CN110338024A - Composition of Brassinolide and Salicylic Acid to Alleviate Oxidative Stress of Tobacco by Cadmium - Google Patents
Composition of Brassinolide and Salicylic Acid to Alleviate Oxidative Stress of Tobacco by Cadmium Download PDFInfo
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- CN110338024A CN110338024A CN201910614195.1A CN201910614195A CN110338024A CN 110338024 A CN110338024 A CN 110338024A CN 201910614195 A CN201910614195 A CN 201910614195A CN 110338024 A CN110338024 A CN 110338024A
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- YGSDEFSMJLZEOE-UHFFFAOYSA-N salicylic acid Chemical compound OC(=O)C1=CC=CC=C1O YGSDEFSMJLZEOE-UHFFFAOYSA-N 0.000 title claims abstract description 76
- 235000002637 Nicotiana tabacum Nutrition 0.000 title claims abstract description 43
- IXVMHGVQKLDRKH-VRESXRICSA-N Brassinolide Natural products O=C1OC[C@@H]2[C@@H]3[C@@](C)([C@H]([C@@H]([C@@H](O)[C@H](O)[C@H](C(C)C)C)C)CC3)CC[C@@H]2[C@]2(C)[C@@H]1C[C@H](O)[C@H](O)C2 IXVMHGVQKLDRKH-VRESXRICSA-N 0.000 title claims abstract description 38
- FJKROLUGYXJWQN-UHFFFAOYSA-N papa-hydroxy-benzoic acid Natural products OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 title claims abstract description 36
- 229960004889 salicylic acid Drugs 0.000 title claims abstract description 36
- 239000000203 mixture Substances 0.000 title claims abstract description 24
- 229910052793 cadmium Inorganic materials 0.000 title claims abstract description 14
- BDOSMKKIYDKNTQ-UHFFFAOYSA-N cadmium atom Chemical compound [Cd] BDOSMKKIYDKNTQ-UHFFFAOYSA-N 0.000 title claims abstract description 14
- 230000036542 oxidative stress Effects 0.000 title claims abstract description 11
- IXVMHGVQKLDRKH-KNBKMWSGSA-N brassinolide group Chemical group [C@@H](C)([C@@H](O)[C@H](O)[C@@H](C)C(C)C)[C@H]1CC[C@H]2[C@@H]3COC(=O)[C@H]4C[C@H](O)[C@H](O)C[C@]4(C)[C@H]3CC[C@]12C IXVMHGVQKLDRKH-KNBKMWSGSA-N 0.000 title description 27
- 244000061176 Nicotiana tabacum Species 0.000 title description 2
- 241000208125 Nicotiana Species 0.000 claims abstract description 41
- IXVMHGVQKLDRKH-YEJCTVDLSA-N (22s,23s)-epibrassinolide Chemical compound C1OC(=O)[C@H]2C[C@H](O)[C@H](O)C[C@]2(C)[C@H]2CC[C@]3(C)[C@@H]([C@H](C)[C@H](O)[C@@H](O)[C@H](C)C(C)C)CC[C@H]3[C@@H]21 IXVMHGVQKLDRKH-YEJCTVDLSA-N 0.000 claims description 5
- 150000002596 lactones Chemical class 0.000 claims 1
- 241000196324 Embryophyta Species 0.000 abstract description 22
- 125000003861 brassinosteroid group Chemical group 0.000 abstract description 6
- 230000006378 damage Effects 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 4
- 238000002474 experimental method Methods 0.000 abstract description 3
- 150000001647 brassinosteroids Chemical class 0.000 abstract description 2
- 239000007921 spray Substances 0.000 description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 10
- 239000002689 soil Substances 0.000 description 9
- YKYOUMDCQGMQQO-UHFFFAOYSA-L cadmium dichloride Chemical compound Cl[Cd]Cl YKYOUMDCQGMQQO-UHFFFAOYSA-L 0.000 description 8
- 238000005507 spraying Methods 0.000 description 8
- 230000012010 growth Effects 0.000 description 7
- 229920001213 Polysorbate 20 Polymers 0.000 description 6
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 6
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 6
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 description 5
- 230000000052 comparative effect Effects 0.000 description 5
- 229940118019 malondialdehyde Drugs 0.000 description 5
- 238000012545 processing Methods 0.000 description 5
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 4
- 238000011161 development Methods 0.000 description 4
- 229910001385 heavy metal Inorganic materials 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 229910052760 oxygen Inorganic materials 0.000 description 4
- 239000001301 oxygen Substances 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 230000008635 plant growth Effects 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 230000003859 lipid peroxidation Effects 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 239000010451 perlite Substances 0.000 description 2
- 235000019362 perlite Nutrition 0.000 description 2
