We investigated the inheritance and expression of cry1A gene in transgenic maize (Zea mays L.) by Southern blotting analysis and enzyme-linked immunosorbent assays (ELISA). The results showed that cry1A had been transmitted to progeny of transgenic maize as a single gene. Contents of cry1A insecticidal protein were significantly different among transgenic maize lines and various tissues of the same transgenic lines. High expression of cry1A protein occurred in green tissues, such as leaf and husk leaf, and low expression occurred in pith, tassel, ear pith, pollen and silk. The results also showed that the contents of cry1A insecticidal protein in leaves of transgenic maize increased with the advance of development and there was no significant difference in cry1A expression level among various generations of transgenic maize. Key words: cry1A gene; transgenic maize; inheritance; expression; insecticidal protein
通讯作者。Tel:010-62892577; E-mail:gywang@ mail.cau.edu.cn。
For finding valuable systematic characters, leaf epidermis of 77 taxa, representing 12 sections of the genus Clematis and three related genera in the Ranunculaceae, were examined mainly by means of light microscopy (LM), and partially by scanning electron microscopy (SEM). It was shown that the leaf epidermal cells were usually irregular or polygonal in shape. The patterns of anticlinal walls were straight, arched or undulate. The stomatal apparatus is anomocytic and exists in abaxial epidermis of all species, and in the adaxial epidermis of some species. Under SEM observation, the leaf epidermis was often striated, seldom nearly smooth, and often with flakes attached. Evidence from leaf epidermis serves as a criterion for distinguishing the subsections in sect. Meclatis(Spach)Tamura and in sect. Fruticella Tamura. The results also support that there are several separate evolutionary processes in the genus Clematis. Key words: Ranunculaceae; Clematis; leaf epidermis; stomatal apparatus
摘要： 用光学显微镜和扫描电镜,较全面地观察了铁线莲属(Clematis)植物和邻近类群的叶表皮特征,并对其系统学意义进行了讨论.在光学显微镜下,观察了铁线莲属12组63种5变种及其同族邻近类群3属9种;在扫描电镜下,观察了铁线莲属7组27种2变种及其邻近类群2属6种植物的成熟叶表皮细胞及气孔器的特征.在光学显微镜下可见:叶表皮细胞形状(表面观)为多边形、不规则形,垂周壁式样为较平直、浅波状和波状,气孔器普遍存在于下表皮,少数种的上表皮也有分布,其类型均为无规则型.在扫描电镜下可见:叶片通常具有条纹或有颗粒状或鳞片状附属物,气孔器外拱盖内缘通常为浅波状或近平滑.在黄花铁线莲组(sect. Meclatis (Spach) Tamura)和灌木铁线莲组(sect. Fruticella Tamura)中,叶表皮具有双面气孔器的特征对于组下亚组的划分有一定意义.并且叶表皮特征对于一些种的分类学处理也有一定的价值,同时也为属下组间的系统进化关系提供了一些线索.
通讯作者。E-mail: LqLi @ ns.ibcas.ac.cn.。
Study on seasonal responses of terrestrial net primary production (NPP) to climate changes is to help understand feedback between climate systems and terrestrial ecosystems and mechanisms of increased NPP in the northern middle and high latitudes. In this study, time series dataset of normalized difference vegetation index (NDVI) and corresponding ground-based information on vegetation,climate, soil, and solar radiation, together with an ecological process model, were used to explore the seasonal trends of terrestrial NPP and their geographical differences in China from 1982 to 1999. As the results, seasonal total NPP in China showed a significant increase for all four seasons (spring,summer, autumn and winter) during the past 18 years. The spring NPP indicated the largest increase rate, while the summer NPP was with the largest increase in magnitude. The response of NPP to climate changes varied with different vegetation types. The increased NPP was primarily led by an advanced growing season for broadleaf evergreen forest, needle-leaf evergreen forest, and needle-leaf deciduous forest, whilst that was mainly due to enhanced vegetation activity (amplitude of growth cycle) during growing season for broadleaf deciduous forest, broadleaf and needle-leaf mixed forest, broadleaf trees with groundcover, perennial grasslands, broadleaf shrubs with grasslands, tundra, desert, and cultivation. The regions with the largest increase in spring NPP appeared mainly in eastern China, while the areas with the largest increase in summer NPP occurred in most parts of Northwestern China, Qinghai-Xizang Plateau, Mts. Xiaoxinganling-Changbaishan, Sanjiang Plain, Songliao Plain, Sichuan Basin, Leizhou Peninsula, part of the middle and lower Yangtze River, and southeastern mountainous areas of China. In autumn, the largest NPP increase appeared in Yunnan Plateau-Eastern Xizang and the areas around Hulun Lake. Such different ways of the NPP responses depended on regional climate attributes and their changes. Key words: Carnegie-Ames-Stanford-Approach (CASA) model; net primary production (NPP); seasonal change; normalized difference vegetation index (NDVI); climate change
