Notopterygium Incisum Extract Promotes Apoptosis by Preventing the Degradation of BIM in Colorectal Cancer

Objective Colorectal cancer(CRC),a prevalent malignancy worldwide,has prompted extensive research into anticancer drugs.Traditional Chinese medicinal materials offer promising avenues for cancer management due to their diverse pharmacological activities.This study investigated the effects of Notopterygium incisum,a traditional Chinese medicine named Qianghuo(QH),on CRC cells and the underlying mechanism.Methods The sulforhodamine B assay and colony formation assay were employed to assess the effect of QH extract on the proliferation of CRC cell lines HCT116 and Caco-2.Propidium iodide(PI)staining was utilized to detect cell cycle progression,and PE Annexin V staining to detect apoptosis.Western blotting was conducted to examine the levels of apoptotic proteins,including B-cell lymphoma 2-interacting mediator of cell death(BIM),B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(BAX)and cleaved caspase-3,as well as BIM stability after treatment with the protein synthesis inhibitor cycloheximide.The expression of BAX was suppressed using lentivirus-mediated shRNA to validate the involvement of the BIM/BAX axis in QH-induced apoptosis.The in vivo effects of QH extract on tumor growth were observed using a xenograft model.Lastly,APC^(Min+)mice were used to study the effects of QH extract on primary intestinal tumors.Results QH extract exhibited significant in vitro anti-CRC activities evidenced by the inhibition of cell proliferation,perturbation of cell cycle progression,and induction of apoptosis.Mechanistically,QH extract significantly increased the stability of BIM proteins,which undergo rapid degradation under unstressed conditions.Knockdown of BAX,the downstream effector of BIM,significantly rescued QH-induced apoptosis.Furthermore,the in vitro effect of QH extract was recapitulated in vivo.QH extract significantly inhibited the tumor growth of HCT116 xenografts in nude mice and decreased the number of intestinal polyps in the APC^(Min+)mice.Conclusion QH extract promotes the apoptosis of CRC cells by preventing the degradation of BIM.

GC-MS Analysis of Essential Oil Constituents from Rhizome and Root of Notopterygium forbesii

Aim To analyse the chemical constituents of the essential oil extracted from the rhizome and root of Notopterygium forbesii Boiss. and provide scientific basis for quality control. Methods The total essential oil was extracted by water-steam distillation and separated by capillary gas chromatography (GC). The components were determined by normalization method, and identified by GC-MS. Results GC-MS exhibited 217 peaks and 100 compounds were identified, accounting for 78.3% of the total essential oil. Conclu...

GC-MS Analysis of Essential Oil Constituents from Rhizome and Root of Notopterygium incisum

Aim To analyse the chemical constituents of the essential oil extracted from the rhizome and root of Notopterygium incisum Ting ex H.T. Chang, providing scientific basis for quality control. Methods The total essential oil was extracted by water-steam distillation and separated by capillary gas chromatography （GC）. The components were determined by normalization method, and identified by GC-MS. Results GC-MS exhibited 242 peaks and 83 compounds were identified, accounting for 75.77% of the total essential oil. Conclusion In the total essential oil of the rhizome and root of N. incisum, monoterpenes and sesquiterpenes accounted for 13.63% and 67.93%, respectively, in which （ 1S）-β-pinene （ 1.67% ）, 3-carene （ 1.05% ）, limonene （ 1.22% ）, and 1S-endo-bornyl acetate （ 1.68% ） as the monoterpenes and its derivatives, and （ ＋ ） -β-elemene （6.78%）, sativene （1.54%）, α-caryophyllene （2.64%）, germacrene D （1.67%）, eudesma-4 （ 14 ）, ll-diene （2.36%）, α-selinene （2.42%）, δ-cadinene （ 1.55% ）, 3,7,11-trimethyl-2,6,10-dodecatrien-1-ol （1.03%）, （ ＋ ）-elemol （5.18%）, （-）-spathulenol （1.40%）, guaiol （3.81%）, dehydroxy-isocalamendiol （ 1.06% ）, γ-eudesmol （ 1.05% ）, α-eudesmol （7.97%）, bulnesol （3.09%）, and carotol （2. 30% ） as the sesquiterpenes and its derivatives were main components. In addition, isopropyl transcinnamate was the maximum compound （ 11.3% ） of the total essential oil.

