Identifying potential flavonoid biosynthesis regulator in Zanthoxylum bungeanum Maxim. by genome-wide characterization of the MYB transcription factor gene family

Plant MYB transcription factors(TFs) play crucial roles in regulating the biosynthesis of flavonoids but current analysis on their role in Zanthoxylum bungeanum Maxim.(ZBM) is far from comprehensive. In this study, we identified 270 MYB genes in ZBM and divided them into four subfamilies. The R2 R3-MYB(ZbMYB) category contained 251 genes and was classified into 33 subfamilies according to their phylogenetic results and sequence similarity. These subfamilies included 24 subgroups containing both MYBs of ZBM plants and AtMYBs, and nine subgroups containing only ZBM MYBs or AtMYBs. ZbMYBs with similar functions clustered into the same subgroup, indicating functional conservation. The subcellular localization analysis predicted that most ZbMYB genes were found in the nucleus. The transposed duplications appeared to play a major role in the expansion of the MYB gene family in ZBM. Through phylogenetic analysis and transcriptome profiling, it was found that 28 ZbMYB genes may regulate the biosynthesis of flavonoids in ZBM, and these genes expression presented distinct temporal and spatial expression patterns. In different fruit development stages of ZBM, the expression patterns of EVM0042160 and EVM0033809 genes obtained by qRT-PCR analysis are very similar to the flavonoid and anthocyanin content curves in ZBM. Further correlation analysis showed that the content of flavonoids in different fruit development stages and the transcript abundance levels of 28 ZbMYB genes have different degrees of correlation relationship. These results indicated that the ZbMYB genes might be involved in the flavonoid metabolic pathway. This comprehensive and systematic analysis of MYB family genes provided a solid foundation for further functional analysis of MYB TFs in ZBM.

Cytotoxicity and enhancement activity of essential oil from Zanthoxylum bungeanum Maxim.as a natural transdermal penetration enhancer

The aim of this present study is to investigate the effect of Zanthoxylum bungeanum oil （essential oil from Z. bungeanum Maxim.） on cytotoxicity and the transdermal permeation of 5-fluorouracil and indomethacin. The cy- totoxicity of Z. bungeanum oil on dermal fibroblasts and epidermal keratinocytes was studied using an MTT （3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide） assay. The rat skin was employed to determine the percutaneous penetration enhancement effect of Z. bungeanum oil on hydrophilic and lipophilic model drugs, i.e., 5-fluorouracil and indomethacin. The secondary structure changes of the rat stratum comeum （SC） were determined using attenuated total reflectance-Fourier transform infrared spectroscopy （ATR-FTIR）, and saturated solubilities and SC/vehicle partition coefficients of two model drugs with and without Z. bungeanum oil were also measured to un- derstand its related mechanisms of action. It was found that the half maximal inhibitory concentration （ICs0） values of Z. bungeanum oil were significantly lower in HaCaT and CCC-ESF-1 cell lines compared to the well-established and standard penetration enhancer Azone. The Z. bungeanum oil at various concentrations effectively facilitated the percutaneous penetration of two model drugs across the rat skin. In addition, the mechanisms of permeation en- hancement by Z. bungeanum oil could be explained with saturated solubility, SC/vehicle partition coefficient, and secondary structure changes of SC.

Essential oil from Zanthoxylum bungeanum Maxim. and its main components used as transdermal penetration enhancers: a comparative study

Our previous studies had confirmed that the essential oil from Zanthoxylum bungeanum Maxim. （Z. bungeanum oil） could effectively enhance the percutaneous permeation of drug molecules as a natural transdermal penetration enhancer. The aim of the present study is to investigate and compare the skin penetration enhancement effect of Z. bungeanum oil and its main components on traditional Chinese medicine （TCM） active components. Toxicities of Z. bungeanum oil and three selected terpene compounds （terpinen-4-ol, 1,8-cineole, and limonene） in epidermal keratinocytes （HaCaT） and dermal flbroblast （CCC-ESF-1） cell lines were measured using an MTT （3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide） assay. Five model drugs in TCM external preparations, namely osthole （OT）, tetramethylpyrazine （TMP）, ferulic acid （FA）, puerarin （PR）, and geniposide （GP）, which were selected based on their lipophilicity denoted by IogKo^w, were tested using in vitro permeation studies in which vertical Franz diffusion ceils and rat abdominal skin were employed. The secondary structure changes of skin stratum corneum （SC） and drug thermodynamic activities were investigated to understand their mechanisms of action using Fourier transform infrared （FTIR） spectroscopy and saturation solubility studies, respectively. It was found that Z. bungeanum oil showed lower toxicities in both HaCaT cells and CCC-ESF-1 cells compared with three terpene compounds used alone. The enhancement permeation capacities by all tested agents were in the following increasing order： terpinen-4-ol=1,8-cineole〈limonene〈Z, bungeanum oil. The mechanisms of permeation enhancement suggested that these enhancers promoted the skin permeation of drugs mainly by affecting SC lipids. These results indicated that Z. bungeanum oil exhibited better performance in enhancing the skin permeation of active components in TCM preparations.

