Study on Antioxidant Enzyme Activity and Physiology and Biochemistry of Solanum nigrum L. under Glyphosate Stress

[Objectives] This study was conducted to investigate the scientific prevention and control of Solanum nigrum L. [Methods] Through experiments on S. nigrum from different sources, it was found that glyphosate stress had significant effects on antioxidant enzyme activity and oxidative damage of sensitive S. nigrum plants. [Results] Sensitive S. nigrum showed oxidative damage under glyphosate stress, while resistant S. nigrum responded to adversity damage by improving its antioxidant enzyme activity. The experimental results showed that the antioxidant enzymes and reduced glutathione of S. nigrum had certain metabolic detoxification effects under glyphosate stress. [Conclusions] This study provides a theoretical basis for scientific prevention and control of S. nigrum , and has a certain reference value for revealing the glyphosate resistance mechanism of S. nigrum .

镉对龙葵(Solanum nigrum L．)根系形态及部分生理指标的影响

以龙葵（Solanum nigrum L）为材料，水培法设置了5个镉质量浓度：0、10、25、50、100mg·L^-1进行试验。在处理17d和34d时，分别对其根系形态和部分生理指标进行测定。结果表明：（1）中低质量浓度（10～50mg·L^-1）的镉促进龙葵根系生长，表现在处理17d时其根长、体积均随镉质量浓度升高而增加；高质量浓度（100mg·L^-1）的镉抑制根系生长，表现在处理34d时其根长、体积、直径低于对照，根长达到届著水平。随着镉处理时间的延长，10mg·L^-1促进效果明品，表现为处理34d时其根长、体积、直径均达到最大值，100mg·L^-1抑制程度加剧，表现为其根长、体积、直径均急剧下降；（2）0～50mg·L^-1的Pro含量和质膜透性与对照相比差异不显著，100mg·L^-1 MDA与Pro含量均冠著高于对照，100mg·L^-1的质膜透性在处理34d时比对照增加了58．7％。说明高质量浓度镉使龙葵积累了大量的MDA和Pro，对龙葵表现出毒害作用，而龙葵作为超积累植物对镉胁迫表现出一定的耐性与适应性。

Stress Effect of Cadmium Absorption between Aloe and Solanum nigrum L

The stress effect of cadmium absorption between aloe and Solanum nigrum L was studied through a simulation experiment with different cadmium contents in soil. The results showed that the growth characteristics, biomass and Cd content in the plants of aloe and S. nigrum L were closely related to Cd content in the soil, and S. nigrum L and aloe had a strong tolerance to Cd in the experiment, pH of rhizosphere soil with S. ni- grum L and aloe planted was generally higher than that of non-rhizosphere soil, which is related to the planting pattern. Compared with aloe, S. nig- rum L had a stronger capacity to accumulate Cd, and intercropping S. nigrum L and aloe could inhibit aloe＇s absorption of Cd, which is favorable to the safe planting of aloe. In addition, S. nigrum L has the potential to restore polluted soil.

龙葵(Solanum nigrum L.)超富集镉的生理和分子机制研究进展

超富集植物在重金属污染土壤植物修复中具有重要的应用前景,已成为一种重要的生物资源。自2005年发现龙葵(Solanum nigrum L.)能超量富集镉以来,中国国内外围绕其超富集镉的机制和土壤修复应用开展了很多研究。本文从根系对镉的快速吸收、镉从根到地上部的有效转运以及较强的体内解毒功能(如植物细胞的区隔化、抗氧化、有机酸生成、渗透物质调节、光合和呼吸作用的维持以及氮代谢调节等)3个方面,详细综述了龙葵超富集镉的生理和分子机制。在总结相关研究成果的基础上,展望了该领域今后的发展趋势,建议应结合光合生化模型研究龙葵对镉的光合响应机制,并通过基因编辑技术提高修复效率和优化风险评估效果,同时加强镉胁迫下龙葵信号通路的精细调控等方面研究,以期为深入揭示龙葵超富集镉的机制提供参考。

Screening of Macroporous Resin for Polysaccharide of Solanum nigrum L.

