Analysis of genes related to xylem cell wall development based on transcriptomics in Populus alba ‘Berolinensis’ tension wood

Populus alba‘Berolinensis’is a fast-growing,high-yielding species with strong biotic and abiotic stress resistance,and widely planted for timber,shelter belts and aesthetic purposes.In this study,molecular development is explored and the important genes regulating xylem forma-tion in P.alba‘Berolinensis’under artificial bending treat-ments was identified.Anatomical investigation indicated that tension wood(TW)was characterized by eccentric growth of xylem and was enriched in cellulose;the degree of ligni-fication was lower than for normal wood(NW)and oppo-site wood(OW).RNA-Seq-based transcriptome analysis was performed using developing xylem from three wood types(TW,OW and NW).A large number of differentially expressed genes(DEGs)were screened and 4889 counted.In GO and KEGG enrichment results,genes involved in plant hormone signal transduction,phenylpropanoid biosynthesis,and cell wall and secondary cell wall biogenesis play major roles in xylem development under artificial bending.Eight expansin(PalEXP)genes were identified from the RNA-seq data;four were differentially expressed during tension wood formation.Phylogenetic analysis indicated that PalEXLB1 belongs to the EXPB subfamily and that the other PalEXPs are members of the EXPA subfamily.A transcriptional regulatory network construction showed 10 transcription factors located in the first and second layers upstream of EXP,including WRKY,ERF and bHLH.RT‒qPCR analy-sis in leaves,stems and roots combined with transcriptome analysis suggests that PalEXPA2,PalEXPA4 and PalEXPA15 play significant regulatory roles in cell wall formation during tension wood development.The candidate genes involved in xylem cell wall development during tension wood formation marks an important step toward identifying the molecular regulatory mechanism of xylem development and wood property improvement in P.alba‘Berolinensis’.

Effects of Thidiazuron,basal medium and light quality on adventitious shoot regeneration from in vitro cultured stem of Populus alba×P. berolinensis

The effect of Thidiazuron （TDZ）, basal media and light quality on adventitious shoot regeneration from in vitro cultured stem of Populus albaxP berolinensis were determined to establish a high efficiency shoot regeneration system from stem explants of P. alba^P berolinensis. Stems ofPopulus alba^P berolinensis were collected from cultured shoots in vitro derived from dormancy buds of 3-year-old seedlings. The stem explants were cultured on MS medium containing 0.02-mg·L-1NAA （naphthaleneacetic acid）, and 0.1, 0.3, 0.5 and 1.0 mg·L-1 concentrations of TDZ to determine the effect of TDZ on shoot regeneration. Three basal media, i.e. MS, woody plant medium （WPM） and B5, were used to test their influences of different media on adventitious shoot regeneration. Green, red, blue and yellow plastic films in comparison with florescent light as control were used to observe their effects on shoot regeneration. The results showed that differ- ent concentrations of TDZ had an evident influence on shoot regeneration. Lower concentration of TDZ （0.1 mg·L-1） resulted in more ad- ventitious shoot regeneration and higher concentration of TDZ （〉0.1 mg·L-1） inhibited shoot regeneration. Among different media, MS medium exhibited a high efficiency for shoot regeneration, followed by WPM medium, while B5 medium inhibited shoot regeneration. Normal light and yellow light exhibited better effects on shoot regeneration, compared with other light.

Somaclonal variations as a mean for obtaining regenerants with different growth rates in poplar (<i>Populus ×berolinensis</i>Dipp.)

Adventive regenerants of Populus × berolinensis Dipp. were obtained on 1/2 MS salts with sucrose, vitamins, BA, TDZ and NAA using stem, petiole and leaf explants. They differed from each other in plantlet dimensions. More than 50 clones were produced from initial regenerants by excision and rooting of apexes and lateral shoots. Differences in stem length and thickness were observed between 200 field plants produced from in vitro plantlets. Differences in stem length were conditioned mainly due to different number of internodes and not by internodes’ lengths. Plants grown from cuttings excised from highest and smallest plants retained the abilities of mother plants to grow with different rates. It is concluded that somaclonal variability can be used for selection of fast growing poplar plants which are necessary for industrial plantations. These plants can be used for this purpose without the limitations existing for transgenic plants.

