太子参(PSEUDOSTELLARIA HETEROPHYLLA)病毒病病原鉴定

太子参(Pseudostellaria heterophylla)是我国传统的中药材之一,其病毒病十分严重,田间发病率和种根带毒率可达100％,平均90％左右。自太子参病株分离得到的T-17、T-32、T-69和T-01四个病毒分离物,经鉴定,它们分别为芜菁花叶病毒(Turnip mosaicvirus,TuMV)、黄瓜花叶病毒(Cucumber mosaic virus,CMV)、蚕豆萎蔫病毒(Broad beanwilt virus,BBWV)和烟草花叶病毒(Tobacco mosaic virus,TMV)。其中以TuMV分布最广,为害最重,是太子参病毒病的主要病原。

In Vitro Probiotic Effect of Pseudostellaria heterophylla Fibrous Roots on Feed Bacillus subtilis and Preliminary Study on Its Solid Fermentation

Traditional Chinese medicine Pseudostellaria heterophylla fibrous roots have been widely studied and applied in non-antibiotic clinical breeding of livestock and poultry. In vitro probiotic effect of P. heterophylla fibrous roots extract on feed probiotics Bacillus subtilis BD-K010 was studied, and the feasibility of solid fermentation of P. heterophylla fibrous roots was preliminarily evaluated. For in vitro probiotic effect, the increased concentration of P. heterophylla fibrous roots extract dependently increased the total biomass of B. subtilis BD-K010;1.0% P. heterophylla fibrous roots extract received the best effect, and the final p H of 1.0% experimental group was closer to neutral. Meantime, B. subtilis BD-K010 with optimum concentration of P. heterophylla fibrous roots showed significantly higher in vitro antibacterial effect than the control group(P<0.01),and the antibacterial effects on Escherichia coli and Staphylococcus aureus were improved by 51.99% and 63.16%, but it was ineffective to Salmonella. For solid fermentation, the profile of substrate complex and appendage flocculent structure on substrate surface at the end of fermenta-tion in experimental groups added with B. subtilis BD-K010 and cellulase plus BD-K010 were more complex;the live bacteria number, polysaccharide content and saponin content at fermentation end-point in two experimental groups were extremely higher than those in the control group(P<0.01).P. heterophylla fibrous roots extract had good in vitro probiotic effect on B. subtilis BD-K010 and promoted its antibacterial effect, and it is feasible to use probiotics for solid fermentation of P. heterophylla fibrous roots and to improve effective components. It is of great significance to further de-velop and utilize P. heterophylla fibrous roots resources in modern animal husbandry.

HETEROPHYLLIN-A AND B,TWO CYCLOPEPTIDES,FROM THE ROOTS OF PSEUDOSTELLARIA HETEROPHYLLA

Two cyclopeptides,heterophyllin A and B,have been isolated from the roots of Pseudostellaria heterophylla.Their structures were elucidated by chemical, spectroscopic,and enzymatic methods.

Reevaluation on Antioxidant Activity of Tonic Chinese Herbs Pseudostellaria heterophylla

Tonic Chinese herbs Pseudostellaria heterophylla has complex components,which has both medicinal and nutritional effects,and the clinical research on anti-oxidation of P.heterophylla has become a hotspot.By searching domestic and foreign databases,the antioxidant literatures of P.heterophylla were searched,consulted,sorted,classified and analyzed,and systematic reevaluation of antioxidant function was conducted from three aspects of P.heterophylla extract in vitro chemical method,biological method and animal test method according to evaluation criteria for antioxidant activity of traditional Chinese medicine and Meta analysis statistical methods.The results would provide systematic reference for further research and development of P.heterophylla,especially for forage value excavation and development of P.heterophylla in the field of modern animal breeding.

Investigation and Green Prevention and Control Strategies of Pseudostellaria heterophylla Diseases in Genuine Producing Area in Zherong County

Zherong County of Fujian Province is a genuine producing area of Pseudostellaria heterophylla,and disease has become a main factor restricting the healthy development of P.heterophylla industry in the county.In order to scientifically understand the occurrence status of agricultural diseases and serve the GAP of P.heterophylla,P.heterophylla diseases in Zherong County were investigated,and the information and regularity of diseases were summarized.P.heterophylla diseases in Zherong County were infectious,and caused by fungi and viruses.The diseases occurred frequently in spring and summer,and symptoms were manifested in pathological changes of leaf and tuber.Eventually,green prevention and control strategies of diseases were put forward:cultivating new disease-resistant varieties,scientifically administrating drugs according to symptoms,administrating drugs in optimum period,strengthening biological control,strictly requiring drug safety period and paying attention to green prevention and control during the planting process.The occurrence of P.heterophylla diseases in Zherong County was investigated and summarized,and the corresponding strategies were put forward,so as to scientifically serve the agricultural industry in P.heterophylla genuine producing area,and to provide a basis for the sustainable development of modern pollution-free planting industry of P.heterophylla.