通讯作者。E-mail: jyfang @ urban.pku.edu.cn。
通讯作者。D/G%"!：h JBA%,-i7(&%,; ?]7; >H7; ), j 。
To understand the dynamics of added nitrogen (N) in alpine meadow and the role of alpine plants and soil microorganisms in the retention of deposited N, the fate of 15N labeled nitrate and ammonium salts was determined in an alpine meadow for two months. Two weeks after 15N application, total recovery of 15N from NO3--15N was 73.5% while it was 78% from NH4+-15N. More 15N was recovered in plants than in soil organic matter or in microbial biomass, irrespective of forms of N added. After one month, 70.6% of added NO3--15N and 57.4% of NH4+-15N were recovered in soils and plants. 15N recovered in soil organic matter decreased greatly while that recovered in plants varied little, irrespective of the form N. Compared with the results of two weeks after 15N application, more NO3--15N than NH4+-15N was recovered in microbial biomass. Total recovery was 58.4% (six weeks) and 67% (eight weeks) from NO3--15N 15N, and 43.1% and 49% from NH4+-15N, respectively. Both plants and soil microorganism recovered more NO3--15N than NH4+-15N. But plants recovered more 15N than soil microorganisms. During the whole experiment plants retained more NO3--15N and 15N than soil microorganisms while 15N recovered in inorganic N pool did not exceed 1% due to lower amount of inorganic N. This indicates that plants play more important roles in the retention of deposited N although microbial biomass can be an important sink for deposited N in early days after N application. Key words: alpine meadow; soil microorganisms; retention of deposited N; percentage of 15N recovery; fate of NO3--15N and NH4+-15N
徐兴良 欧阳华 裴志永 周才平
摘要： 运用15N稳定性同位素示踪技术,对高寒草甸植物和土壤微生物固持沉降氮的能力及沉降氮在小嵩草(Kobresia pygaea)草甸中的运移规律进行了研究.施肥2周后,NO3-15N和NH+4-15N的总恢复率分别为73.5%和78%.无论是NO3-15N,还是NH4+-15N,植物所固持的15N总是比土壤有机质或者是土壤微生物固持的多.4周后,70.6%的NO3-15N和57.4%的NH4+-15N被固持在土壤和植物中.其中,土壤有机质所固持的15N均下降了很多,而植物所固持的15N却变化很小.同前面的结果相比,较多的NO3-15N为土壤微生物所固持.在施肥6周和8周后,NO3-15N的总恢复率分别为58.4%和67%,而NH4+-15N的总恢复率分别为43.1%和49%.植物和土壤微生物所固持的NO3-15N比NH4+-15N多.在整个实验期间,植物固持的NO3-N较多,而且比土壤微生物固持了较多的15N.由于无机氮的含量一直很低,无机氮库所固持的15N一般不超过1%.上述结果意味着短期内植物在高寒草甸中对沉降氮的去向起着决定作用.
关键词： 高寒草甸；土壤微生物；15N恢复率；沉降氮的固持；NO 3- -N和NH4+-N运移规律
通讯作者。Tel: 010-64889697; E-mail: ohua @ igsnrr.ac.cn 。
Short-term batch cultures were used to measure the phosphate-dependent growth kinetics for a marine microalga, Tetraselmis subcordiformis (Wille) Hazen, and a marine macroalga, Ulva pertusa Kjellm. Results were fitted to the Monod model. U. pertusa had a lower half-saturation constant and maximum growth rate, which were 0.016 μmol/L and 0.16 d-1respectively，while the growth kinetics of T. subcordiformis were 0.021μmol/L and 0.83 d-1.Long-term semicontinuous nutrient competition experiments were performed between T. subcordiformis and U. pertusaunder phosphate limitation in laboratory. Loss rates were manipulated to get the same or different resource requirement values (R*) of the two species. Comparison between the theoretical predictions derived from Monod kinetics and the outcome of competition experiments indicated that the Monod model could predict the results only when the R* values of the two species were significantly different, and T. subcordiformis displaced U. pertusa when they had the same resource requirements. The Monod model can only partly predict th competition results between microalga and macroalga. Key words: macroalga; microalga; Monod model; phosphate; resource competition theory; R*-rule
南春容 双 林 金 秋
摘要： 用一次性培养法结合Monod方程测得海洋微藻-亚心型扁藻(Tetraselmis subcordiformis (Wile) Hazen)与大型海藻-孔石莼(Ulva pertusa Kjellm.)磷限制下的生长动力参数.孔石莼具有较低的半饱和生长常数及最大生长率,其分别为0.016 μmol/L和0.16 d-1,而亚心型扁藻的半饱和生长常数和最大生长率分别是0.021 μmol/L 和0.83 d-1. 两种藻类间的营养竞争实验采用半连续培养法在磷限制条件下进行,实验过程中,分别对两者施予相同或不同的去除率,使两者享有相同或不同的资源需求值R*.由Monod方程所作的竞争预测与实验观察结果的比较显示:仅在两种藻类间的资源需求值R*差异显著(t检验,P＜0.01)时,Monod方程才能对竞争结果作出较为准确的预测;在两种藻类享有相同的资源需求值R*时,亚心型扁藻在竞争中取代孔石莼.Monod模型仅能部分预测大型海藻与海洋微藻间的竞争结果.
