Effects of <i>Piriformospora indica</i> on the Respiration of <i>Taxus chinensis</i> var. <i>mairei</i> under Water Stress

Seedlings of Taxus chinensis var. mairei were used as experimental materials to study the adaptation of Piriformospora indica to this plant under water stress. The materials were divided into two groups, namely, with or without inoculation with P. indica. Each group was subjected to four different levels of water stress. Vitality and physiological and biochemical indexes of the roots of T. chinensis var. mairei were regularly measured. Under water stress, T. chinensis var. mairei had significantly decreased root vitality; root vitality was higher in inoculated roots than in uninoculated roots. Under intense water stress, the inoculated roots had a higher soluble sugar content than the uninoculated roots. Under water stress, T. chinensis var. mairei experienced decreased activity of aerobic respiratory metabolic enzymes. The activity of anaerobic respiratory metabolic enzymes and alcohol dehydrogenase initially increased and then decreased, whereas that of lactate dehydrogenase increased. The inoculated roots had a higher activity of respiratory metabolic enzymes than the uninoculated roots. As water stress was further intensified, the roots had significantly decreased activity of aerobic respiratory metabolic enzymes and significantly increased activity of anaerobic respiratory metabolic enzymes. The activity of respiratory metabolic enzymes decreased faster in the uninoculated roots than in the inoculated roots. This study demonstrated that Piriformospora indica plays a positive role in enhancing the antihypoxic ability of T. chinensis var. mairei, thereby alleviating plant damage due to water stress.     

不同遮荫强度下南方红豆杉枝叶紫杉醇产量的季节变化

研究了在一个生长季节内遮荫网不同遮荫强度下对人工种植的南方红豆杉(Taxus chinensis var. mairei)枝叶生物量、紫杉醇含量和产量季节变化。结果表明,在遮荫网89％和46.4％遮光条件下,南方红豆杉枝叶生物量、紫杉醇含量及其产量随发育节律呈现明显的规律性季节变化。在遮荫网89％和46.4％遮光条件下,89％遮荫条件下南方红豆杉枝叶生物量在整个生长季节内都明显高于46.4％遮荫条件下南方红豆杉枝叶生物量;在遮荫网89％和46.4％遮光条件下南方红豆杉枝叶中紫杉醇含量在5月中旬、7月末和11月末都出现较高峰值,后者紫杉醇含量峰值都明显比前者紫杉醇的含量高;89％和46.4％遮荫网遮光条件下南方红豆杉枝叶中紫杉醇的产量都在11月末期时达到最高,后者明显高于前者。因此,生产实践中可采用46.4％遮荫网遮光,采收的最佳季节为11月末期。     

南方红豆杉水浸提液对喜树种子发芽和幼苗生长的化感作用

化感作用是影响人工混交林种间关系的重要因素之一。已有研究表明：喜树南方红豆杉混交对喜树生长有明显促进作用,并从混交改善混交林地微气候角度解释了这种促进作用,但未从种间化感作用角度探讨这种作用。为了探究南方红豆杉是否对喜树具有潜在的化感促进作用,从而更全面了解喜树南方红豆杉混交林种间关系,考察了不同浓度（25、50 g/L和100 g/L）的南方红豆杉（Taxus chinensis var.mairei）鲜叶、凋落叶、枝和根水浸提液对喜树（Camptotheca acuminata）种子发芽和幼苗生长的影响。结果表明：（1）南方红豆杉凋落叶浸提液对喜树种子发芽和幼苗生长无显著影响（P>0.05）,而鲜叶、枝和根浸提液对喜树发芽和幼苗生长均具有不同程度的促进作用（P<0.05）,且作用强度大体与浓度呈正相关;（2）100 g/L的南方红豆杉鲜叶、枝和根浸提液浸泡的喜树种子,其发芽率比对照组分别提高8.1%、14.9%和25.6%;（3）南方红豆杉鲜叶浸提液只有在高浓度（100 g/L）时,对喜树幼苗基径生长具有促进作用（P<0.05）,而对其株高、全株干重和净光合速率无显著影响;南方红豆杉根和枝浸提液对喜树幼苗株高、基茎、干重和净光合速率均具有促进作用（P<0.05）,与对照组相比,100 g/L的根和枝浸提液浇灌喜树幼苗,可以使喜树幼苗株高分别提高14.2%和8.4%,基径分别提高19.0%和15.3%,干重分别提高23.1%和15.9%,净光合速率分别提高11.6%和6.1%。研究结果表明,在喜树南方红豆杉混交林中,南方红豆杉对喜树的正向化感作用可能是促进喜树生长的重要因素之一。     

