玉米根内和根际土壤溶磷细菌生物学特性研究

从玉米根内和根际土壤中分离筛选到22株溶磷细菌。根据16SrRNA基因序列分析,2株溶磷效果最好的菌株分别被鉴定为伯克霍尔德氏菌(Burkholderia sp.SY9)和泛菌(Pantoea sp.Gym7c)。同时对菌株SY9和Gym7c的溶磷能力和植物促生特性进行了进一步研究。利用磷酸钙和开阳磷矿矿粉研究了菌株释放P、Ca和Fe的效能。液体培养条件下,菌株在在磷酸钙培养基中的生长好于在开阳磷矿粉培养基中的生长,同时菌株能够合成铁载体。菌株SY9从开阳磷矿粉中释放P、Ca,和Fe的能力比菌株Gym7c强。然而菌株Gym7c从磷酸钙中释放P和Ca的能力比菌株SY9强。菌株Gym7c主要是通过合成的有机酸溶解磷酸钙并释放其中的P和Ca,而菌株SY9是通过合成的有机酸和铁载体溶解开阳磷矿粉并释放出其中的P、Ca,和Fe。溶解开阳磷矿粉的有机酸主要是葡萄糖酸而溶解磷酸钙的有机酸主要包括丁二酸、丙酸、苹果酸和酒石酸等。菌株Gym7c能够合成生长素、铁载体和1-氨基环丙烷-1-羧酸脱氨酶。菌株SY9和Gym7c对酸碱、盐和温度均表现出一定的抗性。     

生活垃圾堆肥对难溶性磷有效性的影响

生活垃圾在堆肥过程中将产生有机酸类物质,这些酸性物质对难溶性磷具有较强的溶解能力.本研究在生活垃圾堆肥过程中,加入难溶性磷矿粉,系统探讨堆肥对难溶性磷的转化及堆肥产品对磷素有效性的影响.研究表明,速效磷在堆肥中后期达到最大值,并在堆肥的后期相对稳定,堆肥结束后,扣除磷矿粉本身含有的速效磷对堆肥的影响,与不加磷矿粉堆肥相比,P₁、P₂堆肥处理方式速效磷含量增加值分别为0.87 g·kg^-1、0.76 g·kg^-1.电镜观察表明,堆肥结束后,磷矿粉表面棱角消失,呈蜂窝状.堆肥产品培肥后,富磷垃圾肥在作物生育期内,土壤速效磷含量、酸性磷酸酶活性均高于垃圾肥、腐熟鸡粪及化肥.     

一株穗花狐尾藻内生菌的分离鉴定及其溶磷特征研究

通过组织表面消毒、切面培养和共生培养的方法从健康的穗花狐尾藻植株体内分离获得了一株具有较强溶磷能力的内生菌,系统发育分析和VITEK 2系统鉴定结果表明,内生菌与沙雷氏菌的亲缘关系最近,因此,命名为Serratia sp.SP5.纯培养实验表明,菌株SP5通过产生酸性物质溶磷,在培养96 h后溶磷能力达到最大,溶解性磷酸根的浓度高达269.60 mg·L-1,培养液的p H值下降至4.32.氮源通过降低菌株SP5的产酸能力而间接影响了其对磷的释放能力,菌株SP5溶磷的氮源优先利用顺序为铵根离子硝酸根离子尿素.适宜菌株SP5生长的p H值范围为5~9,酸性条件下菌株的溶磷能力大于碱性条件,而在p H值为7时与对照相比菌株对磷的相对释放量最大.p H值达到9时,菌株无明显溶磷能力.共存高浓度的金属离子时菌株SP5的溶磷效果明显降低,影响程度的大小顺序为MnFeMg.菌株SP5溶磷的最佳碳氮比为35∶1,在相同碳氮比下,不同的碳氮量会明显影响溶磷效果.底泥模拟培养条件下,菌株SP5可通过释放底泥中的闭蓄态磷(O-P)和钙磷(Ca-P)而明显提高水相中的磷酸根离子浓度,与对照相比,底泥中这两种磷形态分别下降了15.21%和34.87%,而水相中溶解性磷酸根离子是对照的1.5~2.0倍.研究结果为研究富营养化水体中磷的生物地球化学过程提供了新的科学数据.     

