Assessing Genotypic Diversity and Symbiotic Efficiency of Five Rhizobial Legume Interactions Under Cadium Stress for Soil Phytoremediation

In the framework of soil phytoremediation using local legume plants coupled with their native root-nodulating bacteria to increase forage yields and preserve contaminated soils in arid regions of Tunisia, we investigated the diversity of bacteria from root nodules of Lathyrus sativus, Lens culinaris, Medicago marina, M. truncatula, and M. minima and the symbiotic efficiency of these five legume symbiosis under Cadmium stress. Fifty bacterial strains were characterized using physiological and biochemical features such heavy metals resistant, and PCR-RFLP of 16S rDNA. Taxonomically, the isolates nodulating L. sativus, and L. culinaris are species within the genera Rhizobium and the ones associated to Medicago sp, within the genera Sinorhizobium. The results revealed also that the cadmium tolerance of the different legumes-rhizobia interaction was as follows: M. minima<M. truncatula<M. marina<L. sativus<L. culinaris indicating that the effect of Cadmium on root nodulation and biomass production is more deleterious on M. minima-S. meliloti and M. truncatula-S. meliloti than in other symbiosis. Knowledge on genetic and functional diversity of M. marina, L. sativus and L. culinaris microsymbiotes is very useful for inoculant strain selection and can be selected to develop inoculants for soil phytoremediation.     

Influence of the Size of Indigenous Rhizobial Populations on Establishment and Symbiotic Performance of Introduced Rhizobia on Field-Grown Legumes

Indigenous rhizobia in soil present a competition barrier to the establishment of inoculant strains, possibly leading to inoculation failure. In this study, we used the natural diversity of rhizobial species and numbers in our fields to define, in quantitative terms, the relationship between indigenous rhizobial populations and inoculation response. Eight standardized inoculation trials were conducted at five well-characterized field sites on the island of Maui, Hawaii. Soil rhizobial populations ranged from 0 to over 3.5 × 10⁴ g of soil^-1 for the different legumes used. At each site, no less than four but as many as seven legume species were planted from among the following: soybean (Glycine max), lima bean (Phaseolus lunatus), cowpea (Vigna unguiculata), bush bean (Phaseolus vulgaris), peanut (Arachis hypogaea), Leucaena leucocephala, tinga pea (Lathyrus tingeatus), alfalfa (Medicago sativa), and clover (Trifolium repens). Each legume was (i) inoculated with an equal mixture of three effective strains of homologous rhizobia, (ii) fertilized at high rates with urea, or (iii) left uninoculated. For soybeans, a nonnodulating isoline was used in all trials as the rhizobia-negative control. Inoculation increased economic yield for 22 of the 29 (76%) legume species-site combinations. While the yield increase was greater than 100 kg ha^-1 in all cases, in only 11 (38%) of the species-site combinations was the increase statistically significant (P ≤ 0.05). On average, inoculation increased yield by 62%. Soybean (G. max) responded to inoculation most frequently, while cowpea (V. unguiculata) failed to respond in all trials. Inoculation responses in the other legumes were site dependent. The response to inoculation and the competitive success of inoculant rhizobia were inversely related to numbers of indigenous rhizobia. As few as 50 rhizobia g of soil^-1 eliminated inoculation response. When fewer than 10 indigenous rhizobia g of soil^-1 were present, economic yield was significantly increased 85% of the time. Yield was significantly increased in only 6% of the observations when numbers of indigenous rhizobia were greater than 10 cells g of soil^-1. A significant response to N application, significant increases in nodule parameters, and greater than 50% nodule occupancy by inoculant rhizobia did not necessarily coincide with significant inoculation responses. No less than a doubling of nodule mass and 66% nodule occupancy by inoculant rhizobia were required to significantly increase the yield of inoculated crops over that of uninoculated crops. However, lack of an inoculation response was common even when inoculum strains occupied the majority of nodules. In these trials, the symbiotic yield of crops was, on average, only 88% of the maximum yield potential, as defined by the fertilizer N treatment. The difference between the yield of N-fertilized crops and that of N₂-fixing crops indicates a potential for improving inoculation technology, the N₂ fixation capacity of rhizobial strains, and the efficiency of symbiosis. In this study, we show that the probability of enhancing yield with existing inoculation technology decreases dramatically with increasing numbers of indigenous rhizobia.     

