Armillaria ostoyae in managed coniferous forests in Kastamonu in Turkey

Although several Armillaria species have been reported in Turkey, there is little information about their ecology in Turkish forests. In this study, we investigated five forest stands, approximately 5–74 ha in size, in Kastamonu province in the Black Sea Region of Turkey for the presence of Armillaria species in stumps and logs. The stands were mixed Abies nordmanniana ssp. bornmülleriana and Pinus sylvestris forests managed using a selective cuttings system; the proportion of fir in the total number of stems and stumps ranged from 36 to 98%. Based on sequence analysis of the internal transcribed spacer and intergenic spacer regions of the rDNA, all rhizomorphs sampled from the stumps and logs were of Armillaria ostoyae. The size of the genets was estimated with random amplified microsatellites analysis of the isolates and ranged from single stumps to approximately 450 m². One to seven genets were found in each stand. These results indicate that the genets had arisen from spores and vegetative spread was limited on most sites.     

The Proper Supply of S Increases Amino Acid Synthesis and Antioxidant Enzyme Activity in Tanzania Guinea Grass Used for Cd Phytoextraction

Increased Cd concentrations in the environment impair plant growth, but plants properly supplied with S may develop greater tolerance to the damage caused by Cd and be used in the remediation of contaminated environments. The aim of this study was to evaluate the Cd-phytoextraction potential of Panicum maximum cv. Tanzania grown with S rates and to identify alterations in the concentrations of nutrients and amino acids and in the activity of some antioxidant enzymes under Cd stress conditions. Combinations of five S rates (0.1, 1.0, 1.9, 2.8, and 3.7 mmol L^?1) and five Cd rates (0.0, 0.5, 1.0, 1.5, and 2.0 mmol L^?1) in a nutrient solution were provided in two plant growth periods. Concentrations of N, P, and Zn increased, while K, Fe, and Mn decreased with exposure to Cd. The concentration of Ca decreased as the S supply was increased. Isoleucine, leucine, proline, and valine concentrations increased with exposure to Cd and with higher levels of S. The APX activity was higher at the highest Cd exposure level. Activity and number of SOD and GR isoforms in the roots and of CAT in the shoots of the regrown plant decreased at the highest level of contamination by Cd, which was lessened by the supply of greater S rates. Tanzania guinea grass grown with an adequate supply of S has the potential for phytoextraction of Cd-contaminated environments.     

