WO2024174288A1 - 一种利用硅层层自组装增强植物重金属铬抗性的方法 - Google Patents

一种利用硅层层自组装增强植物重金属铬抗性的方法 Download PDF

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WO2024174288A1
WO2024174288A1 PCT/CN2023/079938 CN2023079938W WO2024174288A1 WO 2024174288 A1 WO2024174288 A1 WO 2024174288A1 CN 2023079938 W CN2023079938 W CN 2023079938W WO 2024174288 A1 WO2024174288 A1 WO 2024174288A1
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silicon
layer
chromium
plant
cationic
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庞志豪
梁永超
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Zhejiang University
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Zhejiang University
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G7/00—Botany in general
    • A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
    • 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
    • A01G22/15—Leaf crops, e.g. lettuce or spinach 
    • 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
    • A01G22/20—Cereals
    • A01G22/22—Rice

Definitions

  • the present invention belongs to the technical field of agricultural environmental protection, and more specifically, relates to a method for enhancing the ability of plants to resist heavy metal chromium stress by utilizing silicon layer-by-layer self-assembly.
  • Chromium (Cr) is an important mineral resource and a highly toxic teratogenic and mutagenic heavy metal pollution element in the environment. It is listed as one of the primary carcinogens by the International Agency for Research on Cancer. Taking the Chinese Soil Environmental Quality Standard (GB15618-2018) as a reference, about 4.31% of the farmland sampling points in China have chromium exceeding the screening value, and 0.12% exceed the control value.
  • the simplest strategy to deal with heavy metal pollution in farmland is to prevent it from being absorbed by plants from the soil to prevent the metal from entering the aboveground part. How to maintain safe agricultural production in contaminated soil is an urgent problem to be solved.
  • Silicon is a beneficial element for plants. Since silicon can block metal ions from entering cells and inhibit the transport of metals from roots to aboveground parts after being deposited in cell walls, its role in helping plants resist metal stress has been widely reported. However, silicon polymerizes slowly in roots. In addition, due to the possible lack of transport proteins or certain specific components of cell walls, the silicon content of most dicotyledonous plants is significantly lower than that of monocotyledonous plants. Therefore, even if silicon is applied exogenously, the increase in silicon content in the plants is limited, and the effect of resisting adverse stress is not obvious. How to regulate the polymerization rate of plant silicon and how to enable non-silicon-loving plants to use silicon to increase their ability to resist adverse stress have become bottlenecks in plant silicon nutrition.
  • the purpose of the present invention is to quickly increase the silicon content of plant roots, enhance their resistance to heavy metal stress, and provide a safe, economical and practical operation scheme.
  • a method for enhancing plant resistance to heavy metal chromium by using silicon layer-by-layer self-assembly wherein the root system of the plant is alternately immersed in a cationic silicon polymerization inducer solution and a silicic acid solution before the plant is transplanted.
  • the layer-by-layer self-assembly method forms one or more layers of silicon on the surface of the plant roots, and then the plants are transplanted into a chromium-containing environment.
  • the plant is a monocotyledon or a dicotyledon; the monocotyledon is preferably rice; the dicotyledon is preferably pakchoi.
  • the cationic silicon polymerization inducer in the cationic silicon polymerization inducer solution is one or more of polyquaternium-10, cationic guar gum, gelatin, chitosan, poly-L-lysine, aqueous polyurethane, polyacrylamine hydrochloride, cationic polyacrylamide, polydimethyldiallylammonium chloride, and polyethyleneimine.
  • the cationic silicon polymerization inducer in the cationic silicon polymerization inducer solution is cationic guar gum.
  • the roots of the plants are alternately immersed in a cationic silicon polymerization inducer solution and a silicic acid solution for two rounds, so that two silicon layers are formed on the surface of the plant roots.
  • the root system of the plant is immersed in the cationic silicon polymerization inducer solution and the silicic acid solution for 10 minutes to 1 hour respectively.
  • the root system of the plant is immersed in the cationic silicon polymerization inducer solution and the silicic acid solution for 20 minutes respectively.
  • the concentration of the cationic silicon polymerization inducer solution is 0.5% by mass.
  • the concentration of the silicic acid solution is 100 mM.
  • the chromium-containing environment is chromium-containing soil or nutrient solution.
  • the present invention has the following beneficial effects: in a heavily chromium-polluted environment (chromium concentration of the nutrient solution is 500 ⁇ M), under normal culture conditions, the chromium content of the aboveground part of the rice increases significantly after 3 days of stress, the growth is significantly inhibited, and a large number of rice plants die.
  • chromium concentration of the nutrient solution 500 ⁇ M
  • the silicon polymerization on the root surface is slow, the silicon deposition is less, and the ability to alleviate chromium stress is weak.
  • the layer-by-layer self-assembly method of the present invention can form a silicon layer on the surface of the plant root system, and the silicon content of the root system can be increased to a maximum of 1733 ⁇ g/g in a short period of time, and the chromium content of the aboveground part is reduced by 68%-75.2% compared with ordinary culture.