Transdermal permeation of Zanthoxylum bungeanum essential oil on TCM components with different lipophilicity

Objective:To investigate the percutaneous penetration enhancement effect of essential oil from Zanthoxylum bungeanum Maxim.(Z.bungeanum oil)on active components in externally-applied traditional Chinese medicines.Methods:Five model drugs,geniposide,puerarin,ferulic acid,tetramethylpyrazine,and osthole,were chosen based on their lipophilicity and tested using in vitro transdermal permeation studies consisting of Franz diffusion cells and full thickness rat abdominal skin.Scanning electron microscopy was employed to observe the morphological changes of rat skin tissue after treatment with Z.bungeanum oil.The molecular interactions between the oil and the polar head groups in stratum corneum(SC)lipids were monitored using molecular dynamic simulation,and the SC/vehicle partition coefficients and saturation solubilities of the selected model drugs treated with and without the oil were also determined to ascertain its mechanisms of action.Results:As oil concentration increased,the log ERflow trended toward a negative linear relationship with the lipophilicity of drugs.After treatment with Z.bungeanum oil,a mild lifting up and wrinkle on the SC surface were observed,and appeared to become more pronounced as oil concentration increased.There was no significant difference between the control and the Z.bungeanum oil at different concentrations in terms of saturation solubility of GP,while saturation solubilities of the 4 other drugs gradually increased as oil concentration increased.The oxygen-containing constituents in Z.bungeanum oil,such as terpinen-4-ol and 1,8-cineole,which accounted for 57.95%of total oil,could form stable hydrogen bonds with the polar head group of ceramide 3.Conclusion:Z.bungeanum oil facilitated transdermal permeation of drugs with different lipophilicity,including the extremely hydrophilic and lipophilic drugs,whereas it exhibited greater enhancement activity for strongly hydrophilic drugs.The mechanisms of transdermal permeation enhancement by the oil could be explained with SC/vehicle partition coefficient,saturation solubility,and the interactions with SC lipids.

花椒病虫害绿色防控技术研究进展

花椒(Zanthoxylum bungeanum Maxim.)经济价值较高,在中国广泛种植,但其生长过程中易遭受病虫危害,科学的防治技术措施对花椒的绿色发展至关重要。从生态调控技术、生物防治技术、理化诱控技术、科学用药技术4个方面对花椒病虫害绿色防控技术进行综述,以期为生产上花椒病虫害绿色防控提供参考。

花椒籽对蛋鸡生产性能、血清指标及蛋品质的影响

试验旨在研究饲粮中添加花椒籽对蛋鸡生产性能、血清指标和蛋品质的影响。试验选择42周龄的京粉6号蛋鸡360只,随机分为2组,每组6个重复,每个重复30只。对照组(CON组)饲喂基础饲粮,试验组(ZBMS组)饲喂使用2%花椒籽替代基础饲粮中的玉米的饲粮。预试期7 d,正式试验期28 d。结果显示:与CON组相比,ZBMS组第1~14、15~28、1~28 d的平均蛋重显著提高(P<0.05),软破蛋率显著降低(P<0.05);ZBMS组蛋鸡血清谷草转氨酶(AST)活性及白细胞介素-1β(IL-1β)、干扰素-γ(IFN-γ)水平均显著降低(P<0.05),白细胞介素-4(IL-4)水平、总抗氧化能力(T-AOC)均显著提高(P<0.05)。与CON组相比,ZBMS组第28 d时的蛋黄色泽显著提高(P<0.05);ZBMS组鸡蛋中棕榈油酸、亚油酸、α-亚麻酸、山嵛酸和二十二碳六烯酸(DHA)的含量均显著提高(P<0.05),肉豆蔻酸、十七碳酸、硬脂酸、反油酸、油酸、二十碳一烯酸、附子酯酸和花生三烯酸的含量均显著降低(P<0.05)。研究表明,使用2%花椒籽替代玉米可增强蛋鸡免疫力和抗氧化能力,提高蛋重,降低软破蛋率,提升蛋黄色泽和蛋黄比率,增加鸡蛋中多不饱和脂肪酸的含量。