Effects of Panax ginseng,Codonopsis pilosula and Pseudostellaria heterophylla on Sexual Maturity of SD Female Rats

[Objectives] To study the effects of Panax ginseng,Codonopsis pilosula and Pseudostellaria heterophylla on the growth and development of SD female rats. [Methods] The rats were randomly divided into control group,P. ginseng group,C. pilosula group,and P. heterophylla group,intragastrically administered with physiological saline,P. ginseng,C. pilosula,and P. heterophylla for 2 weeks,daily vaginal smear inspection was performed,estradiol( E_2) level before and after intervention was detected,rats were dissected,took out ovary and uterus,and calculated the ovarian index and uterus index. [Results]In 4 groups of rats,there was no significant difference in the time of sexual maturity and ovarian index; compared with the control group,E_2 level of C. pilosula group and P. ginseng group were higher( P < 0. 05);compared with the control group and P. heterophylla group,the ovarian index of P. ginseng group was higher( P < 0. 05); compared with the control group,the body weight of three administration groups increased much more( P < 0. 05). [Conclusions] P. ginseng and C. pilosula may lead to sexual maturity of SD female rats,while P. ginseng,C. pilosula and P. heterophylla can promote increase of body weight of SD female rats.

Key Points for High-yield Cultivation Management of Pseudostellaria heterophylla in Weihai Area

[Objectives] To explore the scientific cultivation mode of Pseudostellaria heterophylla in Shandong Province. [Methods] According to the ecological characteristics of P. heterophylla, the cultivation techniques and management points of P. heterophylla in Weihai are summarized. [Results] The scientific cultivation techniques and management points of P. heterophylla are introduced from the aspects of land selection, land preparation, seed selection, seedling nursing, timely sowing and transplanting, field management and disease and pest control. [Conclusions] This study will provide theoretical guidance for the high-quality, high-yield and high-efficiency cultivation of P. heterophylla in Shandong area.

Saponins Content in Fibrous Roots of Pseudostellaria heterophylla and Optimization of the SEP Extraction Technology

[Objectives]To explore the saponins resources in fibrous roots of Pseudostellaria heterophylla and optimize the SEP technology for extracting saponins.[Methods]Saponins contents in fibrous roots and root tubers of P.heterophylla were compared;with the fibrous roots as host materials,this experiment applied Box-Behnken design to analyze the influence of steam pressure,maintenance pressure time,moisture content of host materials on the content of saponins extracts.[Results]Saponins content was higher in fibrous roots than in tuberous roots of P.heterophylla;the response surface optimization showed that SEP(steam explosion pretreatment)promoted the contents of saponins and extracts in fibrous roots,and the optimal extracting conditions were stream pressure 1.50 Mpa,maintenance pressure time 54 s,and moisture content of host materials 30%.Under this extracting conditions,the verification value of saponins content experiment was 0.441%,36.53%higher than the untreated control group.[Conclusions]This research offered a new method of exploring saponins resources from fibrous roots of P.heterophylla,and provided a reference for developing new medicinal parts of P.heterophylla.

Pseudostellaria heterophylla:Antioxidant Activity and Its Mechanism of Action

The tonic traditional Chinese medicine Pseudostellaria heterophylla both medicinal and nutritional effects.It has been applied to research in the field of modern aquaculture,and its anti-oxidant clinical trials have also become a hot spot.Through searching domestic and foreign literature databases to sort out related literature,the anti-oxidation property of P.heterophylla was reviewed systematically from the three levels of in vitro chemical method,biological method and animal experiment method.On this basis,the functional mechanism of the anti-oxidant activity of P.heterophylla explored,in order to provide a reference for the follow-up in-depth research on the anti-oxidation of P.heterophylla and the discovery and development of feed and health care value of P.heterophylla the field of modern animal breeding.