Increased acidification in the rhizosphere of cactus seedlings induced by Azospirillum brasilense

Acidification of the rhizosphere of cactus seedlings (giant cardon, Pachycereus pringlei) after inoculation with the plant growth-promoting bacterium Azospirillum brasilense Cd, in the presence or absence of ammonium and nitrate, was studied to understand how to increase growth of cardon seedlings in poor desert soils. While ammonium enhanced rhizosphere and liquid culture acidification, inoculation with the bacteria enhanced it further. On the other hand, nitrate increased pH of the rhizosphere, but combined with the bacterial inoculation, increase in pH was significantly smaller. Bacterial inoculation with ammonium enhanced plant growth.     

Accumulation fatty acids of in Chlorella vulgaris under heterotrophic conditions in relation to activity of acetyl-CoA carboxylase,temperature, and co-immobilization with Azospirillum brasilense

The relation between fatty acid accumulation, activity of acetyl-CoA carboxylase (ACC), and consequently lipid accumulation was studied in the microalgae Chlorella vulgaris co-immobilized with the plant growth-promoting bacterium Azospirillum brasilense under dark heterotrophic conditions with Na acetate as a carbon source. In C. vulgaris immobilized alone, cultivation experiments for 6 days showed that ACC activity is directly related to fatty acid accumulation, especially in the last 3 days. In co-immobilization experiments, A. brasilense exerted a significant positive effect over ACC activity, increased the quantity in all nine main fatty acids, increased total lipid accumulation in C. vulgaris, and mitigated negative effects of nonoptimal temperature for growth. No correlation between ACC activity and lipid accumulation in the cells was established for three different temperatures. This study demonstrated that the interaction between A. brasilense and C. vulgaris has a significant effect on fatty acid and lipid accumulation in the microalgae.     

攀枝花矿区糙野青茅根际耐铬放线菌筛选及促生能力评价

从攀枝花矿区糙野青茅根际土中分离放线菌,筛选获得对重金属铬具有强耐受性、抗菌促生和产胞外酶能力的放线菌,为微生物-植物联合修复重金属污染提供菌种资源.采用稀释涂布法进行放线菌的分离,通过放线菌在不同铬浓度高氏一号培养基上的生长状况判断其对重金属铬的耐受性,进一步采用原子吸收分光光度法分析菌株对铬耐受性强的菌株耐铬及去铬能力;通过对菌株产铁载体、溶磷、产吲哚乙酸(IAA)、产纤维素酶、产淀粉酶、产木糖酶、产酪蛋白酶能力及抗菌活性的筛选,综合评价菌株抗菌促生能力.结果显示,分离获得的27株根际菌株,光学显微形态及扫描电镜结果均显示为放线菌,其中有14株能在含铬的培养基上生长,具有耐铬能力.16S rRNA基因测序结果显示,14株放线菌分属于链霉菌属(streptomyces)、小单孢菌属(Micromonospora)和冢村氏菌属(Tsukamurella).菌株SCAU9002和SCAU9008在铬浓度为300mg·L~(-1)的处理下培养6 d,去铬率分别达到81.17%和78.41%;菌株SCAU9002、SCAU9006、SCAU9008、SCAU9009、SCAU9013同时具有3种促生功能,盆栽实验结果进一步表明,在重金属胁迫下,经菌株处理后的玉米植株更高,生物量更大,根系更发达,促生效果较好.此外,菌株SCAU9006对6种病原菌具有拮抗作用,5株菌同时具有3种及以上产胞外水解酶的能力,4株菌能在7%的NaCl浓度下生长,13株菌在pH5.0~8.0的范围内均可生长.结果表明,根际放线菌在促进矿区植物生长和铬尾矿污染土壤的生物修复研究中具有很大潜力.     

