欧洲黑杨基因资源稳定碳同位素组成特征

通过测定从欧洲不同地区引进的134个欧洲黑杨无性系叶片的稳定碳同位素比率(δ13C值),研究了欧洲黑杨基因资源δ13C值的特点和水分利用状况.结果表明:(1)引进的欧洲黑杨基因资源叶片δ13C值为-30.40‰～-27.02‰,水分利用效率具有丰富的多样性,优于国内乡土树种,其中来自俄罗斯、土耳其和德国的基因资源δ13C值明显高于其它国家的基因资源,可为抗旱杨树新品种的选育提供优良的亲本材料;(2) 相同基因资源在不同地区栽植时,δ13C值受环境的影响较大.生长在内蒙古地区的欧洲黑杨无性系叶片平均δ13C值为-26.53‰,明显高于在陕西和北京地区测得的-28.39‰和-28.93‰,故在进行不同基因型的δ13C值评价和选择时要在相同条件下进行;(3)生长在不同地区的欧洲黑杨的δ13C值与生长量的相关程度不同,生长在北京地区的欧洲黑杨δ13C值与株高、地径呈极显著正相关(P＝0.01),相关系数分别为0.912和0.829,而生长在陕西和内蒙古地区的欧洲黑杨的δ13C值与生长指标虽然也相关,但达不到显著水平.在研究植物生长与水分利用效率之间的关系时,必须结合植物的生长环境进行分析.     

新疆额尔齐斯河流域苦杨与欧洲黑杨遗传多样性分析

应用SSR标记技术,对新疆额尔齐斯河流域河谷分布的苦杨(Populus laurifoli)和欧洲黑杨(Populus nigra)天然居群的遗传多样性和遗传分化进行了研究。结果显示:苦杨和欧洲黑杨的天然居群具有较高的遗传多样性,且苦杨明显高于欧洲黑杨。与苦杨居群相比,欧洲黑杨居群间基因流较高,居群间的遗传分化程度较小,变异主要源于居群内。苦杨和欧洲黑杨居群内的杂合度较高,各个居群的各项遗传参数都要小于物种水平。苦杨和欧洲黑杨均偏离Hardy-Weinberg平衡,表现为杂合子过量的现象。     

TA29-Barnase基因导致抗虫转基因欧洲黑杨雄性不育的研究

取在１／２ＭＳ＋ＮＡＡ０．０２ｍｇ／Ｌ培养基中发育４￣６周的欧洲黑杨转Ｂｔ基因植株１５－ｎ－１９２试管苗试叶,在ＭＳ＋ＢＡ０．２ｍｇ／Ｌ＋ＮＡＡ０．０２ｍｇ／Ｌ培养基中培养１６ｈ,分别用基因枪法和叶盘法转化ＴＡ２９－Ｂａｒｎａｓｅ嵌合基因,其Ｂａｒｎａｓｅ基因的转化率依次为１６．１％和２３％,再生株数分别为１７５／１２和７１／１０。试验表明,选择发育适宜的叶片,利于目的基因转化后的植株再生。进一步     

欧洲黑杨优质核心种质库的初步构建

对159份欧洲黑杨种质资源的生长、育种值、材性、养分利用率和水分利用率等13个性状进行分析,采用13种表型选优的抽样方法构建欧洲黑杨优质核心种质库。表型值评价表明：当抽样群体比例达到21.4%以上即能满足构建核心种质要求,而只有抽样群体比例达到30.2%以上 SSR 分子遗传多样性才能满足构建核心种质要求。最终确定的欧洲黑杨优质核心种质库包含48份种质资源,占原群体的30.2%;与原群体相比,均值和方差不存在显著差异,极差符合率和变异系数变化率分别为97.07%和89.72%;优质核心种质库的有效等位基因数、Nei’s 遗传多样性系数和 Shannon’s 信息指数与原群体之间没有显著差异,保持了原群体的遗传多样性。     

