黄花蒿(Artemisia annua L.)提取物对两种病原真菌的生物活性

以石油醚Ⅰ(30℃～60℃)、石油醚Ⅱ(60℃～90℃)、乙醇和丙酮等4种溶剂对黄花蒿的根、茎、叶进行初步提取,采用生长速率法测定不同提取物对玉米小斑病菌、棉花枯萎病菌的抑菌活性。结果表明,黄花蒿提取物对玉米小斑病菌的生物活性好于棉花枯萎病菌;黄花蒿叶的提取物抑菌效果最好,根的抑菌效果最差;叶的石油醚(60℃～90℃)提取物对玉米小斑病菌EC50为156.32mg/L;而叶的丙酮提取物对玉米小斑病菌的EC50为82.37mg/L。

Target-site Mechanisms Involved in Annual Bluegrass(Poa annua L.) Tolerance to Fenoxaprop-P-ethyl

Annual bluegrass （Poa annua L.） was found to be tolerant to fenoxaprop- P-ethyl as well as quizalofop-P-ethyl, haloxyfop-R-methyl, clodinafop-propargyl, fluaz- ifop-P-butyl, cyhalofop-butyl, sethoxydim and tralkoxydim, whereas it was sensitive to clethodim and tepraloxydim. The acetyI-CoA carboxylase （ACCase） IC50 values of five P. annua biotypes were 10.46 to 11.98-fold higher than the susceptible Japanese foxtail （Alopecurus japonicus Steud.）. The presence of the polymorphic lie and Leu at 1 781, which the presence of Leu at 1 781 had been reported to be in- volved in the resistance of grass weeds to ACCase inhibitors, was subsequently i- dentified in the ACCase of P. annua. Furthermore, the expression level of gene that encoding P. annua ACCase was found to be approximately 4.67 to 7.37-fold higher than A. japonicus, possibly explaining the P. annua target site tolerance to fenoxaprop-P-ethyl.

Effects of traffic pollution on the genetic structure of Poa annua L.populations

The genetic composition of Poa annua L. populations with a series of traffic pollution was studied by starch electrophoresis. Five enzyme systems were stained. The results showed that: (1) Traffic pollution can dramatically change genotypic frequencies at some loci of P. annua populations. Significant deviations from Hardy Weinberg equilibrium were observed on loci Fe 1 and Me due to the excess of heterozygotes in some populations. (2) The effective number of alleles per locus and the observed and expected heterozygosity were higher in the pollution series than in the clear control site(Botanic Park population), but the increase was not related with the pollution extent. (3) Most genetic variation was found within populations, and only 6 21% was among populations of the polluted series. Slightly higher differentiation( F ST =7 98%) was observed when the control population was included. (4) The calculated gene flow(Nm) is 2 8841 per generation. The mean of genetic identity is 0 9864 and the genetic distance average to 0 0138

Effects of Different Herbicides on the Control of Malachium aquaticum L. and Poa annua L.

In this experiment,the effects of 13 kinds of herbicides( or mixtures) on the control of two kinds of weeds were studied by pot cultivation with the broadleaf weed Malachium aquaticum L. and Gramineae weed Poa annua L. as the materials. The results showed that the herbicide MCPA-Na had the best and fastest control effect on M. aquaticum L.,and all the M. aquaticum L. died 7 d after treatment;it was followed by the other four herbicides including MCPA-Na + clethodim,MCPA-Na + quizalofop-p-ethyl,bentazon and nicosulfuron·atrazine,and all the M. aquaticum L. died 14 d after treatment. Atrazine was the best herbicide to control Gramineae weeds,followed by nicosulfuron·atrazine,and mesotrione·nicosulfuron·atrazine. The study on the application of field herbicide found that four herbicides including atrazine,mesotrione·nicosulfuron·atrazine,nicosulfuron·atrazine and bentazon had better control effect on weeds. The best herbicide for flax field was MCPA-Na + clethodim,followed by MCPA-Na and MCPA-Na + quizalofop-p-ethyl. The optimized herbicides and combinations had no harmful effects on the growth of corn and flax.

