珍稀濒危药用植物黄檗野生种群遗传多样性的AFLP分析

黄檗(Phellodendronamurense)是我国珍贵的药用和用材树种,现已被列为国家二级保护植物。为了研究其遗传多样性和遗传结构,以制定有针对性的保护策略,作者采用8对荧光引物,对来自10个野生种群共计129个黄檗个体进行了AFLP分析。共扩增出1,704条谱带,其中多态带1,581条。遗传分化系数Gst为0.3420,即有34.20%的遗传变异来自种群间。黄檗物种水平的遗传多样性高于种群水平:在物种水平上,多态位点百分率PPL=92.77%,有效等位基因数Ne=1.4636,Nei’s基因多样性指数H=0.2316,Shannon信息指数I=0.4275;在种群水平上,PPL=54.20%,Ne=1.2487,H=0.1524,I=0.2371。UPGMA聚类结果表明,黄檗野生种群可分为四类,并且来自同一地区的黄檗个体有聚为一类的趋势,说明其遗传变异可能与生境有关。建议在各种群内大量采样,实施迁地保护;在长白山、完达山、大兴安岭、小兴安岭、千山以及燕山山脉的云蒙山等地区,均匀布点划分区域进行就地保护,保护种群完整性,促进种群自我更新。     

燕山山脉黄檗种群结构与动态特征

黄檗（Phellodendron amurense）为我国珍稀濒危物种，具有较高的药用价值。以燕山地区野生黄檗种群为研究对象，采用典型抽样对8个保护区内的黄檗进行全面调查，通过编制静态生命表并绘制存活曲线，分析种群年龄结构和种群变化规律，并通过种群动态量化和时间序列预测分析种群未来数量变化。结果表明：（1）黄檗种群结构总体上呈倒"J"型，但存在外部干扰时，种群结构数量变化动态指数为1.017%，表明该种群趋于稳定型；（2）静态生命表及相关曲线分析显示，黄檗种群存活数量整体上随着径级增大，个体数量逐渐下降；黄檗种群的生存期望在不同径级间存在波动，而第Ⅶ径级的生存期望最高，种群存活曲线符合Deevey-Ⅱ型，黄檗种群具有前期减少、中期稳定、后期衰退的特点；（3）时间序列预测分析表明，在未来2、4、6、8个径级之后，黄檗中老龄个体数量将逐渐增加。黄檗种群濒危的主要因素是幼苗数量少，且呈现散生分布状态，建议加强幼苗抚育工作，提高幼苗的存活率，保护和改善生境，从而促进黄檗种群的自然更新和恢复。     

塔里木河上游胡杨种群结构与动态

采用样带调查法对塔里木河上游3个断面8.5 hm2天然胡杨林进行野外调查,绘制了胡杨种群年龄结构图和存活曲线,结合种群动态量化方法与时间序列预测,分析种群结构与动态特征。结果表明:塔里木河上游胡杨种群密度随径级增大而减小,自疏指数接近-3/2,个体胸径达24 cm后种群密度保持相对稳定。中、下段胡杨种群年龄结构呈金字塔型,幼龄级所占比例较大、林分年轻;存活曲线为Deevey C型,且动态指数0,为增长种群。受所处生境条件影响,上段不同生境胡杨种群年龄结构差异明显。阿瓦提县胡杨种群结构呈金字塔型,幼龄级比例高达77.2%,存活曲线属Deevey C型,为增长种群;南口镇与托海乡胡杨种群结构呈壶型,幼龄级比例极低、中大龄级比例高达84.7%,存活曲线属Deevey A型,为衰退种群;沙黑里克镇胡杨种群结构呈钟型,存活曲线属Deevey A型,为暂时稳定种群。种群数量动态与时间序列分析表明,中、下段胡杨更新资源丰富,未来种群能够稳定增长;上段南口镇、托海乡与沙黑里克镇幼龄级数量随时间推移减少,种群向大老龄级发展,未来种群将走向衰退。总之,上段胡杨种群因更新资源不足而趋于衰退,未来保持适当的人工辅助恢复措施和加强保护现有植株及其生境是保持胡杨种群自然更新和进行种群恢复的关键。     