Inhibitory effect of 13 taxane diterpenoids from Chinese yew (Taxus chinensis var. mairei) on the proliferation of HeLa cervical cancer cells

The inhibitory effect of 13 taxanes isolated from the Chinese yew (Taxus chinensis var. mairei) on the proliferation of human cervical cancer HeLa cells were examined using an MTT assay. Four compounds having a hydrophobic cinnamate side chain showed antiproliferative activity, which may be due to increased cell permeability.     

南方红豆杉生长发育及其紫杉醇含量与环境因子的关系

研究了人工种植的3年生南方红豆杉的生长发育特征及其枝叶中紫杉醇的含量,并观测了其生长环境因子的动态特征。通过对南方红豆杉的生长发育特征以及其枝叶中紫杉醇含量与生长环境因子进行相关分析发现,南方红豆杉生长发育与空气和土壤湿度呈极显著的正相关,与空气和土壤温度、土壤N和K含量、土壤有机质和土壤紧实度呈正相关,与净辐射和土壤P含量呈负相关,但相关关系不显著,说明高温、高湿以及遮荫有利于南方红豆杉的生长,且对土壤P含量有较高需求;南方红豆杉枝叶中紫杉醇含量与空气和土壤湿度、土壤N、P和K含量呈负相关,与净辐射、空气和土壤温度、土壤有机质和土壤紧实度呈正相关,但相关关系均不显著,说明高温、高光强和干旱促进南方红豆杉枝叶中紫杉醇含量的增加,且紫杉醇含量的增加对土壤中N、P和K含量有一定的需求,尤其是N和K,对P需求不大;南方红豆杉生长发育与其枝叶中紫杉醇含量呈负相关,相关关系也不显著,推测紫杉醇可能是南方红豆杉的防御物质,以促使南方红豆杉抵御高温、干旱和高光强等不良环境。     

南方红豆杉毛状根诱导体系的建立及毛状根中 紫杉醇的分离纯化研究

目的:研究发根农杆菌对南方红豆杉转化的转化体系和毛状根中紫杉醇的提取纯化工艺;方法:采用发根农杆菌对南方红豆杉不同外植体进行诱导,并对影响毛状根转化的因素进行分析。采用超临界CO2流体萃取法、硅胶柱色谱法以及高效液相法对毛状根中的紫杉醇进行分离提纯;结果:成功构建了南方红豆杉毛状根诱导体系。其中农杆菌的种类、外植体类型、预培养和共培养时间、培养基中激素浓度均影响毛状根的转化率。毛状根经高压纸电泳检测显示表达冠瘿碱。毛状根中紫杉醇经超临界CO2法提取及硅胶柱色谱法分离提纯,采用高效液相法分析显示其纯度可达到98%。结论:发根农杆菌诱导南方红豆杉产生的毛状根中可产生紫杉醇,可作为紫杉醇的主要来源。     

遮荫对南方红豆杉光合特性及生活史型影响

对浙江省富阳市种植的5年生南方红豆杉,89%遮荫的条件下生长的南方红豆杉与46.4%遮荫及自然光条件诱导50天的南方红豆杉的光合特性、光合色素及其生活史型的研究。结果表明:89%遮荫、46.4%遮荫和自然光的光补偿点分别为18.88,30.52和65.34 μmol·m^-2·s^-1,光饱和点分别为287.01,258.25和358.92 μmol·m^-2·s^-1,遮荫可以降低南方红豆杉的光补偿点和光饱和点,从而能够更好地利用弱光,同时提高光合速率,增强了南方红豆杉的光合能力,其中以89%遮荫的变化最明显。随着遮荫程度的增大,南方红豆杉叶片的叶绿素a、叶绿素b及类胡萝卜素等光合色素的含量均增大,叶绿素a与叶绿素b含量的比值减小,说明遮荫可以增加植物对光能的利用,尤其是增加了对蓝紫光的利用,提高了光合效能。研究还发现遮荫对南方红豆杉的生活史型有一定的影响,89%遮荫、46.4%和自然光的生活史型分别为V_0.836C_0.164,V_0.625C_0.375和V_0.772C_0.228,以89%遮荫的营养生长最为旺盛。因此,89%遮荫是这三种光照条件中南方红豆杉营养生长的最适条件。     