[Objectives] To screen the optimal macroporous resin for polysaccharide of Solanum nigrum L. [Methods] The polysaccharide content was taken as the indicator, and adsorption and desorption performance of 20 kinds of resin were observed and studied. [Results] Adsorption and desorption performance of 20 kinds of resin had significant difference. S-8 type macroporous resin showed the best comprehensive property. [Conclusions] S-8 type macroporous resin should be used to purify polysaccharide of S. nigrum L.

Antioxygenic Activity of <i>Solanum nigrum L.</i>Leaves in Sunflower Oil Model System and Its Thermal Stability

The antioxygenic activity of Solanum nigrum L. leaves and its various solvent extracts were evaluated using sunflower oil model system. Leaf powder and its methanol/water (80:20) soluble fraction showed strong antioxygenic activity in refined sunflower oil. On the other hand, ethyl acetate fraction exhibited marginal antioxygenic activity, whereas the water soluble fraction was practically devoid of any activity in refined sunflower oil. Thermal stability of different extracts of Solanum nigrum L. leaves heated at 80?C in refined sunflower oil also indicated the strong efficacy of methanol/water (80:20) extract to inhibit thermal oxidation. Solanum nigrum L. contain high levels of magnesium (239.0 mg/100g) and phosphorous (80.3 mg/100g). Fatty acid analysis of the lipid extracted from Solanum nigrum L. leaves indicated the presence of linoleic (59.1%) as a major fatty acid. The result of this study confirmed the presence of antioxygenic compounds in leaves, in particular its methanol/water (80:20) extracts showed great potential as a natural antioxidant to inhibit lipid peroxidation in foods.

龙葵(Solanum nigrum L.)野外场地规模Cd污染土壤修复试验

镉（Cd）是我国重金属污染土壤中主要的污染元素之一。在污染土壤的修复措施中,植物修复是一种绿色技术,具有传统土壤治理方法所无法比拟的优点,也是现今国际环境修复的热点和前沿领域。本研究以龙葵为修复植物,进行了为期2年的野外场地规模Cd污染土壤植物修复预试验和试验研究。对修复前、后土壤Cd含量分析表明,土壤表层和亚表层Cd的去除作用明显。预试验和重复试验中,土壤表层Cd的平均减少率为6.3%和16.8%,亚表层各层Cd的减少幅度分别为50.6%和49.5%（20～40 cm）、73.5%和53.9%（40～60 cm）、80.7%或未检出（60～80 cm）。2组数据表明,在农田土壤条件下,龙葵植株可产生较大的生物量,从而提高对Cd的积累与运移能力;采用植物修复技术可实现轻、中度Cd污染土壤的修复,以实现农业生产的良性循环。

外源表油菜素内酯对镉胁迫下龙葵(Solanum nigrum L.)的缓解作用

为了解外源表油菜素内酯(EBL)对龙葵镉(Cd)胁迫的调节作用,对100μmol L-1 Cd胁迫处理7 d和20 d后的龙葵喷施10-9、10-5 mol L-1 EBL 7 d,研究其生长状况、不同器官的生理生化作用以及3个泛素基因(Rma1、BUB、UBC)的表达变化.结果表明:与对照相比,Cd胁迫下龙葵叶片和根部MDA含量、抗氧化酶(POD、SOD、CAT)活性升高,表明Cd对龙葵有一定的毒害作用.与单一Cd胁迫相比,外源EBL(10-9、10-5 mol L-1)处理下龙葵株高、根长、干重、鲜重显著(P<0.05)增加,而叶绿素含量增加不显著.与单一Cd胁迫7 d相比,EBL(10-9、10-5 mol L-1)+Cd处理组叶片和根部抗氧化酶活性增加不显著,其中10-9 mol L-1 EBL+Cd处理组叶片MDA含量显著(P<0.05)降低,泛素基因UBC、Rma1表达上调;10-5 mol L-1EBL+Cd处理组叶片Rma1表达上调.与单一Cd胁迫20 d相比,10-9 mol L-1 EBL+Cd处理组叶片和根部MDA含量降低显著(P<0.05),叶片和根部SOD、CAT活性以及叶片POD活性增加显著(P<0.05),叶片Rma1基因和根部BUB、UBC基因表达上调;10-5 mol L-1 EBL+Cd处理组叶片BUB、UBC基因和根部UBC基因表达上调.本研究表明Cd胁迫后,EBL通过提高叶片和根部抗氧化酶系统活性,减轻膜脂过氧化程度来缓解Cd胁迫对龙葵的损伤,并且泛素Rma1、BUB、UBC基因参与该过程,且表达存在时空特异性;当Cd胁迫20 d时,以10-9 mol L-1 EBL对龙葵的缓解作用更为显著.

