Extraction and Purification Process Optimization and Antioxidant Activity in vitro of Flavonoids in Amaranthus caudatus L.

[Objectives]To explore the optimal extraction and purification process of the flavonoids in Amaranthus caudatus L.and to study the antioxidant activity in vitro of the flavonoids in A.caudatus.[Methods]Taking A.caudatus as the raw material,flavonoids were extracted by alcohol extraction method,and AB-8 macroporous adsorption resin was selected for purification.The hydroxyl radical scavenging ability,DPPH radical scavenging ability,and O^2-radical scavenging ability were used as evaluation indicators,to explore the antioxidant activity in vitro of the flavonoids in A.caudatus.[Results]The optimal extraction process conditions of flavonoids in A.caudatus are:liquid-to-material ratio 40:1,extraction temperature 60℃,ethanol concentration 60%,ultrasonic power 320 W,extraction time 50 min.Under these conditions,the extraction yield of flavonoids in A.caudatus is(1.35±0.01)%.The optimal purification process conditions of flavonoids in A.caudatus are 2.5 g AB-8 macroporous adsorption resin,sample volume 5 mL,mass concentration of adsorption solution 1.60 mg/mL,pH value of adsorption solution 3.0,sample flow rate 3 BV/h,ethanol concentration in desorption process is 70%and the desorption flow rate is 3 BV/h.Under these conditions,the recovery rate reaches 88.35%±0.68%.[Conclusions]A.caudatus has a high content of flavonoids and has excellent free radical scavenging ability in vitro.This study is intended to provide important technical support for the research of flavonoid activity of A.caudatus and the development of functional products.

Dormancy of Amaranthus retroflexus L. Seeds and Physiological Response Seedlings to Acifluorfen Sodium

Amaranthus retroflexus L. is a serious and widespread malignant weed in soybean fields in Heilongjiang Province. Exploring the dormancy characteristics of A. retroflexus L. seeds and the physiological response of its seedlings to acifluorfen sodium can provide a basis for further researches on its resistance mechanism. Using newly harvested and stored A. retroflexus L. seeds for one year as experimental materials, the effects of different concentrations of HCl, NaOH, water temperature, gibberellic acid(GA) and polyethylene glycol(PEG) on the dormancy and germination of A. retroflexus L. seeds were studied. The sensitivity of A. retroflexus L.to acifluorfen sodium was determined using bioassay. The effects on leaf chlorophyll content and target enzyme activity were studied at a normal dosage of 360 g a.i. hm^(-2) and a doubling dosage of 720 g a.i. hm^(-2) of acifluorfen sodium. Newly harvested seeds exhibiting dormancy were soaked in water of various temperatures and in different concentrations of NaOH and HCl, which were ineffective in breaking the seed dormancy. GA could break seed dormancy, and the highest seed germination rate reached 93.33% when they were soaked at 3 000 mg·L^(-1) for 72 h and 4 000 mg·L^(-1) for 48 h. The drought stress was simulated with a 15%-25% polyethylene glycol solution, which had no significant effect on the seed germination rate. The GR_(50) value of acifluorfen sodium for A. retroflexus L. was 705.7 g a.i. hm^(-2), which was 1.96 times the recommended dose in the field. After the application of different doses of acifluorfen sodium, the chlorophyll content of A. retroflexus L. reached its minimum value 3 days after treatment(DAT), and then gradually increased. The activity of the target enzyme protoporphyrinogen oxidase(PPO) reached the highest value at 7 DAT under different dosages, and gradually returned to normal levels thereafter. Soaking with gibberellin was an effective method to break seed dormancy. A. retroflexus L. seeds had certain drought resistance during the germination process. A. retroflexus L. was not sensitive to acifluorfen sodium and acifluorfen sodium ether, and could not effectively inhibit the PPO activity, indicating that A. retroflexus L. had target resistance to acifluorfen sodium.

