Structural Characterization of Chloroplast Genome in Alpinia japonica(Thunb.)Miq.,a Medicinal Plant of the Genus Alpinia

The analysis of chloroplast gene characteristics in Alpinia japonica(Thunb.)Miq.is of great significance for developing relevant genetic resources.The high-throughput sequencing and bioinformatic research were performed to analyze the chloroplast genome characteristics of A.japonica.The total chloroplast genome length of A.japonica was 161,906 bp,with a typical circular tetrameric structure.And 133 genes were annotated,comprising 87 protein-coding,38 tRNA,and 8 rRNA genes.Furthermore,22 genes contained two copies,and 18 genes owned introns.Repeat sequence analysis showed that it contains 321 simple sequence repeats(SSRs)and 37 long segment repeats.Compared with the chloroplast genomes of eight representative plants in the genus Alpinia,the gene structure,type,and quantity were relatively conservative.Rps12 was the highest variation site in the entire chloroplast gene.A phylogenetic tree showed that the genus Alpinia was the most closely related to the genus Amomum.Meanwhile,A.japonica is the most closely related to Alpinia chinensis belonging to the genus Alpinia.Overall,the chloroplast genome of a new species was reported in the genus Alpinia,and a basis was provided for the utilization of Alpinia plants as a medical resource.

Effects of Different Seed Stem Sizes on the Changes of Available Elements in Rhizosphere Soil of Fritillaria thunbergii Miq.

[Objectives]This study was conducted to screen suitable seed stems of Fritillaria thunbergii Miq.from different provenances and to provide a theoretical basis for the high-yielding and high-efficiency cultivation of F.thunbergii Miq.introduced to different places.[Methods]F.thunbergii Miq.from four different provenances including Zhejiang,Nantong and Chongqing were introduced and cultivated in Wanzhou of Chongqing.The contents of available Zn,Fe,Mn,Cu,Mo,N,P,K,Ca and Mg in rhizosphere soil of F.thunbergii Miq.during five growing stages were determined after selecting different stem sizes for field cultivation.[Results]Small stems of Pan an and Ningbo provenances(SSG3,121-160/kg)and middle stems of Nantong and Fengjie provenances(SSG2,81-120/kg)showed higher soil availability.[Conclusions]In the process of introduction and cultivation of F.thunbergii Miq.,high yield and high efficiency can be achieved by selecting smaller seed stems of F.thunbergii Miq.

Constituents from Ranunculus sieboldii Miq.

Aim To investigate the chemical composition of Ranunculus sieboldii Miq..Methods Repeated column chromatography over silica gel, polyamide and RP-18 followed by gelfiltration on sephadex LH-20 were used to isolate chemical constituents, and their structures wereelucidated by extensive spectroscopic methods (UV, IR, MS, ~1H NMR, ^(13)C NMR) including 2D NMR(COSY, HMQC, HMBC, NOESY) techniques and by direct comparing spectral data with those reported inliterature. Results Five flavonoid glycosides named apigenin-4'-O-α-L-rhamnopyranoside (1),apigenin-7-O-β-D-glucopyranosyl-4'-O-α-L-rhamnopyranoside (2), apigenin-8-C-α-L-arabinopyranoside(3), apigenin-8-C-β-D-ga-lactopyranoside (4) , tricin-7-O-β-D-glucopyranoside (5), together withtricin (6), luteolin (7), scopoletin (8), esculetin (9), scoparone (10), ferulic acid (11),protocatechuic acid (12) , and tematolide (13) were isolated from the 95% etha-nolic extract of itswhole plant, and their cytotoxic activities were preliminarily tested. Conclusion Compounds 1-12were obtained from this genus and compound 13 from this species for the first time. Furthermore,compound 1 was for the first time isolated from nature while the ^(13)C NMR data of compounds 2 and3 are reported for the first time. The bioassay revealed that compound 1 was active against BEL-7407and A549 cell lines (IC_(45) 43, 77 μg·mL^(-1)), 8 and 10 showed inhibitory activities on KB celllines (IC_(50) 78, 44 μg·mL^(-1)) and HL-60 cell lines (Ic_(50) 85, 85 μg·mL^(-1)), while 7exerted moderate cytotoxic activities on KB, BFL-7407, A549 and HL-60 cell lines with their IC_(50)being 51, 55, 44 and 10 μg·mL^(-1) , respectively.