我国畜禽饲料资源中常量元素钙含量分布的调查

目的 调查我国畜禽饲料资源中钙含量的分布,为合理利用饲料资源及精准配制畜禽饲粮提供科学依据。方法 运用统一的数据采集规范,采集了全国31个省（市、区）的37种3 862个饲料原料样品,经微波消解后,使用IRIS IntrepidⅡ等离体子发射光谱仪对其钙含量进行了测定。结果 谷物籽实中平均钙含量为457 mg·kg ^-1（范围74.5—832 mg·kg ^-1）,其中大麦的钙含量最高,玉米最低,变异系数范围为21.2%—85.3%;谷物籽实加工副产品中平均钙含量为1 090 mg·kg ^-1（范围为93.4—3 264 mg·kg ^-1）,其中小麦DDGS的钙含量最高,碎米最低,变异系数范围为19.5%—142%;植物性蛋白饲料中平均钙含量为3 987 mg·kg ^-1（范围为1 742—7 909 mg·kg ^-1）,其中菜籽粕的钙含量最高,花生粕最低,变异系数范围为0.92%—34.6%;动物性蛋白饲料中平均钙含量为14 448 mg·kg ^-1（范围为115—50 007 mg·kg ^-1）,其中鱼粉的钙含量最高,血球蛋白粉最低,变异系数范围为25.0%—239%;秸秆类饲料中平均钙含量为5 969 mg·kg ^-1（范围为1 931—13 524 mg·kg ^-1）,其中甘薯藤的钙含量最高,小麦秸最低,变异系数范围为27.6%—39.0%;牧草类饲料中平均钙含量为6 667 mg·kg ^-1（范围为4 157—13 963 mg·kg ^-1）,其中苜蓿的钙含量最高,羊草最低,变异系数范围为30.3%—94.6%;矿物质饲料中平均钙含量为30%（19.2%—41.2%）,其中贝壳粉的钙含量最高,骨粉最低,变异系数范围为2.67%—24.0%。各类饲料原料中钙含量分布规律为：矿物质饲料＞动物性蛋白饲料＞牧草类饲料＞秸秆类饲料＞植物性蛋白饲料＞谷物籽实加工副产品＞谷物籽实。同一类别不同饲料原料间钙含量差异显著（P＜0.05）。以省（区）为单位,对不同地区玉米、小麦和豆粕中钙含量进行比较,表明不同地区玉米和豆粕中钙含量存在显著的差异（P＜0.05）,而小麦中钙含量受地区性的影响较小（P＞0.05）。各省（区）玉米的平均钙含量范围为48.1—155 mg·kg ^-1,其中山西省玉米钙含量最高,辽宁省最低,变异系数范围为20.1%—321%;各省（区）小麦的平均钙含量范围为362—590 mg·kg ^-1,其中山西省小麦钙含量最高,湖北省最低,变异系数范围为8.29%—66.1%;各省（区）豆粕的平均钙含量范围为3 001—4 153 mg·kg ^-1,其中山西省豆粕钙含量最高,江苏最低,变异系数范围为1.30%—21.4%。结论 我国饲料原料中钙含量受种类及地区性影响较大。本研究结果对于生产者了解饲料原料中实际钙含量,精准配制饲粮,以确保动物的健康及高效生产具有重要意义。     

我国畜禽饲料资源中微量元素硒含量分布的调查

目的 研究我国不同地区间各种饲料原料中硒含量分布情况,以及我国畜禽基础饲粮中硒水平,从而为饲粮中合理添补硒提供依据。方法 对采自全国31个省、直辖市和自治区的7大类（谷物籽实、谷物籽实加工副产品、植物性蛋白饲料、动物性蛋白饲料、牧草类、秸秆类和矿物质饲料）37种饲料原料共3 785个饲料样品,经预处理后用MARS6高通量密闭微波消解系统进行微波消解,然后用离子色谱-电感耦合等离子体-质谱联用仪(IC-ICP-MS)进行测定。用国家标准物质猪肝粉作为参照标准,以保证测定结果的可靠性。结果 饲料原料中硒含量测定结果表明：谷物籽实（包括玉米、小麦、稻谷和大麦）平均硒含量为0.037 mg·kg ^-1（范围为0.025—0.044 mg·kg ^-1）;谷物籽实加工副产品（包括玉米蛋白粉、玉米DDGS、玉米胚芽粕、次粉、小麦麸、小麦DDGS、碎米和米糠）平均硒含量为0.071 mg·kg ^-1（范围为0.034—0.124 mg·kg ^-1）;植物性蛋白饲料（包括膨化大豆、大豆粕、菜籽粕、棉籽粕、花生粕、亚麻粕和葵花粕）平均硒含量为0.209 mg·kg ^-1（范围为0.097—0.502 mg·kg ^-1）;动物性蛋白饲料（包括鱼粉、肉粉、水解羽毛粉、肠膜蛋白粉、血浆蛋白粉和血球蛋白粉）平均硒含量为1.217 mg·kg ^-1（范围为0.611—2.220 mg·kg ^-1）;牧草类（包括羊草、黑麦草、苜蓿和青贮玉米）平均硒含量为0.062 mg·kg ^-1（范围为0.057—0.070 mg·kg ^-1）;秸秆类（包括玉米秸秆、小麦秸秆、稻秸和甘薯藤）平均硒含量为0.069 mg·kg ^-1（范围为0.033—0.128 mg·kg ^-1）;矿物质饲料（包括石粉、磷酸氢钙、骨粉和贝壳粉）平均硒含量为0.352 mg·kg ^-1（范围为0.085—0.544 mg·kg ^-1）。这37种饲料原料的平均硒含量范围为0.025—2.220 mg·kg ^-1,而各类饲料原料硒含量分布规律是：动物性蛋白饲料（1.217 mg·kg ^-1）＞矿物质饲料（0.352 mg·kg ^-1）＞植物性蛋白饲料（0.209 mg·kg ^-1）＞谷物籽实加工副产品（0.071 mg·kg ^-1）＞秸秆类（0.069 mg·kg ^-1）＞牧草类（0.062 mg·kg ^-1）＞谷物籽实（0.037 mg·kg ^-1）。以省（区）为单位比较,发现不同地区间的玉米、小麦和大豆粕的硒含量差异显著（P＜0.05）;所测省（区）玉米样品全部缺硒（≤0.05 mg·kg ^-1）,其中有61.1%的省（区）严重缺硒（≤0.02 mg·kg ^-1）;四川省小麦严重缺硒（≤0.02 mg·kg ^-1）;四川省和内蒙古自治区豆粕缺硒（0.03—0.05 mg·kg ^-1）,河南省豆粕临界缺硒（0.06—0.09 mg·kg ^-1）。根据全国各地猪、鸡常用的153个饲料配方计算出基础饲粮中可提供的硒含量为0.06—0.11 mg·kg ^-1,如根据我国猪、鸡饲养标准中硒含量的要求,基础饲粮中的硒含量仅能提供猪、鸡硒营养需要的约1/4。结论 我国不同种类和不同地区饲料原料中硒含量差异较大,全国猪、鸡常用基础饲粮配方中硒含量可提供硒营养需要量的约1/4。因此,在实际生产中,建议参考不同地区饲料原料中硒含量分布调查数据,精准配制饲粮,以满足畜禽高效生产的需要及减少硒的添加量。     