羌活(Notopterygium incisum)nrDNAITS和cpDNA rpl20-rps1序列分子进化特点的分析

应用PCR产物直接测序法分析了羌活居群间nrDNA(核糖体DNA)ITS序列和cpDNA(叶绿体DNA)rpl20-rps12的碱基差异,从而初步研究两套植物基因组的变异速率。采用改良的CTAB法从硅胶干燥的羌活叶片中提取总DNA,并对nrDNA ITS和cpDNA rpl20-rps12区域进行扩增、纯化、测序。nrDNAITS序列共有635 bp,有变异位点17处,变异位点百分率为2.68%,(G+C)含量为57.83%。cpDNA rpl20-rps12序列共有767 bp,有变异位点35处,变异位点百分率4.56%,(G+C)含量为33.06%。比较发现,羌活nrDNAITS区域较cpDNA rpl20-rps12序列保守,变异速率较慢。通过对ITS和rpl20-rps12序列单倍型(haplotype)进行分析发现,两者得出的结论一致,即现有分布范围经历了居群近期范围扩张。因此,羌活nrDNA ITS序列适合该种的谱系地理学研究。

Transcriptome analysis of the endangered Notopterygium incisum: Coldtolerance gene discovery and identification of EST-SSR and SNP markers

Notopterygium incisum C. C. Ting ex H. T. Chang(Apiaceae) is an endangered perennial herb in China. The lack of transcriptomic and genomic resources for N. incisum greatly hinders studies of its population genetics and conservation. In this study, we employed RNA-seq technology to characterize transcriptomes for the flowers, leaves, and stems of this endangered herb. A total of 56 million clean reads were assembled into 120,716 unigenes with an N50 length of 850 bp. Among these unigenes, 70,245(58.19%) were successfully annotated and 65,965(54.64%) were identified as coding sequences based on their similarities with sequences in public databases. We identified 21 unigenes that had significant relationships with cold tolerance in N. incisum according to gene ontology(GO) annotation analysis. In addition, 13,149 simple sequence repeats(SSRs) and 85,681 single nucleotide polymorphisms were detected as potential molecular genetic markers. Ninety-six primer pairs of SSRs were randomly selected to validate their amplification efficiency and polymorphism. Nineteen SSR loci exhibited polymorphism in three natural populations of N. incisum. These results provide valuable resources to facilitate future functional genomics and conservation genetics studies of N. incisum.

A new dimeric furanocoumarin from Notopterygium incisum

A new dimeric psoralen-type furanocoumarin, notopterol-（18-O-20＇）-notopol （1）, was isolated from Notopterygium incisum for the first time and its structure was elucidated by spectroscopic methods.

The chromosome-scale assembly of the Notopterygium incisum genome provides insight into the structural diversity of coumarins

Coumarins,derived from the phenylpropanoid pathway,represent one of the primary metabolites found in angiosperms.The alignment of the tetrahydropyran(THP)and tetrahydrofuran(THF)rings with the lactone structure results in the formation of at least four types of complex coumarins.However,the mechanisms underlying the structural diversity of coumarin remain poorly understood.Here,we report the chromosome-level genome assembly of Notopterygium incisum,spanning 1.64 Gb,with a contig N50 value of 22.7 Mb and 60,021 annotated protein-coding genes.Additionally,we identified the key enzymes responsible for shaping the structural diversity of coumarins,including two p-coumaroyl CoA 2′-hydroxylases crucial for simple coumarins basic skeleton architecture,two UbiA prenyltransferases responsible for angular or linear coumarins biosynthesis,and five CYP736 cyclases involved in THP and THF ring formation.Notably,two bifunctional enzymes capable of catalyzing both demethylsuberosin and osthenol were identified for the first time.Evolutionary analysis implies that tandem and ectopic duplications of the CYP736 subfamily,specifically arising in the Apiaceae,contributed to the structural diversity of coumarins in N.incisum.Conclusively,this study proposes a parallel evolution scenario for the complex coumarin biosynthetic pathway among different angiosperms and provides essential synthetic biology elements for the heterologous industrial production of coumarins.