濒危植物太白红杉种群年龄结构及其时间序列预测分析

为了阐明太白红杉(Larix potaninii var.chinensis)种群的年龄结构和未来发展趋势,合理保护现有资源,在太白山地区调查了 29个样地(10 m×10m),对处于不同生境的 5个种群的年龄结构、静态生命表和时间序列预测进行了分析。种群年龄结构分析表明,各种群个体数量主要集中于Ⅲ、Ⅳ、Ⅴ、Ⅵ、Ⅶ龄级(个体数量比重占68.64％),幼、老龄个体数量较少。受所处生境条件影响,不同种群年龄结构特点不同:海拔较低的种群(B)由于水热条件适宜,林内有林窗出现,幼龄级个体数相对丰富;而其他种群由于幼龄个体严重缺乏,衰退趋势明显。太白红杉不同种群生命表和存活曲线的分析表明,尽管所处的生境差异较大,但存活曲线基本属于Deevey Ⅲ型,种群偏离典型存活曲线的程度与幼苗缺乏程度有关,一般V-Ⅸ龄级死亡率较高。时间序列分析表明,在未来20、60、100和200年中,各太白红杉种群均会呈现老龄级株数先增加后减少的趋势,种群稳定性维持困难。太白红杉种群年龄结构和动态趋势与银杉(Cathaya argyrophylla)等其他濒危植物比较,导致种群衰退的原因相似:在郁闭的林下种子萌芽和幼苗生长困难。应该利用太白红杉喜光、耐旱和中老年个体产种量丰富的特性,对现有太白红杉林分实行就地保护,合理抚育管理。低海拔地区,应适当间伐非目的树种     

新疆濒危野生樱桃李的种群结构与动态

为了解野生樱桃李种群结构和种群生态现状,采用“空间序列代替时间变化”的方法,对新疆天然分布区内的野生樱桃李不同种群的径级结构、冠幅结构及株高结构进行了分析,运用静态生命表描述了野生樱桃李种群年龄结构的动态变化规律.结果表明,樱桃李天然种群的株高和冠幅结构呈现为一定的衰退趋势,冠幅在总体呈现为近乎对数型的衰减趋势;种群的径级结构趋于稳定型,但种群结构表现出较强的波动性,种群衰退风险较高;同时,不同样地种群因所处微环境等的差异,在径级、株高和冠幅结构上表现出一定差异.野生樱桃李种群的存活曲线接近于B1型(台阶型),说明幼龄的死亡率较高,且不同生活史时期的死亡率差异较大.新疆的野生樱桃李天然种群表现出一定的不稳定性,亟待补充低龄级的个体,并减少对成年树的砍伐破坏,从而为种群的保育和恢复提供保障.     

长白山水曲柳种群动态

通过对长白山水曲柳种群的调查,以种群径级结构代替年龄结构,采用空间代替时间的方法,对水曲柳种群的年龄结构、静态生命表以及种群时间动态预测进行了分析。结果显示:长白山水曲柳种群幼龄个体偏少,中老龄个体较多。其中阔叶红松林中水曲柳种群主要依靠中老龄植株维持,幼龄个体缺乏;白桦次生林中水曲柳植株龄级较阔叶红松林幼龄,但仍然存在幼树缺乏现象。长白山水曲柳种群的存活曲线基本属于偏正态DeeveyⅢ型,总体呈现衰退趋势。种群时间动态预测显示,随着水曲柳老龄个体短暂增多后的逐渐减少,水曲柳种群逐渐衰退。因此建议在立足现有种群的维持保护上,研究水曲柳濒危机制,采取适当人工抚育措施,保证种群持续稳定发展。     

