Duplication of Locus Coding of Malate Dehydrogenase in Populus tomentosa Carr.

Horizontal starch-gel electrophoresis was used to study crude enzyme extraction from young leaves of 234 clones of Populus tomentosa Cart. selected from nine provenances in North China. Ten enzyme systems were resolved. One hundred and fifty-six clones showing unusual allozyme band patterns at locus Mdh-Ⅰ were found. Three allozyme bands at locus Mdh-Ⅰ were 9：6：1 in concentration. Further studies on the electrophoretic patterns of ground mixed pollen extraction of 30 male clones selected at random from the 156 clones were conducted and it was found that allozyme bands at locus Mdh-Ⅰ were composed of two dark-stained bands and a weak band. Only one group of the malate dehydrogenase （MDH） zymogram composed of two bands was obtained from the electrophoretic segregation of pollen leachate of the same clones. A comparison of the electrophoretic patterns one another suggested that the locus Mdh-Ⅰ coding malate dehydrogenase in diploid species of P. tomentosa was duplicated. The duplicate gene locus possessed three same alleles and was located in mitochondria. The locus duplication of alleles coding malate dehydrogenase in P.tomentosa was discovered and reported for the first time.     

Variation of leaf characteristics in Populus tomentosa Carr.

An investigation was conducted to determine the extent of variations among nine provenances of Populus tomentosa Carr. in terms of leaf characteristics. A total of 263 accessions were studied under field conditions in the National Gene Bank of P. tomentosa in 2003. All of the accessions were characterized by 17 indices from 1 to 2-dimension constructions. Variance analysis of all characteristics showed that there were significant differences among the nine provenances and among individuals within each provenance. This study reveals that the evaluated germplasm appears to have a wide genetic base and high potential for further genetic improvements and it also indicates that abundant gene resources of P. tomentosa have been collected and preserved in the National Gene Bank. [Supported by the “Tenth Five-year Plan” National Key Project in Science and Technology (Grant No. 2002BA515B0303) and the National “863” Project (Grant No. 2002AA241071)]     

Lignin characterization of triploid clones of Populus tomentosa Carr.

In order to understand the structural characteristics of lignin in triploid clones of Populus tomentosa and its changes in the processes of pulping and bleaching, milled wood lignin (MWL), lignin carbohydrate complex (LCC) and the residual lignin from kraft pulp (KP) and sulfite pulp (SP) were isolated and analyzed by Fourier transform infrared (FTIR) spectrum and ¹³C nuclear magnetic resonance (NMR). The most diagnostic peaks were assigned and the differences were discussed. The spectral patterns reveal that triploid P. tomentosa shows the specific features of hardwood from temperate areas, but in the spectrum of FTIR, the strength ratio of A 1270 cm⁻¹ to A1226 cm⁻¹ is 0.88, higher than the average of hardwood from temperate areas, which will make the lignin delignification more difficult during pulping and bleaching. The LCC from triploid P. tomentosa is mainly composed of xyloglucan and glucuronic acid, and other glucides have much lower ratio. In LCC FTIR, there are three peaks at 1 427, 1 329 and 1 046 cm⁻¹, indicating that both semi-cellulose and cellulose could exist in LCC, and that there might be relationships between cellulose and lignin. Compared with the residual lignin from KP and SP, the condensed structure in KP is more than that in SP.     

毛白杨优树无性系相关选择研究*

从全国选出的毛白杨优树中选择树龄５０年生以上的老树５５株，采用根繁获得的３年生无性系作为研究材料。通过方差分析估算了个体的重复力并分析了性状的稳定性；通过相关分析，特别是相关遗传力的通径分析，找出了影响苗木材积生长的主要性状，进而获得了优良基因型的表型标志。叶干重大、叶片窄而长、叶缘锯齿大而疏、叶柄较长、枝角适中（４５°左右）、枝数较多是毛白杨速生的主要特征。枝角和叶基用较大是毛白杨木纤维长度较长的标志之一。     

毛白杨优良外生菌根菌的选择*

在毛白杨外生菌根菌生态学特性和共生关系研究的基础上，对与毛白杨有良好共生关系的外个菌根菌的研究表明，硬皮马勃能耐温，在碱笥条件下仍能生长，在中性基质中，均能与毛白杨形成菌根，并大幅度地提高毛白杨的生长量。但在碱性土壤中，仅有硬皮马勃能与毛白杨形成一定数量的菌根，并能明显地提高毛白杨的生长量、地上部分生物量，改善菌木Ｎ、Ｐ的营养状况，接种效果随接种量的变化而差异明显。     