EDTA和柠檬酸对反枝苋(Amaranthus retroflexus L.)Cu迁移富集影响研究

为明确反枝苋(Amaranthus retroflexus L.)对Cu的富集能力及EDTA与柠檬酸(CA)对Cu迁移的影响,采用盆栽试验,在0、100、200、400、800 mg·kg-1 Cu处理质量分数及2.5 mmol·kg-1 EDTA和柠檬酸施加条件下,测定了根际土、非根际土Cu不同形态含量及反枝苋组织Cu富集特征。结果表明:Cu处理质量分数小于400 mg·kg-1时,重金属Cu组织富集特征为根>叶>茎,Cu处理质量分数在400 mg·kg-1时,富集特征为根>茎>叶,Cu各处理组BCF与TF均小于1,不具备Cu超富集植物特征;EDTA、柠檬酸均可促进反枝苋对土壤中Cu的富集,并强化Cu由根系向地上部分的转移。EDTA的促进作用远高于柠檬酸,在Cu 800 mg·kg-1,施EDTA处理组反枝苋地上部分Cu可达3 513 mg·kg-1。施加EDTA可增加根际土全Cu含量,显著减少根际土中Fe-Mn态Cu含量,增加Exch态,促使Fe-Mn态向Exch态的转化;柠檬酸处理的根际土中则以Fe-Mn态向Carb态转化为主。由此可见,在EDTA强化修复下,反枝苋富集Cu的能力明显增加,而根际土中Cu全量的增加,是Cu-EDTA迁移活性极大增加与反枝苋防御性Cu阻隔共同作用的结果。

Anti-diabetic and anti-cholesterolemic activity of methanol extracts of three species of Amaranthus

Objective:To investigate the anti-diabetic and anti-cholesterolemic activity of methanol extracts of leaves of Amaranthus caudatus,Amaranthus spinosus and Amaranthus viridis in normal and streptozotocin(STZ) induced diabetic rats.Methods:In this study,the anti-diabetic and anti-cholesterolemic activity of methanol extracts of leaves of all three plants was evaluated by using normal and STZ induced diabetic rots at a dose of 200 mg/kg and 400 mg/kg p.o.daily for 21 days.Blood glucose levels and body weight were monitored at specific intervals,and different biochemical parameters,serum cholesterol,serum triglyceride,high density lipoprotein, low density lipoprotein and very low density lipoprotein were also assessed in the experimental animals.Histology of pancreas was performed.Results:It was found that all the three plants at 400 mg/kg dose showed significant anti-diabetic and anti-cholesterolemic activity(P<0.01),while at 200 mg/kg dose less significant anti-diabetic activity(P<0.05) was observed.Concluslons: Methanol extracts of Amaranthus caudatus,Amaranlhus spinosus and Amaranthus viridis showed significant anti-diabetic and anti-cholesterolemic activity,which provides the scientific proof for their traditional claims.

不同积累型苋菜(Amaranthus mangostanus L.)镉吸收转运特征差异性研究

【目的】比较高积累型苋菜品种Tianxingmi与低积累型苋菜品种Zibeixian在Cd胁迫下Cd吸收转运特征差异性,揭示苋菜品种Tianxingmi高积累镉的机理。【方法】采用水培试验,添加代谢抑制剂来研究两品种苋菜根系吸收Cd的主要途径。分别在30μmol/L Cd Cl2处理4 h、8 h、16 h、1 d和2 d采样测定,采用非损伤微测技术(Non-invasive micro-test technique,NMT),进行植物活体动态测试,调查、测量两品种苋菜根系Cd^(2+)离子流特征,比较了两品种对代谢抑制剂的反应。【结果】在30μmol/L Cd Cl2处理1 d后,Tianxingmi生物量达到最大值5.90 g/plant,是Zibeixian生物量的二倍;Tianxingmi根、茎和叶中Cd浓度分别为609、254和62.3mg/kg,分别是Zibeixian的1.4倍、1.9倍和1.6倍,地上部和全株的Cd累积量分别高达602.0、1308μg/plant;Tianxingmi富集系数(BCF)与转运系数(TF)分别为Zibeixian的2.1倍和1.5倍,这些结果两品种均表现出显著性差异(P<0.05)。NMT技术测定两品种根系Cd^(2+)离子流,通过扫描位点测定发现,距根尖0~300μm范围内两品种苋菜根系Cd^(2+)内流最强且差别最大,并在此做定点位点测定,发现Tianxingmi根系Cd^(2+)内流是Zibeixian根系Cd^(2+)内流的3.75倍,说明了两品种的富集特征与NMT结果一致。添加代谢抑制剂处理显著降低了高积累型Tianxingmi各器官中的Cd浓度,富集系数(BCF)与转运系数(TF),以及根系Cd^(2+)内流(P<0.05),主动吸收特征明显,而对低积累型Zibeixian的相关生理指标影响不大。【结论】两苋菜品种Cd吸收转运特征均表现出显著性差异。Tianxingmi具有更强的Cd吸收和向地上部转运与累积能力,并且根系对Cd的吸收与转运是主动需能的过程,共质体途径在Cd进入Tianxingmi根系并向地上部运输的过程中起着主要作用。