基于聚类及PCA分析的羌活栽培区土壤质量评价

为了准确评价川产道地药材羌活栽培区耕作层土壤质量状况,分别采用聚类分析法(CA)和主成分分析法(PCA)构建栽培区耕作层土壤质量最小数据集(minimum data set,MDS),利用最小数据集土壤质量指数(soil quality index-CA,SQI-CA和SQI-PCA)和全量数据集土壤质量指数(SQI-T)评价川西北羌活栽培区耕作层土壤质量。结果表明:(1)羌活栽培区土壤有机质含量为(19.14±6.75)g·kg^(−1),处于中度贫瘠化水平;土壤速效氮、速效磷和速效钾含量较高,分别为(129.78±47.78)mg·kg^(−1)、(22.89±14.78)g·kg^(−1)和(159.87±97.87)mg·kg^(−1);土壤为中性土壤,pH均值为7.20±1.68。(2)基于不同数据集的土壤质量指数均值排序为SQI-T>SQI-PCA>SQI-CA,而SQI-PCA与SQI-T之间的Nash有效系数高于SQI-CA,相对偏差系数低于SQI-CA,说明基于主成分分析的最小数据集(MDS-PCA)评价效果更优,该数据集包括土壤容重、抗剪强度、有机质含量、饱和导水率、黏粒含量、pH、速效氮和砂粒含量共8个指标。(3)川西北羌活栽培区土壤质量指数SQI-PCA<0.33,表明该研究区耕作层土壤质量总体水平较差,主要体现在土壤紧实、有机质含量低,需要通过合理耕作、施肥和土壤改良等方式对耕作层土壤质量进行有效调控。研究结果可为川西北高原羌活栽培区土壤质量改良和生产适宜性调控提供参考,有利于川西北高原区中药材产区土壤可持续利用。

家种羌活主要化合物生物合成途径的转录组学研究

羌活是我国传统常用大宗药材,以根茎及根入药,前期研究发现其根茎中以羌活醇和异欧前胡素为代表的次生代谢产物含量显著高于根。为探究其生物合成途径及调控机制,本研究利用RNA-seq和转录组学分析手段,以根为对照,对比分析家种羌活根茎中基因的差异表达。结果表明,1222个差异表达基因(DEGs)显著富集于苯丙烷生物合成、植物病原体相互作用、植物MAPK信号通路及植物激素信号转导途径等通路;苯丙烷生物合成通路中莽草酸O-羟基肉桂酰转移酶(HCT)、肉桂醇脱氢酶(CAD)和过氧化物还原酶6(PRDX6)等关键酶编码基因转录本在根茎中显著上调表达;AP2/ERF、WRKY、C2C2及NAC等转录因子家族的多个转录因子在根茎及根中显著差异表达,这些因子可能是影响羌活次生代谢产物生物合成的关键转录因子;基因的差异表达与羌活根茎及根中羌活醇和异欧前胡素含量差异相符。本研究结果可为进一步研究羌活药用部位有效成分积累机理及定向育种提供理论基础。

Anti-rheumatoid arthritic effect of volatile components in notopterygium incisum in rats via anti-inflammatory and anti-angiogenic activities

