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1.
Effects of Cinnamic Acid on Bacterial Community Diversity in Rhizosphere Soil of Cucumber Seedlings Under Salt Stress 总被引:2,自引:0,他引:2
To investigate the effects of a plant autotoxin, cinnamic acid, on bacterial communities in the rhizosphere soil of cucumber seedlings under salt stress, we used cucumber as the experimental material, cinnamic acid as the autotoxin, and NaCl to apply salt stress. Bacterial communities in the rhizosphere soil were analyzed using polymerase chain reaction (PCR), denaturing gradient gel electrophoresis (DGGE), and clone sequencing. Salt stress decreased the diversity of bacterial species in rhizosphere soil of cucumber seedlings at several growth stages. Cinnamic acid exacerbated the effects of salt stress at high concentrations, but alleviated its effects at low concentrations. Cloning and sequencing results indicated that DGGE bands amplified from soil samples showed high homology to uncultured bacterial species. Cinnamic acid at 50 mg kg^-1 soil improved cucumber growth and was the most effective treatment to alleviate the effects of salt stress on bacterial communities. 相似文献
2.
利用免培养的16S rDNA梯度凝胶电泳技术(DGGE)研究脱脂蚕蛹替代鱼粉对建鲤(Cyprinus carpio var. jian)肠道菌群的影响。6个饲养组分别饲喂6种等氮等能饲料, 分别为含10%鱼粉的基础饲料、3种替代饲料(脱脂蚕蛹替代鱼粉蛋白, 替代水平分别为25%、50%和75%)及2组分别在50%和75%替代基础上分别再添加0.6%和0.7%晶体赖氨酸的饲料 (分别记为DSP0、DSP25、DSP50、DSP50-Lys、DSP75、DSP75-Lys)。饲养结束后采集肠道样品, 分析建立指纹图谱, DSP0、DSP25、DSP50、DSP50-Lys、DSP75、DSP75-Lys组分别产生了19、15、11、12、8和12条可鉴别条带, 各组均存在优势种群条带。DSP0和其他各组(DSP25、DSP50、DSP50-Lys、DSP75、DSP75-Lys)的相似性指数为50.7%、50.6%、69.0%、42.4%、66.7%。对PCR-DGGE指纹图谱主要条带进一步回收、克隆和测序, 共得到19条序列, 将得到的序列在NCBI数据库中进行同源分析, 发现建鲤肠道菌群归为芽孢杆菌纲、螺旋体纲、β-变形菌纲、梭菌纲、放线菌纲、γ-变形菌纲、蓝藻纲、拟杆菌纲、梭杆菌纲等, 大部分为不可培养细菌。研究发现, 脱脂蚕蛹替代鱼粉对建鲤肠道菌群组成产生显著影响, 随着脱脂蚕蛹替代水平的提高, 肠道菌群多样性下降, 细菌种类从19种锐减至8种, 而饲料中补充赖氨酸可减弱这种影响, 本研究可为蚕蛹在鲤配合饲料中的开发应用提供参考资料。
3.
Wenting Zhou Xiaoning Nan Zhou Zheng Cong Wei Hong He 《Journal of insect science (Online)》2015,15(1)
Intestinal bacterial community plays a crucial role in the nutrition, development, survival, and reproduction of insects. When compared with other insects with piercing-sucking mouthparts, the habitats of cicada nymphs and adults are totally different. However, little is known about the differences in the gut bacterial communities in the nymphs and adults within any cicada species. The diversity of bacteria in the gut of nymphs and adults of both genders of Meimuna mongolica (Distant) was studied using the denaturing gradient gel electrophoresis (DGGE) method. Few inter-individual variations among gut microbiota were observed, suggesting that M. mongolica typically harbors a limited and consistent suite of bacterial species. Bacteria in the genera Pseudomonas and Enterobacter were the predominant components of the gut microflora of M. mongolica at all life stages. Bacteria of Pantoea, Streptococcus, and Uruburuella were also widespread in the cicada samples but at relatively lower concentrations. The relative stability and similarity of the PCR-DGGE patterns indicate that all individuals of this cicada species harbor a characteristic bacterial community which is independent from developmental stages and genders. Related endosymbionts that could be harbored in bacteromes of cicadas were not detected in any gut samples, which could be related to the cicada species and the distribution of these endosymbionts in the cicada cavity, or due to some of the possible limitations of PCR-DGGE community profiling. It is worthwhile to further address if related cicada endosymbiont clades distribute in the alimentary canals and other internal organs through diagnostic PCR using group-specific primer sets. 相似文献
4.
本研究采用变性梯度凝胶电泳(PCR-DGGE)技术研究猪场沼气池细菌群落,对基于16S rDNAV3区的PCR-DGGE电泳的最佳变性剂梯度范围、电泳时间、染色时间进行优化。研究结果表明,最佳变性剂梯度范围为35%~60%,电泳时间为12h,SYBR Green Fluorescent Dye染料的染色时间是30min,优化后的PCR-DGGE确保了实验的准确性、灵敏度和可重复性。运用此优化后的PCR-DGGE技术对5个猪场沼液细菌群落进行了研究,获得了较丰富的多样性。该实验为深入研究畜禽养殖粪污治理的菌种筛选奠定了基础。 相似文献
5.
