Roots and Their Symbiotic Microbes: Strategies to Obtain Nitrogen and Phosphorus in a Nutrient-Limiting Environment

The association between Rhizobium and legumes and that between arbuscular mycorrhizal (AM) fungi and most land plants display a remarkable degree of similarity. Both events involve the recognition of, entrance into, and coexistence within the plant root, with the development of a specialized interface that always separates the two partners and at which nutrient exchange occurs. Molecules produced by rhizobia during the early stages of the symbiosis are related to fungal chitin, and the plant responds to both microbes with an increase in the production of flavonoids, which may assist in recognition and development of the symbioses. Many of the same plant genes are up-regulated in the two symbiotic pathways, and notably plants that are Nod^– are often defective in the AM association as well. However, there are a number of differences between the associations, and these are important for understanding the relationship between the two symbioses. The Rhizobium and AM symbioses will be compared and the question of whether the nitrogen-fixing association evolved from the much more ancient AM symbiosis will be discussed.     

Soil Phosphorus Fractions and Symbiotic Nitrogen Fixation across a Substrate-Age Gradient in Hawaii

We evaluated soil phosphorus (P) fractions, other soil characteristics, and rates of symbiotic N₂ fixation across a substrate-age gradient in Hawaii that was dominated by the leguminous tree Acacia koa (koa). Patterns of soil P observed on this gradient were compared to those on a slightly wetter gradient dominated by the nonfixer Metrosideros polymorpha (ohia). Along both gradients, concentrations of primary-mineral P fell sharply between the young and intermediate-aged sites, while labile inorganic P declined most steeply between the intermediate-aged and old sites. The most marked difference between the two gradients was that total soil carbon (C), nitrogen (N), and P, as well as nonoccluded organic P, were more variable across the ohia gradient, increasing to the intermediate-aged sites, then declining sharply at the old site. On the koa gradient, specific nitrogenase activity, measured by the acetylene-reduction (AR) assay, decreased three- to eightfold between the young site and the intermediate-aged and old sites, respectively. Nodule biomass showed no clear pattern. N₂ fixation rates, estimated by combining AR activity and nodule biomass measurements, were up to 8 kg N · ha⁻¹ · y⁻¹ at the young site and no more than 2 kg N · ha⁻¹ · y⁻¹ at the older sites, suggesting that koa may be a modest source of N in these Hawaiian forests. Received 26 September 2000; accepted 15 February 2002     

Tissue Nitrogen and Phosphorus in Seaweeds in a Tropical Eutrophic Environment: What a Long-Term Study Tells Us

Percentages of nitrogen and phosphorus in 10 species of seaweeds (6 green and 4 red algae) were monitored from 1997 to 2004 by seasonal sampling in Guanabara Bay, South-eastern Brazil. The species did not show consistent variations in tissue N, P and N:P that related to annual cycles. Throughout this study, higher percentages of tissue N and P were found in Bostrychia radicans and Grateloupia doryphora (red algae) and lower in Cladophora rupestris and Codium decorticatum (green algae). In November 1999, the Icaraí Submarine Sewage Outfall became operational, resulting in a reduction of visual pollution in the area and an improvement in the local quality of seawater for recreational use. Measurements of dissolved nutrients at the sampling site did not indicate significant changes in concentrations after the commissioning of the submarine sewage outfall; however, tissue P and N:P ratio of most of species were significantly lower than in the first two years of this survey. Variations in tissue nitrogen throughout this study were not significant, except for G. doryphora in some comparisons. Results show that seaweeds function very well as monitors of environmental changes in Guanabara Bay. Experimental data are needed to identify possible environmental processes which are promoting changes in chemical composition of the local seaweed populations.     

氮磷添加对麦冬根部养分浓度及其化学计量比的影响

以城市地被植物麦冬(Ophiopogon japonicus(Thunb.)Ker-Gawl.)为研究对象,研究了土壤中添加氮(N)磷(P)后对植物根部N、P养分及其化学计量比的影响。结果表明,实验监测期间(10-12月),麦冬根部N浓度平均值表现为N(5gm-2) P(1gm-2)处理>N(5gm-2)处理=P(1gm-2)处理>对照,根部P浓度和N:P比差异不显著(P>0.1)。而监测期间的N、P月份动态结果表明,同对照相比,N处理、P处理和N P处理的麦冬根部N、P浓度和N:P比的差异性均表现为监测前期(10月)较大,中后期(11-12月)较小的变化趋势。这说明麦冬能保持其根部N、P水平的稳定性,具有较强的应对N沉降的能力,且补充P肥可增强这种能力。因此,麦冬可在大气沉降严重的地区应用和推广。     