  • the present invention is superior to directly adding silicon to the nutrient solution in terms of rice growth conditions, increasing the silicon content of the root system, and limiting the chromium content of the aboveground part.
  • FIG. 1 Surface morphology of rice roots after pretreatment with different materials.
  • Ck means soaking in water for 40 min only
  • P0si means soaking in water for 20 min and then in silicate solution for 20 min
  • P1-P10 means soaking in ten polymer materials. After soaking in the solution for 20 min, it was soaked in silicate for 20 min.
  • the root surface of Ck was smooth, the P0si treatment had fine particles, and the P1-P10 treatments had deposits of various morphologies;
  • Figure 2 Effects of ten materials on silicon content and chromium content of rice under chromium stress.
  • A represents silicon content
  • the bar graph in B represents chromium content
  • the dotted line graph represents the chromium transport coefficient.
  • P1-P10 represents soaking in ten polymer material solutions for 20 minutes, then soaking in silicic acid for 20 minutes, and then placed in 1/4 Hoagland nutrient solution containing 500 ⁇ M chromium
  • P0si represents soaking in water for 20 minutes, then soaking in silicic acid solution for 20 minutes, and then placed in chromium-containing nutrient solution
  • Cr represents soaking in water for 40 minutes, and then placed in chromium-containing nutrient solution
  • Ck represents soaking in water only for 40 minutes, and then placed in nutrient solution without chromium stress.
  • Data represent the mean and standard error of three replicates, and data with different letters are significantly different at p ⁇ 0.05;
  • Figure 3 Effects of different treatments on the silicon content and chromium content of rice under chromium stress. There are five immersion times of 10min, 20min, 30min, 40min, and 1h, and five self-assembly layers of 1, 2, 3, 4, and 5.
  • A represents the silicon content
  • the bar graph in B represents the chromium content
  • the dotted line graph represents the chromium transport coefficient.
  • the data represent the mean and standard error of three replicates, and the data with different letters are significantly different when p ⁇ 0.05;
  • Figure 4 Correlation analysis between root silicon content and root and aboveground chromium content in the method optimization experiment.
  • A represents the relationship between root silicon and root chromium content
  • B represents the relationship between root silicon and aboveground chromium content.
  • the 95% confidence interval is also shown;
  • Figure 5 Effects of optimal materials and methods on silicon content and chromium content of pakchoi under chromium stress.
  • A represents silicon content
  • the bar graph in B represents chromium content
  • the dotted line graph represents the chromium transport coefficient.
  • G represents the use of the method of the present invention under 500 ⁇ M chromium stress
  • Si represents the direct addition of silicon to the chromium nutrient solution
  • Ck represents 500 ⁇ M chromium stress.
  • the data represent the mean and standard error of three replicates, and data with different letters are significantly different when p ⁇ 0.05. This method significantly increases the silicon content in the roots, reduces the chromium content in the aboveground part, and reduces the transport coefficient, and its effect is better than directly adding silicon to the nutrient solution.
  • the present invention provides a method for enhancing the heavy metal chromium resistance of plants by utilizing layer-by-layer self-assembly of silicon.
  • the method can quickly increase the silicon content in the roots of plants and improve their resistance to heavy metal chromium stress.
  • the specific method is that before transplanting the plants, the roots of the plants are alternately immersed in a cationic silicon polymerization inducer solution and a silicic acid solution, and one or more layers of silicon are formed on the surface of the plant roots by a layer-by-layer self-assembly method, and then the plants are transplanted into a chromium-containing environment.
  • the present invention can be applied to monocotyledonous plants (such as rice) or dicotyledonous plants (such as Chinese cabbage).
  • monocotyledonous plants such as rice
  • dicotyledonous plants such as Chinese cabbage.
  • a layer of silicon can be formed on the surface of the rice root system by alternately soaking in a cationic silicon polymerization inducer solution and a silicic acid solution.
  • the cationic silicon polymerization inducer solution is a solution prepared by cationic polymers.
  • the rice root system is soaked in the cationic silicon polymerization inducer solution to form a cationic polymer layer on the root system surface, and then the root system is placed in a silicic acid solution. Since the positive charge of the cationic polymer can promote the polymerization of silicic acid, a layer of silicon can be formed on the root surface. If it is necessary to continue to form a second layer of silicon, the root system needs to be soaked in the cationic polymer solution again, and then transferred to the silicic acid, and then the second layer of silicon is formed on the surface of the first layer of silicon.
  • the number of silicon layers on the root system surface can be adjusted.
  • the method can quickly increase the silicon content in the rice root, reduce the chromium content in the aboveground part, reduce the chromium transport coefficient, and improve the growth of rice under chromium stress.
  • the layer-by-layer self-assembly method of the present invention can also be applied to dicotyledonous plants represented by Chinese cabbage, increase the silicon content in the Chinese cabbage root, and significantly reduce the chromium content in the aboveground part.
  • the cationic silicon polymerization inducer in the cationic silicon polymerization inducer solution of the present invention can be a cationic polymer material, which can be a natural cationic polymer or a synthetic organic polymer.