烟草和毛白杨次生维管发育相关MYB转录因子分析

[目的]为探讨额河杨和银灰杨天然杂种的起源机制,[方法]应用18对SSR标记,从分子水平上对新疆额尔齐斯河流域杨属植物的种间关系进行分析研究。[结果]表明:(1)SSR系统发育树将整个流域天然杨属植物分为两大类群,即黑杨派和青杨派为一类,白杨派为一类;(2)白杨派派内系统聚类图显示,银白杨、欧洲山杨、银灰杨三个树种均有较大的遗传分化,特别是杂种银灰杨似乎更大;(3)黑杨派和青杨派的UPGMA分类图显示,青杨派和黑杨派分属于2个分支,其中,青杨派内部分化相对简单,分为2支,均为典型的苦杨;黑杨派内部的分化较为复杂,可分为4类,包括典型的欧洲黑杨、额河杨和回交子代。[结论]杂种额河杨具有更多的欧洲黑杨的遗传成分,因此,将额河杨放到黑杨派是正确的。     

Cloning and RNAi construction of a LEAFY homologous gene from Populus tomentosa and preliminary study in tobacco

PtLFY, a LEAFY (LFY) gene, was cloned from Populus tomentosa (LM50) by PCR. Sequencing analysis indicated that PtLFY was 2 629 bp long, composed of three exons and two introns and encoded 378 amino acids. The splice donor sites and the splice acceptor sites were in identical positions to the LFY and its homologues. The amino acid sequence inferred was 68%-99% homologous to those of LFY and its homologues by blast analysis in GenBank. The Southern blot analysis indicated that there was a single copy of the PtLFY gene in genomic DNA of male and female P. tomentosa (LM50 and 5082). The pBI121-Ptalfy (reverse)-intron-Ptlfy-GUS-nos was constructed using RNA interference (RNAi) technique and verified by PCR and digestion identification and transformed into tobacco. Some transgenic tobacco plants were obtained by PCR and PCR-Southern identification. The growth was generally repressed in transgenic tobacco plants compared with wild-type ones and some phenotypic differences were observed. [Supported by the National Natural Science Foundation of China (Grant No. 30371175) and Postdoctoral Foundation of China (Grant No. 2002032041)]     

长期施氮与灌溉对毛白杨人工林土壤硝态氮分布与积累的影响

目的 研究灌溉和施氮对硝态氮在毛白杨林地土壤中积累与分布的影响,为毛白杨速生丰产林科学精准施肥和减少硝态氮淋失提供依据。 方法 在华北平原毛白杨适生地河北威县,利用"十二五"毛白杨大径材培育研究长期试验地,研究了通过灌溉保持不同土壤水分状况（田间持水量45%、60%、75%）和施氮量（0.0、101.6、203.2、304.8 kg·hm-2）对土壤硝态氮分布与积累的影响。 结果 土壤硝态氮分布具有明显的表聚性,在0200 cm土层呈先下降后上升再下降的"S"型变化趋势,且施氮量越大,硝态氮在60100 cm土层累积的趋势越明显。随着土壤水分的增加,土壤硝态氮的淋洗峰所在土层深度分别在4060、6080、80100 cm。土壤硝态氮累积量随施氮量的增加而增加,随灌水量的增加则呈先增加后减少的趋势;水氮交互作用对硝态氮的分布与积累影响显著。林地不同水氮组合土壤硝态氮积累量为86.11259.29 kg·hm-2。 结论 施氮量与土壤水分含量对毛白杨林地土壤硝态氮的分布与积累影响显著,在0200 cm土层内,随着施氮量的增加,硝态氮的积累量呈上升趋势;随着土壤含水量的增加,硝态氮积累峰所在的土层呈逐渐下移的趋势。与农田生态系统相比,试验林地0200 cm土层内硝态氮积累量较低,林地生态系统庞大林木根系对硝态氮吸收作用值得重视。同时,为了减少硝态氮的淋失,建议减少氮肥的施用,并将田间持水量控制在60% 75%。     

Molecular cloning of a cellulose synthase gene PtoCesA1 from Populus tomentosa and its genetic transformation in tobacco