甘肃陇南不同种类花椒籽的营养成分对比研究

文章旨在以甘肃省陇南市文县九月青花椒籽、红花椒籽、无刺嫁接红花椒籽、为研究对象,采用凯氏定氮法、色谱法等测定花椒籽中蛋白、挥发油等主要营养成分的含量,评价3个不同种类花椒籽的饲用价值。结果表明,经过品种改良后,无刺嫁接花椒籽中的水分、总灰分和粗纤维含量下降,粗蛋白和粗脂肪含量增加;无刺嫁接红花椒籽中粗脂肪、粗蛋白和挥发油含量显著高于九月青花椒籽和红花椒籽(P<0.05)。无刺嫁接红花椒籽中α-亚油酸、棕榈酸、硬脂酸、顺油酸、顺亚油酸和十五烷酸中含量显著高于九月青花椒籽(P<0.05)。无刺嫁接红花椒籽中的天冬氨酸、谷氨酸、精氨酸、甘氨酸、亮氨酸、丙氨酸、丝氨酸、苯丙氨酸、苏氨酸、缬氨酸、异亮氨酸、脯氨酸、组氨酸含量和氨基酸总量均显著高于九月青花椒籽和红花椒籽中的氨基酸含量(P<0.05)。无刺嫁接红花椒籽所含必需氨基酸中异亮氨酸、赖氨酸、蛋氨酸、苏氨酸均低于FAO/WHO的推荐值,且只在无刺嫁接红花椒籽中检测出蛋氨酸,其中亮氨酸、苯丙氨酸+酪氨酸和缬氨酸均高于FAO/WHO的推荐值。综上所述,3个不同种类的花椒籽中各组成成分种类和含量有所不同,其中无刺嫁接花椒籽富含多种营养成分,具有良好的饲用开发价值。

大红袍花椒对流-辐射并联干燥BP神经网络模拟与品质研究

为探究热风干燥、热风-红外和热风-微波对流-辐射并联干燥对大红袍花椒干燥特性及品质的影响,实验分析了不同温度、装载量、干燥功率等条件下的大红袍花椒干燥曲线特征,运用BP神经网络进行拟合,并采用感官评价与GC-MS对3种干燥方式干制大红袍花椒挥发油进行分析。结果表明,热风-红外并联干燥的恒速期干燥速率高于热风干燥一个数量级,热风-微波并联干燥时长最短,出现二次升速且降速期不明显;升温、减少装载量或加大微波功率均有利于提高干燥速率和缩短干燥时间。BP神经网络的相关系数R值均能达到0.985以上,均方误差最低可达1.010×10^(-4),平均相对误差值E为4.55%,可很好地描述大红袍花椒的干燥动力学特性,预测大红袍花椒干燥过程含水率准确且迅速。3种干燥方式挥发油分别鉴定出40、39、28种化学成分,热风-微波并联干燥获取大红袍花椒挥发油中烯烃类化合物相对含量最大,但色泽和口感差,热风干燥花椒色泽最佳,挥发油中化合物种类最多,热风-红外并联干燥既缩短了大红袍花椒干燥时间,又能保持其良好品质,为较佳选择。

