白桦雌花序cDNA消减文库的构建及分析

为研究白桦雌花序花期基因表达情况,以两天为间隔对其进行取样.基于SMART策略,通过RT-PCR,将源自最后时期样品的cDNA作为Driver cDNA,源自其他时期样品的cDNA作为Tester cDNA,构建抑制性消减文库.EST序列经blastX分析表明,该文库中的基因大致可以归为五类,分别同代谢、物质运输和信号转导、细胞周期、胁迫反应及调控相关.本文对基因表达同发育的关系做了探讨.     

平榛ChWRKY28基因克隆及表达模式分析

[目的]研究平榛ChWRKY28基因序列特征及其在不同非生物胁迫下的表达规律.[方法]以平榛为试材,采用RACE-PCR方法进行基因克隆;利用实时荧光定量PCR方法检测基因在不同组织及不同非生物胁迫下的表达模式.[结果]表明:克隆得到的WRKY基因,全长1 342 bp,基因内部包含1个长963 bp的完整开放阅读框,编码320个氨基酸残基,命名为ChWRKY28.构建的系统发育树表明:该序列与拟南芥AtWRKY28及杨树PtrWRKY93的关系最近,相似性分别为49%和60%.基因表达分析表明:ChWRKY28在雄花序、雌花芽及茎中均有表达,但在茎部(皮)中的表达量高于雄花序和雌花芽中的表达量,具有组织表达特异性;低温、干旱及盐胁迫均能诱导ChWRKY28基因的表达,但受诱导程度存在差异.亚细胞定位分析结果表明:ChWRKY28蛋白分布在细胞核内,是一个核蛋白.[结论]推测ChWRKY28基因可能参与植物响应非生物胁迫的信号转导过程.     

Cymolomia hartigiana (Saxesen, 1840) (Lepidoptera: Tortricidae) attacking toAbies nephrolepis max. new to Daxing'anling, Heilongjiang, China

An Olethreutine speciesCymolomia hartigiana (Saxesen, 1840), attacking toAbies nephrolepis Max., is reported for the first time from Daxing'anling, Heilongjiang, China. The morphological characteristics are briefly re-described with illustrations. Also its damage and biology are noted. Foundation item: This study was carried out under the financial support by KOSEF (Korea Science & Engineering Foundation) with the program of “Korea and China Young Scientist Exchange Program” (2002–2003). Biography: *BYUN Bong-Kya (1963-), male, Ph.D., Researcher in Korea National Arboretum, Korea. Responsible editor: Chai Ruihai     

Notes on genus Eurydoxa Filipjev （Lepidoptera： Tortricidae） in China

Genus Eurydoxa Filipjev in China is reviewed and noted for the first time. Based on the present study, two species are recognized, including rhodopa Diakonoff and advena Filipjev. All available information for the species is reviewed and provided.     

松材线虫Bxpel2基因的克隆及RNA干扰载体构建

[目的]为了构建松材线虫(Bursaphelenchus xylophilus)果胶酶Bxpel2基因干扰载体.[方法]通过Trizol 法提取松材线虫总RNA,反转录合成cDNA,设计带T₇启动子的果胶酶Bxpel2基因引物,以cDNA为模板扩增出果胶酶Bxpel2基因片段,连接到RNA干扰载体,再以干扰载体为模板,PCR扩增出目的片段后进行测序鉴定,合成果胶酶Bxpel2基因双链RNA(dsRNA),采用RT-PCR检测松材线虫Bxpel2基因干扰后的表达情况.[结果]表明:1)提取的松材线虫总RNA完整性好,无降解;2)成功克隆出松材线虫果胶酶Bxpel2基因片段(790 bp)并将其连接至pMD19-T载体;3)以RNA干扰载体为模板合成dsRNA,浓度分别为1.313 mg·mL^-1和1.152 mg·mL^-1;4)RT-PCR结果显示,松材线虫经过dsRNA干扰后,Bxpel2基因表达基本受到抑制.[结论]成功构建松材线虫果胶酶Bxpel2基因干扰载体,为进一步研究Bxpel2基因在松材线虫致病过程中的作用和功能奠定了良好的基础.     