濒危植物秦岭冷杉种群数量动态

为了对濒危植物秦岭冷杉种群数量动态评价和预测,通过样地调查和数据统计,研究了秦岭冷杉种群的年龄结构、静态生命表及其与环境因子关系,运用时间序列模型预测了种群数量动态.结果表明,多数秦岭冷杉种群幼龄级个体数较少,中老龄个体数量较大,呈衰退趋势.仅处于低海拔地区的秦岭冷杉-木蓝-苔草群丛中的种群(D种群)由于立地条件较好,幼龄级个体数量相对丰富,种群稳定.不同秦岭冷杉种群生命表和存活曲线的分析表明,尽管生境条件差异,但存活曲线基本接近DeeveyⅢ型;不同种群偏离典型存活曲线的程度与幼苗缺乏程度有关,一般Ⅲ～Ⅴ龄级死亡率较高.时间序列分析表明,在未来20、40和80年中,不同秦岭冷杉种群均会呈现老龄级株数先增后减的趋势,种群稳定性长期维持困难.对影响秦岭冷杉种群增长的10个环境因子通过主成分分析(PCA)发现,乔木层盖度、土壤有机质含量和空气湿度对种群发挥有利影响,而人为干扰和光照强度对秦岭冷杉种群增长发挥不利影响.应充分利用秦岭冷杉性喜荫、耐寒、种子活力较强的特点,加强现有林分就地保护,重点是具有结实能力的中老龄个体;在阴坡地带,对林下灌木比较密集的群丛,通过砍灌、清理林下活地被物等抚育措施,为幼苗发育创造良好的环境条件;就地采种育苗,扩大人工种群.     

不同植被地区天然黄檗生物碱含量对季节的响应

黄檗(Phellodendron amurense Rupr.)不仅为中国特有的濒危植物,还是我国著名中药材"关黄柏"的药源植物,由于其珍贵的药用价值而被过度开采。本研究采用HPLC检测不同植被地区天然黄檗种群中药根碱、掌叶防己碱和小檗碱等生物碱的含量,分析不同部位,不同季节对生物碱分布的影响。研究表明,药根碱在黄檗各部位的分布为:根皮茎皮多年生枝一年生枝叶片。掌叶防己碱含量分布为:茎皮多年生枝皮根皮一年生枝叶片。小檗碱在不同部位的分布规律表现为:根皮茎皮多年生枝一年生枝叶片。根皮和茎皮中的药根碱含量随着春、夏、秋季节的推移呈逐渐上升的趋势,表现为秋季夏季春季,而多年生枝皮、一年生枝、叶片中的药根碱则是秋季春季夏季。各部位的掌叶防己碱均随着季节推移而升高,表现为秋季夏季春季。根皮、茎皮和多年生枝皮中小檗碱含量随着季节变化表现为夏季秋季春季。一年生枝中小檗碱在夏季降低,而到了秋季再升高。     

水分胁迫对黄檗幼苗三种生物碱含量的影响

黄檗（Phellodendron amurense）是东北阔叶红松林的重要伴生树种,也是我国名贵中药关黄柏的药源植物,小檗碱（berberine）、药根碱（jatrorrhizine）、掌叶防己碱（palmatine）是研究者通常关注的主要黄檗生物碱。通过池栽法和渗灌控水方式设置了对照、轻度干旱、重度干旱和水涝（土壤水势分别控制在-40～-20KPa、-80～-60KPa、〈-80KPa和-20～0KPa）4种水分处理。比较了不同水分状况下黄檗幼苗小檗碱、药根碱及掌叶防己碱含量的变化。对于当年生的黄檗幼苗,总体上轻度干旱有利于上述3种生物碱的合成与积累,重度干旱处理下幼苗3种生物碱的含量与对照差异不大。而水涝处理则导致幼苗3种生物碱的含量显著降低。同时,干旱或水涝处理都导致黄檗幼苗生长受抑,幼苗的株高、茎径和生物量都显著低于对照幼苗,因此3种生物碱的单株产量仍是对照的黄檗幼苗最高。茎外皮是中药黄柏的药用部位,也是黄檗幼苗中3种生物碱含号最丰的部位。实验结果预示短期的轻度干旱处理可以提高茎外皮的小檗碱含量。这对以获取生物碱为目的的黄檗幼苗培育可能有参考价值。     