红树林土壤解磷菌的分离鉴定及解磷特性

从罗源湾红树林根际土壤中分离解磷菌,筛选出洋葱伯克霍尔德菌（Burkholderia cepacia, NR 113645.1）和短小芽孢杆菌（Bacillus pumilus, NR 043242.1）,研究它们的解磷特性和动态解磷过程. HPLC结果表明, B. cepacia菌液上清中含葡萄糖酸、丙酮酸、乳酸、乙酸、丁二酸, 这些有机酸的产生导致磷酸盐溶解. B.pumilus菌液上清中仅检测到少量的葡萄糖酸, 因此该菌溶磷效果不佳. 在细菌动态溶磷过程中发现, 经B. cepacia菌处理的磷酸钙, 其X射线衍射（XRD）特征峰强度随时间的增长降低, 显微镜图像显示磷酸钙颗粒随时间增长变小, 甚至消失. 而经B.pumilus菌处理的结果表明, 在试验期间内磷酸钙峰强度和颗粒大小没有明显变化. 细菌产生有机酸是溶解磷酸盐的重要前提, 这是解决板结土壤中难溶性磷源转化为生物可利用磷源问题的关键.     

一种修复Pb-Cd污染污泥的复合淋洗剂的研究

以某自来水厂污泥为背景泥样,人工添加重金属后泥样为研究对象,考察了葡萄糖酸、氯化钠、六偏磷酸钠复配溶液对污泥中重金属的洗脱效果.研究发现,复配淋洗剂对污泥中Pb和Cd有较好的萃取效果.形态分析表明,葡萄糖酸对污泥中的水溶态、离子交换态、碳酸盐结合态及部分弱有机结合态形式的重金属Pb、Cd有较好的萃取效果;六偏磷酸钠主要...     

Phosphatase activity and culture conditions of the yeast Candida mycoderma sp. and analysis of organic phosphorus hydrolysis ability

Orthophosphate is an essential but limiting macronutrient for plant growth. About 67% cropland in China lacks sufficient phosphorus, especially that with red soil. Extensive soil phosphorus reserves exist in the form of organic phosphorus, which is unavailable for root uptake unless hydrolyzed by secretory acid phosphatases. Thus, many microorganisms with the ability to produce phosphatase have been exploited. In this work, the activity of an extracellular acid phosphatase and yeast biomass from Candida mycoderma was measured under different culture conditions, such as pH, temperature, and carbon source. A maximal phosphatase activity of 8.47 × 10^5±0.11× 10^5 U/g was achieved by C. Mycoderma in 36 hr under the optimal conditions. The extracellular acid phosphatase has high activity over a wide pH tolerance range from 2.5 to S.0 （optimum pH 3.5）. The effects of different phosphorus compounds on the acid phosphatase production were also studied. The presence of phytin, lecithin or calcium phosphate reduced the phosphatase activity and biomass yield significantly. In addition, the pH of the culture medium was reduced significantly by lecithin. The efficiency of the strain in releasing orthophosphate from organic phosphorus was studied in red soil （used in planting trees） and rice soil （originating as red soil）. The available phosphorus content was increased by 230% after inoculating 20 days in rice soil and decreased by 50% after inoculating 10 days in red soil. This work indicates that the yeast strain C. mycoderma has potential application for enhancing phosphorus utilization in plants that grow in rice soil.     

利用浊度分析法研究硅酸对磷酸钙沉淀的影响

磷酸钙沉淀法是从污水中回收磷的最常用的方法之一.本文通过测定溶液浊度的方法研究了钙磷比为2∶1的条件下pH和硅酸对磷酸钙沉淀的影响.结果发现,pH低于6时,短时间内溶液中不能形成磷酸钙沉淀;而pH大于7时,溶液中迅速生成磷酸钙沉淀,且随pH的增加沉淀速度变大.XRD谱图表明,pH=7或8时都生成热力学最稳定的羟基磷酸钙沉淀.7 mg·L-1单硅酸的存在使生成的磷酸钙溶液的初始浊度从10NTU增加到20 NTU,使磷酸钙沉淀速度加倍,说明单硅酸能促进磷酸钙的沉淀速度.而聚硅酸使溶液初始浊度从10 NTU降低到0 NTU,抑制了磷酸钙的沉淀速度,表明不同形态的硅酸对磷酸钙的沉淀速度有不同的影响.XRD谱图表明,硅酸存在时,溶液中除了生成羟基磷酸钙外,还生成其他类型的磷酸钙.     