  • the natural cationic polymers that can be used include: 1. polyquaternium salt-10 (cationic cellulose), 2. cationic guar gum, 3. gelatin, 4. chitosan, 5. poly-L-lysine, and the synthetic organic polymers that can be used include: 6. water-based polyurethane, 7. polyacrylamine hydrochloride, 8. cationic polyacrylamide, 9. polydimethyldiallylammonium chloride, and 10. polyethyleneimine.
  • cationic guar gum is recommended as the cationic polymer for inducing rapid silicon polymerization. It is non-toxic, widely available, and inexpensive, and can effectively promote silicon polymerization, and the effect of improving the chromium resistance of rice is very obvious.
  • the anti-chromium stress effects of different immersion times and numbers of silicon self-assembly layers were screened, and the recommended immersion time was 20 minutes, and two rounds of immersion were the best options.
  • the silicon content in the rice roots increased rapidly, the chromium content in the aboveground part was significantly reduced, and the growth condition was good.
  • the plants processed by the layer-by-layer self-assembly of the present invention can be transplanted into a chromium-containing environment such as chromium-containing soil or nutrient solution to alleviate chromium stress.
  • the method of enhancing plant heavy metal chromium resistance by using silicon layer-by-layer self-assembly includes the following steps:
  • ten cationic polymer materials are selected as cationic silicon polymerization inducers, namely: 1. Polyquaternium-10 (cationic cellulose), 2. Cationic guar gum, 3. Gelatin, 4. Chitosan, 5. Poly-L-lysine, 6. Water-based polyurethane, 7. Polyacrylamine hydrochloride, 8. Cationic polyacrylamide, 9. Polydimethyldiallyl ammonium chloride, 10. Polyethyleneimine.
  • the above ten cationic polymer materials are dissolved in deionized water and prepared into solutions with a mass fraction of 0.5% for standby use.
  • Rice seedlings (variety Nipponbare) with consistent growth conditions 21 days after germination were selected for the experiment.
  • the specific method is: soak the roots in a cationic silicon polymerization inducer solution for 20 minutes to form a polymer layer on its surface, and shake it continuously during the period. Then soak the roots in a 100mM silicic acid solution for 20 minutes. Since the positive charge of cationic polymers can promote the polymerization of silicic acid, a silicon layer can be formed.
  • Ten cationic silicon polymerization inducer solutions are each set up as a treatment group, recorded as P1-P10.
  • the rice seedlings with different treatments were placed in a 1/4 Hoagland nutrient solution containing 500 ⁇ M Cr(III) (prepared by chromium chloride) and cultured for 3 days.
  • CK CK
  • P0si chromium-free 1/4 Hoagland nutrient solution for 3 days.
  • P0si the rice seedlings were immersed in pure water for 20 minutes, and then immersed in a silicate solution for 20 minutes, and then placed in 1/4 Hoagland nutrient solution containing 500 ⁇ M Cr(III) (prepared by chromium chloride) for 3 days.
  • the rice seedlings were immersed in 1/4 Hoagland nutrient solution with 1mM silicate added for 40 minutes, and then placed in 1/4 Hoagland nutrient solution containing 500 ⁇ M Cr(III) (prepared by chromium chloride).
  • the Cr group was cultured for 3 days.
  • the rice seedlings were immersed in pure water for 40 minutes and then placed in 1/4 Hoagland nutrient solution containing 500 ⁇ M Cr(III) (prepared with chromium chloride).
  • the pH of the nutrient solution in different treatment groups was adjusted to 5.6 to keep it consistent among treatments.
  • the silicon deposition on the surface of rice roots where the rice seedlings have completed immersion but have not yet been transferred to the nutrient solution is shown in Figure 1.
  • the root surface of the Ck group is smooth
  • the P0si group has fine particles
  • the P1-P10 treatment groups have deposition in various forms.
  • the silicon and chromium contents of the shoots and roots measured after 3 days of cultivation are shown in Figure 2.
  • the P2 (cationic guar gum) treatment group had the best effect, with the silicon content in the roots being 2.49 times that of the silicic acid soaking treatment group, the chromium content in the shoots being 75.2% lower than that of the Cr treatment group, and the chromium transport coefficient being reduced to 0.06.
  • the thickness of the silicon layer of the root cell wall can be controlled by adjusting the immersion time and the number of alternating adsorption of cationic polymers and silicate.
  • a cationic silicon polymerization inducer solution with a mass fraction of 0.5% was prepared with cationic guar gum, and rice seedlings (21 days after germination, the variety was Nipponbare) were alternately soaked in the cationic silicon polymerization inducer solution and 100mM silicic acid solution once.
  • this embodiment set five soaking times of 10min, 20min, 30min, 40min, and 1h, which were recorded as T10 group, T20 group, T30 group, T40 group, and T60 group respectively.
  • the soaking time here refers to the soaking time in each of the two solutions.
  • the soaking time of 10min means soaking in the cationic silicon polymerization inducer solution for 10min and then soaking in the 100mM silicic acid solution for 10min.
