Effects of Different Initial Processing Methods on Atractylodin Content of Atractylodes chinensis

[Objectives]The paper was to compare the effects of different initial processing methods on atractylodin content of Atractylodes chinensis.[Methods]The atractylodin content of A.chinensis obtained by different initial processing methods was determined by HPLC.The loss rate on drying was determined by weighing.[Results]In the study of drying methods,the atractylodin content varied significantly among different thicknesses of slices,and the overall content of the product with the slice thickness of 5 mm was higher.Among different drying methods,constant temperature vacuum drying and shade drying of 5 mm slice resulted in the highest content of atractylodin.In the experiment of root impacting,root impacting twice received the best effect,and there was no significant difference in the atractylosin content of A.chinensis between drying in the sun and drying in the shade after root impacting twice(P>0.05).[Conclusions]Constant temperature vacuum drying or drying in the shade with the slice thickness of 5 mm,and root impacting twice is the best initial processing method,which leads to high atractylodin content of A.chinensis and good quality of medicinal materials.

Quality Evaluation of Atractylodes chinensis(DC.)Koidz.in Chengde Region Based on Dual Component Analysis

[Objectives]To evaluate the quality of Atractylodes chinensis(DC.)Koidz.in Chengde region.[Methods]35 samples of A.chinensis from different growth years were collected,and a high performance liquid chromatography(HPLC)method was established for the dual components of atractylodin and atractylone,and the quality evaluation was conducted on the cultivated A.chinensis from Chengde region in terms of moisture,ash and volatile oil content.[Results]The moisture content of all samples met the regulations,and the content of volatile oil was relatively high.Among them,seven samples with atractylodin content lower than 0.3%were unqualified products(accounting for 20%),and one sample had excessive ash content(accounting for 3%).[Conclusions]The established method for the determination of the dual components of Atractylodis Rhizoma is stable,simple,fast,precise and reusable.Besides,according to the Pearson correlation analysis among the indicators,it is concluded that the atractylodin and atractylone content are significantly related to the growth period and the volatile oil content.In addition,the quality evaluation indicators of A.chinensis in Chengde region show that there are differences in the quality of Atractylodis Rhizoma,but the overall pass rate is relatively high.It is necessary to further study the reasons for the differences.

Effect of Three Precursors on Accumulation of Principal Volatile Oil Constituents in Tissue Culture Plantlets of Atractylodes lancea (Thunb.) DC.

[Objective] This study was to explore the effect of three precursors on the accumulation of principal volatile oil constituents in tissue culture plantlets of Atractylodes lancea （Thunb.） DC. with the aim to provide references for the improvement of artificial cultivated A. lancea quality. [Method] Three precursors were added into the MS rooting medium for A. lancea tissue culture plantlets and the volatile oil was extracted by ultrasonication after cultured for several days. The content of atractylon, atractylol, β-eucalyptol and atractydin in the volatile oil were determined by using gas chromatography method. [Result] The addition of xylose, isoprene and tetrahydrofuran impacted the growth indicators, yield of volatile oil and relative percentage content of the four constituents of A. lancea tissue culture plantlets. In the 6 g/L xylose optimized medium, the atractylon and β-eucalyptol content reached up to 4.23% and 56.34%, respectively, 1.41% and 1.66% higher than the control; although the addition of isoprene into medium raised the atractylon content, the accumulation of total volatile oil was inhibited that it decreased by 23.67%, 31.06% and 7.10% to the control; for the tetrahydrofuran optimized medium, the content of atractylon, atractylol and atractydin all increased, and the total volatile oil content increased by 49.97% to the control when the concentration of tetrahydrofuran was 0.07 g/L. [Conclusion] The addition of xylose and tetrahydrofuran promoted the accumulation of principal constituents of the volatile oil, whereas the addition of isoprene inhibited the accumulation.

Effects of plug tray cell size on the growth of Atractylodes Chinensis (DC) Koidz seedlings