肠杆菌FM-1强化积雪草修复镉污染土壤机理

分析了肠杆菌FM-1的耐镉性及促生特性，并研究了肠杆菌FM-1对矿山型和非矿山型积雪草镉富集及根际土壤pH值的影响.结果表明，肠杆菌FM-1对镉有较强的耐受性，其分泌的吲哚乙酸含量，铁载体A/A_r值和溶磷能力分别达到（72.85±0.62）mg/L，0.21±0.01和（143.33±2.13）mg/L，表明肠杆菌FM-1具有良好的促生能力.将肠杆菌FM-1接种于采自广西柳州泗顶铅锌矿周边4种类型的土壤中，均不同程度提高了矿山型和非矿山型积雪草对镉的富集.其中，下游区土壤接种肠杆菌FM-1后，矿山型积雪草茎和叶中镉含量分别增加了87.90%~161.84%和21.85%~76.42%，非矿山型叶片中的镉含量增加了5.84%~35.20%.研究表明肠杆菌FM-1促进了积雪草对镉的富集.同时，肠杆菌FM-1的添加在一定程度上影响了积雪草根际土壤的pH值，显著降低了下游区矿山型积雪草根际土壤中的pH值.     

溶磷细菌筛选及在复垦土壤上对难溶态磷的活化作用

为研究溶磷细菌在复垦土壤上对难溶态磷的活化作用,通过PVK平板法从石灰性土壤中分离筛选出7株溶磷细菌,7株菌株的溶磷能力在296.5～563.5 mg/L之间,菌株的溶磷能力与溶磷圈直径（D）与菌落直径（d）的比值呈显著正相关（p<0.01）;16sRNA序列分析表明,W1、W6属于Enterobacter sp.,W2、W4属于Burkholderia sp.,W3、W5属于Rahnella sp.,W7属于Fluorescent pseudomonas。选择W3、W4、W7作为试验菌株,以磷酸三钙、磷矿粉为难溶态磷源,研究单菌株及组合菌株在配施难溶态磷酸盐条件下对复垦土壤有效磷、各形态无机磷含量及油菜产量的影响。结果表明,同一底物下溶磷细菌各处理复垦土壤有效磷含量和油菜产量显著高于基质处理（p<0.05）,组合菌株处理优于单菌株处理,W3+W4+W7处理两项指标都最高;在溶磷细菌+磷酸三钙底物下,W3+W4+W7处理复垦土壤有效磷含量和油菜产量分别比单施溶磷细菌、溶磷细菌+磷矿粉两个底物下同一处理显著增加94.81%、40.25%和34.13%、9.43%;相同底物下溶磷细菌各处理复垦土壤Ca₂-P、Ca₈-P、Al-P、Fe-P含量都高于基质处理,Ca₁₀-P含量小于基质处理,O-P含量与基质处理差异不显著。这一结果可为溶磷细菌在复垦土壤上应用技术的研发提供理论基础。     

Phosphorus release from phosphate rock and iron phosphate by low-molecular-weight organic acids

Low-molecular-weight(LMW) organic acids widely exist in soils, particularly in the rhizosphere. A series of batch experiments were carded out to investigate the phosphorus release from rock phosphate and iron phosphate by low-molecular-weight organic acids.Results showed that citdc acid had the highest capacity to solubilize P from both rock and iron phosphate. P solubUization from rock phosphate and iron phosphate resulted in net proton consumption. P release from rock phosphate was positively correlated with the pκa values. P release from iron phosphate was positively correlated with Fe-organic acid stability constants except for aromatic acids, but was not correlated with pκa. Increase in the concentrations of organic acids enhanced P solubilization from both rock and iron phosphate almost linearly. Addition of phenolic compounds further increased the P release from iron phosphate. Initial solution pH had much more substantial effect on P release from rock phosphate than from iron phosphate.     