- 230000035882 stress Effects 0.000 description 2
- 239000010455 vermiculite Substances 0.000 description 2
- 235000019354 vermiculite Nutrition 0.000 description 2
- 229910052902 vermiculite Inorganic materials 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- CBENFWSGALASAD-UHFFFAOYSA-N Ozone Chemical compound [O-][O+]=O CBENFWSGALASAD-UHFFFAOYSA-N 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 230000036579 abiotic stress Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000032823 cell division Effects 0.000 description 1
- 239000013043 chemical agent Substances 0.000 description 1
- 229930002875 chlorophyll Natural products 0.000 description 1
- 235000019804 chlorophyll Nutrition 0.000 description 1
- ATNHDLDRLWWWCB-AENOIHSZSA-M chlorophyll a Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 ATNHDLDRLWWWCB-AENOIHSZSA-M 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000006735 deficit Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- 150000002148 esters Chemical group 0.000 description 1
- 239000003630 growth substance Substances 0.000 description 1
- 239000004009 herbicide Substances 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 239000012452 mother liquor Substances 0.000 description 1
- 238000005502 peroxidation Methods 0.000 description 1
- 150000002989 phenols Chemical class 0.000 description 1
- 230000000243 photosynthetic effect Effects 0.000 description 1
- 230000008121 plant development Effects 0.000 description 1
- 230000008638 plant developmental process Effects 0.000 description 1
- 230000008636 plant growth process Effects 0.000 description 1
- 239000003375 plant hormone Substances 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 231100000719 pollutant Toxicity 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000007226 seed germination Effects 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/45—Tobacco
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
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- Life Sciences & Earth Sciences (AREA)
- Biodiversity & Conservation Biology (AREA)
- Ecology (AREA)
- Forests & Forestry (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Cultivation Of Plants (AREA)
Abstract
本发明公开了油菜素内酯和水杨酸的组合物缓解镉对烟草氧化胁迫的应用，所述油菜素内酯和水杨酸的组合物按质量比为1:(0.7‑7.2)的比例由油菜素内酯和水杨酸组成。本发明的组合物无污染，对植物没有伤害。发明的组合物成本较低，实验证明油菜素内酯和水杨酸的组合物缓解镉对烟草氧化胁迫作用。The invention discloses the application of a composition of brassinosteroid and salicylic acid to relieve cadmium on tobacco oxidative stress, and the composition of brassinosteroid and salicylic acid has a mass ratio of 1:(0.7-7.2) Composed of brassinosteroids and salicylic acid. The composition of the invention is non-polluting and has no harm to plants. The cost of the invented composition is low, and experiments prove that the composition of brassinosteroid and salicylic acid alleviates the oxidative stress effect of cadmium on tobacco.
Description
技术领域technical field
本发明涉及植物生物学领域，具体涉及一种油菜素内酯和水杨酸的组合物缓解镉对烟草氧化胁迫的应用。The invention relates to the field of plant biology, in particular to the application of a composition of brassinosteroid and salicylic acid to alleviate the oxidative stress of cadmium on tobacco.
背景技术Background technique
镉(Cd)是生物毒性最强的重金属，被称为“五毒之首”，其毒害在当前是一个愈来愈严峻的农业和环境问题。随着工业化和城市化的发展，农田土壤和水体重金属镉累积超标问题日益严重，含镉的污染物通过各种途径进入土壤，Cd不是植物生长所必需的元素，由于其亲水性，容易在植物体内积累。大量研究表明，重金属Cd会严重损害植物代谢，如降低光合速率、抑制叶绿素合成、脂质过氧化、增强或抑制抗氧化酶活性等。Cadmium (Cd) is the most toxic heavy metal, known as "the first of the five poisons", and its toxicity is an increasingly serious agricultural and environmental problem at present. With the development of industrialization and urbanization, the accumulation of heavy metal cadmium in farmland soil and water has become increasingly serious. Pollutants containing cadmium enter the soil through various channels. Cd is not an essential element for plant growth. accumulation in plants. A large number of studies have shown that heavy metal Cd can seriously damage plant metabolism, such as reducing photosynthetic rate, inhibiting chlorophyll synthesis, lipid peroxidation, enhancing or inhibiting the activity of antioxidant enzymes, etc.
