紫花地丁HPLC指纹图谱研究

目的:建立紫花地丁药材HPLC指纹图谱,提供药材质量控制的可靠方法。方法:采用HPLC方法,以Agi-lent C18(4.6 mm×250 mm,5μm)为色谱柱,甲醇-0.5%醋酸水溶液进行梯度洗脱;检测波长353 nm,流速1.0mL/min。结果:检测了12批不同来源的紫花地丁药材,确立了18个共有峰,建立了紫花地丁对照指纹图谱,计算各被测样品的HPLC指纹图谱的整体相似度,并指认了菊苣苷、七叶内酯、东莨菪素、早开堇菜苷4个特征峰,比较了上述成分在不同药材中的含量。结论:所建立的指纹图谱具有良好的精密度、重现性和稳定性,可作为紫花地丁药材质量控制标准。     

Patterns of flower morphology and structural changes during interconversion between chasmogamous and cleistogamous flowers in Viola philippica

Aims Viola philippica is a species with a typical chasmogamous-cleistogamous (CH-CL) mixed breeding system. It provides a flower model system to investigate floral organs development under different photoperiods. Morphological changes of intermediate cleistogamous (inCL) flowers have been observed, the trends in variation of changes from CH flowers to CL flowers or from CL flowers to CH flowers have been analyzed, the localized effects of poorly developed stamens and petals in CL and inCL flowers have been identified. This research provided morphology and structural changes with implication for the evolutionary significance of the dimorphic flower formation for further study in dimorphic flower development.Methods We used methods of anatomy and structural analysis to observe the morphological structures of flowers under different photoperiods.Important findings Photoperiod played an important role in the development of CH and CL flowers in V. philippica. Under short-day light and intermediate-day light, both CH and inCL flowers developed simultaneously. Most of the floral buds were CH flowers under a photoperiod of short-day light, but most of the floral buds were inCL flowers under mid-day light. Complete CL flowers formed under long-day lights. However, there were a series of transitional types in the number and morphology of stamens and petals among inCL flowers, including five stamens with three petals related to CH flowers and two stamens with one petal related to CL flowers. The former type was dominant under short-day light conditions, and the latter type was dominant under mid-day light. Further more, there were localized effects in stamen and petal development for CL and inCL flowers. The development of ventral lower petal (corresponding to the lower petal with spur of CH flower) and the adjacent two stamens in inCL flowers were best, and the back petal was similar to that of CL flowers, an organ primordium structure. The adjacent stamens with the back petals tended to be poorly developed. In extreme cases, these stamens in inCL flowers had no pollen sac, only a membranous appendage or even a primordium structure. When the plants with CL or CH flowers were placed under short-day light or long-day light, the newly induced flowers all showed a series of inCL flower types, finally the CL flowers transformed into CH flowers, and the CH flowers transformed into CL flowers. This result indicates the gradual effects of different photoperiods on dimorphic flowers development of V. philippica. A long photoperiod could inhibit the development of partial stamens and petals, and a short photoperiod could prevent the suppression of long-day light and promote the development of stamens and petals.     

地丁化学成分的研究

从紫花地丁中首次分得四个有机酸,一个酰胺和一个黄酮甙,其中二十四酰对羟基苯乙胺为一新化合物。     

紫花地丁春花与夏花的发育条件

紫花地丁的两种花紫花地丁(Viola spp.)是遍生我国原野上的草花,我国旧文献中已有记载:此花"处处有之,其叶似柳而微细,夏开紫花,结角……"(本草纲目),又有宿根,"根直     

