摘要
甜菜红素在繁殖器官如花瓣、雄蕊、种子和果实特定的发育阶段合成;也存在于营养器官中,但受环境因子的高度调控。有关甜菜红素的早期研究,主要集中在影响其合成的相关因子方面,主要包括生长调节物质、光照、光质(远红光、红光、蓝光)、温度及营养元素(K+、NO3+、PO43-、蔗糖及Cu2+)等。
盐地碱蓬(Suaeda salsa L.)是典型的稀盐盐生植物,生长在黄河三角洲滨海潮间带和洼地生长的盐地碱蓬由于受低氧、高盐与低温等综合逆境胁迫,在整个生长季节地上部分皆为紫红色,而在地势较高或距海边较远的重盐碱地生长季地上部分皆为绿色。王长泉等已鉴定潮间带生长的盐地碱蓬积累的色素为甜菜红素。我们把两种表型盐地碱蓬种子都种在温室河砂中,其表型均为绿色,说明甜菜红素的积累主要是受环境因子诱导。作为一种真盐生植物,盐地碱蓬的耐盐机理已得到了比较广泛的研究,但有关盐地碱蓬积累甜菜红素的生物学意义尚未见报道。本论文目的是研究甜菜红素在抗氧化方面的功能,为盐地碱蓬的抗盐机理提供新的依据。
对盐地碱蓬进行低氧、不同盐及低温处理,以研究不同环境因子对盐地碱蓬甜菜红素含量的影响。结果表明:低氧处理2 h/d,处理60 d时,盐地碱蓬茎与叶甜菜红素含量最高;一定浓度的盐处理对盐地碱蓬甜菜红含量有促进作用,其中15 mmol/L MgSO4、20 mmol/L KCl和CaCl2及150 mmol/L MgCl2和KNO3作用显著。但有趣的是NaCl处理对盐地碱蓬甜菜红素的积累有抑制作用;低温(4±2℃)处理4 h/d,处理60 d时,甜菜红素含量最高;相同处理对盐地碱蓬体内甜菜红素合成的影响具有器官差异性,茎内甜菜红素含量高于叶片。这些结果表明:盐地碱蓬甜菜红素的合成受低氧、盐及低温等因子的综合影响,其中低氧和低温是主要因子。
在综合处理(15 mmol/L MgSO4、150 mmol/L MgCl2、20 mmol/L KCl、150 mmol/L KNO3、20 mmol/L CaCl2、缺氧处理2 h/d和低温处理4 h/d)条件下,茎内H2O2含量显著高于叶,说明过氧化氢可能是作为一个信号分子起作用的。茎与叶的DAB染色表明,H2O2对甜菜红素的诱导不仅与H2O2的含量有关,可能还与H2O2的维管运输或分布有关。在茎的形态学上部,甜菜红素含量低,而H2O2的含量却高,说明甜菜红素的合成过程或分子结构本身可能对H2O2存在清除作用。
无论是添加酪氨酸酶抑制剂处理,还是综合处理条件下茎与叶的比较,甜菜红素的含量皆有显著性差异,但甜菜红素的存在对抗氧化酶活性的影响却不尽相同。其中,APX酶活性增加,但SOD的酶活性升高或降低,而CAT酶活性则不变,说明甜菜红素可能对超氧阴离子亦存在清除作用。
以自然条件潮间带生境下的紫红色表型和绿色表型的盐地碱蓬为实验材料,对二者在抗氧化酶、过氧化氢含量等方面进行了比较研究,并对紫红色表型盐地碱蓬进行外源过氧化氢处理,分析了外源H2O2浓度与叶片甜菜红素含量的相关性。结果表明:绿色叶表型的盐地碱蓬叶内过氧化氢含量极显著高于紫红色叶表型盐地碱蓬,而二者的抗氧化酶(SOD、CAT和APX)的酶活性无显著性差异。外源过氧化氢的浓度与叶片内甜菜红素含量存在显著负相关。
以上结果表明,甜菜红素可能作为一种非酶促抗氧化剂参与过氧化氢水平的调节,从而减少胁迫引起的氧化伤害。
Betacyanin is synthesized in vegetative and reproductive organs of some plants. In vegetative organs, its accumulation is highly affected by enviromental condition, but in reproductive organs such as petals, stamens, seeds and fruits, betacyanin is mainly controlled by developmental stages. Previous studies were mainly focused on the factors, which influenced the accumulation of betacyanin, e.g. plant growth regulators, light quality (e.g. red light, far-red light and blue light), light quantity, temperature and nutrition elements (e.g. K+、NO3+、PO43-、sucrose and Cu2+).