Notopterygium incisum(QH) has been used for the treatment of rheumatoid arthritis(RA), and volatile oils may be its mainly bioactive constituents. The present study was designed to analyze the volatile compounds in QH and to determine the anti-arthritic capacity of Notopterygium volatile oils and the potential mechanism of action. The volatile compounds analysis was conducted by GC-MS. The anti-arthritic capacity test of the volatile oils was conducted on adjuvant-induced arthritis(AIA) rats. The anti-inflammatory property was tested in NO release model in RAW 264.7 cells. Endothelial cells were used to evaluate the anti-proliferative and anti-tube formative effects. 70 compounds were analyzed by GC-MS in the volatile oils. Notopterygium volatile oils weakened the rat AIA in a dose-dependent manner(2, 4, and 8 g crude drug/kg). The NO production by RAW 264.7 was decreased by more than 50% in Notopterygium volatile oils(5, 15, and 45 μg·mL^(-1)) pretreated groups. Notopterygium volatile oils also inhibited EAhy926 cell proliferation and further delayed EAhy926 cell capillary tube formation in a concentration-dependent manner. The anti-NO productive, anti-proliferative, and anti-tube formative effects of Notopterygium volatile oils strongly suggested that the therapeutic effect of QH in AIA might be related to the potent anti-inflammatory and anti-angiogenic capacities of the volatile oils.

HPLC法同时测定羌活药材中甲基托布津和辛硫磷农药的残留

目的建立同时测定羌活中甲基托布津和辛硫磷2种农药残留的HPLC法。方法羌活药材样品加NaCl粉末,以0.1%甲酸乙腈为提取溶剂,通过无水硫酸钠粉末净化,所得净化液以反相C_(18)色谱柱为分离柱,以乙腈∶0.1%甲酸(含10 mmol/L甲酸铵)(70∶30)为流动相,流速0.8 mL/min,在检测波长280 nm处测定2种农药残留的量。结果甲基托布津和辛硫磷配制浓度与峰面积在0.02~0.2μg/mL范围内有良好的线性关系,相关系数为0.9986和0.9993,该方法的检测限分别为4 ng/mL和3 ng/mL。甲基托布津的加标回收率为92.14%~112.56%,辛硫磷的加标回收率为87.85%~103.45%。结论该HPLC检测方法的操作简单、准确而快速,重复性和稳定性均良好,可满足羌活药材中甲基托布津和辛硫磷农药残留检测的要求。

Upper limb arthromyodynia can be treated with Notopterygium by down-regulating P2X3 to mediate PKC-induced inflammatory response

Rheumatoid arthritis(RA)is one of the most common refractory diseases in the world,and traditional Chinese medicine Notopterygium(NE)has been used in the treatment of upper limb pain for a long time.NE can significantly reduce the expression of inflammatory pain target P2X3 receptor in rats with upper-limb arthritis.To verify the relationship between the mechanism of NE for“upper limb paralysis”and the P2X3 receptor-mediated PKC inflammatory response pathway,UPLC was taken to measure the exact medicinal substance of ethyl acetate from NE.Sprague Dawley rats were randomly divided into a blank group,a model group,a live-action group,and a positive group.The joint cavity was removed after 21 d.Moreover,a model group,a live group,and a positive group were also set up with RA-FLS cells in our in vitro study.The expressions of P2X3 and PKC inflammation pathway indicators were detected by Western blotting analysis.A P2X3 inhibitor(A-317491)acted on RA-FLS cells,and a model group and a positive group were set.Then the protein expression of PKC was detected.NE reduced the expressions of P2X3,Rab7,PKC,and NF-κB at the protein level in both systems.NE and P2X3 receptor antagonists reduced the expressions of key proteins in the PKC pathway in RA-FLS cells to similar extents,and their effects were not additive.NE could effectively improve the“forelimb pain”of RA rats,with a mechanism closely related to the P2X3/Rab7/PKC/NF-κB pathway.

