解磷细菌 K3 的 GFP 标记及其解磷能力检测

本文运用构建成功的含有假单胞菌属自身启动子及绿色荧光蛋白(GFP)的质粒pTRGFP电转至假单胞菌解磷细菌K3中,通过激光共聚焦显微镜及质粒检测,获得了发光稳定的标记菌株K3GFP。7天液体摇瓶试验中,标记菌株K3GFP的可溶性P含量在第4天达到最高774μg/ml,而出发菌株K3在第3天时已经达到了最大含量780μg/ml,说明pTRGFP质粒的转入对K3菌株解P能力有一定的影响。标记菌株K3GFP施入自然土壤10天后数量维持在5.47×106CFU/g～2.40×106CFU/g,35天后降到5.0×103CFU/g,说明解磷细菌K3GFP可以在自然土壤中定殖。本试验为研究解磷细菌在根际及土壤中的生长动态等行为特征奠定了基础。     

钩状木霉 ACCC31649的GFP标记及其对辣椒定殖和促生作用

【目的】旨在建立农杆菌介导的绿色荧光蛋白 (GFP) 基因转化钩状木霉 (Trichoderma hamatum) ACCC31649的技术方法,筛选遗传稳定的GFP标记转化子,并研究该菌株在辣椒植株中定殖和对辣椒的促生作用,为进一步阐明木霉在辣椒根际定殖及其与辣椒病原菌在辣椒根际和植株中的定殖、互作和生防作用奠定基础。【方法】通过根癌农杆菌介导的遗传转化方法筛选遗传稳定的GFP标记转化子,通过灌根接种方法和组织切片的水玻片荧光观察研究了钩状木霉在辣椒植株中定殖过程和对辣椒的促生作用。【结果】获得了遗传稳定GFP标记的钩状木霉转化子。荧光显微观察表明,辣椒根、茎、叶组织中都检测到GFP标记菌株的定殖。标记菌株首先在根部定殖,然后通过根部维管束逐步定殖到茎和叶片组织中。野生型菌株和GFP标记菌株灌根接种4叶期辣椒幼苗,30天后,GFP标记菌株与水处理对照相比,辣椒的株高增长13.5%,根长增长16.2%,鲜重和干重分别增加了43.8%和45.3%,而且野生型菌株与GFP标记菌株对辣椒的促生作用没有显著差异。【结论】钩状木霉能够在辣椒植株根、茎和叶组织中定殖,并且对辣椒具有显著的促生作用。同时,GFP标记的钩状木霉将在进一步阐明该菌株在辣椒根际定殖及其对病原菌拮抗和互作研究中发挥重要作用。     

缩二脲降解菌在作物根际的GFP标记示踪

通过三亲结合对缩二脲降解菌GW-1菌株成功进行了绿色荧光蛋白（Green fluorescent protein, GFP）基因标记,标记菌株命名为GW-1-GFP。实验证明外源质粒对宿主菌的生长未带来不利的影响,且经高效液相色谱测定该标记菌株降解缩二脲的能力与出发菌株无显著差异（p<0.05）。经抗生素抗性和荧光追踪证明标记菌株GW-1-GFP能够很好地在土壤中定殖,第25天时在不同处理的土壤中标记菌株对缩二脲的降解能力都超过50%,45天后该标记菌株在土壤中已难以检测。经标记菌株GW-1-GFP菌悬液浸润的小麦种子发芽后,通过荧光观察显示降解缩二脲的标记菌株GW-1-GFP在小麦根部定殖良好,且该标记菌株能在一定程度上缓解缩二脲对小麦的毒害作用。该研究为验证、追踪土壤中功能微生物菌剂与作物根部的亲和性和生态有效性提供了简易、直观的检测方法。     