葡萄霜霉病菌实时荧光定量PCR检测体系的建立和应用

【目的】葡萄霜霉病是葡萄生产上重要的单年流行病害,研究旨在构建葡萄霜霉病菌（Plasmopara viticola）的实时荧光定量PCR（real-time PCR）检测体系,为葡萄霜霉病的早期诊断和预测预报提供依据。【方法】依据GenBank中葡萄霜霉病菌cox2基因序列设计1对特异性引物（F-cox-Pv/R-Pv）,建立并优化常规PCR和real-time PCR反应体系,利用葡萄霜霉病菌、葡萄炭疽病菌（Colletotrichum gloeosporioides）、葡萄白粉病菌（Uncinula necator）、葡萄灰霉病菌（Botrytis cinerea）、葡萄白腐病菌（Coniella diplodiella）、葡萄溃疡病菌（Botryosphaeria dothidea）、白菜黑斑病菌（Alternaria brassicae)、辣椒炭疽病菌（C. capsica）、甘草根腐病菌（Fusarium solani）、西葫芦白粉病菌（Sphaerotheca fuliginea）、番茄菌核病菌（Sclerotinia sclerotiorum）、马铃薯干腐病菌（F. equiseti）、西瓜枯萎病菌（F. oxysporum）、哈茨木霉（Trichoderma harzianum）等14种葡萄及其他作物病原菌和拮抗菌的菌丝DNA进行常规PCR和real-time PCR特异性检测,并对灵敏度和可重复性进行评价。运用已构建的real-time PCR体系对人工接种葡萄霜霉病菌的潜育期叶片内病原菌DNA进行定量检测,利用SPSS 19.0软件分析接种时间与叶片内葡萄霜霉病菌潜伏侵染量的关系。【结果】研究设计的引物特异性高,常规PCR仅对葡萄霜霉病菌DNA有扩增条带,为139 bp;real-time PCR检测结果表明该对引物对葡萄霜霉病菌有唯一的产物吸收峰,对其他供试菌株均未检测到产物吸收峰。常规PCR检测的灵敏度为10 pg·μL ^-1,real-time PCR的灵敏度可达到0.1 pg·μL ^-1,是常规PCR检测灵敏度的100倍。以携带目的基因片段的重组质粒为标准品,构建real-time PCR循环阈值（Ct）与模板浓度的线性关系,标准曲线y=42.27-3.36x,相关系数R ²=0.997,扩增效率为98.50%,线性范围达7个数量级,在2.4×10 ³—2.4×10 ⁹ copies/μL呈现良好的线性关系。对人工接种葡萄霜霉病菌的潜育期叶片内病原菌DNA进行real-time PCR检测,结果表明叶片内病原菌潜伏侵染量随接种时间的变化呈指数关系增长,y=6.34×10 ⁴·e ^{0.084 x},相关系数R ²=0.936。该real-time PCR检测体系在接种6 h后就可以检测到葡萄霜霉病菌DNA,检测量为5.68×10 ⁴ copies/μL病原菌DNA。 【结论】构建的葡萄霜霉病菌real-time PCR检测体系的灵敏度远高于常规PCR,且特异性强、重复性好,其Ct值与模板浓度呈较好的线性关系,扩增效率高,可用于定量检测葡萄霜霉病菌的潜伏侵染量。     