青藏高原野生宽叶羌活根际土壤微环境变化对光合作用的影响

为进一步研究青藏高原野生宽叶羌活药材形成对土壤环境的响应特性,明确土壤环境在宽叶羌活不同生长年限的差异,以不同生长年限野生宽叶羌活根际土壤为试验材料,研究根际土壤酶活性和有机碳含量的变化规律。结果表明,宽叶羌活对根围土壤孔隙度的影响主要在土层0~40cm深处,5年生地块平均土壤孔隙度减小至对照的88.56%,对土层50~100cm处影响较小。土壤含水量在土层0~50cm处变化显著,前3年根围土壤含水量逐年增大,达到对照的105.59%,土层60cm以下的土壤水分含量变化规律一致且差异不显著。土壤有机碳含量逐渐下降,从对照到5年生地块,土壤总有机碳含量降低到原来的41.75%,其中,可溶性有机碳含量降低到原来的51.78%,颗粒有机碳含量降低到原来的18.30%,活性有机碳含量降低到原来的24.42%,易氧化有机碳含量降低到原来的21.76%,微生物量碳含量降低到原来的47.31%;可溶性有机碳含量在3年生和5年生土壤间差异不显著,活性有机碳和易氧化有机碳含量在2年生和3年生土壤间差异不显著。随着生长年限增长,土壤脲酶、多酚氧化酶和过氧化氢酶活性呈先降低后升高的变化趋势,土壤脱氢酶和蔗糖酶活性没有明显变化趋势,土壤磷酸酶活性呈显著升高的变化趋势。随着株龄增大,叶片表观量子效率显著减小,光补偿点、光饱和点和暗呼吸速率均显著增大;叶片的原初光能转换效率、光化学淬灭系数和光合电子传递效率均表现不同程度的减小,非光化学淬灭系数表现显著增大,综合分析表明,随着生长年限的增加,土壤理化活性和部分土壤酶及有机碳含量降低,土壤微环境对根系生物活性的影响作用增大,根系生存环境的变化影响植株对土壤水分和营养物质的吸收,植株叶片同化效率降低,进而影响宽叶羌活药材质量和品质形成。

羌活种子不同休眠状态内生菌群落结构组成和差异性分析

为了探寻羌活种子不同休眠阶段内生菌的群落结构组成和差异性,利用Illumina Miseq平台分别对羌活种子不同休眠状态(形态生理双重休眠状态(GSA)、形态休眠解除状态(GSB)、生理休眠解除状态(GSC))细菌微生物16S V5+V7区域和真菌微生物ITS1-IF区域的扩增片段进行高通量测序,结合生物信息学进行分析。内生细菌的多样性较内生真菌丰富。内生菌群落种类在门水平上类似;在属水平上,从GSA到GSB阶段,内生菌群落组成发生变化,从GSB到GSC阶段,内生菌群落种类相似。通过Alpha多样性分析发现,从GSA到GSB再到GSC阶段,内生细菌群落多样性显著增大后直到种子生理休眠解除变化不变,而内生真菌群落多样性在种子整个休眠解除过程中变化不大。内生细菌群落结构从GSA到GSB再到GSC阶段均发生了显著性变化,而内生真菌群落结构从GSA到GSB发生显著变化后直到种子生理休眠解除,群落结构变化不大。LEfSe分析发现内生细菌共有37个差异丰富的分类学分支(a=0.01,LDA≥2.0),内生真菌则无。本研究明确了羌活种子不同休眠状态内生菌群落结构组成和差异性,丰富了羌活种子内生菌资源信息,对进一步研究利用微生物方法促进羌活种子休眠解除技术具有一定的指导意义。

羌活种子发育过程中形态及生理生化指标动态变化特征

【目的】探讨羌活种子发育过程中形态及生理生化指标变化规律,为羌活引种驯化、优质种质资源培育提供理论依据。【方法】采用定期观测采样及冷冻切片法,对不同发育阶段的羌活种子形态及生理生化指标进行测定。【结果】羌活种子发育过程中,种子大小、果翅厚度先升后降,果翅宽度升高,外种皮颜色由明亮的蓝绿色转为黄褐色;贮藏物质淀粉、可溶性糖、可溶性蛋白含量升高;淀粉酶活性小范围波动,琥珀酸脱氢酶活性升高,葡萄糖-6-磷酸脱氢酶活性交替升降,丙酮酸激酶活性呈“M”形变化,POD活性降低,而SOD活性先降后升;GA3、IAA和ZR含量降低,ABA含量升高;种子形态指标与SOD活性极显著负相关,贮藏物质含量与种子厚度极显著负相关,而与果翅宽度极显著正相关。【结论】在羌活种胚发育至心形胚前期过程中,外种皮由蓝绿色转为黄褐色,贮藏物质含量升高,关键酶活性波动变化,促生长激素含量波动降低,而胁迫激素含量升高。