海南吊罗山濒危植物山铜材种群结构与动态特征

为探究海南吊罗山山铜材（Chunia bucklandioides）种群结构与动态特征，以当地山铜材野生种群为研究对象，将种群集中分布区域设为典型样地并开展样地调查，以空间代替时间构建种群年龄结构，采用动态量化分析、绘制存活曲线、计算生存函数等方法分析现有种群结构特点和动态特征，并通过时间序列预测模型推测种群未来发展趋势。结果表明：（1）吊罗山山铜材种群年龄结构接近不规则金字塔型，动态量化分析显示该种群为增长型种群，但第Ⅰ～Ⅲ龄级间存在衰退现象，种群增长性弱，稳定性差，受外界干扰敏感度高，有向稳定型种群转变的趋势。（2）随龄级增加，山铜材标准化存活个体数量减少，生命期望降低，死亡率和消失率总体呈上升趋势，存活曲线趋近于Deevey-Ⅱ型。（3）生存分析显示吊罗山山铜材种群具有前期增长、中期稳定、后期衰退的特点。（4）经过未来2、4、6龄级时间后，Ⅰ～Ⅲ龄级个体数量将逐渐减少，Ⅳ～Ⅶ龄级个体数量少量增加，幼龄苗木自我更新能力不足，种群存在规模收缩和衰退的风险。研究表明，生境碎片化、分布范围小、植株结实量少和幼龄个体不足是导致吊罗山山铜材濒危的主要因素，建议通过设立自然保护小区、辅助种子入...     

High-efficient extraction of principal medicinal components from fresh Phellodendron bark (cortex phellodendri)

There are three key medicinal components (phellodendrine, berberine and palmatine) in the extracts of Phellodendron bark, as one of the fundamental herbs of traditional Chinese medicine. Different extraction methods and solvent combinations were investigated to obtain the optimal technologies for high-efficient extraction of these medicinal components. Results: The results showed that combined solvents have higher extracting effect of phellodendrine, berberine and palmatine than single solvent, and the effect of ultrasonic extraction is distinctly better than those of distillation and soxhlet extraction. Conclusion: The hydrochloric acid/methanol-ultrasonic extraction has the best effect for three medicinal components of fresh Phellodendron bark, providing an extraction yield of 103.12 mg/g berberine, 24.41 mg/g phellodendrine, 1.25 mg/g palmatine.     

光强对黄檗幼苗三种生物碱含量的影响

设置光强分别为全光照的100%(不遮荫)、75%、50%和25%的4种光照处理,比较了不同光强下1年生黄檗幼苗主要药用成分小檗碱、药根碱和掌叶防己碱含量的变化.结果表明,全光照有利于小檗碱、药根碱和掌叶防己碱在茎外皮和根中的合成和积累,总体上随光强减弱,3种生物碱含量降低.相对光强为75%时黄檗幼苗生物量最大,而75%相对光强下黄檗幼苗3种生物碱的单株产量(生物碱含量与生物量之积)最高.     

Effects of exogenous 6-BA and NAA on growth and contents of medicinal ingredient of Phellodendron chinense seedlings

Using Phellodendron chinense seedlings as material, and treated with different concentrations of exogenous 6-Benzylaminopurine (6-BA) and α-naphthyacetic acid (NAA), then observed the growth status. Furthermore, we detected the contents of chlorophyll and soluble sugar, the activities of antioxidases by spectrophotometry, and determined the contents of secondary metabolite by high performance liquid chromatograph. The results showed that different concentrations of exogenous 6-BA increases the fresh weights and plant heights of Phellodendron chinense seedlings, and enhances the contents of chlorophyll and soluble sugar. NAA promoted growth, but deduced the contents of soluble sugar. Compared with control, culturing for 40 d, proper concentrations 6-BA enhanced the activity levels of superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT), proper concentrations NAA increased the activity levels of SOD and CAT, but decreased the levels of POD compared with CK. Suitable concentrations 6-BA enhanced contents of berberine, phellodendrine and palmatine in stems, proper concentrations NAA increased contents of berberine and phellodendrine, but deduced contents of palmatine compared with CK. Based on these results, we concluded that the exogenous 6-BA and NAA had key regulation on the growth and contents of medicinal ingredient of Phellodendron chinense seedlings.     

季节和地理差异对天然黄檗小檗碱和药根碱含量的影响

用高效液相色谱法测定了黄檗根皮、茎皮、多年生枝皮、1年生枝和叶片中小檗碱和药根碱的含量,研究黄檗不同部位两种生物碱的季节和地理差异。结果表明:不同季节,天然黄檗各部位的小檗碱、药根碱含量均表现为:根皮茎皮多年生枝皮1年生枝叶片。小檗碱含量随着季节变化在各部位呈现夏季升高,到秋季下降的趋势。而药根碱在不同部位中的含量随着季节变化的规律不同。根皮、茎皮中的药根碱含量随着季节推移逐渐增多,在夏季升高不明显,而到秋季大量积累。多年生枝皮、1年生枝、叶片中的药根碱在3个季节表现则为先降后升。从纬度水平上看,不同地域间黄檗各部位的生物碱差异显著,在调查的40°—50°N范围内,黄檗两种生物碱的含量普遍在纬度低的地区较高,随着纬度增大先逐渐降低,到N44°左右达到最低值,到了高纬度地区略有回升。     