HPLC法同时测定黄花蒿水提浸膏中青蒿素、青蒿乙素、猫眼草黄素和猫眼草酚D的含量

目的建立高效液相色谱(HPLC)法同时测定黄花蒿水提浸膏中青蒿素、青蒿乙素、猫眼草黄素、猫眼草酚D的含量。方法采用Agilent ZORBAX SB-C18(250 mm×4.6 mm,5μm)色谱柱;流动相:乙腈(A)-纯水(B),梯度洗脱;体积流量:0.8 mL·min^(-1);柱温:30℃;进样量:10μL;检测波长:210 nm。结果青蒿素、青蒿乙素、猫眼草黄素和猫眼草酚D进样量分别在1.6088~16.088μg(r=0.9999)、0.0141~0.1414μg(r=1)、0.1851~1.8509μg(r=0.9999)、0.1441~1.4414μg(r=0.9999)范围内与峰面积呈良好的线性关系,平均加样回收率(n=6)分别为102.44%、97.82%、95.07%、95.55%,RSD分别为1.12%、1.44%、1.29%、1.53%。结论该方法简便准确、可靠稳定、重复性好,可用于同时测定黄花蒿水提浸膏中青蒿素、青蒿乙素、猫眼草酚D、猫眼草黄素的含量。

Effect of Artemisia annua L. as Substitute to Sulfonamides (Sodium Sulfadimerzine) on Coccidiosis and Growth Performance in Rabbits

Coccidiosis is a disease caused by protozoa of the genus Eimeria which seriously affects young rabbits. Treatment based on the use of anticoccidial drugs is increasingly ineffective due to the rapid emergence of resistant strains of coccidia and the high cost of drugs. Consumer demand for rabbit products without chemical residues led to a growing interest in the use of medicinal plants as an alternative treatment for coccidiosis. The present study was carried out during the period of August to December 2020 to assess the anticoccidial effect of hydro-ethanolic extract of leaves of Artemisia annua L., in young rabbits. The antiparasitic efficacy of Artemisia extract was tested on 15 young rabbits (whose age varied between 7 and 9 weeks) divided into 5 lots of 3 animals. The average weight of these animals was 790 g. The results of this study show that the feces samples and the weight of young rabbits before administration of the treatment and the coprological examination (every 7 days for 4 weeks) show a fecal excretion reduction rate (FECRT) of 55.13% in the lot treated by sulfonamide. On the other hand, in animals received treatments extract of the leaves of Artemisia annua L., the average FECRT is evaluated at 69.64%, 79.22%, and 96.36% for respective doses of 400, 800 and 1200 mg/kg bodyweight and proves their anticoccidial effect. Furthermore, the variation in mean Eggs Per Gram (EPG) of coccidia and the average weekly weight gain (AWWG) of each lot were significant in the lots treated with hydro-ethanolic extract (P 0.05). The greatest reductions in oocystal excretion and weight gain obtained were those of lot 5, treated at 1200 mg/kg of hydro-ethanolic leaves extract of Artemisia annua L.

黄花蒿中青蒿素含量的RP-HPLC法测定

建立黄花蒿中青蒿素的高效液相色谱测定方法,采用柱前衍生RP-HPLC法测定黄花蒿中青蒿素的含量,采用ZORBAXXDB-C18(4.6mm×150mm,5μm)色谱柱,甲醇-0.01mol/L醋酸钠-醋酸缓冲液(pH5.8,体积比62∶38)为流动相;检测波长:260nm;流速:0.8mL/min;柱温:30℃。对广西、沈阳、北京、郑州、苏州和杭州等不同产地的野生黄花蒿样品、以及同一产地不同采集时间黄花蒿样品进行检测,结果表明不同地区青蒿素含量差异很大,同一地区不同采集时间黄花蒿样品的青蒿素含量也有差异。该法准确可靠、重现性好,能准确地反映青蒿素含量的检测。

HPLC-ELSD法测定青蒿中青蒿素的含量

目的：首次建立高效液相色谱-蒸发光散射检测器测定青蒿素含量的方法，并对大量的青蒿样品进行含量测定。方法：采用蒸发光散射检测器（ELSD）对青蒿药材中的青蒿素进行HPLC分析，色谱柱：迪马公司Diarnonsil C18（250×4．6mm，5μm）；流动相：甲醇-水（75：25）；流速：1ml／min；蒸发光散射检测器漂移管温度40℃，载气压力3．5bar，放大系数（gain）为9；进样体积20μl。结果：青蒿索在1～5μg范围线性关系良好，回收率为99．33％（RSD=1．97％）。结论：本法具有良好的精密度和重现性，结果准确可靠，可作为青蒿药材及青蒿素类产品的质量控制方法。不同产地、不同采收期青蒿药材中青蒿素的含量测定为青蒿药材的收购及栽培等提供依据。