喜树与南方红豆杉混交群落生活史型谱特征

研究了不同生境条件下喜树和南方红豆杉混交群落中喜树和南方红豆杉的生活史型谱特征。结果表明,喜树和南方红豆杉在S生活史型得分和比例均为0。喜树和南方红豆杉生活史型V主成分得分均以山地2最高,而纯林的生活史型V主成分得分最低,C也表现为山地2最高,而纯林最低。喜树种群的生活史型比例按照从山地、坡地、平地、纯林排序,V型比例逐渐上升而C型比例逐渐下降的趋势,这就构成了混交群落中喜树种群的生活史型谱。南方红豆杉种群的生活史型谱特征,主要表现为山地2和坡地V型比例最小,而C型比例最大,纯林种植中V型比例较高,而以纯林1的V型比例最高,C型比例最小。     

南方红豆杉无性系种群结构和动态研究

研究了元宝山南方红豆杉克隆种群的结构和动态．结果表明,种群S1～S6级个体均存在,S1、S2级个体占总个体数的64％．从元宝山南方红豆杉种群的大小级结构图看,结构呈较规则的金字塔形;南方红豆杉无性系幼树幼苗的贮备较为丰富,种群的年龄结构呈较规则的金字塔形,存活曲线近于倒J形;种群大小级动态指数Vpi=38．26％,考虑外部干扰时,Vpi′：6．38％>0,表现为稳定型种群．南方红豆杉种群更新的主要途径是无性繁殖,种群的数量动态主要受克隆内部调节机制以及外部环境的影响．生存分析结果表明,种群幼树幼苗的生存状态不稳定,死亡的可能性很大,而成年个体的生存状态则较为稳定．     

溶磷细菌和丛枝菌根真菌接种对南方红豆杉生长及根际微生物和土壤酶活性的影响

以南方红豆杉实生苗为材料,采用盆栽实验探讨了高效溶磷细菌草木樨中华根瘤菌(Sinorhizobium meliloti)CHW10B与丛枝菌根真菌缩球囊霉(Glomus constrictum)单独和双接种条件下,其植株生长、根际土壤可培养微生物数量、土壤酶活和土壤微生物功能多样性变化,在微生态水平揭示接种对南方红豆杉生长的影响及其机制。结果表明:(1)各接种处理对南方红豆杉幼苗均有促生长作用,接种处理苗高、地径和生物量均较对照显著增加,并以双接种促进效果最好。(2)各接种处理提高了南方红豆杉根际土壤可培养细菌、真菌和放线菌含量,增加了土壤微生物碳源利用率,改变了土壤中物种的丰富度和均一度,增加了土壤中的生物多样性。(3)各接种处理促进了南方红豆杉根际重要土壤酶(酸性磷酸酶、脱氢酶、转化酶)活力的增加,且双接种的促进作用最为明显。可见,溶磷细菌(草木樨中华根瘤菌CHW10B)和丛枝菌根真菌(缩球囊霉)具有协同作用,两者同时接种可显著提高南方红豆杉根际土壤微生物数量及土壤酶活力,提高土壤微生物碳源利用率和土壤肥力,增加土壤中的生物多样性,从而达到间接促进宿主植物南方红豆杉生长的目的。     

南方红豆杉丛枝菌根(AM)的研究

研究了南方红豆杉根部丛枝菌根真菌(AMF)侵染情况、菌根形态结构以及根际土中AMF孢子的种类与数量.结果显示:南方红豆杉可与AMF形成典型的丛枝-泡囊型菌根,侵染率在71.2%~94.4%,但是历山、蟒河自然保护区的侵染强度优于人工栽培区;在南方红豆杉根际土中共分离鉴定出5种AMF,无梗囊霉属1种、球囊霉属4种,分别是:光壁无梗囊霉、地表球囊霉、地球囊霉、缩球囊霉、明球囊霉,其中光壁无梗囊霉为优势种;南方红豆杉的根由表皮、外皮层、内皮层、中柱组成,AMF只侵染表皮层、内皮层,不能侵染中柱.这为将来利用AMF接种技术进行南方红豆杉的繁殖、移植栽培以及紫杉醇的积累等研究提供了理论依据.     