Protective mechanism of Paeoniae Radix Alba against chemical liver injury based on network pharmacology,molecular docking,and in vitro experiments

Objective:To explore and validate the potential targets of Paeoniae Radix Alba(P.Radix,Bai Shao)in protecting against chemical liver injury through network pharmacology,molecular docking technology,and in vitro cell experiments.Methods:Network pharmacology was used to identify the common potential targets of P.Radix and chemical liver injury.Molecular docking was used to fit the components,which were subsequently verified in vitro.A cell model of hepatic fibrosis was established by activating hepatic stellate cell(HSC)-LX2 cells with 10 ng/mL transforming growth factor-β1.The cells were exposed to different concentrations of total glucosides of paeony(TGP),the active substance of P.Radix,and then evaluated using the cell counting kit-8 assay,enzyme-linked immunosorbent assay,and western blot.Results:Analysis through network pharmacology revealed 13 key compounds of P.Radix,and the potential targets for preventing chemical liver injury were IL-6,AKT serine/threonine kinase 1,jun protooncogene,heat shock protein 90 alpha family class A member 1(HSP90AA1),peroxisome proliferator activated receptor gamma(PPARG),PTGS2,and CASP3.Gene Ontology(GO)enrichment analysis indicated the involvement of response to drugs,membrane rafts,and peptide binding.Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis revealed that the main pathways involved lipid and atherosclerosis and chemical carcinogenesis-receptor activation.Paeoniflorin and albiflorin exhibited strong affinity for HSP90AA1,PTGS2,PPARG,and CASP3.Different concentrations of TGP can inhibit the expression of COL-I,COL-III,IL-6,TNF-a,IL-1β,HSP-90a,and PTGS2 while increasing the expression of PPAR-γand CASP3 in activated HSC-LX2 cells.Conclusion:P.Radix primarily can regulate targets such as HSP90AA1,PTGS2,PPARG,CASP3.TGP,the main active compound of P.Radix,protects against chemical liver injury by reducing the inflammatory response,activating apoptotic proteins,and promoting the apoptosis of activated HSCs.

Development of Morus alba Reinforced Poly-Lactic Acid with Elevated Mechanical and Thermal Properties

This research investigates the mechanical and thermal properties of Morus alba combined with polylactic acid in comparison with other natural fibers. The study uses three different fiber and PLA compositions - 20%, 30%, and 40% respectively - to produce composite materials. In addition, another composite with the same fiber volume is treated with a 4% NaOH solution to improve mechanical properties. The composites are processed by twin-screw extrusion, granulation, and injection molding. Tensile strength measurements of raw fibers and NaOH-treated fibers were carried out using a single-fiber tensile test with a gauge length of 40 mm. It was observed that the NaOH surface treatment increases the resistance against tensile loading and exhibited improved properties for raw fiber strands. The diameter of the fibers was measured using optical microscopy. During this research, flexural tests, impact tests, differential scanning calorimetry (DSC), and heat deflection temperature measurements (HDT) were conducted to evaluate the mechanical and thermal properties of the developed composite samples. The results indicate that the mechanical properties of NaOH-treated Morus alba-reinforced polylactic acid outperform both virgin PLA samples and untreated Morus alba samples.