蕨类植物碳氮磷化学计量特征及其与土壤养分的关系

为探讨蕨类植物碳氮磷化学计量特征与土壤养分的关系,对福建省亚热带森林林下芒萁和乌毛蕨地上部分和地下部分的碳、氮、磷(C、N、P)含量和0~10 cm和10~20 cm两个土层的养分含量进行了测定。结果表明,无论是芒萁还是乌毛蕨,地上部分的N、P含量均高于地下部分,而C含量则无显著差异,导致地上部分的C∶N和C∶P均低于地下部分。与乌毛蕨相比,芒萁地上部分的N、P含量更低,地上和地下部分的C含量、C∶N和C∶P以及N、P含量的变异系数和表型可塑性指数则更高,表明芒萁采取了较高的养分利用效率和"表现最大化"的策略,而乌毛蕨则选择了较低的养分利用效率和"表现维持"的方式。两种蕨类植物地上和地下部分的N含量与土壤N含量(0~20 cm)均无显著相关。芒萁两个部位的P含量则均与土壤P含量(0~10 cm和10~20 cm)呈显著正相关,乌毛蕨P含量总体上与土壤P含量的相关性不显著(除地下部分的P含量与10~20 cm土层的P含量呈弱的正相关外)。这表明芒萁具有作为亚热带森林土壤P库指示植物的潜力。     

好氧反硝化微生物多样性及其反硝化功能初步研究

为研究污水厂/养殖池中好氧反硝化微生物的多样性及菌株反硝化能力,本研究采集了位于福建省厦门市和漳州市的污水处理厂、排污口、污水池、对虾养殖池的污水和污泥样品进行好氧反硝化微生物的富集、分离、鉴定和功能筛选。分别以NaNO3、NaNO2作为唯一氮源共分离纯化获得128株单菌。其中以NaNO3为唯一氮源分离得到63株,以NaNO2为唯一氮源分离得到65株。16SrRNA基因序列分析表明,128株单菌分属于γ-变形菌纲(Gammaproteobacteria,58.6%)、芽胞杆菌纲(Bacilli,6.4%)、放线菌纲(Actinobacteria,11.7%)、α-变形菌纲(Alphaproteobacteria,8.6%)、纤维菌纲(Cytophagia,2.3%)、鞘脂杆菌纲(Sphingobacteria,0.8%)和黄杆菌纲(Flavobacteria,1.6%)7个纲中的38个属。其中盐单胞菌属(Halomonas,29.7%)和芽胞杆菌属(Bacillus,12.5%)为优势菌属,并且广泛存在于各个样品中。反硝化功能初筛结果表明,35株菌能在72h内将20mg·L-1 NO-3-N/NO-2-N完全去除;复筛结果表明,21株菌能在72h内将100 mg·L-1 NO-3-N/NO-2-N完全去除,并且盐单胞菌属、卓贝尔氏菌属(Zobellella)、斯塔普氏菌属(Stappia)及节杆菌属(Arthrobactor)反硝化效果较好,其中斯塔普氏属是首次报道具有好氧反硝化功能。本研究结果表明,污水场/养殖池等环境中可培养反硝化细菌多样性丰富,同时高效反硝化菌的获得也为含氮废水的生物处理提供了良好的菌种资源。     

桉树根系共生真菌的分离与鉴定

南方土壤缺磷现象较为严重,菌根真菌等共生真菌对植物吸收磷等养分具有重要的促进作用。该研究采集尾叶桉(Eucalyptus urophylla)、窿缘桉(Eucalyptus exserta)和尾巨桉(E.urophylla×E.grandis)3种华南地区主要造林树种根系,采用组织分离法进行真菌分离,通过形态特征和核糖体18SrDNA基因ITS序列分析进行鉴定,经柯赫氏法则(Kochs Rule)确定桉树根系共生真菌,为桉树共生真菌理论研究和资源利用提供依据。结果表明:(1)3种桉树根系中,窿缘桉具有外生菌根(ectomycorrhizas,ECM)和丛枝菌根(arbuscular mycorrhizas,AM)结构,尾叶桉和尾巨桉同时具有AM结构、ECM结构和深色有隔内生真菌(dark septate endophytes fungi,DSE)结构。(2)3种桉树根系中分离鉴定出6种真菌:三色小皮伞菌(Marasmius tricolor)、黑柄裸脚伞(Gymnopus melanopus)、茎点霉属(Phomasp.)、镰刀霉属(Fusariumsp.)、二型伞霉(Umbelopsis dimorpha)和芒弗里亚拟盘多毛孢(Pestalotiopsis mangifolia)。(3)6种真菌回接巨桉(Eucalyptus grandis)组培苗,三色小皮伞菌和黑柄裸脚伞形成ECM结构,为ECM真菌;茎点霉属、镰刀霉属、二型伞霉和芒弗里亚拟盘多毛孢形成DSE典型的深色有隔菌丝和微菌核结构,推测为DSE;其中2种ECM真菌为桉树中首次报道。     