Atractylodes Chinensis(DC)Koidz is a perennial herb often used as a prescription medicine for influenza,to invigorate the spleen and remove dampness.The quality of the herb is determined by the quality of the seedlings.The traditional A.chinensis seedling production technique has nonuniform seedlings and low mechanisation.The plug seedling technique was used to produce A.Chinensis seedlings.This study was conducted to determine the growth characteristics of A.Chinensis seedlings according to the cell size of plug trays and number of days after sowing.Plant height,stem diameter,rhizome diameter,number of leaves,leaf area,and shoot dry and fresh weight of A.Chinensis seedlings were significantly higher in the P32-D treatment than in the P72,P50,and P32 treatments,but not significantly different from the P28-D treatment 60 d after sowing.The P28-D treatment resulted in a considerable drop in rhizome fresh weight and healthy seedling index,both of which are key indicators for assessing seedling quality.Although the differences in Pn among treatments were not significantly different,Tr was considerably greater in A.Chinensis seedlings treated with P32-D and P28-D than in the other three treatments.The P32-D and P28-D treatments had considerably greater potential maximum photochemical efficiency of PSII,quantum yield of photosystem II,photochemical quenching and electron transport rate than the other three treatments.Root vitality of A.Chinensis seedlings was significantly stronger in the P32-D treatment than in the other four treatments,and it was 1.9 times higher than in the P28-D treatment.The soluble protein,soluble sugar,and starch contents of A.Chinensis seedlings were highest in the P32-D treatment,but the differences among treatments were not significant.LUE,EUE,PY and EY were significantly higher in the P32-D treatment than in the other four treatments,being 1.3,1.3,1.4 and 1.4 times higher than in P28-D,respectively.On the other hand,as for the change in the growth of A.Chinensis seedlings according to the number of days after sowing,the growth of shoots and rhizome was most vigorous at the 60 d after sowing,while the fresh weight of the rhizome,which can be considered as an indicator of root growth,increased steadily during the experiment but slowed down after 60 d.As a result,32-cell deepened plug trays and a seedling nursery for a period of 60-75 d are recommended for commercial cultivation of A.Chinensis seedlings.This will provide technical support for production in A.Chinensis seedling.

Research Progress on the Efficacy and Chemical Components of Medicinal Taxillus chinensis(DC.)Danser

Taxillus chinensis(DC.)Danser is a traditional Chinese medicine,which has the effects of nourishing the liver and kidney and strengthening the bones and muscles.Its properties and efficacy were recorded in many ancient Chinese herbal medicine books and modern books.In addition,other parasitic plants of the Loranthaceae family also have the effects of promoting blood circulation,checking diarrhea,and resisting tumor to treat various diseases.This paper reviewed recent scholars’research on the efficacy and chemical composition of medicinal Loranthaceae plants,and expounded the similarities and differences between T.chinensis and other medicinal Loranthaceae plants.

北苍术对持续干旱的生理和转录组响应分析

为了探究北苍术响应干旱胁迫的生理和分子机制,本试验采用穴盘育苗法,分析持续自然干旱3~18 d北苍术响应干旱胁迫的生理特征;利用高通量转录组测序技术,分析干旱胁迫下北苍术的分子响应特征。结果表明,持续干旱6 d起,北苍术植株相对含水量极显著低于对照,可溶性糖、脯氨酸和丙二醛含量、抗氧化酶活性均显著或极显著高于对照;复水15 d时,北苍术过氧化氢酶(CAT)活性和植株相对含水量恢复到与对照无显著差异,但其他指标均仍极显著高于对照。主成分分析结果表明,可溶性糖和总蛋白含量、超氧化物歧化酶(SOD)和CAT活性可较好地反应北苍术的抗旱能力,干旱胁迫下北苍术隶属函数综合评价D值为0.765~0.865。转录组分析共获得14 538个差异表达基因(DEGs),GO功能注释最多的Unigene与细胞组分相关,KEGG分类中注释最多的Unigene是糖代谢;注释到萜类与聚酮类代谢途径的Unigene共71条,其中12条编码倍半萜和三萜类生物合成途径中的7个关键酶。综合分析可知,干旱胁迫影响北苍术相关功能基因表达,进而影响其生理特征。本试验为北苍术的抗旱机理和主要活性成分生物合成途径研究提供了参考。

不同叶面肥对茅苍术种子种苗生长及苍术素含量的影响

结合十堰市茅苍术[Atractylodes lancea(Thunb.)DC.]种子种苗生产需求,以一年生茅苍术种子种苗为材料,开展不同叶面肥对种苗的生物量和干燥根茎苍术素含量的影响研究。结果表明,不同叶面肥均未能促进一年生茅苍术种苗植株增高,对须根长和须根数无显著影响,减少了地下根茎鲜重和干重,相对提高了干重占比,但显著提高了一年生茅苍术种苗根茎的苍术素含量。此外,尽管叶面肥采用尿素能够提高一年生茅苍术种苗的叶片生长,但叶面肥采用磷酸二氢钾和硝酸钾有利于提高一年生茅苍术种苗的苍术素积累,提高了品质,更有利于大田生产应用。

