The Application of Succession Analysis in Combination with EHF Irradiation to the Selective Isolation of Actinomycetes from Soil

A new method employing succession analysis and extremely high frequency (EHF) irradiation is proposed for the selective isolation of actinomycetes from soil. Total actinomycetes were efficiently isolated from soil suspensions irradiated in the wavelength band 4.6–5.8 mm on the 14th and 45th days of succession initiated by soil wetting and from soil suspensions irradiated in the wavelength band 8–11.5 mm on the 7th day of succession. The rare actinomycete genera Actinomadura, Micromonospora, Nonomuraea, Microbispora, Amycolatopsis, Pseudonocardia, Saccharothrix, Streptosporangium, Actinosynnema, Nocardioides, and Saccharopolyspora were isolated by either of the two approaches (succession analysis and EHF irradiation); however, the range of isolated rare actinomycetes was considerably wider when a combination of the two approaches was used. For instance, actinomycetes of the rare genera Actinocorallia, Promicromonospora, Actinoplanes, and Kibdelosporangium were isolated only when EHF irradiation was employed at the early stages of succession.     

高寒地区不同退化草地植被特性和土壤固氮菌群特性及其相关性

选取东祁连山不同退化程度的高寒草地为研究对象,调查研究其植物种类、植被盖度、高度、地上生物量等植物指标以及土壤好气性自生固氮菌和嫌气性自生固氮菌数量,在此基础上,采用real-time PCR的方法扩增nifH基因,测定不同退化程度草地土壤中固氮菌相对于土壤总细菌的量,以探讨草地退化过程中植被及土壤固氮菌群的变化规律,结果发现:随着退化程度的加深,草地植物种类逐渐减少,并且优势植物发生变化,毒杂草逐渐增多,植被的高度、盖度、地上生物量都逐渐降低。对土壤固氮菌的研究则表明,土壤好气性自生固氮菌和嫌气性自生固氮菌的数量在不同退化草地随草地退化程度的加重而减少,在同一退化程度草地土壤则是随土层深度加深而下降。对土壤固氮菌nifH基因扩增的结果也表明随着退化加剧,土壤固氮菌相对于土壤总细菌的比例在降低,进一步说明草地退化过程中土壤固氮菌不仅是数量上的下降,更是群落结构层面的变化。对植被特性和土壤固氮菌含量的相关分析表明,植被特性和土壤中固氮菌含量呈显著相关。研究从土壤固氮菌群的角度研究了草地退化的过程,说明了二者具有协同性,研究和治理草地退化必须重视土壤功能菌群尤其是固氮菌群的作用。     

亚热带树种转换对林地土壤微生物群落结构和功能的影响

通过分析杉木采伐迹地营造阔叶树种尾巨桉和固氮树种黑木相思人工林后土壤微生物群落组成和酶活性,探讨造林树种转换对于改善杉木林地土壤微生物特性的影响.结果表明: 树种转换对土壤微生物群落组成和酶活性的影响主要局限于0～10 cm土壤层.杉木转换为固氮树种黑木相思后,显著提高了0～10 cm土壤层总脂肪酸含量、真菌、革兰氏阳性细菌、革兰氏阴性细菌和放线菌生物量.主成分分析表明,黑木相思人工林土壤微生物群落组成与杉木和尾巨桉人工林具有显著差异,土壤中革兰氏阳性细菌、阴性细菌和放线菌丰度显著提高.在0～10 cm土壤层,黑木相思人工林土壤纤维素水解酶、乙酰氨基-葡萄糖苷酶和酸性磷酸酶活性均显著高于杉木和尾巨桉人工林.研究表明,杉木转变为固氮树种黑木相思后会显著提高微生物生物量和酶活性,有助于土壤有机质的恢复,加快养分循环过程.     

Isolation of Actinomycetes from Soil Using Extremely High Frequency Radiation

A new method employing extremely high frequencies (EHFs) is proposed for the selective isolation of actinomycetes from soil. The pretreatment of soil suspensions with EHF wavelengths of 5.6 and 7.1 mm led to a nonselective isolation of actinomycetes. At the same time, the irradiation of soil suspensions within wavelength bands of 3.8–5.8 and 8–11.5 mm considerably augmented the total number of isolated actinomycetes and increased the fraction of the isolated rare genera by 2 and 7 times, respectively. The rare actinomycete genera were represented by Actinomadura, Microtetraspora, Nonomuraea, Micromonospora, Amycolatopsis, Pseudonocardia, Saccharotrix, and Streptosporangium.     

