Effect of micronutrient application on the growth and occurrence of sterility in barley and rice in a Malaysian deep peat soil

Abstract

Nodule formation in legume crops is a multistep process which involves the interactive gene expression of bacteria and host plants. Attempts to widen the genetic variation of symbiotic bacteria and host plants have led to the isolation of several mutants with a diverse capability of nodulation. The most striking variants in hosts are the supernodulating mutants isolated in pea (Pisum sativum L.) (Jacobsen and Feenstra 1984; Duc and Messager 1989), common bean (Phaseolus vulgaris L.) (Park and Buttery 1988), and soybean (Glycine max (L.) Men.) (Carroll et al. 1985a,b; Gremaud and Harper 1989; Akao and Kouchi 1992), all of which are capable of producing several-fold more nodules than their parental lines in the presence of nitrate. These mutants may be useful materials for analyzing the mechanism controlling nodule formation, and are considered to have a high agronomic potential under certain growing conditions.     

Shoot control of nodulation is modified by the root in the supernodulating soybean mutant En6500 and its wild-type parent cultivar Enrei

Abstract

Based on grafting studies, both supernodulating (Carroll et al. 1985a, b) and hypernodulating (Gremaud and Harper 1989) soybean ([itGlycine max} L. Merr.) phenotypes were reported to be under the control of shoot factors (Delves et al. 1986, 1987; Cho and Harper 1991). Recently Akao and Kouchi (1992) have isolated a new supernodulating mutant (En6500) from ethyl methane sulfonate (EMS)-treated Enrei, a cultivar which is widely grown in the central districts of Japan. This mutant has been shown to produce several fold as many nodules as its wild-type parent cultivar when grown at a low concentration (0.5 mol m^-3) of nitrate. Moreover, it exhibited a continuous increase in the nodule number with the increasing nitrate concentration, even at 15 mol m^-3 (Francisco et al. 1992), in contrast to the similar mutant nts382 in which the nodulation decreased even at the relatively low nitrate level of 5.5 mol m^-3 (Carroll et al. 1985a). In this study we conducted grafting experiments to determine which plant part controls the supernodulation of En6500.     

硼对大豆结瘤和固氮作用的影响——Ⅰ.对生长、营养元素吸收与分布和固氮量影响

在短期无氮营养液栽培(28天)条件下,研究缺硼和正常供硼处理对Bragg大豆品种及不结瘤突变体nod49和超结瘤突变体nts382生长、矿质元素吸收与分布及固氮量的影响。试验结果表明:1.在正常供硼的条件下,不结瘤突变体nod的植株生长量,株高、主根生长均高于超结瘤突变体及其亲本。缺硼处理时三种基因型大豆的生长均受到明显的抑制作用。无论是正常供硼还是缺硼处理,超结瘤突变体nts382根系的生长量均明显小于其它基因型。2.正常供硼处理时三种基因型大豆地上部、根及nts382的根瘤中硼的浓度和积累量均高于缺硼处理。缺硼处理,超结瘤突变体及其亲本地上部和根中大部分矿质元素浓度高于正常供硼处理,但积累总量低于正常供硼处理或变化不大。施硼处理的nts382和Bragg,根和根瘤中钙的浓度和积累量明显降低,而地上部变化不大,同时在nts382根瘤中,锌、锰、铁、铜浓度和含量也下降,而钼含量增加。3.施硼处理和Bragg和nts382地上部、根及根瘤中氮的浓度和含量均显著高于缺硼处理,其中超结瘤突变体中氮浓度和总量最高。在缺硼条件下,Bragg和超结瘤突变体能够结瘤,但是Bragg大豆根瘤的固氮作用完全受到抑制,而nts382根瘤还能保持较强的固氮能力。     

Effect of Concentration of Phosphate on Spore Germination and Hyphal Growth of Arbuscular Mycorrhizal Fungus,Gigaspora margarita Becker & Hall

Extract

Arbuscular mycorrhizae are symbiotic associations between plant roots and soil fungi that belong to the order Glomales. They enhance the uptake of phosphate and improve plant growth (Mos.se 1973). Those functions have not fully been utilized in Japan, where a large amount of phosphate fertilizer is usually applied, because arbuscular mycorrhizal colonisation in plant is inhibited by a large application of phosphate (Mosse 1973; Abbott et al. 1984; Tawaraya et al. 1985; Schubert and Hayman 1986; Thomson et al. 1986; Amijee et al. 1989). It is important to analyse the conditions for symbiotic association between fungi and host plant to use arbuscular mycorrhiza.     