An ABAGE-Like Metabolite of Botrytis cinerea Isolate BC4 Inhibited the Growth of Hypocotyls and Roots of Amaranthus retroflexus Seedlings

A metabolite, which had an inhibitory effect on plant growth, was isolated from cultural filtrate of Botrytis cinerea isolate BC4 by column chromatography on silica gel and preparative HPLC. Its structure was determined from HPLC-ESI MS, GC, IR, ^1H NMR and ^13C NMR spectral data, as well as chemical hydrolysis. The results showed that this metabolite was quite similar to abscisic acid-β-D-glucopyranosyl ester（ABAGE） in structure. The inhibitory effect of the ABAGE-like metabolite on plant growth was investigated using a weed Amaranthus retroflexus L. as a bioassay material. The results showed that it inhibited hypocotyls and roots growth of A. retroflexus （Amaranthus retroflexus L.） seedlings when its concentration was over 0.5 μM. The concentrations for 50% inhibition of hypocotyls and roots growth of A. retroflexus seedlings were 2.8 and 1.4 μM, respectively.

苋菜LAC基因家族成员全基因组鉴定与表达分析

漆酶作为生物体内一类重要的铜蓝蛋白,起着调控植物生长、抵抗逆境的作用。基于苋菜基因组及转录组数据,对苋菜漆酶基因家族成员(AtrLAC)进行鉴定,利用在线软件进行生物信息学分析,并根据转录组TPM值进行家族成员相对表达分析。结果表明,在苋菜基因组中共筛选获得19个LAC基因;AtrLAC家族蛋白长度范围在118~1191 aa之间,分子量为13.05~135.10 kD;理论等电点范围为6.49~9.85,多数为亲水呈碱性稳定的蛋白;亚细胞定位预测显示多数在细胞膜外或质膜上。系统进化树分析将苋菜19个LAC成员和拟南芥的17个LAC成员聚为6个类群。TPM值分析显示,AtrLACs在不同处理下的表达量存在差异,其中AtrLAC7基因可能在甜菜色素合成、响应激素调控及光条件下发挥重要作用。

Pain management in mice using methanol extracts of three plants belongs to family Amaranthaceae

Objective:To investigate the analgesic activity of methanolic extract of Amaranthus viridis(A. viridis),Amaranthus caudatus(A.caudatus) and Amaranthus spinosus(A.spinosus).Methods:In this study,the analgesic activity of methanol extracts of all three plants at doses of 200 and 400 mg/kg were investigated by acetic acid-induced writhings test,hot plate test and tail immersion test for mice.Results:It was found that all the three plants showed significant pain management effect(P【0.01) at a dose of 400 mg/kg,but showed a less significant effect at a dose of 20 mg/kg in the entire tests used for evaluation of analgesic activities(P【0.05).Conclusions:Methanol extracts of A.viridis,A.caudatus and A.spinosus show potent analgesic activities,and this study provides the scientific proof for their traditional claims.

An ABAGS-Like Metabolite of Botrytis cinerea Isolate BC4 and Its Inhibitory Activity to Seed Germination of Weeds

Aimed at finding out natural compounds to kill weeds, a plant pathogenic fungus, Botrytis cinerea isolate BC4, was chosen as a source. A significantly polar metabolite, which was different in chromatographic behavior from ABAGE-like metabolite, was isolated from cultural filtrate of the fungus by column chromatography on silica gel, preparative LC, TLC and HPLC. The structure of the metabolite was determined by HPLC-ESI MS, g.l.c, IR, ^1H NMR and hydrolysis. The results showed that this metabolite was quite similar with ABAGS. The bioassay showed the metabolite had inhibition to seed germination of both broadleaf weeds [Amaranthus retroflerus L. and Chenopodium album L.] and gramineae weeds [Digitaria sanguinalis L. Scop and Echinochloa crusgalii L. Beauv]. It also had inhibitory activity to the seedling growth of broadleaf weeds. The bioassay, using Amaranthus retroflexus L. as an indicator, showed that the lowest concentration of the metabolite to inhibit root＋sprout growth of A. retroflexus was 0.6 μM, and the concentration for 50% inhibition was 1.3μM.