根际促生菌防控土传病害的机理与应用进展

根际促生菌作为一类活跃于植物根际的有益微生物,对抑制土传病害的发生,改善和维护土壤生态质量具有重要作用以及广阔的发展前景。本文综述了根际促生菌直接或间接的土传病害防控机理,包括产抗生素、产水解酶、释放挥发性气体、诱导抗性、分泌铁载体、分泌激素和固氮解磷等;亦从信号识别与迁移、定殖规律以及定殖的微生物群落影响等方面对根际促生菌的定殖防病机理予以了总结;此外,笔者还就根际促生菌的应用现状进行了概述。     

甲基对硫磷降解菌DLLBR在青菜及根际土壤中的定殖研究

将 GFP 基因标记的甲基对硫磷降解菌 DLLBR 接入 100 ml LB 培养基,过夜培养达到 1010cfu/ml,浇灌到盆钵试验的 800 g 土壤中,20 天后用激光共聚焦显微镜检测其在小青菜植株根部和植株内的定殖及分布。结果表明标记菌株能够在植株根圈较好地定殖,也能在植株内定殖。30 天后将植株各段研磨破碎,涂布 LB 平板计数发光菌落数,在根内的定殖数为 104cfu/g 根,在茎内的定殖数为 102cfu/g 茎。结果还发现与不接菌的对照相比,处理促进了植株内细菌群落结构的变化,尤其是芽孢杆菌的种类和数量明显增加。接菌 45 天后通过土壤计数检测发现,标记菌株在土壤中的存活力很高,能达到 106cfu/g 土,同时也发现与对照相比,接菌的土壤细菌群落结构明显发生了变化,细菌种类变化不大,但芽孢杆菌的数量明显增加。     

短小芽孢杆菌BX-4抗生素标记及定殖效果研究

为研究短小芽孢杆菌BX-4在作物根际的定殖及防病效果,通过浓度梯度法用氨苄青霉素对其进行了抗性标记,并通过番茄盆栽试验研究了其在根际土壤中的定殖规律。结果表明,筛选出的突变体菌株BX-4＇能够耐受浓度为200 μg·mL^-1的氨苄青霉素,并且具有耐药和遗传双重稳定性;应用试验显示该突变体菌株能成功在番茄根际定殖,接种20 d后根际土壤中存活数量达到最高值1.34×10^8 cfu·g^-1干土,以后逐渐下降,到50 d时趋于稳定;筛选的突变体菌株对番茄青枯病具有明显的防治效果,防效达37.9%-50.9%。短小芽孢杆菌BX-4在作物根部的定殖规律为揭示其生防机理及应用该菌提供了科学根据。     

GFP标记的植物促生菌B96-Ⅱ-gfp的定殖能力研究

采用绿色荧光蛋白基因标记技术研究了植物促生菌B96-Ⅱ的标记菌B96-Ⅱ-gfp在盆栽黄瓜土壤中的时间、空间定殖动态以及在黄瓜植株上的分布。研究表明: B96-Ⅱ-gfp在土壤中具有持久的定殖能力, 接种180 d时在自然土和黄瓜枯萎菌病土中的定殖数量分别为2.7×10⁴ cfu·g^-1和6.6×10⁴ cfu·g^-1, 360 d时在土壤中仍可检测到B96-Ⅱ-gfp的存在。B96-Ⅱ-gfp可在盆栽植株生长土壤的表层(0~4 cm)、中层(4~8 cm)和底层(8~12 cm)定殖, 定殖数量随土壤深度的增加而增加。此外, B96-Ⅱ-gfp还可在黄瓜的根、茎和叶上定殖, 根部定殖的数量(7.2×10⁴ cfu·g^-1)显著高于茎部和叶部定殖的数量; 黄瓜植株体内定殖的数量多于体表定殖的数量。对土壤中可培养的3大微生物类群影响的研究表明: B96-Ⅱ-gfp对土壤中真菌数量具有显著抑制作用, 而对细菌和放线菌数量则没有明显影响。防病促生试验表明: B96-Ⅱ-gfp可增加黄瓜株高、鲜重和干重, 对黄瓜枯萎病有一定的防治效果, 且与未标记菌株B96-Ⅱ的防病促生作用无显著差异。     

