Molecular cloning,characterization and promoter analysis of LbgCWIN1 and its expression profiles in response to exogenous sucrose during in vitro bulblet initiation in lily

Lily(Lilium spp.) is an important ornamental flower, which is mainly propagated by bulbs. Cell wall invertases(CWINs), which catalyze the irreversibly conversion of sucrose into glucose and fructose in the extracellular space, are key enzymes participating in sucrose allocation in higher plants. Previous studies have shown that CWINs play an essential role in bulblet initiation process in bulbous crops, but the underlying molecular mechanism remains unclear. Here, a CWIN gene of Lilium brownii var. giganteum(Lbg) was identified and amplified from genomic DNA. Quantitative RT-PCR assays revealed that the expression level of LbgCWIN1 was highly upregulated exactly when the endogenous starch degraded in non-sucrose medium during in vitro bulblet initiation in Lbg. Phylogenetic relationship, motif, and domain analysis of LbgCWIN1 protein and CWINs in other plant species showed that all sequences of these CWIN proteins were highly conserved. The promoter sequence of LbgCWIN1 possessed a number of alpha-amylase-, phytohormone-, light-and stress-responsive cis-elements. Meanwhile, β-glucuronidase(GUS) assay showed that the 459 bp upstream fragment from the translational start site displayed maximal promoter activity. These results revealed that LbgCWIN1 might function in the process of in vitro bulblet initiation and be in the response to degradation of endogenous starch.

百合总皂苷提取工艺及抗抑郁活性研究

本文研究百合总皂苷最佳提取工艺,继而采用AB-8大孔吸附树脂富集纯化百合总皂苷,并测试其抗抑郁活性。采用正交实验,考察乙醇浓度、提取次数、固液比及提取时间四个因素对该工艺的影响;采用小鼠悬尾实验和小鼠强迫游泳实验评价百合总皂苷的抗抑郁活性。确立了百合总皂苷的最佳提取工艺为:提取时间3h,乙醇浓度80%,固液比1∶10,提取次数为3次。在最佳提取工艺条件下,总皂苷提取率为1.24%。经过AB-8大孔树脂富集纯化总皂苷的含量达到62%以上。药理实验表明,百合总皂苷中剂量、小剂量能明显缩短小鼠悬尾的不动时间和游泳时间(P<0.05或者P<0.01),百合富集纯化的总皂苷部位具有较好的抗抑郁活性。

HPLC法测定百合、卷丹、细叶百合中3种甾体皂苷的含量

目的 HPLC法同时测定百合、卷丹、细叶百合中3种甾体皂苷的含量。方法采用HPLC法,色谱柱为Akzonobel Kromasil C18(250 mm×4.6 mm,5μm),流动相为乙腈-0.1%磷酸,检测波长210 nm,流速1.0mL/min。结果 (25R)-26-O-(β-D-吡喃葡萄糖)-呋甾烷-5-烯-3β,22R,26-三羟基-3-O-α-L-鼠李糖-(1→2)-β-D-吡喃葡萄糖-(1→4)-β-D-吡喃葡萄糖苷(简称皂苷1)在2～12μg呈良好线性关系(r=0.9998);(25R)-螺甾烷-5-烯-3β-O-α-L-鼠李糖-(1→2)-O-[β-D-葡萄糖-(1→6)]-β-D-吡喃葡萄糖苷(简称皂苷2)在3.125～18.75μg呈良好线性关系(r=0.9999);(25R)-3β,17α-二羟基-5α-螺甾烷-6-酮-3-O-α-L-鼠李糖-(l→2)-O-[α-L-阿拉伯糖-(l→3)]-β-D-吡喃葡萄糖苷(简称皂苷3)在1.125～9μg呈良好线性关系(r=0.9997)。皂苷1、2、3的平均回收率分别为97.72%、99.27%、98.96%,RSD分别为1.96%、1.78%、1.31%。结论所建立的HPLC法简便快速、准确度高,可用于百合药材的质量控制。

百合花色素性能研究

从百合花中提取出了橙黄色素,试验了该色素的溶解性能和光、热、pH值及盐类物质对色素水溶液稳定性的影响。结果表明:百合花色素着色力强,稳定性好。百合花是一种有待开发、具有较高经济价值的新的天然色素资源。

卷丹和百合化学成分与提取方法研究进展

介绍了近十年来,卷丹和百合化学成分和提取方法方面的研究进展,主要集中在甾体皂苷、多糖和秋水仙碱;甾体皂苷的提取方法有醇提—大孔树脂吸附法、醇提—正丁醇萃取法;多糖的提取方法有水提醇沉法和复合酶法;秋水仙碱的提取方法有有机溶剂提取法和超临界二氧化碳流体萃取法。