不同温湿度大红袍花椒对流辐射干燥试验研究

为对比大红袍花椒热风干燥、热风-红外与热风-微波联合干燥的特性和品质,揭示对流辐射联合干燥大红袍花椒的干燥过程,指导大红袍花椒生产实践。通过薄层干燥试验,研究三种干燥方式在不同温度(50℃、60℃、70℃)和相对湿度(10%、30%、50%)条件下的干燥曲线和有效水分扩散系数,结合Weibull函数的尺度参数α、形状参数β及估算有效水分扩散系数进行干燥动力学分析,采用扫描电子显微镜(SEM)观察干制花椒油苞结构,提取挥发油进行气相色谱质谱(GC-MS)分析。结果表明:热风干燥时间最长,升温降湿有利于提高热风干燥速率、缩短干燥时间,但对热风-红外和热风-微波干燥影响较小;Weibull函数能很好地模拟三种干燥方式,α随干燥条件变化明显,β>1,水分迁移是由物料表面和内部共同控制,估算水分扩散系数变化范围分别为1.303×10^(-7)~2.815×10^(-7)m^(2)/min、7.646×10^(-7)~9.628×10^(-7)m^(2)/min、2.200×10^(-6)~2.778×10^(-6)m^(2)/min,且与温湿度变化相关;热风干燥和热风-红外干燥能较好地保留花椒油苞,热风-微波干燥花椒油苞更为饱满;热风-微波干燥后花椒挥发油提取量最高(7.5%),烯烃类物质相对含量最大,热风干燥花椒提取挥发油含量最低,但挥发油中化合物种类最多。热风-红外联合干燥在缩短干燥时间和保持品质方面是较好的选择。

红花椒风味品质的影响因素研究

目的:考察温度、光照、氧气3个因素对红花椒风味物质的综合影响。方法:采用HPLC测定不同影响因素下花椒中羟基-α-山椒素、羟基-β-山椒素、羟基-γ-山椒素、羟基-ε-山椒素等麻味物质含量;采用GC测定不同影响因素下花椒中芳樟醇、柠檬烯、乙酸芳樟酯等香味物质含量;采用极坐标对不同因素对风味物质的影响进行综合强度分析。结果:4℃和25℃时麻味物质含量高且差异较小,4℃时香味物质含量最高;遮光条件下麻味和香味物质含量保存最佳;隔氧条件下,随着储存时间的延长,麻味和香味物质损失较慢;结合强度分析图得出红花椒受到影响的因素主要包括氧气、温度和光照,其中氧气对其影响最显著。结论:对红花椒风味品质的影响因素按照重要性排序为氧气>温度>光照,隔氧、遮光、4℃是最佳条件。该研究为花椒调味品加工、运输和储存提供了参考依据。

花椒精油提取方法、成分分析、生物活性及实际应用研究进展

花椒是我国广泛应用的八大调味香料之一,花椒精油是从花椒果皮或籽中提取出来的一种具有芳香气味的浅黄绿色油状液体。其化学成分复杂,通常由萜类、烯类、醛类、酮类、醇类等化学物质组成;药理作用广泛,具有抗氧化、抑菌、镇痛、消炎、驱虫等功效,被广泛应用在食品、医药、化妆品等领域。文章阐述了花椒精油的不同提取方法及其优缺点,分析了花椒精油的化学成分以及生物活性,并通过对国内外研究现状进行分析,具体阐述了花椒精油在果蔬保鲜、肉制品保鲜以及生物医药等领域的应用并对其研究进行了展望,以期为花椒精油的产业化发展提供理论参考。

花椒品质评价研究进展

花椒是一种传统药食同源中药,具有“温中止痛,杀虫止痒”的功效,现代临床上具有抗炎、抗肿瘤、镇痛、抗氧化、麻醉等功效.花椒品质的优劣直接决定了花椒的药用价值和高附加值产品开发,因此花椒品质评价研究一直是科研人员关注的热点之一.从色泽、香味、麻味、谱效相关等方面对花椒品质评价的研究现状和方法技术进展进行了介绍,具体地介绍了花椒色泽的感官评价特征分析技术,花椒香味和麻味成分的光谱、色谱及电化学等分离分析技术,花椒成分与功能功效的“谱效相关”研究,花椒品质评价研究进展和发展趋势等,以期为花椒品质评价、资源保障、高价值产品研究提供参考.