欧洲黑杨基因资源光合生理特征与生长的关系

测定来自欧洲不同地区和我国的108个欧洲黑杨无性系(基因型)气体交换、叶绿素荧光与生长特征参数,研究欧洲黑杨基因资源光合生理与生长性状的遗传变异及相关性.结果表明:欧洲黑杨基因资源内气体交换、叶绿素荧光特征和生长性状具有丰富的遗传变异;来自我国和英国的无性系具有较高的净光合速率(Pn),来自匈牙利的无性系PSII最大光化学效率(Fv/Fm)最高.各检测参数在欧洲黑杨中具有较高的广义遗传力且变异系数较高,气孔导度(Gs)和PSII电子传递速率(ETR)可能是影响欧洲黑杨光合能力的重要限制因素.相关分析结果发现,Pn、胞间二氧化碳浓度(Ci)、Gs和ETR等参数指标在欧洲黑杨光合效率评价中具有潜在的应用价值;起源于塞尔维亚及其他南欧、东欧的欧洲黑杨可在进一步的杨树高光效育种研究中作为优良基因型选择的重点.     

SSR标记与欧洲黑杨材性性状的关联分析

对引进的92个1年生和65个4年生欧洲黑杨无性系进行材性测定,并结合171个SSR标记进行材性数量性状位点的关联分析.结果表明:1年生和4年生欧洲黑杨的木材基本密度分别为0.308 8 g·cm-3和0.355 2 g·cm-3,纤维长分别为601.3 μm和649.4 μm,纤维宽分别为21.81 μm和23.11 μm,微纤丝角分别为26.5°和26.1°.4年生欧洲黑杨苯醇抽提物、综纤维素、α纤维素以及木质素含量分别为2.32%、78.62%、41.48%和20.93%.通过SSR标记位点与性状间的关联分析,共获得与1年生材性性状关联的SSR标记位点10个,其中基本密度3个,微纤丝角5个,纤维长2个,纤维宽4个,贡献率6.43%～23.18%;与4年生欧洲黑杨材性性状关联的SSR标记位点8个,其中基本密度、纤维长、木质素各1个,微纤丝角、综纤维素含量和α纤维素含量各2个,贡献率7.41%～28.56%.     

抗虫转基因欧洲黑杨的Western印迹法分析

抗虫转基因欧洲黑杨的Ｗｅｓｔｅｒｎ印迹法分析陈颖，李强，李玲，韩一凡，田颖川（中国林业科学研究院林业研究所北京１０００９１）（中国科学院微生物研究所北京１０００８０）关键词Ｂｔ．基因，转基因欧洲黑杨，Ｗｅｓｔｅｒｎ印迹法ＷＥＳＴＥＲＮＢＬＯＴＡＮＡＬ...     

转Bt基因欧洲黑杨抗虫稳定性及其对天敌昆虫的影响

1997-2001年,对新疆玛纳斯平原林场的转Bt基因欧洲黑杨试验林进行了食叶害虫(杨尺蠖)和杨梦尼夜蛾危害调查,转基因试验林叶片平均损失率均低于10%,试验林地虫口密度由1997年的18头·m-2降低到2001年的8头·m-2,而对照林地虫口密度却呈上升趋势. 2005年,位于北京怀柔的转Bt基因欧洲黑杨试验林受杨尺蠖危害,叶片损失率不超过20%,而当地其它杨树林叶片损失率高达90%.在转Bt基因欧洲黑杨林(新疆玛纳斯)中,寄生靶标害虫的天敌种类、数量和寄生率均高于对照林,对从试验林中采集的靶标昆虫的蛹进行接种试验,与对照相比,寄生蜂出蜂率和数量均无明显差异,寄生蜂也无异常,说明转Bt基因欧洲黑杨未对天敌昆虫产生明显不利影响.     

欧洲黑杨转Bt毒蛋白基因植株大田抗虫性测定

近几年,基因工程发展很快,通过基因工程已获得了各种各样的转基因植物,如抗虫转基因农作物的棉花（ＧｏｓｙｐｉｕｍｈｉｒｓｕｔｕｍＬ．）、水稻（ＯｒｙｚａｓａｔｉｖａＬ．）、马铃薯（ＳｏｌａｎｕｍｔｕｂｅｒｏｓｕｍＬ．）、玉米（ＺｅａｍａｙｓＬ．）、小麦...     