北苍术与朝鲜苍术健脾作用及机制研究

目的 分析北苍术与朝鲜苍术干预后,脾虚大鼠血浆生物标志物及代谢通路的差异,从非靶向代谢组学的角度讨论两者的健脾机制。方法 将SD大鼠采用饮食不节、疲劳过度加苦寒泻下的复合因素法,造成脾虚症模型。测定胃肠道和免疫学指标的水平;采用超高效液相色谱串联Q Exactive质谱仪(UHPLC-QE-MS)技术,联合主成分分析(PCA)与正交偏最小二乘法判别分析(OPLS-DA)方法,寻找脾虚大鼠血浆中生物标志物;利用Pathway数据库归纳代谢通路。结果 经北苍术与朝鲜苍术给药后,脾虚大鼠血浆中各指标均有不同程度回调。代谢组学分析表明,北苍术与朝鲜苍术分别回调了70个和82个血浆差异代谢物,北苍术主要调节“甘氨酸、丝氨酸和苏氨酸代谢”与“硫胺素代谢”2个代谢通路,朝鲜苍术主要调节“甘氨酸、丝氨酸和苏氨酸代谢”、“硫胺素代谢”、“嘧啶代谢”、“丁酸代谢”与“核黄素代谢”5个代谢通路。结论 北苍术与朝鲜苍术对脾虚大鼠均具有一定的调节作用,其作用机制可能与调节“甘氨酸、丝氨酸和苏氨酸代谢”、“硫胺素代谢”等代谢通路有关。

北苍术叶片功能性状种源变异及其对科尔沁沙地环境的响应

【目的】探究北苍术叶片功能性状对科尔沁沙地的响应与适应规律。【方法】选择4个种源地北苍术半野生种子通过同质园实验育苗移栽于奈曼旗中(蒙)药材科研基地,以3年生北苍术叶片为材料,研究北苍术叶片结构与功能特性对科尔沁沙地的响应与适应规律。【结果】扎鲁特旗种源北苍术下部叶叶裂程度较明显,叶片革质化;北苍术叶上表皮细胞厚度、下表皮细胞厚度变异系数与可塑性指数均较大,围场县种源北苍术叶片上表皮细胞厚度、扎鲁特旗种源北苍术叶片下表皮细胞厚度显著高于其他种源,栅海比、组织结构紧密度较大;阿荣旗、扎鲁特旗种源地北苍术叶片上表皮气孔数量较多,喀喇沁旗种源上表皮气孔数量极少、气孔密度较小,喀喇沁旗种源叶片下表皮气孔数量显著高于其他3个种源地;围场县、扎鲁特旗种源地北苍术叶片光合能力较强。【结论】围场县、扎鲁特旗种源地的北苍术对科尔沁沙地有较强的适应能力。

苍术属植物分类研究进展

苍术属是菊科中重要的药用植物属,属内植物形态变异多样,种间和种下分类模糊不清。因此,明确苍术属系统分类、种间关系对属内药材资源开发利用具有重要作用。本文从苍术属系统分类、植物形态、细胞学、植物化学分类、分子系统等方面简要回顾苍术属分类研究成果,比较了不同学者对苍术属植物分类的分歧,对该属分类鉴定和种间关系进行了探讨和总结。苍术属植物形态的连续变化是植物形态分类分歧的根本原因,属内植物系统的分类要充分结合多学科或技术方法,并要考虑到资源利用的可持续性。本文结合属内药用植物的药材品质和临床用药,对苍术属植物分类研究提出了展望,以期为苍术属药用植物资源的保护利用提供参考。

北苍术固态发酵菌株的筛选、鉴定及发酵工艺优化

为筛选出适合北苍术固体发酵的菌株,以菌落数和粗多糖得率为筛选指标,获得适合北苍术发酵的乳酸菌,以含水量、接种量和发酵时间为因素,在单因素试验基础上,利用响应面法对发酵工艺进行优化。结果表明,从保存的菌株中共筛选出3株适合北苍术发酵的乳酸菌,通过生理生化试验以及16S rDNA测序确定该3株菌分别为乳酸片球菌(Pediococcus acidilactici)和戊糖片球菌(Pediococcus pentosaceus)。北苍术固态发酵的最佳条件为含水量50%、接种量5.5%、发酵时间2 d。在此条件下,北苍术粗多糖得率达到10.10%,相比未发酵组提高了37.45%。抗氧化试验表明,发酵能够显著提高北苍术粗多糖的DPPH自由基清除率和羟自由基清除率(P<0.05)。

