Inheritance and Resistance to Insect in CryIA(c) Transgenic Cabbage

以结球甘蓝（Brassica oleracea var. capitata ）下胚轴为外植体，通过根癌农杆菌（Agrobacterium tumefaciecin）介导法将苏云金芽孢杆菌（Bacillus thuringiensis,Bt）杀虫蛋白基因CryIA(c)导入结球甘蓝栽培品种中，共获得56株卡那霉素抗性植株。分别以CryIA(c)和NPTⅡ基因引物进行PCR扩增，结果43株均扩增出了预期的目的带。以CryIA(c)基因为探针，进行Southern blot分析，证明外源基因已经整合到甘蓝基因组中，且多数转基因植株只含有单拷贝的外源基因。离体叶片抗虫实验表明，转基因植株幼虫校正死亡率为40%，平均单虫重达到极显著水平，叶片损害程度差异明显，转基因植株抗性较对照显著提高。通过卡那霉素涂抹叶片和PCR扩增两种方法鉴定，表明外源基因在转基因甘蓝自交后代呈孟德尔单基因遗传。     

转柽柳金属硫蛋白基因（MT1）烟草的获得及对重金属镉的抗性分析

将柽柳（Tamarix. sp）金属硫蛋白基因（MT1）插入到植物表达载体pBI121，利用农杆菌（Agrobactrium tumefaciens）介导法将MT1导入烟草基因组。对转基因T1植株的卡那霉素抗性分析表明，多数转基因植株后代表现为3:1的分离比例，只有少数转基因植株的抗性分离表现为15:1或不符合孟德尔遗传的分离比例。说明整合到烟草基因组的外源基因多为单拷贝基因，也有少数为多拷贝基因。对具有卡那霉素抗性的转基因植株进行PCR-Southern检测和Northern杂交分析表明，外源MT1基因已整合到烟草基因组，并且得到了正确表达。转基因植株对重金属镉的抗性比对照显著提高，表现为转基因植株的株高和鲜重均明显优于非转基因株系。     

根癌农杆菌介导的甘蓝高效稳定的遗传转化系统的建立及对CpTI基因转化的研究

通过对影响根癌农杆菌转化的多种因素的比较和探索后，建立了一种以农杆菌介导的高效和稳定的甘蓝遗传转化系统。选择目前在生产上大量推广应用的6个优良甘蓝品种，用其无菌苗的下胚轴作为外植体，经《农杆菌LBA4404（含质粒pBin-TI-19-2)感染，在含卡那霉素50mg/L的抗性培养基上筛选，80％的外植体表现出抗性，形成愈伤组织并进一步分化成苗。每块愈伤组织最低成苗5株，最高可达22株。对部分抗性植株的总DNA进行Southern杂交，结果表明T－DNA上的CpTI基因已整合到甘蓝基因组中，外源基因的转化频率高达20％。通过抗小菜蛾实验发现，转基因植株未转基因植株有明显的抗性。     

商陆抗病毒蛋白基因转化芥菜的初步研究

以芥菜(Brassica Juncea Coss.)的子叶为外植体,通过根癌农杆菌（Agrobacterium tumefaciens）的介导,将商陆抗病毒蛋白（PAP）基因导入到芥菜中。对所获得的40株抗性植株进行PCR扩增,其中阳性植株为29株;Southern blot分析结果证明PAP基因已被整合到芥菜基因组中,在大多数转基因植株中外源基因呈单挎贝,少数为双挎贝;Northern blot分析结果表明,PAP在转基因植株中正常表达。转基因植株接种病毒试验初步结果表明,转PAP基因的植株对TuMV的侵染有一定的抑制作用。     

转透明颤菌血红蛋白基因杨树的再生及其生长量初步测定

采用农杆菌介导法,将透明颤菌血红蛋白Vitreoscilla hemoglobin(VHb)基因vgb导入银腺杨(Populus alba ×P.glandulosa).以经无菌培养的叶片为外植体,通过450个叶盘与根癌农杆菌(Agrobacterium tumefaciens)LBA4404共培养,将植物双元表达载体pPZV中vgb基因导入银腺杨基因组,经卡那霉素筛选后,共获得60株卡那霉素抗性植株.经PCR特异性扩增和Southern点杂交分析,证明其中52株再生植株基因组DNA中整合了vgb基因.RT-PCR分析结果表明,vgb基因在其中的44个株系中获得表达.在2年温室生长条件下,转基因无性系的外部形态与对照相比无显著差异.但在株高和地径上,有3株转基因无性系较对照植株有明显增加.     

