NO对蒜苔衰老的延缓作用及其机理的研究
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摘要
NO作为一种新型的细胞间和细胞内信息传递的信号分子成为人们研究的热点。已经发现NO对多种果蔬有延缓衰老的作用。蒜苔采后由于物质再转运导致食用品质下降,对于此类模式果蔬的衰老机制至今尚不完全清除。本研究以物质再转运的典型模式材料蒜苔为研究对象,研究一氧化氮(NO)对蒜苔衰老的延缓作用机制,为解决此类园艺产品的贮藏保鲜问题提供科学依据。
本实验利用不同浓度的一氧化氮(nitric oxide,NO)供体硝普钠(sodium nitro prusside,SNP)及NO和H_2O_2清除剂抑制剂处理,采朋生理生化方法结合激光共聚焦扫描显微镜技术,初步探索NO对蒜苔采后物质再转运作用机制。
实验结果表明:
1.适宜浓度SNP处理对蒜苔的保鲜效果显著。能够延缓蒜苔苔苞膨大、基部可溶性糖、可溶性蛋白含量的下降以及纤维化程度的升高,且苔茎保持鲜绿。其中,0.1mmol·L~(-1)的SNP浸泡苔茎基部3h效果最佳。高于0.5 mmol·L~(-1)的SNP处理,对蒜苔造成伤害。
2.SNP处理能显著地抑制苔苞直径的膨大及H_2O_2含量的增加。在4±1℃冷藏过程中,0.1mmol·L~(-1)的SNP处理抑制了苔苞膨大、苔茎基部H_2O_2含量增加、相对膜透性上升和POD活性的提高;提高了SOD、CAT和抗坏血酸过氧化物酶(APX)和顶部POD活性。
3.APase酶活性在贮藏前期有很高的活性,但随着贮藏时间的推移,活性呈下降趋势;SNP处理能够延缓APase酶活性高峰期的出现。
4.蒜苔中内源NO分布和内含物转运的程度有关:转运程度比较低的组织中分布比较均匀,随着转运程度的升高,逐步会由周缘维管束向中央维管束转移,但是,顶端组织NO分布和水平没有太大变化;外源NO处理能够诱导内源NO水平升高,这可能是通过NOS途径起作用。
5.H_2O_2参与蒜苔的物质转运过程。在整个转运过程中,H_2O_2在所有组织中都有分布,衰老组织中H_2O_2含量很少或者没有。生长旺盛顶端组织中,水平很高;高浓度的H_2O_2促进物质转运,降低基部H_2O_2含量,可以抑制物质转运。并且在物质转运过程中二者相互影响。
6.在蒜苔物质再转运过程中NO抑制物质转运的作用与内源H_2O_2的水平有关。
NO as a new type of intercellular and intracellar signaling molecule messaging become a research hotspot. NO has been found the role of delaying in a variety of fruits and vegetables senescence. Redistribution of cell in excised garlic scape lead to decline in eating quality, of such models of the aging mechanism of fruits and vegetables is unknown. In this study, garlic scape as the typical models of transport of substances, study the effects of NO on the role of senescence mechanisms transduction in excised garlic scape, in order to provide scientific basis to solve this kind of horticultural products of storing and keeping fresh problems.
Using sodium nitro prusside (SNP) as a nitric oxide (NO) donor, Hemoglobin (HB) as a NO scavenger, N~ω-nitro-L-arsinine methylter (L-NAME) as a NOS inhibitor, Sodium Tungstate as a NR inhibitor, Diphenyl iodine (DPI) as a NADPH oxidase inhibitor, Ascorbic acid (ASA) as a H_2O_2 scavenger, Physiological and biochemical methods combined with confocal laser scanning microscopy technology, Preliminary exploration of mechanisms of NO on transport of substances in excised garlic scape. The results as follows:
1. The results showed that suitable concentration of SNP had a remarkable effect on garlic scape preservation. It can inhibited the diameter of garlic bulbet and the degree of fibrosis increased, delayed the content of soluble suger and soluble protein in the basal of garlic scape, and the garlic scape were keeped viridity. 0.1 mmol·L~(-1) SNP solutions three hours at the basal of garlic scape was the best effect concentration in Preservation. But overhigh concentration of SNP (0.5mmol·L~(-1)) will cause the garlic sacape tissues injuries.