通讯作者。E-mail:nanchunrong @ eyou .com。
By using the liquid/liquid diffusion method at a suitable crystallization conditions, large single and dark brown crystals (the sides of the largest crystals were 0.20 mm×0.20 mm×0.07 mm and 0.18 mm×0.18 mm×0.05 mm, respectively) could be obtained from the solutions of nitrogenase CrFe protein and MnFe protein purified from a mutant UW3 of Azotobacter vinelandii Lipmann grown in Cr- or Mn-containing but NH3-free medium. The time of crystal formation, as well as the number,size, shape and quality of crystals obviously depended on the concentrations of PEG, MgCl2 and NaCl. The liquid/liquid diffusion method seems to benefit CrFe protein and MnFe protein for the growth of large single crystals for X-ray diffraction analysis. Key words: mutant UW3 of Azotobacter vinelandii; nitrogenase CrFe protein and MnFe protein; growth of large single crystals
吕玉兵 赵 颖 赵剑峰 代小虎 仓怀兴 王耀萍 黄巨富
（1. 中国科学院植物研究所光合作用与环境分子生理学重点实验室，北京100093；2. 中国科学院生物物理研究所，北京100101）
摘要： 用液／液扩散法，从分别含Cr和Mn的无氨培养基中生长的固氮菌（Azotobacter vinelandii Lipmann)突变种UW3中纯化出的CrFe蛋白和MnFe蛋白，在合适的结晶条件下生长出深棕色大单晶（最大晶体的尺寸分别为0．20mm×0．20mm×0．07mm和0．18mm×0．18mm×0．05mm）。PEG、MgCl2和NaCl的浓度对这两种蛋白的出晶时间、晶核数目、晶体大小和形状都有明显影响。结果表明，用此结晶法有利于CrFe蛋白和MnFe蛋白生长出可供X－射线衍射分析的大单晶。
Plant growth rate (GR), contents of free polyamines (fPAs) and bound polyamines (bPAs) and activities of some key enzymes involved in polyamine (PA) metabolism in the roots of two barley (Hordeum vulgare L.) cultivars differing in salt sensitivity were investigated with 0-300 mmol/L NaCl treatments. With 0-200 mmol/L NaCl treatments, activities of arginine decarboxylase (ADC) and transglutaminase (TGase) and PA oxidase (PAO) in the roots of barley seedlings all increased, while TGase and PAO activities decreased slightly at 300 mmol/L NaCl. As a result, free Put (fPut) content increased continuously with increasing concentrations of NaCl, while levels of free Spd (fSpd) and an unknown PA (fPAx) and bPAs (bPut, bSpd and bPAx), as well as (fSpd+fPAx)/fPut ratio rose at 50-200 mmol/L NaCl and reduced at 300 mmol/L NaCl. However, no significant change in the tetra-amine spermine (Spm) content was observed. Statistical analysis showed that GR was very significantly positively correlated with (fSpd+fPAx)/fPut ratios and the contents of bPAs, whereas a significant inverse correlation existed between GR and the ratios of fPA contents to bPA levels. These results showed that, under salt stress, the balance between fSpd, fPAx and fPut levels and an equipoise between fPA and bPA contents in roots were important to salt tolerance of barley seedlings. Key words: barley (Hordeum vulgare); polyamine (PA); metabolism; salt tolerance
赵福庚 孙 诚 刘友良 章文华
摘要： 研究了0－300mmol/L NaCl对大麦（Hordeum vulgare L.)幼苗生长速率，根系游离和结合态多胺含量以及多胺生物合成关键酶活性的影响。结果表明，在0－200mmol/L NaCl处理下精氨酸脱羧酶（ADC）、多胺氧化酶（PAO）以及转谷酰胺酶（TGase)活性明显提高，而在300mol/L NaCl处理下活性下降，与之对应，游离腐胺（Put)含量随处理盐浓度的提高一直呈上升趋势。亚精胺（Spd)和在根系内检测到的未知多胺（PAx）在低浓度盐处理时含量上升，随盐浓度的提高含量下降，盐处理前后精胺（Spm)含量变化不明显，低浓度盐处理时游离态（Spd+PAx)/Put上升，随盐浓度的提高比值明显下降，结合态Put,Spd和PAx含量以及结合态多胺总量均在低浓度盐处理时上升，随盐浓度的提高含量明显下降，统计分析显示，大麦相对生长速率与游离态（Spd+PAx）／Put和结合态多胺含量间均呈极显著正相关关系，与游离态多胺和结合态多胺的比值间均呈显著负相关关系，上述结果说明盐胁迫下大麦体内游离态Spd,PAx与Put以及结合态形式之间的平衡与大麦耐盐性关系密切，游离态Put向Spd,PAx以及结合态形式转化均有利于大麦耐盐性的提高。
The switch from C3 to CAM pathway was induced by water stress in a C3 /CAM intermediate plant Sedum spectabile Boreau. Typical CAM criteria were observed upon 15 d of withholding water. Leaf δ13C value (-‰) and water content showed a linear correlation fashion. Chlorophyll fluorescence parameters and antioxidative capacity were altered by water stress.ΦPSⅡ and qP were reduced by 50% and 34% of the control, respectively, while NPQ rose ca .180%. SOD activity and ability to scavenge DPPH· free radical went down but membrane permeability changed slightly. However, when an additional photooxidation by MV with high PPFD was carried out with leaf discs from watered (C3 mode) and drought plants (CAM mode), qP andΦPSⅡ in leaves at induced CAM mode stage continuously decreased to a very low level. High 1-qP value (0.86) and 1-qP/NPQ ratio (>1)indicated the presence of high reduction state and unbalance of light energy budget. Together with the marked loss of membrane integral, it was evidenced that photooxidative damage was more serious in the induced CAM mode than in the C3 mode. No advantage of photooxidation tolerance was found at the induced CAM expression stage of the facultative CAM plant, as compared with its C3 mode stage, and also with the constitutive CAM plants reported previously. The differences in photooxidation sensitivity between the inducible CAM expressing plant and the constitutive CAM plant were discussed. Key words: Sedum spectabile; facultative CAM plant; photooxidation; water stress; CAM expression
