美洲商陆(Phytolacca americana L.)——一种新的Mn积累植物

通过野外调查和水培试验,研究了锰在美州商陆体内积累情况及对其生长发育的影响,发现并鉴定出美洲商陆(Phytolacca am ericana L.)是一种新的M n积累植物。调查结果发现,美洲商陆对土壤中高含量的锰有很强的忍耐、吸收和积累能力,叶片中M n含量为5160￣8000m g·kg-1,平均为6490m g·kg-1。营养液培养试验证明,美洲商陆对生长介质中的M n有很强的忍耐能力,当生长介质中M n浓度高达10m m ol·L-1,叶片中M n含量达到11.76g·kg-1,植株仍生长正常。地上部M n含量及其积累量均随生长介质中M n浓度的增加而增加,当生长介质中M n浓度为50m m ol·L-1时,叶片中M n含量达到47.06g·kg-1。结果表明,美洲商陆是我国发现的一种新的M n积累植物,为今后探明植物积累M n的机理和M n污染土壤的植物修复提供了一种新的种质资源。

不同生境美洲商陆(Phytolacca americana L.)根系差异表达蛋白筛选

某铀尾矿区生长着大量美洲商陆（Phytolacca americana L.）,目前还未见美洲商陆耐受及富集铀相关蛋白质组学研究报道。为了找到铀耐受及富集表达相关蛋白,建立了铀尾矿区和对照区2种不同生境美洲商陆根系蛋白质组研究技术。通过全蛋白质制备条件优化、2DE电泳条件摸索以及染色方法选择,得到了重现性好、分离效果和清晰度都很高的美洲商陆根系双向电泳差异凝胶图谱。经软件分析,筛选出了铀耐受及富集相关蛋白24个,上调表达11个、下调表达13个。

锰超富集植物垂序商陆(Phytolacca americana L．)的认定

针对目前国内锰超富集植物商陆属(Phytolacca)植物名称混乱的状态,在重新审查锰超富集植物——商陆(Phytolacca acinosa Roxb.)的野外标本和温室培养植物的基础上,比对中国科学院昆明植物所标本馆的腊叶标本,并查阅相关文献资料,以期正确认定锰超富集累植物的学名。结果表明,湘潭锰矿尾矿废弃地原生的锰超富集植物实为垂序商陆(Phytolacca americana L.),国内外相关研究论文中出现的商陆和美洲商陆实为垂序商陆的同物异名。这一植物名称的认定,将对避免锰超富集植物研究重复进行、保证相关研究正常有序开展具有重要意义。

超积累植物垂序商陆(Phytolacca americana L.)吸收锰机制的初步探讨

垂序商陆是中国境内发现的锰超积累植物,了解其对锰的吸收特性及机制可为植物修复提供基础信息.通过人工培养液温室培养方法,探讨了低锰浓度处理下垂序商陆吸收累积锰的特性;采用同步辐射荧光技术(SRXRF)分析了锰在垂序商陆根横截面的分布特征;应用能量代谢抑制剂2,4-二硝基苯酚(DNP)和矾酸钠(Na3VO4)、钙离子通道抑制剂氯化镧(LaCl3)研究了垂序商陆对锰吸收,并探讨了其吸收机制.结果表明,低锰浓度处理下,锰超积累植物垂序商陆仍然具有较高的吸收和累积锰的能力,5μmol·L-1低锰浓度处理下垂序商陆根、茎、叶锰含量(以干重计)仍可高达402、208、601 mg·kg-1.锰在垂序商陆根横截面中柱含量最高,其次为表皮,皮层锰含量最少,锰从皮层进入到中柱是一个从低浓度到高浓度的过程,表明垂序商陆对锰的吸收可能存在主动运输过程.DNP和Na3VO4对垂序商陆锰吸收有一定的抑制作用,表明锰进入细胞是需要消耗能量的,进一步说明垂序商陆对锰的吸收存在主动吸收的过程.钙离子通道抑制剂LaCl3抑制垂序商陆根对锰的吸收高达30%,表明垂序商陆对锰的吸收与钙离子通道密切相关.

