Rhizobia in tropical legumes—XIII. Biochemical basis of acid and alkali reactions

Three fast-growing rhizobia (Rhizobium meliloti isolated from Medicago saliva, R. trifolii from Trifolium subterraneum, and Rhizohium sp. from Leucaena leucocephala) and three slow-growing rhizobia R. japonicum from Glycine max, Rhizobium spp from Centrosema pubescens and Crotolaria anagyroides) were grown in defined media. The mean generation times of the fast-growing and slowgrowing strains were 3.8 h and 8.6 h respectively. Slow-growing organisms raised the initial pH of the defined medium while the fast-growing organisms lowered it. Rates of oxygen consumption tended to be higher in the slow-growing organisms.UMKL 19 (isolated from L. leucocephala) possessed all the normal reactions of fast-growing rhizobia but had a single sub-polar flagellum, similar to the three slow-growing strains studied.Certain combinations of amino acids and sugars (e.g. glutamine and galactose) induced an acidic reaction in the fast-growing organisms while the slow-growing ones changed the media to alkaline. Fast-growing organisms utilized more galactose for growth compared to slow-growing ones. Both types of organisms synthesized and released a wide range of amino acids into the medium.We suggest that pH changes produced by rhizobia growing on yeast-extract mannitol media are caused by the preferential utilization of sugars by fast-growing organisms and nitrogenous compounds by slow-growing ones.     

Influences of intercropping and nitrogen supply on flavonoid exudation in wheat roots

Abstract

Soil and hydroponic experiments were carried out to examine the influences of intercropping and nitrogen supply on flavonoid exudation in wheat roots. Both experiments comprising three cropping patterns (wheat intercropped with faba bean, monocropped wheat, and monocropped faba bean) and three N supply levels (deficient, adequate, and excessive) with three replicates in a randomized complete block design. Across two experiments, intercropping increased but N fertilization decreased flavonoids of wheat roots frequently. Intercropping variably increased secretion of naringenin from 0.5 to 1.9 folds (P?<?0.5) in wheat roots at all three N levels, but rarely increased secretion of genistein and hesperetin in wheat at the deficient N level. Intercropped wheat secreted more flavonoids than monocropped wheat at its tillering (60^th d) and flowering (95^th d) stages; after the flowering stage, however, the differences between intercropping and monocropping were not significant at any N level. Secretion of flavonoids in wheat roots decreased with increased N supply. Interspecies and N supply altered the contents and proportions of flavonoids in wheat root exudations under wheat and faba bean intercropping. These results indicate facilitative root–root interactions and provide insight into cereal promote nodule of legume in intercropping system.     

Rhizobia in tropical legumes—XIV. Ion uptake differences between fast- and slow-growing strains

We compared the uptake of nitrogen, potassium and phosphorus (as well as ¹⁴C-labelled mannitol, ³H-labelled glutamate, and ³²P-labelled phosphate) in three fast- and three slow-growing rhizobia. The fast-growing strains used were Rhizobium meliloti (isolated from Medicago sativa), R. trifolii (from Trifolium subterraneum), and Rhizobium spp from Leucaena leucocephala, while the slow-growing strains were R. japonicum (Glycine max), and two Rhizobium spp (from Centrosema pubescens and Crotolaria anagyroides). Slow-growing organisms preferentially utilized glutamate in the medium. Both fast- and slow-growing strains took up more NH⁺₄-N than NO^?₃-N on a per cell basis. In the presence of mannitol, fast-growing strains can cause either acid or alkaline reactions, an effect that is dependent only on the N-source (NH⁺₄ or NO^?₃). Uptake preferences of the fast-growing Leucaena isolate (UMKL 19) resembled those of the slow-growing rhizobia, further strengthening the argument that this organism (and others like it) may be intermediate between the normal fast- and slow-growing groups. Generally, the efficiency of uptake of N (either as NH⁺₄ or NO^?₃), P, and therefore K, was greater in the fast-growing organisms.     