微波处理对嗜碱和嗜盐海洋放线菌分离效果的影响

【目的】研究微波处理对于分离嗜碱和嗜盐海洋放线菌的效果。【方法】用微波处理7份海泥样品,梯度稀释后涂布于3种分离培养基,分离具有嗜碱和嗜盐特性的海洋放线菌。【结果】微波处理后的7份样品中,4份样品中嗜碱海洋稀有放线菌和3份样品的嗜盐海洋稀有放线菌数量极显著提高;7份样品中的嗜碱、嗜盐海洋小单孢菌属、游动放线菌属、诺卡氏菌属等稀有放线菌数量均有显著增加,不同样品中新分离到链孢菌属、小双孢菌属、链孢囊菌属及其他未鉴定的海洋稀有放线菌,分离到属的数量提高了1-4个。【结论】微波处理不仅显著提高嗜碱和嗜盐海洋放线菌的分离数量,而且明显增加了海洋稀有放线菌的分离种类。     

微波处理对土壤放线菌分离效果的影响

采用稀释平皿分离测数及琼脂块法研究了微波预处理对土壤放线菌分离效果的影响.结果表明: 1)微波处理能显著增加可培养放线菌数量.随着微波处理时间的增加,高有机质土壤放线菌总数呈先增后减的趋势.处理3～15 min,GA及HA培养基上放线菌总数分别较对照提高8.3%～92.6%及24.4%～108.5%;处理18～24 min,放线菌总数分别较对照降低了62.1%～78.8%及41.4%～79.8%.微波处理对低有机质土壤放线菌数量无明显影响.2)微波处理对可培养放线菌种类有明显影响.随着微波处理时间的增加,高有机质土壤中新出现的放线菌种类呈先增后减趋势. 处理3～24 min,GA与HA培养基上新出现放线菌种类占放线菌总种类的比例为62.5%～85.7%与66.7%～83.3%,新出现了原小单孢菌属、链轮丝菌属等稀有放线菌属;链霉菌类群也有明显变化.低有机质土壤具有类似趋势.3)微波处理对可培养放线菌中拮抗性放线菌株数占供试放线菌总株数的比例也有明显影响.微波处理6、9和15 min,拮抗性放线菌株数所占比例分别较对照提高66.7%、66.7%和83.3%,其中新出现的拮抗性放线菌株数分别占拮抗性放线菌总株数的70.0%, 90.0%和81.8%.     

南亚热带红椎和格木人工幼龄林土壤微生物群落结构特征

采用氯仿熏蒸浸提法和磷脂脂肪酸法(Phospholipids fatty acid,PLFA)研究了我国南亚热带地区非固氮树种红椎(Castanopsis hystrix)和固氮树种格木(Erythrophleum fordii)人工幼龄林土壤微生物生物量与微生物群落结构特征。结果表明,在旱季和雨季,红椎幼龄林土壤微生物总PLFAs量,细菌PLFAs量、放线菌PLFAs量及丛枝菌根真菌PLFAs量均大于格木幼龄林。红椎幼龄林土壤PLFA Shannon多样性指数(H_(PLFA))在旱季和雨季均大于格木幼龄林。主成分分析表明,土壤微生物群落结构组成受到林分类型和季节的双重影响。冗余分析表明,土壤硝态氮(NO_3~--N)含量、土壤含水量、p H及土壤微生物生物量氮(MBN)与特征磷脂脂肪酸之间呈显著相关关系。以上结果表明固氮树种格木与非固氮树种红椎人工幼龄林对土壤微生物生物量和群落结构的影响存在显著差异。     

The application of succession analysis in combination with extremely high-frequency irradiation to the selective isolation of actinomycetes from soil

