Association Analysis of the Growth of Black Poplar (Populus Nigra L.) Under Contrasting Nitrogen Levels

The European black poplar(Populus nigra L.)has been used as a germplasm resource for the breeding of new poplar varieties around the world.The identification and screening of its high nitrogen use efficiency genotypes could enable the breeding of new resource-efficient poplar varieties.The accessions were screened using MALDI-TOF MS genotyping technology for ammonium transporter(AMT)and nitrate transporters(NRT)genes against phenotypic data for seedling height and ground diameter traits,in both low and high nitrogen environments.Allele re-sequencing of seven genes related to root development was carried out using the minisequencing method.By cluster analysis,101 accessions of black poplar were divided into 4 populations,and it was concluded that Central Europe is the origin of the evolution of low-nitrogen and high-efficiency populations of European black poplar.Association study between SNP typing and seedling height and ground diameter traits showed that there were significant correlations between four SNP loci and growth traits under the contrasting N levels.We found that SNP3 and SNP4 in the PttAMT1;3 gene were significantly associated with seedling height traits,and that SNP2 and SNP7 in the PttAMT1;2 and PttAMT1;5 genes,respectively,were significantly associated with ground diameter traits.Thus,considerable allelic diversity is present within the candidate genes studied and can be utilized to develop functional markers to select for poplars with improved growth under N stress conditions.

Pb胁迫对欧美杂交杨(Populus deltoides×Populus nigra)生物量分配格局及其Pb富集特性的影响

采用速生树种修复重金属污染土壤的方法近年来受到越来越多关注,但已有结果存在很大不确定性。为了解杨树在不同Pb胁迫条件下生长响应和Pb富集效果,以长江上游两种典型土壤(酸性紫色土和钙质紫色土)为栽培介质,采用盆栽试验方法,研究了不同Pb浓度处理下(CK:0mg·kg-1;T1:200mg·kg-1;T2:450mg·kg-1;T3:2000mg·kg-1)欧美杂交杨(Populus deltoides×Populusnigra)生物量生产与分配格局以及Pb吸收、富集特性。两种土壤条件下杨树各器官生物量及总生物量均表现出随Pb胁迫程度的增加而降低的趋势,Pb胁迫条件下杨树生物量分配格局在钙质紫色土中表现为茎>粗根>叶>细根。相同浓度Pb处理条件下,单株杨树总生物量均表现为钙质紫色土大于酸性紫色土。随着Pb处理浓度的增大,杨树各器官Pb含量及积累量显著增加。Pb胁迫使杨树对Pb的富集系数逐渐增大而耐性系数逐渐减小。T3处理条件下杨树对Pb的富集系数在酸性紫色土中较大,且各处理条件下杨树对Pb的耐性系数均为酸性紫色土中较大。这些结果表明,高浓度Pb胁迫条件下酸性紫色土中的欧美杂交杨表现出较好的吸收和富集Pb的特性,这为Pb污染土壤的生物修复提供了一定的科学依据。

镉胁迫对欧美杂交杨(Populus deltoides×Populus nigra)光合作用与生物量生产的影响

采用盆栽实验研究了生长在四川盆地典型Cd污染冲积土和紫色土上的欧美杂交杨(Populus deltoides×Populus nigra)对国家土壤环境质量标准中Cd浓度梯度(0、0.3、1.0、2.0 mg/kg)的光合响应和生物生产量。Cd处理显著降低紫色土上生长的杨树叶绿素a和叶绿素总量,增加气孔导度与胞间CO2浓度,增加凋落叶生物量,降低茎生物量、根生物量和单株总生物量。但是,Cd处理对冲积土上生长的杨树叶绿素特征、光合生理指标和生物量生产并无明显差异。尽管同一生长基质上不同Cd处理间杨树光合生理指标与生物量生产均有一定程度的影响,但紫色土上生长的杨树生理和生长特征均显著低于冲积土上生长的杨树。这些结果表明,自然条件国家土壤环境质量标准内,Cd污染并不对杨树生长构成较大威胁,而生长土壤基质的选择可能对杨树的栽培更为重要。这为Cd污染土壤区域城市森林建设和速生丰产林建设提供了一定的科学依据。