Symbiotic relationships between ants and aphids

The classical and contemporary literature devoted to symbiotic relationships between ants and aphids was analyzed. The most studied issues are: the totality of morphological and anatomical adaptations of partners to symbiosis, different degrees of aphids myrmecophilia, benefits and costs of insects during interaction. A lot of issues are still discussible. For instance, it is unknown whether the trophobiotical relationships between ants and aphids are mutualistic or intermediate between mutualism and exploitation. To clarify this vague point the study of symbionts behaviour is of great importance, however, it remains practically non-investigated either for aphids or ants. It is known only that nonmyrmecophilous aphids living within galls have rather complicated behaviour--their colonies are guarded with specialized castes of aphids-soldiers. As for the ants, till nowadays trophobionts (individuals looking after the aphids), which make up special functional group within ants family, have been described as passive foragers being occupied only with the honeydew collection and transportation. The recent investigations revealed that ants use behavioural schemes with different degree of complication interacting with aphids: from lonely foraging to professional specialization in working groups.     

Physiological Characteristics of Cowpea Rhizobia: Evaluation of Symbiotic Efficiency in Vigna unguiculata

One fast-growing and three slow-growing strains of Rhizobium (isolated from cowpeas) were evaluated for symbiotic performance on Vigna unguiculata (L.) Walp. cultivar California no. 5 blackeyes. Plants inoculated with slow-growing strains 176A22, 176A30, and 176A32 developed a maximum acetylene reduction activity of 24.6, 27.0, and 32 μmol of ethylene formed per plant per h, respectively, versus 6.4 μmol per plant per h in plants inoculated with the fast-growing strain 176A28. When inoculated with approximately equal proportions of rhizobia, the fast-growing strain 176A28 produced 95% of the nodules when challenged with the slow-growing strain 176A22, but formed only 6% of the nodules when challenged with the slow-growing strain 176A30. Consequently, there was no relation between the growth rate in vitro and the capability of rhizobia to compete for nodule-forming sites. Plants inoculated with strain 176A28 and subjected to drought during the vegetative growth period recovered to the same level of nitrogen fixation and nodulation as those that received adequate irrigation. On the other hand, plants inoculated with strains 176A22, 176A30, and 176A32 failed to achieve the same levels of nodulation and nitrogen fixation under drought as compared with irrigated conditions.     

Symbiotic interactions between soybean and competing strains ofBradyrhizobium japonicum

Selected symbiotic characteristics of fiveBradyrhizobium japonicum strains were assessed in association with ‘Ransom’ soybean plants (Glycine max [L.] Merr.). In the first of two greenhouse experiments, relative nodulation competitiveness of the strains was examined. Strains were grouped into pairs, and corresponding cells were applied to surface-disinfected seeds so as to provide seven ratios of cell numbers between the two strains. Tap root nodules were harvested 28 days after sowing and serotyped by means of an enzyme-linked immunosorbent assay. Strains differed considerably in nodulation competitiveness, and these differences were successfully quantified using relationships previously proposed in the literature. A second experiment involved assessment of the reproducibility of this technique and characterization of the symbiotic response to single- and double-strain inocula. Differences in relative nodulating abilities of strains were apparent between experiments and were possibly related to observed variations in greenhouse temperatures. Plant shoot weight and total N content were not significantly correlated with nodule number or weight when evaluated across inoculation treatments, but these correlations were often significant within inoculation treatments. Certain double-strain inocula produced either positive or negative effects on shoot weight, N content, and nodulation, when compared with values predicted from corresponding controls receiving single-strain inocula. Paper No. 11741 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, NC 27695-7643, USA.     