宝山堇菜(Viola baoshanensis)、紫花地丁(V. yedoensis)光合异质性比较

利用荧光成像技术,研究了特定和不同光合有效辐射下,宝山堇菜(Viola baoshanensis)、紫花地丁(V.yedoensis)不同叶龄(幼叶和成熟叶片)叶片纵向(叶基、叶中部、叶尖)间的光合异质性特征。特定光化光照射下,两种堇菜不同叶龄的Fv/Fm、ΦpsⅡ、qP、PS/50和Abs在叶尖、叶中部、叶基间呈依次降低趋势,NPQ/4和qN变化趋势与之相反。Fv/Fm、ΦpsⅡ和Abs在两种堇菜不同叶龄的叶片纵向间均没有显著性差异,NPQ/4和qN均显示宝山堇菜不同叶龄的叶片纵向间存在显著差异;但NPQ/4和qN分别显示紫花地丁成熟叶和幼叶叶片的叶尖和叶基处差异显著。qP的显著差异只存在于宝山堇菜幼叶的叶尖和叶基处,PS/50在两种植物幼叶纵向间均有显著差异。快速光曲线的变化中,两种堇菜α由叶尖向叶基下降幅度不明显,幼叶纵向间Pm差异显著,成熟叶叶尖处Pm显著高于叶中部和叶基。两种堇菜成熟叶叶尖处Ik显著高于叶中部和叶基,宝山堇菜幼叶纵向间Ik差异显著,而紫花地丁幼叶纵向间Ik差异不显著。以上结果反映出两种堇菜叶片纵向间Fv/Fm和ΦpsⅡ具有较高的均质性,Fv/Fm和ΦpsⅡ的下降受到NPQ/4、qN、qP、PS/50和Abs的综合影响,但Fv/Fm和ΦpsⅡ的变化与NPQ/4、qN、qP、PS/50的显著变化并不一致。叶龄对两种堇菜叶片纵向间α影响不显著,对Pm影响显著,α的小幅下降反映两种堇菜叶片纵向间捕光能力基本相同。叶片纵向间Ik的显著差异受叶龄和植物种类的综合影响。     

宝山堇菜（Viola baoshanensis）镉胁迫下的光合作用

利用荧光成像技术研究了自然生长状态下宝山堇菜、紫花地丁镉含量与光合作用间的关系。野外调查表明,宝山堇菜、紫花地丁镉含量差异显著,地上部分镉含量分别为867mg/kg和2mg/kg,相差近450倍;镉转运系数、富集系数,宝山堇菜均大于1,紫花地丁均小于1。叶绿素荧光参数,宝山堇菜F_m、I_k、P_m均显著低于紫花地丁（p=0.05）;两种植物间F_o、F_v/F_m、α均无显著差异。紫花地丁地上部分镉含量与叶绿素荧光参数的相关性较低,宝山堇菜地上部分镉含量与F_m、P_m、I_k达到极显著负相关。F_v/F_m,宝山堇菜、紫花地丁均在0.8左右,与镉的相关性较低,表明其光合作用没有受到镉胁迫。但是,宝山堇菜Fm显著低于紫花地丁,而且与镉呈负相关,表明宝山堇菜叶片Fm会随镉含量的增加而降低,预示镉可能会造成宝山堇菜光系统Ⅱ叶绿素结构破坏或者抑制其电子传递。参数I_k和P_m在两种植物间差异显著及其与宝山堇菜地上部分镉含量显著负相关反映宝山堇菜光耐受能力和电子传递能力在镉胁迫下逐渐减弱。初始斜率α始终维持在较高水平,说明镉胁迫并未对宝山堇菜捕光系统造成显著伤害。结合Fm值的变化和快速光曲线中荧光参数的变化推断镉最有可能会造成宝山堇菜电子传递能力的减弱。     

紫花地丁对盐胁迫的生理反应

为研究盐分胁迫下紫花地丁生理反应,本文以紫花地丁幼苗为试验材料,对不同浓度NaCl处理中的幼苗叶片,进行光合色素、蛋白质、可溶性糖、游离脯氨酸的测定。结果表明,盐胁迫下紫花地丁叶片光合色素含量降低、蛋白质、可溶性糖、游离脯氨酸含量呈先升高后下降的趋势,当处理浓度高于NaCl2%时,各指标与对照之间差异显著。紫花地丁幼苗能够正常生长的抗盐阈值为NaCl2%,可耐低盐胁迫。渗透调节物质是紫花地丁的主要耐盐性指标。     