Two novel eukaryotic translation initiation factor 5A genes from Populus simonii×P.nigra confer tolerance to abiotic stresses in Saccharomyces cerevisiae

The role of plant eIF5A proteins in multiple biological processes, such as protein synthesis regulation, translation elongation, mRNA turnover, programmed cell death and stress tolerance is well known. Toward using these powerful proteins to increase stress tolerance in agricultural plants, in the present study, we cloned and characterized PsneIFSA2 and PsneIFSA4 from young poplar （P. simonii × P. nigra） leaves. The deduced amino acid sequences of PsneIF5A2 and PsneIF5A4 were 98 % similar to each other, and they are orthologs of eIF5A 1 in Arabidopsis. In a subcellular localization analysis, PsneIF5A2 and PsneIF5A4 proteins were localized in the nucleus and cytoplasm, qRT-PCR analysis showed that PsneIF5A2 and PsneIF5A4 were transcribed in poplar flowers, stem, leaves, and roots. In addition, they were also induced by abiotic stresses. Transgenic yeast expressing PsneIF5A2 and PsneIF5A4 had increased salt, heavy metal, osmotic, oxidative tolerance. Our results suggest that PsneIF5A2 and PsneIF5A4 are excellent candidates for genetic engineering to improve salt and heavy metal tolerance in agricultural plants.

Pharmacological potential of Populus nigra extract as antioxidant,anti-inflammatory,cardiovascular and hepatoprotective agent

Objective:To evaluate antioxidant,anti-inflammatory,hepatoprotective and vasorelaxant activities of Pupulus nigra flower buds ethanolic extract.Methods:Antioxidant and anti-inflammatory activities of the extract were assessed using respectively the ABTS test and the animal model of carrageenan-induced paw edema.Protection from hepatic toxicity caused by aluminum was examined by histopathologic analysis of liver sections.Vasorelaxant effect was estimated in endothelium-intact and-nibbed rings of porcine coronary arteries precontracted with high concentration of U466I9.Results:The results showed a moderate antioxidant activity(40%),but potent anti-inflammatory activity(49.9%)on carrageenan-induced mice paw edema,and also as revealed by histopathologic examination,complete protection against AlCl_3-induced hepatic toxicity.Relaxant effects of the same exlracl on vascular preparation from porcine aorta precontracted with high concentration of U466I9 were considerable at 10^(-1)g/L,and comparable(P>0.05)between endothelium-intact(67.74%,IC_(50)0.04 mg/ml.)and-rubbed(72.72%,IC_(50)=0.075 mg/ml,)aortic rings.Conclusions:The extract exerted significant anti-inflammatory,hepatoprotective and vasorelaxant activities,the latter being cndothelium-independent believed to be mediated mainly by the ability of components present in the extract to exert antioxidant properties,probably related to an inhibition of Ca^(2+)influx.

No consistent daily variation in DNA methylation detected in Populus nigra leaves by methylation-sensitive amplification polymorphism analysis

DNA methylation, an epigenetic mechanism used by cells to control gene expression, has an important biological role in plant development and environmental fitness. Since plant DNA methylation is closely related to environmental conditions, variation during the day is expected. Here, in genetically identical plants of Populus nigra clone N46, DNA methylation changes in leaves over a 24 h period were detected using the methylation-sensitive amplification polymorphism method. The results showed different DNA methylation patterns in mature poplar leaves: not only in individuals at the same time, but also in samples at each of the six time during the day. In addition, night samples had a higher percentage of methylation than in morning samples. However, no statistically significant differences were found among the samples gathered at different times. Similar results were obtained for three other P. nigra clones with different genetic backgrounds. Real time qPCR showed that the DNA methyltransferase genes Pt-MET1 and Pt-SOM1 involved in CG DNA methylation in poplar were stable over a 24 h period in leaves of P. nigra N46 compared with circadian-controlled genes. That could be part of the reason that methylation of CCGG sites is stable in those leaves. That DNA methylation differed even in genetically identical plants indicates the specificity of DNA methylation changes in their genomes. No statistically significant differences in methylation changes were found between day and night, suggesting that DNA methylation is more stable than expected and is unlikely to be involved in circadian regulation in plants.