A new method employing succession analysis and extremely high-frequency (EHF) irradiation is proposed for the selective isolation of actinomycetes from soil. Total actinomycetes were efficiently isolated from soil suspensions irradiated in the wavelength band 4.6-5.8 mm on the 14th and 45th days of succession initiated by soil wetting and from soil suspensions irradiated in the wavelength band 8-11.5 mm on the 7th day of succession. The rare actinomycete genera Actinomadura, Micromonospora, Nonomuraea, Microbispora, Amycolatopsis, Pseudonocardia, Saccharothrix, Streptosporangium, Actinosynnema, Nocardioides, and Saccharopolyspora were isolated by either of the two approaches (succession analysis and EHF irradiation); however, the range of isolated rare actinomycetes was considerably wider when the combination of the two approaches was used. For instance, actinomycetes of the rare genera Actinocorallia, Promicromonospora, Actinoplanes, and Kibdelosporangium were isolated only when EHF irradiation was employed at the early stages of succession.     

SOME EFFECTS OF GAMMA IRRADIATION ON SEEDS AND RHIZOMES OF MUSA

The tolerance of Musa balbisiana Colla seeds to gamma irradiation was found to be considerably greater than that of rhizomes of the parthenocarpic variety ‘Gros Michel': e.g., 11.8 krad reduced the germination of rhizomes 92% and of seeds approximately 15%. Intact seeds exposed to doses higher than 48 krad did not germinate in non-sterile soil, but, when scarified and cultured axenically after irradiation, seeds which received doses as high as 70 krad germinated. Embryos excised from seeds exposed to doses as high as 285 krad formed callus, indicating that not all metabolic processes were inhibited by these extremely high doses. There was considerable variation in radiation tolerance between seed lots which was not related to their age, moisture content, or pre-exposure viability. Germination of intact seeds appeared to be stimulated by doses of 3 or 9 krad. No lasting differences attributable to the level of irradiation were apparent in the development of seedlings derived from either intact or scarified seeds nor of plantlets derived from excised embryos. Conversely, there was a significant reduction, proportional to irradiation dose, in the growth of plants developing from rhizomes, emphasizing the greater radiation sensitivity of the vegetative propagule. The radiation tolerance of seed-borne microorganisms was considerably higher than that of the plant materials, indicating that gamma irradiation is not effective as a means of obtaining pathogen-free rhizomes or surface-sterilizing seeds of M. balbisiana.     

Microwave irradiation is a useful tool for improving isolation of actinomycetes from soil

Actinomycetes are an important source of novel, biologically active compounds. New methods need to be developed for isolating previously unknown actinomycetes from soil. The objective of this experiment was to study microwave irradiation of soil as a means for isolating previously unknown actinomycetes. Soil samples were collected at ten elevations between 800 m and 3670 m on Taibai Mountain, Shaanxi Province, China. Moistened soil samples were irradiated at 120 W heating power (2450 MHz) for 3 min using a household microwave oven. Irradiation increased total actinomycete, streptomycete, and antagonistic actinomycete counts on three types of culture media. Irradiation also increased the number of culturable actinomycete isolates. Some actinomycete isolates were culturable only after the soil was irradiated, whereas other isolates could not be cultured after irradiation. Irradiation of soil from elevations >3000 m increased actinomycete counts significantly but had little effect on the number of culturable actinomycete isolates. In contrast, irradiation of samples from elevations <3000 m had relatively little effect on actinomycete counts, but significantly increased the number of culturable actinomycete isolates. We used 16S rDNA sequence analysis to identity 14 actinomycete isolates that were only culturable after irradiation. Microwave irradiation of soil was helpful for isolating Streptomyces spp., Nocardia spp., Streptosporangium spp., and Lentzea spp. Slightly more than 90% of the identified actinomycete species were biologically active. In conclusion, microwave irradiation is a useful tool for isolating biologically active actinomycetes from soil.     