硼对大豆结瘤和固氮作用的影响——Ⅰ.对生长、营养元素吸收与分布和固氮量影响

在短期无氮营养液栽培(28天)条件下,研究缺硼和正常供硼处理对Bragg大豆品种及不结瘤突变体nod49和超结瘤突变体nts382生长、矿质元素吸收与分布及固氮量的影响。试验结果表明:1.在正常供硼的条件下,不结瘤突变体nod的植株生长量,株高、主根生长均高于超结瘤突变体及其亲本。缺硼处理时三种基因型大豆的生长均受到明显的抑制作用。无论是正常供硼还是缺硼处理,超结瘤突变体nts382根系的生长量均明显小于其它基因型。2.正常供硼处理时三种基因型大豆地上部、根及nts382的根瘤中硼的浓度和积累量均高于缺硼处理。缺硼处理,超结瘤突变体及其亲本地上部和根中大部分矿质元素浓度高于正常供硼处理,但积累总量低于正常供硼处理或变化不大。施硼处理的nts382和Bragg,根和根瘤中钙的浓度和积累量明显降低,而地上部变化不大,同时在nts382根瘤中,锌、锰、铁、铜浓度和含量也下降,而钼含量增加。3.施硼处理和Bragg和nts382地上部、根及根瘤中氮的浓度和含量均显著高于缺硼处理,其中超结瘤突变体中氮浓度和总量最高。在缺硼条件下,Bragg和超结瘤突变体能够结瘤,但是Bragg大豆根瘤的固氮作用完全受到抑制,而nts382根瘤还能保持较强的固氮能力。     

Plant growth,photosynthetic pigments,and nitrate assimilation in symbiotic mutants of garden pea

A set of 21 pea (Pisum sativum L.) non‐nodulating mutants and a supernodulating mutant were compared to the parental cv. ‘Finale’ by leaf nitrate reductase activity, nitrogen (N) content in the shoot, content of photosynthetic pigments, and growth. The plants were cultivated asymbiotically at the growth‐saturating nitrate (NO3) level (10 mM) to detect the nitrate assimilation faults. Both positive and negative deviations were revealed in all traits, indicating pleiotropy of symbiotic mutations and/or the deleterious effects of multiple mutations. Only four mutants were indistinguishable from the control plants by all traits. The supernodulating line RisfixC demonstrated lower shoot growth and increased N content in the shoot, in spite of the asymbiotic cultivation.     

The nature of non-protein nitrogen accumulation in Trifolium subterraneum root nodules

Non-protein nitrogen accumulated in nodules formed on Trifolium subterraneum cv. Tallarook by Rhizobium trifolii strain NA30, but not in nodules formed by strain TA1. Studies with six R. trifolii strains and four T. subterraneum cultivars indicated that the accumulation of non-protein nitrogen was a characteristic of certain strains and that it was accompanied by a greater development of nodule tissue. With normal symbiotic associations, approximately 6 per cent of the total plant nitrogen was located in the nodule system whereas nodules accumulating non-protein nitrogen contained, on average, 12 per cent of the total nitrogen in the plant.The principal component of the accumulating non-protein nitrogen was identified as “bound” γ-aminobutyric acid. Moderate to high concentrations of γ-aminobutyric acid (0.3–1.7 mmoles/g nodule dry weight) were found in nodules formed by 10 strains (of 36 examined) on Tallarook. With two “accumulating” strains, higher concentrations of γ-aminobutyric acid were found in nodules formed on the cultivar Clare (2.0 mmoles/g nodule dry weight) than in nodules formed on Tallarook or Yarloop (1 1.4 mmoles). No γ-aminobutyric acid was found in cultured cells of either an accumulating strain (NA30) or a nonaccumulating strain (TA1) of R. trifolii.The accumulation of non-protein nitrogen as γ-aminobutyric acid, and the accompanying increase in nodule tissue, each resulting in the export of a lower proportion of the nitrogen fixed to the host, is considered to be a factor causing a lower degree of symbiotic effectiveness.     

