of okra seedling plants. Okra plants are non-stressed in the above potted plant identification, were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves, and were treated with 0.1. 0.1 mmol/L CdCl 2 in the above potted plant identification, and were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves. The experiment was repeated three times and the results averaged.
The experimental results are shown in FIG. 5D (Normal is a blank control, cdCl 2 stress is heavy metal stress). The result shows that after 0.1 mmol/L CdCl 2 stress treatment, the H 2O2 content in the okra plants treated by the exogenous trehalose is significantly lower than that in the okra plants not treated by the trehalose.
Superoxide dismutase (SOD) activity can be used as a physiological and biochemical index of plant stress resistance. The lower the activity of SOD, the greater the extent to which the plant suffers from stress injury.
Assay reference 【Feibing Wang, Weili Kong, Gary Wong, Lifeng Fu, Rihe Peng, Zhenjun Li, Quanhong Yao. AtMYB12 regulates flavonoids accumulation and abiotic stress tolerance in transgenic Arabidopsis thaliana. Molecular Genetics and Genomics, 2016, 291:1545-1559】, detects SOD activity in okra seedling plants. Okra plants are non-stressed in the above potted plant identification, were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves, and were treated with 0.1. 0.1 mmol/L CdCl 2 in the above potted plant identification, and were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves. The experiment was repeated three times and the results averaged.
The experimental results are shown in FIG. 6A (Normal is a blank control, cdCl 2 stress is heavy metal stress). The result shows that the SOD activity in the okra plants treated by the exogenous trehalose is obviously higher than that of the okra plants not treated by the trehalose after the stress treatment of 0.1 mmol/L CdCl 2.
Glutathione Peroxidase (GPX) is an important peroxidase widely existing in the body, is a key enzyme for removing active oxygen in the body, and plays an important role in plant stress resistance reaction.
Assay reference 【Hong Zhai, Feibing Wang, Zengzhi Si, Jinxi Huo, Lei Xing, Yanyan An, Shaozhen He, Qingchang Liu. A myo-inositol-1-phosphate synthase gene, IbMIPS1, enhances salt and drought tolerance and stem nematode resistance in transgenic sweetpotato. Plant Biotechnology Journal, 2016, 14:592-602】, detects GPX activity in okra seedling plants. Okra plants are non-stressed in the above potted plant identification, were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves, and were treated with 0.1. 0.1 mmol/L CdCl 2 in the above potted plant identification, and were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves. The experiment was repeated three times and the results averaged.
The experimental results are shown in FIG. 6B (Normal is a blank control, cdCl 2 stress is heavy metal stress). The result shows that GPX activity in okra plants treated by exogenous trehalose is significantly higher than that of okra plants not treated by trehalose after stress treatment of 0.1 mmol/L CdCl 2.
Catalase (CAT) is commonly found in plant tissues and is one of important protective enzymes, and the function of the catalase is to remove H 2O2 generated in metabolism so as to avoid oxidative damage of H 2O2 accumulation on cells, so that the activity of the catalase is related to stress resistance of plants.
Assay reference 【Yufeng Yang, Shikai Guan, Hong Zhai, Shaozhen He, Qingchang Liu. Development and evaluation of a storage root-bearing sweetpotato somatic hybrid between Ipomoea batatas (L.) Lam. and I. trilobaL. Plant Cell, Tissue and Organ Culture, 2009, 99:83-89】, detects CAT activity in okra seedling plants. Okra plants are non-stressed in the above potted plant identification, were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves, and were treated with 0.1. 0.1 mmol/L CdCl 2 in the above potted plant identification, and were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves. The experiment was repeated three times and the results averaged.
The experimental results are shown in FIG. 6C (Normal is a blank control, cdCl 2 stress is heavy metal stress). The result shows that after 0.1 mmol/L CdCl 2 stress treatment, CAT activity in okra plants treated by exogenous trehalose is significantly higher than that of okra plants not treated by trehalose.