Nitrate reductase activity and its diurnal variation rhythm for Camptotheca acuminata seedlings

Nitrate reductase activity (NRA) in different plant organs and leaves in different positions of Camptotheca acuminata seedlings was determined by an In vivo assay, the diurnal variation rhythm of NRA in leaves of different positions was observed,and the correlations between leaf NRA, leaf area and lamina mass per unit area (LMA) were also examined. The results showed that NRA in the leaf was significantly highest, compared with that in other organs such as roots, stems and leaves. In this experiment, the 10 leaves were selected from the apex to the base of the seedlings in order. The different NRA occurred obviously in leaves of different positions of C. acuminata seedlings from the apex to the base, and NRA was higher in the 4th-6th leaves.The diurnal change rhythm of leaf NRA showed a one peak curve, and maximum NRA value appeared at about midday (at 12:30 or so). No obvious correlations between NRA and leaf area or lamina mass per unit area were observed. This study offered scientific foundation for the further research on nitrogen metabolism of C. acuminata.     

寄生小蜂对小青×黑杨树皮及杨圆蚧挥发油的行为反应

Four-armed airflow olfactometer was used to determining the foraging behavior ofPteroptrix longgiclava (Girault) (Hymenoptera: Aphelinidae) andEncarsia gigas (Tshumakova) (Hymenoptera: Aphelinidae) to the essential oils which emitted from the healthy bark ofPopulus pseudo-simonii×P. nigra, the infested bark injured byQuadraspidiotus gigas (Thiem & Gerneck), the body and scale of fixed 1st-instar-nymph ofQ. gigas. The results from these experiments showed that the volatile oils produced from the injured bark and from the scale of fixed 1st-instar-nymph had a higher attractive ability to female adults of the two species of wasps. The essential oil produced from the scale of the pest at dosages of 3–7 μL and the essential oil emitted by injured bark at dosages of 5–9 μL had a stronger alluring effect on the host searching behavior ofPteroptrix longgiclava. The essential oil from the body of fixed 1st-instar-nymph ofQ. gigas also had certain effect on the host locating effort ofPteroptrix longgiclava andEncarsia gigas. Those two wasps did not shown any reaction to the essential oil produced by the healthy bark of poplar. Foundation item: This paper was supported by National Natural Science Foundation of China (39970620) and the “TRAPOYT” Biography: CHI De-fu (1962-), male, Ph.D., Professor in Northeast Forestry University, Harbin 150040, P.R. China Responsible editor: Chai Ruihai     

高效液相色谱法测定羟基喜树碱含量方法的研究

The determination method of 10-hydroxycamptothecin inCamptotheca acuminatu fruits by high-performance liquid chromatogram (HPLC) was studied. The HPLC analysis was performed on a HIQ sil C₁₈(4.6×250 mm) column with mobile phase of acetonitrile-water (3:7, V:V), flow rate 1 mL·min⁻¹ and UV detective wavelength 266 nm. Extracting 10-hydroxycamptothecin by ultrasonic method from fruits ofC. acuminata to prepare samples for analysis was systematically discussed. The optimal extraction condition was carried out by 60% alcohol solution at 60°C for 50 minutes. Foundation Item: This paper was supported by National Natural Science Foundation of China (No. 30070086). Biography: MA Mei-fang (1976-), female, Graduate Student of Northeast Forest University. Responsible editor: Song Funan     

Responses of fractal dimensions ofPicea koraiensis seedlings to different light environments

The changes of fractal dimension ofPicea koraiensis seedlings under different light intensities in natural secondary forests was studied. The results showed that with the change of light environment, crown characters ofPicea koraiensis seedlings exhibited a greater plastic in lateral number, lateral increment, lateral dry weight, and specific leaf area. The range of calculated fractal dimensions of seedling crowns was confined between 2.5728 and 2.1036, but maximum of fractal dimension achieved in term moderate shading and in extreme low light conditions fractal dimension was least. Foundation item: This paper was supported by the National Natural Science Foundation of China (No.39670152) and Chinese Academy of Sciences. Biography: ZHOU Yong-bin (1970-), female, Ph. Doctor, lecture in Shenyang Agricultural University, Shenyang 110161, P.R. China Responsible editor: Song Funan     