  • the immersion time was kept unchanged at 20 min in this embodiment, and the alternating immersion times in the cationic silicon polymerization inducer solution and the 100 mM silicic acid solution were set to be 1, 2, 3, 4, and 5 times, respectively, which were recorded as Group 1, Group 2, Group 3, Group 4, and Group 5.
  • the rice seedlings with different treatments were placed in a 1/4 Hoagland nutrient solution containing 500 ⁇ M Cr (III) (prepared by chromium chloride), cultured for 3 days, and then taken out to measure the silicon and chromium content of the aboveground part (Shoots) and the underground part (Roots).
  • 500 ⁇ M Cr (III) prepared by chromium chloride
  • the same CK group and Cr group as in Example 1 were set in this embodiment.
  • the method of enhancing plant resistance to heavy metal chromium by layer-by-layer self-assembly of silicon is applied to dicotyledonous plants (Brachys pakchoi).
  • the optimal materials, immersion time and number of layers (cationic guar gum, 20 min, 2 rounds) determined by the above embodiment experiments were used to treat the root system of Brassica chinensis.
  • a cationic silicon polymerization inducer solution with a mass fraction of 0.5% was prepared with cationic guar gum, and the roots of Brassica chinensis were alternately immersed in the cationic silicon polymerization inducer solution and 100 mM silicic acid solution for 2 times, with the immersion time in each solution being 20 min.
  • the roots of Brassica chinensis were then placed in a 1/4 Hoagland nutrient solution containing 500 ⁇ M Cr (III) (prepared by chromium chloride) and cultured for 3 days.
  • this embodiment also sets up a deionized water soaking group CK for the same time, and a treatment group Si in which 1mM silicic acid is added to the nutrient solution.
  • the method of the CK group is to soak the roots of the Chinese cabbage in pure water for 40 minutes, and then place them in 1/4 Hoagland nutrient solution containing 500 ⁇ M Cr(III) (prepared by chromium chloride) for 3 days.