垂序商陆配方颗粒的HPLC指纹图谱研究与商陆皂苷甲含量测定

目的为垂序商陆的质量控制提供参考。方法建立高效液相色谱(HPLC)法,测定19批垂序商陆配方颗粒指纹图谱并进行相似度评价、聚类分析(CA)和主成分分析(PCA),结合正交偏最小二乘-判别分析(OPLS-DA)筛查不同批次垂序商陆配方颗粒的差异成分;通过HPLC-ELSD测定配方颗粒中商陆皂苷甲成分。结果垂序商陆配方颗粒HPLC指纹图谱共标定了9个共有峰,通过与对照品比对,确定了峰1为腺苷、峰2为鸟苷、峰3为L-色氨酸、峰5为刺桐碱、峰8为商陆皂苷甲。19批垂序商陆配方颗粒指纹图谱相似度均在0.89以上,CA、PCA和OPLS-DA均将样品分为了3类,在OPLS-DA模型下,以VIP值大于1筛选出3个差异性标志物,分别为L-色氨酸(3号峰)、商陆皂苷甲(8号峰)和刺桐碱(5号峰)色谱峰。19批配方颗粒样品中商陆皂苷甲的含量在1.8~7.6 mg/g之间。结论所建立的HPLC指纹图谱,结合化学计量学分析方法和商陆皂苷甲含量测定方法,能较为全面地评价垂序商陆配方颗粒的质量。

美洲大蠊(Periplaneta americana L.)抗菌肽杀菌作用初步观察

报道了美洲大蠊(PeriplanetaamericanaL.)抗菌肽对大肠杆菌(EscherichiacoliK88)的作用过程.电镜观察表明,美洲大蠊抗菌肽首先使细菌的外层及细胞质膜损伤,形成开口,导致内容物外泄而死亡,最后菌体崩解成碎片.

Two New Steroidal Glycosides from Fermented Leaves of Agave americana

Two new spirostanol glycosides named agamenoside A and B, were isolated from the fermented leaves of Agave americana. Their structures were elucidated as (23S, 25R)-5a -spirostan-3b, 6a, 23-triol 3-O-a-L-rhamnopyranosyl-(13)- b-D-glucopyranosyl-(12)-[b-D- xylopyranosyl-(13)]-b-D-glucopyranosyl-(14)-b-D-galactopyranoside (1) and (25R)-5a-spiro- stan-3b, 6a-diol 3-O-b-D-glucopyranosyl-(12)-[ b-D-xylopyranosyl-(13)]- b-D-glucopyra- nosyl-(14)- b-D-galactopyranoside (2) by a combination of chemical and spectral methods.

Comparison between Decolorization Effects of Activated Carbon and Macroporous Resin on Periplaneta americana L. Skimmed Cream

[Objective] The aim of this study was to investigate the decolorization ef- fects of activated carbon and macroporous resin on Periplaneta americana L. skimmed cream and compare the advantages and disadvantages of the two decolorization technologies. [Method] Periplaneta americana L. skimmed cream was decolored with activated carbon and macroporous resin, and freeze-dried to collect solid decolorization products. By investigating the yield, decolorization rate, protein retention rate and decolorization operation process, the advantages and disadvantages of the two decolorization technologies were compared. [Result] Both activated carbon and macroporous resin can be used for decolorization. To be specific, macroporous resin-decolorization is superior in the yield and protein retention rate, while activated carbon-decolorization is superior in decolorization rate and decolorization operation process. [Conclusion] Macroporous resin-decolorization can be used if protein is the main ingredient required in the experiment, while activated carbon-decolorization can be used if protein is not the main ingredient required.