The effects of different electrode materials on seed germination of Solanum nigrum L.and its Cd accumulation in soil

The effects of different electrode on Solanum nigrum L.seed germination were determined.The result showed that germination percentage(GP)of seeds in treatment T2(titanium elec-trode)was 26.6%higher than in control(CK,without electric field).High potassium and calcium concentrations were beneficial for seed enzymatic activity in treatment T2,which could partly explain the increase in GP.Cd accumulation(μg/pot)in S.nigrum treated with any electric field was significantly higher(p<0.05)than in CK without electric field.Specif-ically,Cd accumulation under the treatment T3(stainless steel electrode)was the highest both in roots and shoots;this accumulation in shoots and roots were 74.7%and 67.4%higher for stainless steel than in CK.This increase must have been associated with a higher Cd concentration in plants and did not exert a significant effect on the biomass.In par-ticular,Cd concentrations in roots and shoots under stainless steel treatment were both significantly higher than in CK(p<0.05),which had to be related to the higher available Cd concentration in the soil in the middle region.Furthermore,it could be attributed to al-tered soil pH and other soil properties.Moreover,none of the biomasses were significantly affected(p<0.05)by different electrode materials compared to CK.

2,4-EBL对不同浓度镉胁迫下龙葵(Solanum nigrum L.)幼苗的缓解作用

为探究外源2,4-表油菜素内酯(2,4-Epicastasterone,2,4-EBL/EBL)对镉(Cd)胁迫下龙葵幼苗的影响作用。本研究以Cd超积累植物龙葵为试验材料,采用水培法,对龙葵幼苗进行不同浓度的Cd胁迫处理(0,300μmol/L,600μmol/L,1 200μmol/L)后,对其叶片喷施浓度为10^-5mol/L的EBL溶液,研究外源EBL对Cd胁迫下龙葵幼苗生长的影响,以及过氧化物酶(peroxidase,POD)、过氧化氢酶(catalase,CAT)与超氧化物歧化酶(superoxide dismutase,SOD)的活性变化,丙二醛(malondialdehyde,MDA)和可溶性糖含量及叶绿素荧光动力学参数的变化。结果表明:(1)单一Cd胁迫下,龙葵幼苗株高、鲜重、干重及SOD、POD活性呈现低促高抑趋势,随着Cd浓度的增加,抑制根的伸长,增加了MDA含量、可溶性糖含量及CAT活性,降低了PSⅡ潜在荧光活性Fv/Fo和初始光化学效率Fv/Fm。(2)EBL降低了不同浓度Cd胁迫后MDA含量,提高了根部可溶性糖含量和抗氧化酶活性及叶片PSⅡ反应中心功能。研究发现外源EBL有利于龙葵幼苗在重金属Cd胁迫下提高其抗氧化酶活性,维持膜质氧化程度、光能的捕获与转换能力,促进龙葵幼苗的生长,有效缓解Cd胁迫对龙葵幼苗生长的抑制作用,且600μmol/L Cd胁迫后喷施EBL后期(12~24 h)对龙葵幼苗缓解效果最佳。本研究为外源施用EBL提高植物耐重金属Cd能力提供理论依据。

A newly-discovered Cd-hyperaccumulator Solanum nigrum L.