硼对大豆结瘤和固氮作用的影响——Ⅱ.对根瘤结构和固氮酶活性的影响

在短期无氮营养液栽培条件下研究了缺硼和正常供硼处理对大豆超结瘤突变体(nts382)及其亲本Bragg根瘤结构和根瘤固氮酶活性的影响。结果表明:1.在缺硼条件下,无论是超结瘤突变体nts382,还是亲本Bragg,其根瘤固氮酶活性都显著下降;2.无论是缺硼还是正常供硼处理,超结瘤突变体nts382单位鲜重根瘤活性都明显低于亲本,但由于nts382每株根瘤数和鲜重显著高于其亲本,使每株固氮酶活性明显高于亲本,植株中氮素积累总量也相应的高于亲本;3.缺硼条件下,根瘤细胞结构发生明显改变,细胞壁凸凹不平,细胞间隙加大,含菌细胞内类菌体含量减少。这可能是硼影响根榴细胞正常固氮功能的主要原因。     

Genistein addition to the rooting medium of soybean at the onset of nitrogen fixation increases nodulation

In the legume‐(Brady)Rhizobium symbiosis, signal exchange between the host‐plant and the symbiotic bacterium is an essential step in nodule formation. Genistein is the most effective plant‐to‐bacterium signal in the soybean [Glycine max (L.) Merr.] N₂‐fixing symbiosis. Its concentration in soybean root system increases with seedling development, and decreases immediately after the onset of N₂ fixation. This study was conducted to determine whether addition of genistein to the rooting medium at the onset of N₂ fixation would increase nodulation thereafter. The results indicated that watering soybean plants with a solution containing genistein beginning at the onset of N₂‐fixation increased nodule size, nodule number and nodule weight per plant. Shoot nitrogen (N) concentration was also increased. Soybean cultivar AC Bravor was more sensitive to genistein addition than Maple Glen.     

Isolation of transposon Tn5-induced hydrophobic mutants of a Bradyrhizohium japonicum strain with improved competitive nodulation abilities

Abstract

Hydrophobic mutants of the Bradyrhizohium japonicum strain 138NR were obtained by transposon Tn5 mutagenesis followed by replica-plating on polystyrene plates. Fifteen mutants were isolated at a frequency of 10⁶. Gel-filtration analysis of the exopolysaccharides revealed that the hydrophobic mutants produced a significantly smaller amount of low-molecular-weight polysaccharides than the parent. Four of the isolated mutants formed a larger number of nodules on soybean (Glycine max L. Merr. cv. Tamahomare) than the parent, two were superior in symbiotic nitrogen fixation, and two were symbiotically defective on soybean. Competitive nodulation abilities of the mutants were examined by inoculating them to soybean with B. japonicum strain 123ET as a competitor and determining the nodule occupancy based on the antibiotic resistance. All the mutants tested except for the symbiotically defective ones were superior in their competitive nodulation ability to the parent strain. When inoculated 24 h before the inoculation with 123ET, the mutants exhibited an increased nodule occupancy (44–93% compared to 9% by the parent).     

Symbiotic dinitrogen fixation - its dependence on plant nutrition and its ecophysiological impact

The cultivation of symbiotic legumes and the consequent small uptake of nitrate by the host plant depresses soil pH and impairs subsequent legume growth. The rate of soil pH decline depends on the soil H⁺ buffer power, climatic conditions, soil permeability, and on the kind of legume cropping. It is shown for some leguminous crop species from the temperate climate that in many cases the N₂ fixation performance of Rhizobium/Bradyrhizobium symbiosis is more dependent on the growth conditions of the host plant than on the nitrogenase potential of the bacteroid. This is true for the supply of the host plant with phosphate, potassium and water as well as for light intensity. Nitrogen deficiency of the host plant because of insufficient nitrogenase activity was observed at low soil pH where nodulation and the development of the nitrogenase activity were delayed. Nodulation and nitrogenase activity are suppressed by high levels of available nitrogen in the soil. There are indications that the nitrogenase activity is very flexible and adjusts to the demand of the host plant. The mechanism of this regulation is not yet understood.     