RPW2表达载体构建及对烟草的遗传转化

将RPW2基因插入表达载体pBI121的表达盒中,构建了RPW2组成型表达载体。通过农杆菌叶盘转化法转化烟草叶片,经卡那霉素筛选、PCR扩增和Southern杂交鉴定证明,RPW2基因已成功转入烟草细胞中,获得6个转基因株系。RT-PCR半定量分析表明,转基因烟草株系的叶片中有RPW2基因的表达。不同转基因株系表达量不同,转基因株系T-R2和T-R4表达量较高。转基因植株叶片上人工接种烟草赤星病菌液检测转基因植株的抗病性,结果表明,与对照植株相比转RPW2基因植株对烟草赤星病抗性显著增强,说明RPW2基因在烟草中也具有抗病能力。另外,转基因株系间RPW2基因表达量虽有差异,但抗病性没有显著差异。     

双Bt基因提高青花菜对菜粉蝶和小菜蛾抗性

以青花菜（Brassica oleracea L.ssp.italica）下胚轴为外植体,通过农杆菌介导的遗传转化方法将苏云金芽胞杆菌（Bacillus thuringiensis）晶体毒蛋白基因Cry1Ac和Cry1C分别导入青花菜栽培品种绿彗星中,各获得35株卡那霉素抗性植株。分别以Cry1Ac和Cry1C特异引物对转基因植株进行PCR扩增,结果得到27株Cry1Ac阳性植株和13株Cry1C阳性植株。以目的基因Cry1Ac和Cry1C为探针,进行Southernblot分析,证明多数植株目的基因已经整合到青花菜基因组中。对含有单拷贝目的基因植株自交纯合后通过有性杂交获得带有Cry1Ac和Cry1C双价的转基因青花菜。利用RT-PCR检测Bt基因在转基因植株中的表达情况,发现不同株系间在RNA水平上存在显著的表达差异,部分株系中双价Bt同时得到表达。ELISA检测转基因植株Bt毒蛋白含量为0.12～0.82μg/gFW。结合室内和田间饲虫试验,证明抗虫效果与Bt毒蛋白表达水平正相关。通过多世代选择,获得高抗菜粉蝶（Pieris rapae L.）和小菜蛾（Plutella xylostella L.）的纯合转基因青花菜材料。双价抗虫基因共同作用,提高了青花菜的抗虫性。     

转5-烯醇式丙酮酰莽草酸-3-磷酸合酶(EPSPS)基因抗草甘膦烟草和棉花的获得

以从抗草甘膦的荧光假单胞菌(Pseudomonas fluorescens)G2中克隆的、并按双子叶植物偏爱密码子改造的5-烯醇式丙酮酰莽草酸-3-磷酸合酶(EPSPS)基因aroAG2M为目的基因,用菊花Rubisco小亚基的启动子驱动,在基因的5'端加叶绿体定位信号肽,构建植物表达载体。用根癌农杆菌(Agrobacterium tumefaciens)介导的叶盘法转化烟草(Nicotiana tabacum)获得转基因植株,PCR检测表明,外源基因已整合到烟草基因组中。转基因和非转基因植株6～8叶龄苗的叶片涂抹不同浓度的草甘膦异丙胺盐,表明转基因植株可抗4‰浓度的草甘膦,而非转基因对照植株则在2‰草甘膦时即死亡。花粉管通道法转化棉花(Gossypium hirsutum),得到3株具有草甘膦抗性的转基因植株,PCR和Southern检测显示,外源基因已整合到棉花基因组中,田间喷洒草甘膦异丙胺盐水剂,表明T1代转基因植株具有草甘膦抗性。     