太子参脱毒苗培养、化学成分及指纹图谱研究进展

太子参是我国的传统中药材,含有多种生物活性成分,如糖类、环肽类、皂苷类、微量元素、挥发油类等。现代药理学研究表明这些化学成分具有增强记忆、免疫调节、抑制食管癌、降血糖、促血管新生等活性。近年来,多地大力推广人工栽培,建设太子参GAP基地。但亦出现了一些问题,如太子参病毒病呈逐年加重、品种混乱、内在质量参差不齐等。针对这些问题学者们开展了诸多研究。本文从太子参脱毒苗研究、化学成分研究及指纹图谱研究等方面进行综述,以期为该资源的种植栽培、药物研发、质量标准建立等提供参考。

樟芝-太子参双向液体发酵培养基优化及效应分析

【目的】有效利用太子参根须副产物资源,提高樟芝(Antrodia camphorata)菌丝产量,促进活性成分合成。【方法】以太子参根须(粉末)作为培养基组分,Plackett-Burman试验筛选影响樟芝菌丝生长的关键成分,正交试验优化培养基组成。【结果】筛选试验结果表明,葡萄糖、酵母粉、蛋白胨及KH2PO4对樟芝菌丝生长有重要影响,正交试验优化的培养基组成为:太子参6 g·L^(−1),葡萄糖10 g·L^(−1),酵母粉4 g·L^(−1),蛋白胨6 g·L^(−1),KH_(2)PO_(4)1.5g·L^(−1)。利用优化培养基深层发酵,樟芝生物量达到4.73 g·L^(−1),与对照培养基及PDB培养基相比,胞内三萜含量分别提高2.75%和24.85%,胞外多糖分别提高161.11%和113.64%;发酵液中高密度无性孢子含量达1.8×10^(7)个·mL^(-1)。【结论】太子参适合作为樟芝双向液体发酵的药性成分,可有效促进樟芝菌丝生长、活性成分合成和无性孢子形成,是太子参根须副产物资源利用新途径。

太子参化学成分及生物活性研究进展

太子参为石竹科植物孩儿参的干燥块根,味甘、微苦,性平,归脾肺经,具有益气健脾、生津润肺之功效,常用于气虚津伤的肺虚燥咳,被国家卫生健康委员会批准用于保健食品。现代药理和临床研究表明,太子参中特征性化学成分多糖和环肽具有免疫调节、抗炎、改善记忆等作用。近年来对太子参的化学成分、结构和药理作用的综述仅局限于特征成分,对其他成分还鲜有系统梳理报道。因此本文系统梳理了太子参的化学成分及生物活性研究进展,以期为太子参的开发利用提供参考和依据。

植物乳杆菌发酵添加太子参的培养基优化及抗氧化活性研究

目的:优化植物乳杆菌(Lactiplantibacillus plantarum)发酵太子参的培养基,为太子参资源的利用和产品开发提供一定科学依据。方法:以菌体密度为指标,通过Plackett-Burman试验筛选对菌体细胞生长有重要影响的培养基组分,通过正交试验确定响应面试验的中心点,通过Box-Behnken响应面试验优化培养基组成;利用优化培养基发酵并测定发酵液(冻干粉)的抗氧化活性。结果:太子参水提物适合作为植物乳杆菌发酵的培养基成分;响应面法优化的培养基为麦芽糖29.7 g·L^(-1)、酵母粉27.5 g·L^(-1)、太子参水提物7.6 g·L^(-1)和MgSO40.5 g·L^(-1),优化条件下菌体密度达到1.11×10^(8)CFU·mL^(-1);优化培养基发酵液DPPH自由基清除率高于未发酵培养基(对照),低于MRS培养基发酵液(参照),ABTS+自由基清除率和FRAP值高于对照和参照。结论:太子参水提物可作为植物乳杆菌发酵的优良培养基成分,提高菌体密度;植物乳杆菌发酵可提高抗氧化活性。

太子参新品种‘贵参1号’生产适应性分析

目的:明确太子参新品种的生产适应性,为新品种的推广应用提供指导。方法:采用小区试验结合区域生产试验,在贵州省选取9个试验点对3个太子参品种的生长、抗病性、产量、品质进行考察。结果:与‘施太1号’和农家种相比,‘贵参1号’在株高、叶片大小、花朵数等地上部分农艺性状上有明显优势;病毒病病株率和病叶率显著降低,仅为2.53%和9.09%;药材平均亩产达到389 kg,多个试验点较农家种平均增产120%左右;药材形状近纺锤形,单个药材重量显著高于农家种,一、二等药材比例较‘施太1号’和农家种均显著增加,统货比例显著降低。药材质量均符合规定,水溶性浸出物含量45%、灰分2.6%均显著低于‘施太1号’;水分8.9%、环肽B含量0.0127%、多糖含量27%,均与‘施太1号’及农家种无显著差异。结论:综合考虑生长、抗病性、产量和品质,太子参新品种‘贵参1号’在贵州省东部到西部均有较强的适应性,建议作为优良品种在贵州省推广种植。