Organic acid exudation by mycorrhizal Andropogon virginicus L. (broomsedge) roots in response to aluminum

Elevated aluminum (Al) availability limits plant growth on acidic soils. Although this element is found naturally in soils, acidic conditions create an environment where Al solubility increases and toxic forms of Al impact plant function. Plant resistance to Al is often attributed to organic acid exudation from plant roots and the chelation of cationic Al in the rhizosphere. The association of arbuscular mycorrhizal (AM) fungi with the roots of plants may alleviate Al toxicity by altering soil Al availability or plant exposure through the binding of Al to fungal structures or through the influence of fungi on exudation from roots. Diverse communities of AM fungi are found in soil ecosystems and research suggests that AM fungi exhibit functional diversity that may influence plant performance under varying edaphic environments. In the present study, we evaluated acidic isolates of six AM species in their responses to Al. Andropogon virginicus (broomsedge), a warm-season grass that commonly grows in a range of stressful environments including acidic soils, was used as a plant host for Acaulospora morrowiae, Glomus claroideum, Glomus clarum, Glomus etunicatum, Paraglomus brasilianum, and Scutellospora heterogama. Fungal spores were germinated and exposed to 0 or 100 μM Al on filter paper in sand culture or were grown and exposed to Al in sand culture in association with A. virginicus. Short- and long-term responses to Al were evaluated using direct measurements of fungal spore germination, hyphal elongation, and measurements of A. virginicus colonization and plant growth as a phytometer of AM function in symbio. Spore germination and hyphal elongation varied among AM species in response to Al, but patterns were not consistent with the influences of these AM species on A. virginicus under Al exposure. Exposure to Al did not influence colonization of roots, although large differences existed in colonization among fungal species. Plants colonized by G. clarum and S. heterogama exhibited the least reduction in growth when exposed to Al, produced the highest concentrations of Al-chelating organic acids, and had the lowest concentrations of free Al in their root zones. This pattern provides evidence that variation among AM fungi in Al resistance conferred to their plant hosts is associated with the exudation of Al-binding organic acids from roots and highlights the role that AM fungal diversity may play in plant performance in acidic soil environments.     

Grain legumes differ in nitrogen accumulation and remobilisation during seed filling

In grain legumes, the N requirements of growing seeds are generally greater than biological nitrogen fixation (BNF) and soil N uptake during seed filling, so that the N previously accumulated in the vegetative tissues needs to be redistributed in order to provide N to the seeds. Chickpea, field bean, pea, and white lupin were harvested at flowering and maturity to compare the relative contribution of BNF, soil N uptake, and N remobilisation to seed N. From flowering to maturity, shoot dry weight increased in all crops by approximately 50%, root did not appreciably change, and nodule decreased by 18%. The amount of plant N increased in all crops, however in field bean (17?g?m^?2) it was about twice that in chickpea, pea, and lupin. The increase was entirely due to seeds, whose N content at maturity was 26?g?m^?2 in field bean and 16?g?m^?2 in chickpea, pea, and lupin. The seed N content at maturity was higher than total N accumulation during grain filling in all crops, and endogenous N previously accumulated in vegetative parts was remobilised to fulfil the N demand of filling seeds. Nitrogen remobilisation ranged from 7?g?m^?2 in chickpea to 9?g?m^?2 in field bean, and was crucial in providing N to the seeds of chickpea, pea, and lupin (half of seed N content) but it was less important in field bean (one-third). All the vegetative organs of the plants underwent N remobilisation: shoots contributed to the N supply of seeds from 58% to 85%, roots from 11% to 37%, and nodules less than 8%. Improving grain legume yield requires either reduced N remobilisation or enhanced N supply, thus, a useful strategy is to select cultivars with high post-anthesis N₂ fixation or add mineral N at flowering.     

盐胁迫下粪产碱菌在水稻根表的定殖(英文)

粪产碱菌A1 5 0 1能在氯化钠浓度高达 4%的固体和液体介质中良好生长 ,合成IAA和固定空气中的氮素。当氯化钠浓度超过 3 %时 ,A1 5 0 1丧失运动和趋化能力。一定盐浓度胁迫下A1 5 0 1能定殖在水稻根表和根毛区和侧根伸出部位 ,表现出较高的联合固氮活性。0 5 %的氯化钠能促进A1 5 0 1在水稻根表的定殖能力。A1 5 0 1能通过侧根伸出部位侵入根内。     

Colonization of wheat seedling (Triticum aestivum) roots by Pseudomonas fluorescens and Bacillus subtilis

Summary Colonization patterns of Pseudomonas fluorescens and Bacillus subtilis on roots of wheat seedlings growing on water agar were studied qualitatively by replica printing and quantitatively by the plate count method. The results indicated a stronger colonization potential for P. fluorescens (up to 10⁷ cfu/cm root) than for B. subtilis (up to 10⁵ cfu/cm root). Although the numbers of both species were lower when inoculated together, the observed colonization patterns on the roots were comparable to those found with single inoculations. For none of these bacteria was active migration along the root surface in any direction observed, indicating that distal positions are reached mainly by a passive displacement on the root tip and elongating cells. Ecological implications of the observed phenomena are discussed.     