青海省东部农业区花椒种质资源调查

为调查青海省的花椒种质资源,文章调查收集了青海省东部农业区7个县(区)的花椒种质资源37份,并对其植物学表型性状、株型、枝条、叶片、果实等进行调查。结果显示,37份花椒种质资源19项表型性状指标的变异系数在7.26%~59.82%之间,整体变异系数平均为18.17%,变异系数最大的指标为单个果穗果实数量,为59.82%;出皮率的变异系数最小,为7.26%。花椒枝条表型性状的变异系数在13.43%~21.94%之间,叶片的表型性状变异系数在14.14%~19.73%之间,果实表型性状的变异系数在7.26%~59.82%之间。37份样本的19个表型性状分析结果,在一定程度上反映了青海省东部农业区花椒种质资源的多样性,为今后深入研究其遗传多样性及开发利用奠定了基础。

基于蛋白质组学探讨羟基-α-山椒素对糖尿病心肌病的作用机制

目的基于蛋白质组学检测技术,探讨羟基-α-山椒素(hydroxy-α-sanshool,SAN)治疗糖尿病心肌病(diabetic cardiomyopathy,DCM)的作用机制。方法高脂饲料喂养及腹腔注射链脲佐菌素(streptozotocin,STZ)联合使用建立DCM模型。分为对照组(CON组)、DCM组和SAN治疗组(DCM+SAN组)。小动物心脏超声及病理染色评价小鼠心脏功能及组织形态,蛋白组学技术推测SAN对DCM的作用机制,Western blot实验验证cAMP/PKA信号通路关键蛋白表达水平。结果心脏超声与病理染色显示SAN对DCM小鼠心脏有保护作用。DCM+SAN组与DCM组进行蛋白组学检测,共鉴定160个差异蛋白,127个蛋白表达上调,33个蛋白表达下调;GO二级功能注释显示细胞过程、分子结构和细胞结构等功能;KEGG富集分析显示cAMP信号通路富集最为丰富;蛋白互作网络显示以PKA为中心节点与多种蛋白在cAMP信号通路中有互作关系。Western blot验证DCM组的cAMP和PKA蛋白相对表达量均明显低于CON组(P<0.05);DCM+SAN组的cAMP和PKA蛋白相对表达量均明显高于DCM组(P<0.05)。结论SAN对糖尿病心功能有保护作用,通过cAMP信号通路发挥作用。

不同海拔下花椒叶片差异表达基因的结构变异

[目的]探明不同海拔梯度对花椒基因表达的影响,为研究花椒遗传多样性、代谢调控及适应不同海拔生境胁迫的分子机制提供依据。[方法]以贵州省贞丰县板贵乡不同海拔(1054、821和645 m)下的花椒叶为试验材料,采用转录组测序的生物信息学方法对其差异表达基因的可变剪接(AS)和SNP/InDel变异位点进行解析,挖掘花椒适应环境的新转录本。[结果]在3个不同海拔的花椒叶片中,有2619个基因发生了可变剪切事件,主要为外显子跳跃(Skipped exon,SE)和互斥外显子(Mutually exclusive exons,MXE)2种类型,其中SE最多,且在高海拔的对比组中发生可变剪接的数量显著增多;鉴定出748331个SNP和87402个InDel位点,SNP类型中转换种类高于颠换种类,在外显子区分布最多,达40.0%。InDel位点在内含子区分布最多,达28.6%;发掘出9363个新转录本,其中有5648个在7大数据库中被注释。KEGG代谢途径分析发现,花椒叶片发生可变剪接的功能基因主要参与代谢途径、次生代谢物的生物合成和辅助因子的生物合成途径;而新转录本的KEGG代谢途径主要富集于光合作用、苯丙素生物合成、异黄酮生物合成、类黄酮生物合成和萜类生物合成等通路。[结论]本研究丰富了对不同海拔条件下花椒基因结构变异的认识,为后续研究花椒遗传多样性、代谢调控及响应环境胁迫的分子机制奠定基础。