2. SNP treatment could significantly inhibit the increased of the diameter of garlic bulbet and the content of H_2O_2. 0.1 mmol·L~(-1) SNP inhibited the increased of the diameter of garlic bulbet and the content of H_2O_2 in basal garlic scape, relative membrane permeability increased, activities of peroxidase (POD) in basal garlic scape increased and promoted the activity of catalysis of superoxide dismutase (SOD), catalysis of catslase (CAT), ascorbate peroxidase (APX) in garlic scape and POD in apical garlic scape during four centigrade degree cold storage.
3. The acid phosphatase activities in the clove and the basal scape were very high, especially in the earier period of the pre-storage. However, with the time of torage, the acid phosphatase activities were a downward trend; Exogenous NO obviously slowed down the peak of acid phosphatase activities duing storage.
4. Distribution of endogenous NO related to the speed and extent of redistribution of cell content. Endogenous NO distributed evenly in low degree of transport of tissues, with the degree of transport increasing, endogenous NO would gradually transfer from peripheral vascular metastasis to the central vascular and with the growth of cloves of garlic scape, not much change in distribution and the level of NO. Exogenous NO may induce endogenous NO level increased, which maybe through the ways of nitric oxide synthase.
5. H_2O_2 Participate in postharvest nutrient re-distribution of garlic scape. Endogenous H_2O_2 has distributed in entire tissue of garlic scape throughout the transfer process, but in the aging tissues the congruent of H_2O_2 was no or very low. The H_2O_2 was very hige in eugenic apical tissue; High concentrations of H_2O_2 promoted redistribution of cell content and reduced the content of H_2O_2 in the basal garlic scape tissues could inhibit this action.
6. Inhibition effect of endogenous NO on postharvest nutrient re-distribution of garlic scape maybe regulate to the endogenous H_2O_2 level in garlic scape.
本实验利用不同浓度的一氧化氮(nitric oxide,NO)供体硝普钠(sodium nitro prusside,SNP)及NO和H_2O_2清除剂抑制剂处理,采朋生理生化方法结合激光共聚焦扫描显微镜技术,初步探索NO对蒜苔采后物质再转运作用机制。
实验结果表明:
1.适宜浓度SNP处理对蒜苔的保鲜效果显著。能够延缓蒜苔苔苞膨大、基部可溶性糖、可溶性蛋白含量的下降以及纤维化程度的升高,且苔茎保持鲜绿。其中,0.1mmol·L~(-1)的SNP浸泡苔茎基部3h效果最佳。高于0.5 mmol·L~(-1)的SNP处理,对蒜苔造成伤害。
2.SNP处理能显著地抑制苔苞直径的膨大及H_2O_2含量的增加。在4±1℃冷藏过程中,0.1mmol·L~(-1)的SNP处理抑制了苔苞膨大、苔茎基部H_2O_2含量增加、相对膜透性上升和POD活性的提高;提高了SOD、CAT和抗坏血酸过氧化物酶(APX)和顶部POD活性。
3.APase酶活性在贮藏前期有很高的活性,但随着贮藏时间的推移,活性呈下降趋势;SNP处理能够延缓APase酶活性高峰期的出现。
4.蒜苔中内源NO分布和内含物转运的程度有关:转运程度比较低的组织中分布比较均匀,随着转运程度的升高,逐步会由周缘维管束向中央维管束转移,但是,顶端组织NO分布和水平没有太大变化;外源NO处理能够诱导内源NO水平升高,这可能是通过NOS途径起作用。
5.H_2O_2参与蒜苔的物质转运过程。在整个转运过程中,H_2O_2在所有组织中都有分布,衰老组织中H_2O_2含量很少或者没有。生长旺盛顶端组织中,水平很高;高浓度的H_2O_2促进物质转运,降低基部H_2O_2含量,可以抑制物质转运。并且在物质转运过程中二者相互影响。
6.在蒜苔物质再转运过程中NO抑制物质转运的作用与内源H_2O_2的水平有关。
NO as a new type of intercellular and intracellar signaling molecule messaging become a research hotspot. NO has been found the role of delaying in a variety of fruits and vegetables senescence. Redistribution of cell in excised garlic scape lead to decline in eating quality, of such models of the aging mechanism of fruits and vegetables is unknown. In this study, garlic scape as the typical models of transport of substances, study the effects of NO on the role of senescence mechanisms transduction in excised garlic scape, in order to provide scientific basis to solve this kind of horticultural products of storing and keeping fresh problems.