油菜素内酯(BR)在植物体内广泛分布，主要存在于花和未成熟的种子中，在植物生长发育过程中，其含量的微小变化都会对植物生长发育产生重要影响，第十六届国际植物生长物质会议上BR被认定为第六大植物激素。研究表明，BR能够促进植物细胞伸长、细胞分裂、能充分激发植物内在潜能，促进作物生长和增加作物产量，提高植物的耐冷性，提高植物的抗病、抗盐能力，使植物的耐逆性增强，可减轻除草剂等化学药剂对植物的伤害。Brassinolide (BR) is widely distributed in plants and mainly exists in flowers and immature seeds. During the process of plant growth and development, small changes in its content will have an important impact on plant growth and development. The 16th International BR was identified as the sixth largest plant hormone in the Plant Growth Substance Conference. Studies have shown that BR can promote plant cell elongation, cell division, fully stimulate the inherent potential of plants, promote crop growth and increase crop yield, improve plant cold tolerance, improve plant resistance to disease and salt, and make plants resistant to stress Sex enhancement, which can reduce the damage of chemical agents such as herbicides to plants.
水杨酸(SA)是植物体内普遍存在的一种酚类化合物，是细胞内的信号传递分子，对植物的生长发育具有多种生理调节效应，不仅能增强植物抵抗真菌、细菌和病毒等生物胁迫的能力，而且还影响着植物对重金属、臭氧、紫外辐射、低温、热激、水分亏缺和盐害等非生物胁迫的抗性。Salicylic acid (SA) is a phenolic compound ubiquitous in plants. It is a signal transduction molecule in cells. It has a variety of physiological regulatory effects on the growth and development of plants. It can not only enhance the resistance of plants to organisms such as fungi, bacteria and viruses It also affects the resistance of plants to abiotic stresses such as heavy metals, ozone, ultraviolet radiation, low temperature, heat shock, water deficit and salt damage.
目前，现有报道大多是关于BR与SA分别与植物的抗旱性、抗盐性、抗寒性等的研究，尚未有BR与SA共同作用缓解镉对烟草氧化胁迫的应用的报道。At present, most of the existing reports are about the research on BR and SA respectively and the drought resistance, salt resistance, cold resistance of plants, etc. There is no report on the application of BR and SA to alleviate the oxidative stress of cadmium on tobacco.
发明内容Contents of the invention
本发明的目的在于克服现有技术的不足，提供油菜素内酯和水杨酸的组合物缓解镉对烟草氧化胁迫的应用。The purpose of the present invention is to overcome the deficiencies of the prior art, and provide a composition of brassinosteroid and salicylic acid to alleviate the oxidative stress of cadmium on tobacco.
本发明的技术方案概述如下：Technical scheme of the present invention is summarized as follows:
油菜素内酯和水杨酸的组合物缓解镉对烟草氧化胁迫的应用，所述油菜素内酯和水杨酸的组合物按质量比为1:(0.7-7.2)的比例由油菜素内酯和水杨酸组成。The composition of brassinolide and salicylic acid alleviates the application of cadmium to tobacco oxidative stress, and the composition of said brassinolide and salicylic acid is composed of brassinosteroids in a mass ratio of 1: (0.7-7.2). Composition of esters and salicylic acid.
优选地，油菜素内酯和水杨酸的质量比为1:2.2。Preferably, the mass ratio of brassinolide and salicylic acid is 1:2.2.
本发明的优点:Advantage of the present invention:
(1)本发明的组合物无污染，对植物没有伤害。(1) The composition of the present invention is non-polluting and has no harm to plants.
(2)本发明的成本较低，实验证明油菜素内酯和水杨酸的组合物缓解镉对烟草氧化胁迫作用。(2) The cost of the present invention is relatively low, and the experiment proves that the composition of brassinosteroid and salicylic acid relieves the oxidative stress effect of cadmium on tobacco.
附图说明Description of drawings
图1是烟草在不同情况下H2O2含量。Figure 1 is the content of H 2 O 2 in tobacco under different conditions.