兼性CAM 植物长叶景天叶片在!C3和CAM 型时的光氧化作用
林植芳 彭长连 林桂珠
摘要： 通过停止浇水产生水分胁迫使兼性CAM植物长叶景天（Sedum spectabile Boreau)叶片光合途径由C3型转为CAM型。干旱15d时观察到典型的CAM生理特征，且叶片的？13C值与含水量成线性相关。水分胁迫改变了叶绿素荧光参数和抗氧化能力，和ΦPSⅡ和qp降低34％，NPQ提高约180％，SOD活性和清除DPPH自由基能力也明显下降，但膜半透性变化不大，当将处于C3（浇水）和诱导为CAM（缺水）型的叶圆片用源甲基紫精（MV）和强光作光氧化处理后，与C3型叶片相比，诱导CAM型叶片的NPQ不能提高，qp和ΦPSⅡ降至很低水平，光系统处于高还原态，光能供给与消耗失衡（1－qp=0.86和（1－qp)/NPQ>1)，膜系统几乎失去完整性，这种严重的光氧化损伤表明，与我们以前报告的专性CAM植物诱导表达的CAM型未能显示比C3型较强的耐光氧化优势。讨论了出现这种光氧化敏感性差别的可能原因。 (共6页)
The protective role of xanthophyll cycle in resurrection angiosperm Boea hygrometrica (Bunge) R.Br. was investigated by analysis of the changes of chlorophyll fluorescence and xanthophyll cycle components in response to dehydration and rehydration in detached leaves under very weak light condition (3 μmol photons·m-2·s-1) and in the dark. With declines in the values of PSⅡ photochemical efficiency (Fv/Fm), PSⅡ actual quantum yield (ΦPSⅡ), photochemical quenching (qP) and non-photochemical quenching (NPQ) during dehydration, zeaxanthin significantly increased in control Boea leaves under very weak light condition, while no zeaxanthin accumulation was detected in Boea leaves treated with dithiothreitol (DTT) and Boea leaves in the dark, and after 3 d rehydration, the parameters Fv/Fm,ΦPSⅡ, qP and NPQ showed full recovery in control Boea leaves under very weak light condition, but the parameters only underwent partial recovery in Boea leaves treated with DTT and Boea leaves in the dark, suggesting that the recovery of photosystemⅡ (PSⅡ) photochemical activities in Boea leaves was obviously affected by treatments with DTT and darkness, therefore, zeaxanthin may play an important protective role in desiccated Boea leaves even under very weak light conditions. Key words: chlorophyll fluorescence; desiccation; resurrection angiosperm; xanthophyllcycle
阳文龙 胡志昂 王洪新 单际修 匡廷云
The embryogenesis, pollen germination, floral character and seed physiology of the endangered plant Manglietia aromatica Dandy were investigated. Based on this study, this species has very low seed set rate. The abortion rate of functional megaspores in all the ovules is 27.9%, the egg cell abortion rate of mature embryo sacs is up to 80%, and the germination rate of pollen grains is as low as nearly 0.01%. In addition, the floral structure appears to be another limited factor for the effective pollination of this species. The endangerment mechanism of this species seems to be comprehensive. Human’s destroying actions are the direct factors that have made the population degenerate quickly; low reproductive ability and the destroyed environments are the main reasons that prevent the population from renovating and spreading. Therefore, the conservation measures suggested by this study are to research the breed technology, artificial population renovating, in situ conservation, and ex situ conservation. Key words: endangered plant; Manglietia aromatica; reproductive biology; endangerment mechanisms
潘跃芝 梁汉兴 龚 洵
摘要： 针对香木莲（Manglietia aromatica Dandy)结籽率低以及野外实生苗稀少的现象。本文研究了香木莲雌配子体发育过程，花粉萌发力，开花生物学特性与种子结构的观察。结果表明，在雌配子体发育阶段存在以下退化现象：1）从大孢子母细胞开始减数分裂到功能大孢子形成阶段，在此过程中珠心内的功能结构发生退化，功能大孢子未能正常形成，仅残留有退化痕迹，退化率为27．9％；2）胚囊成熟时，受精前卵器细胞发生退化，退化率为80％，花粉萌发实验结果表明，在人工培养条件下具有正常萌发能力的花粉不足0．01％，这些都是香木莲结籽率低的重要原因，同时，香木莲的开花生物学特性限制了传粉的顺利进行。调查和研究的结果显示，人为的砍伐是造成香木莲种群急剧减少的直接原因；有性生殖障碍和生境破坏是制约香木莲种群更近的最主要因素，由此提出了对香木莲的拯救和保护对策，就地保护；研究并推广繁育技术，进行种群重建；进行迁地保护，保存尽可能多的种质资源。 (共6页)
通讯作者。E-mail: gongx @ .mail.km169.net。