不同磷肥对砷超富集植物蜈蚣草修复砷污染土壤的影响

施肥是提高重金属污染土壤的植物修复效率的必要强化措施之一.通过研究农业生产中常用磷肥对砷超富集植物蜈蚣草生长、砷累积及氮磷钾吸收的影响,确定可提高蜈蚣草修复砷污染土壤能力的最佳磷肥类型.结果表明,与不施砷对照组相比.添加砷处理中施用相应磷肥的蜈蚣草总生物量中蜈蚣草均有所降低(除钙镁磷肥外);在添加砷的情况下,钙镁磷肥、磷酸二氢钙和磷酸二氢铵等3个施磷处理总生物量相对较高.显著高于其他磷肥处理和不施磷处理(0.83 g/pot);地上部砷累积量由大到小的顺序为:磷酸二氢钙>钙镁磷肥>磷酸二氢铵>磷酸二氢钾>磷酸二氢钠>不施磷>过磷酸钙;磷酸二氢钙处理的砷去除效率高达7.28%.由此可见,磷肥能一定程度提高砷超富集植物的修复能力,但不同磷肥对其砷去除效率的影响可达数倍之差,磷酸二氢钙是首选磷肥类型,综合考虑土壤pH、N、P和有效态砷等各种因子,磷酸二氢铵和钙镁磷肥也可作备选肥料类型.     

化学沉淀法回收污泥中氮磷的影响因素研究

污水处理厂污泥上清液中富集着较高浓度溶解性的氮磷,将此部分氮磷形成磷酸盐沉淀(如磷酸氨镁、磷酸钙、磷酸铝等)加以回收利用,受到各种因素的影响.文章以正交试验得出的影响因素为基础,深入研究了pH、初始PO43--p的浓度、Mg/P和反应时间对某污水厂污泥上清液中磷酸氨镁沉淀法回收氮磷的影响.结果表明:pH是影响污泥上清液...     

多聚磷酸盐激酶基因在污水生物除磷中的功能

为验证多聚磷酸盐激酶基因(ppk)在污水生物除磷中的功能.采用Red敲除系统,以p KD4质粒为模板,设计同源短臂,扩增外源线性DNA片段,将外源线性DNA片段电转化整合入已导入p KD46的大肠杆菌ATCC25922野生型菌株.获得重组菌E.coli/ppk~-Kan~+.将p CP20导入大肠杆菌E.coli/ppk~-Kan~+以消除卡那霉素抗性基因,通过负抗性筛选及正反向引物验证,构建无抗生素抗性的ppk基因缺失工程菌株E.coli/ppk~-Kan~-.比较工程菌株和野生型菌株的生长特性,并比较两者在缺磷诱导/富磷及多次厌氧/好氧诱导条件下的除磷性能.结果表明采用Red重组系统,通过无痕敲除,成功构建了大肠杆菌ppk基因缺失菌株E.coli/ppk~-Kan~-.敲除后的工程菌株和野生型菌株生长整体没有差异,但是4 h前对数期工程菌株生长快于野生型菌株,8 h后稳定期工程菌株生长慢于野生型菌株,表明ppk影响菌体的生长;缺磷诱导/富磷条件下,工程菌株并未表现出因ppk缺失而影响其除磷能力;经过5次厌氧/好氧诱导,两菌菌体含磷量保持在1%~2%,没有因诱导次数的增加而表现出菌体含磷量增加的趋势,也未发现厌氧有PHB好氧有聚磷颗粒生成,表明ppk基因的缺失并没有引起菌体除磷能力的下降.ppk并未表现出明显的与污水生物除磷相关的功能.     