图2是烟草在不同情况下O2 -含量。Figure 2 is the O 2 -content of tobacco under different conditions.
图3是烟草在不同情况下丙二醛含量。Figure 3 shows the MDA content of tobacco under different conditions.
图中，WT为对照例1野生型烟草；WT+Cd为对照例2；WT+Cd+BR为对照例3；WT+Cd+SA为对照例4；WT+Cd+BR+SA为实施例4。In the figure, WT is the wild-type tobacco of the control example 1; WT+Cd is the control example 2; WT+Cd+BR is the control example 3; WT+Cd+SA is the control example 4; WT+Cd+BR+SA is the embodiment 4.
具体实施方式Detailed ways
本发明以野生型烟草为例，是为了使本领域的技术人员能够更好地理解本发明，但并不对本发明作任何限制。The present invention takes wild-type tobacco as an example to enable those skilled in the art to better understand the present invention, but does not limit the present invention in any way.
野生型烟草种子、营养土、蛭石、珍珠岩均为市售。Wild-type tobacco seeds, nutrient soil, vermiculite, and perlite are all commercially available.
CdCl2试剂购自天津市元立化工有限公司，纯度为分析纯。CdCl 2 reagent was purchased from Tianjin Yuanli Chemical Co., Ltd., and its purity was analytically pure.
BR试剂(油菜素内酯)购自天津鼎国生物公司，纯度为分析纯。BR reagent (brassinolide) was purchased from Tianjin Dingguo Biological Company, and its purity was analytically pure.
SA试剂(水杨酸)购自天津鼎国生物公司，纯度98％。SA reagent (salicylic acid) was purchased from Tianjin Dingguo Biological Company with a purity of 98%.
吐温20购自天津鼎国生物公司。Tween 20 was purchased from Tianjin Dingguo Biological Company.
生理指标的测定:H2O2含量、O2 -含量和丙二醛含量通过检测试剂盒测定。Determination of physiological indicators: H 2 O 2 content, O 2 - content and malondialdehyde content were determined by detection kits.
下面结合具体实施例对本发明作进一步的说明。The present invention will be further described below in conjunction with specific examples.
实施例1Example 1
油菜素内酯和水杨酸的组合物，由油菜素内酯和水杨酸组成。油菜素内酯和水杨酸的质量比为1:2.2。A composition of brassinolide and salicylic acid, consisting of brassinolide and salicylic acid. The mass ratio of brassinolide and salicylic acid is 1:2.2.
实施例2Example 2
油菜素内酯和水杨酸的组合物，由油菜素内酯和水杨酸组成。油菜素内酯和水杨酸的质量比为1:0.7。A composition of brassinolide and salicylic acid, consisting of brassinolide and salicylic acid. The mass ratio of brassinolide and salicylic acid is 1:0.7.
实施例3Example 3
油菜素内酯和水杨酸的组合物，由油菜素内酯和水杨酸组成。油菜素内酯和水杨酸的质量比为1:7.2。A composition of brassinolide and salicylic acid, consisting of brassinolide and salicylic acid. The mass ratio of brassinolide and salicylic acid is 1:7.2.