Study on Germination Rate of Uncaria rhynchophylla (Miq.) Jacks

[Object] The research aimed to find out seed germination conditions and regulations.[Method] Seeds were soaked by clear water and 4 chemical agents of different concentrations(NaHCO3,KHCO3,K2CO3,NaClO)then conducted germination test in illumination box.[Result] Seed germination number and absolute germination rate(above 80%)in water treatment group was obviously higher than other treatments.[Conclusion] No dormancy phenomenon in mature embryo and no inhibition in seed coat,so conventional moist treatment can induct seed germination.

竹叶眼子菜(POTAMOGETON MALAIANUS MIQ.)叶面积测定及与栽培密度的关系初探

通过人为控制种群密度的栽培实验 ,用叶特征度量估测单叶叶面积 ( AL)、用茎特征度量估测整株叶面积 ( AS) ,分析不同栽培密度对叶面积测定的影响。结果表明 :( 1 )叶长×叶宽 ( L× W)与单叶叶面积相关性最大 ;茎长×茎上叶片数 ( LS× NL)测定整株叶面积较为适宜。 ( 2 )随着栽培密度增大 ,叶片相对变长变窄 ,比叶面积则增加 ,平均整株叶面积相应减小。( 3)不同的栽培密度对叶片形状有明显影响 ,反映在不同密度处理条件下叶片的几何拟合图形不同。

Comparison between Sulfuric Acid-phenol and Sulfuric Acid-anthrone Methods Used for Determination of Polysaccharides in Shoots of Aralia elata (Miq.) Seem.

Sulfuric acid-phenol and sulfuric acid-anthrone methods were used to detect polysaccharide content in shoots of Aralia elata（ Miq.） Seem.,and the conversion factor to glucose was measured with refined polysaccharides. Comprehensive evaluation was carried out by linear relationship,precision,reproducibility,stability and recovery rate. The results showed that the linear relationship between glucose concentration and absorbance was good when glucose concentration was0-40 μg/ml,and the average recovery rate was equal to or higher than 97. 00% with good reproducibility（ RSD 〈 1. 60%,n = 5）. It revealed that the two methods were accurate and reliable,and suitable for the determination of polysaccharide content in the shoots of A. elata. Polysaccharide content detected by sulfuric acid-phenol and sulfuric acid-anthrone methods was 19. 31% and 20. 40% respectively.

Current Situation of Fritillaria thunbergii Miq. Production in Yinzhou District of Ningbo City

Yinzhou District of Ningbo City is the home and main production area of Fritillaria thunbergii Miq. Based on the analysis of Fritillaria thunbergii Miq. production in Yinzhou District in 2015,this paper firstly introduced current situation of Fritillaria thunbergii Miq. planting. The competition of Fritillaria thunbergii Miq. products was in disorder,the market price in 2015 dropped to half in 2014,and many Fritillaria thunbergii Miq. growers suffered losses. To keep sustainable development of traditional Fritillaria thunbergii Miq. industry in old production areas and maintain brand and name of genuine materials of " Zhejiang 8 Famous Kinds Herbal Medicines",it is required to actively reduce the inventory,make up short board,strengthen advantages,implement the reform of supply font,keep up with the times and market,and succeed through improving the quality.

Study on Endophytic Bacteria Diversity in the Root of Peperomia dindygulensis Miq. by Applying Clone Library Construction Method

[Objectives] To study the endophytic bacteria diversity in the root of Peperomia dindygulensis Miq. [Methods] The root of P.dindygulensis Miq. was taken as materials,culture-independent method was applied to build 16 S rDNA clone library for endophytic bacteria of P. dindygulensis Miq. [Results] In the 16 S rDNA clone library for endophytic bacteria of P. dindygulensis Miq.,95 clones separately belonged to 46 OTUs,Phylogenetic analysis indicated that these cloning sequences separately belonged to phylum Proteobacteria,phylum Bacteroidetes,phylum Actinobacteria,phylum Nitrospirae,and phylum Gemmatimonadetes. In these,phylum Proteobacteria accounted for 70. 53% of total number of clones,so it was the most dominant group in this library. [Conclusions]The endophytic bacteria diversity in the root of P.dindygulensis Miq. is rich and this study is expected to provide scientific basis for protection,development and use of resources of P. dindygulensis Miq.