生物有机菌肥对太子参生长的影响

研究3种生物有机菌肥对太子参生长的影响,以期筛选出适宜太子参生长的生物有机菌肥种类。以当地传统施肥为对照,对比生物有机菌肥作为基肥使用对太子参生长的影响。结果表明生物有机菌肥作为基肥使用对太子参生长具有明显促进作用,无需再添加复合肥。适宜基肥组合为:复合微生物菌剂2400kg/hm(有效活菌数≥2.0亿)+豆粕微生物菌剂2400kg/hm^(2)(有效活菌数≥2.0亿)+定制套餐肥600kg/hm^(2)(有效活菌数≥1.0亿)+土壤调理菌剂600kg/hm^(2)(有效活菌数≥2.0亿)。生物有机菌肥作为太子参基肥使用增产增效显著,过量施肥反而不利于太子参生长。过量基肥组合为:复合微生物菌剂2400kg/hm^(2)(有效活菌数≥2.0亿)+豆粕微生物菌剂2400kg/hm^(2)(有效活菌数≥2.0亿)+定制套餐肥600kg/hm^(2)(有效活菌数≥1.0亿)+土壤调理菌剂600kg/hm^(2)(有效活菌数≥2.0亿)+复合肥750kg/hm^(2)。

太子参皂苷含量地理变异及气候响应特征

运用紫外-可见分光光度法测定4个产区药用植物太子参(Pseudostellaria heterophylla(Miq.)Pax ex Pax et Hoffm.)中的太子参皂苷含量,并分析太子参皂苷含量的地理变异规律,及其与气候因子在不同时间尺度上的响应关系。结果显示,太子参皂苷含量可分为江苏-安徽高含量区和贵州-福建低含量区。太子参皂苷含量与年均日照量呈极显著正相关,与年均相对湿度和年均降水呈负相关;与最热月份最高温呈显著正相关,与最冷季度平均温呈显著负相关。月均日照量和月均温对太子参皂苷含量影响较大,其中1-12月的月均日照量与太子参皂苷含量呈显著正相关;1-4月和11-12月的月均温与太子参皂苷含量呈负相关;5-9月的月均温与太子参皂苷含量呈正相关。产区间日照量差异可能是造成太子参皂苷含量地理变异的主要气候因素,日照量高、季度间温度差异大且相对少雨、低湿的地区有助于太子参皂苷的形成和积累。

不同施肥措施对太子参产量与品质的影响

【目的】探明不同施肥措施对太子参产量和品质的影响,为太子参的合理施肥提供科学依据。【方法】于2019—2021年通过田间定位试验,研究不同施肥措施〔施单层复合肥(T_(1))、施单层有机肥+复合肥(T_(2))、施双层复合肥(T_(3))、施双层有机肥+复合肥(T_(4))〕处理太子参的药材产量、生物总量、单株块根数、块根单重、块根长度及块根最大直径的变化,太子参药材优形指数和等级得率,折干率、水分、总灰分及浸出物的含量,进而分析双层施肥技术对太子参生长的影响。【结果】不同施肥措施对太子参产量和品质的影响不同,T_(2)和T_(4)对太子参生物总量(23.1 g/株、24.7 g/株)、折干率(37.7%、38.9%)和浸出物(37.1%、37.7%)影响显著,分别较T_(1)和T_(3)的生物总量显著提高17.9%、9.5%和26.0%、17.1%,折干率显著提高7.4%、6.5%和10.8%、9.9%,浸出物显著提高5.7%、5.1%和7.4%、6.8%;双层施肥的T_(3)和T_(4)对太子参产量(601.8 kg/667m^(2)、688.4 kg/667m^(2))、单株块根数(17.5块/株、18.1块/株)、块根长度(4.7 cm、4.6 cm)、块根最大直径(2.9 mm、3.2 mm)、优形指数(6.1、6.9)及一等品得率(11.2%、15.7%)影响显著,产量分别较T_(1)和T_(2)增产9.9%、0.8%和25.7%、15.3%,单株块根数增加18.2%、8.7%和22.3%、12.4%,块根长度降低9.6%、11.5%和7.8%、9.8%,块根最大直径增加26.1%、39.1%和16.0%、28.0%,优形指数增加38.6%、24.5%和56.8%、40.8%,一等品得率增加25.8%、76.4%和17.9%、65.3%。双层施肥较常规施肥显著降低太子参上层块根的长宽比、统货得率,增加上层块根一等品得率、折干率与浸出物,使上下层块根在形态及品质上更趋于一致。【结论】双层施肥有助于提高太子参产量及优品率,配施有机肥与复合肥则有助于提高太子参内在品质,采用双层施肥技术配施有机肥与复合肥适宜于太子参种植生产。