冷诱导基因CBF-4转化辣椒与转基因植株抗寒性分析

以4个辣椒（Capsicum annuum L. ）品种为试材,利用根癌农杆菌介导的带柄子叶转化法将拟南芥CBF-4基因转入辣椒,通过调整预培养、共培养以及延迟培养时间等因素,优化并完善了甜椒品种B12的遗传转化体系,并对获得的60株卡那霉素抗性再生植株进行PCR和RT-PCR检测表明,46株为阳性转基因植株,转化率高达76.67%。经过低温处理后,对阳性植株的抗寒性生理、形态指标分析发现,转基因植株的SOD、POD和MDA活性都明显高于未转基因植株,转基因植株比未转基因植株的株型紧凑,叶片肥厚、平直,叶色浓绿,叶面积大。     

农杆菌介导法获得大量转双价抗虫基因水稻植株

构建含Bt杀虫基因cryIA(c)和豇豆胰蛋白酶抑制剂基因CpTI的双价抗虫转基因载体PCAM-Bt-CpTI，并用于农杆菌（Agrobacterium tumefaciens)介导法对粳稻品系“浙大19”遗传转化，约2000块盾片愈伤组织与农杆菌共培养后，得到了约1300块潮霉素抗性愈伤，从中分化抗性再生苗约1500株，对不同抗性愈伤来源的70个T0代再生株进行PCR及PCR-Southern检测，62株为转cryLA(c)和CpTI双价抗虫基因植株，阳性植株比例为88.6%，抗虫鉴定表明，3个转基因T1代株系对二化螟有很高的毒性。     

低氮和干旱胁迫对富士和秦冠生长及氮素利用的影响

【目的】以富士（Fuji）、 秦冠（Qinguan）嫁接在平邑甜茶（Malus hupehensis Rehd.）上的当年生盆栽苗为试验材料,采用砂培方法,研究了缺氮胁迫和干旱对富士和秦冠生长情况、 光合参数、 植株各部位氮磷钾含量及氮素利用效率的影响,分析比较了低氮干旱条件下富士和秦冠生长及氮素利用的差异,以期为果树生产高效肥水利用提供理论指导。【方法】试验共设四个处理: 正常氮正常水（ZZ）、 低氮正常水（DZ）、 正常氮干旱（ZG）、 低氮干旱（DG）。氮素和水分均设置两个水平,分别为正常氮（6 mmol/L NO-3-N）、 低氮（0.3 mmol/LNO-3-N）、 正常供水（保持盆中砂子相对含水量为饱和含水量的80%~85%）、 干旱处理（保持盆中砂子相对含水量为饱和含水量的60%~65%）。【结果】富士和秦冠的生物量（茎和叶）、 株高茎粗等生长指标以及光合速率、 气孔导度、 蒸腾速率均为正常氮正常水（ZZ）＞低氮正常水（DZ）＞正常氮干旱（ZG）＞低氮干旱（DG）,并且相对应处理下秦冠的以上指标均高于富士;正常供水下,缺氮处理使富士、 秦冠的根冠比比正常氮处理均有所增加,富士提高了2.05%,秦冠提高了22.40%。富士和秦冠的氮、 磷、 钾含量均表现出正常氮正常水（ZZ）＞低氮正常水（DZ）＞正常氮干旱（ZG）＞低氮干旱（DG）; 氮、 钾元素含量在植株各部位的分布顺序依次是叶＞根＞茎,磷元素则是根＞叶＞茎;光合氮素利用效率（PNUE）和氮素利用效率表现为秦冠处理之间差异极显著,富士处理之间差异不显著;秦冠的PNUE和NUE明显高于富士,在低氮正常水（DZ）处理下,秦冠氮肥利用率比富士高42.07%,在低氮干旱（DG）处理下高64.14%;低氮胁迫下富士和秦冠的NUE显著提高,并且秦冠提高的幅度高于富士。【结论】施用氮肥能够显著提高富士与秦冠的干物质量,同等水肥条件下,秦冠生长优于富士;水分亏缺会减少叶片对氮的吸收,干旱条件下适度增施氮肥,可提高果树的抗旱能力;低氮干旱胁迫下秦冠的生长指标、 光合指标及氮素利用效率指标均优于富士,表现出较强的抗低氮干旱胁迫的能力。     