Influence of Adjuvants on the Control of Glyphosate-Resistant Canada Fleabane and Waterhemp in Corn with Tolpyralate

Tolpyralate is a new benzoylpyrazole herbicide for weed management in corn. It is recommended to be co-applied with atrazine along with the adjuvants methylated seed oil concentrate (MSO) plus an ammonium nitrogen fertilizer, such as urea ammonium nitrate (UAN). Two studies were conducted on glyphosate-resistant (GR) Canada fleabane and GR waterhemp to determine if an additional adjuvant is still required when tolpyralate plus atrazine are tankmixed with a commercial glyphosate formulation (Roundup WeatherMAX&reg;) in corn. Trials were conducted over a two-year period (2018-19) on farms in south western Ontario with confirmed GR populations. When co-applied with Roundup WeatherMAX&reg;, the addition of MSO to tolpyralate + atrazine increased control of GR waterhemp 9%;however, there was no increase in GR Canada fleabane control from the addition of additional adjuvants. At 8 WAA, all treatments provided > 91% and > 84% control of GR waterhemp and GR Canada fleabane, respectively. This study concludes that the addition of Roundup WeatherMAX&reg;to tolpyralate plus atrazine improves the control of GR waterhemp and GR Canada fleabane in corn.

反枝苋对氟磺胺草醚的抗性机制代谢组学研究

二苯醚类除草剂氟磺胺草醚是防除大豆田杂草反枝苋Amaranthus retroflexus L.的主要除草剂,但黑龙江省大豆田反枝苋对氟磺胺草醚的抗性越来越强,严重影响大豆产量。目前尚无关于反枝苋对氟磺胺草醚抗性机制的代谢组学研究。本研究基于未处理(CK)以及氟磺胺草醚处理后反枝苋敏感种群(SY)、抗性种群(RY)的叶组织的代谢谱,进行主成分分析(PCA)和正交偏最小二乘法判别分析(OPLS-DA),发现RY组代谢分子表型更贴近于CK组。京都基因与基因组百科全书(KEGG)富集分析和拓扑分析结果发现,氟磺胺草醚的主要作用通路有3条,分别是biosynthesis of amino acids (氨基酸的生物合成)、arginine biosynthesis (精氨酸生物合成)和2-oxocarboxylic acid metabolism (2-氧代羧酸代谢)。CK vs RY和SY vs RY有1条共有的差异通路——degradation of flavonoids (类黄酮的降解),因此,类黄酮的降解可能是RY组抗性的特异性通路。通过支持向量机(SVM)分析进一步发现,4-hydroxy-2-oxoglutaric acid (4-羟基2-氧化戊二酸)、citrulline (瓜氨酸)和L-ornithine (L-鸟氨酸)是氟磺胺草醚处理后反枝苋产生抗药性的关键代谢物。

反枝苋对氟磺胺草醚的抗性水平及分子机制

[目的]本研究旨在明确黑龙江省大豆田反枝苋(Amaranthus retroflexus L.)种群对氟磺胺草醚的抗药性水平,并研究其产生抗性及多抗性的分子机制。[方法]利用琼脂法快速测定采自黑龙江省嫩江县的3个反枝苋种群对氟磺胺草醚的抗性水平。通过盆栽法筛选出抗氟磺胺草醚/噻吩磺隆的反枝苋植株,并进行原卟啉原氧化酶(protoporphyrinogen Ⅸ oxidase, PPO)和乙酰乳酸合酶(acetolactate synthase, ALS)的靶标基因测序。[结果]相比于敏感种群,黑龙江省3个种群对氟磺胺草醚的相对抗性倍数(resistance index, RI)分别为83.6、83.8和93.4;位于反枝苋PPO2高度保守区域编码128位氨基酸的核苷酸由AGG突变为GGG,导致精氨酸被替换为甘氨酸(Arg-128-Gly)。3个抗氟磺胺草醚的反枝苋种群均对ALS抑制剂类除草剂产生多抗性。对als基因测序后发现,与敏感种群相比,各抗性种群10个单株的ALS保守区域均发生多个不同位点(Ala-205-Val、Asp-376-Glu、Trp-574-Leu和Ser-653-Thr)及组合的氨基酸取代方式,各种群突变位点和杂合度多样。[结论]PPO2的Arg-128-Gly氨基酸取代可能是黑龙江3个反枝苋种群对氟磺胺草醚产生抗性的主要原因,3个种群同时对ALS抑制剂类除草剂存在靶标位点突变导致的多抗性。