影响根瘤菌共生固氮效率的主要因素及遗传改造

介绍了影响根瘤菌共生固氮效率的主要因素及遗传改造,包括土壤因素,宿主植物,四碳二羟酸转移酶基因dct,固氮调节基因nifA,吸氢酶基因hup,共生质粒（基因）,缺陷型回复突变等。     

Ontogenetic Interactions between Photosynthesis and Symbiotic Nitrogen Fixation in Legumes

Photosynthetic data collected from Pisum sativum L. and Phaseolus vulgaris L. plants at different stages of development were related to symbiotic N₂ fixation in the root nodules. The net carbon exchange rate of each leaf varied directly with carboxylation efficiency and inversely with the CO₂ compensation point. Net carbon exchange of the lowest leaves reputed to supply fixed carbon to root nodules declined in parallel with H₂ evolution from root nodules. The decrease in H₂ evolution also coincided with the onset of flowering but preceded the peak in N₂ fixation activity measured by acetylene-dependent ethylene production. A result of these changes was that the relative efficiency of N₂ fixation in peas increased to 0.7 from an initial value of 0.4. The data reveal that attempts to identify photosynthetic contributions of leaves to root nodules will require careful timing and suggest that the relative efficiency of N₂ fixation may be influenced by source-sink relationships.     

Phenotypic,Molecular and Symbiotic Characterization of the Rhizobial Symbionts of Desmanthus paspalaceus (Lindm.) Burkart That Grow in the Province of Santa Fe,Argentina

Desmanthus paspalaceus (Lindm.) Burkart belongs to the D. virgatus complex, subfamily Mimosoidae. The known potential as livestock fodder of several of these legumes prompted us to undertake a phenotypic, molecular, and symbiotic characterization of the D. paspalaceus symbionts in the Santa Fe province, Argentina. The rhizobia collected—containing isolates with different abiotic-stress tolerances—showed a remarkable genetic diversity by PCR fingerprinting, with 11 different amplification profiles present among 20 isolates. In selected isolates 16S-rDNA sequencing detected mesorhizobia (60%) and rhizobia (40%) within the collection, in contrast to the genus of the original inoculant strain CB3126—previously isolated from Leucaena leucocephala—that we typified here through its 16S rDNA as Sinorhizobium terangae. The results revealed the establishment by diverse bacterial genera -rhizobia, sinorhizobia, and mesorhizobia- of full N₂-fixing symbiotic associations with D. paspalaceus. This diversity was paralleled by the presence of at least two different nodC allelic variants. The identical nodC alleles of the Mesorhizobia sp. 10.L.4.2 and 10.L.5.3 notably failed to group within any of the currently described rhizo-/brady-/azorhizobial nodC clades. Interestingly, the nodC from S. terangae CB3126 clustered close to homologs from common bean nodulating rhizobia, but not with the nodC from S. terangae WSM1721 that nodulates Acacia. No previous data were available on nod-gene phylogeny for Desmanthus symbionts. A field assay indicated that inoculation of D. paspalaceus with the local Rhizobium sp. 10L.11.4 produced higher aerial-plant dry weights compared to S. teranga CB3126–inoculated plants. Neither the mesorhizobia 10.L.4.2 or 10.L.5.3 nor the rhizobium 10L.11.4 induced root nodules in L. leucocephala or P. vulgaris. The results show that some of the local isolates have remarkable tolerances to several abiotic stresses including acidity, salt, and temperature; while exhibiting prominent N₂ fixation; thus indicating suitability as candidates for inoculation of D. paspalaceus.     