宝山堇菜（Viola baoshanensis）、紫花地丁(V. yedoensis)光合异质性比较

利用荧光成像技术,研究了特定和不同光合有效辐射下,宝山堇菜（Viola baoshanensis）、紫花地丁 (V. yedoensis)不同叶龄（幼叶和成熟叶片）叶片纵向（叶基、叶中部、叶尖）间的光合异质性特征。特定光化光照射下,两种堇菜不同叶龄的F_v/F_m、ΦpsⅡ、qP、PS/50和Abs在叶尖、叶中部、叶基间呈依次降低趋势,NPQ/4和qN变化趋势与之相反。F_v/F_m、ΦpsⅡ和Abs在两种堇菜不同叶龄的叶片纵向间均没有显著性差异,NPQ/4和qN均显示宝山堇菜不同叶龄的叶片纵向间存在显著差异;但NPQ/4和qN分别显示紫花地丁成熟叶和幼叶叶片的叶尖和叶基处差异显著。qP的显著差异只存在于宝山堇菜幼叶的叶尖和叶基处,PS/50在两种植物幼叶纵向间均有显著差异。快速光曲线的变化中,两种堇菜α由叶尖向叶基下降幅度不明显,幼叶纵向间P_m差异显著,成熟叶叶尖处P_m显著高于叶中部和叶基。两种堇菜成熟叶叶尖处I_k显著高于叶中部和叶基,宝山堇菜幼叶纵向间I_k差异显著,而紫花地丁幼叶纵向间I_k差异不显著。以上结果反映出两种堇菜叶片纵向间F_v/F_m和ΦpsⅡ具有较高的均质性,F_v/F_m和ΦpsⅡ的下降受到NPQ/4、qN、qP、PS/50和Abs的综合影响,但F_v/F_m和ΦpsⅡ的变化与NPQ/4、qN、qP、PS/50的显著变化并不一致。叶龄对两种堇菜叶片纵向间α影响不显著,对P_m影响显著,α的小幅下降反映两种堇菜叶片纵向间捕光能力基本相同。叶片纵向间I_k的显著差异受叶龄和植物种类的综合影响。     

宝山堇菜(Viola baoshanensis)镉胁迫下的光合作用

利用荧光成像技术研究了自然生长状态下宝山堇菜、紫花地丁镉含量与光合作用间的关系。野外调查表明,宝山堇菜、紫花地丁镉含量差异显著,地上部分镉含量分别为867mg／kg和2mg／kg,相差近450倍;镉转运系数、富集系数,宝山堇菜均大于1,紫花地丁均小于1。叶绿素荧光参数,宝山堇菜Fm、Ik、Pm均显著低于紫花地丁（p=0．05）;两种植物间Fo、Fv/Fm、α均无显著差异。紫花地丁地上部分镉含量与叶绿素荧光参数的相关性较低,宝山堇菜地上部分镉含量与Fm、Pm、Ik达到极显著负相关。Fv／Fm,宝山堇菜、紫花地丁均在0．8左右,与镉的相关性较低,表明其光合作用没有受到镉胁迫。但是,宝山堇菜Fm显著低于紫花地丁,而且与镉呈负相关,表明宝山堇菜叶片Fm会随镉含量的增加而降低,预示镉可能会造成宝山堇菜光系统Ⅱ叶绿素结构破坏或者抑制其电子传递。参数Ik和Pm在两种植物间差异显著及其与宝山堇菜地上部分镉含量显著负相关反映宝山堇菜光耐受能力和电子传递能力在镉胁迫下逐渐减弱。初始斜率α始终维持在较高水平,说明镉胁迫并未对宝山堇菜捕光系统造成显著伤害。结合Fm值的变化和快速光曲线中荧光参数的变化推断镉最有可能会造成宝山堇菜电子传递能力的减弱。     

紫花地丁的组织培养与快速繁殖

1植物名称紫花地丁(Viola philippica) 2材料类别叶片叶柄3培养条件分化与增殖培养基:①MS 6-BA0．5 mg／L;②MS 6一BA1 mg／L;③MS 6-BA1．5 mg／L;④MS 6-BA 2．0 mg／L;⑤MS 6-BA3．0 mg／L[注:MS(大量元素)、BA (6-苄基腺嘌呤,6-benzylaminopurine)NAA(萘乙酸,