氮素形态对黄檗幼苗三种生物碱含量的影响

通过改变水培溶液中NH 4-N和NO3--N的比例,探讨了氮素形态对黄檗幼苗主要药用成分小檗碱、药根碱及掌叶防己碱含量的影响。结果表明,小檗碱的含量与氮素形态有明显关系,在根和茎中都表现为NO3--N比例高更有利于小檗碱的积累。单纯的NO3--N供给最有利于药根碱在黄檗幼苗根中的积累,而处理46d后,NH4 -N/NO3--N为75/25和25/75时黄檗幼苗茎中的药根碱含量最高。黄檗幼苗根和茎中的掌叶防己碱含量对营养液中NH4 -N和NO3--N的比例反应类似,在处理46d后,掌叶防己碱含量由高到低所对应的NH4 -N/NO-3-N为25/75、50/50、75/25、0/100、100/0。处理33d后,黄檗幼苗根中的小檗碱、药根碱、掌叶防己碱含量随处理天数的增加呈下降趋势,而在茎中呈上升趋势。如果同时考虑氮素形态对黄檗幼苗生物量的影响,那么NH4 -N/NO3--N为0/100时其小檗碱产量(含量与生物量乘积)最高,而NH4 -N/NO-3-N为25/75时药根碱和掌叶防已碱的产量最高。     

Effects of water stress on berberine, jatrorrhizine and palmatine contents in amur corktree seedlings

Amur corktree (Phellodendron amurense) is one of the important associated species in broadleaved-Korean pine forests, and in the pharmaceutical resource plants of famous Chinese traditional medicine, named cortex phellodendri. Berberine, jatrorrhizine and palmatine are the main alkaloids to which high attention is paid by the researchers. In the present study, water stress treatments with four conditions, that is, mild drought, severe drought, waterlogging, and control (soil water potentials were controlled in the ranges of-40—-20 KPa,-80—-60 KPa, <-80 KPa and-20—0 KPa), were performed using the technique of root-sphere osmotic irrigation in a soil pond. The changes in the main medicinal compositions of berberine, jatrorrhizine and palmatine contents under different water conditions were discussed. As for the annual growing of amur corktree seedlings, mild drought was generally beneficial to the synthesis and accumulation of the three above-mentioned alkaloid contents. The three alkaloid contents did not show great changes under severe drought whereas those contents had significantly reduced under waterlogging compared with controls. Meanwhile, the growth of amur corktree seedlings was inhibited by the treatments of drought and waterlogging. The height, diameter and biomass of amur corktree seedlings were significantly lower than those of the control seedlings, which meant that the three alkaloid contents in a single seedling still kept the highest in control seedlings. Stem cortex was the medical part in Chinese traditional medicine and also kept the most abundant of the three alkaloid contents in amur corktree. Results indicated that short periods of mild drought could improve the berberine contents in the stem cortex, which might have reference value for the cultivation of amur corktree seedlings to obtain alkaloids.     

A range wide geographic pattern of genetic diversity and population structure of Castanea mollissima populations inferred from nuclear and chloroplast microsatellites

Understanding geographical pattern of genetic diversity and population structure is of great importance for formulating conservation and utilization strategies. In this study, we investigated the genetic diversity and population structure of 28 natural populations of Castanea mollissima in China using eight nuclear and six chloroplast microsatellite makers (nSSRs and cpSSRs). Populations from central China harbored the highest genetic diversity at both nSSR and cpSSR markers (nSSR: H _E?=?0.705; cpSSR: H?=?0.461). The standardized measure of genetic differentiation estimated as G′ _ST was 0.447 for nSSR and 0.803 for cpSSR, respectively. The G′_ST-based pollen to seed flow ratio is 3.043, indicating that pollen flow is not extensive among C. mollissima populations. No obvious population genetic structure by geographical locations was found by STRUCTURE analysis based on nSSR data, and similarly, no signal of phylogeographic structure was detected for cpSSR analysis. Five boundaries defining zones of maximum genetic differences within the network of the C. mollissima populations were found, and the locations of those barriers were consistent with those of four mountains, i.e., Daloushan Mountain, Dabashan Mountain, Wushan Mountain, and Qingliangfeng Mountain, indicating that those mountains might act as genetic barriers obstructing the genetic exchange among natural C. mollissima populations. These results provide valuable baseline data for conservation and utilization of this species.     