紫外可见分光光度法测定不同产地龙牙百合中皂苷含量

为建立龙牙百合总皂苷的测定方法,并确定不同产地龙牙百合中皂苷的含量,以便控制龙牙百合药材的质量,采用香草醛-高氯酸显色,在535 nm处,利用紫外可见分光光度法测定了10个批次的龙牙百合中总皂苷的含量。结果表明,10个不同产地龙牙百合中皂苷的含量,分别是:8.39%、8.11%、8.24%、7.62%、7.38%、9.02%、8.83%、8.87%、9.21%、9.17%,RSD分别为:1.22%、0.87%、0.93%、1.32%、0.86%、1.37%、0.94%、1.39%、1.55%、1.61%。结果表明,该法准确、可靠、稳定性好,可作为龙牙百合皂苷定量分析的有效方法。

产于浙江温岭的百合属一新变种——巨球百合

报道了发现于浙江温岭的百合属Lilium一新变种——巨球百合Lilium brownii F、E．Brown e Miellez var．giganteum G．Y．Li＆Z．H．Chen。本变种与原变种野百合Lilium brownii不同在于：鳞茎特大，直径10～12cm，鳞片100余枚，花冠淡黄色，外面带紫色；通常5～8朵。

龙牙百合核型分析

采用染色体常规压片方法对龙牙百合根尖细胞进行了染色体核型分析,结果表明,龙牙百合的染色体数目为2n=24,染色体核型公式为2n=2x=24=4m(SAT)+2sm+10st(2SAT)+8t,核型分类属于3B类型,核型不对称系数As.K为76.80%,染色体相对长度系数I.R.L=6L+4M2+10M1+4S。

平贝母等3种中药材铅、镉、砷、汞含量的研究

对中药材平贝母的铅、镉、砷、汞含量测定的方法进行了研究。样品经HNO3-HClO4 或H2 O2 进行消解 ,用原子吸收和原子荧光进行检测。回收率 :92 0 %～ 99 7% ;精密度 :2 3 %～ 4 1%。并用该方法对全国主产区的平贝母、百合和两头尖样品的铅、镉、砷、汞的含量进行了测定。结果表明 :这 3种中药材均含铅、镉、砷、汞 ,但参考粮食和蔬菜的国家标准未超标。

抑制鲜龙牙百合成分提取物褐变的分析

对鲜龙牙百合(Lilium brownii var.viridulum Baker)成分提取中的褐变处理方法进行了研究。分别采用乙醇提取法和维生素C提取法抑制提取物的褐变,确定了用乙醇提取法抑制褐变的最佳乙醇体积分数为80%;用维生素C提取法抑制褐变的最佳维生素C浓度为0.6 g/L,并证实了提取物在后续保存过程中,只需采用合适的抑制剂浓度并置于冰箱即可,无需其他特殊处理。

Integrative analysis of metabolome and transcriptome provides new insights into functional components of Lilii Bulbus

Objective:Lilium brownii var.viridulum(LB)and L.lancifolium(LL)are the main sources of medicinal lily(Lilii Bulbus,Baihe in Chinese)in China.However,the functional components of these two species responsible for the treatment efficacy are yet not clear.In order to explore the therapeutic material basis of Lilii Bulbus,we selected L.davidii var.willmottiae(LD)only used for food as the control group to analyze the differences between LD and the other two(LB and LL).Methods:Metabolome and transcriptome were carried out to investigate the differences of active components in LD vs LB and LD vs LL.Data of metabolome and transcriptome was analysed using various analysis methods,such as principal component analysis(PCA),hierarchical cluster analysis(HCA),and so on.Differentially expressed genes(DEGs)were enriched through KEGG and GO enrichment analysis.Results:The PCA and HCA of the metabolome indicated the metabolites were clearly separated and varied greatly in LL and LB contrasted with LD.There were 318 significantly differential metabolites(SDMs)in LD vs LB group and 298 SDMs in LD vs LL group.Compared with LD group,the significant up-regulation of steroidal saponins and steroidal alkaloids were detected both in LB and LL groups,especially in LB group.The HCA of transcriptome indicated that there was significant difference in LB vs LD group,while the difference between LL and LD varied slightly.Additionally,47540 DEGs in LD vs LB group and 18958 DEGs in LD vs LL group were identified.Notably,CYP450s involving in the biosynthesis of steroidal saponins and steroidal alkaloids were detected,and comparing with LD,CYP724,CYP710A,and CYP734A1 in LB and CYP90B in LL were all up-regulated.Conclusion:This study suggested that steroidal saponins and steroidal alkaloids maybe the representative functional components of Lilii Bulbus,which can provide new insights for Lilii Bulbus used in the research and development of classic famous formula.