An improved method for preparing Agrobacterium cells that simplifies the Arabidopsis transformation protocol

Background  

The Agrobacterium vacuum (Bechtold et al 1993) and floral-dip (Clough and Bent 1998) are very efficient methods for generating transgenic Arabidopsis plants. These methods allow plant transformation without the need for tissue culture. Large volumes of bacterial cultures grown in liquid media are necessary for both of these transformation methods. This limits the number of transformations that can be done at a given time due to the need for expensive large shakers and limited space on them. Additionally, the bacterial colonies derived from solid media necessary for starting these liquid cultures often fail to grow in such large volumes. Therefore the optimum stage of plant material for transformation is often missed and new plant material needs to be grown.     

一株鹅星状病毒变异毒株的基因组特征与致病性分析

旨在研究引起雏鹅痛风的新型鹅星状病毒流行株特征。本试验从安徽省鹅星状病毒阳性雏鹅组织中进行病毒分离,并对分离的毒株进行全基因组测序,随后进行遗传进化分析。结果表明,病毒液接种9~11日龄鹅胚可成功分离到病毒,将其命名为GASTV-ZM株。IFA （indirect immunofluorescence assay）鉴定为阳性。基因组序列分析表明,ZM株鹅星状病毒基因组全长7 175 nt,ORF2基因核苷酸和氨基酸与2014—2020年间分离毒株的同源性分别为96.7%~98.9%和96.9%~99.0%,ORF2基因编码的氨基酸序列结果显示,3个氨基酸突变位点与以往存在显著差异,ZM株与2016年以来在我国流行的新型鹅星状病毒处于同一进化分支,与2020分离株HNSQ-6株和XT1株同源性较高。本研究为进一步研究该病毒的致病性和致病机理奠定基础。     

Differences in chemical composition relative to functional differentiation between petioles and laminas of Fraxinus excelsior

