Identification of differentially expressed genes associated with bud dormancy release in tree peony （Paeonia suffruticosa） by suppression subtractive hybridization

A subtractive cDNA library was developed to study genes associated with bud dormancy release in tree peonies.In order to identify genes that are highly expressed in buds released from dormancy,588 clones were examined by differential screening.Of these,185 clones were selected to be sequenced.A total of 37 unique sequences were obtained of which only 31 sequences have matches in the NCBI database or the Arabidopsis thaliana protein database.Semi-quantitative RT-PCR was used to confirm further the expression...     

枣抑制消减cDNA文库构建与分析

枣树具有独特的开花结果特性,为获得枣花发育基因,以金丝4号枣不同发育时期的花蕾和同时期的叶片cDNA互为试验方和驱动方,利用抑制消减杂交技术分别构建了枣花和枣叶抑制消减杂交cDNA文库(SSH文库)。枣花和枣叶SSH文库重组率分别是92.05%和90.26%,插入片段长度均介于200~2 000 bp之间,主要集中于1 000 bp左右。分别从枣花和枣叶SSH文库中随机挑选1 000个阳性克隆进行DNA测序、组装拼接,结果显示枣花SSH文库中获得96条unigene,其中contigs 43个,singletons 53个,96条unigene预测得到73个ORF,经过同源比对分析获得有注释的序列20条,按功能分为7类;枣叶SSH文库中获得86条unigene,其中contigs 45个,singletons 41个,86条unigene预测得到70个ORF,经过同源比对分析获得有23条注释的序列,按功能分为6类。     

日本落叶松扦插生根过程中SSH文库构建及主要表达基因分析

为研究日本落叶松在扦插生根过程中基因的表达情况,以日本落叶松扦插生根率存在显著差异的2个优良全同胞无性系为材料,构建差异表达cDNA文库,获得正、反2个消减文库。随机挑取正库和反库各500个克隆进行测序,将测序结果整理后,正库得到272个UniEST,反库得到249个UniEST。这些UniEST分别属于新陈代谢、信号途径、物质运输、抗性相关、生长发育等类别基因,可能在扦插生根过程中具有重要作用。其中,获得10个与激素相关的基因,表明激素在日本落叶松扦插生根过程中发挥重要作用。除激素类基因外,还获得小G蛋白、ABC转运蛋白类、因伤诱导物质等相关基因。上述结果表明,日本落叶松扦插形成不定根是一个非常复杂的过程,在这个过程中激素相关基因起到关键作用,同时其它类别基因也发挥重要作用。     

Hydraulic characteristics and water relations in pigment-less mutant shoots of an orange tree

Phenotypes more or less deficient in photosynthetic pigments show reduced productivity. Not much is known, however, about the influence of pigment-less twigs on the water balance of whole trees. We studied the water relations and hydraulic properties of normal and pigment-less (white) and 1-year-old shoots of 12-year-old Citrus sinensis L. trees. Compared with green leaves, white leaves showed a pronounced deficiency of pigments, higher stomatal density, the absence of chloroplasts in the guard cells and a different organization of leaf parenchyma. Stomatal conductance (gL) and transpiration rate (EL) were higher in white leaves than in green leaves during the hottest hours of the day, especially in July and September. The absence of chloroplasts in the stomatal guard cells seemed to be one of the factors causing insufficient stomatal control. Hydraulic conductance (KL) was higher in white leaves than in green leaves (16.96+/-2.24x10(-5) versus 11.26+/-0.66x10(-5) kg s-1 m-2 MPa). The ratio between the sum of the fourth power of xylem conduit radius (Sigmar4) (which determines theoretical conductance) and the total leaf area supplied by petioles and midribs was higher in white leaves than in green leaves. This was because of a smaller leaf area in white leaves and a statistically different distribution of lumen diameters of the conduits in midribs and petioles. The hydraulic properties of white twigs profoundly disturbed the water balance and physiology of the whole tree.     