Transformation of Tobacco with Two AP1 Genes Isolated from Poplar( Populus simonii × Populus nigra)

[ Objective ] This study aimed to analyze the functions of AP1 gene from Populus simonii × Populus nigra and to lay the theoretical foundation for shortening the breeding cycle of forest trees and investigating the flowering mechanism in poplar. [ Method] Plant expression vectors of AP1 genes were constructed and transformed into tobacco leaf disks with Agrobacterium-mediated method. Transgenic tobacco plants were identified by PCR. [ Result] AP1 genes were integrated into the genome of tobacco. Transgenic tobacco plants all presented an early flowering phenotype compared with wild-type tobacco. [ Conclusion] AP1 genes could promote early flowering in transgenic tobacco plants, which provided theoretical basis for molecular regulation of flowering in poplar.

The effect of number of joints on bending properties of laminated lumber made from poplar (Populus nigra)

Connection plays an important role in structural components and end jointing is one of the most common connections in structural applications. This study was designed to investigate some mechanical properties （MOE and MOR） of solid wood samples manufactured from poplar （Populus nigra）, joined together through end jointing. As well, we studied combinations of lumber manufactured from 10-mm veneer, using a polyvinyl acetate adhesive. The results show that non-jointed lumber （control samples） have better bending properties （MOE and MOR） than jointed specimens and lumber made from laminated layers. On the whole, side-by- side joints have a negative effect on the MOR and MOE of laminated products, which is more pronounced in the MOR. By increasing the number of joints, the properties of combination of three-ply, i.e., bending strength and modulus of elasticity, decreased.

Radial profile of sap flow velocity in mature Xinjiang poplar(Populus alba L. var. pyramidalis) in Northwest China

Estimation of the transpiration rate for a tree is generally based on sap flow measurements within the hydro-active stem xylem. In this study, radial variation of sap flow velocity（Js） was investigated at five depths of the xylem（1, 2, 3, 5 and 8 cm under the cambium） in three mature Xinjiang poplar（Populus alba L. var. pyramidalis） trees grown at the Gansu Minqin National Studies Station for Desert Steppe Ecosystem from May to October 2011. Thermal dissipation probes of various lengths manufactured according to the Granier＇s design were installed into each tree for simultaneous observation of the radial patterns of Js through the xylem. The radial patterns were found to fit the four-parameter GaussAmp equation. The peak Js was about 27.02±0.95 kg/（dm2？d） at approximately 3 to 5 cm deep from the cambium of the three trees,and the lowest Js appeared at 1 cm deep in most of the time. Approximately 50% of the total sap flow in Xinjiang poplar occurred within one-third of the xylem from its outer radius, whereas 90% of the total sap flow occurred within two-fifth of the xylem. In addition, the innermost point of the xylem（at 8-cm depth）, which appeared as the penultimate sap flow in most cases during the study period, was hydro-active with Js,8 of 7.55±3.83 kg/（dm2？d）. The radial pattern of Js was found to be steeper in midday than in other time of the day, and steeper diurnal fluctuations were recorded in June, July and August（the mid-growing season）. Maximum differences between the lowest Js（Js,1 or Js,8） and the highest Js（Js,3 or Js,5） from May through October were 12.41, 17.35, 16.30, 18.52, 12.60 and 16.04 g/（cm2？h）, respectively. The time-dependent changes of Js along the radial profile（except at 1-cm depth） were strongly related to the reference evapotranspiration（ET0）. Due to significant radial variability of Js, the mean daily sap flow at the whole-tree level could be over-estimated by up to 29.69% when only a single probe at depth of 2 cm was used. However, the accuracy of the estimation of sap flow in Xinjiang poplar could be significantly improved using a correction coefficient of 0.885.