太子参多糖超声提取工艺优化

通过单因子试验和正交试验,研究了太子参多糖超声提取最佳工艺。结果表明:超声波法提取太子参多糖的最佳工艺是料液比1∶40(g/mL),提取温度70℃,超声功率500 W,浸提次数4次,提取时间30 min。优化后的多糖提取得率为6.77%,是优化前的3.7倍。优化后的提取工艺简单高效、成本低、稳定性好,可作为太子参多糖提取的理想方法。

Comparative chemical characters of Pseudostellaria heterophylla from geographical origins of China

Objective:Pseudostellaria heterophylla has been paid more attention in recent years,mainly as a medicine food homology plant.The content determination of P.heterophylla is not specified in the Chinese Pharmacopoeia(version 2020).The environmental conditions in different production areas could exert an influence on the quality of P.heterophylla.The purpose of this study is to discriminate P.heterophylla collected from different geographical origins of China.Methods:In this study,the content of polysaccharide in 28 batches of P.heterophylla was determined using phenol–sulfuric acid.HPLC fingerprints were established under optimised HPLC-PDA methods.Subsequently,the similarity analysis(SA)and the quantification of heterophyllin B were analyzed.The metabolites of P.heterophylla were identified and evaluated using UHPLC-Q Exactive HF orbitrap MS system.Principal component analysis(PCA),partial least squares discriminant analysis(PLS-DA),hierarchical cluster analysis(HCA)and orthogonal PLS-DA(OPLS-DA)were performed based on all peak areas.Results:The polysaccharide content in Guizhou and Jiangsu was higher than that of other production areas,which varied significant from different origins.While the content of heterophyllin B in Anhui and Jiangsu was high.The correlation coefficients of HPLC fingerprints for 28 batches samples ranged from 0.877 to 0.990,and the characteristic map can be used to identify and evaluate the quality of P.heterophylla.The samples from Fujian,Guizhou,Jiangsu provinces can be relatively separated using multivariate statistical analysis including PCA,PLS-DA,HCA,OPLS-DA,indicating that their metabolic compositions were significantly different.Ultimately,a total of 15 metabolites which were filtrated by a VIPvalue>1 and a P-value<0.05 associated with the separation of different origins were identified.Conclusion:HPLC fingerprint was established to evaluate the quality and authenticity of P.heterophylla.The present work showed that the difference of geographic distributions had an influence on the internal chemical compositions.A sensitive and rapid untargeted metabolomics approach by UHPLC-Q Exactive HF orbitrap MS was utilized to evaluate P.heterophylla from different origins in China for the first time.Overall,this study provides insights to metabolomics of P.heterophylla and supplies important reference values for the development of functional foods.

不同病毒侵染种根对太子参生长及产量和品质的影响

【目的】探明不同病毒对太子参生长、产量及品质的影响。【方法】以柘参2号为试验材料,采用RTPCR分子鉴定方法筛选出只携带一种常见病毒、携带两种常见病毒和不带病毒的太子参个体,经扩繁后,在田间使用随机区组的方法种植,探讨不同种类病毒对太子参的生长、产量和品质的影响。【结果】与未带病毒的植株相比,同时感染芜菁花叶病毒和蚕豆萎蔫病毒的植株总叶绿素含量、净光合速率和地上部株高分别降低了24%、33%和26%,单位面积的块根产量、浸出物含量和多糖含量分别降低了77.5%、76.2%和87.3%;而仅感染蚕豆萎蔫病毒的植株与未带病毒的植株相比,总叶绿素含量、净光合速率、地上部株高、块根产量、浸出物含量和多糖含量均无显著性差异。【结论】芜菁花叶病毒显著抑制太子参的生长,降低了太子参产量和品质,而蚕豆萎蔫病毒的影响不显著,在太子参脱病毒培养和脱毒苗的生产应用中应优先去除芜菁花叶病毒。