A New Pregnane Glycoside from Fermented Leaves of Agave americana

A new minor pregnane glycoside was isolated from the fermented leaves of Agave americana. Its structure was elucidated as (20S)-5a-pregnane-3? 20-diol 20-O--D-glucopyrano- side (1) by spectral methods.

Two New Spirostanol Steroidal Sapogenins from Fermented Leaves of Agave americana

Two new spirostanol sapogenins named agavegenin A and B were isolated from the fermented leaves of Agave americana L. Their structures were elucidated as (23S, 25R)-5a- spirostan-3b, 6a, 11a, 23-tetraol (1) and (23S, 25S)-5a-spirostan-3b, 23, 27-triol (2) by spectral methods.

HPLC-ELSD法测定商陆不同部位中商陆皂苷甲的含量

目的:建立 HPLC-ELSD 法测定商陆中商陆皂苷甲的含量。方法:采用 Diamonsil C_(18)(5 μm,250 mm×4.6 mm)色谱柱,甲醇-水(75:25)为流动相,流速1.0 mL·min^(-1);蒸发光散射检测,ELSD 漂移管温度:74 ℃,载气流速:2.0 L·min^(-1)。结果:商陆皂苷甲进样浓度在26.6～532.5分g·mL^(-1)范围内有良好的线性关系(r=0.9995);平均回收率(n=5)为98.5%,RSD 为1.5%。结论:本方法简便,结果可靠、准确,可用于商陆的质量控制。

拉巴豆替代控制对美洲商陆根际土壤微环境的影响

为探究拉巴豆替代控制对美洲商陆根际土壤微环境的影响,分别测定了美洲商陆单种及与拉巴豆按不同比例混种时根际土壤细菌、真菌和放线菌数量,及土壤全氮、全磷、全钾、碱解氮、速效磷、速效钾和有机质含量。结果表明:混种拉巴豆显著降低了美洲商陆根际土壤细菌数量、放线菌数量和所占比例、微生物总数、全磷、碱解氮、速效钾和有机质含量,显著提高了真菌所占比例,1∶3混种(S2L6)时细菌所占比例显著降低,1∶1(S4L4)和1∶3(S2L6)混种时真菌数量显著提高,全氮和速效磷含量显著降低。相关性分析表明根际土壤微生物数量和理化性质相互影响,由主成分分析综合排名可知1∶3混种时美洲商陆根际土壤肥力条件最差。综上所述,因为混种使美洲商陆根际土壤肥力降低,说明拉巴豆能够成功替代控制美洲商陆,1∶3混种最有利于美洲商陆的替代控制。

Antifeedant Activity of Three Plant Extracts Including P. acinosa against Pieris rapae

[Objective] The research aimed to study antifeedant activity of Phytolacca acinosa Roxb., Setaria viridis （L.） Beauv and Viola yedoensis Makino extracts against Pieris rapae. [Method] Activity material was extracted from S. viridis （L.）, P. acinosa and V. yedoensis using acetone cold soak method, and non-selective antifeedant activity of extracts to Pieris rapae larva was determined by using lobular plate addition method. [Result] The results showed that the acetone leaching agent of P. acinosa had most obvious antifeedant effects on Pieris rapae. The antifeedant rate were 74.53% and 82.34% at 24 and 48 h respectively. With the concentration increasing, the antifeedant effect of P. acinosa extracts increased. The antifeedant rate of 0.050 g/ml treatment was the highest, being 74.53% and 82.34% at 24 and 48 h. [Conclusion] P. acinosa could be studied and utilized as potential botanical insecticide.