  • the specific method of the Si group is to soak the roots of the Chinese cabbage in 1/4 Hoagland nutrient solution with 1mM silicic acid added for 40 minutes, and then place the roots of the Chinese cabbage in 1/4 Hoagland nutrient solution containing 500 ⁇ M Cr(III) (prepared by chromium chloride) for 3 days.

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Abstract

本发明公开了一种利用硅层层自组装增强植物重金属铬抗性的方法。该方法具体为：在植物移栽之前，将植物的根系在阳离子硅聚合诱导剂溶液和硅酸溶液中交替浸泡，通过层层自组装的方法在植物根系表面形成一层或多层硅，然后再将植物移栽至含铬环境中。使用本发明的方法可在植物根系表面形成硅层，使根系硅含量在短时间内可最高增加至1733μg/g，与普通培养相比地上部铬含量降低68％-75.2％。此外，本发明在水稻生长状况、增加根系硅含量、限制地上部铬含量等方面都优于营养液直接加硅。

Description

一种利用硅层层自组装增强植物重金属铬抗性的方法 技术领域
本发明属于农业环保技术领域，更具体地，本发明涉及一种利用硅层层自组装增强植物抗重金属铬胁迫能力的方法。
背景技术
随着工业活动的进行，造成部分农田的重金属污染，这会损害植物生长发育，造成粮食大量减产。此外，重金属在植物中积累并在食物链中富集，还将威胁动物和人类健康。铬(Cr)是重要的矿产资源，也是环境中毒性较大的致畸、致突变重金属污染元素，被国际癌症研究机构列为首要致癌物之一。以中国土壤环境质量标准(GB15618-2018)作为参考，中国约4.31％的农田采样点铬超过筛查值，0.12％的超过控制值。应对农田重金属污染最简单的策略是避免其从土壤中被植物吸收，以防止金属进入地上部。如何在污染土壤中维持农业安全生产，是亟待解决的问题。
硅是一种植物有益元素。由于硅其在细胞壁沉积后可以阻滞金属离子进入细胞，抑制金属从根向地上部的转运，其在帮助植物抵抗金属胁迫中的作用已经被广泛报道。但硅在根部聚合速度较慢。此外，由于可能缺乏转运蛋白或细胞壁某些特定组分，大部分双子叶植物硅含量较单子叶植物大幅降低，因此即使外源施硅，其植株硅含量提高幅度有限，抗逆境胁迫效果也并不明显。如何调控植物硅聚合速度，如何让非喜硅植物也可以利用硅增加抗逆境胁迫能力，这两大问题成为植物硅营养的瓶颈。
发明内容
本发明的目的在于快速提高植物根部硅含量，增强其抗重金属胁迫能力，并提供一种安全、经济、实用的操作方案。
为实现上述目的，本发明所采用的具体技术方案如下：
一种利用硅层层自组装增强植物重金属铬抗性的方法，其做法为：在植物移栽之前，将植物的根系在阳离子硅聚合诱导剂溶液和硅酸溶液中交替浸泡，通过 层层自组装的方法在植物根系表面形成一层或多层硅，然后再将植物移栽至含铬环境中。
作为优选，所述植物为单子叶植物或双子叶植物；所述单子叶植物优选为水稻；所述双子叶植物优选为小白菜。
作为优选，所述阳离子硅聚合诱导剂溶液中的阳离子硅聚合诱导剂为聚季铵盐-10、阳离子瓜尔胶、明胶、壳聚糖、多聚左旋赖氨酸、水性聚氨酯、聚丙烯胺盐酸盐、阳离子型聚丙烯酰胺、聚二甲基二烯丙基氯化铵、聚乙烯亚胺中的一种或多种。
作为优选，所述阳离子硅聚合诱导剂溶液中的阳离子硅聚合诱导剂为阳离子瓜尔胶。
作为优选，所述植物的根系在阳离子硅聚合诱导剂溶液和硅酸溶液中交替浸泡两轮，在植物根系表面形成两层硅层。
作为优选，所述植物的根系在阳离子硅聚合诱导剂溶液和硅酸溶液中各自的浸泡时长为10min～1h。
作为优选，所述植物的根系在阳离子硅聚合诱导剂溶液和硅酸溶液中各自的浸泡时长均为20min。
作为优选，所述阳离子硅聚合诱导剂溶液的浓度为质量分数0.5％。
作为优选，所述硅酸溶液的浓度为100mM。
作为优选，所述含铬环境为含铬的土壤或营养液。
本发明与现有技术相比具有如下有益效果：在重度铬污染环境中(营养液铬浓度为500μM)，普通培养条件下，胁迫3天水稻地上部铬含量大幅增加，生长受到显著抑制，大量死亡。而将根系短时间浸泡在硅酸中，根系表面硅聚合缓慢，硅沉积较少，缓解铬胁迫能力较弱。使用本发明的层层自组装方法可在植物根系表面形成硅层，根系硅含量在短时间内可最高增加至1733μg/g，与普通培养相比地上部铬含量降低68％-75.2％。此外，本发明在水稻生长状况、增加根系硅含量、限制地上部铬含量等方面都优于营养液直接加硅。
附图说明
图1不同材料预处理后水稻根表形貌。Ck表示只在水中浸泡40min，P0si表示在水中浸泡20min后在硅酸溶液中浸泡20min，P1-P10代表在十种高分子材 料溶液中浸泡20min后在硅酸中浸泡20min。在500nm比例尺下，Ck根表光滑，P0si处理有细小颗粒，而P1-P10处理有多种形态的沉积；