中国和外来杨属种质资源

介绍了中国天然分布和从国外引进的杨属(PopulusL.)种质资源。总结了中国天然分布的杨属种质资源的派别、种类、分布范围和分布特点。在中国杨属5派中,以青杨派(Tacamahaca)种类最多,白杨派(Populus,以前称Leuce)分布最为广泛,黑杨派(Aigeiros)最具经济价值。将中国杨属树种引种历史划分为解放前的零星引种时期、50年代至80年代的以引进国外培育成功的优良品种为主时期和90年代后注重种质资源引进时期三个阶段,介绍了每个阶段的引种特点和引进的种质资源。在过去50年种,中国天然分布的白杨派和引进的黑杨派(Aigeiros)种质资源在中国的杨树栽培和育种中发挥了巨大作用。     

杨树基因组SSR标记在康定柳中的通用性分析

SSR分子标记因具有共显性遗传、高度多态性、重复性和稳定性好、易于实现自动化等优点,在遗传育种等方面有着广泛的应用前景。本研究在杨树基因组中随机选取622对SSR引物对来自5个不同群体的康定柳雌雄各20个个体,共计200个进行了通用性分析。结果表明：杨树基因组SSR标记在康定柳中的通用性达到7.88%。     

油松毛虫亚居群遗传结构的SSR分析

采用SSR分子标记技术对平泉县5个油松毛虫亚居群进行遗传多样性和遗传分化研究.结果显示:8对SSR引物对5个油松毛虫亚居群的扩增片段长度范围为78～430 bp,检测到的等位基因数为1～6个;种群总体水平多态位点比率P=87.50%,平均有效基因数A＝3.1250,平均期望杂合度He＝0.474 7,种群平均遗传距离为0.070 3～0.419 7;遗传分化度Fst=0.215 9,基因流Nm=0.908 1.由此可以得出在5个天然油松毛虫亚居群中,天然油松纯林居群内杂合度要比其他居群高;油松毛虫亚居群间已发生明显的遗传分化,基因交流较少,遗传漂变已经成为导致该物种种群分化的主要原因之一;油松毛虫亚居群遗传结构产生变异推测是由于常年化学防治和单一寄主植物造成的选择压力的影响.     

Variation in juvenile traits of natural black poplar (Populus nigra L.) clones in Turkey

Natural black poplar (Populus nigra L.) clones sampled from river courses were tested in the arid southeast region of Turkey, using a randomized complete block field design with four replications. Clones were laid out in row plots of eight ramets each. Height and apical dominance were assessed at age one year; diameter, survival, bole straightness and branchiness were measured at age two years. Clones differed significantly in survival, growth and quality traits. The results showed that promising clones exist in natural populations. Two commercial clones out of four did not grow as fast as the top new selection clones. For bole straightness, three commercial clones had significantly lower grading scores than the top best 14 clones. Principal components analysis indicated that growth, apical dominance and branching are the most important traits distinguishing black poplar clones. Diameter had a moderate correlation (0.34) with bole straightness. Relationships between geographic variables (elevation, latitude and longitude) with growth and stem quality traits were weak. Considerable genetic variation was observed among clones for all the traits. Genetic differences among the clones accounted for 27% (survival, bole straightness) to 39% (height) of the total variance. Broad-sense individual heritability ranged from 0.27 (survival) to 0.37 (apical dominance). Clonal mean heritabilities were higher than individual heritabilities and ranged from 0.60 (survival) to 0.82 (diameter), implying considerable gain could be realized via selective improvement methods.     