Pasture Vegetation Elemental Analysis by Laser-Induced Breakdown Spectroscopy

Laser-induced breakdown spectroscopy (LIBS) is a new technique for the analysis of plant material. This study investigates the application of LIBS to pasture-based plant samples. The LIBS measurements were obtained from pelletized pasture samples (100 samples) that had been also analyzed by inductively coupled plasma–optical emission spectroscopy (ICP-OES) following microwave digestion for calibration and comparison purposes. Comparisons for elements sodium (Na), potassium (K), magnesium (Mg), calcium (Ca), manganese (Mn), iron (Fe), copper (Cu), zinc (Zn), boron (B), phosphorus (P), and sulfur (S) showed that LIBS could be used for almost all the standard profile total elements with concentrations down to low mg/kg levels (observed error of Na: 0.024 percent, K: 0.18 percent, Mg: 0.016 percent, Ca: 0.073 percent, P: 0.017 percent, Mn: 31 mg/kg, Fe: 150 mg/kg, Zn: 6.6 mg/kg, and B: 1.1 mg/kg). Elemental analysis at less than mg/kg levels was not possible using LIBS. The elements S and Cu were particularly difficult to analyze with reliability using LIBS at the concentration levels found in the plant samples. Replacing microwave digestion and subsequent ICP analysis with a direct analysis of dried plant samples using LIBS has the potential to improve the productivity and reduce the cost of testing.     

Faculty positions:Center for Agricultural Resources Research,Chinese Academy of Sciences

正The Center for Agricultural Resources Research(CARR),the Institute of Genetics and Developmental Biology(IGDB),Chinese Academy of Sciences,invites applicants for several research group leader positions.CARR is one of the research organizations in Chinese Academy of Sciences(CAS).We seek nominations and applications from individuals who have expertise and a record of accomplishment in research areas related to ecology,agro-hydrology,     

Macroinvertebrates in North American tallgrass prairie soils: effects of fire, mowing, and fertilization on density and biomass

The responses of tallgrass prairie plant communities and ecosystem processes to fire and grazing are well characterized. However, responses of invertebrate consumer groups, and particularly soil-dwelling organisms, to these disturbances are not well known. At Konza Prairie Biological Station, we sampled soil macroinvertebrates in 1994 and 1999 as part of a long-term experiment designed to examine the effects and interactions of annual fire, mowing, and fertilization (N and P) on prairie soil communities and processes. For nearly all taxa, in both years, responses were characterized by significant treatment interactions, but some general patterns were evident. Introduced European earthworms (Aporrectodea spp. and Octolasion spp.) were most abundant in plots where fire was excluded, and the proportion of the total earthworm community consisting of introduced earthworms was greater in unburned, unmowed, and fertilized plots. Nymphs of two Cicada genera were collected (Cicadetta spp. and Tibicen spp.). Cicadetta nymphs were more abundant in burned plots, but mowing reduced their abundance. Tibicen nymphs were collected almost exclusively from unburned plots. Treatment effects on herbivorous beetle larvae (Scarabaeidae, Elateridae, and Curculionidae) were variable, but nutrient additions (N or P) usually resulted in greater densities, whereas mowing usually resulted in lower densities. Our results suggest that departures from historical disturbance regimes (i.e. frequent fire and grazing) may render soils more susceptible to increased numbers of European earthworms, and that interactions between fire, aboveground biomass removal, and vegetation responses affect the structure and composition of invertebrate communities in tallgrass prairie soils.     