Study on the communities of endophytic fungi and endophytic actinomycetes from rice and their antipathogenic activities in vitro

The populations of endophytic fungi and actinomycetes from four rice cultivars in the Panyu district (Site 1) and Wushan district (Site 2) in Guangdong province, South China, were studied. The preponderant endophytic fungi and actinomycetes isolated belonged to Fusarium and Streptomyces respectively. The incidence of Streptomycetes griseofuscus ranged from 36.1 to 69% out of all the different rice cultivars from the two sites. It is the commonest population of endophytic actinomycetes, and constituted the greatest part of all the antagonistic communities. The distributions of endophytic fungi and actinomycetes in roots and leaves were different, endophytic fungi from leaves were diverse, some were organ-specific. More diverse endophytic actinomycetes were isolated from roots than from leaves. The endophytic fungi isolated from rice in Site 2 were more diverse than that in Site 1. The diversity of the endophytic actinomycetes, however, was less than that in Site 1. Acid soil in Site 2 is ideal for the growth and colonization of fungi while the alkaline soil in Site 1 is better for the growth and colonization of actinomycetes. The results suggested that differences in the chemical composition of soil could influence the endophytic microbial communities of rice plants. The endophytic fungi and actinomycetes isolated from poor-growing seedlings and susceptible rice cultivars were more abundant than that the disease-resistant counterparts. In the dual culture and activity detection of the metabolites, 41.2% of all the isolated endophytic fungi showed antagonism to rice pathogens. Fifty percent of all the isolated endophytic actinomycetes were antagonistic to those pathogens. The percentage of Streptomyces griseofuscus and hygroscopicus reached 55.4 and 21.4% of all the active actinomycetes. This revised version was published online in August 2006 with corrections to the Cover Date.     

Mutational approach for N2-fixing and P-solubilizing mutant strains of Klebsiella pneumoniae RSN19 by microwave mutagenesis

Nitrogen (N) fixing Klebsiella pneumoniae RSN19 has high inorganic phosphorus (P) solubilizing capability, but its N₂-fixing capability is limited. In order to acquire a P-solubilizing mutant strain with high efficiency N-fixing capability, different microwave irradiation intensities and durations were tested on RSN19 in an attempt to produce mutants with improved N₂-fixation and P-solubilization capabilities. The effect of microwave irradiation power and time were studied and the microwave mutagenesis parameters were optimized. Nitrogenase activity was tested on the mutant strains by acetylene reduction method; and their P-solubilizing capability and genetic stability were determined. The results indicated that the best conditions for microwave mutagenesis that produced better performed mutant strains were 250W, 36 s. Under these conditions a maximum positive mutation rate of 1.66% was obtained, resulting in five genetically stable strains with promoted nitrogenase activity which was designated as RSM-219, RSM-206, RSM-224, RSM-225 and RSM-275. Subculture tests showed that RSM-219 and RSM-206 were genetically stable mutant strains with higher nitrogenase activity and phosphate solubilizing capabilities than the original strain. Both RSM-219 and RSM-206 performed better than the original strain under N-free conditions when supplied with calcium phosphate only, and produced greater increases in the biomass of alfalfa seedlings.     

Effect of post-irradiation ageing on onion seeds

Seeds of onion (Allium cepa) cv. Ailsa Craig were exposed to various doses of gamma radiation (0, 10, 20, 40, 80 and 100 krad) and subjected to accelerated ageing (RH 100%, 42°C) for 12 h. Radiation doses significantly affected the percentage of normal seedlings, abnormality types (%) and seedling growth. Seed viability, conductivities of seed leachates, final germination (%), germination speed and time to reach 50% germination (T ₅₀) were not affected by the seed irradiation. Accelerated ageing after irradiation had significant influence on seed viability, conductivities of seed leachates, final germination (%) and percentage of normal seedlings. Germination speed, T ₅₀ and seedling growth were not affected by the accelerated ageing. However, all the parameters studied were significantly influenced by the interaction of radiation doses and accelerated ageing. Accelerated ageing generally enhanced the damaging effects of irradiation on seeds. Therefore, it was concluded that onion seeds should not be exposed to adverse environmental conditions after irradiation.     