缺氮培养大豆Bragg结瘤突变系对CO2倍增的反应

研究了在缺氮条件下，CO2倍增对大豆（GtycinemaxL.）Bragg及其等基因突变体超结瘤大豆nts382和不结瘤大豆Nod49生长和固氮的影响。结果表明在缺氮条件下CO2倍增明显提高大生物量和根系结涵量，但对固氮酶活性的影响则随幼苗的生长而异。播种后25天取样结果显示CO2倍增条件下，Bragg和nts382的固氮比活性和单株固氮活性都显著提高，而其后3天取样的结果没有表现出增加趋势，固氮比活性在nts382反而明显降低。两种CO2浓度条件下，nts382单株固氮活性高于Bragg，但固氮比活性低于后者。两次测定结果的差异说明植物对CO2倍增的反应具有很强的时效性;同时表明，CO2倍增对植物生长和固氮的促进作用不能长期维持。这可能与生物固氮过程本身的复杂性有关。根据本研究结果推测，在未来全球环境变化、CO2倍增条件下，共生固氮植物可能在生态系统氮素平衡中起到更为重要的作用;并有可能通过育种技术改良固氮农作物，提高农作物产量。     

抗春雷霉素大豆根瘤菌突变株的诱变及其生物固氮特性的分析

根瘤菌与豆科植物共生可以固定大量的氮。根瘤菌剂接种豆科作物是一项普遍推广应用、有效的农业技术。但由于大量土著根瘤菌的存在，产生竞争障碍，降低了接种菌剂的占瘤率。大多数的土著根瘤菌对春雷霉素敏感，因此接种抗春雷霉素的高效结瘤固氮根瘤菌，并用春雷霉素处理种子，可抑制土著根瘤菌，提高接种菌剂的占瘤率，从而达到高结瘤、高固氮和提高产量的目的。本文将探讨诱变对根瘤菌抗春雷霉素突变的作用，并对获得的抗性突变株的生物固氮特性进行分析。     

共生固氮在农牧业上的作用及影响因素研究进展

共生固N是生物固N的主体部分,具有固N效率高、应用范围广等特点。叙述了主要豆科作物年固N量及固N量占豆科作物本身所吸收N的比例,阐述了豆科作物在与非豆科作物间套轮作中固定N素的转移及对非豆科作物的影响,并介绍了影响豆科作物-根瘤菌共生体共生固N效率的主要因素。开展豆科作物-根瘤菌共生体系方面的研究对农业可持续发展具有重要意义。     

缺氮培养大豆Bragg结瘤突变系对CO2倍增的反应

研究了在缺氮条件下，CO₂倍增对大豆（GtycinemaxL.）Bragg及其等基因突变体超结瘤大豆nts382和不结瘤大豆Nod49生长和固氮的影响。结果表明在缺氮条件下CO₂倍增明显提高大生物量和根系结涵量，但对固氮酶活性的影响则随幼苗的生长而异。播种后25天取样结果显示CO₂倍增条件下，Bragg和nts382的固氮比活性和单株固氮活性都显著提高，而其后3天取样的结果没有表现出增加趋势，固氮比活性在nts382反而明显降低。两种CO₂浓度条件下，nts382单株固氮活性高于Bragg，但固氮比活性低于后者。两次测定结果的差异说明植物对CO₂倍增的反应具有很强的时效性;同时表明，CO₂倍增对植物生长和固氮的促进作用不能长期维持。这可能与生物固氮过程本身的复杂性有关。根据本研究结果推测，在未来全球环境变化、CO₂倍增条件下，共生固氮植物可能在生态系统氮素平衡中起到更为重要的作用;并有可能通过育种技术改良固氮农作物，提高农作物产量。     

Hydrogenase Activity of Soybean Nodules Doubly Infected with Bradyrhizobium japonicum and B. elkanii

Rhizobitoxine (2-amino-4-(2-amino-3-hydropropoxy)-trans-but-3-erioic acid) is a phytotoxin produced by some strains of Bradyrhizobium species. Rhizobitoxine-producing strains often induce chlorosis in new leaves of soybean as a result of the synthesis of the toxin in nodules (Owens and Wright 1964; Owens et al. 1972). Some of the B. japonicum bacteroids possessing the hydrogen uptake (Hup) system are capable of ATP production by recycling H₂ evolved from nitrogenase (Evans et al. 1987). Adequate uptake hydrogenase activity in soybean bacteroids often enhances plant growth, as well as the efficiency of energy utilization during nitrogen fixation (Evans et al. 1987).     