Processing of pre-rRNA is one of the major events taking place in the nucleolus. U3 snoRNA, an rRNA spliceosomal factor, is suggested to be essential in the first cleavage step of the 5′ ETS sequence in the processing of pre-rRNA. Identification of U3 in the nucleolus provides a piece of indirect evidence for pre-rRNA processing site and transportation of processing products. In the present study, subnucleolar distribution of U3 snoRNA in the nucleolus of Pisum sativum L. was studied by in situ hybridization with a U3 snoRNA probe. The results showed that the U3 labeling signals were distributed throughout dense fibrillar components (DFCs) and granular components (GCs), while no signal was found in fibrillar centers (FCs). When treated with actinomycine D (AMD), the labeling signals were decreased. Along with the increase of the AMD treatment time, the labeling signals became fewer and they were found in the distal regions of DFC and GC. Our results indicated that pre-rRNA splicing took place in the regions of DFC and GC, and the transportation of pre-rRNA processing products was from the regions around FCs towards the distal regions. Key words: Pisum sativum; nucleolus; rRNA; processing; snoRNA
龙 鸿 曾宪录 胡 波 孙海晶 刘振兰 郝 水
（1. 东北师范大学遗传与细胞研究所，长春130024; 2. 南开大学生命科学学院，天津300071）
摘要： rRNA前体剪切是发生在核仁中的重要生物学事件.U3 snoRNA作为rRNA的一个剪切因子被认为是rRNA前体剪切第一步,即5′ ETS剪切所必需的.鉴定U3能够为确定rRNA前体剪切位点和剪切产物转运提供间接证据.本文利用原位杂交技术研究了豌豆(Pisum sativum L.)核仁中U3 snoRNA的分布和转运.结果表明, U3 snoRNA分布在致密纤维组分(dense fibrillar component, DFC)和颗粒组分(granular component, GC)中,在纤维中心(fibrillar center, FC)没有分布.当用放线菌素D (actinomycin D, AMD)处理豌豆根端分生细胞时,rDNA转录受到抑制,标记信号减弱.随着AMD处理时间的延长,标记信号逐渐变弱并出现在DFC远轴区域和GC区域.本文结果提示,rRNA前体剪切发生在DFC和GC区域,剪切产物从围绕FC的区域向周边转运.
通讯作者。Tel: 0431-5665130; Fax: 0431-5687517; E-mail: ycsuo @ nenu.edu.cn。
The ultrastructures of the root protophleom sieve element at different developmental stages of Arabidopsis thaliana L. were investigated using the technique of high pressure freezing and freeze substitution fixing specimen. The results show that in the development of the sieve elements, the nuclei undergo typical characteristics of the programmed cell death (PCD): the nuclear envelopes form emboli, the chromatin condenses and aggregates towards the nuclear envelope, which degrades and fully disappears later. Before the nucleus degradation， neither the nuclear envelope undulation, nucleus lobe nor marked dilation (or bleb) of perinuclear space could be observed. In the cytoplasm of the mature sieve element, there are starch-like granules separately sheathed with a layer of membrane and usually with mitochondria around. These granules seem to provide substrates to mitochondria in their function. Small vacuoles originate from endoplasmic reticulum (ER), and no bigger vacuole was found. Key words: Arabidopsis thaliana; root; sieve element; ultrastructure; high pressure freezing
Dehydration-responsive element-binding (DREB) proteins specifically binding with dehydration-responsive element (DRE) have been identified as a kind of important transcription activator of plants under drought, high salt and cold stress. The conserved amino acid residues of Val (14th residue) and Glu (19th residue) in AP2/EREBP domain of DREB1A have been identified to be two key points in determining the binding ability of DREB gene with DRE element. Using the yeast one-hybrid system, we isolated one maize DREB gene named maDREB1 by screening cDNA library.Trans-activation experiment in yeast reporter strain demonstrated that maDREB1 protein could function as a DREB transcription factor activating target gene expression by specifically binding to the DRE cis-element. To assess the functional significance of these two residues in maDREB1, the V14 and E19 were substituted individually or doubly by Ala and Asp. Point mutation analysis showed that V14 substitution made significant loss of binding ability with DRE element, while point mutation of E19 had less effect. If the substitution happened simultaneously to these two residues, it would lead to great loss of the ability of binding with DRE element. It suggested that V14 and E19 were both important in protein-DNA interacting in maDREB1, though 14V was more essential. The copy number and expression pattern of maDREB1 was discussed. Key words: maDREB1; transcription factor; dehydration-responsive element (DRE)
秦 峰 李 洁 张贵友 赵 军 陈受宜 刘 强 !