Effects of G protein and cGMP on phytochrome-mediated amaranthin synthesis in Amaranthus caudatus seedlings

The effects of G protein and cGMP on phytochrome mediated amaranthin biosynthesis in Amaranthus caudatus seedlings were studied. It was shown that G protein agonist cholera toxin induced amaranthin synthesis in darkness, whereas G protein antagonist pertussis toxin inhibited red light induced amaranthin synthesis. Amaranthin synthesis was also induced by exogenous cGMP, while the amaranthin biosynthesis induced by cholera toxin, red light and exogenous cGMP was inhibited by genistein. LY 83583, an inhibitor of guanylyl cyclase, inhibited the amaranthin synthesis induced both by red light and cholera toxin, while it was not able to inhibit the amaranthin synthesis induced by exogenous cGMP. These results suggest that G protein, guanylyl cyclase and cGMP were the candidates in phytochrome signal transduction chain for red light induced amaranthin biosynthesis and the red light signal transduction chain might be as follows: red light→phytochrome→G protein→guanylyl cyclase→cGMP.

基于DNA条形码鉴别青葙子及其混伪品绿穗苋种子

目的:基于DNA条形码对青葙子、绿穗苋种子进行鉴定。方法:基于ITS2、psbA-trnH、matK、rbcL常用DNA条形码的通用引物对样品进行扩增、测序,并从NCBI数据库下载ITS2、psbA-trnH、matK、rbcL碱基序列。利用MEGA 11.0软件构建NJ系统发育树、计算遗传距离及比较种间差异,并基于ITS2序列构建二级结构。结果:基于上述序列构建的NJ树均独立聚为一支,其中ITS2的种间遗传距离最大,基于ITS2构建的二级结构具有明显差异。结论:基于DNA条形码ITS2碱基序列及其二级结构可准确鉴别出青葙子及其混伪品绿穗苋种子,psbA-trnH碱基序列可以作为辅助鉴别序列。

硒和丛枝菌根真菌处理对苋菜生长及硒积累的影响

【目的】探明丛枝菌根真菌(Arbuscular mycorrhizal fungi,AMF)在苋菜富硒栽培中的生物效应,综合评价AMF对不同硒形态处理下苋菜生长和硒积累的影响。【方法】采用盆栽试验,分析了土施外源硒和接种AMF对苋菜光合作用、生长状况、苋菜红素以及硒积累的影响。【结果】硒酸钠、亚硒酸钠和L-硒代半胱氨酸处理均降低了苋菜红素含量,但对菌根侵染率和菌根依赖性均无显著影响,同时显著提高了苋菜叶片叶绿素a、b含量、蒸腾速率(T_(r))、气孔导度(G_(s))、净光合速率(P_n)和胞间CO_(2)浓度(C_(i)),其中L-硒代半胱氨酸处理效果最佳,说明外源硒处理对苋菜的生长有一定的正效应。另外,接种AMF对不同硒处理下苋菜的生长也有促进作用,且提高了叶片苋菜红素含量,同时增强了光合能力和根茎叶硒含量的积累。【结论】接种AMF和外源硒处理均能提高苋菜的光合能力和硒积累能力,从而促进苋菜生长和提高苋菜营养品质,两者交互处理可作为提高蔬菜富硒栽培的有效措施。