The Symbiotic Anthozoan: A Physiological Chimera between Alga and Animal

The symbiotic life style involves mutual ecological, physiological,structural, and molecular adaptations between the partners.In the symbiotic association between anthozoans and photosyntheticdinoflagellates (Symbiodinium spp., also called zooxanthellae),the presence of the endosymbiont in the animal cells has constrainedthe host in several ways. It adopts behaviors that optimizephotosynthesis of the zooxanthellae. The animal partner hashad to evolve the ability to absorb and concentrate dissolvedinorganic carbon from seawater in order to supply the symbiont'sphotosynthesis. Exposing itself to sunlight to illuminate itssymbionts sufficiently also subjects the host to damaging solarultraviolet radiation. Protection against this is provided bybiochemical sunscreens, including mycosporine-like amino acids,themselves produced by the symbiont and translocated to thehost. Moreover, to protect itself against oxygen produced duringalgal photosynthesis, the cnidarian host has developed certainantioxidant defenses that are unique among animals. Finally,living in nutrient-poor waters, the animal partner has developedseveral mechanisms for nitrogen assimilation and conservationsuch as the ability to absorb inorganic nitrogen, highly unusualfor a metazoan. These facts suggest a parallel evolution ofsymbiotic cnidarians and plants, in which the animal host hasadopted characteristics usually associated with phototrophicorganisms.     

黄花杓兰与菌根真菌共生关系研究

对黄花杓兰(Cypripedlium flavum P.F.Hunt et Summerh.)植株的根进行了一个生长周期的采集和切片观察,发现黄花杓兰具有相应的菌根结构,菌丝的存在状态在各物候期有所不同,可分为5个时期：(1)4月份联通菌丝时期;(2)5月份菌丝团时期Ⅰ;(3)6、7月份无菌丝团时期I;(4)8、9月份菌丝团时期Ⅱ;(5)10月份无菌丝团时期Ⅱ。可以看出,在植株生活转变期,根细胞内出现菌丝团;在植株完成正常的生命过程中,真菌营养是光合营养的补充;真菌菌丝团也作为植物营养的储存器,菌丝团消解释放营养供给植株。在此基础上探讨了菌根真菌与黄花杓兰的共生关系的一个方面：植株通过联通菌丝沟通各个细胞的物质交换,菌根真菌利用植株体内物质完成生命活动,菌丝团消解,使真菌所储藏的营养物转化,为植株所利用。     

Comparing parasite numbers between samples of hosts

The comparison of parasite numbers or intensities between different samples of hosts is a common and important question in most parasitological studies. The main question is whether the values in one sample tend to be higher (or lower) than the values of the other sample. We argue that it is more appropriate to test a null hypothesis about the probability that an individual host from one sample has a higher value than individual hosts from a second sample rather than testing hypotheses about means or medians. We present a recently proposed statistical test especially designed to test hypotheses about that probability. This novel test is more appropriate than other statistical tests, such as Student's t-test, the Mann-Whitney U-test, or a bootstrap test based on Welch's t-statistic, regularly used by parasitologists.     

Efficiency of Glasshouse Pot Experiments Rotating versus Stationary Benches

Pot cultures of rice (Oryza sativa L., var. Colusa) in flooded soil were used to measure comparative uniformity of growth where mutual shading was the primary source of variation. Data were analyzed according to various divisions of each layout into blocks, sections, and replicate pots to compare statistical efficiencies.     

Comparing phylogenetic codivergence between polyomaviruses and their hosts

Seventy-two full genomes corresponding to nine mammalian (67 strains) and two avian (5 strains) polyomavirus species were analyzed using maximum likelihood and Bayesian methods of phylogenetic inference. Our fully resolved and well-supported (bootstrap proportions > 90%; posterior probabilities = 1.0) trees separate the bird polyomaviruses (avian polyomavirus and goose hemorrhagic polyomavirus) from the mammalian polyomaviruses, which supports the idea of spitting the genus into two subgenera. Such a split is also consistent with the different viral life strategies of each group. Simian (simian virus 40, simian agent 12 [Sa12], and lymphotropic polyomavirus) and rodent (hamster polyomavirus, mouse polyomavirus, and murine pneumotropic polyomavirus [MPtV]) polyomaviruses did not form monophyletic groups. Using our best hypothesis of polyomavirus evolutionary relationships and established host phylogenies, we performed a cophylogenetic reconciliation analysis of codivergence. Our analyses generated six optimal cophylogenetic scenarios of coevolution, including 12 codivergence events (P < 0.01), suggesting that Polyomaviridae coevolved with their avian and mammal hosts. As individual lineages, our analyses showed evidence of host switching in four terminal branches leading to MPtV, bovine polyomavirus, Sa12, and BK virus, suggesting a combination of vertical and horizontal transfer in the evolutionary history of the polyomaviruses.     