Operational flexibility in forest fire prevention and suppression: a spatially explicit intra-annual optimization analysis,considering prevention, (pre)suppression,and escape costs

Increasing wildfire threats and costs escalate the complexity of forest fire management challenges, which is grounded in complex interactions between ecological, social, economic, and policy factors. It is immersed in this difficult context that decision-makers must settle on an investment mix within a portfolio of available options, subject to limited funds and under great uncertainty. We model intra-annual fire management as a problem of multistage capacity investment in a portfolio of management resources, enabling fuel treatments and fire preparedness. We consider wildfires as the demand, with uncertainty in the severity of the fire season and in the occurrence, time, place, and severity of specific fires. We focus our analysis on the influence of changes in the volatility of wildfires and in the costs of escaped wildfires, on the postponement of capacity investment along the year, on the optimal budget, and on the investment mix. Using a hypothetical test landscape, we verify that the value of postponement increases significantly for scenarios of increased uncertainty (higher volatility) and higher escape costs, as also does the optimal budget (although not proportionally to the changes in the escape costs). Additionally, the suppression/prevention budget ratio is highly sensitive to changes in escape costs, while it remains mostly insensitive to changes in volatility. Furthermore, we show the policy implications of these findings at operational (e.g., spatial solutions) and strategic levels (e.g., climate change). Exploring the impact of increasing escape costs in the optimal investment mix, we identified in our instances four qualitative system stages, which can be related to specific socioecological contexts and used as the basis for policy (re)design. In addition to questioning some popular myths, our results highlight the value of fuel treatments and the contextual nature of the optimal portfolio mix.     

一株茶树叶部内生木霉的分离和鉴定

以健康茶树为材料,采用组织分离法分离获得1株内生真菌CSN-16。该真菌的孢子形状、菌丝形态、菌落生长速度等形态学特征与木霉的特征基本一致;进一步进行分子鉴定,获得该菌株的18S rDNA部分序列,并提交Genebank,获得基因登录号为EU678669。18S rDNA序列系统进化分析显示该菌株与拟康氏木霉(Trichoderma pseudokoningii)的同源性最高,相似性达99%。初步鉴定该菌株为拟康氏木霉。     

油茶种子水通道蛋白CoPIP1-1的鉴定与分析

以构建的油茶cDNA文库为基础,采用交错延伸PCR技术,分离克隆到一个水通道蛋白基因的编码序列(coding sequence,CDS),它编码一个287aa的跨膜蛋白(GenBank蛋白质id:ACF39901)。该蛋白可能由2条基因(GenBank登录号:EU850810、EU850811)编码,这2个基因具有相同的CDS和3′-UTR,但5′-UTR不同,其中之一多了2个小的插入片段。经对该蛋白的同源性比对和特征性基序分析,推测这个水通道蛋白属于质膜内在蛋白成员,定名为CoPIP1-1。通过同源建模,论证CoPIP1-1的水通道活性与通道口的(去)覆盖有关。N端与D环、B环的静电势作用导致了对通道口的(去)覆盖,并受磷酸化/质子化门控机制调控,质子化抑制通道活性,磷酸化则解除抑制。从同源建模的结果和细胞内的氧化状态推测该蛋白为组成型的低活性,这可能是油茶种子近成熟期脱水的成因之一。     

Identification and quantitative analysis of stage-specific carbohydrates in loblolly pine (Pinus taeda) zygotic embryo and female gametophyte tissues

Stage-specific analyses of starch and 18 sugars, including pentoses, hexoses, disaccharides, trisaccharides, oligosaccharides and sugar alcohols, were made throughout seed development for zygotic embryo and female gametophyte (FG) tissues of loblolly pine (Pinus taeda L.). Tissue was most often analyzed in triplicate from two open-pollinated families grown in different locations and sampled in different years. Carbohydrates were analyzed by enzymatic assay, high performance liquid chromatography or gas chromatography/mass spectrometry. For all carbohydrates quantified, peak concentrations were higher in embryo tissue than in FG tissue. Significant changes in starch and sugar concentrations occurred over time, with both seed collections showing similar trends in temporal changes. Although concentrations were not always similar, embryo and FG tissues generally showed similar patterns of change in starch and sugar concentrations over time. Total starch concentration was highest during early seed development and decreased as development progressed. The major sugars contributing to osmotic potential during early seed development were D-pinitol, sucrose, fructose and glucose. During mid-seed development, D-pinitol, sucrose, fructose, glucose, melibiose and raffinose provided major contributions to the osmotic environment. During late seed development, sucrose, raffinose, melibiose, stachyose and fructose were the major contributors to osmotic potential. These data suggest stage-specific media composition for each step in the somatic embryogenesis protocol.     