图2十种材料对铬胁迫下水稻硅含量和铬含量的影响。A代表硅含量，B中柱状图代表铬含量，点线图表示铬转运系数。P1-P10代表在十种高分子材料溶液中浸泡20min后在硅酸中浸泡20min，而后置于含500μM铬的1/4霍格兰营养液中，P0si表示在水中浸泡20min后在硅酸溶液中浸泡20min，而后置于含铬营养液中，Cr表示在水中浸泡40min，而后置于含铬营养液中，Ck表示只在水中浸泡40min，而后置于无铬胁迫的营养液中。数据代表三个重复的平均值和标准误，不同字母的数据在p<0.05时显著不同；
图3不同处理方法对铬胁迫下水稻硅含量和铬含量的影响。设有设置10min、20min、30min、40min、1h五个浸泡时间，1、2、3、4、5五个自组装层数。A代表硅含量，B中柱状图代表铬含量，点线图表示铬转运系数。数据代表三个重复的平均值和标准误，不同字母的数据在p<0.05时显著不同；
图4方法优化实验中根系硅含量与根系和地上部铬含量的相关性分析。A代表根系硅与根系铬含量的关系，B代表根系硅与地上部铬含量的关系。同时显示95％置信区间；
图5最优材料和方法对铬胁迫下小白菜硅含量和铬含量的影响。A代表硅含量，B中柱状图代表铬含量，点线图表示铬转运系数。G代表500μM铬胁迫下使用本发明的方法，Si表示铬营养液直接加硅，Ck表示500μM铬胁迫。数据代表三个重复的平均值和标准误，不同字母的数据在p<0.05时显著不同。本方法显著增加根部硅含量，降低地上部铬含量，降低转运系数，其效果优于营养液直接加硅。
具体实施方式
为使本发明的上述目的、特征和优点能够更加明显易懂，下面结合附图对本发明的具体实施方式做详细的说明。在下面的描述中阐述了很多具体细节以便于充分理解本发明。但是本发明能够以很多不同于在此描述的其它方式来实施，本领域技术人员可以在不违背本发明内涵的情况下做类似改进，因此本发明不受下面公开的具体实施例的限制。本发明各个实施例中的技术特征在没有相互冲突的前提下，均可进行相应组合。
因此本发明提供了一种利用硅层层自组装增强植物重金属铬抗性的方法，该方法可快速增加植物根部硅含量并提高其抗重金属铬胁迫能力的方法，其具体做法是在植物移栽之前，将植物的根系在阳离子硅聚合诱导剂溶液和硅酸溶液中交替浸泡，通过层层自组装的方法在植物根系表面形成一层或多层硅，然后再将植物移栽至含铬环境中。
本发明可应用于单子叶植物(如水稻)或双子叶植物(如小白菜)中。以水稻为例，由于水稻根系细胞壁表面呈负电性，在上述层层自组装方法中，在阳离子硅聚合诱导剂溶液和硅酸溶液中交替浸泡一轮，可以在水稻根系表面形成一层硅。阳离子硅聚合诱导剂溶液是由阳离子高分子配制而成的溶液。在这个交替浸泡的过程中，水稻根系浸泡在阳离子硅聚合诱导剂溶液中可以在根系表面形成阳离子高分子层，而后将根系置于硅酸溶液中，由于阳离子高分子的正电性可以促进硅酸聚合，因此可以在根表形成一层硅层。如果需要继续形成第二层硅，则需再将根系置于阳离子高分子溶液中浸泡后，转移至硅酸中，进而在第一层硅表面继续形成第二层硅。由此，通过控制水稻根系在阳离子硅聚合诱导剂溶液和硅酸溶液中交替浸泡的轮数，可以调节根系表面硅的层数。该方法能够快速增加水稻根部硅含量，降低地上部铬含量，降低铬转运系数，并改善铬胁迫下水稻生长状况。同样的，本发明的层层自组装方法也可以应用于以小白菜为代表的双子叶植物上，增加小白菜根部硅含量，显著降低其地上部铬含量。
本发明中阳离子硅聚合诱导剂溶液中的阳离子硅聚合诱导剂，可以采用阳离子高分子材料，阳离子高分子材料可以是天然阳离子高分子，也可以是合成有机高分子，可采用的天然阳离子高分子包括：1.聚季铵盐-10(阳离子纤维素)、2.阳离子瓜尔胶、3.明胶、4.壳聚糖、5.多聚左旋赖氨酸，可采用的合成有机高分子包括：6.水性聚氨酯、7.聚丙烯胺盐酸盐、8.阳离子型聚丙烯酰胺、9.聚二甲基二烯丙基氯化铵、10.聚乙烯亚胺。综合考量安全性、产品价格和作用效果，诱导硅快速聚合的阳离子高分子推荐使用阳离子型瓜尔胶，其无毒害，来源广泛，且价格低廉，能够有效促进硅聚合，提高水稻抗铬能力的效果十分明显。
另外，对不同浸泡时间和硅自组装层数的抗铬胁迫效果进行筛选，推荐浸泡时间20min，浸泡两轮为最佳方案，水稻根系硅含量迅速增加，地上部铬含量明显降低，生长状况良好。
另外，由于植物以单硅酸形式吸硅，推荐使用100mM硅酸，可由硅酸钠酸化制备或通过氢型阳离子交换树脂制备。阳离子高分子推荐在使用前溶于去离子水，配置成质量分数0.5％的溶液。
本发明经过层层自组装处理后的植物,可移栽至含铬的土壤或营养液等含铬环境中，缓解铬胁迫。
下面通过若干实施例对本发明的具体实现以及技术效果进行详细描述。
实施例1
本实施例中，利用硅层层自组装增强植物重金属铬抗性的方法，包括如下步骤：
(1)配制阳离子硅聚合诱导剂溶液和硅酸溶液
本实施例中，选择十种阳离子高分子材料作为阳离子硅聚合诱导剂，分别为：1.聚季铵盐-10(阳离子纤维素)、2.阳离子瓜尔胶、3.明胶、4.壳聚糖、5.多聚左旋赖氨酸、6.水性聚氨酯、7.聚丙烯胺盐酸盐、8.阳离子型聚丙烯酰胺、9.聚二甲基二烯丙基氯化铵、10.聚乙烯亚胺。将上述十种阳离子高分子材料溶于去离子水，分别配制成质量分数0.5％的溶液待用。
另外，配制100mM硅酸溶液待用。
(2)不同处理组实验
