植物硒吸收转化机制及生理作用研究进展

硒是大多微生物、动物及人类的必要微量元素,但其在植物生长发育中的生理作用至今存在争议.较低浓度硒具有促进植物生长、提高植物耐受能力的功能,而大部分植物在高浓度下表现出中毒现象.随着人类对摄入硒及环境硒污染问题的认识加深,作物硒生物强化与硒污染植物修复问题引起重视,推动了对硒在植物中的吸收积累及代谢调控的研究.近年来对植物硒吸收及转化的研究表明,不同硒水平下植物对硒吸收积累及生理响应存在差异,土壤环境因素对植物硒吸收及转化具有重要影响,对高聚硒植物硒代谢研究逐渐揭示出硒在植物体内的转化过程和调控机理等.本文总结了目前硒生物强化与植物修复方面的研究进展,对环境中硒分布特点、植物硒吸收及其影响因素、植物体内硒转化及其过程调控关键酶,以及硒在植物中的生理作用等进行了综述,并对植物硒生理及分子机制未来研究方向进行展望.     

Study on the Relationship between Soil Selenium and Plant Selenium Uptake

Various extraction methods have been used to determine selenium (Se) concentrations in soils and plants in the second seleniferous regions of China. Our results show tea Se contents in the study area range from 1.009 to 2.6 mg/kg, which reveal that the tea areas in Ziyang County are in seleniferous regions. The four extraction methods evaluated in this study provide different information concerning soil and plant Se levels. The quality control/quality assurance program for this project indicated there is excellent agreement between total soil Se and extractable Se. For example, phosphate extractable Se results from the field investigation and greenhouse study were found to be highly correlated (R ² > 0.91) by linear regression analyses. Results from rye seedling experiments further show phosphate extractable Se has significant correlations with plant Se uptake and that a 0.1 M solution of KH₂PO₄ can be used as the extractant of soil available Se. In the acid soil, the Brassica campestris yield could be significantly reduced when the content of Se⁶⁺–Se ≥ 0.5 mg/kg, and the influence on the yield was not as obvious when the content of Se⁶⁺–Se reached up to 2.0 mg/kg. The uptake by Brassica campestris of Se⁶⁺–Se is higher than that of Se⁴⁺–Se. The main factors influencing the biological availability of soil Se, in order of their importance are CaCO₃, the presence of silt grains, organic matter and the presence of clay grains. pH could affect KH₂PO₄ extractable Se through CaCO₃.     

用蚯蚓溶酶体作为检测土壤污染的生物标志物

蚯蚓体腔细胞内的溶酶体能积累中性红染料.当受到环境胁迫时,溶酶体膜容易破裂,染料就逐步泄露到细胞质中,使细胞染成红色.中性红的保持时间(neutral red retention time,NRRT)可反映溶酶体受损情况.用不同浓度乙草胺和不同浓度Se处理赤子爱胜蚓(Eisenia fetida),观察体腔细胞溶酶体中性红染色的保持时间.结果表明,随乙草胺和Se处理浓度的增高和处理时间的延长,NRRT缩短;NRRT与土壤中乙草胺残留的浓度呈明显的负相关,与蚯蚓体内富集的Se也有明显的负相关.可见,蚯蚓溶酶体中性红染色保持时间是测定土壤污染物的生物标志物,在环境风险评价和土壤监测中具有稳定性和准确性,可作为早期预警生物标志物.     

苹果对硒的吸收及其积聚特征研究

对秦冠苹果采用不同浓度的硒溶液进行了花期喷施、对干注射和土壤施加３种处理,研究了苹果树对硒的吸收,找到了生产富硒苹果的有效方法。研究表明,硒并不是苹果必需的微显元素;首次提出了苹果树属非积聚硒植物的新概念;同时也对食用富硒苹果的有效性与安全性进行了分析     

中国硒资源利用中若干关键科学问题的探讨

中国是典型缺硒大国,一条马鞍形土壤缺硒带呈东北-西南走向分布,占主要农耕区土壤面积的33.34%,但同时也存在点状分布的富硒-高硒地区,占主要农耕区土壤面积的8.69%,成为湖北恩施、陕西安康、安徽石台、广西巴马、江西宜春等地方农业转型升级的新抓手,得到大力开发利用。伴随着硒资源的开发利用,一些科学问题被广泛提出,如：硒摄入有何健康效果？天然富硒区人群是否有实证研究数据？硒-镉共生导致天然富硒农产品富硒的同时是否存在镉含量超标问题？硒资源中的硒形态组成有何重要意义？硒超积累植物-壶瓶碎米荠的超积累硒的机制是什么？这些关键科学问题亟待解答。基于对天然硒资源近10年的研究成果,对以上科学问题进行了一些有益探讨,以期为未来高效安全科学地利用硒资源提供一些研究思路。     

Selenium uptake,speciation and stressed response of Nicotiana tabacum L.