Regeneration of pitch pine (Pinus rigida) stands inhibited by fire suppression in Albany Pine Bush Preserve,New York

The Pine Barrens in New York State are dominated by pitch pine(Pinus ridgida) and scrub oaks(Quercus ilicifolia and Q. prinoides). With fire suppression over the last 90 years or so, P. rigida regeneration has become sparse, even in areas set aside to preserve this type of vegetation. We evaluated the effects of fire and an alternative disturbance(mechanical removal of vegetation)for increasing P. rigida regeneration in the Albany Pine Bush Preserve in eastern New York State. By comparing cover, diameter and height of P. rigida, Q. ilicifolia, Q.prinoides, and Vaccinium spp.(blueberry) growing on the sites with a known fire history, we documented structural and compositional changes in the vegetation since its establishment. We also compared sites on which Quercus sprouts were controlled through mechanical removal. In the years following a disturbance, height and cover of Q. ilicifolia increased continuously and the species became increasingly competitive. Both the density of P. rigida seedlings and Vaccinium cover decreased significantly as the diameter and height of Quercus increased. Our results confirmed that the dominance by P. rigida could be attributed to periodic fires at intervals of 20–25 years. We conclude that removing Q. ilicifolia can favor P. rigida dominance on the barren without fire by releasing pine seedlings from competition.     

Identification of the genus Armillaria by specific amplification of an rDNA-ITS fragment and evaluation of genetic variation within A. ostoyae by rDNA-RFLP and RAPD analysis

Genetic variation among Armillaria ostoyae isolates was studied by rDNA-restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) analysis. A total of 20 A. ostoyae isolates, mainly obtained from Picea spp. of different geographical origins, were examined. Southern hybridization of whole-cell DNAs digested with AvaII and probed with biotin-labelled cloned rDNA from Saccharomyces carlsbergensis allowed the differentiation of five RFLP groups. UPGMA cluster analysis of RAPD profiles (138 scorable bands) generated by 10 decamer primers (OPA 01-OPA 10) grouped the isolates in subclusters at similarity levels between 40% and 96%, indicating high intraspecific genetic variability. Some isolates of different geographical origins subgrouped together, suggesting that similar mutational events have occurred independently and that genetic exchange and recombination occurs among the DNAs in natural populations. The potential role of historical and current spread of spruce plants on the genetic variation of A. ostoyae isolates in Europe is discussed. Using the primer pair ARM-1 and ARM-2, an Armillaria-specific ITS-DNA fragment of about 660 bp was obtained. No intraspecific RFLP of this amplicon could be revealed, indicating low genetic variability of this region. The established informative RFLP and RAPD markers and also the Armillaria-specific ITS-DNA fragment may be powerful tools for further epidemiological, phylogenetic and host-pathogen interaction studies with A. ostoyae.     

核桃根系分泌物化感物质的分离与鉴定

为了探讨核桃根系分泌物的化感作用,利用乙酸乙酯提取核桃活体收集到的根系分泌物并通过气相色谱和质谱联用技术(GC-MS)进行分离鉴定,研究根系分泌物对萝卜种子发芽、胚芽和胚根伸长的影响.结果表明:根系分泌物酸性、中碱性组分对萝卜种子发芽、胚芽和胚根伸长都有抑制作用;核桃根系分泌物酸性组分包括烷烃、烯烃、酸、酯、醇、酚、酮、胺、醌、苯等;中性组分包括烷烃、烯烃、酸、酯、醇、酚、萜、胺、萘、醌、苯等;碱性组分包括烷烃、烯烃、酸、醇、酚、胺、醌、酮、苯等.     