The Chenopodiaceae Suaeda salsa L., a C3 plant, is a succulent euhalophyte. Shoots of S. salsa are violet-red throughout the whole growth period under hypoxia, high salt and low temperature conditions in the intertidal zone, but green on saline inland far from the seaside. Nowadays, the pigment in plants has been identified as betacyanin in S. salsa which occurs in the intertidal zone. In our previous investigations, when seeds of S. salsa from either violet-red phenotype plants grown in the intertidal zone or green phenotype plants grown in saline inland were sown in sand and cultivated in a greenhouse, their phenotype were green regardless of their mother plants. The result demonstrates that the accumulation of betacyanin is mostly regulated by enviromental factors. As a halophyte, physiological and molecular responses of Suaeda salsa to salinity stress have been extensively studied. But nothing is known about the antioxidant function of betacyanin accumulating in S. salsa.
In the present study, firstly, the effects of hypoxia, salt and low temperature on the betacyanin accumulation of S. salsa were investigated. The results indicated that the betacyanin contents were the highest in both stems and leaves of S. salsa plants treated with flooding for 2 h daily for 60 d. Different salts in a certain range of concentrations led to different betacyanin accumulation, e.g. 15 mmol/L MgSO4, 20 mmol/L KCl, 20 mmol/L CaCl2, 150 mmol/L MgCl2 and 150 mmol/L KNO3 gave rise to the higher betacyanin content, compared with control. Surprisingly, NaCl had an inhibitory effect on the betacyanin accumulation. Under the 4 h/d of low temperature (4±2℃) treatment for 60 d, the betacyanin of S. salsa was the hightest among the all treatments. Nevertheless, the effects of various treatments on the betacyanin content were different in different organs of S. salsa, e.g. the betacyanin content in stems was significantly higher than that in leaves. These results suggested that the accumulation of betacyanin in S. salsa is comprehensively influenced by different factors such as hypoxia, salts and low temperature, and hypoxia and low temperature is the main factor.
Under comprehensive consitions (15 mmol/L MgSO4, 150 mmol/L MgCl2, 20 mmol/L KCl, 150 mmol/L KNO3, 20 mmol/L CaCl2, hypoxia treatment for 2 h/d and low temperature treatment for 4 h/d), the H2O2 content of stem was significantly higher than that of leaf. The DAB staining of stem and leaf showed that the H2O2 induction for betacyanin was not only related to the content of H2O2, but also related to the H2O2 transport through vascular or its distribution. The result indicated that H2O2 may be acted as a signal molecule. At the morphological top of stem, the betacyanin content was lower, but the H2O2 content was higher. The biosynthesis of betacyanin or its molecular structure may have an activity of eliminating H2O2. Both under the treatment of the tyrosinase inhibitor, arbutin, and the comparision of stem and leaf treated with comprehensive condition, the betacyanin contents were significantly different from the control. But the existence of betacyanin had different effects on the antioxidant enzymes activities. The APX activity was increased, the SOD activity was increased or decreased, and the CAT activity was not affected. These results suggested that the betacyanin may have an activity of eliminating O2˙. In the present study, the leaves of red-violet phenotype and green phenotype S.
salsa grown in the same intertidal zone were used to comparatively study the activity of antioxidant enzymes (SOD, APX and CAT) and the content of H2O2, and to examine the correlation between the exogenous H2O2 concentration and betacyanin content of red-violet phenotype using detached leaves. The results showed that the H2O2 content of green phenotype S.salsa leaves was significantly higher than that of red-violet phenotype leaves, while no significant differences in the activity of antioxidant enzymes (SOD, APX and CAT) were detected between the two phenotypes of S. salsa. Correlative analysis showed that the exogenous H2O2 concentration was significantly negative correlated to betacyanin content of the detached leaves of violet-red phenotype S. salsa.
These results suggest that betacyanin possibly plays an important role as a non-enzymatic antioxidant in regulating the level of H2O2, sequentially alleviating the oxidative damage caused by oxidative stress.
引文
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