黑桑(Morus nigra L.)的研究与利用进展

黑桑(Morus nigra L.)是桑属的一个种,具有悠久的栽培应用历史。黑桑不仅存在极为罕见的天然二十二倍体,而且其叶、茎、果中含有丰富的活性次生物质,具有抗氧化、抗炎、降血糖、美白等功能。本文系统总结关于黑桑的起源、分布、生物学特征、繁殖技术以及活性物质的功能和开发利用的研究进展,对上述研究中存在的一些争议进行了讨论,并提出今后对黑桑资源综合利用的主要研究方向和研究内容,以利于对黑桑种质资源的深入研究和更好地服务于我国黑桑的规模化栽培及资源的高效开发利用。

Differential protein expression in substantia nigra induced by 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine in a mouse model of chronic Parkinson’s disease

BACKGROUND： To date, a complete protein expression profile of the midbrain substantia nigra in a mouse model of chronic Parkinson＇s disease, induced by 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine （MPTP）, does not exist. In addition, there are no reports of analysis of differential protein expression. OBJECTIVE： To separate and evaluate MPTP-induced differential protein expression through the use of proteomics in the substantia nigra of a mouse model of chronic Parkinson＇s disease. DESIGN： Randomized controlled animal study. SETTING： Department of Neurology, the First Affiliated Hospital, Chongqing Medical University. MATERIALS： Sixteen 8-10-week old, healthy, male, C57BL mice, weighing 20-25 g, and of clean grade, were provided by the Experimental Animal Center of Chongqing Medical University. The experimental animals were disposed according to ethical criteria. MPTP was provided by Sigma Company, USA; Pdquest 2D image analysis software and gelatum/irradiance image analysis system （ChemiDoc XRS） by Bio-Rad, USA; and Voyager DE-PROMALD1-TOF-MS mass spectroscopy analyzer by AB1 Company, USA. METHODS： This study was performed in Chongqing Neurological Laboratory between November 2006 and July 2007. Mice were randomly divided into model and control groups, with 8 mice in each group. Mice in the model group were received a subcutaneous injection of MPTP （25 mg＆g）, twice a week, for five successive weeks, to establish a chronic Parkinson＇s disease model. Mice in the control group received the same volume of a subcutaneous saline injection at the same time points. Mice were sacrificed by anesthesia to rapidly obtain the midbrain for protein separation of the substantia nigra. MAIN OUTCOME MEASURES： （1） 2-ED handbook （Bio-Rad Company） was referenced for two-dimensional electrophoresis, （2） PDQUEST8,0 analytical electrophoresis pattern was adopted to evaluate differential protein expression. （3） Peptide mass finger print map and data were retrieved on http：//www.prospector.ucsf.edu to compare differential substantia nigral protein expression in the two groups. RESULTS： Two-dimensional gel electrophoresis of substantia nigra tissue indicated that there were 33 differential protein expressions between the two groups. Three new proteins were evaluated, including α -enolase, which exhibited regulated expression, tumor necrosis factor ligand superfamily member 4, and cyclin-dependent kinase inhibitor 1B. CONCLUSION： There are three proteins that exhibit differential expression in the substantia nigra- α -enolase, tumor necrosis factor ligand superfamily member 4, and cyclin-dependent kinase inhibitor 1B.