Cytotoxic polyacetylenes isolated from the roots and rhizomes of Notopterygium incisum

Phytochemical investigation on the roots and rhizomes of Notopterygium incisum led to the isolation of a new polyacetylene, notopolyenol A (1), along with thirteen known analogues (2-14). Their structures were elucidated by extensive analyses of NMR and HRMS data, and the absolute configuration of 1 was unambiguously determined as 3 R by comparison of its retention time and ECD curve with those of synthetic enantiomers (-)-1 and (+)-1, whose absolute configurations were established by using the modified Mosher's method. Subsequent activity screening revealed that (3 S)-1 exhibited the most significant cytotoxicity against MCF-7, H1299, and HepG2 cancer cells with IC_(50) values of 1.3 μmol/L,0.6μmol/L and 1.4μmol/L, respectively.

羌活根际土壤微生物群落结构特征及其对次生代谢产物的影响

[目的]科学评估羌活主产区土壤生态系统健康状况,揭示土壤微生物与羌活的互作关系。[方法]以羌活根际土壤为研究对象,分别采用高通量测序技术和高效液相色谱法(HPLC)测定土壤微生物群落结构组成和羌活次生代谢产物含量,进一步采用冗余分析和相关性分析方法研究根际土壤微生物对羌活次生代谢产物的影响。[结果]羌活根际土壤细菌优势门类群为变形菌门(Proteobacteria)、酸杆菌门(Acidobacteria)和拟杆菌门(Bacteroidetes),真菌优势门类群为子囊菌门(Ascomycota)、担子菌门(Basidiomycota)和球囊菌门(Glomeromycota)。属水平上,细菌优势属为Vicinamibacter(0.80%~9.11%)、土生单胞菌属Terrimonas(0.66%~7.72%)和硝化螺旋菌属Nitrospira(1.18%~4.46%);真菌优势属为被孢霉属Mortierella(0.01%~14.19%)、四孢菌属Tetracladium(0.18%~14.32%)和黄豆菌属Kotlabaca(0.01%~24.96%)。冗余分析表明羌活根际土壤微生物群落结构主要受土壤pH及养分含量影响,其中Colletotrichum和Botryichum与pH呈显著负相关(P<0.05),Bradyrhizobium与有机质亦呈显著负相关(P<0.05),Candidatus_Udaeobacter、Scutellospora和Enterocarpus均与速效磷和速效氮呈显著正相关(P<0.05)。羌活次生代谢产物羌活醇与毛壳菌属(Chaetomium)、丝盖伞属(Inocybe)和米拉珊瑚属(Melanophyllum)呈显著负相关(P<0.05),异欧前胡素则与毛壳菌属(Chaetomium)和腐质霉属(Humicola)呈显著正相关(P<0.05)。[结论]该研究结果为科学评估羌活主产区土壤生态系统健康状况和微生物-羌活互作关系提供数据支撑。