Population genetic structure of wild boar <Emphasis Type="Italic">Sus scrofa</Emphasis> in Bulgaria as revealed by microsatellite analysis

Over the last two decades wild boar Sus scrofa Linnaeus, 1758 became the most intensively managed game species in Bulgaria. In order to delineate the population genetic structure, which is essential for sustainable wildlife management, we screened 10 porcine microsatellites across 289 wild boar samples originating from all relevant bioregions of the country. Results based on F _ST values, Bayesian clustering methods and a multi-dimensional scaling analysis can be summarised as follows: (1) two main genetic groups were revealed for the Bulgarian data set: the first one included individuals collected from the Balkan Range Mountain and the northern part of the country and the second one comprised individuals from the Rhodope, Osogovo, Iskur Range and Rila mountains in southern and south-western Bulgaria; (2) all Bulgarian wild boar populations showed a higher level of genetic diversity compared to four populations from Germany which were included for outgroup comparison, and (3) wild boar sampled from a game enclosure were found to be genetically divergent from the other Bulgarian populations, indicating human impact on population genetic structure most likely resulted from fencing and former translocation actions. The evolutionary background leading to the two defined management units as well as conservation and management strategies are discussed.     

Effects of domestication on genetic diversity in Chimonanthus praecox: Evidence from chloroplast DNA and amplified fragment length polymorphism data

Chimonanthus praecox (L.) Link is a widely cultivated endemic winter-flowering plant in China that has a long cultivation history. Genetic diversity and genetic structure were compared between wild and cultivated groups to reveal the geographic origin of the cultivated genotypes using chloroplast DNA (cpDNA) sequences and amplified fragment length polymorphism (AFLP) markers. Nine haplotypes were identified using three combined chloroplast fragments. Based on chloroplast data, the wild group showed greater genetic variation and genetic differentiation and a lower measure of gene flow compared to the cultivated group. The AFLP markers also supported this trend. More than 40% of the cpDNA haplotypes were shared between wild and cultivated groups, with shared haplotypes originating from multiple wild populations, suggesting multiple origins of cultivated plants. Moreover, principal coordinate analysis, UPGMA, and structure analysis of AFLP markers revealed that two wild populations clustered with most of the cultivated populations of Ch. praecox, suggesting that most of the cultivated populations mainly originated from these two populations. The combined cpDNA and AFLP results indicated that modern cultivated Ch. praecox experienced multiple events of origin involving two geographic origins, eastern China (Tianmu Mountain) and southwestern China (the border of Hunan–Guangxi–Sichuan–Guizhou).     

Impacts of domestication on population genetics of a traditional Chinese medicinal herb,Atractylodes macrocephala (Asteraceae)

As a medicinal herb, Atractylodes macrocephala Koidz. experienced centuries of cultivation in China, and germplasm resources of all cultivated populations have degraded over time as a consequence of domestication processes. This study used chloroplast DNA and microsatellites to clarify not only the effects of domestication on population genetics, but also determine the geographic origins of landraces. The results revealed that cultivated populations (except the “Pingzhu” landrace) showed higher genetic diversity than their wild counterparts and low levels of genetic differentiation occurred between cultivated and wild groups. Furthermore, STRUCTURE and UPGMA analyses grouped all wild populations into three genotypic clusters, two of which (in Shaanxi and Hunan Province) shared the same gene pool with cultivated A. macrocephala, suggesting that wild populations in Central China have been involved in the origin of cultivated A. macrocephala. Moreover, the wild population from Qimen, Anhui Province and the cultivated “Pingzhu” landrace harbor unique gene pools and rare alleles that could be useful in future breeding efforts. This large‐scale analysis of population genetics on a medicinal herb that has a centuries‐long history of human‐mediated selection will facilitate utilization and conservation of the valuable genetic resources of medicinal species.