Variation in quality parameters between and within 14 Nordic tree fruit and berry species

Abstract

A 3-year study was carried out to investigate quality parameters in 14 tree fruit and berry species grown in southern Norway. The species were blueberry, apple, aronia, sour cherry, sweet cherry, red raspberry, strawberry, blackcurrant, gooseberry, red currant and elderberry, harvested along with wild bilberry, cloudberry and lingonberry. Significant differences between species were identified for all quality parameters. The coefficient of variation between species was lowest for pH (12.5%), dry matter (18.9%) and soluble solids (25.3%), followed by titratable acids (59.3%), total phenolics (83.8%), antioxidant capacity FRAP (85.7%) and antiradical power by the DPPH-assay (97.8%), total monomeric anthocyanins (132%) and ascorbic acid (137%). Average coefficient of variation within species were lower and ranged from 4 (pH) to 62% (ascorbic acid). Only the FRAP values were significantly affected by harvesting year with lower levels in 2004 than in 2005 and 2006. There were significant interactions between species and harvesting year for dry matter, soluble solids, pH, ascorbic acid and FRAP. The results indicate generic ranges in composition within species independent upon growing location and climate, and the composition of the tree fruits and berries is not likely to deviate from these ranges. It is concluded that desirable composition of tree fruits and berries and their products should primarily be achieved by selection among species rather than searching fors broadened variation within individual species.     

Potassium Fixation and Release Characteristics in Rhizosphere and Nonrhizosphere Soils for a Rapeseed–Rice Cropping Sequence

Potassium (K) fixation and release in soil are important factors in the long-term sustainability of a cropping system. Changes in K concentration and characteristics of K fixation and release in rhizosphere and nonrhizosphere soils in the rapeseed (Brassica napus L.)–rice (Oryza sativa L.) rotation were investigated using a rhizobox system. The concentrations of different forms of K in both rhizosphere and nonrhizosphere soils decreased with plants compared to without plants, regardless of K fertilizer application. Potassium uptake by crops mainly came from the rhizosphere soil. In the treatment without K fertilizer (–K), the main form of K supplied by the soil to the crops was 1.0 mol L^?1 nitric acid (HNO₃) nonextractable K, followed by nonexchangeable K, and then exchangeable K. In the treatment with K fertilizer (+K), the main K forms supplied by the soil to the crops were exchangeable K and nonexchangeable K. The amount and rate of K fixation after one cycle of the rapeseed–rice rotation was greater in rhizosphere soil than in nonrhizosphere soil. The amount and rate of K fixation of soil in the +K treatment were significantly less than in the –K treatment. The cumulative amounts of K released with 1.0 mol L^?1 ammonium acetate (NH₄OAc) and 1.0 mol L^?1 HNO₃ extraction increased with the increasing numbers of extractions, but the K-releasing power of soil by successive extraction decreased gradually and finally became almost constant. The release of K was less in rhizosphere soil than in nonrhizosphere soil. The release of K in the +K treatment was similar to that in the –K treatment in rhizosphere soil, but the K release in nonrhizosphere soil was greater with the +K than the –K treatment. Overall, the information obtained in this study will be helpful in formulating more precise K fertilizer recommendations for certain soils.     

Electron donor and pH relationships for biologically enhanced dissolution of chlorinated solvent DNAPL in groundwater

Biologically enhanced dissolution offers a method to speed removal of chlorinated solvent dense non-aqueous-phase liquid (DNAPL) sources such as tetrachloroethene (PCE) and trichoroethene (TCE) from aquifers. Bioremediation is accomplished by adding an electron donor to the source zone where fermentation to intermediates leading to acetic acid and hydrogen results. The hydrogen and possibly acetic acid are used by dehalogenating bacteria to convert PCE and TCE to ethene and hydrochloric acid. Reductive dehalogenation is thus an acid forming process, and sufficient alkalinity must be present to maintain a near neutral pH. The bicarbonate alkalinity required to maintain pH above 6.5 is a function of the electron donor: 800 mg/L of bicarbonate alkalinity is sufficient to achieve about 1.2 mM TCE dechlorination with glucose, 1.7 mM with lactate, and a much higher 3.3 mM with formate. Laboratory studies indicate that in mixed culture, formate can be used as an electron donor for complete conversion to ethene, contrary to pure cultures studies indicating it cannot. Various strategies can be used to add electron donor to an aquifer for DNAPL dehalogenation while minimizing pH problems and excessive electron donor usage, including use of injection-extraction wells, dual recirculation wells, and nested injection-extraction wells.     