美洲大蠊不同发育阶段蛋白质、重金属含量测定及脂溶性成分的GC-MS分析

为促进药材资源的综合开发利用,对美洲大蠊1龄、3龄、6龄以及成虫中蛋白质和4种重金属元素含量、脂溶性成分组成进行了测试。采用凯氏定氮法测定粗蛋白含量、ICP-MS测定重金属元素含量、GC-MS测定脂溶性成分及相对含量。美洲大蠊在不同生长发育阶段的粗蛋白含量为56.61%~67.38%;砷含量≤0.2714 mg/kg;汞含量≤0.1972 mg/kg;铅含量≤0.5482 mg/kg;镉含量≤0.1222 mg/kg;从脂溶性成分中鉴定30种化合物,其中5种为4个生长发育阶段共有成分。美洲大蠊成虫体内蛋白质达63%,是优质的饲料蛋白来源。根据《饲料卫生标准》相关规定,其虫体中汞含量超标,需要对美洲大蠊养殖过程所用的饲料配方进行优化;美洲大蠊体内以棕榈酸为代表的饱和脂肪酸是其表现脂毒性的重要因素,油脂中富含抗氧化酚类物质。

Ecological niche shifts affect the potential invasive risk of Phytolacca americana(Phytolaccaceae)in China

Background:Predicting the potential habitat of Phytolacca americana,a high-risk invasive species,can help provide a scientific basis for its quarantine and control strategies.Using the optimized MaxEnt model,we applied the latest climate data,CMIP6,to predict the distribution of potential risk zones and their change patterns for P.americana under current and future(SSP126,SSP245,SSP585)climate conditions,followed by invasion potential analysis.Results:The predictions of MaxEnt model based on R language optimization were highly accurate.A significantly high area of 0.8703 was observed for working characteristic curve(AUC value)of subject and the kappa value was 0.8074.Under the current climate conditions,the risk zones for P.americana were mainly distributed in Sichuan,Chongqing,Guizhou,Hunan,and Guangxi provinces.The contribution rate of each climatic factor of P.americana was calculated using the jackknife test.The four factors with the highest contribution rate included minimum temperature of coldest month(bio6,51.4%),the monthly mean diurnal temperature difference(bio2,27.9%),precipitation of the driest quarter(bio17,4.9%),and the warmest seasonal precipitation(bio12,4.3%).Conclusion:Under future climatic conditions,the change in the habitat pattern of P.americana generally showed a migration toward the Yangtze River Delta region and the southeastern coastal region of China.This migration exhibited an expansion trend,highlighting the strong future invasiveness of the species.Based on the predictions,targeted prevention and control strategies for areas with significant changes in P.americana were developed.Therefore,this study emphasizes the need of an integrated approach to effectively prevent the further spread of invasive plants.

美洲大蠊不同炮制品对CCl_4致小鼠急性肝损伤的保护作用

目的探索不同炮制方法下美洲大蠊对CCl_4致小鼠急性肝损伤的保护作用。方法采用CCl_4致小鼠急性肝损伤模型,各实验组按炮制给药方法分为空白组、模型组、原药组、清炒组、盐炙组、麸炒组、米炒组、联苯双酯组。连续给药1周后,除空白组外,各实验组腹腔注射0.2%CCl_4橄榄油溶液,禁食不禁水24 h后,摘除小鼠眼球取血,分离血清。测定血清谷草转氨酶(AST)、丙氨酸转氨酶(ALT)的活性;取肝脏测肝脏系数,固定于10%的甲醛溶液中,进行肝脏病理组织学考察。然后采用索式提取器测定各炮制组油脂含有量,采用考马斯亮蓝法测定各炮制组蛋白质含有量,采用苯酚-硫酸分光光度法测定各炮制组多糖含有量。结果与模型组比较,各炮制组与原药组均能显著降低小鼠肝脏系数,各炮制组能显著降低小鼠血清中AST、ALT水平(P<0.01),而原药组对小鼠血清中AST、ALT水平的降低与模型组比较不具显著性差异(P>0.05),HE染色表明麸炒组病变最轻。与原药组比较,各炮制组油脂含有量下降;蛋白质含有量以盐炙组最低,原药组最高;多糖含有量在盐炙组、清炒组均较高,在原药组、麸炒组、米炒组均较低。结论美洲大蠊各炮制品均能对CCl_4致小鼠急性肝损伤起保护作用,以米炒、麸炒组效果最佳,原药组效果最差。