RP-HPLC法测定青蒿中青蒿素的含量

目的建立青蒿(黄花蒿)干叶中青蒿素的含量测定方法,为甲醇提取青蒿素的工业化生产工艺提供依据。方法采用正交试验设计,优选测定条件,采用Shimadzu Shim-pack VP-ODS C18柱(250 mm×4.6 mm,5μm),流动相为乙腈-水(55∶45),流速为1.0 mL.min-1,检测波长为210 nm。结果最佳的含量测定条件为加40倍量甲醇,超声提取3次,每次50 min。青蒿素在1～40μg范围内与峰面积呈良好的线性关系(r=0.99998),平均加样回收率为99.8%,RSD为1.0%(n=6)。结论该方法简便、准确,适用于黄花蒿中青蒿素的含量测定。

黄花蒿中青蒿素的柱前衍生HPLC-UV分析

研究衍生化条件对HPLC-UV法测定黄花蒿青蒿素含量的影响。建立了黄花蒿青蒿素的柱前衍生HPLC-UV测定方法:即青蒿素标准品溶液(或黄花蒿提取浸膏)以体积分数为95%乙醇补充(溶解)至15.0mL,再加入质量分数为0.2%的NaOH8.0mL,45℃水浴30min后冷却至室温,加入0.08mol/L的HAc10mL摇匀,以95%乙醇定容至50mL容量瓶中;HPLC分离,260nm紫外检测。所得方法检测线性范围为0～200mg/L,相对标准偏差(RSD)值为1.40%,平均回收率(n=5)为100.1%。用该方法测定了黑龙江引种的黄花蒿提取物样品,稳定性RSD值为4.35%,重现性RSD值为4.96%。证明该方法简单、稳定、重现性好。

衍生化条件的优化及不同产地青蒿中青蒿素含量的柱前衍生化-HPLC法测定

Derivation conditions of pre-column derivation-HPLC method used for determinating artemisinin content were compared and selected,and artemisinin content in above-ground part of Artemisia annua L.from seventeen locations was compared by optimal pre-column derivation-HPLC method.The optimal derivation condition is selected via comparing of 0.2% NaOH solution addition(3,4,5,6 and 7 mL),derivation temperature(30 ℃,35 ℃,40 ℃,45 ℃ and 50 ℃) and derivation time(0,2,5,10,20,40 and 60 min) during derivation process.The optimal derivation condition is adding 5 mL 0.2% NaOH solution,then insulating in 40 ℃ water bath for 10 min.Differences of artemisinin content in A.annua from seventeen locations are obvious,the general trend is that artemisinin content decreases gradually with location from south to north,in which artemisinin content in A.annua from Youyang of Chongqing is the highest with a value of 7.08 mg·g-1.The method is simple and accurate with good reproducibility,and can be used to determinate artemisinin content in medicinal material of A.annua.

黄花蒿提取物生产工艺条件及其对乳中共轭亚油酸(CLA)生成的调控作用

黄花蒿乙醇提取物含有多种营养活性物质,主要包括萜类、倍半萜类、芳香族类、脂肪酸类、甾体类、黄酮类、生物碱类等化合物。反刍动物日粮中添加黄花蒿乙醇提取物能够增加其乳中共轭亚油酸(CLA)含量。综述了黄花蒿乙醇提取物的主要营养活性物质、提取工艺条件、营养活性物质的检测方法及黄花蒿提取物在动物日粮中的适宜添加量,旨在为生产CLA乳提供参考。

UPLC-MS/MS法检测青蒿中青蒿素的含量

采用UPLC-MS/MS法检测青蒿中青蒿素的含量,色谱柱Waters ACQUITY BEH C18(50 mm×2.5mm,1.9μm),以甲醇和水梯度洗脱,甲醇-水体积比为40%(0 min)-70%(4.5 min)-40%(5.0 min)-40%(5.5 min),流速0.30 mL/min,柱温30℃,进样量3.0μL,离子模式(ESI+)分析,青蒿素母离子为m/z=283.3。子离子m/z=151.2,m/z=209.4。结果表明,在上述条件下,青蒿素与杂质分离完全,无明显干扰,检出限达到0.174 ng,平均回收率为99.38%,精密度良好,青蒿中青蒿素含量为(1 045.19±27.55)mg/kg,该法适用于青蒿素分析。