东北地区四种农作物根际磷细菌的分布

从小麦、水稻、玉米,大豆根际土壤中磷细菌数量及其群落结构、多样性和相似性等几个方面,对东北地区代表性农作物根际磷细菌生态分布规律进行了系统研究.结果表明,4种作物根际土壤中分布大量解磷菌和溶磷菌,其中解磷菌达到2.15～6.68×105cfu·g-1、溶磷菌达到0.7～3.9×104cfu·g-1,解磷菌数量多于溶磷菌;作物根际土壤有效磷含量与磷细菌数量问无直接相关性;东北地区农作物根际磷细菌种群结构复杂,小麦、玉米、水稻、大豆根际土壤解磷菌分别涉及9个、11个、13个、12个菌属,溶磷菌分别涉及9个、8个、9个、7个菌属:4种主要农作物根际土壤中分布的磷细菌具有较高的多样性指数,且一般来说根际土壤中解磷菌多样性更高;作物根际磷细菌组成相似性系数在0.50～1.00之间,总体处于较高水平,且溶磷菌的相似性系数高于解磷菌.     

内生菌YN201728的定殖能力及其防治烟草白粉病的效果研究

为研究生防菌YN201728在烟草体内的定殖规律及防病机制,本研究利用其绿色荧光标记菌株YN28-P43GFPmut3a实时动态观测标记菌的定殖部位及密度,并探究其对温室烟草白粉病的盆栽防效与定殖的关系。结果表明,YN28-P43GFPmut3a发酵液处理烟草种子和幼苗后,种子内的标记菌含量可达2.18×10⁶ CFU·g^-1,在幼苗的根表土、根际土、根、茎、叶等组织中均能检测到标记菌,且其定殖密度表现为根表土>根际土>根>茎>叶。激光共聚焦显微镜观察显示,标记菌主要聚集在烟草根表皮、木质部导管、茎表皮、韧皮部及维管束组织、叶片表面和叶肉细胞间隙以及种皮与胚等部位。盆栽防效试验结果表明,YN201728野生型和标记菌株对烟草白粉病均有较好的保护和治疗效果,持效期长达21 d。此外,烟草叶片中内生菌的定殖量与其防效呈正相关。本研究结果表明,内生菌YN201728在烟草体内有良好的定殖和防治白粉病效果,具很好的开发潜力,为烟草病害的生物防治提供了一定的理论基础。     

土壤因子对链霉菌S506定殖和促生功能的影响

链霉菌S506是从植物根际分离获得的具有促生、防病和降解根系毒素功能的根际微生物,为了解土壤环境条件对其在根际定殖和功能表达的影响,以链霉菌S506为试材,研究了土壤质地、环境温度和土壤湿度对S506在番茄根际土壤定殖及番茄植株生长的影响.结果表明,适宜链霉菌S506在番茄根际定殖和促生作用发挥的土壤质地为壤土,其次为砂土和黏土;试验所设温度梯度中,利于S506在根际定殖的环境温度为30℃,其次为22.5℃和15℃,而利于目的菌株促生功能表达的环境温度则依次为22.5℃、30℃和15℃;适宜于目的菌株在根际定殖和促生功能发挥的土壤相对湿度为20%、25%,其次为15%和30%.     

Silicon Effect on Nutrient Acquisition of Peanut (Arachis hypogaea L.) Under Aluminum Stress

Aluminum (Al) toxicity represents one of the main yield-limiting factors for crops in acid soils. Silicon (Si) is known to increase tolerance in higher plants. This study was conducted to determine whether treatment with Si could improve nutrient uptake by peanut under Al stress. Peanut (Arachis hypogaea L. cv Zhonghua 4) was raised with or without Si (1.5 mM) in the growth chamber under 0 and toxic Al (0.3 mM) levels. Aluminum stress significantly decreased the root- and total-dry weight by 52.4% and 32.0%, respectively. The content of nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), and magnesium (Mg) was significantly decreased, but that of Al increased markedly in shoots and roots after Al exposure at seedling, flower-needle, and pod-setting stage. Silicon alleviates Al toxicity in peanut plants in relation to Al distribution and allocation of tissue P, K, Ca, and Mg by favoring the partitioning of dry mass to roots.     