Apple replant disease (ARD) is a disease complex that reduces survival, growth and yield of replanted trees, and is often encountered in establishing new orchards on old sites. Methyl bromide (MB) has been the fumigant used most widely to control ARD, but alternatives to MB and cultural methods of control are needed. In this experiment, we evaluated the response of soil microbial communities and tree growth and yield to three pre-plant soil treatments (compost amendment, soil treatment with a broad-spectrum fumigant, and untreated controls), and use of five clonal rootstock genotypes (M.7, M.26, CG.6210, G.30 and G.16), in an apple replant site in Ithaca, New York. Polymerase chain reaction (PCR)—denaturing gradient gel electrophoresis (DGGE) analysis was used to assess changes in the community composition of bacteria and fungi in the bulk soil 8, 10, 18 and 22 months after trees were replanted. PCR-DGGE was also used to compare the community composition of bacteria, fungi and pseudomonads in untreated rhizosphere soil of the five rootstock genotypes 31 months after planting. Tree caliper and extension growth were measured annually in November from 2002 to 2004. Apple yield data were recorded in 2004, the first fruiting year after planting. Trees on CG.6210 rootstocks had the most growth and highest yield, while trees on M.26 rootstocks had the least growth and lowest yield. Tree growth and yield were not affected by pre-plant soil treatment except for lateral extension growth, which was longer in trees growing in compost-treated soil in 2003 as compared to those in the fumigation treatment. Bulk soil bacterial PCR-DGGE fingerprints differed strongly among the different soil treatments 1 year after their application, with the fingerprints derived from each pre-plant soil treatment clustering separately in a hierarchical cluster analysis. However, the differences in bacterial communities between the soil treatments diminished during the second year after planting. Soil fungal communities converged more rapidly than bacterial communities, with no discernable pattern related to pre-plant soil treatments 10 months after replanting. Changes in bulk soil bacterial and fungal communities in response to soil treatments had no obvious correlation with tree performance. On the other hand, rootstock genotypes modified their rhizosphere environments which differed significantly in their bacterial, pseudomonad, fungal and oomycete communities. Cluster analysis of PCR-DGGE fingerprints of fungal and pseudomonad rhizosphere community DNA revealed two distinct clusters. For both analyses, soil sampled from the rhizosphere of the two higher yielding rootstock genotypes clustered together, while the lower yielding rootstock genotypes also clustered together. These results suggest that the fungal and pseudomonad communities that have developed in the rhizosphere of the different rootstock genotypes may be one factor influencing tree growth and yield at this apple replant site. 相似文献
6.
种植水稻和长期施用无机肥对红壤氨氧化细菌多样性和硝化作用的影响 总被引:6,自引:0,他引:6
以中国科学院红壤生态试验站发育于第四纪红黏土的植稻红壤为研究对象,研究了长期种植水稻和施用无机肥对土壤β-变形杆菌纲中氨氧化细菌多样性和硝化作用的影响。原始红壤改种水稻13年后,氨氧化细菌16SrDNA的DGGE条带数量增加,条带谱与原始红壤的差异较大,相似性为61%,说明种植水稻后土壤氨氧化细菌群落结构发生了变化。PCR-DGGE方法研究结果也显示,长期施用无机氮肥的处理(NP、NPK和NK),DGGE带谱相似性较高,达到73%,硝化率和硝化势均高于未施用氮肥的处理。逐步回归分析显示硝化率和硝化势均随着土壤脲酶活性的提高而显著增加。推测尿素可提高土壤水解氮含量,使土壤脲酶活性提高,促进硝化细菌的生长,进而提高硝化率和硝化势。 相似文献
7.