Focus Issue on Roots: The Optimal Lateral Root Branching Density for Maize Depends on Nitrogen and Phosphorus Availability

Observed phenotypic variation in the lateral root branching density (LRBD) in maize (Zea mays) is large (1–41 cm⁻¹ major axis [i.e. brace, crown, seminal, and primary roots]), suggesting that LRBD has varying utility and tradeoffs in specific environments. Using the functional-structural plant model SimRoot, we simulated the three-dimensional development of maize root architectures with varying LRBD and quantified nitrate and phosphorus uptake, root competition, and whole-plant carbon balances in soils varying in the availability of these nutrients. Sparsely spaced (less than 7 branches cm⁻¹), long laterals were optimal for nitrate acquisition, while densely spaced (more than 9 branches cm⁻¹), short laterals were optimal for phosphorus acquisition. The nitrate results are mostly explained by the strong competition between lateral roots for nitrate, which causes increasing LRBD to decrease the uptake per unit root length, while the carbon budgets of the plant do not permit greater total root length (i.e. individual roots in the high-LRBD plants stay shorter). Competition and carbon limitations for growth play less of a role for phosphorus uptake, and consequently increasing LRBD results in greater root length and uptake. We conclude that the optimal LRBD depends on the relative availability of nitrate (a mobile soil resource) and phosphorus (an immobile soil resource) and is greater in environments with greater carbon fixation. The median LRBD reported in several field screens was 6 branches cm⁻¹, suggesting that most genotypes have an LRBD that balances the acquisition of both nutrients. LRBD merits additional investigation as a potential breeding target for greater nutrient acquisition.At least four major classes of plant roots can be distinguished based on the organ from which they originate: namely the seed, the shoot, the hypocotyl/mesocotyl, and other roots (Zobel and Waisel, 2010). The last class is lateral roots, which form in most plants the majority of the root length, but not necessarily of the root weight, as lateral roots have smaller diameter. Lateral roots start with the formation of lateral root primordia, closely behind the root tip of the parent root. These primordia undergo nine distinguishable steps, of which the last step is the emergence from the cortex of the parent root just behind the zone of elongation, usually only a few days after the first cell divisions that lead to their formation (Malamy and Benfey, 1997). However, not all primordia develop into lateral roots; some stay dormant (Dubrovsky et al., 2006), although dormancy of primordia may not occur in maize (Zea mays; Jordan et al., 1993; Ploshchinskaia et al., 2002). The final number of lateral roots is thereby dependent on the rate of primordia formation as well as the percentage of primordia that develop into lateral roots. This process of primordia formation and lateral root emergence is being studied intensively, including the genes that are activated during the different steps and the hormones regulating the process (López-Bucio et al., 2003; Dubrovsky et al., 2006; Osmont et al., 2007; Péret et al., 2009; Lavenus et al., 2013). Significant genotypic variation in the density of lateral roots has been observed, ranging from no lateral roots to 41 roots cm⁻¹ in maize (Trachsel et al., 2010; Lynch, 2013). This suggests that clear tradeoffs exist for the development of lateral roots and that these genotypes have preprogrammed growth patterns that are adaptive to specific environments. While some of the variation for lateral root branching density (LRBD) that has been observed across environments, for example by Trachsel et al. (2010), is constitutive, many genotypes have strong plasticity responses of LRBD to variations in soil fertility (Zhu et al., 2005a; Osmont et al., 2007). Both the nutrient and carbon status of the plant and the local nutrient environment of the (parent) root tip influence LRBD. Many studies have documented these plasticity responses, and others have tried to unravel parts of the sensing and signaling pathways that regulate LRBD. The utility of root proliferation into a nutrient patch has been studied and debated (Robinson et al., 1999; Hodge, 2004), but much less so the utility of having fewer or more branches across the whole root system. Our understanding of the adaptive significance of variation in LRBD among genotypes is thereby limited, with many studies not accounting for relevant tradeoffs. In this study, we integrate several functional aspects of LRBD with respect to nutrient acquisition, root competition, and internal resource costs and quantify these functional aspects using the functional structural plant model SimRoot. SimRoot simulates plant growth with explicit representation of root architecture in three dimensions (Fig. 1; Supplemental Movie S1). The model focuses on the resource acquisition by the root system and carbon fixation by the shoot while estimating the resource utilization and requirements by all the different organs.