A systematic investigation was conducted to screen for cadmium-hyperaccumulator from 54 species in 20 weed families using outdoor pot-culture experiment and small-scale field experiment. The results from the outdoor pot-culture experiment showed that Cd concentrations in the stems and leaves of Solanum nigrum L. growing in a soil spiked with 25 mg/kg Cd were up to 103.8 and 124.6 mg/kg (DW), respectively, which was greater than 100 mg/kg, minimum Cd concentration for a Cd-hyperaccumulator. The Cdenrichment factor(EF,concentration ratio in plant to soil) in shoots was as high as 2.68. Moreover, Cd accumulation in shoots was greater than that in roots (TF, concentration ratio in shoots to roots) and the plant biomass growth was not inhibited at the Cd concentrations tested compared with the control. The results of the small-scale field experiment also showed that the characteristics of Cd accumulation in S. nigrum were all consistent with the characteristics of Cd-hyperaccumulators. Thus S. nigrum can be classified as a Cd-hyperaccumulator. This work is important for further research in the areas of hyperaccumulators screening, and plant-tolerance physiology and evolution. It provides a pat- entable new plant species for phytoremediation of Cd-con- taminated soils.

Effect of Polyethylene (LDPE) Microplastic on Remediation of Cadmium Contaminated Soil by <i>Solanum nigrum</i>L.

Solanum nigrum L. has a delightful prospect as a hyperaccumulation plant for cadmium pollution remediation, and microplastic is a new type of pollution that has received wide attention. In this study, the effects of polyethylene microplastics (LDPE) (0.135, 0.27, 0.81 and 1.35 mg·kg-1) and cadmium (20 mg·kg-1) on the growth indexes and soil physical and chemical properties of Solanum nigrum L. were investigated in a 17-day microcosm experiment. The results showed that single LDPE contamination showed a trend of low concentration promotion and medium-high concentration inhibited the growth index of Solanum nigrum L. and soil physicochemical index, and single Cd contamination was more stressful to plants than single LDPE contamination, while low concentration of LDPE could reduce the effect of Cd on soil physical and chemical properties and promote plant growth and uptake of soil nutrients. These findings suggest that high concentrations of microplastics can inhibit the growth of Solanum nigrum L. alone or in combination with Cd, rather than reducing the toxicity of Cd.

Cd污染河道底泥修复植物优选

选用12种植物对上海市某Cd污染黑臭河道底泥进行了修复试验研究,考察了修复植物对Cd污染底泥的耐受性、富集转运特性和修复效果,以期为实际开展植物修复Cd污染河道底泥研究提供科学依据.结果表明:除移植的灯芯草(Juncus effusus L.)生长情况较差外,其余11种植物对试验底泥均有良好的耐受性;龙葵(Solanum nigrum L.)、灯芯草、金盏菊(Calendula officinalis L.)、八宝景天(Hylotelephium erythrostictum(Miq.)H.Ohba)以及大吴风草(Farfugrium japonicum(L.f.)Kitam.)地上部分的Cd质量比均高于4.50 mg/kg;苘麻(Abutilon theophrasti Medicus)、灯芯草、金盏菊、八宝景天、大吴风草以及美洲商陆(Phytolacca americana L.)对Cd的富集系数(bioaccumulation factor,BCF)与转运系数(translocation factor,TF)均大于1;龙葵、金盏菊、美洲商陆以及三叶鬼针草(Bidens pilosa L.)对Cd的提取量均高于0.30 kg/hm^(2),具有较好的修复效果,其中龙葵对试验底泥中Cd的提取量为2.84 kg/hm^(2),提取效率可达88.75%,展现出了较大的修复潜力.优选试验结果证明,龙葵可作为该地区Cd污染河道底泥的最佳修复植物.