基于形态性状和SSR标记的140份北苍术种质多样性分析

本研究以140份北苍术种质为材料,基于13个形态指标进行形态多样性分析,并利用SSR分子标记技术对其进行遗传多样性分析。结果表明,13个形态指标变异系数为1.93%~63.11%,其中主茎粗变异系数最大,花展开度的Shannon-Weiner多样性指数最大。主成分分析结果表明,种子长度、花展开度、主茎粗、株高、主茎与分枝夹角是反映北苍术形态多样性的主要指标。基于13个形态指标可以将140份北苍术种质聚为4个类群。利用筛选出的8对SSR引物扩增北苍术基因组DNA,根据特异性条带将140份北苍术种质聚为3个类群。13个形态指标和3个遗传多样性指标间相关性分析结果表明,单花结籽数与多态性信息量指数(PIC)呈显著正相关(P<0.05);主茎粗与Nei’s基因多样性指数(h)呈显著负相关(P<0.05)。综上,140份北苍术种质在形态指标和遗传多样性指标上均具有丰富的多样性,且形态指标与遗传多样性指标间具有一定的相关性。本研究为北苍术种质资源鉴定与利用提供了理论依据。

茅苍术根茎、须根挥发性成分比较

目的比较茅苍术根茎、须根挥发性成分。方法制作根茎、须根石蜡切片,光学显微镜下观察其横切面。水蒸气蒸馏法提取挥发油。GC-MS法检测苍术素、β-桉叶醇、茅术醇、苍术酮含量,并分析挥发性成分。采用多元统计分析软件进行主成分分析及差异代谢物筛选。结果根茎横切面可见大型油室,须根中可见油细胞。须根挥发油含量占根茎的24.88%,共检测出共有成分55种、根茎特有成分2种、须根特有成分6种。根茎中β-桉叶醇、茅术醇、苍术酮、苍术素含量比例为22.84∶1.16∶2.78∶1.00,须根中为5.59∶0.22∶0.97∶1.00。根茎、须根中挥发性成分具有显著性差异,组间区分度良好。根据OPLS-DA模型筛选出茅术醇、β-桉叶醇等12种差异代谢物。结论茅苍术根茎、须根挥发油成分类似,但含量不同,后者综合利用潜力更大。

北苍术种子萌发方法研究

[目的]研究提升北苍术种子萌发以及促进幼苗生长的方法。[方法]以北苍术为研究对象,对发芽率低的种子施用5种外源植物生长调节剂,以0 mg/L为对照,4种浓度(20、40、80、160 mg/L)处理种子24 h,测量种子萌发起始时间、发芽率、发芽势、种苗株高、主根长、侧根数量和种子简易活力指数。[结果]经过80 mg/L 6-BA、160 mg/L NAA、80 mg/L IBA、40 mg/L IAA、20 mg/L KT处理的北苍术种子的萌发能力及生长状况均得到显著提升,其中160 mg/L NAA处理效果最优。[结论]160 mg/L NAA处理种子发芽一致性、整齐性更好,适用于优质种苗繁育。

基于SCAR标记和DNA条形码技术的苍术基原鉴别研究

目的开发出能同时鉴别北苍术和关苍术的分子标记方法,并探究不同种质资源苍术的遗传进化关系。方法对不同地区北苍术Atractylodes chinensis(Bunge)Koidz及关苍术A.japonica Koidz.ex Kitam基因组DNA的差异片段进行测序,结合SRAP、ISSR、DAMD分子标记方法,优化PCR反应体系,筛选并转换成特异性标记,同时,采用条形码方法分析种间序列差异。结果通过SRAP、ISSR、DAMD三种分子标记方法的PCR扩增,共筛选出198对能稳定扩增且重现性好的引物,转换出7对能稳定、快速鉴别北苍术和关苍术的SCAR引物。条形码方法检测出北苍术ITS2序列长度为454 bp,关苍术ITS2序列长度为453 bp,与其他苍术属植物之间遗传距离较远。NJ树结果显示,北苍术、关苍术及其他苍术属植物均各自聚为一支,表现出良好的单系性。依据ITS2二级结构,4种苍术属植物在螺旋区的茎环数目、大小、位置均有明显差异,可以直观地进行区分。结论所开发的特异性SCAR标记为苍术属植物优良品种的筛选提供了新方法,DNA条形码能稳定、准确鉴别北苍术。