The research on the function and mechanism of selenium (Se) is of great significance for the development of Se-enriched agricultural products. In this paper, uptake, speciation distribution, the effects on the flue-cured tobacco growth and antioxidant system of Se at different levels (0–22.2 mg Se kg⁻¹) were studied through a pot experiment, aiming to clarify flue-cured tobacco's response to Se stress and the relationship between Se speciation and antioxidant system. The results showed that the leaf area and number, the biomass and the chlorophyll content reached the maximum at 4.4 mg kg⁻¹ of Se treatment. Selenium at low levels (≤4.4 mg kg⁻¹) stimulated the growth of flue-cured tobacco by elevating the capability of antioxidant stress and reducing the malondialdehyde (MDA) content to 0.6–0.8 times of that of the control. However, high Se levels (≥11.1 mg kg⁻¹) depressed the capability of antioxidant stress and raised the MDA content to 1.5-fold of that of the control, and meanwhile the biomass of the aboveground parts and underground parts declined notably. The Se content in different parts of flue-cured tobacco significantly increased with the growth of Se levels. The range of Se content in roots, leaves and stems at 2.2–22.2 mg kg⁻¹ of Se treatment were 16.7–58.6 mg kg⁻¹, 2.6–37.3 mg kg⁻¹ and 2.2–10.3 mg kg⁻¹, respectively. According to the detection of different Se speciation, only selenocysteine (SeCys) was detectable in leaves at 2.2 mg kg⁻¹ Se treatment; SeCys, selenite [Se(IV)]and selenate [Se(VI)] were detected in flue-cured tobacco leaves at Se treatment (≥4.4 mg kg⁻¹), which accounted for 4.6–10%, 9–18.7% and 71–86% respectively; SeCys, selenomethionine (SeMet) and Se(IV) were detected in roots, and organic selenium(66–84%) was the main Se species at Se ≤ 11.1 mg kg⁻¹ treatment; four Se species [SeCys, SeMet, Se(IV) and Se(VI)] were detected in flue-cured tobacco roots, and the main Se species was inorganic Se (60%) at 22.2 mg kg⁻¹ Se treatment. That was to say, the percentage of organic Se species (SeCys and SeMet in flue-cured tobacco leaves and root) declined, whereas the ratio of inorganic Se species [Se(IV) and Se(VI)] increased with the growth of Se levels. The correlation analysis showed that the superoxide dismutase (SOD) activity as well as the glutathione (GSH) and MDA contents were positively correlated with the Se(IV) and Se(VI) contents at P < 0.01 and excessive inorganic Se might destruct the reactive oxygen species (ROS) balance and enhance the MDA content, thus causing damage to the plant growth. In a word, the present study suggested that the ratio of inorganic Se [Se(IV) and Se(VI)] was closely related with the growth and the antioxidant capacity of flue-cured tobacco and the excessive application of Se led to the higher proportion of inorganic Se and poorer antioxidant capacity, which ultimately inhibited the growth of flue-cured tobacco.     

硒缺乏与甲状腺激素代谢及功能

Ｉ型脱碘酶为含硒酶,缺硒时,该酶活性下降,使循环Ｔ４增高,外周组织Ｔ３水平下降。缺硒可加速甲状腺组织碘的耗竭,并加重某些缺碘的生物学效应,缺硒还可能与碘缺乏病的发生发展有关。以缺硒为主要病因的克山病存在甲状腺激素代谢异常,其心肌呼吸酶活性变化与缺碘引起的甲状腺功能低下相似,缺碘可缺硒引起的甲状腺激素代谢改变与克山病的发生可能有关。     

Effects of different water management on absorption and accumulation of selenium in rice