龙葵氯仿及正丁醇提取物对lewis肺癌移植瘤增殖及其血清IFN-γ、IL-2和IL-4含量的影响

目的:观察龙葵氯仿及正丁醇提取物对Lewis肺癌移植瘤生长及其外周血清IFN-γ、IL-2和IL-4的影响。方法:建立Lewis肺癌移植瘤模型,进行不同剂量龙葵氯仿及正丁醇提取物的体内抑瘤实验,观察抑瘤率;ELISA法测定荷瘤小鼠血清IFN-γ、IL-2和IL-4的含量。结果:经龙葵氯仿及正丁醇提取物用药的荷瘤小鼠,其平均瘤重均有不同程度的降低,与荷瘤空白对照组瘤重相比较差异均有统计学意义(P<0.01或P<0.05),其中氯仿中剂量组和正丁醇高剂量组的抑瘤率分别为38.93%和32.14%。不同剂量的龙葵氯仿及正丁醇提取物组可升高荷瘤小鼠血清IFN-γ、IL-2水平,降低IL-4水平,与荷瘤空白对照组比较差异均有统计学意义(P<0.01或P<0.05)。其中以龙葵正丁醇提取物高剂量和氯仿提取物中剂量组作用最为明显,与该两种提取物的抑瘤作用有一致性。结论:龙葵氯仿及正丁醇提取物能抑制Lewis肺癌移植瘤的生长,并可能使荷瘤小鼠Th1/Th2平衡向Th1偏移,在一定程度上恢复荷瘤机体抗肿瘤的免疫性反应。

龙葵提取物对荷lewis肺癌小鼠肿瘤组织细胞周期调控相关蛋白及凋亡的影响

[目的]观察龙葵氯仿及正丁醇提取物对荷lewis肺癌小鼠肿瘤组织细胞周期调控相关蛋白、细胞凋亡及相关凋亡蛋白表达的影响。[方法]建立lewis肺癌模型,64只C57BL/6J小鼠随机分成模型组、环磷酰胺组、龙葵氯仿和正丁醇提取物高、中、低剂量组,给药15d后处死,剥离肿瘤组织,称重,计算肿瘤生长抑制率;TUNEL染色法检测细胞凋亡;免疫组织化学法检测PCNA、p53、p21和Bax、Bcl-2。[结果]龙葵氯仿及正丁醇提取物对荷瘤小鼠肿瘤增殖有一定抑制作用,其中氯仿提取物中剂量组和正丁醇提取物高剂量组的抑瘤率分别为38.93%和32.14%（P〈0.01或P〈0.05）。龙葵氯仿及正丁醇提取物可降低荷瘤小鼠肿瘤组织PCNA和P53阳性细胞表达率,分别为29.7%-58%和32%-56%,并提高P21阳性细胞表达率（20%-32%）（P〈0.01或P〈0.05）。荷瘤小鼠肿瘤组织细胞的凋亡率均不同程度升高（15%-38%）（P〈0.01或P〈0.05）。Bax阳性细胞表达率均有不同程度升高（11%-41%）,Bcl-2阳性细胞表达率则均有不同程度降低（23%-37%）（P〈0.01或P〈0.05）,并可不同程度上调荷瘤小鼠肿瘤组织Bax/Bcl-2的比值。[结论]龙葵正丁醇及氯仿提取物有一定抑制lewis肺癌的生长作用,其作用机制可能与对荷瘤小鼠肿瘤组织细胞周期调控相关蛋白（PCNA、p53、p21）表达的影响、上调肿瘤组织Bax蛋白的表达、下调Bcl-2蛋白的表达,上调Bax/Bcl-2的比值;促进肿瘤组织的细胞凋亡等相关。

龙葵果HPLC指纹图谱研究

目的:建立龙葵果的HPLC指纹图谱分析方法,为该药材的鉴别及质量评价提供依据。方法:采用Agilent ZORBAX SB-C18色谱柱(150 mm×4.6 mm,5μm),以乙腈-0.3%磷酸溶液为流动相进行梯度洗脱,柱温为25℃,流速为1.0 m L/min,检测波长为205 nm,进样量10μL,分析龙葵果的HPLC图谱;采用"中药色谱指纹图谱相似度评价系统"(2004 A版)进行相似度评价。结果:建立了10批龙葵果药材的指纹图谱,确定了12个共有峰,指认了澳洲茄碱、澳洲茄边碱2个特征峰。结论:建立的龙葵果HPLC指纹图谱可为该药材的鉴别及质量评价提供更全面的参考。