HPLC法测定美洲大蠊药材中五种核苷类成分含量

目的用HPLC法测定美洲大蠊药材中五种核苷类成分含量。方法用石油醚对美洲大蠊粉末进行脱脂,以65%的乙醇为溶剂进行超声提取,乙腈溶解配制成美洲大蠊供试品溶液,使用Agilent色谱柱(9.4×250 mm;5μm),以纯水∶乙腈=95∶5为流动相,流速为0.5 mL/min,检测波长为254 nm,进样量取10μL,对照混合标品和单一标品对美洲大蠊供试品溶液进行含量测定。结果用HPLC法测定美洲大蠊药材中五种核苷类成分含量,灵敏度高,专属性好,可用于本品的含量测定。5种核苷浓度在一定范围内与峰面积呈良好线性关系(R^(2)>0.9941),精密度高,重复性好,稳定性佳,加样回收率均在97.58%~103.26%之间。结论该法实验过程简便、结果可靠、稳定,可用来测定美洲大蠊中核苷类成分的含量。

美洲大蠊提取物CⅡ-3 UPLC指纹图谱研究

目的:建立美洲大蠊提取物CⅡ-3超高效液相色谱(Ultra Performance Liquid Chromatography,UPLC)指纹图谱,评价不同批次CⅡ-3样品的质量。方法:采用Phenomenex Gemini 5u C18110A(250 mm×4.6 mm)色谱柱分析美洲大蠊提取物CⅡ-3;柱温28℃;检测波长214 nm;流动相为乙腈-水(0.1%甲酸),进行梯度洗脱。依据相似度评价结果和化学计量学方法进行不同批次CⅡ-3质量评价。结果:建立了12批CⅡ-3指纹图谱,标定共有峰21个,其中11批样品相似度大于0.900。结论:首次建立了美洲大蠊提取物CⅡ-3的UPLC指纹图谱,为CⅡ-3质量控制提供科学依据。

Cloning and Sequencing of the Pokeweed Antiviral Protein Gene and Its Expression in E. coli

The total RNA was isolated from pokeweed (Phytolacca americana) leaves using the method of guanidine isothiocyanite and used as a template to amplify the deleted mutant pokeweed antiviral protein (PAP) gene by RT-PCR and then the gene was cloned into the pGEMR-T vector. The sequencing results showed that the PAP gene consisted of 711nt, which was 99.6% identical to the PAP gene reported by Lin et al (1991). The IPTG-inducible expression vector containing the PAP gene was constructed and transferred into the E. coli strain BL21 (DE3)-plysS. A specific protein was produced after induction with 0.4m mol/L IPTG and its molecular weight was 26ku. The results of the double diffusion on the agar plate and the western blotting test showed that the protein produced in E. coli was highly identical with the PAP extracted by a Frenchman from French pokeweed leaves. These revealed that PAP gene was actually achieved and exactly expressed in E. coli.

美洲大蠊粪便提取物对大鼠离体心脏功能的影响研究

[目的]探究美洲大蠊粪便提取物(PAF)对大鼠离体心脏功能的影响。[方法]采用Langendorff灌流装置构建SD大鼠离体心脏灌流模型,检测各组左室收缩压(LVSP)、左心室压力最大上升和下降速率(±dp/dtmax)、心率(HR)等血流动力学指标的变化。[结果] PAF可显著提升正常大鼠离体心脏的LVSP和±dp/dtmax,且呈剂量依赖性,其中0.5 mg/mL的高剂量作用效果最为显著,但PAF对HR并未表现出明显的影响。[结论] PAF升高LVSP,增大±dp/dtmax,对大鼠离体心脏有正性肌力作用,能够增强心肌收缩力。PAF对心率未表现出明显影响,有利于降低心脏耗氧量,可能是其能够发挥心脏保护作用的重要原因。