Table I.

Minimum, maximum, and median LRBD in different populations phenotyped by various researchers at several locations in the worldLocations are as follows: D, Juelich, Germany; PA, State College, PA; and SA, Alma, South Africa. Some of the experiments included nutrient treatments: LN, low nitrogen availability; and LP, low phosphorus availability. Data were collected by counting the number of roots along a nodal root segment. Data were supplied by the person indicated under source: H.S., H. Schneider; L.Y., L. York; A.Z., A. Zhan; and J.P., J.A. Postma. WiDiv, Wisconsin Diversity panel; IBM, intermated B73 × Mo17; NAM, nested association mapping.

How Microbes Can Achieve Balanced Growth in a Fluctuating Environment

A microbial colony needs several essential nutrients in order to grow. Moreover, the colony requires these nutrients in fixed combinations, which are dictated by the chemical composition of its biomass. Unfortunately, ambient availabilities of the various nutrients vary all the time. This poses the question of how microbes can achieve balanced growth.The present solution to this problem is novel in that the allocation of molecular building blocks among assimilatory machineries within the cell is regarded as dynamic. This paper shows that allocation can be adapted so as to achieve balanced growth, nearly regardless of environmental conditions. Moreover, it is shown that a feedback mechanism, which monitors internal stores, is able to achieve this allocation.     

一种高效研究大豆根瘤共生固氮的营养液栽培体系

为建立一种既可高效结瘤固氮, 又具有一定产量的大豆(Glycine max)营养液栽培系统, 设计并进行了2个试验。首先在不同供氮条件下, 研究了接种根瘤菌对大豆的结瘤状况、固氮能力、生物量及产量的影响。结果表明, 供氮过高或过低, 均影响大豆生长、产量形成及根瘤固氮; 并且植物生长所需的最适供氮水平远高于生物固氮所需的最适供氮水平。此外, 大豆生物固氮活性最高的时期在生殖期第一期(R1期)之前。由此推断, 大豆R1期前, 供应较低的氮, 有利于根瘤形成及固氮; 而从R1期起, 应提高供氮水平, 以促进植物生长及产量的形成。在此基础上开展第2个试验, 对供氮条件进行了优化处理(即R1期前低氮供应、R1期开始中氮供应)。结果表明, 与持续供应高氮相比, 优化供氮处理不仅可获得较多固氮酶活性较高的大根瘤, 还能保持较好的生长、获得更高的百粒重及维持80%左右的产量。研究结果不仅可为高效研究大豆根瘤共生固氮提供营养液配方, 还可为大豆高产高效栽培提供试验依据。     

黄芩干物质累积和氮磷钾吸收、分配规律研究

为了探讨黄芩干物质累积和氮、磷、钾吸收与分配的特点及两者间的相互关系,通过田间试验和采样分析,研究了黄芩不同生育期植株的干物质和氮、磷、钾累积量.结果表明,黄芩干物质的累积量随生育进程不断地增加,出苗后52～85 d干物质累积量占总累积量的61.62％.在整个生育期,黄芩对K2O的吸收累积量最大,N次之,P2O5最小,N、P2O5、K2O吸收比例约为2.8∶1.0∶2.9,并且黄芩地上部氮磷钾的累积量大于根部,不同生育期,根部N、P2O5、K2O的累积比例呈现增加—降低—增加的趋势.黄芩对氮磷钾的积累量与干物质积累量呈极显著正相关关系.在供试的土壤和施肥条件下,每生产100 kg的黄芩根需要从土壤和肥料中吸收6.34 kg的N,2.60 kg的P2O5,7.02 kg的K2O.     

Production of the Phytoalexin Glyceollin I by Soybean Roots in Response to Symbiotic and Pathogenic Infection