澳洲茄碱抗肿瘤作用及机制研究进展

综述澳洲茄碱(solasonine)抗肿瘤作用及其机制的研究进展。澳洲茄碱对于肝癌、胃癌、肺癌、白血病、胰腺癌和大肠癌等疾病具有抗肿瘤作用,可以抑制癌细胞增殖、诱导癌细胞凋亡、抑制癌细胞迁移侵袭、诱导铁死亡、抑制糖酵解和降低癌细胞耐药性,可以调控Wnt/β-连环蛋白(Wnt/β-Catenin)通路、Hh信号通路、核因子-κB(NF-κB)通路、丝裂原活化蛋白激酶(AMPK)通路、磷脂酰肌醇3激酶/蛋白激酶B(PI3K/AKT)通路等的信号转导。

皖产药材龙葵的薄层色谱鉴别研究

目的采用薄层色谱法建立以薯蓣皂苷元为指标性成分的定性研究方法。方法依据中国药典2020年版四部通则(0502)的试验方法对其实验条件(以不同展开剂,不同提取方法,不同显色剂,不同点样量,不同薄层板)进行考察,最终确定其试验方法为取本品粉末2 g(过二号筛),加50 ml甲醇,超声30 min(频率45 kHz),滤过,滤液蒸干,残渣加10 ml的2 mol/L盐酸溶液,加热水解1 h,放冷,加三氯甲烷30 ml,加热回流30 min,萃取三氯甲烷液,水浴浓缩至2 ml。结果以硅胶G板为薄层板,以三氯甲烷-甲醇(20:0.2)为展开剂,以5%磷钼酸乙醇溶液为显色剂,在105℃加热至斑点显色清晰,在色谱图中,供试品和对照品的相应位置上显相同颜色的斑点。结论薄层色谱法的色谱斑点清晰,分离度好,可用于龙葵药材的薄层定性鉴别。

基于染色法和HPLC法测定龙葵不同部位中多酚含量

目的采用染色法测定龙葵不同部位的总酚酸含量;采用高效液相色谱(HPLC)法同时测定龙葵不同部位5个酚酸类化合物[绿原酸、咖啡酸、没食子酸、(E)-阿魏酸和原儿茶酸)]及1个黄酮化合物(芦丁)的含量。方法采用三氯化铁-铁氰化钾染色法测定龙葵不同部位的总酚酸含量;采用HPLC梯度洗脱,分段波长的方法同时测定6个多酚类成分的含量。结果染色法方法学结果显示总酚酸(没食子酸)在其线性范围内与测定值线性关系良好(r=0.9985),加样回收率为103.70%,RSD为2.7%,不同部位总酚酸以熟果中最高[(7040±195)μg·g^(-1),0.70%],其次为叶[(4430±120)μg·g^(-1),0.44%]、青果[(2720±97)μg·g^(-1),0.27%],茎中最低[(810±9)μg·g^(-1),0.08%]。HPLC方法学结果显示各成分在各自线性范围内与峰面积线性关系良好(r≥0.9993),加样回收率在95.58%~108.41%,RSD在0.34%~4.5%。结论染色法和HPLC法准确、稳定、可靠,可为龙葵药材的质量控制及物质基础研究提供依据。

HPLC同时测定新疆少花龙葵中绿原酸和芦丁的含量

目的：采用超声波提取,建立同时测定新疆少花龙葵中绿原酸和芦丁含量的高效液相色谱法.方法：色谱柱为安捷伦HC-C18柱（250 mm×4.6 mm,5μm）;流动相为甲醇-0.4％磷酸（50：50）;检测波长为360 nm;柱温为30℃;流速为1.00 mL·min-1,低压梯度洗脱.结果：绿原酸和芦丁质量浓度分别在6.25～ 150 μg·mL-1（r=0.999 8）和6.25 ～ 150 μg·mL-1（r =0.999 9）呈良好的线性关系,平均加样回收率分别为100.2％ （RSD=1.3％,n=5）和100.9％（RSD=1.4％,n=5）.结论：该方法简便快速、重复性好、准确、可靠,适用于同时测定新少花龙葵中绿原酸和芦丁含量.