Protein diversity ofPaulownia plant leaves and clusters

Paulownia tomentosa, P. fargesii, P. lamprorhylla, P. albiphloea, P. australis, P. fortunei, P. elongata, P. elongata f. alba andP. albiphloea varchenggtuensis were classified into three groups:P. fortunei group (P. fortunei andP. elongata f. alba);P. australis group (P. australis andP. albiphloea varchenggtuensis) andP. tomentasa group (P. tomentasa, P. fargesii, P. albiprhlaca, P. lamproprhylia andP. elongata) accordance to the results of the single and two-dimensional SDS-PAGE of protein in thePaulownia tree leaves. The result could lay a foundation for classifying the GenusPaulownia plants. Foundation Item: This paper was supported by National Nature Science Foundation of China and Nature Science Foundation of Henan Province. Biography: FAN Guo-qiang (1964-), male, Professor in Institute ofPaulownia Henan Agriculture University. Zhengzhou 450002, P. R. China. Responsible editor: Song Funan     

哈茨木霉天冬氨酸蛋白酶基因SA76的3＇RACE扩增和序列分析

由EST获得全长cDNA对于结构基因组学和功能基因组学都是至关重要的,cDNA末端快速扩增技术RACE是该领域中的重要研究方法.利用BD SMART RACE技术扩增编码分泌天冬氨酸蛋白酶SA76基因的3＇末端,将其与哈茨木霉cDNA文库中的SA76基因的EST序列进行序列拼接,获得2019bp的全长cDNA序列,其开放读码框长1593bp,5＇非编码区266bp,3＇非编码区201bp,编码530个氨基酸,有信号肽.哈茨木霉天冬氨酸蛋白酶基因与玉蜀黍赤霉、粗糙脉孢菌、球毛壳菌天冬氨酸蛋白酶基因的同源性分别为53%, 37%, 36%.利用BD SMART RACE技术首次从哈茨木霉中克隆天冬氨酸蛋白酶基因,为验证SA76基因的功能奠定基础,为进一步研究蛋白酶的作用机制及生物防治功能提供依据.     

中间锦鸡儿CiDR1的克隆及干旱胁迫下的表达分析

[目的]研究并了解中间锦鸡儿CiDR1基因功能及其对干旱胁迫的响应,为抗性育种提供候选基因。[方法]通过RACE技术从中间锦鸡儿中克隆CiDR1基因的c DNA全长,利用生物信息学分析软件对其基因结构及功能进行分析和预测。再通过qRT-PCR技术对干旱胁迫后的幼苗中的CiDR1表达模式进行研究。[结果]从中间锦鸡儿中克隆到CiDR1基因的c DNA全长共计4 297 bp,Gen Bank登录号为KP277100。生物信息学分析表明,预测的CiDR1蛋白序列中含有1 243个氨基酸残基,具有抗病基因特征结构域TIR、NB-ARC、LRR等,其等电点为6.35,不稳定指数为42.91,不具备信号肽,为非分泌蛋白,定位于细胞质中。定量PCR检测发现,CiDR1基因在幼年期的茎中表达量较低,在成年期的叶片中表达量较高;在干旱胁迫处理后的幼苗中,CiDR1表达水平有明显下降,表明该基因的表达受干旱抑制,可能与中间锦鸡儿适应干旱相关。[结论]中间锦鸡儿在干旱胁迫后其根、茎和叶中CiDR1的表达均明显下降,表明CiDR1的表达受干旱抑制,可能与中间锦鸡儿适应干旱相关,进一步研究发现CiDR1在根、茎、叶中的表达水平可能受发育阶段调控。     

Study on the biological characteristics ofHomalotglus flaminus

Homalotglus flaminus (Daiman) is an important natural enemy of theChilocorus kuwanae. Its biological characteristic, life history, the action style on hosts were studied at Hongqi Forest Farm, Daqing City, Heilongjiang Province from June 1998 to October 2000, in order to controlH. flaminus and raise the control ability ofChilocorus kuwanae toQuadraspidictus gigas pest. The observed results indicated thatH. flaminus has two generation one year in study area, the pupating span is 12.7 d, life span of females and males is 17.6 d and 10.8 d respectively, egg span is 10 d, and the sex ratio of females to males is 1∶3 Foundation item: This paper was supported by Natural Science and Foundation of Heilongjiang Province. Biography: Mpa Ling (1963), female, Ph. Doctor, associate professor in Northeast Forestry University, Harbin 150040, P. R. China Responsible editor: Song Funan     