Analysis of Active Components of Hexane Extractives of Morus alba Leaves

[Objective]The aim was to explore the active compound of Morus alba leaves with the analysis of active components of hexane extractives of M.alba leaves.[Method]Antibacterial assays of hexane extractives of M.alba leaves were done and the volatile components of hexane extractives of M.alba leaves with the strongest antibacterial activity were analyzed by gas chromatography-mass spectrometry（GC-MS）.[Result]The results of inhibitory effect of hexane extractives of M.alba leaves on Staphylococcus aureus and Escherichia coli showed that hexane extractives of M.alba leaves collected in June,July and August had antibacterial activity against S.aureus and had no antibacterial effect against E.coli.Furthermore,hexane extractives of M.alba leaves collected in June had the strongest antibacterial activity against S.aureus（inhibition diameter 10.95 mm）.The hexane extractives of M.alba leaves collected in June was analyzed by GC-MS,the main volatile components were tetradecane（16.76%）,dodecane（13.20%）,diisobutyl phthalate（10.26%）,decane（9.10%）,hexadecane（8.71%）,linolenyl alcohol（7.25%）,octadecane（5.88%）,eicosane（3.26%）,dibutyl phthalate（2.59%）.[Conclusion]Linolenyl alcohol was the potential antibacterial compound.

Composition of Essential Oils of Morus alba Leaves and Chamaecyparis pisifera Outer Heartwood

[Objective] The aim was to explore the composition of essential oils of Morus alba leaves and Chamaecyparis pisifera outer heartwood. [Method] The essential oils of M. alba leaves and C. pisifera outer heartwood were analyzed by gas chromatography-mass spectrometry,respectively. [Result] Alcohols [phytol （59.47%,extracted 3 h for M. alba leaves）,α-cadinol （38.77%,extracted 6 h for C. pisifera outer heartwood）] were their first principal volatile components. Phytol was usually used as pharmaceutical raw material and α-cadinol was a kind of active organic compound with antibacterial function. [Conclusion] The essential oils of M. alba leaves and C. pisifera outer heartwood with hydrodistillation have medicinal value.

24个白桑(Morus alba L．)地方品种的遗传多样性分析

采用ISSR分子标记技术对山东、河北地区的24个白桑（Morus alba L．）地方品种资源进行了遗传多态性分析。筛选的13条ISSR引物共扩增86条扩增带，其中多态性条带63条，多态性比率为73．25％，ISSR标记遗传相似系数范围在0．6706～0．9529。通过类平均聚类（UPGMA）法分析，24份材料聚分为2大类。

Isolation and Identification of Ecliptasaponin A andEcliptasaponin B from Eclipta alba(L.)Hassk

Two new triterpenoid glucosides ecliptasaponin A(3)and ecliptasaponin B(4)were isolated together with echinocystic acid(1)and oleanolic acid(2)from Eclipta alba(L.) Hassk.Their structures were deduced as 3β,16a-dihydroxy olean-12-ene-28-oic acid-3β-O-β-D-glu-copyranoside(3)and 3β-O-[β-D-glucopyranosyl(1-4)]-β-D-glucopyranosyl-16a-hydroxy olean-12-ene-28-oic acid 28-O-β-D-glucopyranoside(4),based on spectral analysis and chemical evidences as well as results ofhydrolysis.

Two New Triterpenoid Saponins from Morina nepalensis var. alba

Two new triterpenoid saponins, monepaloside K (1) and monepaloside L (2), together with a known saponin, mazusaponin I (3), were isolated from the water-soluble part of the whole plant of Marina nepalensis var. alba Hand.-Mazz. The structures of monepalosides K and L were determined to be 3-O-α-L-arabinopyranosyl-(1-3)-α-D-xylopyranosyl siaresinolic acid (1) and 3-O-β-D-glucopyranosyl-( 1-3)-β-L-arabinopyranosy1 siaresinolic acid 28-O-β-D-glucopyranosyl-(1-6)-β-D-glucopyranoside (2) respectively, on the basis of chemical and spectroscopic evidence.