Effect of extracts from Stachys sieboldii Miq. on cellular reactive oxygen species and glutathione production and genomic DNA oxidation

Objective: To evaluate the antioxidant activity of extracts and fractions from Stachys sieboldii Miq., and to examine its effect on the cellular reactive oxygen species(ROS) and glutathione(GSH) production and genomic DNA oxidation in HT-1080 cells. Methods: The ROS generation induced by H2 O2 was measured by the dichlorofluorescein-diacetate assay. GSH levels were measured using a fluorescent method with mBBr. Genomic DNA oxidative damage was measured with levels of oxidative DNA induced by the reaction of ferritin with H2 O2. Results: The n-hexane, 85% aqueous methanol and n-butanol fractions(0.05 mg/mL concentrations) inhibited H2 O2-induced ROS generation by 63%, 35% and 45%, respectively. GSH levels were significantly increased in both acetone+methylene chloride and methanol extracts(P<0.05). Supplementation of cells with n-hexane significantly increased GSH levels at concentrations of 0.05 mg/mL(P<0.05). Both the acetone+methylene chloride and methanol extracts, as well as all fractions significantly inhibited oxidative DNA damage(P<0.05). Conclusions: These results indicate that cellular oxidation was inhibited by the n-hexane fraction and this fraction may contain valuable active compounds.

A New Flavanone from the Bark of Morus macoura Miq.

A new flavanone named macrouroine C (1) was isolated from the bark of Morus macroura Miq.. The structure of 1 was elucidated mainly on the basis of spectroscopic evidence.

Total Saponins of Aralia elata （Miq.） Seem Leaves （TSAESL） Inhibits Tumor Cell Proliferation and Induces Apoptosis via activating MAPK signaling pathways in MCF-7 human breast cancer cells

Aim Aralia elata （Miq.） Seem is a medicinal plant with a variety of biological activities. Several trier- pene saponins extracted from the leaves of Aralia elata （Miq.） Seem possess potent cytotoxic activity, but the mechanisms remain unclear. Methods In the present study, we examined the anti-tumor activity of total saponins of Aralia elata （Miq.） Seem leaves （TSAESL） using both in vitro and in vivo experiments, and investigated the underlying molecular mechanisms. Results Briefly, TSAESL significantly inhibited the proliferation of five human cancer cell lines and induced apoptosis of MCF-7 cells in a concentration-dependent manner. TSAESL also in- creased the expression level of active caspase-3 and cleaved PARP. In addition, TSAESL dramatically activated the phosphorylation of ERK1/2, p38 and JNK in a concentration-dependent manner. Treatment with the ERK1/2 in- hibitor U0126, p38 inhibitor SB203580, or JNK inhibitor SP600125 prior to TSAESL exposure markedly attenuated TSAESL-induced phosphorylation of ERK1/2 and p38. In mice bearing MCF-7 xenografl, TSAESL markedly inhib- ited tumor growth without significantly affecting spleen coefficient and hematological parameters. Conclusion These results strongly suggest that TSAESL has anti-tumor effect via activating the MAPK-mediated signaling path- ways.

Effects of Different Harvest Periods on Seed Quality of Alpinia oxyphylla Miq.

[Objectives] To clarify the appropriate harvest period of Alpinia oxyphylla Miq. seed and improve the quality of seeds. [Methods] In this experiment,the basic parameters and germination characteristics of A. oxyphylla Miq. seeds during different harvest periods were analyzed and compared. [Results] The results showed that the A. oxyphylla Miq. fruit picking period was closely related to the seed quality,and the seed maturity was highest 100 d after the flowering of A. oxyphylla Miq.,that is,the A. oxyphylla Miq. fruit skin turned from green to yellow( yellow-green),the peel had a small number of brown spots,it was spicy enough,the fruit dry/fresh weight ratio was greater than0. 34,the seed thousand kernel weight reached 11 g or more,and the dried seed moisture content was 13. 01%. Using the TTC method,the seed viability index was determined to be 85%,and in this case,it was the best time of harvesting seed. The water permeability of A. oxyphylla Miq. seed was poor,and the seed imbibition and water absorption time was long,about 72 h. The maximum water absorption rate was about23. 71% at the imbibition and water absorption stage,and it was inferred that the A. oxyphylla Miq. seeds were hard seeds. The germination time of A. oxyphylla Miq. seeds was long,the mature A. oxyphylla Miq. seeds began to germinate on the 12 th d,and the germination rate reached 75. 56% on the 50 th d. [Conclusions] This study determined the optimum harvest period of A. oxyphylla Miq. seeds,clarified the biological characteristics of slow seed water absorption and long germination cycle,and concluded that the seed coat was the main factor leading to slow seed water absorption and long seed germination cycle.