利用荧光SSR标记鉴别杨树品种

世界上天然杨树约100多种,我国有50多种。杨树作为重要造林树种,多用于水土保持、防风防沙和园林绿化,也是造纸、建筑及木材工业的重要经济树种。杨树易繁殖,生长速度快,生产周期短,在我国广泛栽培,我国也是世界上杨树人工林面积最大的国家(Carleetal.,2005)。我国杨树引种和育种的历史可追溯到新中国建立时期,杨树种质资源十分丰富,我国已培育了许多优良杨树品种(马常耕,1995)。随着育种技术的发展,新品种产生的速度与日俱增。然而,由于杨树品种繁多,形态相似,容易出现品种混淆。特别是近年来杨树种苗市场的迅速发展,为了追逐经济效益,杨树种苗市场上出现了以次充好,甚至出现造假的现     

欧洲黑杨PnEXPA1基因多态性与稳定碳同位素比率的关联分析

对欧洲黑杨α-expansin基因PnEXPA1的SNP多态性与水分利用效率相关性状稳定碳同位素比率(δ13C)进行了关联分析。利用SNaPshot技术对PnEXPA1基因内11个SNP位点进行了基因型分型,发现各SNP位点优势基因型均为纯合,且其频率高于杂合基因型。关联分析显示:SNP8和SNP12在采用单因素方差分析(ANOVA)和一般线性模型(GLM)2种方法时均与δ13C值显著关联。SNP8为exon 1内的无义突变,可解释6.620%的表型变异;而SNP12位于intron 1中,遗传贡献率为6.613%;这2个SNP位点与SNP9、SNP13共同位于一个高连锁不平衡(LD)的单倍型块中。SNP8与SNP12的TT基因型无性系均具有较高的δ13C值,为优势基因型。     

Development and transferability of two multiplexes nSSR in Scots pine (Pinus sylvestris L.)

Single sequence repeat(SSR) multiplexing is a semi high-throughput PCR methodology for the analysis of multiple SSRs.We developed two SSR multiplexes selected from SSR loci previously reported in the pine literature and tested the transferability of both SSR multiplexes in nine other pine species.We tested 234 nuclear SSR loci(n SSRs) previously described in the pine literature and selected ten n SSRs following the simple criteria of interpretability and reproducibility.Selected nuclear loci were divided into two n SSRs multiplex sets and their amplification was optimized for three different multiplex PCR methods based on:(a) a custom PCR protocol,(b) a custom protocol with hotstart taq polymerase,and(c) a commercially available kit for SSR multiplexing.To validate their performance,the level of genetic diversity was assessed in three Scots pine natural populations(Hungary,northern Sweden and southern Sweden).In addition,we also tested the transferability of these multiplexes in nine other pine species.We have developed two n SSRs multiplexes of five loci each that will contribute to reduce the costs of n SSR scoring,while increasing the capacity of n SSR loci analysis.Amplification was successful in all three populations(94 % success) and the level of polymorphism(7.1 alleles/marker) was similar to that previously reported for other Scots pine natural populations.Transferability of both multiplexes was successful for those pine species closely related to Scots pine.     

ELISA Quantitation and GC-MS identification of abscisic acid in stigma, ovary and pedicel of pollinated poplar flowers (Populus nigra L.)

A method of fractionation, quantitation and identification of abscisic acid (ABA) in poplar flowers is presented in the context of intra- and interspecific pollination studies in Populus nigra x P. nigra and P. nigra x P. deltoides, respectively. Poplar flowers were dissected into stigma, ovary and pedicel. Abscisic acid concentrations were measured at receptivity and 5 days after pollination in each part of the flower. Abscisic acid was fractionated by reverse-phase HPLC followed by an enzyme-linked immunosorbent assay (ELISA) with anti-ABA polyclonal antibody and GC-MS identification of the immunoreactive ABA fractions. Samples were fractionated and ABA was identified by the presence of a cross-reactive metabolite of ABA in the extracts. The interspecific cross (P. nigra x P. deltoides) had a high concentration of ABA in the pedicel, where abscission occurred, whereas in the intraspecific cross, ABA concentration in the pedicel was low. The results are discussed with reference to the control ABA exerts on the abscission of poplar flowers after interspecific incompatible pollinations.