Trace Metal Contamination During Grinding of Plant Samples

Inductively coupled plasma-mass spectroscopic (ICP-MS) analysis of leaves from 22 cabbage crops in the Sa P? and B?c Hà districts of Láo Cai Province, North-Western Vi?t Nam, revealed unexpectedly high concentrations of chromium (Cr) and nickel (Ni). The concentrations were strongly linearly related (r² = 0.94), indicating sample contamination during grinding through a stainless-steel hammer mill. We tested this hypothesis in two ways. First, brown rice ground through the same mill was contaminated not only by Cr and Ni, but also cobalt (Co), iron (Fe) and molybdenum (Mo). Second, scanning electron microscopy/energy dispersive analysis of x-rays (SEM/EDS) of the ground samples revealed small fragments with co-located Fe, Cr and Ni, consistent with stainless steel wear fragments. Other grinders may perform differently and we suggest that quality assurance protocols for trace metal analysis of plants should include testing for grinder wear metals. Lastly, brown rice appears to be convenient for investigating contamination of plant tissues during grinding.     

模拟降雨条件下污染土壤中重金属元素径流迁移特征

探明污染土壤中重金属元素随地表径流迁移特征,对于防控重金属污染物的迁移扩散具有重要意义。通过室内模拟降雨试验,研究不同降雨强度下(55,100,120mm/h)污染土壤中Cd、Pb和Cr随地表径流迁移的特征。结果表明:径流中3种重金属元素总量在产流的前20min内逐渐降低,随后趋于稳定;产流初期(产流10min内)颗粒态重金属占径流重金属总量的比例均在80%以上;随产流时间延长,颗粒态Cd和Cr对径流重金属的贡献率快速降低,其颗粒态/溶解态的比值范围分别为47.0～0.4和12.9～0.4,而Pb则主要以颗粒态的形式存在,溶解态Pb对其随径流迁移的贡献率可忽略不计;随降雨强度增大,径流中总Cd和Pb含量显著降低,而径流中总Cr含量以及溶解态重金属含量无明显变化规律。     

Effects of humic acid on remediation of the nutritional deficiency of gerbera in hydroponic culture

Information is scant on the effect of humic acid (HA) on physiological, antioxidant and photosynthesis attributes of gerbera plants undergoing nutrient deficiency in culture solution. Gerbera plants cv. Malibu were grown in a factorial experiment based on a completely randomized design with 3 replications, using 3 different nutrient solutions [complete nutrient solution (NSc), 25% NSc (NS1), and 50% NSc (NS2)] treated with 2 levels of humic acid [0 (HA0) and 500 mg/l (HA1)].The interaction effect of HA and NS showed that HA improved the flower number in NSc, the transpiration in NS1+HA1, photosynthesis rate in NSc+HA1, stomatal conductance (gs) in NS2, mesophyll conductance of leaves in all NS levels and photosynthetic water use efficiency in NSc+HA1. The interaction effect of nutrient solution and HA on antioxidant activity was inconclusive, malondialdehyde content was the highest in NS2 and the lowest in NS1+HA1. The peroxidase activity increased in complete nutrient solution with and without HA and there were no differences among other treatments. Superoxide dismutase activity increased in NS1 and complete nutrient solution with HA and reached the highest in NSc. Humic acid was more effective in nutrient uptake, i.e., nitrogen, phosphorus, potassium, calcium, zinc, and iron (N, P, K, Ca, Zn, and Fe) in complete nutrient solution compared to NS1 and NS2. Conclusively, humic acid can compensate the nutrient deficiency stress of the culture solution in regards to protein synthesis, photosynthesis attributes regardless of the nutrient uptake of gerbera.     

