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IYXX. Oxygen limitation of N, fixation in stem-girdled and nitrate-treated soybean. -Physiol. Plant. 73: 11S121.The effects of increasing rhizosphere PO, on nitrogenase activity and nodule resistance to O2 diffusion were investigated in soybean plants [Glycine max (L.) Merr. cv. Harosoy 631 in which nitrogenase (EC 1.7.99.2) activities were inhibited by (a) removal of the phloem tissue at the base of the stem (stem girdling), (b) exposure of roots to 10 mM NOj over 5 days (NOj-treated). or (c) partial inactivation of nitrogenase activity by an exposure of nodulated roots to 100 kPa O2 (O?-inhibited). In control plants and in plants which had been treated with 1 0 kPa 02, increasing rhizosphere 0, concentrations in 10 kPa increments from 20 to 70 kPa did not alter the steady-state nitrogenase activity. In contrast. in plants in which nitrogenase activities were depressed by stem girdling or by exposure to NOj. increasing rhizosphere p 0 2 resulted in a recovery of 57 or 67%. respectively. of the initial, depressed rates of nitrogenase activity. This suggests that the nitrogenase activity of stemgirdled and NO;-treated soybeans was O,-limited.O2 at 20 kPa and an apparent insensitivity of diffusion resistance to increases in external PO?.

The closed acetylene reduction assay has been used as a measure of nitrogenase activity and an indicator of N, fixation in Rhizobium/legume symbioses for 25 years. However, starting 10 years ago this assay has come under harsh criticism as being inaccurate. Currently, confusion exists regarding the conditions under which the acetylene reduction assay can be used accurately, or whether it can be used at all as a measure of nitrogenase activity. This article reviews the circumstance that has lead to this confusion. The author argues that under the proper assay conditions and with the appropriate checks, the closed acetylene reduction assay is still a valuable tool in assessing relative differences in nitrogenase activity in Rhizobium/legume symbioses.

In the mutualistic symbioses between legumes and rhizobia, actinorhizal plants and Frankia, Parasponia sp. and rhizobia, and cycads and cyanobacteria, the N 2 -fixing microsymbionts exist in specialized structures (nodules or cyanobacterial zones) within the roots of their host plants. Despite the phylogenetic diversity among both the hosts and the microsymbionts of these symbioses, certain developmental and physiological imperatives must be met for successful mutualisms. In this review, phylogenetic and ecological aspects of the four symbioses are first addressed, and then the symbioses are contrasted and compared in regard to infection and symbio-organ development, supply of carbon to the microsymbionts, regulation of O 2 flux to the microsymbionts, and transfer of fixed-N to the hosts. Although similarities exist in the genetics, development, and functioning of the symbioses, it is evident that there is great diversity in many aspects of these root-based N 2 -fixing symbioses. Each symbiosis can be admired for the elegant means by which the host plant and microsymbiont integrate to form the mutualistic relationships so important to the functioning of the biosphere.

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To study the effect of root-zone pH on characteristic responses of NH4+ -fed plants, soybeans (Glycine max¿L.¿ Merr. cv. Ransom) were grown in flowing solution culture for 21 d on four sources of N (1.0 mol m-3 NO3-, 0.67 mol m-3 NO3- plus 0.33 mol m-3 NH4+, 0.33 mol m-3 NO3- plus 0.67 mol m-3 NH4+, and 1.0 mol m-3 NH4+) with nutrient solutions maintained at pH 6.0, 5.5, 5.0, and 4.5. Amino acid concentration increased in plants grown with NH4+ as the sole source of N at all pH levels. Total amino acid concentration in the roots of NH4+ -fed plants was 8 to 10 times higher than in NO3(-)-fed plants, with asparagine accounting for more than 70% of the total in the roots of these plants. The concentration of soluble carbohydrates in the leaves of NH4+ -fed plants was greater than that of NO3(-)-fed plants, but was lower in roots of NH4+ -fed plants, regardless of pH. Starch concentration was only slightly affected by N source or root-zone pH. At all levels of pH tested, organic acid concentration in leaves was much lower when NH4+ was the sole N source than when all or part of the N was supplied as NO3-. Plants grown with mixed NO3- plus NH4+ N sources were generally intermediate between NO3(-)- and NH4+ -fed plants. Thus, changes in tissue composition characteristic of NH4+ nutrition when root-zone pH was maintained at 4.5 and growth was reduced, still occurred when pH was maintained at 5.0 or above, where growth was not affected. The changes were slightly greater at pH 4.5 than at higher pH levels.

Treating seed with Bacillus cereus strain UW85 stimulates nodulation in soybean, but the underlying mechanisms of this stimulation are poorly understood. In this study we assessed the effects of inoculation on nodulation, nitrogenase activity and dry matter partitioning in soybean (cv. Maple Ridge), common bean (cv. OAC Rico) and pea (cv. Express) under controlled, gnotobiotic conditions. Plants were grown for 34 d under controlled-environment conditions without a mineral N source, at low two levels of (brady)rhizobia inoculation. Soybean and common bean were grown at a single temperature regime and pea was grown at two temperature regimes. Each trial consisted of five treatments (noninoculated control, low (brady)rhizobia inoculation, low (brady)rhizobia plus UW85 inoculation, high (brady)rhizobia inoculation, and high (brady)rhizobia plus UW85 inoculation) with six plants as replicates per treatment in a completely randomized design. Inoculation of soybean with UW85 increased growth of roots, shoots, and nodules across both levels of bradyrhizobia inoculation and increased plant N accumulation by 12%. In oculation with UW85 also increased whole-plant nodulation (nodules plant-1) by 16%, but there were no effects on specific nodulation [nodules g-1 root dry weight (DW)], individual nodule DW, nitrogenase activity (μmol H2 g-1 nodule DW h-1) or N2 fixation efficiency (g plant-N g-1 nodule DW). Inoculation of pea with UW85 at the low temperature regime increased whole-plant nodulation by 19%, but had no positive effects on any other growth parameters. Inoculation of common bean with UW85 had no positive effects on any aspects of growth, nodulation or nitrogenase activity. Our data indicate that under gnotobiotic conditions, UW85 inoculation increases nodulation in soybean indirectly by increasing root growth and not stimulating the nodulation process per se. The bacterium had little to no positive effects on pea and common bean symbioses. Key words: Bacillus cereus UW85, Glycine max, growth-promotion, nodulation, Phaseolus vulgaris, Pisum sativum

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