（1. 清华大学生物科学与技术系，北京100084；2. 中国农业科学研究院生物技术研究中心，北京100081；
摘要： DREB类的转录因子特异性地与DRE元件（脱水应答元件）结合，在植物感受干旱，高盐及低温等逆境条件时，激活一系列下游逆境应答基因的表达。进一步的研究发现，拟南芥DREB蛋白的DNA结合域（AP2区）中14位的缬氨酸和19位的谷氨酸对该转灵因子与DNA结合起着关键性的作用。利用酵母单杂交的方法，我们从玉米（Zea mays L.)的cDNA文库中分离到一个编码与DRE元件结合的蛋白的基因，命名为maDREB1。酵母体内的反式激活实验表明，该基因编码和蛋白能特异地与DRE元件结合并能激活下游报告基因的表达。对maDREB1蛋白14位和19位的氨基酸进行单位突变和双点突变实验，发现14位的缬氨酸突变为丙氨酸后maDREB1几乎丧了其转录激活能力，而19位的谷氨酸突变为天门冬氨酸后maDREB1的转录激活能力也受到较大影响。
通讯作者。E-mail:liuq @ mail. tsinghua.edu.cn。
Two pairs of degenerate primers were designed based on nucleotide-binding site (NBS) and serine/threonine kinase domain. PCR was performed with the primers and cDNA from the Triticum aestivum-Haynaldia villosa translocation line 6VS/6AL. Amplified products were cloned and sequenced. Nine clones with NBS and one with serine/threonine kinase domain were obtained. The NBS clones were classified to six groups according to their nucleotide sequence identities (90% or higher). These resistance gene analogs (RGAs) all have open reading frames (ORF), and their amino acid sequences show high similarity to Yr10 in wheat, Mla1 and Mla6 in barley, RPS2 in Arabidopsis and other resistance (R) genes with conserved motifs. They were preliminarily mapped on the chromosomes of homoeologous groups 1, 2 and 5 of common wheat by nulli-tetrasomic analysis. The 5′-end sequence of an RGA N5 was obtained by 5′-RACE PCR. It encodes six leucine zipper (LZ) and has high sequence similarity to RPS2. Key words: resistance gene analogs; nucleotide-binding site; PCR; Triticum aestivum-Haynaldia villosa translocation line
根据 R 基因保守区分离小麦R 基因类似序列
秦跟基 陈佩度 顾红雅 冯 高 牛吉山
摘要： 用根据核苷酸结合位点(nucleotide binding site, NBS)和丝氨酸/苏氨酸蛋白质激酶域设计的2对简并性引物,以小麦-簇毛麦6VS/6AL易位系的cDNA为模板进行PCR扩增.扩增产物克隆到pGEM-T载体中,经测序,共获得具有NBS结构域特征的片段克隆9个和具有丝氨酸/苏氨酸蛋白质激酶域特征的片段的克隆1个.将克隆之间核苷酸序列同源性高于90%的克隆归为一类,把9个NBS片段分为6类.这6类抗病基因类似序列(resistance gene analogs, RGA)均具有阅读框,并与已克隆的小麦抗条锈病基因Yr10、大麦抗白粉病基因Mla1和Mla6、拟南芥的抗病基因RPS2以及其他一些抗病基因在NBS保守区内具有高度的同源性.用小麦中国春缺体-四体初步将它们分别定位于小麦第一、第二和第五部分同源群上.进一步用5′-RACE技术获得RGA N5的5′-端,发现其编码产物的N端还具有6个亮氨酸拉链(leucine zipper, LZ),与RPS2的N-端有较高同源性.
关键词： R 基因类似片段；核苷酸结合位点；:PCR；小麦-簇毛麦6VS/6AL易位系
通讯作者。Tel: 025-4396026; Fax: 025-4395344;E-mail: pdchen @ njau.edu.cn.
A gene with complete sequence of 1 176 bp containing an open reading frame (ORF) of 281 amino acids, RSG6 (accession number in GenBank: AF442490) was cloned from rice (Oryza sativa L.) sperm cell cDNA library by using sperm cell higher expression clone BF475207 as probe. It is the first gene isolated directly from sperm cells of higher plants. The sequences of RSG6 gene and its deduced amino acid did not reveal remarkable similarity to that of any known ones in GenBank. Southern hybridization suggested that there may be introns in this gene. Reverse transcriptase-polymerase chain reaction analysis (RT-PCR) showed that the RSG6 gene was expressed in all rice organs or cell types examined (including roots, leaves, two-cell stage pollen, mature pollen, pollinated ovary and sperm cells), but the level of expression in sperm cells was particularly higher than that in all the other organs or cells. The RSG6 gene was cloned into a vector pQE30 and expressed in Escherichia coli M15 (pREP4). High specific antibody with high titer was obtained from rabbits. Key words: sperm cell; cloning; expression; rice
白彧 徐莺 唐琳 颜钫 苟小平 陈放
摘要： 用水稻 (Oryzasativa L .)精细胞优势表达克隆BF4 75 2 0 7为探针 ,筛选水稻精细胞cDNA文库 ,得到一全长为1176bp的序列 ,其开放读码框编码 2 81个氨基酸 ,与已知蛋白质无明显同源性 ,属于一新发现的基因 ,GenBank登录号为AF4 4 2 4 90。Southern杂交显示该基因可能含有内含子。RT_PCR结果显示该基因在根、叶、二细胞花粉、成熟花粉、授粉子房和精细胞中均有表达 ,但在精细胞中的表达量要高得多 ,是精细胞差异表达基因。将此基因命名为RSG6 (ricespermgene 6 )。将RSG6的编码区克隆到表达载体pQE30上 ,构建重组质粒。在大肠杆菌M15中表达出N端融合了 6×His的融合蛋白。用纯化的融合蛋白免疫家兔 ,制得高效价、高特异性的抗体。
!通讯作者。Tel: 028-85417281; E-mail: chenfang @ scu. edu.cn.