白环柄菇[Lepiota alba(Bresad.)Sacc]的生理学特性和栽培研究

白环柄菇是四川省菌物的一个新记录物种分类学地位为：担子菌亚门，层菌纲，伞菌目，伞菌科，环柄菇属。此菌无毒，可食。菌丝光锁状联合，形成大量菌核（Sclerotium），在伞菌目中是罕见的。详细报道了该菌的子实体、菌丝体的形态特征、生理特点和栽培技术。这是一种典型的高温性草菌。栽培出菇的培养料为草粉、牛粪、皮及食用菌栽培废料，瓶栽、袋栽、床栽均可，干菇生物学效率为4％-8％。

酸雨胁迫对白兰花(Michelia alba DC．)抗氧化酶活性及丙二醛(MDA)含量的影响

本文以长势良好且树龄相同的木兰科植物白兰花(Michelia alba.DC)为实验材料,采用盆栽方式,用模拟酸雨的处理方法,研究了其在模拟酸雨胁迫下的抗氧化酶系统和丙二醛(MDA)的变化规律。结果表明,pH2.0酸雨比pH3.0和pH4.0对白兰花的影响更大,到第六次喷施酸雨时,SOD、POD、CAT酶活性均低于对照;各处理MDA含量在第六次处理时达到最高值,且均与对照呈显著性差异(P<0.05)。说明酸雨胁迫对白兰花的影响较大,白兰花在酸雨胁迫下抗逆性和耐受性较弱,其结果为酸雨控制区选择城市绿化植物提供重要的现实意义。

不同光质对白芥(Sinapis alba)下胚轴愈伤组织形成和器官再生的影响(简报)(英文)

以连续白光下或黑暗中生长5天的白芥幼苗下胚轴切段为外植体(分别叫作L-外植体和D-外植体),在1mg/L NAA+2mg/L BA的MS培养基上诱导脱分化和芽的再分化。在白光、红光、蓝光和黑暗中培养,L-外植体都比D-外植体愈伤组织形成早,生长快;对于L-外植体形成愈伤组织的促进作用依次是:白光>红光>蓝光》黑暗;而对D-外植体蓝光比红光更有效。L-外植体比D-外植体芽再生的能力强得多,不同光质的作用为:白光>红光>蓝光>黑暗。

当年生桑树(Morus alba L.)果枝叶片特征与果实性状的相关性

为揭示叶果间的内在作用规律,以桑树(Morus alba L.)当年生果枝为研究对象,分析不同大小果枝上叶片特征和果实性状的差异及其二者间的相互关系。结果表明:随着果枝由小到大,果枝上的叶片数与总叶干重显著增加,而叶形指数却无显著变化;桑葚数与总桑葚干重也有显著增加,桑葚含水量无显著变化。桑葚数与叶片数、总叶干重、总叶面积及其平均桑葚干重显著正相关。研究结果证实了果实的性状与叶片特征之间存在极密切关系。当年生果枝上的桑葚数量多少取决于果枝上的叶片总数,平均桑葚的干重则与果枝上的平均叶面积显著相关。

亳州市土壤-白芍典型重金属含量分布及风险评价

以亳州市谯城区为研究区域,采集了143组白芍及根际土壤样品,分析了土壤和白芍中典型重金属Cd、Cr、Ni的含量,并采用单因子污染指数法(P_(i))、潜在生态风险指数法(RI、E_(i))和生物富集系数(BCR)评价土壤重金属污染和白芍重金属富集特征.同时,利用危害商值、靶器官毒性剂量法、致癌风险值和蒙特卡洛模拟等方法对白芍中Cd、Cr、Ni对人体健康的风险进行评估.研究结果显示,土壤中Cd、Cr、Ni的含量范围分别为0.09—0.42、24.07—117.13、26.74—62.96 mg·kg^(−1).P_(i)值显示Cd污染程度最为严重;RI值表明研究区土壤的潜在生态风险水平较低;BCR结果表明相较于Cr和Ni,白芍更容易富集Cd.人体健康风险评估结果显示,白芍中Cd、Cr、Ni的非致癌风险值均小于1,但靶器官毒性剂量法修正后的非致癌健康风险值结果高于传统的危害商值法,尤其是对于Cr造成的非致癌风险明显提高;致癌风险值在1×10^(−6)至1×10^(−4)之间,均处于可接受范围内;利用蒙特卡洛模拟进行不确定性分析,对非致癌健康风险和致癌风险的评估结果与确定性评估一致.结果表明,亳州市谯城区的土壤和白芍中重金属污染程度较低,白芍中典型重金属Cd、Cr、Ni产生的人体健康风险也处于可接受水平.

