Effects of calcium deficiency on symbiotic nitrogen fixation

The mechanism of the effect of mild calcium deficiency on nitrogen metabolism of the symbiotic plant was studied from the distribution of calcium and of nitrogen and carbohydrate fractions in plant organs.

Nitrogen concentrations of all plant organs decreased with calcium deficiency. Addition of either a nitrogen or a calcium salt increased nitrogen concentrations. For roots as well as whole plants the effects of one salt were largely replaced by the other. These effects establish that calcium deficiency decreased the supply of fixed nitrogen from nodules to other organs. As weight of nodules was independent of calcium it follows that nodular efficiency was impaired.

Since nitrogen concentrations of nodules decreased rather than increased with calcium deficiency it is suggested that calcium deficiency interferes with the rate of reduction of nitrogen in the nodule rather than with the export of reduced nitrogen. Distribution of α-amino, amide, and ammonium nitrogen are consistent with this suggestion.

Nitrogen fixation was not limited by translocation of carbohydrates to nodules as calcium deficiency had little effect on the concentration of soluble carbohydrates and actually increased the concentration of starch in nodules.

Calcium deficiency depressed the calcium content of nodules so that nitrogen fixation may have been impaired by inadequate calcium for nodular structure or metabolism.

     

Effects of drought on nitrogen fixation in soybean root nodules

Soybean plants [Glycine max (L.) Merr.] were grown in silica sand and were drought stressed for a 4 week period during reproductive development and without any mineral N supply in order to maximize demand for fixed nitrogen. A strain of Bradyrhizobium japonicum that forms large quantities of polysaccharide in nodules was used to determine whether or not the supply of reduced carbon to bacteroids limits nitrogenase activity. A depression of 30–40% in nitrogen content in leaves and pods of stressed plants indicated a marked decline in nitrogen fixation activity during the drought period. A 50% increase in the accumulation of bacterial polysaccharide in nodules accompanied this major decrease in nitrogen fixation activity and this result indicates that the negative impact of drought on nodules was not due to a depression of carbon supply to bacteroids. The drought treatment resulted in a statistically significant increase in N concentration in leaves and pods. Because N concentration and chlorophyll concentration in leaves were not depressed, there was no evidence of nitrogen deficiency in drought‐stressed plants, and this result indicates that the negative impact of drought on nodule function was not the cause of the depression of shoot growth. At the end of the drought period, the concentration of carbohydrates, amino nitrogen, and ureides was significantly increased in nodules on drought‐stressed plants. The overall results support the view that, under drought conditions, nitrogen fixation activity in nodules was depressed because demand for fixed N to support growth was lower.     

Bacteria-plant interactions in symbiotic nitrogen fixation

In the inter- and intracellular N₂-fixing symbioses between plants and micro-symbionts, the development of an endophytic form of the micro-symbiont is essential. This development includes a series of steps consisting of plant-bacteria interactions. Considerable progress in the elucidation of these steps has been made by applications of the methods of molecular genetics. Several genes with a role during infection and nodulation have been indicated in Rhizobium and Bradyrhizobium like the common nod genes A, B, C, I and J, and the host-specific genes nod E, F and H. The nod D gene is the only constitutive gene, and its product is essential for activity of all other nod genes, provided some flavonoids from the root exudate are present as well. Mutants in these genes show phenotypic effects, in which the products of the genes must be involved. Far more difficult is the biochemical and physiological study of these products and their direct effects. The difficulties involved in such biochemical-physiological studies is illustrated by a short discussion of the controversies around the possible role of plant lectins. While in Rhizobium the nod genes are present on a large sym-plasmid, other essential genes must be present on the bacterial chromosome and on other plasmids. Induction of plant genes is evident from the formation of nodule-specific proteins, the nodulins. Though many different plant and bacterial genes are involved in the series of steps in the development of an effective root nodule, there are indications that regulation is affected by a smaller number of essential regulatory genes. This is illustrated by the effect of the regulatory nod D gene during infection and nodulation, and of ntrA and nifA genes for the formation and activation of the nitrogen-fixing systems. Moreover, every step, once initiated, may lead to cascade effects on subsequent reactions. Finally, some further consequences of the endophytic way of life are discussed, which affect either the metabolic and transport activities of the endophytes or their viability. This is illustrated by the possible role of membrane integrity as evident during the isolation of Frankia from its endophytic form.     