[目的]优选人参根际土壤微生物的PCR-DGGE反应体系。[方法]以种植1年的人参土壤为材料,利用PCR-DGGE技术,对反应体系中的几种重要参数不同梯度进行了优化研究,包括模板、dNTPs、引物及Mg2+的用量。[结果]最适宜的反应体系为:模板DNA浓度为0.8μg/μl,dNTPs浓度为0.2 mmol/L,引物浓度为0.4μmol/L,Mg2+浓度为1.5 mmol/L。[结论]该方法简便快捷,为进一步研究人参根际土壤微生物多样性奠定了基础。 相似文献
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【目的】分析柑橘木虱(Diaphorina citri)体内可能与黄龙病菌(Candidatus Liberibacter asiaticus)互作的内生细菌,为黄龙病菌的人工培养及其病害防控奠定基础。【方法】首先通过传统分离培养方法比较不同地理来源带黄龙病菌(带菌)和不带菌黄龙病菌(不带菌)的木虱中可培养内生细菌的差异。其次将带菌状况不同的木虱分别分为头、胸、腹3部分,经PCR扩增其16S rDNA的V6-V8区,用变性梯度凝胶电泳(PCR-DGGE)方法,比较带菌状况不同的木虱内生细菌的差异和木虱不同部位内生细菌的差异。选择3种差异的内生细菌:Bacillus sp.、Salmonella sp.、Enterobacter sp.,在8份带菌状况不同的木虱样品中通过q-PCR分别对其进行实时荧光定量分析,再以总细菌量为校正计算包括黄龙病菌在内的4种细菌的相对含量,数据经LSD检验,以各种细菌相对含量的-lg值作图,先比较同一样品中3种细菌分别与黄龙病菌的相对含量关系,再比较同种细菌在不同样品中的特性,分析3种内生细菌和黄龙病菌的互作关系。【结果】不带菌木虱中可培养内生菌菌落丰富度和菌落形成单位均大于带菌木虱中。在不带菌木虱中共获得14株形态不同的菌株,分属于芽孢杆菌属(Bacillus,3株)、欧文氏菌属(Erwinia,1株)、克雷伯氏杆菌属(Klebsiella,1株)、葡萄球菌属(Staphylococcus,2株)、节杆菌属(Arthrobacter,1株)、泛菌属(Pantoea,2株)、果胶杆菌属(Pectobacterium,1株)、沙门氏菌属等(Salmonella,1株)、链霉菌属(Streptomyces,1株)、Massilia brevitalea(1株)等10个细菌属。在带菌木虱中分得的4株细菌在不带菌木虱中均分离到,分属于克雷伯氏菌属、芽孢杆菌属、果胶杆菌属。其中Dc-11(嗜气芽孢杆菌属)在带菌木虱和不带菌木虱中分离频率均达到100%,表明其为木虱体内常驻细菌;对木虱不同部位(头、胸、腹)内生细菌16S rDNA的PCR-DGGE图谱显示,带菌与不带菌木虱中细菌种群差别明显,带菌状况相同的木虱不同部位之间差别不明显。其中优势条带10 (Wolbachia sp.)、12(Wolbachia pipientis)、13(Syncytium endosymbiont of Diaphorina citri)、14(Uncultured bacterium)、19(Serratia marcescens)、21和22(均为嗜麦芽寡养单胞菌Stenotrophomonas maltophilia)在木虱体内稳定存在,带菌木虱腹部特有优势内生细菌为Enterobacter sp.,木虱中同样也存在次级内生菌Wolbachia。q-PCR的结果验证了所选的3种细菌在前期传统分离培养和16S rDNA-PCR-DGGE中结果的可靠性,同时表明肠杆菌属在8份样品中与黄龙病菌呈正相关关系。【结论】黄龙病菌进入木虱体内会改变木虱内生细菌菌群种类和结构;嗜气芽孢杆菌(B. aerophilus)在柑橘木虱体内稳定存在,为木虱体内常驻菌群;Enterobacter sp.与黄龙病菌带菌量呈正相关,推测其可能与黄龙病菌互作。 相似文献
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[目的]富集厌氧除磷菌,并对功能菌的组成进行研究。[方法]利用厌氧连续流反应器,以养殖场新鲜鸡粪为种泥,控制和保持反应器内ORP为-337~-230 mV,pH 6~7,温度30~35℃,富集厌氧除磷功能菌。跟踪监测反应器进出水的总磷(TP)、磷酸盐(PO43--P)和氨氮(NH2-N)去除率。当TP的去除率达到60.89%时,取样进行PCR-DGGE分析,用MEGA 4.0软件构建系统进化树。[结果]鸡粪在反应器中连续培养140 d后,TP的去除率平均达到39.86%,大大高于文献报道的24.19%,PO43--P去除率平均达到40.82%,NH3-N的去除率平均达到34.39%;NH3-N与TP的去除率之间存在一定的相关性,厌氧条件下可达到同时脱氮(去除氨氮)除磷(还原磷酸生成磷化氢)的效果;通过对样品的PCR-DGGE分析和进化树分析,获得一种具发酵功能的乳球菌属(Lactoccus)和一种具有发酵、固氮和厌氧除磷功能的梭菌属(Clostridium)。[结论]该研究可为厌氧除磷机理提供理论依据。 相似文献
10.
以餐厨垃圾、果蔬垃圾、麦秸3种不同原料分别进行厌氧消化,研究了各反应器在最佳运行条件下的消化特性和微生物群落组成。结果表明:VS产气率由高到低依次为餐厨垃圾(756.4mL·g-1VS-1)、麦秸(696.5mL·g-1VS-1)和果蔬垃圾(433.5mL·g-1VS-1),甲烷含量在51.5%~55.1%之间,利用PCR-DGGE技术系统地分析了不同原料消化系统内细菌和古菌的群落结构构成及差异。结果表明,虽然3组样品中细菌和古菌的群落存在相同的优势微生物,但其数量和群落结构差异也较为明显,细菌中以拟杆菌(Bacteroidetes)以及古菌中甲烷鬃菌属(Methanosaeta)和甲烷螺菌属(Methanospirillum)均为样品共有的优势微生物。 相似文献