实施例4Example 4
油菜素内酯和水杨酸的组合物缓解镉对烟草氧化胁迫的应用，包括以下步骤：The composition of brassinolide and salicylic acid alleviates the application of cadmium to tobacco oxidative stress, comprising the following steps:
(1)进行种子萌发：(1) Carry out seed germination:
挑取饱满均一成熟的野生型烟草种子，将种子播种于装有营养土、蛭石和珍珠岩的塑料盆中，三者的体积比为2:2:1，在25℃/22℃昼夜、光暗16h/8h交替的温室中培养，待野生型烟草种子萌发、生长；Pick plump, uniform and mature wild-type tobacco seeds, sow the seeds in plastic pots filled with nutrient soil, vermiculite and perlite, the volume ratio of the three is 2:2:1, at 25°C/22°C day and night, light Cultivate in a greenhouse alternately dark for 16h/8h, and wait for wild-type tobacco seeds to germinate and grow;
(2)烟草幼苗的处理：(2) The processing of tobacco seedlings:
当烟草幼苗生长至5叶1心时，选取形态良好与长势一致的烟草幼苗，移入氯化镉浓度为20mg/kg的土壤中，每4d浇灌1次清水，每2d喷施1次实施例1的组合物的溶液(以乙醇为溶剂，配制油菜素内酯终浓度为1mg/mL、水杨酸终浓度为2.2mg/mL的母液，使用时，用水稀释至油菜素内酯终浓度为0.4mg/L、水杨酸终浓度为0.88mg/L)，喷洒时加入体积分数为0.2％的吐温20作为展开剂，用喷雾器在植株叶片正反两面均匀喷施，以叶片正反两面全部湿润且无液体滴下，以保证喷施充分、均匀。When the tobacco seedling grows to 5 leaves and 1 heart, select the tobacco seedling with good shape and consistent growth, move it into the soil with a cadmium chloride concentration of 20mg/kg, pour clear water once every 4d, and spray once every 2d Example 1 The solution of the composition (using ethanol as a solvent, prepare the mother liquor whose final concentration of brassinosteroid is 1mg/mL, and the final concentration of salicylic acid is 2.2mg/mL, when used, dilute with water to the final concentration of brassinosteroid is 0.4 mg/L, the final concentration of salicylic acid is 0.88mg/L), when spraying, add Tween 20 with a volume fraction of 0.2% as a developing agent, spray evenly on the front and back sides of the plant leaves with a sprayer, and use the front and back sides of the leaves to completely Wet and drip-free to ensure a full and even application.
(3)生理指标测定：(3) Determination of physiological indicators:
在烟草苗处理21d后，自上而下取第2片叶片，测定H2O2含量、O2 -含量和丙二醛含量。对照例1：After 21 days of tobacco seedling treatment, the second leaf was taken from top to bottom, and the H 2 O 2 content, O 2 - content and malondialdehyde content were measured. Comparative example 1:
步骤(1)同实施例4步骤(1)；Step (1) is the same as embodiment 4 step (1);
(2)烟草幼苗的处理：(2) The processing of tobacco seedlings:
当烟草幼苗生长至5叶1心时，选取形态良好与长势一致的烟草幼苗，移入土壤(不含镉)中，每4d浇灌1次清水，每2d喷施1次蒸馏水，喷洒时加入体积分数为0.2％的吐温20作为展开剂，用喷雾器在植株叶片正反两面均匀喷施，以叶片正反两面全部湿润且无液体滴下，以保证喷施充分、均匀。When the tobacco seedlings grow to 5 leaves and 1 heart, select tobacco seedlings with good shape and consistent growth, move them into the soil (without cadmium), water once every 4 days, spray distilled water once every 2 days, and add volume fraction when spraying Use 0.2% Tween 20 as a spreading agent, and spray evenly on both sides of the leaves of the plant with a sprayer, so that the front and back sides of the leaves are all wet and there is no liquid dripping to ensure sufficient and even spraying.
步骤(3)同实施例4步骤(3)。Step (3) is the same as embodiment 4 step (3).
对照例2：Comparative example 2:
步骤(1)同实施例4步骤(1)；Step (1) is the same as embodiment 4 step (1);
(2)烟草幼苗的处理：(2) The processing of tobacco seedlings:
当烟草幼苗生长至5叶1心时，选取形态良好与长势一致的烟草幼苗，移入氯化镉浓度为20mg/kg的土壤中，每4d浇灌1次清水，每2d喷施1次蒸馏水，喷洒时加入体积分数为0.2％的吐温20作为展开剂，用喷雾器在植株叶片正反两面均匀喷施，以叶片正反两面全部湿润且无液体滴下，以保证喷施充分、均匀。When the tobacco seedlings grow to 5 leaves and 1 heart, select tobacco seedlings with good shape and consistent growth, move them into the soil with a concentration of cadmium chloride of 20mg/kg, water once every 4d, spray distilled water once every 2d, spray When adding Tween 20 with a volume fraction of 0.2% as a developing agent, use a sprayer to spray evenly on the front and back of the plant leaves, so that the front and back of the leaves are all wet and there is no liquid dripping to ensure sufficient and even spraying.
步骤(3)同实施例4步骤(3)。Step (3) is the same as embodiment 4 step (3).