A 3 125 bp cellulose synthase gene, PtoCesA1, which has a 98% identity to PtrCesA1 from Populus tremuloides, was cloned from cDNA prepared from secondary xylem of P. tomentosa. Four anti-expression vectors with different fragments of PtoCesA1, named as pBIPF, pBICC1, pBIPR and pBIBR, were constructed. Some traits of transformed tobacco of pBICC1, pBIPR and pBIBR differed from wild types, such as small leaves, “dwarf” phenotype and thinner xylem and fiber cell walls than wild plants consistent with a loss of cellulose. It indicated that the growth of transgenic tobacco was restrained by the expression of anti-PtoCesA1. Transgenic tobacco was obtained and the contents of cellulose and lignin were analyzed as well as the width and length of fiber cells, and xylem thickness for both transgenic and control plants. Transformed tobacco showed a different phenotype from control plants and it implied that PtoCesA1 was essential for the cellulose biosynthesis in poplar stems. [Supported by the Hi-Tech Research and Development Program of China (863) (2001AA244060 and 2003AA244020) and National Basic Research Program of China (973) (J1999016003)]     

Variation of Leaf Characteristics in Populus tomentosa Carr.

An investigation was conducted to determine the extent of variations among nine provenances of Populus tomentosa Carr. in terms of leaf characteristics. A total of 263 accessions were studied under field conditions in the National Gene Bank of P. tomentosa in 2003. All of the accessions were characterized by 17 indices from 1 to 2-dimension constructions. Variance analysis of all characteristics showed that there were significant differences among the nine provenances and among individuals within each provenance. This study reveals that the evaluated germplasm appears to have a wide genetic base and high potential for further genetic improvements and it also indicates that abundant gene resources of P. tomentosa have been collected and preserved in the National Gene Bank.     

毛白杨无性系树体性状的关系分析

对１０年生毛白杨无性系５个树体性状组的关系分析结果表明，材积、枝下高、圆满度三个重要经济性状的改良潜力较大，通直度的改良效果较差。主干和侧枝两性状组与经济性状组的关系密切，分别解释了经济性状组５４７５％和２８７６％的相关变异。对经济性状实施选择时，可把树高、胸径、Ｈ／２径、冠幅、侧枝的数量与密度、主枝的长与粗以及新枝长和节数作为间接选择指标。     

毛白杨苗期VA菌根菌与外生菌根菌接种的研究

1998～1999年在田间条件下布置了毛白杨VA菌根(包括根内球囊霉菌和单孢球囊霉菌)与外生菌根硬皮马勃混合接种试验.结果表明接种菌根菌对毛白杨苗高生长量有促进作用,且以根内球囊霉菌作用最明显,单孢球囊霉菌处理次之,硬皮马勃处理最差.水涝大大降低人工接种菌根菌的效果,严重影响VA菌根和外生菌根菌单独接种及混合接种的效应,根内球囊霉菌甚至会抑制毛白杨苗木的高生长,VA菌根和外生菌根间的负交互作用也不对苗高生长产生影响.菌根真菌的感染率随时间变化出现较大的波动和变化.     

胡杨分子生物学研究进展

胡杨具有抗盐碱、抗旱等特性,是极端干旱沙漠地区唯一的乔木树种,对于维持荒漠地区生态平衡具有重要作用。胡杨对多种逆境的高抗性和良好适应性成为近年来研究的热点。关于胡杨分子生物学方面的研究,相对于其他模式植物研究开展较晚。文中综述了近年来国内外胡杨分子生物学研究的阶段性进展,包括分子标记开发、遗传多样性检测、基因组和转录组的研究以及重要的抗逆相关功能基因的分析等;讨论了目前胡杨分子水平研究存在的不足,并对胡杨分子生物学研究的发展前景作出展望,旨在为胡杨的保护和育种提供更多的科学参考。     

超量表达FBL1对84K杨根系和生长量影响研究

生长素及其信号转导系统对植物的生长发育具有重要的影响。本研究从银腺杨'84K'(Populus alba × P. glandulosa cl. '84K')中分离了生长素受体基因PtrFBL1,利用PMDC32构建了PMDC32-PtrFBL1超量表达载体,并通过遗传转化获得了超量表达植株17个。对温室定植的3个转基因株系和对照植株的根系、生长量和光合指标等性状分析结果显示:转基因株系总根长和总根面积达到显著或极显著差异,而根系干质量、平均不定根系长度、平均不定根直径差异不显著;株高、平均节间长、地径和高径比皆高于对照,且大多数转基因株系达到显著差异;除气孔限制值(Ls)低于对照外,气孔导度(Cd)、水分利用效率(WUE)、光能利用效率(LUE)和叶绿素相对含量皆高于对照,且大多数转基因株系达到显著或极显著差异。以上结果表明,可能是FBL1超表达增加了转基因株系根系面积,提高了水分和养分的吸收利用,进而导致转基因株系光能吸收和转化效率提高,引起转基因株系生长加快。     