Seasonal Changes in Copper and Cobalt Concentrations of PinusnigraL.,Cedrus libani and Cupressus arizonica Leaves to Monitor the Effects of Pollution in Elazig (Turkey)

The aim of this study is to examine seasonal changes in Cu and Co concentrations of three plant species for monitoring the effects of pollution in Elazig,Turkey.For this purpose,the leaves of the Pinus nigra L.,Cedrus libani and Cupressus arizonica together with soil samples were collected from different points depending on traffic intensity,nearness the city center and cement factory as well as control location during different months of the year.Flame atomic absorption spectrophotometer(FAAS) was used for measurement of the metals in clear digests after the dry ashing method.Copper and Co concentrations were in the ranges from 1.3to 2.6mg.kg-1 and<LOD to 0.26mg.kg-1 for Pinus nigra L.,1.2to 4.7mg.kg-1 and<LOD to 0.41mg.kg-1 for Cedrus libani and 1.5to 4.8mg.kg-1 and<LOD to 0.42mg.kg-1 for Cupressus arizonica,respectively.The levels observed for Cu and Co in the soil ranged from 12to 38mg.kg-1 and 6.0to 17mg.kg-1,respectively.

转TaLEA基因小黑杨株系变异及生长稳定性分析

以6个地点栽培的11个转TaLEA基因小黑杨株系及1个对照株系为材料,对2年生株系的树高和地径进行调查分析。方差分析结果表明:树高、地径在不同地点间、不同株系间的差异达极显著水平(P<0.01),株系×地点间的交互作用达显著水平(P<0.05);不同地点12个小黑杨转基因株系2年生树高平均值变化范围为128～235cm,地径变化范围为13～24mm;不同地点参试株系树高和地径变异系数变化范围分别为19.84%～33.38%和24.21%～49.16%;重复力变化范围为0.497～0.952,表明相同地点不同株系树高和地径差异较大,但差异主要由遗传因素控制,有利于株系选择。采用Tai模型对各株系的遗传稳定性进行评价,其中XL11株系为不稳定株系,其他11个为稳定性较强的株系。根据各株系的稳定性参数和树高与环境的交互作用效应值,预测了各株系的适生地区,其中XL9、XL1、XL14等3个株系在6个试验地点的生长量大、稳定性强,是6个试验点推广的首选株系。

小黑杨FLC基因的克隆及功能解析

MADS-box家族蛋白是一类重要的调控开花的转录因子，FLC是其家族成员的编码基因之一。FLC在调节开花过程，其功能的发挥受多个途径的调控。本研究利用RT-PCR方法在小黑杨雄花芽中克隆出FLC基因的全长cDNA序列PnFLC。该基因的开放读码框为726bp，编码241个氨基酸残基组成的分子量为27．559kD的蛋白质，蛋白质的等电点为9．37。实时定量荧光PCR结果显示，春化能够使PnFLC在小黑杨根、茎、叶各组织中表达量下调57．9％～84％；启动子GUS染色结果表明，PnFLC启动子在分裂活跃的组织中活性较高。拟南芥的遗传转化结果显示，PnFLC可以调控AtAP1、AtSOC1和AtFT基因的表达，同时延迟拟南芥的开花时间。

欧美杂交杨Pnd-LRR3基因克隆及其抗锈菌侵染表达

为深入探究欧美杂交杨(Populus nigra×P.deltoides)感病的分子机理,以及抗病基因在发病过程中所起的作用,克隆了欧美杂交杨LRR3蛋白编码基因Pnd-LRR3。对Pnd-LRR3基因进行了生物信息学分析,并对其在抗锈菌过程中的功能进行了研究。Q-PCR结果显示,强致病性的落叶松-杨栅锈菌(Melampsora larici-populina)E4菌株接种6 h,Pnd-LRR3基因表达量上调增幅较大,168 h为显著下调。说明Pnd-LRR3基因在E4侵染过程中起到一定抗性作用,但最终无法阻止锈菌的侵染。