光胁迫对南方红豆杉叶片中叶绿体色素和紫杉醇含量的影响

研究了在光胁迫下南方红豆杉叶片中叶绿体色素含量和紫杉醇含量的动态变化。结果表明,与对照相比,叶绿素a、叶绿素b和总叶绿素的含量以及叶绿素a/叶绿素b比值在光胁迫下均减少,随胁迫时间的延长,叶绿素a、叶绿素b和总叶绿素的含量逐渐上升,叶绿素a与叶绿素b的比值先上升后下降;类胡萝卜素含量在前两周与对照相比降低,2周后高于对照;类胡萝卜素与叶绿素的比值高于对照,随胁迫时间的延长,其趋势先降低后上升。光胁迫处理后,南方红豆杉叶片中紫杉醇含量与对照相比,在第1周时略有下降,而后随胁迫时间的延长,紫杉醇含量在第2周开始迅速大量积累,而在处理3周时达到较高值,已增加到1.5倍,这对提高人工种植的南方红豆杉中紫杉醇含量具有非常重要的意义。     

Mycorrhizal Fungal Effects on Growth,Antioxidant Capacity,and Medicine Quality of <i>Paris polyphylla</i> var. <i>yunnanensis</i>

A field experiment was conducted to determine the effects of two commercial strains composed of mulple arbuscular mycorrhizal fungi (AMF) species on plant growth, antioxidant capacity, and medicine quality of Paris polyphylla var. yunnanensis in three subtropical soils from Wanzhou, Anshun and Baoshan in fields. The results showed that AMF inoculation enhanced the fungal colonization rate and activities of both succinate dehydrogenase and alkaline phosphatase, thereby, enhancing the mycorrhizal viability of P. polyphylla var. yunnanensis. The concentrations of photosynthetic pigments (chlorophyll a, b, a+b and carotenoids), soluble sugar, soluble protein and photosynthetic capacity were higher in AMF-inoculated plants than in non-AMF-treated plants in field. AMFtreated plants recorded higher activities of catalase, peroxidase and superoxide dismutase, and caused the reduction in malondialdehyde content, indicating lower oxidative damage, compared with non-AMF plants. Polyphyllin I, Polyphyllin II, Polyphyllin III, Polyphyllin IV and total polyphyllin contents were increased by AMF treatment. In conclusion, AMF improved the plant growth, antioxidant capacity and medicinal quality of P. polyphylla var. yunnanensis seedlings. Hereinto, AMF effects on the soil from Wanzhou was relatively greater than on other soils.     

濒危植物南方红豆杉种群生命表及谱分析

Based on the investigation in Longxi Mountain National Nature Reserve and the theory of survival analysis, a static life table of Taxus chinensis var. mairei population was worked out, the curves of its survival rate, mortality rate and killing power were drawn, and the population dynamics was analyzed by spectral analysis. The results showed that the survival curve of the population appeared to be a type of Deevey-Ⅲ, and the high mortality of seeding was one of the important reasons which caused Taxus chinensis var. mairei to be endangered. The spectral analysis of the population showed that there was a marked periodic regularity in the process of natural regeneration of Taxus chinensis var. mairei.     