选择发芽后21d，生长状况一致的水稻幼苗(品种为日本晴)进行实验。具体做法为：将根系浸泡在阳离子硅聚合诱导剂溶液中20min，使其表面形成高分子层，期间不断摇晃。而后将根系浸泡于100mM硅酸溶液中20min，由于阳离子高分子的正电性可以促进硅酸聚合，可以形成一层硅层。十种阳离子硅聚合诱导剂溶液分别设置一个处理组，记为P1-P10。将不同处理的水稻幼苗置于含有500μM Cr(Ⅲ)(由氯化铬配置)的1/4霍格兰营养液中，培养3d。
另外，设置四个对比处理组CK、P0si、Si和Cr。其中CK组的做法为将水稻幼苗在纯水中浸泡40min后，置于无铬的1/4霍格兰营养液中，培养3d。P0si组的做法为将水稻幼苗浸泡在纯水中20min后再在硅酸溶液中浸泡20min，再将水稻幼苗置于含有500μM Cr(Ⅲ)(由氯化铬配置)的1/4霍格兰营养液中，培养3d。Si组的做法为将水稻幼苗浸泡在加入1mM硅酸的1/4霍格兰营养液中40min，再将水稻幼苗置于含有500μM Cr(Ⅲ)(由氯化铬配置)的1/4霍格兰营养液中， 培养3d。Cr组的做法为将水稻幼苗浸泡在纯水中40min，再将水稻幼苗置于含有500μM Cr(Ⅲ)(由氯化铬配置)的1/4霍格兰营养液中。
本实施例中，不同处理组的营养液pH均调至5.6，以保持处理间一致。
对于上述P1-P10以及CK、P0si这12个处理组，水稻幼苗完成浸泡但尚未转移至营养液的水稻根系表面硅沉积情况如图1所示。从图中可见，Ck组根表光滑，P0si组处理有细小颗粒，而P1-P10处理组有多种形态的沉积。
培养3d后测定的地上部(Shoots)和地下部(Roots)硅、铬含量如图2。从图中可见，P2(阳离子型瓜尔胶)处理组效果最好，根部硅含量是硅酸浸泡处理组的2.49倍，地上部铬含量较Cr处理组降低75.2％，铬转运系数降至0.06。
实施例2
本实施例中，通过调节不同参数进行实验，来确定利用硅层层自组装增强植物重金属铬抗性的方法的最佳浸泡时间和最佳层数。根系细胞壁硅层的厚度可以通过调节浸泡时间和交替吸附阳离子高分子和硅酸的次数进行控制。
以阳离子瓜尔胶配制质量分数0.5％的阳离子硅聚合诱导剂溶液，将水稻幼苗(发芽后21d，品种为日本晴)在阳离子硅聚合诱导剂溶液和100mM硅酸溶液中交替浸泡1次，为了对比不同浸泡时间对最终结果的影响，本实施例设置了10min、20min、30min、40min、1h五个浸泡时间，分别记为T10组、T20组、T30组、T40组、T60组。此处的浸泡时间是指在两种溶液中各自浸泡的时间，例如浸泡时间10min是指在阳离子硅聚合诱导剂溶液中浸泡10min，再在100mM硅酸溶液中浸泡10min。另外，为了对比层层自组装过程中硅层的层数对最终结果的影响，本实施例中保持浸泡时间20min不变，设置在阳离子硅聚合诱导剂溶液和100mM硅酸溶液中交替浸泡次数分别为1、2、3、4、5次，分别记为1组、2组、3组、4组、5组。然后再将上述不同处理的水稻幼苗置于含有500μM Cr(Ⅲ)(由氯化铬配置)的1/4霍格兰营养液中，培养3d，再取出测定地上部(Shoots)和地下部(Roots)的硅、铬含量。另外，本实施例中设置了与实施例1相同的CK组和Cr组。
本实施例的结果如图3，最终确定浸泡20min、交替2次为本发明层层自组装过程的最佳参数。另外，对本实施例实验中根部硅含量与地上部铬含量进行线性拟合分析，图4显示了本实施例实验中根部硅含量与地上部铬含量的显著负相 关关系。
实施例3
本实施例中，将利用硅层层自组装增强植物重金属铬抗性的方法应用于双子叶植物(小白菜)上。利用上述实施例实验确定的最佳材料、浸泡时间和层数(阳离子瓜尔胶、20min、2轮)，处理小白菜根系。处理过程中，以阳离子瓜尔胶配制质量分数0.5％的阳离子硅聚合诱导剂溶液，将小白菜根系在阳离子硅聚合诱导剂溶液和100mM硅酸溶液中交替浸泡2次，每种溶液中浸泡时间为20min。然后将小白菜根系置于含有500μM Cr(Ⅲ)(由氯化铬配置)的1/4霍格兰营养液中，培养3d。
同时，本实施例还设置了相同时间的去离子水浸泡组CK，以及营养液添加1mM硅酸的处理组Si。其中CK组的做法为将小白菜根系在纯水中浸泡40min后，置于含有500μM Cr(Ⅲ)(由氯化铬配置)的1/4霍格兰营养液中，培养3d。Si组的具体做法将小白菜根系浸泡在加入1mM硅酸的1/4霍格兰营养液中40min，再将小白菜根系置于含有500μM Cr(Ⅲ)(由氯化铬配置)的1/4霍格兰营养液中，培养3d。
上述三个处理组培养3d后测定的地上部(Shoots)和地下部(Roots)硅、铬含量结果如图5，结果表明使用本发明方法后白菜根部硅含量显著提高，高于营养液加硅，达1502μg/g，与在水稻上使用该方法后的硅含量接近。而地上部硅含量增加不如营养液加硅组。使用本发明方法后，地上部铬含量显著降低，从75.8μg/g降至19.0μg/g，转移系数从0.027降低至0.009。
通过上述的说明内容相关工作人员完全可以在不偏离本项发明技术思想的范围内，进行多样的变更以及修改。本项发明的技术性范围并不局限于说明书上的内容，必须要根据权利要求范围来确定其技术性范围。以上所述的实施例只是本发明的一种较优的方案，然其并非用以限制本发明。有关技术领域的普通技术人员，在不脱离本发明的精神和范围的情况下，还可以做出各种变化和变型。因此凡采取等同替换或等效变换的方式所获得的技术方案，均落在本发明的保护范围内。