黄花蒿bHLH转录因子基因家族鉴定及光调控分析

目的:基于黄花蒿全基因组及转录组数据,进行AabHLH基因家族生物信息学分析及在不同光处理下的基因表达分析。方法:基于基因隐马科夫模型(HMM)对AabHLH基因进行鉴定,利用Pfam、GSDS、MEME等在线分析软件对基因结构、分子进化及保守结构域等进行分析。结果:全基因组水平共鉴定出99条黄花蒿AabHLH基因,包含49对复制基因对;基于系统发育树将其分为13个亚族,所有亚族AabHLH蛋白亚细胞定位于细胞核且均为亲水性蛋白,同一亚族基因具有相似的外显子-内含子结构及保守基序。基因表达模式分析发现,AabHLH基因在不同水平上响应蓝光、红光、远红光、白光调控,推测其中6个亚族基因为光照条件下,7个亚族基因为非光照条件下青蒿素合成途径调控基因。结论:在全基因组水平鉴定了AabHLH基因,为深度解析AabHLH基因功能及其对光调控下青蒿素生物合成调控机制提供参考。

青蒿UPLC指纹图谱的建立与多种成分含量测定

目的:建立30批青蒿样品的UPLC指纹图谱及有机酸类、黄酮类、香豆素类等成分的含量测定方法。方法:采用Agilent Eclipise Plus C18(50 mm×2.1 mm,1.8μm)色谱柱;以0.1%磷酸溶液-甲醇为流动相,梯度洗脱;建立指纹图谱;并对青蒿中有机酸类、黄酮类、香豆素类等9个成分进行含量测定。结果:建立了青蒿UPLC指纹图谱及同时测定多成分含量的方法。9种成分在测定范围内线性关系良好,不同产地的青蒿药材质量存在一定差异。江苏盱眙的青蒿综合评价质量最优。结论:本实验建立的方法准确可靠,可同时测定青蒿中东莨菪内酯、滨蒿内酯、异绿原酸A、异绿原酸B、异绿原酸C、木犀草素、芹菜素、猫眼草黄素、艾黄素的含量。

HPLC-ELSD法测定黄花蒿提取物中青蒿素

目的 建立HPLC-ELSD法对黄花蒿提取物中青蒿素的测定方法。方法 采用HPLC-ELSD法。色谱柱：Inertsil ODS-3（150mm×4．6mm，5μm）；流动相为乙腈-水（69：31，用三氟醋酸调pH至3）；体积流量：1．0mL／min；检测器漂移管温度：45C；载气体积流量：1．0L／min；分流：开；柱温：25℃。结果 青蒿素在0．5893～5．8935mg／mL时面积的常用对数值与其质量浓度的常用对数值线性关系良好（r=0．989）；样品平均回收率为98．71％（RSD为0．42％，n=6）。结论 该方法灵敏、准确可靠，可作为黄花蒿提取物中青蒿素的定量方法。

基于UHPLC-MS/MS的药用植物内源性激素含量检测方法及黄花蒿不同器官激素的测定

目的激素是调控植物活性次生代谢产物合成途径的核心调控因子,内源激素结构多样、含量极低,检测难度高,缺乏高效、准确的检测方法。本研究利用超高效液相色谱串联质谱(UHPLC-MS/MS)技术构建了对黄花蒿中茉莉酸、细胞分裂素、赤霉素、生长素、脱落酸等多类植物内源性激素高效定量检测方法。方法使用Agilent Poroshell 120 EC-C_(18)(2.7μm,3.0×150 mm)色谱柱,流动相为0.05%甲酸水-乙腈溶液,梯度洗脱,采用多反应监测模式验证方法可行性,并对黄花蒿不同器官中植物激素进行定量分析。结果本方法中检测的16种植物激素均呈良好线性,加样回收率、精密度和稳定性均符合植物样品的测定要求。利用此方法测定青蒿不同器官的激素含量,结果表明脱落酸、茉莉酸、茉莉酸甲酯均在叶中含量较高,赤霉素和生长素在根部含量高,细胞分裂素在叶和茎中含量高。结论青蒿不同器官的激素测定结果可进一步印证脱落酸、茉莉酸、茉莉酸甲酯在调控青蒿素合成中起着重要作用的相关报道,同时表明该方法检测性良好。本文构建的植物内源性激素的检测方法为进一步探究植物生长发育、抵御外界胁迫的相关机制及次生代谢产物的合成等方面提供了手段。