蛇床子、花椒、白矾、苦参加减辨治对外阴阴道炎阴道微环境影响

目的:观察蛇床子、花椒、白矾、苦参加减辨治对外阴阴道炎临床疗效和对阴道微环境的影响。方法:选取120例外阴阴道炎患者随机分为观察组60例和对照组60例,对照组采用硝呋太尔制霉菌素阴道软膏治疗,观察组采用蛇床子、花椒、白矾、苦参加减辨治药方进行熏洗治疗。比较两组治疗前后临床疗效和阴道微环境变化情况。结果:观察组临床疗效总有效率(95.0%)高于对照组(81.7%),差异具有统计学意义(P<0.05);治疗后,两组阴道pH值、阴道清洁度、阴道菌群密集度均优于治疗前(P<0.05),且观察组优于对照组(P<0.05),两组阴道过氧化氢阳性率、白细胞酯酶阳性率均低于治疗前(P<0.05),且观察组低于对照组(P<0.05)。结论:中药蛇床子、花椒、白矾、苦参加减辨治方对外阴阴道炎有良好疗效,同时可有效改善阴道微环境。

果实膨大期不同浓度GA_(3)处理对大红袍花椒果实品质影响

施加GA_(3)等植物生长调节剂已成为提高果实品质的重要栽培管理手段,但施加GA_(3)对花椒果实品质的影响目前仍不清楚。为研究不同浓度GA_(3)处理在果实膨大期对大红袍花椒果实的外在品质和内在品质的影响,筛选出最优GA_(3)喷施浓度。选择大红袍花椒为研究对象,在花椒果实膨大期喷施浓度为25 mg/L,50 mg/L,75 mg/L和100 mg/L的GA_(3),待果实成熟采收后,分别测定果粒的纵径、横径、百粒重、果皮油囊数量、总挥发油、酰胺类化合物、不挥发性乙醚萃取物以及总黄酮指标。结果表明,坐果期喷施75 mg/L GA_(3)时综合效果最好,果实纵径,横径,百粒重以及果皮油囊数量4项外在品质指标分别提升14%、11%、12%和35%。果皮总挥发油、麻味物质、不挥发性乙醚萃取物和总黄酮4项内在品质指标会显著提升33%、27%、13%和11%。本研究筛选出了提升花椒果实品质的最佳GA_(3)处理浓度,为花椒栽培管理提供了新的见解和思路。

花椒BAHD酰基转移酶基因家族鉴定及表达差异分析

麻味物质是评价花椒(Zanthoxylum bungeanum Maxim.)品质的重要特征性成分之一,为了挖掘花椒麻味物质合成途径酰化反应的候选基因,本研究基于花椒全基因组数据,通过生物信息学方法,对花椒BAHD酰基转移酶基因家族进行了研究。研究结果共鉴定出50个花椒BAHD酰基转移酶基因家族成员,这些基因可分为两大亚家族,均编码不稳定亲水性蛋白;亚细胞定位分析表明大部分花椒BAHD成员主要定位于细胞质,其它成员定位于细胞核、叶绿体等细胞器中。基因表达分析显示,BAHD酰基转移酶基因在花椒果皮发育各阶段均有特异性的表达,且在果梗、刺、茎和花中表达高于种子和叶。不同花椒品种间,即使相同基因也表现出表达差异,甚至在某些品种中未检测到表达。此研究为花椒麻味物质合成途径提供了新的基因资源,并将为其它高等植物BAHD酰基转移酶基因家族的研究提供参考。

气相色谱-串联质谱法测定花椒中多种农药残留

本研究采用气相色谱-串联质谱法对花椒中的多种农药残留进行测定。结果表明,10种农药在0.002~0.400 mg·L^(-1)范围内线性关系良好,相关系数大于0.99;在5个浓度(0.01 mg·kg^(-1)、0.02 mg·kg^(-1)、0.05 mg·kg^(-1)、0.10 mg·kg^(-1)和0.20 mg·kg^(-1))添加水平下的平均回收率为63%~128%,相对标准偏差为1.2%~9.7%,符合农药残留分析标准。该方法具有准确度高、精密度好、线性范围宽等优点,可为花椒中农药残留的检测提供有力支持。

近红外光谱快速无损检测花椒中麻素及精油含量

利用近红外光谱法结合偏最小二乘法构建定量分析模型,同时实现花椒中麻素及精油含量的快速无损检测。取92份花椒样品,采集近红外光谱用于模型的建立及验证。采用二阶导数进行光谱预处理,在优选的波数范围和主因子下,建立的麻素及精油定量模型的相关系数分别为0.9665和0.9778,预测均方差分别为0.470和0.413,在外部验证中相对偏差均小于5%,说明所建模型预测性能良好,可用于花椒品质的快速评价。