西沙海藻共附生放线菌的分离鉴定及其抗菌活性评价

【目的】为了探究南海海藻共附生放线菌资源的多样性及潜在的应用价值,对中国西沙群岛来源的海藻进行共附生放线菌的分离鉴定与抗菌活性筛选。【方法】利用稀释涂布平板法,采用2种不同分离培养基对不同采样位点的6种海藻进行放线菌分离;通过16S rRNA基因序列分析、构建系统发育树对分离的放线菌进行鉴定;用打孔法对无乳链球菌(Streptococcus agalactiae)等10种敏感细菌进行抗菌活性筛选;对筛选得到的目标活性菌株HZ014进行全基因组测序,通过AntiSMASH在线工具分析其次级代谢产物生物合成基因簇,预测其产生新型活性物质的潜力。【结果】从6种海藻中分离得到36株共附生放线菌,基于16S rRNA基因序列比对和系统发育分析,鉴定结果为链霉菌属(Streptomyces) 2株、红球菌属(Rhodococcus) 2株、诺卡氏菌属(Nocardia)3株、小单孢菌属(Micromonospora) 5株和盐孢菌属(Salinispora) 24株;抗菌活性筛选结果表明,36株共附生放线菌发酵粗提物对至少1种敏感细菌表现出一定的抑制作用,不同菌株发酵粗提物的抗菌活性存在明显差异,...     

The effect of gamma irradiation on potato microtuber production in vitro

The effects of low doses of gamma irradiation and potato (Solanum tuberosum L.) cultivar on the production of microtubers in vitro were investigated. Nodal segments from virus free explants of three potato cultivars (cv.) were placed on tuberization inducing medium and irradiated with 4 doses of gamma radiation (2.5, 5, 10, 15 Gy). Cv. Diamant produced the highest number of microtubers followed by Draga and Spunta. Irradiation of the explants with 2.5 Gy of gamma radiation led to a significant increase in the number of microtubers (38% increase over the control). Average weight of microtubers was not significantly influenced by low doses of gamma irradiation. Draga microtubers were the largest followed by Diamant and Spunta. Microtubers resembled mature tubers in shape (Spunta was oval and Draga and Diamant were spherical). Size of microtubers was crucial for sprouting in vivo. It is suggested that only microtubers larger than 5 mm in diameter (250 mg) be used to produce minitubers in vivo. Since 2.5 Gy is a low irradiation dose, it can be used to enhance tuberization in vitro without fear of genetic changes in the used cultivars. This revised version was published online in June 2006 with corrections to the Cover Date.     

Detection of Eubacterial 3-Hydroxy-3-Methylglutaryl Coenzyme A Reductases from Natural Populations of Actinomycetes

Three natural populations of actinomycetes were investigated by PCR for the presence of type I 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG CoA), a gene associated with isoprenoid biosynthesis. The populations were obtained from an agricultural site (69 isolates), a coastal salt marsh (220 isolates), and a desert soil (96 isolates). A set (34) of standard actinomycete reference strains were also investigated. The target gene was only detected in 5 of the 419 actinomycetes screened, which represented 4 from the coastal salt marsh and one reference strain. The isolates that contained the gene were taxonomically diverse (4 Streptomyces spp. and 1 Nocardia sp.). These results suggest that type I HMG CoA containing pathways are rare in actinomycetes and their distribution within actinomycetes populations is not random.     

乔木根系和凋落物对南亚热带3种人工林土壤线虫群落的差异化影响

根系分泌物和凋落物为土壤食物网提供了基础的养分资源。然而,不同树种纯林和混交林地下根系和地上凋落物对土壤线虫群落的影响机制尚不清楚。2019年9月在广西凭祥热带林业实验中心选取格木(Erythrophleum fordii Oliv.)纯林、马尾松(Pinus massoniana Lamb.)纯林和格木×马尾松混交林3种林分类型,分别设置对照、阻断乔木根系、去除地上凋落物和阻断乔木根系并去除地上凋落物四组实验处理,于2021年3月对3种林分类型不同处理下的土壤线虫群落和土壤理化性质进行了调查。研究结果表明,无论哪种林分类型,阻断根系改变了土壤线虫群落的营养类群组成,显著降低了食真菌线虫相对多度,增加了植物寄生线虫相对多度;去除凋落物显著降低了土壤线虫密度、类群数、线虫通路比值和结构指数,增加了基础指数,表明去除凋落物降低了土壤食物网的稳定性。无论哪种林分类型,人工林中树木地下根系输入是构建土壤线虫群落营养类群组成的主要驱动因素,地上凋落物在维持土壤食物网稳定性方面发挥着重要的作用。此外,阻断根系和去除凋落物对混交林中土壤线虫群落没有显著的影响,表明含固氮树种的格木×马尾松混交林比人工...     