The amount of the phytoalexin glyceollin I in root exudate and root hairs of individual seedlings of Glycine max (L. Merr. cv. Preston) was analysed using a radioimmunoassay. Bradyrhizobium japonicum 110spc4, which is able to form nitrogen fixing nodules with this plant, caused an increase of up to 50-fold in glyceollin I levels in root exudate relative to uninfected control seedlings. Maximum glyceollin I levels were reached within 10 h of incubation. Elevated glyceollin I levels were also observed after incubation of soybean roots in sterile bacterial supernatant, a suspension of autoclaved bacteria or the supernatant from broken cells of Bradyrhizobium japonicum. Increased glyceollin I production is not due to the process of active root hair penetration by the microsymbiont since living bacterial cells are not necessary for the induction. The observed glyceollin I production in response to Bradyrhizobium japonicum is several times lower than that after pathogenic infection. Infection with zoospores of the phytopathogenic oomycete, Phytophthora megasperma f. sp. glycinea race 1, leads within 20 h to an accumulation of 7 nmol glyceollin I/seedling in the root exudate of the compatible cultivar Kenwood and 48 nmol glyceollin I/seedlings in that of the incompatible cultivar Maple Arrow. These results support the idea that phytoalexins are implicated in determination of compatibility in pathogenic interactions. Crude cell extracts of different symbiotic bacteria (Bradyrhizobium japonicum 110spc4, Rhizobium meliloti 2011, Rhizobium leguminosarum PRE 8, Sinorhizobium fredii HH 103) were found to induce different amounts of glyceollin I in the root exudate. The observed glyceollin I levels could not be correlated with the ability of these rhizobial strains to nodulate Glycine max. Inhibition of flavonoid and phytoalexin synthesis by (R)-(1-amino-2-phenylethyl)phosphonic acid (APEP), a specific inhibitor of the phenylalanine-ammonia-lyase (PAL), during the first 20 h of the symbiotic interaction dramatically decreased the number of nodules formed in root regions that had been in contact with the inhibitor. This effect was observed at concentrations that inhibited neither bacterial nor plant growth. The implications of these findings for the process of nodule initation are discussed.     

Halotropism Is a Response of Plant Roots to Avoid a Saline Environment

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Harmonizing Labeling and Analytical Strategies to Obtain Protein Turnover Rates in Intact Adult Animals

Changes in the abundance of individual proteins in the proteome can be elicited by modulation of protein synthesis (the rate of input of newly synthesized proteins into the protein pool) or degradation (the rate of removal of protein molecules from the pool). A full understanding of proteome changes therefore requires a definition of the roles of these two processes in proteostasis, collectively known as protein turnover. Because protein turnover occurs even in the absence of overt changes in pool abundance, turnover measurements necessitate monitoring the flux of stable isotope–labeled precursors through the protein pool such as labeled amino acids or metabolic precursors such as ammonium chloride or heavy water. In cells in culture, the ability to manipulate precursor pools by rapid medium changes is simple, but for more complex systems such as intact animals, the approach becomes more convoluted. Individual methods bring specific complications, and the suitability of different methods has not been comprehensively explored. In this study, we compare the turnover rates of proteins across four mouse tissues, obtained from the same inbred mouse strain maintained under identical husbandry conditions, measured using either [¹³C₆]lysine or [²H₂]O as the labeling precursor. We show that for long-lived proteins, the two approaches yield essentially identical measures of the first-order rate constant for degradation. For short-lived proteins, there is a need to compensate for the slower equilibration of lysine through the precursor pools. We evaluate different approaches to provide that compensation. We conclude that both labels are suitable, but careful determination of precursor enrichment kinetics in amino acid labeling is critical and has a considerable influence on the numerical values of the derived protein turnover rates.     

白云山土壤微生物的季节变化及其对环境污染的反应

初步调查了广州市白云山土壤中细菌、放线菌和真菌等微生物．在人为活动频繁的样地土壤微生物的多样性及其生物量均大于森林内的３个样地．土壤微生物的生物量高峰期出现在秋冬季节．用不同污染程度的水处理土壤１５ｄ后,以蒸馏水（ｐＨ７．０）处理为对照,土壤微生物种类多样性的顺序为：蒸馏水＞溪水（ｐＨ５．４）＞雨水（ｐＨ４．８）＞污水（ｐＨ４．２）;而土壤微生物的生物量则无显著变化     

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.     

Symbiotic Nitrogen Fixation and Ammonium Nitrogen Assimilation in rrrbrb-, rrRbRb-, and RRrbrb-Pea Mutants

Wrinkled-seeded pea mutants (Pisum sativum L., genotypes rrrbrb-, rrRbRb-, and RRrbrb-) have seeds with reduced, but different, starch content and modified starch properties. Analysis of these mutants revealed an enhanced capacity of root nodules for symbiotic nitrogen fixation and of host plant organs for assimilation of ammonium nitrogen. This observation was confirmed by morphological data on organization of symbiotic system, by elevated nitrogenase activity, high protein accumulation in plants due to nitrogen fixation, and by enhanced activity of glutamine synthase in leaves and glutamate dehydrogenase in roots of mutants, as compared with the organs of wild-type pea. It is supposed that the aforementioned advantages of mutants are related to accumulation in seeds of elevated protein reserves that satisfy their demand for nitrogen during formation of symbiotic systems.