Genomics insights into symbiotic nitrogen fixation

Following an interaction with rhizobial soil bacteria, legume plants are able to form a novel organ, termed the root nodule. This organ houses the rhizobial microsymbionts, which perform the biological nitrogen fixation process resulting in the incorporation of ammonia into plant organic molecules. Recent advances in genomics have opened exciting new perspectives in this field by providing the complete gene inventory of two rhizobial microsymbionts. The complete genome sequences of Mesorhizobium loti, the symbiont of several Lotus species, and Sinorhizobium meliloti, the symbiont of alfalfa, were determined and annotated in detail. For legume macrosymbionts, expressed sequence tag projects and expression analyses using DNA arrays in conjunction with proteomics approaches have identified numerous genes involved in root nodule formation and nitrogen fixation. The isolation of legume genes by tagging or positional cloning recently allowed the identification of genes that control the very early steps of root nodule organogenesis.     

Methods for enhancing symbiotic nitrogen fixation

Biological nitrogen fixation of leguminous crops is becoming increasingly important in attempts to develop sustainable agricultural production. However, these crops are quite variable in their effectiveness in fixing nitrogen. By the use of the ¹⁵N isotope dilution method some species have been found to fix large proportions of their nitrogen, while others like common bean have been considered rather inefficient. Methods for increasing N₂ fixation are therefore of great importance in any legume work. Attempts to enhance nitrogen fixation of grain legumes has been mainly the domain of microbiologists who have selected rhizobial strains with superior effectiveness or competitive ability. Few projects have focused on the plant symbiont with the objective of improving N₂ fixation as done in the FAO/IAEA Co-ordinated Research Programme which is being reported in this volume. The objective of the present paper is to discuss some possibilities available for scientists interested in enhancing symbiotic nitrogen fixation in grain legumes. Examples will be presented on work performed using agronomic methods, as well as work on the plant and microbial symbionts. There are several methods available to scientists working on enhancement of N₂ fixation. No one approach is better than the others; rather work on the legume/Rhizobium symbiosis combining experience from various disciplines in inter-disciplinary research programmes should be pursued.     

Photosynthesis and nitrogen fixation in legume plants

Plants pass through a succession of growth phases at a rate largely controlled by environmental factors. The spatial arrangement and efficiency of plant organs are influenced by the fluxes of energy and matter in their environments. Thus, the successful integration of processes, such as photosynthesis and nitrogen fixation, occurring in the very different environments of the soil and the air requires a complex functional balance. Such a balance is particularly complex for legumes in which the genetic expressions of the host plant and Rhizobium influence the nitrogen economy. Progress towards improvements in symbiotic nitrogen fixation has been severely limited by the difficulty of distinguishing between the metabolic activities of the roots and nodules in whole plant studies. Recent improvements in experimental precision have revealed processes which govern gaseous diffusion in nodules and control their carbohydrate use. Furthermore, the application of quantitative models to problems of carbon and nitrogen nutrition is improving the understanding of plant growth.     

干旱胁迫对毛竹幼苗生理特性的影响

以1年生毛竹为试验材料,采用盆栽方式,设置对照(CK)、轻度(LS)、中度(MS)和重度干旱胁迫(HS)4个不同处理,研究不同水分条件下毛竹幼苗生理响应。结果表明:随着水分胁迫程度的加强,毛竹幼苗净光合速率(Pn)、蒸腾速率(Tr)显著下降(P<0.01),降幅分别达54.7%和49.7%,水分利用效率(WUE)在轻度胁迫条件下明显提高,但在中度和重度胁迫下又有一定程度下降;叶绿素a和叶绿素b含量显著下降(P<0.05和P<0.01),降幅分别为42.1%和28.2%,水分胁迫对叶绿素a/b(Chl-a/Chl-b)影响不显著;SOD活性、CAT活性和MDA含量显著上升(P<0.01、P<0.05和P<0.01);各生理指标间存在着相关性,经主成分分析,可将9个单项耐旱生理指标综合成为2个相互独立的综合指标,为评价毛竹苗期抗旱性提供参考。     