Using sodium nitro prusside (SNP) as a nitric oxide (NO) donor, Hemoglobin (HB) as a NO scavenger, N~ω-nitro-L-arsinine methylter (L-NAME) as a NOS inhibitor, Sodium Tungstate as a NR inhibitor, Diphenyl iodine (DPI) as a NADPH oxidase inhibitor, Ascorbic acid (ASA) as a H_2O_2 scavenger, Physiological and biochemical methods combined with confocal laser scanning microscopy technology, Preliminary exploration of mechanisms of NO on transport of substances in excised garlic scape. The results as follows:
1. The results showed that suitable concentration of SNP had a remarkable effect on garlic scape preservation. It can inhibited the diameter of garlic bulbet and the degree of fibrosis increased, delayed the content of soluble suger and soluble protein in the basal of garlic scape, and the garlic scape were keeped viridity. 0.1 mmol·L~(-1) SNP solutions three hours at the basal of garlic scape was the best effect concentration in Preservation. But overhigh concentration of SNP (0.5mmol·L~(-1)) will cause the garlic sacape tissues injuries.
2. SNP treatment could significantly inhibit the increased of the diameter of garlic bulbet and the content of H_2O_2. 0.1 mmol·L~(-1) SNP inhibited the increased of the diameter of garlic bulbet and the content of H_2O_2 in basal garlic scape, relative membrane permeability increased, activities of peroxidase (POD) in basal garlic scape increased and promoted the activity of catalysis of superoxide dismutase (SOD), catalysis of catslase (CAT), ascorbate peroxidase (APX) in garlic scape and POD in apical garlic scape during four centigrade degree cold storage.
3. The acid phosphatase activities in the clove and the basal scape were very high, especially in the earier period of the pre-storage. However, with the time of torage, the acid phosphatase activities were a downward trend; Exogenous NO obviously slowed down the peak of acid phosphatase activities duing storage.
4. Distribution of endogenous NO related to the speed and extent of redistribution of cell content. Endogenous NO distributed evenly in low degree of transport of tissues, with the degree of transport increasing, endogenous NO would gradually transfer from peripheral vascular metastasis to the central vascular and with the growth of cloves of garlic scape, not much change in distribution and the level of NO. Exogenous NO may induce endogenous NO level increased, which maybe through the ways of nitric oxide synthase.
5. H_2O_2 Participate in postharvest nutrient re-distribution of garlic scape. Endogenous H_2O_2 has distributed in entire tissue of garlic scape throughout the transfer process, but in the aging tissues the congruent of H_2O_2 was no or very low. The H_2O_2 was very hige in eugenic apical tissue; High concentrations of H_2O_2 promoted redistribution of cell content and reduced the content of H_2O_2 in the basal garlic scape tissues could inhibit this action.
6. Inhibition effect of endogenous NO on postharvest nutrient re-distribution of garlic scape maybe regulate to the endogenous H_2O_2 level in garlic scape.
引文
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