Rice is the staple food for more than half of the world's population, but selenium (Se) is low in many rice growing countries. Water management model affects rice soil pH and Eh, and then affects the bioavailability of Se in soil. A pot experiment was conducted to investigate the effects of water management on soil Se species, dynamics and selenium uptake by rice plants. Sodium selenite was added to the soil so that the soil selenium content reached 0.5 mg kg^?1 to study the effects of 3 different water management modes on soil selenium uptake by rice plants. These three modes are flood irrigation (F), aerobic irrigation (A) and alternate flood and aerobic irrigation (AFA). The results showed that flooded irrigation treatment increased the soil soluble selenium concentration, and the selenium in soil solution mainly existed in the form of selenite and selenomethionine selenium oxide. The content of selenium in grain was 2.44 and 1.84 times that of flooded irrigation treatment under A and AFA respectively. The content of selenium in straw was 1.32 and 1.58 times that of flooded treatment under A and AFA respectively. After rice grain enzyme hydrolysis, HPLC-ICP-MS analysis showed that Selenomethionine was the main selenium speciation in rice grains. This study showed that aerobic flooded treatment is one of the most effective ways to increase selenium content in rice field.     

Selenoenzyme activities in selenium- and iodine-deficient sheep

This study was conducted to evaluate the effects of single and combined deficiencies of selenium and iodine on selenoenzyme activities in sheep. Twenty-four male lambs were assigned to one of four semisynthetic diets: combined deficient A (Se⁻I), Se-deficient B (Se⁻I⁺), I-deficient C (Se⁺I⁻), and basal diet D (Se⁺I⁺). Thyroid hormones (T₃, T₄), thyroid stimulating hormone (TSH), and inorganic iodine (PII) were determined in plasma. Selenium and glutathione peroxidase activity (GSH-Px) were determined in erythrocytes, and tissue samples, including the thyroid, liver, kidney, and brain, were taken for selenoenzyme analysis. Plasma T₃, T₄, and TSH concentrations were similar in all groups. Type I deiodinase (ID-I) activity in liver and kidney remained unchanged in Se or I deficiency. In contrast, hepatic ID-I activity was increased by 70% in combined Se-I deficiency. Thyroidal cystolic GSH-Px (c-GSH-Px) and phospholipid GSH-Px (ph-GSH-Px) activities remained constant in both Se-deficient groups, whereas thyroidal c-GSH-Px activity increased (57%) in I deficiency. Type II deiodinase (ID-II) activity was not detectable in the cerebrum and cerebellum, whereas cerebellum Type III deiodinase (ID-III) activity was decreased in I deficiency and combined Se-I deficiencies. The results of the present study support a sensitive interaction between Se and I deficiencies in sheep thyroid and brain. Furthermore, the lack of thyroidal ID-I activity, the presservation of the thyroidal antioxidant enzymes, and the increases in hepatic ID-I indicate that a compensatory mechanism(s) works toward retaining plasma T₃ levels, mostly by de novo synthesis of T₃ and peripheral deiodination of T₄ in Se- and I-deficient sheep.     

两种螺旋藻在不同生长阶段的硒胁迫和生物有机化效应

在极大螺旋藻(S.maxima)和钝顶螺旋藻(S.platensis)不同生长阶段进行硒胁迫处理,分别从接种后第1d至第5d开始添加硒,并不断增加硒含量,至第7d使硒的累计添加量为1000mg·L^-1,形成5种不同硒胁迫(硒胁迫Ⅰ～Ⅴ),观察各种硒胁迫下螺旋藻的生物量及对无机硒的生物有机化的影响。结果表明:硒胁迫Ⅰ～Ⅳ对两种螺旋藻的生长影响不明显,而硒胁迫Ⅴ对螺旋藻生长有明显促进作用;藻体含硒总量和螺旋藻对无机硒的有机化率按硒胁迫Ⅰ～Ⅴ依次增加。首次提出硒胁迫强度概念,并用此较好地解释了有关实验结果。     