DNA degradation and reduced recombination following UV irradiation during meiosis in yeast (Saccharomyces cerevisiae)

Summary Irradiation of meiotic yeast cells with moderate doses of ultraviolet irradiation (1,600 erg/mm²) leads to the arrest of premeiotic DNA synthesis, massive (5–40%) DNA degradation, and a 40–50% loss of cell viability. In contrast, such doses of UV irradiation had a minor effect on viability (15–20% loss) of logarithmically growing cells, and no comparable DNA degradation was observed in irradiated synchronized vegetative cells. Meiotic recombination is also affected by UV irradiation. When administered at a stage comparable to meiotic prophase, low doses of irradiation result in a reduction in recombination frequency without significantly affecting cell viability.     

Isolation and characterization of actinomycetes from healthy goat faeces

Aims: To isolate and characterize actinomycetes with probiotic activities from healthy goat faeces. Methods and Results: Faecal actinomycetes were isolated by dilution methods and identified by 16S rRNA gene sequence analysis. The hydrolytic enzyme activities were analysed by clear zone formation. The antimicrobial activities and resistance to heavy metals were tested by growth inhibition methods. The isolates belong to a small group of actinobacterial genera, including Streptomyces, Nocardiopsis and Oerskovia. The Oerskovia was the most widely distributed genus among the cultures. The proportion of streptomycete‐like strains producing amylase or protease is significantly higher than those of other actinomycetes (P < 0·05). Compared with streptomycete‐like strains, a higher proportion of (α‐ or β‐) galactase‐producing other actinomycetes was found in goat faeces. More than 50% of streptomycete‐like strains showed activities against test fungi. Streptomycetes could tolerate 0·25 mmol l^?1 Cr₂O₇^2?, 2 mmol l^?1 Ni²⁺; however, other actinomycetes are liable to 40 mmol l^?1 Fe³⁺ and 0·25 mmol l^?1 Cr₂O₇^2? and resistant to 5 mmol l^?1 Ni²⁺ and 2 mmol l^?1 Cu²⁺. Conclusions: The different physiological characteristics of the actinomycetes suggested that the cooperation in the actinomycetes might be involved in their association with goat. Significance and Impact of the Study: Probiotic mixtures based on faecal actinomycetes showed potentials in animal production.     

Saprophytic Actinomycetes Promote Nodulation in <Emphasis Type="Italic">Medicago sativa</Emphasis>-<Emphasis Type="Italic">Sinorhizobium meliloti</Emphasis> Symbiosis in the Presence of High N

Saprophytic rhizoactinomycetes isolated from the root nodule surface of the nitrogen-fixing actinorhizal plant Discaria trinervis, Streptomyces MM40, Actinoplanes ME3, and Micromonospora MM18, previously shown to stimulate nodulation in Frankia-Discaria trinervis symbiosis, were assayed as co-inoculants with Sinorhizobium meliloti 2011 on Medicago sativa. When plants were fertilized with a low level of N (0.07 mM), the inoculation of the actinomycetes alone did not show any effect on plant growth. Meanwhile, when actinomycetes were co-inoculated with S. meliloti, nodulation and plant growth were significantly stimulated compared to plants inoculated with only S. meliloti. The analysis of nodulation kinetics of simultaneously or delayed co-inoculations suggests that the effect of the actinomycetes operates in early infection and nodule development counteracting the autoregulation of nodulation by the plant. Because the actinomycete effect was found in the symbiotic nitrogen-fixing state of the plant, we investigated the effects of the actinomycetes, in single inoculation or co-inoculation with S. meliloti, on plants grown under a high level of N (7 mM) that was inhibitory for nodulation by S. meliloti. The inoculation of the actinomycetes alone did not show any effect on plant growth although high N was available. Unexpectedly, the co-inoculation of actinomycetes with S. meliloti on plants grown with high N (7 mM) significantly stimulates nodulation, clearly counteracting the inhibition of nodulation by high N. These results corroborate that the interaction of rhizoactinomycetes would interfere with the autoregulation of nodulation in alfalfa mediated by high N, opening new research lines of potential agronomical applications.