层积过程中羌活种子种胚形态及生理生化变化

对羌活(Notopterygium incisum Ting ex H.T.Chang)种子进行层积处理,观测层积过程中种子种胚形态及胚率,研究种子中营养物质和内源激素含量以及呼吸途径关键酶活性的变化;并对层积后的种子进行氟啶酮处理,测定种子的发芽率和发芽势。结果显示:与未层积(层积0 d)相比,层积后羌活种子种胚由球形发育为子叶型,胚率增长至67.30%,发芽率增长至50.67%;种子的蛋白质、可溶性糖和淀粉含量显著降低;赤霉素(GA_(3))和吲哚乙酸(IAA)含量分别在59.15~81.06和14.06~22.08 ng·g-1范围内波动变化,脱落酸(ABA)含量在层积150和180 d时显著降低,GA_(3)/ABA比和(GA_(3)+IAA)/ABA比在层积120~180 d显著升高。在层积0~60 d,羌活种子的丙酮酸激酶(PK)活性呈先升高后降低的趋势,琥珀酸脱氢酶(SDH)活性明显升高,葡萄糖-6-磷酸脱氢酶(G-6-PDH)活性先降低后略有升高,该时期以三羧酸循环途径(TCA)为主;在层积60~150 d,PK活性呈波动升高的趋势,SDH和G-6-PDH活性总体呈波动降低的趋势,该期间以糖酵解途径(EMP)为主;在层积180 d时,PK、SDH和G-6-PDH活性均显著高于层积120 d。相关性分析结果显示:ABA含量分别与胚率和发芽率呈显著和极显著负相关,相关系数分别为-0.800和-0.875。综合研究结果表明:羌活种子具有胚后熟特性,层积处理结合氟啶酮处理可打破羌活种子休眠,提高种子的发芽率;营养物质含量的降低,呼吸途径关键酶活性的波动变化,ABA含量的降低以及GA_(3)/ABA比和(GA_(3)+IAA)/ABA比的升高对羌活种子休眠解除有重要影响。

四川栽培羌活表型性状与品质相关性分析

为了探究川产高海拔地区栽培羌活(Notopterygium incisum Ting ex H.T.Chang)的植物表型性状与品质性状(羌活醇、异欧前胡素)的相关性,提高良种选育效率,本研究选取3个羌活栽培基地的四年生羌活植株183份,统计13个表型性状,并测定地下部分的羌活醇与异欧前胡素含量,通过聚类分析、冗余分析和方差分析法解析表型性状与品质性状的内在关联,筛选高品质羌活的定向选育表型性状。结果表明,基生叶片数变异系数和多样性指数在所有调查性状中最高,基生叶片表型性状以绿色、纸质、深裂、锯齿状为主,基生叶柄和花茎表型性状表现为绿色、紫色、绿色带紫纹均有分布,基生叶柄绿色和花茎浅紫色最多。聚类分析和冗余分析结果表明,地下部分羌活醇和异欧前胡素含量与基生叶片及基生叶柄颜色显著相关(P<0.05)。方差分析结果表明,以基生叶片深绿色、叶柄紫色为主要特征的羌活植株地下部分羌活醇、异欧前胡素含量及其总含量均最高。综上,羌活植株基生叶片颜色和叶柄颜色可作为羌活药材指标成分羌活醇及异欧前胡素高含量品种的定向选育表型性状。本研究结果为解决人工栽培羌活药源危机提供了有效途径,为优良种质的定向选育提供了参考依据。

当归和羌活根际促生菌筛选及特性研究

为获取对当归和羌活生长具有促生作用的促生菌(plant growth promoting rhizobacteria,PGPR),以当归、羌活的根系为研究对象,利用选择性培养基分离具有固氮、溶磷和分泌3-吲哚乙酸(IAA)能力的菌株。通过对菌株的固氮酶活性、溶磷能力及分泌IAA的能力进行测定,筛选出促生性能优良的菌株;进一步通过生理生化指标和16S rDNA序列分析相结合的方法分析优良菌株的分类地位。结果表明,共筛选到12株具有固氮酶活性的菌株,其固氮酶活性为0.30~4.04 nmol C2H4·mL^(-1)·h^(-1);6株溶解无机磷的菌株,溶磷量为290.98~420.33μg·mL^(-1);3株产IAA的菌株,IAA分泌量为10.38~18.63μg·mL^(-1)。经鉴定,溶磷特性优良的菌株属于假单胞菌属(Pseudomonas)和错玫杆菌属(Falsirhodobacter),假单胞菌属为优势菌属。综合各菌株的促生特性,筛选出2株促生性能优良的菌株:WQP-5(Pseudomonas grimontii)和MDP-8(Pseudomonas thivervalensis),为开发适用于甘肃道地药材当归与羌活专用生物菌肥提供了优良菌种。