不同除草配方对大豆田抗性反枝苋防除效果的研究

为探索黑龙江省北部大豆主产区抗性反枝苋最佳防治方法,文章优选4种苗前封闭除草剂及4种苗后茎叶除草剂进行田间防效试验,分析不同药剂组合对反枝苋的防除效果。结果表明,苗前封闭处理,90%乙草胺EC 1800 g a.i./hm^(2)和30%丙炔氟草胺SC 51 g a.i./hm^(2)对反枝苋防效较高,施药45 d后,反枝苋防效为87.79%;苗后茎叶处理,250 g/L氟磺胺草醚AS(科坦见露灵)750 g a.i./hm^(2)+助剂(伴豆一生)2000 m L/hm^(2)对反枝苋的防效较高,且持效期长;苗前封闭结合苗后茎叶处理,90%乙草胺EC 1800 g a.i./hm^(2)+30%丙炔氟草胺SC 51 g a.i./hm^(2)+250 g/L氟磺胺草醚AS 500 g a.i./hm^(2)+480 g/L灭草松AS 1680 g a.i./hm^(2)+助剂(迪增)225 m L/hm^(2)对反枝苋的防效达93%,且持效期长。大豆田抗性反枝苋防控方法中,苗前封闭处理防效显著高于苗后茎叶处理。生产中建议以苗前封闭除草为主,结合苗后化学除草、轮作等措施综合防治反枝苋。

Effect of calcium on phytochrome-mediated synthesis of amaranthin in Amaranthus caudatus seedlings

Light plays a critical role in the regulation of plant morphogenetic and physiologicalresponses. As one of the extracellular signals, light signa1 is first received by the plantcells, then it affects the physiological and biochemical status of the cells and the expressionof genetic characters through signal transduction. Tong has pointed out that until recentlythere had been little progress in the intersteps between phytochrome activation and

鸡粪和垃圾有机肥对苋菜生长及土壤重金属积累的影响

采用盆栽试验,在红壤和潮土中分别施入土壤重量5%、10%和15%的鸡粪或垃圾有机肥,研究其对苋菜（Amaranthustricoclor L.）生长、土壤化学性状及重金属含量的影响。结果表明,与对照相比,施入鸡粪或垃圾有机肥显著增加苋菜的鲜重、主根长和株高;土壤有机质、EC值、速效磷和速效钾含量均大幅增加,但土壤全氮含量无明显变化。与对照相比,潮土中施用两类有机肥使苋菜植株Cu和Zn含量分别增加26.3%~36.0%和1.2%~20.3%,但未超出国家食品卫生标准对Cu和Zn的允许含量;Cd、Cr和Pb含量都较对照降低,没有出现积累现象。红壤施用两类有机肥,苋菜植株中Zn、Cd和Pb分别下降42.7%~59.9%、0~48.9%和4.1%~71.3%,达到显著水平。有机肥的施入量为5%时,两类土壤中重金属都没有出现明显积累;当施入量为10%和15%时,两类土壤的Cu、Cd、Cr和Pb含量显著增加,出现明显积累趋势,其中除了施用鸡粪使土壤有效Cu含量下降外,两种有机肥均增加了两类土壤中重金属Zn、Cd、Cr和Pb的有效态含量。试验结果提示,从土壤培肥与环境质量安全综合考虑,有机肥的用量应控制在一个适宜范围内。

籽粒苋对土壤中镉的耐性和积累特征

籽粒苋是一种生物量大、易栽培的镉(Cd)富集植物,具有作为Cd污染土壤修复植物的潜力.比较了两个基因型籽粒苋(K112和R104)的耐Cd性特征,土壤Cd污染浓度范围是0～50mgkg-1,生长60d收获.结果表明,籽粒苋K112(Amaranthus hypochondriacus L. Cv.‘K112’)与R104(A. hypochondriacus L. Cv.‘R104’)生物量随土壤中Cd浓度的增加而逐步下降,在土壤中Cd浓度<16mgkg-1时,其生物量积累没有受到明显的影响.在本试验最高Cd浓度条件(50mgkg-1)下仍可生长,但生物量显著下降.两种籽粒苋叶中Cd含量随土壤中Cd浓度增加而快速上升,在土壤Cd浓度为16mgkg-1时,叶内Cd浓度分别达120.63和109.96mgkg-1(DW),达到超富集植物的临界标准.Cd在植物体内的分布特征为叶>根>茎.籽粒苋两个品种相比,K112吸收Cd的能力大于R104,尤其是在高Cd浓度时,两种籽粒苋对Cd的绝对提取量相似,并随土壤Cd浓度的上升而快速增加.籽粒苋K112和R104对土壤中的Cd具有很强的耐性和积累的能力,可作为Cd污染土壤的修复植物.