Control of Lymantria dispar L. by biological agents

The experiment on control ofLymantria dispar L by using different kinds of biological measures, including nuclear polyhedrosis virus (NPV) ofLymantria dispar L., BtMP-342, sex-attractant as well as botanical insecticide, was carried out in the forest regions of Inner Mongolia in 2003. Two concentrations (2.632×10⁶ PIB·ml⁻¹ and 2.632×10⁷ PIB·ml⁻¹) ofLymantria dispar L. NPV were sprayed on the 2rd-instar-larvae ofL. dispar and 70% and 77.8% control effect were obtained respectively. BtMP0-342 was applied to the 3rd- and 4th-instar larvae and the control effect was around 80%. The sex-attractant provided by Canada Pacific Forestry Research Center also showed a good result in trappingL. dispar adults. The self-produced botanical insecticide, which was extracted from a kind of poisonous plant distributed in Daxing'an Mountains, China, exhibited a good control result in controlling the larvae ofL. dispar, and 82% mortality was observed when spraying primary liquid of the botanical insecticide on the 3rd–5th-instar-larvae in lab. Foundation item: This paper was supported by Natural Science Foundation of Heilongjiang Province (C0235) and Youth Foundation of Harbin City (0061218024). Biography: ZHANG Guo-cai (1964), male, Ph. Doctor Candidate, associate professor in School of Forestry, Northeast Forestry University, Harbin 150040, P. R. China. Responsible editor: Song Funan     

Genetic variation and division of Pinus sylvestris provenances by ISSR markers

Inter Simple Sequence Repeat (ISSR) was used to detect genetic variation among nine provenances, including 135 individuals. A total of 108 loci were amplified using 10 random primers. The differentiation of the percentage of polymorphic bands (PPB) among different provenances was evident, ranging from 27% to 54%, of which Honghuaerji provenance had the highest PPB and Kalunshan provenance had the lowest PPB. Shannon's Information index (I) at species level was 0.1581 and Nei's gene diversity (h) was 0.2393. Coefficient of gene differentiation (G_ST) calculated by Popgene was 0.3965, these results indicated that majority of genetic variation (60.35%) was found within provenances. According to dendrogram amongpinus sylvestriv provenances, nine provenances were divided into two provenance areas, namely Daxing's and Xiaoxing'an Mountains provenance area and Hulunbeier provenance area. Foundation item: This study was supported by National Scientific and Technological Brainstorm Project. Biography: LI Hui-yu (1978-), female, Ph.D. candidate, Northeast Forestry University, Harbin 150040, P. R. China. Responsible editor: Song Funan     

大花蕙兰转齿兰环斑病毒外壳蛋白基因及检测

[目的]通过植物转基因技术获得抗病毒大花蕙兰种质资源,优化转化体系和鉴定方法.[方法]本研究克隆了齿兰环斑病毒外壳蛋白基因,并构建了该基因的pBI121表达载体,用根癌农杆菌介导法转化大花蕙兰,尝试以巢式PCR方法检测转基因再生植株.[结果]优化了大花蕙兰遗传转化体系,建立了利用巢式PCR技术检测转基因大花蕙兰植株的方法,获得了32株转基因株(系).[结论]优化了以类原球茎为外植体的农杆菌介导转化大花蕙兰的方法,确定以5%~10%类原球茎存活时的抗生素(卡那霉素)浓度为筛选浓度;获得了转ORSV CP基因大花蕙兰植株;对大花蕙兰转基因植株检测时,巢式PCR较普通PCR更灵敏、准确.     

Cloning and sequence analysis of a glucose-6-phosphate dehydrogenase gene <Emphasis Type="Italic">PsG6PDH</Emphasis> from freezing-tolerant <Emphasis Type="Italic">Populus suaveolens</Emphasis>

A 1207 bp cDNA fragment (PsG6PDH) was amplified by RT-PCR from cold-induced total RNA of the freezing-tolerant P. Suaveolens, using primers based on the highly conserved region of published plant glucose-6-phosphate dehydrogenase (G6PDH) genes. The sequence analysis showed that PsG6PDH coding region had 1 101 bp and encoded 367 predicted amino acid residues. Moreover, the nucleotide sequence of PsG6PDH showed 83%, 82%, 79%, 79% and 78% identity, and the derived amino acid sequence shared 44.2%, 44.7%, 42.0%, 40.5% and 43.9% identity with those of the Solanum tuberosum, Nicotiana tabacum, Triticum aestivum, Oryza sativa and Arabidopsis thaliana, respectively. The results show that PsG6PDH is a new member of G6PDH gene family and belongs to the cytosolic G6PDH gene. This is the first report on cloning of the G6PDH gene from woody plants.     