实验田4年生转基因毛白杨嫁接苗和自根苗安全性研究

目的 对种植于天津滨海的自根苗转AhDREB1基因毛白杨（（Populus tomentosa × P. bolleana） × P. tomentosa）以及种植于沧州盐山的嫁接苗转基因株系进行安全性研究,评估转基因毛白杨对环境可能造成的影响。 方法 利用PCR扩增以及电泳技术,对种植在天津滨海和沧州盐山实验田的4年生转基因毛白杨基因组DNA、土壤DNA、沧州嫁接苗砧木DNA以及抗性微生物基因组DNA进行PCR特异性检测。对天津实验田土壤中根际微生物数量进行连续动态监测（3月、4月、5月）。分别在天津滨海和沧州盐山实验田模拟自然条件下转基因毛白杨植株枯落物掉落,检测不同生长阶段的不同器官（新生嫩芽、多年生枝条、新生根尖、多年生侧根、新生嫩叶、当年生老叶）的降解时间。通过化感实验,检测天津滨海和沧州盐山实验田转基因毛白杨植株叶片对白菜（Brassica pekinensis）种子的生长是否造成影响。 结果 电泳结果显示：外源基因稳定存在于转基因毛白杨基因组中,实验田土壤DNA、抗性微生物DNA样品以及沧州嫁接苗砧木DNA中均未出现目的基因片段;天津非转基因植株和转基因植株的根际土壤中可培养微生物数量没有显著差异;转基因毛白杨的枯落物,无论是飘落在杂草表面、土壤表面还是埋入土壤,2个月后外源基因均被降解。化感实验结果显示：非转基因植株以及转基因植株对白菜种子的胚轴和胚根生长的影响没有显著差异。 结论 未发现实验田种植4 a的转基因毛白杨对周围的环境造成影响。     

Characterization and role of glucose-6-phosphate dehydrogenase of Populus suaveolens in induction of freezing resistance

Glucose-6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49) was purified from the leaves of 8-week-old Populus suaveolens cuttings. The enzyme activity in the absence and presence of reduced dithiothreitol (DTT_red) was determined. The results show that the G6PDH activity is not inactivated by pre-incubation with DTT_red, indicating that the purified enzyme probably presented in cytosol of P. suaveolens. The catalytic characteristics and kinetic parameters of cytosolic G6PDH purified from P. suaveolens cuttings were also studied. The results show that G6PDH is characterized by K _m value of 360 μmol·L⁻¹ for G6P and 16 μmol·L⁻¹ for NADP, a pH range of 7.3-8.9, and the maximum activity around pH 8.2. The enzyme activity is inhibited by various metabolites such as NADPH, NADH, GTP, UTP, ATP, AMP, ADP, CoA, acetyl CoA, fructose-6-phosphate (F6P), erythrose-4-phosphate (E4P), ribose-5-phosphate (R5P) and 3-phosphoglycerate (3-PG) (all at 1 mmol·L⁻¹ except for NADPH and NADH) to different extents. NADPH is the most effective inhibitor of enzyme activity, with an inhibition of 72.0%. The addition of metal ions such as MgCl₂, CaCl₂ and KCl (all 1.0 mmol·L⁻¹) to the standard reaction mixture has no remarkable influence on the cytosolic G6PDH activity. However, CdCl₂ (1.0 mmol·L⁻¹) causes high inhibitory effect on the enzyme activity. To explore the role of G6PDH on the enhancement of freezing resistance induced by freezing acclimation, the changes in the cytosolic G6PDH activity and freezing resistance (expressed as LT₅₀) of P. suaveolens cuttings during freezing acclimation at −20 °C were investigated. The results reveal that freezing acclimation decreases LT₅₀ of cuttings, and increases the activity of cytosolic G6PDH compared with control ones, while 2 d of de-acclimation at 25 °C result in a decrease in cytosolic G6PDH activity, and caused an increase in LT₅₀. Furthermore, the change in cytosolic G6PDH activity is found to be closely correlated to the degree of freezing resistance of cuttings during freezing acclimation. It is suggested that cytosolic G6PDH may be involved in the induction of freezing resistance of cuttings. [Supported by the Foundation of State-designated Base for Biology Researching and Teaching in Beijing Forestry University]     

Interrelationships between leaf heat conductivity and tissue structures of different varieties of Populus tomentosa Carr.