转TaLEA基因小黑杨抗寒株系的筛选

为验证转TaLEA基因小黑杨耐低温能力并筛选抗寒的优良株系,对11个转LEA基因小黑杨株系与非转基因野生型小黑杨对照(WT)进行了低温胁迫处理,测定其丙二醛(MDA)的质量摩尔浓度、过氧化物酶(POD)活性、过氧化氢酶(CAT)活性、叶绿素质量分数、冷害指数等。结果表明:低温胁迫条件下,除XL09外,其余各转基因株系的MDA质量摩尔浓度均低于WT,转基因株系MDA的质量摩尔浓度平均值低于WT的16.5%;抗氧化酶类活性在低温胁迫下仅XL10和XL11株系相对较高;叶绿素质量分数除XL01略低于WT外,其余转基因株系均高于WT,转基因株系叶绿素的质量分数平均值为35.1 mg.g-1,高于WT的6.4%;冷害指数方面,各转基因株系中除XL03高于WT外,其余均低于WT。采用隶属函数法综合评价各株系的抗寒性,从强到弱排序为:XL11>XL10>XL14>XL04>XL07>XL03>XL13>WT>XL06>XL01>XL05>XL09。外源基因LEA的导入不同程度地提高了小黑杨的抗寒性,其中XL11、XL10、XL14、XL04株系耐低温能力较强。

9种杨属植物雄花序中黄酮类含量的HPLC法测定

杨树(杨柳科Ｓａｌｉｃａｃｅａｅ杨属ＰｏｐｕｌｕｓＬ.植物)树皮、雄花序、芽脂、叶等均可药用。1977年版《中国药典》收载加杨(Ｐｏｐｕｌｕｓ×ｃａｎａｄｅｎｓｉｓＭｏｅｎｃｈ)、毛白杨(ＰｏｐｕｌｕｓｔｏｍｅｎｔｏｓａＣａｒｒ.)及杨属其他植物的雄花序,用于治?..

NaCl胁迫对欧洲黑杨组培植株叶片光系统Ⅱ功能的影响

为研究植物叶绿素荧光活性与耐盐性的关系 ,培育和选育抗盐植株 ,以组培无菌小植株为材料 ,研究了NaCl胁迫下黑杨叶绿素荧光诱导动力学的变化。测定结果表明 ,NaCl胁迫降低了欧洲黑杨试管内组培植株叶片PSⅡ (光系统Ⅱ)潜在活性 (FV/Fo)、原初光能转化效率 (Fv/Fm)和光合作用潜在量子转化效率 (Fd/Fs) ;降低了荧光光化学猝灭系数 (qP) ,提高了荧光非光化学猝灭系数 (qN) ,抑制叶绿体所捕获的光能用于光合作用 。

HPLC-ELSD法测定新疆药桑叶中1-脱氧野尻霉素的含量

该研究采用高效液相色谱-蒸发光散射检测(HPLC-ELSD)法测定新疆药桑叶中1-脱氧野尻霉素(1-DNJ)的含量,色谱柱为Waters XBridge^(TM) HILIC(2.1 mm×100 mm,5μm),乙腈-水(85∶15,V/V)为流动相,流速0.2 mL/min。结果表明,1-DNJ在1.2~12.0μg/mL质量浓度范围内线性关系良好,平均回收率为98.25%,相对标准偏差(RSD)为1.16%。该方法快捷简便、结果可靠,可用于药桑叶中1-脱氧野尻霉素含量的分析检测。

黑桑种质资源表型性状变异特征分析

【目的】研究新疆东南部21个采集点的710份黑桑种质资源表型性状,分析遗传多样性,为新疆黑桑种质资源的系统研究及开发利用奠定基础。【方法】对黑桑的15个定性描述性状和12个定量描述性状进行遗传多样性、主成分、聚类和相关性分析。【结果】叶片大小、枝条围度、叶幅、叶形等4个性状多样性较为丰富,其余定量描述性状多样性较低。黑桑表型性状聚类结果主要集中在两大类群,根据行政区域分布,一个类群主要集中在环塔里木盆地南缘县(市),另一个类群主要集中在环塔里木盆地北缘县(市)。果形指数与纬度之间呈显著负相关,其他定量描述性状与地理因子(经纬度和海拔)之间均无相关性,叶片大小和叶片周长的差异是区分不同地区黑桑品种的主要性状特征,此外,叶片大小和周长受到温度的正调控和降雨量的负调控。【结论】黑桑表型性状的遗传多样性较低。