RAPDs and AFLPs were used to determine the genetic relationships among 23 elite cultivars of confectionary sunflowers (Helianthus annuus) from different districts in China. Both approaches uniquely fingerprint each of the accessions. Twenty-six RAPD primers resulted in a total of 192 strong DNA fragments, ranging from 0.26 kb to 1.98 kb, among which 165 (86.12%) were polymorphic. The average number of DNA band produced by each primer was 7.38. A total of 576 AFLP markers were produced with 8 primer combinations, ranging from 100 bp to 500 bp, and 341 polymorphic bands (59.20%) were revealed. The polymorphism rate was 76.00% and the average bands amplified by per primer combination were 72. Effective number of alleles per locus of RAPD marker (1.76) was larger than that of the AFLP marker (1.65). The mean PIC value of AFLP markers (0.38) was lower than that of the RAPD markers (0.41), but AFLP marker had much higher Ai value (38.52) than RAPD marker (6.38). Genetic similarities from RAPD data ranged from 47.84% to 82.06% and the average Nei’s coefficient was 0.649 5; the Nei’s coefficient of similarity from AFLP data ranged from 54.15% to 83.52%, and the average Nei’s coefficient was 0.6884. However, standard deviation (SD) of RAPDs was 0.13 but the SD of AFLPs was 0.08. In general, the RAPD data gave lower similarity values and higher SD values than those based on the AFLP analysis. The correlation coefficient between the two genetic similarity matrices was 0.51, revealing the estimations of genetic relationship provided by the two marker systems were only moderate. However, cluster analyses of RAPD or AFLP data divided the 23 sunflower genotypes into identical 3 groups. Key words: confectionary sunflower; RAPD; AFLP; genetic diversity
刘杰 刘公社 Chao Chien JAN
2. United States Department of Agriculture,Northern Crop Science Laboratory, Fargo, ND 58105, USA
摘要： 本研究采用RAPD和AFLP方法对23个中国不同地区的食用向日葵(Helianthus annuus L.)骨干品种进行了遗传变异分析,同时对两种标记系统进行了比较.26个RAPD引物产生了总计192条DNA条带,大小分布于0.26 kb～1.98 kb之间,其中165条(86.12%)具有多态性,每条引物产生DNA条带的平均数为7.38.8对AFLP引物组合共产生了576条带,分布于100 bp～500 bp之间,其中的341条具有多态性,多态百分率为76.00%,每对引物组合产生DNA条带的平均数为72.RAPD方法检测到的每位点有效等位基因数(1.76)大于AFLP(1.65),AFLP标记位点的平均多态性信息量(PIC)(0.38)低于RAPD标记位点的PIC(0.41),但AFLP标记具有很高的多态性检测效率(Ai=38.52).用RAPD标记分析23个食用向日葵材料的亲缘关系,Nei氏相似性系数分布在47.84%～82.06%,平均相似性系数为0.649 5, 而采用AFLP的Nei氏相似性系数分布在54.15%～83.52%,平均相似性系数为0.688 4.RAPD数据的标准差为0.13,而AFLP数据的标准差为0.08.因此,采用RAPD和AFLP方法分析食用向日葵遗传变异,RAPD标记具有较低相似性系数和较高方差而AFLP则相反.源于两种不同标记的遗传相似性矩阵的相关系数为0.51,说明采用RAPD和AFLP系统分析食用向日葵遗传变异得到的结果有一定的相关性.无论采用RAPD还是AFLP标记进行聚类分析,都将23个不同基因型的食用向日葵材料分成了三个类群.
A genomic DNA fragment containing the 5′-upstream sequence and part of the open reading frame corresponding to the cytosolic fructose-1,6-bisphosphatase (cyFBPase) cDNA was isolated by Genome Walking. The 1 195 bp 5′-flanking region which started from the translation initiation ATG codon was fused to reporter gene encoding β-glucuronidase (GUS) and stably transferred to rice via particle bombardment. Strong GUS activity was detected in leaves and leaf sheaths of transgenic rice, but not in culms and roots. Histochemical localization revealed that GUS expression was exclusively restricted to mesophyll cells in transgenic rice. Our results indicate that the 1 195 bp fragment contains all the cis-elements required for directing mesophyll-specific expression pattern in rice. Key words: rice (Oryza sativa)； promoter； cytosolic fructose-1,6-bisphosphatase gene; mesophyll-specific expression
水稻细胞质1，6-二磷酸果糖酶基因1 195 bp 的上游调控序列的克隆及其在转基因水稻中的表达研究
司丽珍 曹守云 储成才
通讯作者。E-mail: ccchu @ genetics.ac.cn。
Two new antioxidant compounds, securixanthones C (1) and D (2) were isolated from the stems of Securidaca inappendiculata Hassk. The structures of the two new compounds were established on the basis of EI-MS, 1D-NMR and 2D-NMR, UV, and IR experiments. Key words: Securidaca inappendiculata; Polygalaceae; securixanthone C; securixanthone D
杨学东 徐丽珍 杨世林
摘要：首次从蝉翼藤(Securidaca inappendiculata Hassk)茎乙醇提取物的乙酸乙酯部分分离到两个新的具有抗氧化作用的口山酮类化合物securixanthone C (1) 和 securixanthone D (2).这两个新化合物的结构是通过波谱方法(1D-NMR和2D-NMR、 EI-MS、 UV 和IR)鉴定的.