Plant heat conductivity largely depends on tissue structure. Different structures lead to different heat conductivity. As well, water transfer also plays a very important role in heat transfer in plants. We have studied leaf heat conductivity and tissue structure of 3- and 30-year-old Populus tomentosa Carr. trees using mildred thermal imaging, steady state heat conductivity surveys and paraffin section and investigated the relationship between leaf heat conductivity, tissue structure and water content of leaves. The results show that the temperature on leaf surfaces among the various varieties of trees was almost the same. Leaf heat conductivity, temperature and water content of leaves are positively correlated. The thicker the leaf tissue structures, the larger the heat resistance. That is, the tighter the cells and the smaller the interspaces, the smaller the heat conductivity, which is not conducive for heat transfer.     

胡杨基因组片段转化拟南芥表型研究

[目的]本研究旨在探索与挖掘胡杨基因组大片段的潜在功能,发掘具有潜在育种价值的胡杨基因簇。[方法]利用已构建的胡杨基因组BIBAC文库,采用花序浸染法,将胡杨基因组大片段78A2D10导入模式植物拟南芥基因组中。采用抗性筛选、分子检测及表型观察等方法鉴定、分析转化型植株。[结果]共获得15株特异表型的转化植株。与野生型相比,转化型植株主侧茎生长受到抑制,莲座叶面积增大近3倍,叶片数量增多,叶边缘皱缩,抽薹推迟约13周,株高增加近32.0 cm,侧茎发育成次生莲座,植株寿命延长约7周。[结论]胡杨基因组片段78A2D10可延长植株营养生长期及植株寿命,据此推测该基因片段可能与营养生长有关。     

毛白杨无性系木材基本密度遗传变异研究(英文)

对河北、河南、山东、甘肃的４块毛白杨无性系测定林中约１００个无性系进行了木材基本密度的测定，对木材基本密度在地点和无性系间的遗传变异进行了研究．结果表明：毛白杨无性系的木材基本密度平均值为０．４３６３ｇ／ｃｍ３，木材基本密度在地点和无性系间均存在极显著差异．木材基本密度的无性系重复力为０．８２～０．９１，属于受遗传控制较强的材性性状．对生长和材性的相关研究表明，木材基本密度与生长性状（包括胸径、树高和材积）一般存在显著的负相关，但不同地点上表现为不同的正或负的相关关系     

Assessment of rhizospheric microorganisms of transgenic Populus tomentosa with cowpea trypsin inhibitor ( CpTI ) gene

To have a preliminary insight into biosafety of genetically transformed hybrid triploid poplars (Populus tomentosa × P. bolleana) × P. tomentosa with the cowpea trypsin inhibitor (CpTI) gene, two layers of rhizospheric soil (from 0 to 20 cm deep and from 20 to 40 cm deep, respectively) were collected for microorganism culture, counting assay and PCR analysis to assess the potential impact of transgenic poplars on non-target microorganism population and transgene dispersal. When the same soil layer of suspension stock solution was diluted at both 1:1 000 and 1:10 000 rates, there were no significant differences in bacterium colony numbers between the inoculation plates of both transgenic and non-transgenic poplars. The uniform results were revealed for both soil layer suspension solutions of identical poplars at both dilution rates except for non-transgenic poplars at 1:10000 dilution rates from the same type of soil. No significant variation in morphology of both Gram-positive and Gram-negative bacteria was observed under the microscope. The potential transgene dispersal from root exudates or fallen leaves to non-target microbes was repudiated by PCR analysis, in which no CpTI gene specific DNA band was amplified for 15 sites of transgenic rhizospheric soil samples. It can be concluded that transgenic poplar with the CpTI gene has no severe impact on rhizospheric microorganisms and is tentatively safe to surrounding soil micro-ecosystem. [Supported by the National Project in Transgenic Plant and Application (Grant No. J2002-2003)]