白芍(Radix paeoniae alba)提取物对中华绒螯蟹(Eriocheir sinensis)造血组织发育及造血因子Astakine的影响

血淋巴细胞在甲壳动物免疫反应中起着非常重要的作用,其主要由造血组织(Hematopoietic tissue,HPT)生成并随着开管式循环分布到身体各处。到目前为止,还没有相关研究表明甲壳动物造血组织是否受草本植物的影响。本研究以中华绒螯蟹(Eriocheir sinensis)为对象,利用荧光定量PCR、免疫荧光和组织化学等技术,初步探索了白芍(Radix paeoniae alba)提取物对中华绒螯蟹造血组织和造血因子Astakine的影响。结果显示,在原代培养的造血组织细胞中添加终浓度为0.85 mg/mL的白芍提取物后,造血因子Astakine mRNA的表达量显著上调(P<0.05)。进一步制备含1%白芍提取物的中华绒螯蟹饲料并进行饲喂实验,结果显示饲喂含白芍提取物饲料的中华绒螯蟹造血组织的Astakine mRNA的表达量较饲喂前上调了10.5倍,要高于对照组。免疫荧光结果同样表明,饲喂含白芍提取物饲料后中华绒螯蟹造血组织的Astakine蛋白荧光信号也要强于对照组。通过造血组织的形态学观察和造血组织体重百分比的检测,我们发现白芍组中华绒螯蟹造血组织含有较多卵形小叶,且造血组织体重百分比较饲喂前增加了6.64%,而对照组仅增加1.6%。免疫保护率实验显示,在脂多糖急性刺激下,白芍添加组的中华绒螯蟹的存活率(77.5%)要高于对照组(57.5%)。综上所述,白芍提取物可以促进中华绒螯蟹造血因子的表达和造血组织的发育,从而提高中华绒螯蟹的免疫调节功能。

基于古今医案云平台挖掘中药治疗肝郁型多囊卵巢综合征的用药规律

【目的】基于古今医案云平台探讨中药治疗肝郁型多囊卵巢综合征(PCOS)的用药规律。【方法】检索中国知网、维普、万方等数据库从建库至2013年12月所收录的中医药治疗肝郁型PCOS的医案,提取方药及证型数据,构建数据库,运用古今医案云平台(V2.3.7)进行肝郁证相关证型统计、中药频次统计、中药属性及功效分析、药物的关联分析与聚类分析,采用复杂网络分析挖掘其核心处方。【结果】共纳入医案处方108首,涉及药物174味,证型以肾虚肝郁为主,用药频次最高的是当归和柴胡;药物归经以归肝、脾、肾经为主,药物属性以气平、味甘最常见,药物功效以疏肝解郁为主。关联分析得到“当归-白芍”“柴胡-白芍”等17个药对;聚类分析得到4组中药聚类组合。核心处方为柴胡、白芍、当归、茯苓、甘草、菟丝子、熟地黄、香附、白术。【结论】中医药治疗肝郁型PCOS重在疏肝健脾、补益肝肾,以达气血冲任调和。

Mosquito larvicidal and ovicidal properties of Eclipta alba(L.) Hassk(Asteraceae) against chikungunya vector,Aedes aegypti(Linn.)(Diptera:Culicidae)