Temperature Effects on Seed Dormancy Relieving of Fritillaria thunbergii Miq

The seed dormancy relieving of Fritillaria thunbergii Miq required 50 days at 8～10℃and 80 days at 3～5℃ In order to study the physiological basis for seed dormancy relieving,peroxidase,acid phosphatase and proteins were assayed by spectrophotometric analysis and electrophoresis The results showed that peroxidase activity and acid phosphatase activity increased and new bands were detected in protein pattems as the seeds were treated with low temperature,these changes resulted in the seed dormancy relieving

Optimization of Extraction Process of Polysaccharides from Actinidia arguta(Sieb.&Zucc.)Planch.ex Miq.

[Objectives]To optimize the extraction process of polysaccharides from Actinidia arguta(Sieb.&Zucc.)Planch.ex Miq.[Methods]On the basis of single-factor tests,an orthogonal test was conducted with solid/liquid ratio,ultrasonic temperature,ultrasonic time and ultrasonic power as influencing factors.[Results]The optimal extraction process was as follows:solid/liquid ratio 1∶15(g/mL),ultrasonic temperature 50℃,ultrasonic time 50 min and ultrasonic power 90 W.Under the optimal extraction conditions,the extraction rate of polysaccharides reached 2.05%.[Conclusions]The method is simple and feasible,and can be used to optimize the extraction process of polysaccharides in A.arguta,in order to provide a theoretical basis for the further development and utilization of related resources.

山银柴胡质量标准研究

目的:研究山银柴胡质量评价标准,建立指纹图谱分析方法。方法:参照2020年版中国药典的方法测定山银柴胡水分、总灰分、酸不溶性灰分、醇溶性浸出物含量;采用HPLC法测定山银柴胡中成分含量;建立30批山银柴胡的指纹图谱,结合相似度评价、聚类分析、正交偏最小二乘判别分析(OPLS-DA)进行质量评价。结果:30批山银柴胡的水分、总灰分、酸不溶性灰分、醇溶性浸出物、丝石竹皂苷元3-O-β-D-葡萄糖醛酸甲酯、丝石竹皂苷元、皂皮酸含量的平均值分别为8.89%、12.67%、0.79%、34.53%、13.58 mg/g、1.62 mg/g、1.03 mg/g;指纹图谱确定了17个共有峰,样品与对照图谱的相似度为0.916~0.997,聚类分析可将样品分为3类,OPLS-DA分析筛选出7个质量差异性成分。结论:该研究建立的方法可为山银柴胡质量标准的完善提供参考。

石榕树中补骨脂素分离鉴定及其抗真菌活性

[目的]分离出石榕树中能够有效抑制植物病原真菌的活性单体化合物并探索其抗真菌活性。[方法]采用生物活性追踪和化学分离相结合的方法对石榕树甲醇提取物进行分离,运用质谱、核磁共振氢谱和碳谱并结合相关文献数据鉴定活性单体化合物。采用菌丝生长速率法测定活性单体化合物对7种植物病原真菌的抑菌活性。[结果]石榕树(叶)甲醇提取物对甘蔗凤梨病菌具有良好抑菌活性,在10 mg·mL^(-1)浓度下,抑菌率为100%。在浓度为1.5 mg·mL^(-1)时,石榕树(叶)石油醚、乙酸乙酯、正丁醇萃取物和水层萃余物对甘蔗凤梨病菌的抑菌率分别为58.73%、100.00%、39.49%和6.63%。表明石榕树(叶)乙酸乙酯层萃取物对甘蔗凤梨病菌的抑制作用最高,其EC_(50)值为0.430 7 mg·mL^(-1)。石榕树(叶)乙酸乙酯层萃取物进行柱色谱分离后得到对甘蔗凤梨病菌有较高抑菌活性的是LMNO1bc1a活性组分,0.1 mg·mL^(-1)的LMNO1bc1a活性组分对甘蔗凤梨病菌的抑菌率为100.00%。LMNO1bc1a组分经结晶、重结晶纯化后命名为SRS-1,SRS-1经NMR和MS技术鉴定为补骨脂素(psoralen)。SRS-1对甘蔗凤梨病菌、甘蓝黑斑病菌、玉米大斑病菌、罗汉果白绢病菌、柑橘砂皮病菌、茶轮斑病菌、烟草黑胫病菌这7种植物病原真菌都有一定的抑菌活性,其EC_(50)值分别为0.018 2、0.038 8、0.040 9、0.051 2、0.080 7、0.092 7、0.115 4 mg·mL^(-1)。[结论]从石榕树中分离出抑菌活性单体化合物SRS-1,鉴定为补骨脂素,其对甘蔗凤梨病菌、甘蓝黑斑病菌、玉米大斑病菌、罗汉果白绢病菌、柑橘砂皮病菌、茶轮斑病菌、烟草黑胫病菌这7种植物病原真菌都有一定的抑菌活性。