Claims (10)

  1. 一种利用硅层层自组装增强植物重金属铬抗性的方法，其特征在于，在植物移栽之前，将植物的根系在阳离子硅聚合诱导剂溶液和硅酸溶液中交替浸泡，通过层层自组装的方法在植物根系表面形成一层或多层硅，然后再将植物移栽至含铬环境中。
  2. 如权利要求1所述的利用硅层层自组装增强植物重金属铬抗性的方法，其特征在于，所述植物为单子叶植物或双子叶植物；所述单子叶植物优选为水稻；所述双子叶植物优选为小白菜。
  3. 如权利要求1所述的利用硅层层自组装增强植物重金属铬抗性的方法，其特征在于，所述阳离子硅聚合诱导剂溶液中的阳离子硅聚合诱导剂为聚季铵盐-10、阳离子瓜尔胶、明胶、壳聚糖、多聚左旋赖氨酸、水性聚氨酯、聚丙烯胺盐酸盐、阳离子型聚丙烯酰胺、聚二甲基二烯丙基氯化铵、聚乙烯亚胺中的一种或多种。
  4. 如权利要求1所述的利用硅层层自组装增强植物重金属铬抗性的方法，其特征在于，所述阳离子硅聚合诱导剂溶液中的阳离子硅聚合诱导剂为阳离子瓜尔胶。
  5. 如权利要求1所述的利用硅层层自组装增强植物重金属铬抗性的方法，其特征在于，所述植物的根系在阳离子硅聚合诱导剂溶液和硅酸溶液中交替浸泡两轮，在植物根系表面形成两层硅层。
  6. 如权利要求1所述的利用硅层层自组装增强植物重金属铬抗性的方法，其特征在于，所述植物的根系在阳离子硅聚合诱导剂溶液和硅酸溶液中各自的浸泡时长为10min～1h。
  7. 如权利要求1所述的利用硅层层自组装增强植物重金属铬抗性的方法，其特征在于，所述植物的根系在阳离子硅聚合诱导剂溶液和硅酸溶液中各自的浸泡时长均为20min。
  8. 如权利要求1所述的利用硅层层自组装增强植物重金属铬抗性的方法，其特征在于，所述阳离子硅聚合诱导剂溶液的浓度为质量分数0.5％。
  9. 如权利要求1所述的利用硅层层自组装增强植物重金属铬抗性的方法，其特征在于，所述硅酸溶液的浓度为100mM。
  10. 如权利要求1所述的利用硅层层自组装增强植物重金属铬抗性的方法，其特征在于，所述含铬环境为含铬的土壤或营养液。
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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103789114A (zh) * 2013-12-26 2014-05-14 广东省生态环境与土壤研究所 一种可以抑制水稻重金属吸收积累生产富硒稻米的硒掺杂纳米硅溶胶及其制备方法
CN108040844A (zh) * 2017-11-15 2018-05-18 佛山科学技术学院 一种抗铝毒豌豆幼苗培植方法
CN109729963A (zh) * 2019-02-01 2019-05-10 佛山科学技术学院 一种基于水稻材料的纳米硅沉积阻隔重金属方法
CN110679255A (zh) * 2019-10-16 2020-01-14 常州大学 一种降低水稻籽粒铬含量的稻田施硅方法
CN115024173A (zh) * 2022-06-17 2022-09-09 浙江理工大学 5-氨基乙酰丙酸和九水偏硅酸钠的组合在提高重金属镉胁迫下丹参有效成分中的应用
WO2022246880A1 (zh) * 2021-05-24 2022-12-01 中国科学院南京土壤研究所 一种降低稻米镉含量的轻简化方法及应用

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115365291A (zh) * 2022-08-26 2022-11-22 桂林理工大学 一种用于提高植物对镉富集能力的方法

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103789114A (zh) * 2013-12-26 2014-05-14 广东省生态环境与土壤研究所 一种可以抑制水稻重金属吸收积累生产富硒稻米的硒掺杂纳米硅溶胶及其制备方法
CN108040844A (zh) * 2017-11-15 2018-05-18 佛山科学技术学院 一种抗铝毒豌豆幼苗培植方法
CN109729963A (zh) * 2019-02-01 2019-05-10 佛山科学技术学院 一种基于水稻材料的纳米硅沉积阻隔重金属方法
CN110679255A (zh) * 2019-10-16 2020-01-14 常州大学 一种降低水稻籽粒铬含量的稻田施硅方法
WO2022246880A1 (zh) * 2021-05-24 2022-12-01 中国科学院南京土壤研究所 一种降低稻米镉含量的轻简化方法及应用
CN115024173A (zh) * 2022-06-17 2022-09-09 浙江理工大学 5-氨基乙酰丙酸和九水偏硅酸钠的组合在提高重金属镉胁迫下丹参有效成分中的应用

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