Objective:The present study deals with the investigation of larvicidal and ovicidal activities of benzene,hexane,ethyl acetate,methanol and chloroform leaf extract of Eclipta alba（E.alba） against dengue vector,Aedes aegypti（Ae.Aegypti）.Methods:Twenty five earlyⅢinstar larvae of Ae.aegypti was exposed to various concentrations（50-300 ppm） and was assayed in the laboratory by using the protocol of WHO 2005;the 24 h LC50 values of the E.alba leaf extract was determined by Probit analysis.For ovicidal activity,slightly modified method of Su and Mulla was performed.The ovicidal activity was determined against Ae.aegypti to various concentrations ranging from 100-350 ppm under the laboratory conditions.The egg hatch rates were assessed 48 h post treatment.Results:The LC50 values of benzene,hexane,ethyl acetate,methanol and chloroform extract of E.alba against early third instar larvae of Ae.aegypti were 151.38,165.10, 154.88,127.64 and 146.28 ppm,respectively.Maximum larvicidal activity was observed in the methanol extract followed by chloroform,benzene,ethyl acetate and hexane extract.No mortality was observed in control.Among five solvent tested the methanol extract was found to be most effective for ovicidal activity against Ae.aegypti.The methanol extracts exerted 100%mortality （zero hatchability） at 300 ppm.Conclusions:From the results it can be concluded the crude extract of E.alba was an excellent potential for controlling Ae.aegypti mosquito.

Identification of metabolites of Radix Paeoniae Alba extract in rat bile, plasma and urine by ultra-performance liquid chromatography–quadrupole time-of-flight mass spectrometry

Ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry （UPLC-Q-TOF/MS） was developed to identify the absorbed parent components and metabolites in rat bile, plasma and urine after oral administration of Radix Paeoniae Alba extract （RPAE）. A total of 65 compounds were detected in rat bile, plasma and urine samples, including 11 parent compounds and 54 metabolites. The results indicated that glucuronidation, hydroxylation and methylation were the major metabolic pathways of the components of RPAE. Furthermore, the results of this work demonstrated that UPLC-Q-TOF/MS combined with MetaboLynx？ software and mass defect filtering （MDF） could provide unique high throughput capabilities for drug metabolism study, with excellent MS mass accuracy and enhanced MSE data acquisition. With the MSE technique, both precursor and fragment mass spectra can be simultaneously acquired by alternating between high and low collision energy during a single chromatographic run.

Radial profile of sap flow velocity in mature Xinjiang poplar(Populus alba L. var. pyramidalis) in Northwest China

Estimation of the transpiration rate for a tree is generally based on sap flow measurements within the hydro-active stem xylem. In this study, radial variation of sap flow velocity（Js） was investigated at five depths of the xylem（1, 2, 3, 5 and 8 cm under the cambium） in three mature Xinjiang poplar（Populus alba L. var. pyramidalis） trees grown at the Gansu Minqin National Studies Station for Desert Steppe Ecosystem from May to October 2011. Thermal dissipation probes of various lengths manufactured according to the Granier＇s design were installed into each tree for simultaneous observation of the radial patterns of Js through the xylem. The radial patterns were found to fit the four-parameter GaussAmp equation. The peak Js was about 27.02±0.95 kg/（dm2？d） at approximately 3 to 5 cm deep from the cambium of the three trees,and the lowest Js appeared at 1 cm deep in most of the time. Approximately 50% of the total sap flow in Xinjiang poplar occurred within one-third of the xylem from its outer radius, whereas 90% of the total sap flow occurred within two-fifth of the xylem. In addition, the innermost point of the xylem（at 8-cm depth）, which appeared as the penultimate sap flow in most cases during the study period, was hydro-active with Js,8 of 7.55±3.83 kg/（dm2？d）. The radial pattern of Js was found to be steeper in midday than in other time of the day, and steeper diurnal fluctuations were recorded in June, July and August（the mid-growing season）. Maximum differences between the lowest Js（Js,1 or Js,8） and the highest Js（Js,3 or Js,5） from May through October were 12.41, 17.35, 16.30, 18.52, 12.60 and 16.04 g/（cm2？h）, respectively. The time-dependent changes of Js along the radial profile（except at 1-cm depth） were strongly related to the reference evapotranspiration（ET0）. Due to significant radial variability of Js, the mean daily sap flow at the whole-tree level could be over-estimated by up to 29.69% when only a single probe at depth of 2 cm was used. However, the accuracy of the estimation of sap flow in Xinjiang poplar could be significantly improved using a correction coefficient of 0.885.