黑龙江省延寿县驯化软枣猕猴桃品种果实品质比较分析

以2022年10月1日于黑龙江省延寿县长寿山庄采摘4种野生驯化的软枣猕猴桃品种(“果佬一号”“果佬二号”“东林一号”和“东林二号”)的果实和叶片为试材,采用多种生理指标测定方法,研究了果实品质特征,叶片、果实及植株的形态特征,以期为今后的广泛推广种植提供参考依据。结果表明:“果佬一号”和“果佬二号”果实较大,其果实平均鲜质量较高,分别为41.65 g和42.55 g。“果佬一号”和“东林二号”具有较高的可溶性糖含量,分别为6.19 mg·g^(-1)FW和6.21 mg·g^(-1)FW;“果佬二号”的维生素C含量最高为48.07μmol·g^(-1)FW;“果佬二号”和“东林二号”的可溶性固形物含量约比其他2个品种高10%;“果佬一号”和“东林一号”的可滴定酸含量约比其他2个品种高46%;“果佬二号”的固酸比最高为30.75。综上所述,4种驯化的软枣猕猴桃中,“果佬一号”产量较高且品质也较好,是综合性状均表现优良的品种;而“果佬二号”的果实红芯、适口性较好,同时也具有多个综合的优良品质,适宜推广;此外“东林一号”果实耐储运、红芯、植株颜色多变,既可用于酿酒或制作果醋,也可以用于观赏;“东林二号”果实红芯、品质好、产量适中,也较适合推广。

辽东楤木芽多糖乙酰化修饰工艺及其抗氧化活性研究

以辽东楤木芽为原料,采用超声波辅助提取法提取辽东楤木芽多糖,制备辽东楤木芽乙酰化多糖(Ac-AEBP)。以乙酰基取代度为指标,研究反应温度、反应时间和液料比(mL/g)对AEBP乙酰化修饰取代的影响。在单因素实验的基础上,运用Box-Behnken设计响应面实验对乙酰化工艺参数进行优化,并比较乙酰化修饰前后的辽东楤木芽多糖对·OH、·O^(2-)、DPPH·、ABTS+·的清除作用以及还原能力,探讨其抗氧化活性。结果表明辽东楤木芽多糖乙酰化最佳工艺条件为:反应温度52℃,反应时间4 h,液料比43∶1(mL/g),在此条件下,乙酰化辽东楤木芽多糖的取代度为0.4596。抗氧化研究表明,与未经修饰的AEBP相比,Ac-AEBP的抗氧化活性有较大的增强。该研究可为进一步开发利用辽东楤木芽提供科学依据。

浙贝母逆转恶性肿瘤耐药的研究进展

肿瘤治疗过程中,耐药的出现会大大降低治疗效果,有效逆转肿瘤细胞耐药是改善患者预后的关键。现代药理研究表明,浙贝母具有逆转肿瘤细胞耐药的作用,常用于白血病、乳腺癌、肝癌等多种恶性肿瘤的研究中。浙贝母中的化学成分可通过抑制P-糖蛋白(P-gp)的表达、减少药物外排、增加胞内药物浓度等机制,在多种肿瘤细胞中实现其逆转耐药的作用;亦可通过阻滞细胞周期诱导肿瘤细胞凋亡,进而增加肿瘤细胞内抗癌药物浓度,逆转肿瘤细胞耐药。现通过对近年来浙贝母化学成分及其逆转肿瘤细胞耐药的相关研究进行综述,以期促进浙贝母的深入研究,为后续应用于临床提供可靠的依据。

三种中草药及配伍药提取物体外抗氧化部位筛选

传统的抗癌药物多为化学药容易发生癌细胞耐药现象且副作用多,为了寻找新的抗癌药源,并探究中草药配伍能否提高原有活性,本试验选用中药材山慈菇Cremastra appendicutata(D.Don)Makino、藤梨根Actinidia arguta(Sieb.&Zucc)Planch.ex Miq、两头尖Anemone raddeana Regel,对其95%乙醇单体提取物及配伍提取物用石油醚、氯仿、乙酸乙酯、正丁醇、水萃取部分用DPPH法进行体外抗氧化活性部位筛选及抗氧化活性分析,结果发现三种中药材的乙酸乙酯层和正丁醇层提取物抗氧化活性明显高于其他层提取物,并且配伍后的中草药提取物抗氧化活性明显高于单一药材提取物。