Biodiversity and ecogeography of wild Lactuca spp. in some European countries

Staff members of the Department of Botany of Palacký University in Olomouc and Gene Bank Department – Workplace Olomouc, Research Institute of Crop Production in Prague, Czech Republic, conducted an expedition in seven European countries (Austria, Czech Republic, France, Germany, Italy, the Netherlands, Switzerland) in August/September 1999 to collect wild Lactuca spp. germplasm and study its geographic distribution, ecology and biodiversity. During the mission, more than 600 locations were visited resulting in the collection of 602 seed samples (accessions) of wild Lactuca species and 13 seed samples of related genera (Chondrilla and Mycelis). Lactuca serriola f. serriola, L. serriola f. integrifolia, L. saligna and L. viminea subsp. chondrilliflora were prevalent in southern Europe (Italy, France), however, only L. serriola was common in central and western Europe (Austria, Czech Republic, Germany, Netherlands, Switzerland). The greatest diversity of Lactuca species was found in France, where also the most seed samples (165) were collected. The most characteristic habitats with a high density of Lactuca spp. populations were observed along roads and highways, grassy ditches, ruderal communities, and dust-heaps. Natural infections by powdery mildew (Erysiphe cichoracearum) and downy mildew (Bremia lactucae) on some wild Lactuca spp. were observed. Recent observations concerning the geographic distribution, population structure, habitats, and natural occurrence of diseases of Lactuca spp. are discussed. This assemblage of genetic resources of Lactuca spp. can serve as the basis of future studies of species diversification, spatial population structure, plant microevolution, domestication processes, and genetic variability of host-parasite interactions.     

Increase in auxiliary photoprotective photosynthetic pigments in wheat seedlings induced by Azospirillum brasilense

Inoculation of wheat seedlings with the plant growth-promoting bacterium Azospirillum brasilense Cd was immobilized in alginate microbeads and, without applying any stress, significantly increased the quantity of several photosynthetic pigments, such as chlorophyll a, chlorophyll b, and the auxiliary photoprotective pigments violaxanthin, zeaxanthin, antheroxanthin, lutein, neoxanthin, and β-carotene. This resulted in greener plants with no apparent visible stress. After monitoring the quantity of photosynthetic pigments for 4 weeks, we observed that inoculated plants had higher quantities of pigments in shoot and stem. The greatest difference in the quantity of all pigments between inoculated and noninoculated plants occurred in the first week of growth. Regardless of treatment, the quantity of pigments in stems was three to four times less than the quantity of these pigments in shoots. Application of Azospirillum, either as liquid inoculant or as alginate microbeads, did not alter the positive effect of the bacteria on pigment production or the positive response of the plants towards A. brasilense Cd inoculation.     

In-situ enrichment and analysis of atrazine-degrading microbial communities using atrazine-containing porous beads

We examined the community composition of microbes that colonized atrazine-containing beads buried in agricultural soils that differed in atrazine treatment history. Bacterial abundance was 5-40-fold greater in atrazine-fortified beads. In beads containing 20 mg atrazine kg⁻¹ buried in soil with a history of atrazine application (conditioned soil), the abundance of Actinobacteria increased approximately 80-fold whereas in control soil, Actinobacteria were enriched only 10-fold and the gamma-Proteobacteria and Planctomycetes increased by 60- and 25-fold, respectively. The gamma-Proteobacteria were enriched by 120- and 230-fold in beads containing 200 mg atrazine kg⁻¹ in conditioned and control soil, respectively. The results demonstrate that BioSep^® beads are a suitable matrix for recruiting a diverse subset of the bacterial community involved in atrazine degradation.