Differences in structural and nonstructural carbohydrates, lignin and chlorophyll, and Rubisco (ribulose-1,5-bisphosphate carboxylase/oxygenase) activity between petioles and leaflets were studied along a canopy light gradient in Fraxinus excelsior L., which has pinnate compound leaves and up to 20% of foliar biomass invested in petioles. Long-term light conditions at the sampling locations were characterized by values of seasonal mean integrated quantum flux density (Q(int), mol m(-2) day(-1)) estimated by combining data from hemispherical photographs at the sampling locations with measurements of global solar radiation above the canopy during the growing season. The contribution of petioles to leaf carbon assimilation was disproportionally lower than that of leaf laminas. Though the light relationships of assimilative compounds-foliar chlorophyll concentration increasing with decreasing Q(int) to improve leaf absorptance, foliar N concentration and Rubisco activity being relatively constant along the light gradient-were similar for both petioles and leaflets, petiole nitrogen and chlorophyll concentrations were only 30% and 10%, respectively, of those of leaflets. Nonstructural carbohydrate concentration was about 20% higher in petioles than in leaf laminas, indicating that petioles also serve as storage tissues for photosynthates. Relationships between foliar structural carbon components and irradiance-increasing lignin (L) and decreasing structural polysaccharide (SP) concentrations with increasing Q(int)-were qualitatively similar for petioles and leaflets. However, petioles had lower L, but higher SP and total investment in structural compounds (L + SP) than leaflets. Greater lignification at high irradiances in leaflets than in petioles was attributed to greater water stresses at high light, and to more variable water contents of actively transpiring leaflets. Low lignin concentration in combination with high osmotically active carbohydrate concentrations in petioles suggest that turgor plays an important role in the mechanical properties of petioles. As a result of lower lignin and protein concentrations, the glucose cost of petiole construction (g glucose per g dry mass) was about 5% lower than that of leaf laminas.     

Species differences in timing of leaf fall and foliage chemistry modify nutrient resorption efficiency in deciduous temperate forest stands

Extensive variation in fractional resorption of mineral elements from plant leaves is still not fully understood. In multi-species forest stands, species leaf fall phenology and leaf constitution may significantly modify the timing of nutrient return to the soil and overall plant nutrient loss. We studied leaf fall and nutrient loss kinetics, and leaf composition in three natural, temperate, deciduous broadleaf forest stands to determine the role of timing of leaf abscission and nutrient immobilization in cell walls on nutrient resorption efficiency of senescing leaves. Nitrogen (N), phosphorus and potassium contents decreased continuously in attached leaves after peak physiological activity during mid-season. Changes in nutrient contents of attached leaves were paralleled by decreases in nutrient contents in freshly fallen leaf litter. In different species and for different nutrients, resorption of nutrients from senescing leaves proceeded with different kinetics. The maximum nutrient resorption efficiency (the fraction of specific nutrient resorbed from the leaves at the end of leaf fall) did not depend on the mid-seasonal nutrient concentration. Species with earlier leaf fall resorbed leaf nutrients at a faster rate, partly compensating for the earlier leaf fall. Nevertheless, the litter-mass weighted mean nutrient contents in leaf litter were still larger in species with earlier leaf fall, demonstrating an inherent trade-off between early leaf fall and efficient nutrient resorption. This trade-off was most important for N. Losses of the non-mobile nutrients calcium and magnesium were unaffected by the timing of leaf fall. There was large variation in the maximum N resorption efficiency among species. Correlations among leaf chemical variables suggested that the maximum N resorption efficiency decreased with the increasing fraction of cell walls in the leaves, possibly due to a greater fraction of N occluded in cell wall matrix. We conclude that species leaf fall phenology and leaf chemistry modify the timing and quantities of plant nutrient losses, and that more diverse forest stands supporting a spectrum of species with different phenologies and leaf types produce litter with more variable chemical characteristics than monotypic stands.     

Perforation hyphae of soft rot fungi in the wood of white spruce [Picea glauca (Moench.) Voss.]

Summary Observations made on ponded white spruce wood with the electron microscope revealed that initial branching of perforation hyphae of soft rot fungi did not parallel microfibrillar orientation in the S₂ wall layer but rather occurred perpendicular to this alignment. In the absence of structural evidence suggesting mechanical entry it appeared that cell wall pencetration was accomplished through enzymatic activity at the hyphal apex.The authors wish to express their thanks to Miss S. Godkin and Mr. G. Chauret for technical assistance.