碎米荠及富硒产品中无机硒提取方法的建立

采用不同浓度的HCl、NaOH、KH_2PO_4、NaHCO_3、KCl、流动相及超纯水作浸提液提取富硒产品中的无机硒后,用高效液相色谱-原子荧光光谱(HPLC-AFS)联用仪测定Se(IV)和Se(VI),并对浸提时间、浸提温度、振荡频率、液料比等条件进行系统研究,建立了富硒植物碎米荠及富硒产品中无机硒的提取方法:无机硒的最佳提取剂为0.1mol/L KH_2PO_4溶液,提取方法为:称取样品0.10g于离心管中,加入0.1mol/L KH_2PO_4溶液至3mL,于70℃、1 500r/min条件下振荡提取30min,以3 000r/min转速离心10min,0.45μm滤膜过滤后上机测定,样品加标回收率为86.14%~117.60%,Se(IV)和Se(VI)均能定性定量检测,为富硒产品中无机硒测定方法标准的制定奠定了基础。     

类风湿关节炎患者自我效能水平及相关因素分析

目的：探讨类风湿关节炎患者自我效能水平及影响因素,为科学的护理干预提供依据。方法：采用一般自我效能感量表（GSES）对随机抽样的267例类风湿关节炎患者进行调查。测定调查患者的自我效能水平。结果：267例类风湿关节炎患者自我效能水平较低。结论：多种因素影响类风湿患者的自我效能水平,这些影响因素包括：患者文化程度,收入,医疗付费方式,病程等。     

Se和V_E与克山病

以克山病病区粮配成基础饲料,另在基础饲料中分别补充Ｓｅ或ＶＥ,或Ｓｅ＋ＶＥ喂养大鼠,在细胞及亚细胞水平上以Ｃａ代谢为主研究并比较了Ｓｅ和ＶＥ在克山病病因中的作用。测量了心肌细胞和心肌线粒体的Ｃａ代谢及有关指标、心肌线粒体能量转换功能及心肌组织自由基含量。结果表明,在低Ｓｅ病区粮中补充Ｓｅ或ＶＥ均能在一定程度上预防病区粮中致病因素对心肌细胞及线粒体的损伤;并且补充Ｓｅ或ＶＥ均能使心肌组织中自由基含量减少。提示Ｓｅ和ＶＥ是通过清除体内过量自由基预防细胞和线粒体的损伤的。但值得注意的是,实验中所用病区粮ＶＥ含量不低于甚至高于非病区对照粮,在低Ｓｅ情况下,所补ＶＥ的量需要相当大（如本实验中补充２００μｇ／ｇ）才能较明显地预防心肌细胞和心肌线粒体的损伤。通过对这些结果的分析,进一步肯定低Ｓｅ是克山病形成的重要因素之一。     

Effects of dietary tin and aluminum on selenium utilization by adult males

The main purpose of these studies was to determine whether the amounts of tin and aluminum that can enter foods during processing and storage are sufficient to affect the utilization of selenium by human subjects. Two 40-day balance studies were conducted. The eight adult males who participated in the first study lost significantly more selenium in their feces when fed a test diet containing 50 mg tin daily than when fed the control diet containing 0.1 mg tin daily. During the first study subjects tended to excrete less selenium in the urine when fed the test diet rather than the control diet. In the second study, the dietary treatments (5 and 125 mg aluminum daily) had no effect on the excretion and apparent retention of selenium by eight adult males.     

鄂西自治州黄芪等药用植物中硒等微量元素的测定

我们用原子吸收分光光度法和荧光分光光度法测定了鄂西自治州近20种药用植物的含硒量,并同时测定了其中锌、铁、锰、铜等人体必须的微量元素含量,结果显示,半数植物含硒量较高,所有植物中锌、铁、锰、铜含量丰富。     

铬、硒对水稻幼苗生长和生理的影响

以单一铬(Cr6+0～200μmol·L1)及硒(Se0～200μmol·L1)、铬硒混合液(铬100μmol·L1,硒50μmol·L1)处理水稻幼苗,研究不同处理和浓度对水稻幼苗生长和生理特性影响。结果表明(1)单一铬处理,随着铬浓度增加,植株生长明显受到抑制,铬毒害表现为株高、鲜重和干重受抑制,叶片黄色、叶绿素含量下降,体内SOD、CAT活性下降,POD活性上升,膜透性增大;(2)单一硒处理,50μmol·L1促进植株生长,100μmol·L1和200μmol·L1则抑制植株生长;(3)铬硒混合处理结果反映,硒有减轻水稻铬毒害的作用,表现为:减轻铬胁迫对株高、鲜重和干重增加的抑制,提高叶绿素含量,提高SOD、CAT活性,降低POD活性和膜透性。     