持续淹水对北苍术幼苗生理生化及抗氧化酶和活性成分生物合成相关基因表达的影响

为探究北苍术幼苗对持续淹水胁迫的生理生化和分子响应特征,以正常水分管理为对照,采用单因素完全随机试验设计,模拟持续淹水18 d和恢复正常水分管理后10 d处理条件下,北苍术幼苗在生理生化特性、根系活力及解剖结构、抗氧化酶活性及其相对表达和活性成分生物合成关键酶ACC基因的响应情况。结果表明:持续淹水2~18 d,北苍术幼苗水的相对质量分数在淹水第2,4,10,16,18天显著(P<0.05)高于对照。可溶性蛋白质量浓度、脯氨酸质量分数、丙二醛质量摩尔浓度随着淹水时间的增加呈现先升高后下降趋势,与对照呈显著差异;通气组织随着淹水时间的延长而增大;SOD,POD和CAT活性呈现先升高后下降趋势,其基因的相对表达量与对照差异显著,SOD和CAT相对表达量在淹水处理第4天达到最大值,POD相对表达量在淹水第10天达到最大值;活性成分生物合成关键酶基因ACC的相对表达水平与对照差异显著,在淹水第14天时达到最大值。恢复正常水分管理第10天时,生理生化指标、抗氧化酶活性和相关基因相对表达量均有所恢复,可溶性蛋白质量浓度、脯氨酸质量分数、丙二醛质量摩尔浓度仍与对照呈显著差异;通气组织明显缩小。北苍术幼苗通过调节渗透物质、抗氧化酶活性及相关基因表达和根系组织机构改变来适应淹水胁迫。

北苍术苗期除草剂的筛选

人工种植管理过程中杂草影响着北苍术苗期生长发育,寻找安全、高效、广谱的北苍术苗期除草剂。选用108 g/L高效氟吡甲禾灵乳油、250 g/L氟磺胺草醚、10%苯磺隆三种除草剂,通过外源喷施大田主要杂草(中华苦荬菜、狗尾草和马齿苋)和北苍术幼苗,筛选出适宜的除草剂浓度及合适配比,统计不同处理配比下对杂草和北苍术苗的防除效果。结果说明,108 g/L高效氟吡甲禾灵乳油对禾本科狗尾草防除效果好;250 g/L氟磺胺草醚对三种杂草防除效果均有效但对北苍术有损伤;10%苯磺隆对菊科中华苦荬菜防除效果好;采用0.023 mL/m^(2)108 g/L高效氟吡甲禾灵乳油+0.024 mL/m^(2)250 g/L氟磺胺草醚对北苍术伤害最小,对杂草防除效果最好。研究筛选出的除草剂可为北苍术的大田种植提供一定的理论指导与参考价值。

基于简单重复序列间标记分析寄主影响的广寄生遗传多样性

以寄生于不同寄主的广寄生[Taxillus chinensis(DC.)Danser]作为研究对象,运用简单重复序列间(Inter-simple sequence repeat,ISSR)分子标记技术对其进行遗传多样性分析。采用植物基因组DNA提取试剂盒对15个寄主的广寄生进行DNA提取,随后使用5个ISSR引物进行PCR扩增。对扩增出来的条带进行人工计数,借助POPGENE 3.2软件和NTSYS 2.10软件对15个寄主广寄生的遗传多样性进行分析。结果表明,共扩增出65条条带,均为多态性条带,多态性占比为100%。遗传相似系数在0.353 8~0.784 6,聚类图将15个样品分为6个类群,研究表明,15个寄主广寄生样品的遗传多样性存在差异,群体间具有遗传差异性。

Study on Rapid Propagation Medium of Atractylodes Lances by Orthogonal Design

[Objective] The aim was to protect wild resource of Atractylodes lances and to provide technical guidance for industrialization of Atractylodes lances. [Method] The test-tube seedlings were produced from seeds of Atractylodis Lances. At the same time, the effects of 6-BA, NAA and KT on the proliferation of aseptic seedling was studied through the orthogonal test. Effect of NAA concentration on the rooting of aseptic seedling was studied through nine types of light substrate groups, which consist of peat soil, fine sand and cotton seed hull, and their effects on Atractylodes lances growth were studied on with orthogonal design. [Result] The culture medium suitable for the proliferation of the Atractylodes lances tube seedling was MS＋6-BA 1.0 mg/L＋NAA 0.4 mg/L＋KT 0,4 mg/L. And the medium suitable for rooting was 1/2MS＋NAA 0.5 mg/L. The optimal ratio of soil, riversand and cotton seed hull was 3：3：2. [Conclusion] The research selected optimum culture medium, rooting medium and ratio of substrate, which provided technical guidance for laboratorial preservation of germplasm resource of Atractylodes lances and industrialization of its seedling production.

陇东旱塬区麦后柴胡荞麦套作栽培技术规程

为规范柴胡与荞麦套作种植关键环节的农事操作,促进产业的发展。通过多年试验示范,从范围、术语和定义、选地与整地、播种、田间管理、病虫害防治、收获及贮藏等方面总结制定了陇东旱塬区麦后柴胡荞麦套作栽培技术规程。