对照例3：Comparative example 3:
步骤(1)同实施例4步骤(1)；Step (1) is the same as embodiment 4 step (1);
(2)烟草幼苗的处理：(2) The processing of tobacco seedlings:
当烟草幼苗生长至5叶1心时，选取形态良好与长势一致的烟草幼苗，移入氯化镉浓度为20mg/kg的土壤中，每4d浇灌1次清水，每2d喷施1次1.28mg/L的BR，喷洒时加入体积分数为0.2％的吐温20作为展开剂，用喷雾器在植株叶片正反两面均匀喷施，以叶片正反两面全部湿润且无液体滴下，以保证喷施充分、均匀。When the tobacco seedlings grow to 5 leaves and 1 heart, select tobacco seedlings with good shape and consistent growth, move them into the soil with a concentration of cadmium chloride of 20mg/kg, water once every 4d, and spray 1.28mg/kg once every 2d. For BR of L, add Tween 20 with a volume fraction of 0.2% as a developer when spraying, and spray evenly on both sides of the leaves of the plant with a sprayer, so that both sides of the leaves are wet and there is no liquid dripping to ensure sufficient spraying, uniform.
步骤(3)同实施例4步骤(3)。Step (3) is the same as embodiment 4 step (3).
对照例4：Comparative example 4:
步骤(1)同实施例1步骤(1)；Step (1) is with embodiment 1 step (1);
(2)烟草幼苗的处理：(2) The processing of tobacco seedlings:
当烟草幼苗生长至5叶1心时，选取形态良好与长势一致的烟草幼苗，移入氯化镉浓度为20mg/kg的土壤中，每4d浇灌1次清水，每2d喷施1次1.28mg/L的SA，喷洒时加入体积分数为0.2％的吐温20作为展开剂，用喷雾器在植株叶片正反两面均匀喷施，以叶片正反两面全部湿润且无液体滴下，以保证喷施充分、均匀。When the tobacco seedlings grow to 5 leaves and 1 heart, select tobacco seedlings with good shape and consistent growth, move them into the soil with a concentration of cadmium chloride of 20mg/kg, water once every 4d, and spray 1.28mg/kg once every 2d. For SA of L, add Tween 20 with a volume fraction of 0.2% as a developer when spraying, and spray evenly on both sides of the leaves of the plant with a sprayer, so that both sides of the leaves are wet and there is no liquid dripping to ensure sufficient spraying, uniform.
步骤(3)同实施例4步骤(3)。Step (3) is the same as embodiment 4 step (3).
实验结果：Experimental results:
(1)从图1、2中可以看出，对照例2中烟草在受到Cd胁迫时体内的活性氧含量明显升高，与对照例2相比，对照例3、4烟草体内的活性氧含量显著降低，实施例4烟草体内的活性氧含量进一步降低，表明BR和SA的组合物(实施例1)能更高效地清除烟草体内产生的活性氧，从而提高对Cd的抗性。(1) As can be seen from Figures 1 and 2, the active oxygen content in the tobacco body in the control example 2 increased significantly when subjected to Cd stress. Compared with the control example 2, the active oxygen content in the tobacco body in the control examples 3 and 4 Significantly reduced, the active oxygen content in the tobacco body in Example 4 was further reduced, indicating that the composition of BR and SA (Example 1) can more efficiently remove the active oxygen generated in the tobacco body, thereby improving the resistance to Cd.
(2)由图3可知，对照例2烟草体内的丙二醛含量最高，说明Cd加剧了烟草细胞脂膜的过氧化程度,实施例4烟草体内的丙二醛含量最低，表明BR和SA的组合物能最大程度抑制膜脂过氧化，缓解Cd对烟草植株的氧化胁迫。(2) As can be seen from Fig. 3, the malondialdehyde content in the tobacco body of comparative example 2 is the highest, indicating that Cd has aggravated the degree of peroxidation of the tobacco cell lipid membrane, and the malondialdehyde content in the tobacco body of embodiment 4 is the lowest, showing that BR and SA The composition can inhibit membrane lipid peroxidation to the greatest extent and alleviate the oxidative stress of Cd on tobacco plants.
实验证明，实施例2、实施例3的组合物对烟草H2O2含量、O2 -含量和丙二醛含量的影响与实施例1的组合物相似。Experiments have shown that the effects of the compositions of Example 2 and Example 3 on tobacco H 2 O 2 content, O 2 - content and malondialdehyde content are similar to those of the composition of Example 1.
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