黄花蒿对大蒜化感作用研究

为探究黄花蒿对大蒜鳞茎萌发及其幼苗生长的化感作用,用水提取黄花蒿叶、根、根际土壤,获取不同质量浓度的浸提液,用于处理大蒜鳞茎,测定大蒜鳞茎萌发率及其幼苗生长状况和叶绿素含量等指标。结果表明:随着黄花蒿水浸液浓度的升高,黄花蒿水浸液对大蒜鳞茎萌发率及其幼苗苗高、根长、叶绿素a和b含量、叶绿素含量、过氧化物酶活性、植株鲜质量、植株高的抑制作用不断增大,对大蒜幼苗丙二醛含量的促进作用不断增强;黄花蒿水浸液通过抑制大蒜鳞茎萌发率及其幼苗过氧化物酶活性、增加膜脂过氧化和逆境伤害、抑制叶绿素a、b和叶绿素的生成,从而抑制植株生长,表现出化感作用,且化感效应随着黄花蒿水浸液浓度的升高而增大。本研究为农作物和药用植物栽培及植物间化感作用研究提供参考。

银扇草果实的脂肪酸分析研究

银扇草(Lunaria annua L.)属十字花科银扇草属,是一种含神经酸的二年生草本植物,其种子富含油脂,具有较高的生物与医药价值。为探明银扇草果实发育过程中脂肪酸的种类和含量变化特征,以及超长链脂肪酸特别是神经酸的合成特点,本研究使用气相色谱-质谱联用技术比较分析了含神经酸的银扇草、琉璃苣和蒜头果果实的脂肪酸组分及含量。结果表明,银扇草果实发育过程中共有10种脂肪酸参与了油脂代谢,种子中有7种,果皮中有8种,多为长链及超长链不饱和脂肪酸。银扇草种子内的脂肪酸主要由软脂酸、硬脂酸、油酸、亚油酸、亚麻酸、芥酸和神经酸组成;软脂酸、硬脂酸、亚油酸和亚麻酸的含量均随种子发育逐渐减少;油酸、芥酸和神经酸随种子发育逐渐增加,并在花后25~35 d大量累积;果皮中不含芥酸和神经酸,但有山嵛酸、木质素酸和蜡酸。银扇草、琉璃苣和蒜头果种子中均含芥酸和神经酸,木本植物蒜头中果含量最高,银扇草次之,琉璃苣含量最低,三者种子中神经酸的含量与芥酸含量有明显的正相关关系。本研究结果阐明了银扇草果实发育过程中脂肪酸代谢变化特点及神经酸积累特性,为充分利用银扇草种质资源和拓展神经酸的获取途径提供了新思路。

不同采收期和种植地貌青蒿的品质分析

目的:比较不同采收期喀斯特山地和传统平地上种植青蒿的表型特征和有效成分含量变化,确定青蒿的最适种植环境及最佳采收期。方法:采用直接测量法、高效液相色谱法和比色法,分别测定2种地貌上种植的5个采收日期青蒿的表型特征,以及青蒿素、青蒿酸、总黄酮和总多酚含量。结果:同一采收日期,在喀斯特山地上种植的青蒿呈全株矮且宽、根系稀疏且为浅根系的生长状态,而在传统平地种植的青蒿呈枝繁叶茂、株高茎壮、根系茂密且为深根系的生长状态;传统平地种植的青蒿总黄酮、总多酚、青蒿素、青蒿酸含量高于喀斯特山地种植的青蒿。喀斯特山地和传统平地上种植青蒿的总黄酮、青蒿素、青蒿酸含量均在7月末最高,总多酚含量的变化趋势与青蒿表型特征的变化趋势相似。结论:传统平地上种植的青蒿表型特征及青蒿素等成分含量的最高值均优于喀斯特山地上种植的青蒿,7月下旬为青蒿的最佳采收期。