Two new tirucallane-type triterpenoid saponins, Sapimukoside A (1) and Sapimukoside B (2), were isolated from the roots of Sapindus mukorossi Gaertn. On the basis of spectroscopic and chemical evidence, their structures were elucidated to be 3-O-α-L-rhamnopyranosyl-(1→2)-［α-L-arabinopyranosyl-(1→3)］-β-D-glucopyranosyl-21, 23R-epoxyl tirucall-7, 24R-diene-3β, 21-diol（1） and 3-O-α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranosyl-21, 23R-epoxyl tirucall-7,24R-diene-3β, 21-diol（2）. Key words: Sapindus mukorossi; Sapindaceae; triterpenoid saponin; Sapimukoside A; Sapimukoside B
滕荣伟 倪伟华 燕陈昌祥
从无患子科无患子（Sapindus mukorossi Gaertn.)根中分离到两个新的大戟烷型三萜皂甙SapimukosideA（1）和SapimukosideB（2）。运用波谱和化学方法鉴定它们的结构为3－O－α-L鼠李吡喃糖基－（1→2）-[α-L－阿拉伯吡喃糖基－（1→3）]－β－D－葡萄吡喃糖基－21，23R－epoxyl tirucall-7,24R-diene-3β，21-diol(1)和3－O－α-L－鼠李吡喃糖基－（1→6）－β-D-葡萄吡喃糖基21，23R-epoxyl tirucall-7,24R-diene-3β，21-diol(2)。
关键词：无患子 无患子科 三萜皂甙：Sapimukoside A； SapimukosideB
通讯作者。) Tel：0871-5223243；Fax：0871-5219934；E-mail：< cxchen @ mail.kib.ac.cn> 。
A high taxol yield cell line of Taxus yunnanensis Cheng et L. K. Fu keeps a high taxol-producing level after successive subcultures for more than eight years. In this study, eight taxanes were isolated from the suspension cell cultures of this cell line. Based on NMR and MS analyses, and comparison with literature data and standards, their structures were determined to be 2α,5α,10β-triacetoxy-14β-propionyloxy-4(20),11-taxadiene (1), 2α,5α,10β-triacetoxy-14β-(2′-methyl)-butyryloxy-4(20),11-taxadiene (2), 2α,5α,10β-14β-tetra-acetoxy-4 (20),11-taxadiene (3, taxuyunnanine C), 2α,5α,10β-triacetoxy-14β-(2′-methyl-3′-hydroxy)-butyryloxy-4(20),11-taxadiene (4, yunnanxane) and its 3′-epimer (5), baccatin Ⅳ (6), baccatin Ⅲ (7) and taxol (8),respectively. Among those compounds, 3, 5, 6 and 7 were reported to be isolated from the suspension cell cultures of T. yunnanensis for the first time. TLC and HPLC analyses indicated that the chemical constituents of the culture solution were similar to those of cultured cells. Moreover, the highest taxol content of this cell line reached 0.3% and the cell line could be applied for a large-scale culture. Key words:Taxus yunnanensis; cell suspension cultures; taxol; taxanes
摘要：云南红豆杉(Taxus yunnanensis Cheng et L. K. Fu)的一株紫杉醇高产细胞系经过8年多的继代培养,仍保持较稳定的紫杉烷类化合物的生物合成能力.从此株紫杉醇高产细胞系的悬浮培养物中分离到8个紫杉烷类化合物,经核磁共振光谱和质谱数据分析,它们的化学结构分别是2,5,10-三乙酰氧基-14-丙酰氧基紫杉二烯(1)、 2,5,10-三乙酰氧基-14-(2′-甲基丙酰氧基)紫杉二烯(2)、 2,5,10,14-四乙酰氧基紫杉二烯(3)、 2,5,10-三乙酰氧基-14-(2′-甲基-3′-羟基丁酰氧基)紫杉二烯及其差向异构体(4和5)、巴卡亭Ⅳ(6)、巴卡亭Ⅲ (7)和紫杉醇(8).化合物3、 5-7为首次从云南红豆杉细胞培养物中分离到.定性分析表明,云南红豆杉细胞悬浮培养液中的化学成分与培养细胞中的相似.另外,此株紫杉醇高产细胞系的紫杉醇含量可高达0.3%,可用来进行大规模培养.
通讯作者。Tel:0871-5223111;Fax:0871-5150227; E-mail:yshen @ public. km.yn.cn
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