Improvement in growth and salt resistance of lemon (Citrus limon) trees by an interstock-induced mechanism

Interstocks can reduce toxic ion accumulations in leaves of budded citrus trees, but the mechanism is not understood. We grew sour orange (Citrus aurantium L.; SO) seedlings, budded trees of 'Salustiano' orange (Citrus sinensis L. Osbeck; SAO) on SO, 'Verna' lemon (Citrus limon L. Burm. f; VL/SO) and interstock trees (VL/SAO/SO) in pots of sand watered with nutrient solution containing 5 (control) or 50 mM NaCl (saline treatment) for 12 weeks. Plants were harvested in six successive harvests and time trends in relative growth rate (RGR) and its components were estimated by fitting a Richards function regression to the harvest data. The VL/SAO/SO trees in saline conditions had higher mean RGR than VL/SO trees in control conditions. Increases in both net assimilation rate on a leaf mass basis (NARw) and leaf mass fraction (LMF) contributed equally to a twofold increase in RGR of VL/SAO/SO trees in saline conditions. In control conditions, the increase in RGR caused by the interstock had growth response coefficients of GRC(NARw) = 0.20 and GRC(LMF) = 0.80. Structural modifications-specific leaf area, leaf area ratio and LMF-had a slight influence on the salt-induced changes in RGR, whereas NARw had a large influence. Salinity decreased root mass fraction (RMF) and increased stem mass fraction (SMF). In contrast, the interstock decreased SMF and increased LMF and RMF. The VL/SAO/SO trees had the highest RMF and proportionally higher Cl- and Na+ allocations in roots than the other plant types. In saline conditions, reductions in leaf ion transport rate and dilution of imported ions by foliar growth nearly halved ion accumulations in leaves of VL/SAO/SO trees.     

红外光谱和元素分析对竹材炭化机理的研究

通过测定在200-600℃炭化竹材得到的固体产物的碳、氢、氧元素的含量及它们的红外光谱,研究了在炭化过程中竹材中半纤维素、纤维素及木素的变化规律。结果表明,结合元素分析,红外光谱分析方法是研究竹材炭化机理的有效手段。在200℃以前,竹材中的半纤维素和纤维素的大量羟基断裂,并结合成水而失去。在200-250℃之间,竹材中的纤维素被降解,其中的吡喃型环也遭到破坏。并且木素中的甲氧基也被脱去。竹材中的木素网状结构在250-400℃之间遭到完全的破坏。竹炭中的碳原子在600℃已基本上完成了芳环化。图3表2参15。     

常山胡柚叶片DNA的抽提与分析

经反复实验，采用在抽提前加入一定量的抗氧化剂β－巯基乙醇，用冰冻的异丙醇来沉淀DNA，并增加氯仿／异戊醇抽提次数的所谓改良CTAB法，对常山胡柚叶片DNA抽提与分析，不仅成功提取到常山胡柚叶片DNA，而且所得的DNA浓度高、纯度好，20个试样的DNA苹均质量浓度为272.86μg/mL，平均每克鲜叶片可获DNA约82μg，纯度基本上都在1.4-1.8范围内，并经RAPD反应验证，完全能满足常规分子生物学分析的要求。     

桃水孔蛋白基因克隆及序列分析

为给水孔蛋白(AQP)在低温对花芽发育和休眠调控中的作用研究奠定基础,以低温休眠的桃"满天红"花芽为试材,提取总RNA,根据其它植物AQP基因保守区设计简并引物,采用RT-PCR方法,扩增出3个编码长约147个氨基酸序列的水孔蛋白基因PpPIP1-1、PpPIP1-2和PpPIP2-1。同源性比对结果表明这些序列与其它物种AQP基因具有较高的同源性。这3个AQP基因的氨基酸序列均具有MIP家族的特征信号序列SGXHXNPAVT,还具有植物质膜水孔蛋白(PIP)家族的特征信号序列GGGANXXXXGY和TGI/TNPARSL/FGAAI/VI/VF/YN。经系统发育分析将这3个AQP基因分为2类,分属PIP蛋白的PIP1和PIP2亚家族成员。