白浆土大豆结瘤及其固氮状况

黑龙江省是我国大豆主要产区,在国营农牧场中大面积机械化生产条件下,大豆的种植面积约占耕地面积的三分之一左右。一般认为,大豆与根瘤菌的共生固氮作用所固定的氮,能提供大豆整个生育过程中对氮素需要量的二分之一到三分之二。然而,在大田生产实践中,由于土壤类型、肥力水平、气候条件、前作、品种、施肥等因素的综合影响,使大豆在田间的共生固氮效率产生了很大的差异。     

The location and distribution in soil of rhizobia under senesced annual legume pastures

Soil cores from under senesced legume swards were partitioned into (1) soils, (2) legume roots, (3) nodules and (4) debris. The viable population of Rhizobium trifolii under subterranean clover was found to be predominantly in the nodule fraction; but with R. lupini under Serradella the distribution was more even. In commercially sown stands the rhizobia often failed to migrate from the original sown rows into the soil between the rows. The relative importance of the various fractions as contributors to the “pool” of rhizobia available for nodulation of subsequent generations of host-plants is discussed.     

Evaluation of effective Myanmar Bradyrhizobium strains isolated from Myanmar soybean and effects of coinoculation with Streptomyces griseoflavus P4 for nitrogen fixation

In order to substitute the use of chemical fertilizers in legume production, there is a need for the production of rhizobial inoculants which are capable of being used as biofertilizers. To achieve this, an effective symbiotic nitrogen (N) fixation between legumes and root nodule bacteria will be essential. Evaluation of effective Myanmar Bradyrhizobium (Jordan 1982) strains isolated from Myanmar soybean (Glycine max L. Merr.) and effects of coinoculation with Streptomyces griseoflavus Krainsky 1914 P4 for N fixation were studied in pot experiments using sterilized vermiculite and Hoagland solution in the Phytotron (25°C and 70% relative humidity) with completely randomized design and three replicates. N fixation ability of soybean was evaluated by acetylene reduction activity (ARA) by gas chromatography. It was found that MAS23 showed a relatively high degree of stability and a high level of ARA per plant on both Yezin-3 and Yezin-6 soybean varieties. In the symbiotic relationship between Bradyrhizobium strains and P4 experiments, the treatments consisted of six Bradyrhizobium strains (MAS23, MAS33, MAS34, MAS43, MAS48 and USDA110) and Streptomyces griseoflavus P4 were evaluated with four Myanmar soybean varieties (Yezin-3, Yezin-6, Hinthada and Shan Sein). In the Yezin-3 soybean variety, the best treatment for ARA per plant was found in the dual inoculation of P4 and MAS34. In the Yezin-6 soybean variety, the highest nodule dry weight was found in dual inoculation of P4 with MAS34 but the highest ARA per plant was observed in the dual inoculation of P4 and MAS23. On the other hand, single inoculation of MAS43 and coinoculation of P4 with MAS48 were significantly higher in N fixation of Hinthada, and coinoculation of P4 with MAS33 was significant improvement of ARA per plant (P < 0.05) in Shan Sein soybean.     

Competitive advantage of bacteriocin and phage-producing strains of Rhizobium trifolii in mixed culture

Bacteriocinogenic and lysogenic strains of Rhizobium trifolii were grown with sensitive strains in sterile broth or peat culture. In broth culture the producing strains strongly suppressed growth of sensitive strains. In peat culture a considerable degree of suppression was also observed, the effect being stronger when the peat was “wet” than when it was “damp”. Apparently, conditions which favour growth in peat also favour dominance by bacteriocinogenic or lysogenic bacteria in cultures of mixed strains. The results are discussed in relation to preparation of mixed-strain legume inoculants and to studies of competition between strains in the field environment.