Irrigation Module and Sowing Date Affect Seed Cotton Yield,Quality, Productivity Indices,and Economics of Cotton in North-western India

ABSTRACT

Delayed sowing and imprecise application of irrigation water to cotton has been the major hurdle in sustaining cotton yield in north-western India. Therefore, studies were initiated to ascertain the impact of heavy or normal level of presowing irrigation (PSI), scheduling time of first postsowing irrigation (POSI) under two sowing dates (SDs) on cotton arranged in a split block design replicated thrice. PSI_h (100 mm) recorded 23.2% higher seed cotton yield over PSI_n (70 mm) owing to better yield attributes and higher stand. First POSI at 4 weeks after sowing (WAS) resulted the highest yield (3072 kg ha^?1), while one at 6 WAS (POSI₆) recorded least due to poor population and reduced yield attributes. Water productivity under POSI₆ was lesser by 29.1%, 25.2%, and 16.2% as compared to POSI₃, POSI₄, and POSI₅, respectively. April sown crop out yielded the May sown cotton crop by 939 kg ha^?1 due to better yield attributes. Nitrogen factor productivity (NFP) among SD remained higher by 23.3% for April as compared to May sowing. PSI_h exhibited better NFP over PSI_n. POSI₆ recorded least NFP by 24.4%, 28.5%, and 16.3% as compared to POSI₃, POSI₄, and POSI₅, respectively and was indicative of relatively poor utilization of N under delayed schedules. Therefore, planting in April after heavy PSI and scheduling first POSI at 4 WAS is best strategy for sustaining cotton yield and maximizing farmer profitability.     

氮磷钾不同配比对饲用稻威优198产量及糙米蛋白含量的影响

采用田间小区试验研究“推荐配比”(N:P2O5:K2O比率为190:90:100 kg hm-2)、“高氮量配比”(N:P2O5:K2O比率为210:90:100 kg hm-2)、“低氮量配比”(N:P2O5:K2O比率为170:90:100 kg hm-2)以及“常规配比”(N:P2O5:K2O比率为216:112.5:202.5 kg hm-2)4种氮、磷、钾配比施肥对饲用稻威优198蔗糖合成酶(SUS)、腺苷二磷酸焦磷酸化酶(AGPase)、硝酸还原酶(NR)、谷氨酰胺合成酶(GS)以及产量和糙米蛋白质的影响。结果表明:“推荐配比”能提高不同生育时期水稻功能叶(旗叶)和粒籽中碳、氮代谢关键酶的活性,这些关键酶活性的变化显著影响水稻产量和糙米全氮以及蛋白氮的含量。统计(P0.05)结果证实“推荐配比”能提高水稻产量达到8200 kg hm-2,与“常规配比”相比产量提高了24.81%;“推荐配比”糙米全氮和蛋白氮含量分别达到22.70 g kg-1和21.98 g kg-1,与“常规配比”相比差异显著,并且其全氮和蛋白氮含量分别提高17.01%,18.38%。     

Arbuscular Mycorrhizal Fungi Enhance Tolerance of Rosa multiflora cv. Burr to Bicarbonate in Irrigation Water