一株耐硝酸盐的巨大芽孢杆菌溶磷特性研究

以多种磷源培养基培养巨大芽孢杆菌(Bacillus megatherium)NCT-2,研究了该菌的溶磷特性.结果表明,在以磷酸钙为磷源时,溶磷效果随菌株的生长而发生变化,在第2 d菌株生长量最大,溶磷效果最好;不同碳氮源会影响菌株的溶磷效果,菌株优先利用葡萄糖,其次为麦芽糖和蔗糖,而对淀粉的利用能力较差;对氮源的利用顺序依次为(NH4)2SO4NH4Cl尿素Na NO2KNO3;磷酸钙的浓度会影响菌株溶磷效果,浓度过低或过高都会导致溶磷量过低.菌株对多种难溶性含磷物溶解效果的研究表明,菌株对植酸钙溶磷量最高,为98.1 mg·L-1,对Fe PO4、Al PO4的溶解能力稍弱,在30~40 mg·L-1之间,对卵磷脂溶磷量最小,为6.15 mg·L-1.通过对培养基中磷酸酶活性和有机酸进行分析,初步研究了NCT-2的溶磷机制,发现该菌通过产生酸性磷酸酶、碱性磷酸酶或产酸溶解难溶磷,不同磷源的溶磷机制有所不同.     

外源茉莉酸对水稻幼苗根系砷积累及抗逆应答效应

砷（As）污染对作物生长有毒害作用,导致作物品质和产量降低,因此亟需采取安全有效的策略以减轻其毒害.采用水培法,通过荧光探针定位技术、差速离心技术和傅里叶红外光谱技术（FTIR）研究外源茉莉酸（JA）对水稻幼苗As积累及抗逆应答的影响.结果表明,外源添加JA使水稻地下部和地上部的As含量分别降低31.4%和51.4%,显著降低细胞壁和细胞液中的As含量.JA改变亚细胞组分中As的分布比例,使细胞壁中As的分布比例提高16.4%,细胞液中As的分布比例降低17.3%,增强细胞壁对As的固定,减少细胞液中的As含量.此外,JA提高根细胞中SOD、CAT、GSH和PEPC的水平,降低H₂ O₂和MDA含量,减轻脂质过氧化损伤,调节碳氮代谢,缓解As毒害.研究为水稻As污染防治提供了新思路.     

软水中富里酸对铜管/磷酸盐体系溶解性铜释放影响

通过静态模拟试验研究了在软水体系中富里酸对铜管/磷酸盐缓蚀行为的影响,结果表明,富里酸含量、软水pH值、停留时间影响磷酸盐对铜管的缓蚀作用;高浓度的富里酸(16mg/L,以TOC计)明显增加溶解性铜释放,且从新铜管释放溶解性铜浓度高于老化6个月和12个月的铜管;在富里酸存在下,pH值较低时显著增加溶解性铜的释放,随着pH值的增大,富里酸对溶解性铜的释放影响降低;在96h的停留过程中,溶解性铜的释放浓度不断发生变化,表明富里酸与铜离子形成的络合物影响磷酸与铜的相互作用,从而影响磷酸盐对铜管的缓蚀效果.     

Isolation and characterization of mesophilic,oxalate-degrading<Emphasis Type="Italic"> Streptomyces</Emphasis> from plant rhizosphere and forest soils

The present work was aimed at the isolation of additional new pure cultures of oxalate-degrading Streptomyces and its preliminary characterization for further work in the field of oxalate metabolism and taxonomic studies. Mesophilic, oxalate-degrading Streptomyces were enriched and isolated from plant rhizosphere and forest soil samples. Strains were examined for cultural, morphological (spore chain morphology, spore mass colour, diffusible and melanin pigment production), physiological (antibiosis, growth in the presence of inhibitory compounds, assimilation of organic acids and enzyme substrates) and chemotaxonomic characters (cellular lipid components and diagnostic cell-wall diamino acid). The taxonomic data obtained were analysed by using the simple matching (S_SM) and Jaccard (S_J) coefficients, clustering was achieved using the UPGMA algorithm. All strains were able to utilize sodium-, potassium-, calcium- and ammonium-oxalate salts. Based on the results of numerical taxonomy, isolates were grouped into five cluster groups with a 70% S_SM similarity level. Streptomyces rochei was the most common of the cluster groups, with a Willcox probability of P>0.8. Streptomyces antibioticus, S. anulatus, S. fulvissimus, S. halstedii and S. violaceusniger are newly reported as oxalate-utilizing Streptomyces.Electronic Supplementary Material  Supplementary material is available for this article if you access the article at . A link in the frame on the left on that page takes you directly to the supplementary material.