Glutathione Reductase (GR) is a ubiquitous flavin enzyme involved in the defense system against cellular stress, and its activity is related to stress resistance of plants.
Assay reference 【Wenjin Zhang, Zhicai Xie, Lianhong Wang, Ming Li, Duoyong Lang, Xinhui Zhang. Silicon alleviates salt and drought stress of Glycyrrhiza uralensis seedling by altering antioxidant metabolism and osmotic adjustment. Journal of Plant Research, 2017, 130:611-624】, detects GR activity in okra seedling plants. Okra plants are non-stressed in the above potted plant identification, were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves, and were treated with 0.1. 0.1 mmol/L CdCl 2 in the above potted plant identification, and were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves. The experiment was repeated three times and the results averaged.
The experimental results are shown in FIG. 6D (Normal is a blank control, cdCl 2 stress is heavy metal stress). The result shows that after 0.1 mmol/L CdCl 2 stress treatment, the GR activity in the okra plants treated by the exogenous trehalose is significantly higher than that of the okra plants not treated by the trehalose.
Ascorbate Peroxidase (APX) is one of the important antioxidant enzymes in plant active oxygen metabolism, especially the key enzyme for removing H 2O2 in chloroplasts, and is also the main enzyme class of vitamin C metabolism. APX activity can be used as a physiological and biochemical index of plant stress resistance, and the level of the activity is related to plant stress resistance.
Assay reference 【Hong Zhai, Feibing Wang, Zengzhi Si, Jinxi Huo, Lei Xing, Yanyan An, Shaozhen He, Qingchang Liu. A myo-inositol-1-phosphate synthase gene, IbMIPS1, enhances salt and drought tolerance and stem nematode resistance in transgenic sweetpotato. Plant Biotechnology Journal, 2016, 14:592-602】, detects APX activity in okra seedling plants. Okra plants are non-stressed in the above potted plant identification, were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves, and were treated with 0.1. 0.1 mmol/L CdCl 2 in the above potted plant identification, and were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves. The experiment was repeated three times and the results averaged.
The experimental results are shown in FIG. 6E (Normal is a blank control, cdCl 2 stress is heavy metal stress). The result shows that the APX activity in the okra plants treated by the exogenous trehalose is obviously higher than that of the okra plants not treated by the trehalose after the stress treatment of 0.1 mmol/L CdCl 2.
Peroxidase (POD) activity can be used as a physiological and biochemical indicator of plant stress resistance. The lower the activity of POD, the greater the extent to which the plant suffers from stress injury.
Assay reference 【Feibing Wang, Weili Kong, Gary Wong, Lifeng Fu, Rihe Peng, Zhenjun Li, Quanhong Yao. AtMYB12 regulates flavonoids accumulation and abiotic stress tolerance in transgenic Arabidopsis thaliana. Molecular Genetics and Genomics, 2016, 291:1545-1559】, detects POD activity in okra seedling plants. Okra plants are non-stressed in the above potted plant identification, were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves, and were treated with 0.1. 0.1 mmol/L CdCl 2 in the above potted plant identification, and were sprayed with 10 mmol/L aqueous trehalose solution for 2 weeks of okra seedling leaves. The experiment was repeated three times and the results averaged.
The experimental results are shown in FIG. 6, F (Normal is a blank control, cdCl 2 stress is heavy metal stress). The result shows that POD activity in okra plants treated by exogenous trehalose is significantly higher than that of okra plants not treated by trehalose after stress treatment of 0.1 mmol/L CdCl 2.
The measurement result of the physiological and biochemical index shows that the exogenous trehalose treatment obviously improves the CdCl 2 stress resistance of the okra plant. The relation of the trehalose and the CdCl 2 under the stress of the okra is studied, the action mechanism of the trehalose in the heavy metal stress resistance of plants is discussed, and a theoretical basis is provided for the safe production of the trehalose in the okra.
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