Effect of sodium selenosulfate on restoring activities of selenium-dependent enzymes and selenium retention compared with sodium selenite in vitro and in vivo

Sodium selenosulfate has been extensively used as a precursor of selenide ions in the preparation of nano Se-containing compounds. Its biological properties remain completely unknown. Sodium selenosulfate and sodium selenite were added to the medium of HepG2 cells and administered intraperitoneally at a dose of 0.1 mg Se/kg body weight to selenium-deficient mice, respectively. Both of the selenium compounds could increase the activities of glutathione peroxidase (GPx) and thioredoxin reductase (TrxR) in a dose-dependent manner in cells and efficiently restore selenium retention and activities of GPx and TrxR in mice. All of the variables were in correlation with the Se supply. There was no distinction in elevating activities of GPx and TrxR between selenosulfate and selenite in vitro. After a 2-d supply of selenosulfate, the activity of GPx in the liver was 65% (p < 0.001) and Se accumulations in the liver, kidney and blood were 64%, 86%, and 65%, respectively, of those treated with selenite (allp < 0.01). With the 7-d selenosulfate supplementation, the activity of GPx in the kidney and activities of TrxR in the liver and kidney were 88%, 75%, and 78%, respectively, of those treated with selenite (allp < 0.01); Se retentions in the liver and kidney were 85% and 93%, respectively of those supplemented with selenite (bothp < 0.01). These facts indicated that selenosulfate could be absorbed and utilized in the biological system. No difference in vitro demonstrated that selenosulfate could be absorbed and generate reduced selenide as efficiently as selenite. The differences between the two compounds in vivo were the result of other factors that affected selenosulfate utilization in tissues.     

Selenium content and distribution in rat tissues irradiated with gamma rays

The effects of supplementation with selenous yeast and ionizing radiation on selenium (Se) content and distribution were evaluated in rat tissues (liver, kidney, spleen, heart, muscle, blood, front brain, hind brain, hypothalamus, pituitary, adrenal glands, testes, and hair). This study had 16 Se-supplemented (0.5 μg Se/d) and 16 placebo adult male Wistar rats. One half of the animals (eight Se-supplemented and eight placebos) were irradiated with a single dose of 4.2 Gy from a Co-60 source and sacrificed 7 d after irradiation along with nonirradiated animals and analyzed for Se content determination. The data obtained showed that selenous yeast supplementation increased Se levels in rat tissues (highest increases in hypothalamus, 161%; hind brain, 126%; spleen, 110%; and adrenal gland, 105%). Ionizing radiation induced significant changes in Se content and distribution (decrease in liver, blood, hair, femoral muscle, spleen, and hypothalamus; increase in kidney, testes, adrenal glands, and brain of placebo group). Supplementation with selenous yeast reduces changes in Se content and distribution after irradiation. It seems that the animal tissue susceptibility to oxidative damage may be correlated to their ability to retain Se in tissues.     

野生二粒小麦籽粒含硒量的差异分析

采用氢化物-原子荧光法,对种植于黔中的来自以色列15个野生二粒小麦群体110个基因型籽粒含硒量进行检测分析.结果表明,15个群体含硒量差异极显著(P≤0.001);110个基因型籽粒硒含量和单粒硒含量分别为0.043～0.409 mg kg-1和0.008～0.125 mg seed-1,平均值分别为0.180 mg kg-1和0.046 mg seed-1.籽粒硒含量和单粒硒含量最低的基因型分别是Gamla群体的TZ120和Mt.Hermon群体的TZ8,最高的基因型分别为Bat-Shelomo群体的TZ36和TZ34.在群体水平上硒浓度差异明显,变异系数CV为9%～74%;斯皮尔曼秩相关分析表明,籽粒硒含量分别与该群体起源地海拔、年平均降雨量、平均干旱天数呈显著负相关,与年均温、8月均温、1月均温呈显著正相关.单粒硒含量的相关分析与籽粒硒含量的相似.野生二粒小麦籽粒含硒量的差异是长期适应环境的结果,其遗传多样性将为小麦硒营养机理研究和育种利用提供材料.