A preliminary analysis of phylogenetic relationships ofArundinaria and related genera based on nucleotide sequences of nrDNA (ITS region) and cpDNA (trnL-F intergenic spacer)

Phylogenetic relationships ofArundinaria and related genera (Pleioblastus, Pseudosasa, Oligostachyum, Bashania, Clavinodum, etc.) were assessed by analyzing the sequences of the nrDNA internal transcribed spacer (ITS) and the cpDNAtrnL-F intergenic spacer (IGS). Comparison withtrnL-F IGS sequence, the ITS region provided the higher number of parsimony informative characters, and the interspecific variation of the ITS sequence was higher than that of thetrnL-FIGS sequence. The tree obtained by combining both sets of data showed that the species sampled inArundinaria and the related genera were monophyletic and divided into two clades. The relationships and positioning of all the taxa surveryed (includingA. oleosa, A. hsienchuensis, A. chino, A. amara, A. yixingensis, A. amabilis, A. fortunei, A. pygmaea, A. gramineus, A. fargesii, A. faberi, A. hupehense, Pseudosasa japonica cv. Tsutsumiana, P. japonica andBrachystachyum densiflorum) were also discussed. The results from the sequences were broadly consistent with morphological characters, appearing all these taxa sampled belong to the genus ofArundinaria. The topologies of the trees generated from individual data and the combined data were similar. Foundation item: This study was supported by National Natural Science Foundation of China (No. 30170788) Biography: ZHUGE Qiang (1959-), male, professor in Laboratory of Forest Genetics and Gene Engineering, Nanjing Forestry University, Nanjing 210037, P.R. China. Responsible editor: Song Funan     

Research on the extension ofTamarix shrubs resulted from development projects in arid area

Tamarix plant is widely distributed in the arid areas of China for its great tolerance of draught, salt and alkali and attracts more attention by its strong capacity in windbreak and sand-fixations. “Tamarix dunes” constitute special desert bio-landscape in arid area. During last several decades,Tamarix shrubs declined and “Tamarix dunes” were also severely destroyed due to the land exploitation and serious deforestation, etc. From the 1980s until now, the extending phenomenon ofTamarix shrubs caused by developmental projects such as building highway, railway was studied in the western dry area of China, including western section of Hexi Corridor of Gansu, railroads and highways around the two greatest basin in Xinjiang and interior regions in Gurbantonggut desert, through on-the-spot survey along the transportation lines and setting up sampling plots along the roads. It was found that large quantities ofTamarix plants grow in the catchment area of low-lying lands that were formed by bulldozer operation during road building period. The extension ofTamarix shrubs caused by engineering was similar to that of the other section of the area to some extent, but went beyond the original distribution. This extension is beneficial to the ecological restoration and re-vegetation of western region of China and plays an important role in control of the sandstorms and improvement of ecological environments. As a result, it is necessary to make a further study on the extending phenomenon ofTamarix shrubs and to seek approach to promote wider extension ofTamarix shrubs in suitable habitats. Foundation Item: This study is supported by key projects of National Natural Science Foundation (39730100), National Foundation (30070640) and part of Project 973(G19990435). Biography: Huang Pei-You (1936-), Male, Professor in College of Life Science and Technology, Xinjiang University, Urumqi, 830046 Responsible editor: Chai Ruihai     

Bio-control trials of Chaetomium spirale ND35 against apple canker

A new endophytic antagonistic fungus, Chaetomium spirale ND35 from Populus tomentosa, was reported. The bio-control trials of C. spirale ND35 against the Valsa Canker of apple were preliminarily investigated. The results of dual culture on PDA plate showed that C. spirale ND35 was capable of strong antagonism against Valsa ceratosperma, and for inhibiting the mycelial growth of V. ceratosperma,.the crude extract of liquid culture of corn steep powder broth was more effective than that one of malt extract broth (MEB). The results of bio-control in greenhouse and field indicated that the disease incidence of apple tree treated with C. spirale ND35 was lower significantly than that treated by other methods. The re-isolation experiment suggested that C. spirale ND35 could colonize in stems and branches of apple trees successfully, and the ND35 colonization rate of the treatment with solid wheat bran culture was higher than that of corn steep powder broth, but the field experiment result the control effect of liquid culture of C. spirale ND35 was better than that of solid culture.