Legume haemoglobins: symbiotic nitrogen fixation needs bloody nodules

How do plants create an environment in which symbiotic bacteria can reduce enough N2 to provide the plant with sufficient ammonium for growth? Gene silencing has now been used to show that legume haemoglobins are crucial.     

Enhanced expression of Rhizobium etli cbb 3 oxidase improves drought tolerance of common bean symbiotic nitrogen fixation

To investigate the involvement of Rhizobium etli cbb (3) oxidase in the response of Phaseolus vulgaris to drought, common bean plants were inoculated with the R. etli strain, CFNX713, overexpressing this oxidase in bacteroids (cbb (3) (+)) and subjected to drought conditions. The negative effect of drought on plant and nodule dryweight, nitrogen content, and nodule functionality was more pronounced in plants inoculated with the wild-type (WT) strain than in those inoculated with the cbb (3) (+) strain. Regardless of the plant treatment, bacteroids produced by the cbb (3) (+) strain showed higher respiratory capacity than those produced by the WT strain. Inoculation of plants with the cbb (3) (+) strain alleviated the negative effect of a moderate drought on the respiratory capacity of bacteroids and the energy charge of the nodules. Expression of the FixP and FixO components of the cbb (3) oxidase was higher in bacteroids of the cbb (3) (+) strain than in those of the WT strain under all experimental conditions. The decline in sucrose synthase activity and the decrease in dicarboxylic acids provoked by moderate drought stress were more pronounced in nodules from plants inoculated with the WT strain than in those inoculated with the cbb (3) (+) strain. Taken together, these results suggest that inoculation of plants with a R. etli strain having enhanced expression of cbb (3) oxidase in bacteroids reduces the sensitivity of P. vulgaris-R. etli symbiosis to drought and can modulate carbon metabolism in nodules.     

Molecular genetics of Rhizobium Meliloti symbiotic nitrogen fixation

The application of recombinant DNA techniques to the study of symbiotic nitrogen fixation has yielded a growing list of Rhizobium meliloti genes involved in the processes of nodulation, infection thread formation and nitrogenase activity in nodules on the roots of the host plant, Medicago sativa (alfalfa). Interaction with the plant is initiated by genes encoding sensing and motility systems by which the bacteria recognizes and approaches the root. Signal molecules, such as flavonoids, mediate a complex interplay of bacterial and plant nodulation genes leading to entry of the bacteria through a root hair. As the nodule develops, the bacteria proceed inward towards the cortex within infection threads, the formation of which depends on bacterial genes involved in polysaccharide synthesis. Within the cortex, the bacteria enter host cells and differentiate into forms known as bacteroids. Genes which encode and regulate nitrogenase enzyme are expressed in the mature nodule, together with other genes required for import and metabolism of carbon and energy sources offered by the plant.     

干旱胁迫对大丽花生理生化指标的影响

以大丽花品种'粉西施'扦插苗为试材,研究不同程度干旱胁迫及复水处理对大丽花品种'粉西施'叶片生理生化指标的影响.结果表明:随着干旱胁迫程度的增加和时间的延长,大丽花叶片的相对含水量、水势和叶绿素含量显著下降;叶片相对电导率、丙二醛含量显著增加,质膜遭到损害,大量离子外渗,严重胁迫下质膜的损害最严重,复水后无法恢复到对照水平;可溶性糖、脯氨酸含量显著增加,脯氨酸含量在中度和重度胁迫后期增加显著,说明其对干旱亏缺的敏感性较低;可溶性蛋白呈降-升-降的变化趋势;抗氧化酶SOD、POD和CAT活性先上升后下降,3种酶对干旱胁迫和活性氧的响应存在一定差异,表现为相互协调的作用.