ABSTRACT

High bicarbonate (HCO₃ ^?) of irrigation water can be detrimental to plant growth in sustainable horticultural production systems. The ability of arbuscular mycorrhizal fungi (AMF), ZAC-19, (composed of Glomus albidum, Glomus claroideum, and Glomus diaphanum) to enhance tolerance to HCO₃ ^? was tested on Rosa multiflora cv. Burr. Arbuscular mycorrhizal colonized and non-inoculated (non-AMF) plants were treated with 0, 2.5, 5, and 10 mM HCO₃ ^?. Increasing HCO₃ ^? concentration and associated high pH and electrical conductivity (EC)—reduced plant growth, nutrient uptake, and acid phosphatase activity, while increasing alkaline phosphatase activity (ALP). Inoculation with AMF enhanced plant tolerance to HCO₃ ^?, as indicated by greater growth (leaf, stem, and total plant dry weight, leaf area and leaf area ratio), leaf elemental concentration [nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), iron (Fe), zinc (Zn), aluminum (Al), boron (B)], leaf chlorophyll concentration, higher mycorrhizal inoculation effect, lower root Fe reductase activity, and generally lower soluble ALP activity. While AMF colonization was reduced by increasing HCO₃ ^? concentration, colonization still occurred at high HCO₃ ^? concentration. At 2.5 mM HCO₃ ^?, AMF plant growth was comparable to plants at 0 mM HCO₃ ^?, further indicating the beneficial effect of AMF for alleviation of HCO₃ ^? plant stress.     

NH4+-K+ co-loaded clinoptilolite as a binary fertilizer

ABSTRACT

Understanding the potential of clinoptilolite (CLI) for adsorption of NH₄⁺ and K⁺, providing appropriate fertilizer formula, and evaluation of the produced zeolitic nutrient sources (ZNSs) to meet the plant need are the main objectives of this study. Three ZNSs (NH₄⁺-saturated, K⁺-saturated and dual-purpose NH₄⁺-K⁺ saturated CLI) were produced, assessed, and compared with commercial N and K fertilizers (CFs) on corn growth in a greenhouse. The results indicated that CLI can potentially adsorb both NH₄⁺ and K⁺ to the maximum values of 25.00 mg-NH₄⁺ g^?1 and 47.61 mg-K g^?1, respectively, and chemisorptions mainly followed the process of adsorption. Saturation of zeolite by NH₄⁺ and K⁺ occurred after 10 and 15 d which lead to ZNSs with 2% and 5% of N and K, respectively. NH₄⁺-K⁺ saturated CLI contained 1% N and 1% K. The greenhouse experiment showed no significant difference between ZNSs and CFs on plant growth. However, the application of both N and K in the form of zeolitic sources significantly increased the uptake of N by the plant. The highest uptake of K (2.05 g pot^?1) occurred in plants supplied with both natural zeolite and CFs. The present results may benefit the future utilization of ZNSs in environmental friendly farming practices.     

四种常规方法提取伊利石有效钾的机制比较

采用化学分析、X射线衍射、中红外光声光谱以及原子力显微镜的方法,比较了0.2 mol L~(-1)四苯硼钠法、1 mol L~(-1)沸硝酸法、2 mol L~(-1)冷硝酸法和2 mol L~(-1)热盐酸法浸提伊利石中有效钾的机制。结果表明,四苯硼钠法浸提时,伊利石中钾素释放量达到全钾量的59.5%,且基本均通过层间交换反应予以释放,结构离子铁、铝和硅释放量极低;采用三种酸溶液浸提时,其钾素释放量仅占全钾量的1.53%~2.46%,通过层间交换反应释放的钾量占释放量的比例为88.4%~94.0%。四苯硼钠浸提时伊利石层间距扩大,产生次生过渡矿物,并形成富硅表层,但在伊利石表面无溶蚀特征;三种酸溶液浸提时伊利石结构无改变,但其结晶度降低,且表面有明显的溶蚀特征。因此,土壤矿物层间钾是作物可利用有效钾的主要来源,三种酸溶液浸提方法一方面低估了有效钾容量,另一方面提取了一部分不能为植物所利用的结构态钾,不适宜于用来评价伊利石及土壤有效钾库容量。     

氨甲喋呤(MTX)的直接碘标记方法研究

本文报道了~125I(~131I)抗肿瘤药物氨甲喋呤的制备方法,用直接碘化法——NBS、ChT、Iodogen进行标记,并对这几种方法进行了比较。结果表明,NBS法最好,标记率达90—99％。经Sephadex G-75柱凝胶过滤后得到纯产品,放化纯度为95.4％。所制得的标记产品适用于示踪研究。