南方红豆杉盆栽扦插育苗技术研究

南方红豆杉(Taxus chinensis var. mairei)是中国特有的国家一级保护植物。2009-2010年采用2年龄实生苗为采条母株进行室内盆栽扦插试验研究,结果表明:珍珠岩和河沙两种基质相比较,珍珠岩上生根率显著高于河沙,是更佳选择;用市售的"花语牌"快速促根剂配制成5、10、15、2 0mg/mL 4个浓度分别处理插穗基部并以蒸馏水作对照发现,10 mg/mL、15 mg/mL处理有利于生长,15 mg/mL处理后生根率显著高于5 mg/mL、10 mg/mL、20 mg/mL及对照。再以同一浓度生根粉分别浸泡4 h、8 h、10 h作对比发现,8 h和10 h处理后生长效果较好,10 h的生根率显著高于4 h、8 h。     

南方红豆杉根际溶无机磷细菌的筛选、鉴定及其促生效果

【目的】对南方红豆杉(Taxus chinensis var.mairei)根际溶无机磷细菌进行了分离、筛选与鉴定,并对获得的高效溶磷菌株进行了温室盆栽试验。本研究为通过生物途径改善南方红豆杉磷素供应,促进其生长提供了优良的菌株资源。【方法】利用选择培养基从南方红豆杉根际土壤中共分离出具溶磷能力的细菌;采用NBRI-BPB培养基进行复筛获得溶磷能力较强的溶无机磷细菌;并采用钼锑抗比色法测量其在NBRIP培养基中经4d发酵后的可溶性磷含量;通过形态指标、生理生化测定、Biolog系统和16S rDNA序列分析鉴定细菌种类;并进行了溶磷菌株的室内盆栽实生苗接种试验。【结论】从南方红豆杉根际共分离出4株高效溶磷细菌,分别鉴定为荧光假单胞菌(Pseudomonas fluorescens)、蜡状芽胞杆菌(Bacillus cereus)、草木樨中华根瘤菌(Sinorhizobium meliloti)和地衣芽胞杆菌(Bacillus licheniformis);4株细菌对南方红豆杉苗期的生长有明显的促进作用。     

Microsatellite loci from Taxus chinensis var. mairei (Taxaceae), an endangered and economically important tree species in China

Taxus chinensis var.mairei is an endangered and economically important tree species in China.Four polymorphic microsatellite markers were developed and characterized using the fast isolation by amplified fragment length polymorphism (AFLP) of sequences containing repeats (FIASCO) protocol.Polymorphism of each locus was quantified for five natural populations from Hubei Province,China.The number of alleles per locus varied from 6 to 24.The ranges of observed and expected heterozygosity were 0.493-1.000 and 0.440-0.845,respectively.These are the first microsatellites reported for the family of Taxaceae and will be helpful for the ongoing population and conservation genetics research of Taxus chinensis var.mairei.     

Structural and Histochemical Features of the Slow-Growing Perennial <i>Coptis chinensis</i> Franch. (Ranunculaceae)

Huanglian (Coptis chinensis Franch.) is a slow-growing perennial medicinal herb with considerable economic value. This study aimed to determine the structural characteristics and the levels of berberine deposits in the organs and tissues of Huanglian using light and epifluorescence microscopy. The adventitious roots are composed of primary and secondary structures with endodermis, exodermis, and phellem. The rhizome structures are composed of primary and secondary structures with cuticle and phellem. The leaves are composed of sclerenchymatous rings, isolateral mesophyll, and thin cuticles. We detected berberine in the xylem walls of the roots and rhizomes as well as in the sclerenchymatous rings of the petioles. We postulate that as the exodermis is developed, the deposition of berberine in the xylem closest to the root tips may affect water and nutrient absorption and transfer. Leaf blades had a thin cuticle and isolateral mesophyll, suggesting shade tolerance. These structural and histochemical features suggest that Huanglian is adapted to the slow growing nature of a shady environment.     

不同栽培条件下南方红豆杉生长特性研究

以4～6年生南方红豆杉为材料,对其不同栽培条件下地径、冠幅、株高、当年抽高、当年侧枝数及总侧枝数等生长特性进行了研究。结果表明:南方红豆杉4～6年生幼林生长量随年龄增大而增加,但其增加的趋势变缓,生长速度变慢;耕作农田中南方红豆杉生长最好,土壤板结未耕作农田生长较差,去除上层20cm耕作土的农田最差;南方红豆杉纯林生长好于落叶阔叶林下(林分郁闭度0.6～0.7),且种植密度越大差异越明显;南方红豆杉落叶阔叶林下套种及纯林生长量均随栽培密度减小而增大,不同栽培密度相差越大,生长量差异越明显。