Comparing and predicting between several methods of measurement

In studies designed to compare different methods of measurement where more than two methods are compared or replicate measurements by each method are available, standard statistical approaches such as computation of limits of agreement are not directly applicable. A model is presented for comparing several methods of measurement in the situation where replicate measurements by each method are available. Measurements are viewed as classified by method, subject and replicate. Models assuming exchangeable as well as non-exchangeable replicates are considered. A fitting algorithm is presented that allows the estimation of linear relationships between methods as well as relevant variance components. The algorithm only uses methods already implemented in most statistical software.     

Efficiency of sunlight utilization: tubular versus flat photobioreactors

The light saturation effect imposes a serious limitation on the efficiency with which solar energy can be utilized in outdoor algal cultures. One solution proposed to reduce the intensity of incident solar radiation and overcome the light saturation effect is spatial dilution of light (i.e., distribution of the impinging photon flux on a greater photosynthetic surface area), but consistent experimental data supporting a significant positive influence of spatial light dilution on the productivity and the photosynthetic efficiency of outdoor algal cultures have never been reported. We used a coiled tubular reactor and compared a near-horizontal straight tubular reactor and a near-horizontal flat panel in outdoor cultivation of the cyanobacterium Arthrospira (Spirulina) platensis under defined operating conditions for optimum productivity. The photosynthetic efficiency achieved in the tubular systems was significantly higher because their curved surface diluted the impinging solar radiation and thus reduced the light saturation effect. This interpretation was supported by the results of experiments carried out in the laboratory under continuous artificial illumination using both a flat and a curved chamber reactor. The study also showed that, when the effect of light saturation is eliminated or reduced, productivity and solar irradiance are linearly correlated even at very high diurnal irradiance values, and supported findings that outdoor algal cultures are light-limited even during bright summer days. It was also observed that, besides improving the photosynthetic efficiency of the culture, spatial dilution of light also leads to higher growth rates and lowers the cellular content of accessory pigments; that is, it reduces mutual shading in the culture. The inadequacy of using volumetric productivity as the sole criterion for comparing reactors of different surface-to-volume ratio and of the areal productivity for evaluating the performance of elevated photobioreactors operated outdoors is stressed; it is furthermore suggested that the photosynthetic efficiency achieved by the culture also be calculated to provide a suitable parameter for comparison of different algal cultivation systems operated under similar climatic conditions. Copyright 1998 John Wiley & Sons, Inc.     

Comparing the MicroRNA Spectrum between Serum and Plasma

MicroRNAs (miRNAs) are small, non-coding RNAs that regulate various biological processes, primarily through interaction with messenger RNAs. The levels of specific, circulating miRNAs in blood have been shown to associate with various pathological conditions including cancers. These miRNAs have great potential as biomarkers for various pathophysiological conditions. In this study we focused on different sample types' effects on the spectrum of circulating miRNA in blood. Using serum and corresponding plasma samples from the same individuals, we observed higher miRNA concentrations in serum samples compared to the corresponding plasma samples. The difference between serum and plasma miRNA concentration showed some associations with miRNA from platelets, which may indicate that the coagulation process may affect the spectrum of extracellular miRNA in blood. Several miRNAs also showed platform dependent variations in measurements. Our results suggest that there are a number of factors that might affect the measurement of circulating miRNA concentration. Caution must be taken when comparing miRNA data generated from different sample types or measurement platforms.