Colonization of wheat roots by Gaeumannomyces graminis inhibited by specific soils,microorganisms and ammonium-nitrogen

Lineal extension of Gaeumannomyces graminis var. tritici hyphae along roots of intact wheat plants growing in soils was measured. Hyphal growth rates were lower in soils treated with NH₄⁺-N than with NO₃^?-N. In a soil that is suppressive to the take-all disease, the controlling influence of NH₄⁺-N was eliminated by soil fumigation (methyl bromide), and reintroduced to fumigated soil by additions of 1% nonsterile soil. Effects of fumigation on hyphal growth were absent in a nonsuppressive soil, and in NO₃^?-treatments of the suppressive soil. When inocula of selected groups of wheat rhizoplane microflora were reintroduced into a fumigated or a soil-reinoculated soil via a root-food base, the Pseudomonas spp. consistently appeared more suppressive in NH₄⁺-N treatments than the general bacterial flora, Bacillus spp. spores, streptomycetes, and fungi.     

Antioxidant capacity of seed coat, dehulled bean, and whole black soybeans in relation to their distributions of total phenolics, phenolic acids, anthocyanins, and isoflavones

Black soybeans have been used as an excellent dietary source for disease prevention and health promotion in China for hundreds of years. However, information about the distribution of health-promoting phenolic compositions in different physical parts of black soybean and the contribution of phenolic compositions to overall antioxidant capacity is limited. To elucidate the distribution of phenolic composition and their contribution to antioxidant activities in black soybean, the total and individual phenolic profiles, and antioxidant capacities of seed coat, dehulled and whole black soybean were systematically investigated. The seed coat exhibited much higher total phenolic indexes and antioxidant activities than whole and dehulled black soybean. Dehulled black soybean possessed similar levels of total phenolic content, total flavonoid content, 2-diphenyl-1-picryhydrazyl (DPPH) radical scavenging activity, ferric reducing antioxidant power (FRAP), and oxygen radical absorbance capacity (ORAC) activities as compared to whole yellow soybean. Cyanidin-3-glucoside, petunidin-3-glucoside, and peonidin-3-glucoside were detected in the seed coat but not in dehulled black soybean and yellow soybean. Among benzoic acid detected, caffeic and chlorogenic acid were the predominant phenolic acids. Whole black soybean and dehulled black soybean exhibited similar isoflavone contents in 7- O-beta-glucosides and malonylglucosides of daidzein and genistein. The seed coat possessed significantly ( p < 0.05) lower 7- O-beta-glucosides and malonylglucosides of daidzein and genistein, acetylglycitin, and total isoflavones than whole and dehulled black soybean. The contribution of phenolics in the seed coat to the antioxidant activity of black soybean parts depends on the assay methods. When measured with the DPPH and FRAP methods, the seed coat contributed 90% of the total antioxidant capacity of black soybean. However, when measured with the ORAC method, the seed coat and dehulled portion contributed approximately equally the total antioxidant capacity of black soybeans. The information generated from this study on the distribution and content of their active components is useful for the effective use of black soybeans as an ingredient for promoting health.     

Physiological basis of iron chlorosis tolerance in rice (Oryza sativa) in relation to the root exudation capacity

Micronutrient deficiency in cultivable soil, particularly that of iron (Fe) and zinc (Zn), is a major productivity constraint in the world. Low Fe availability due to the low solubility of the oxidized ferric forms is a challenge. An experiment was, thus, executed to assess the performance of eight genetically diverse rice genotypes on Fe-sufficient (100 µM) and Fe-deficient (1 µM) nutrient solution, and their ability to recover from Fe deficiency was measured. Fe efficiency under Fe deficiency in terms of biomass production showed a significant positive correlation with the root release of phytosiderophore (PS) (R² = 0.62*). This study shows that the Fe deficiency tolerance of Pusa 33 was related to both a high release of PS by the root and an efficient translocation of Fe from the root to the shoot as the Fe–PS complex, which could be useful for improving the Fe nutrition of rice particularly under aerobic conditions.     

氯化钾对玉米根系糖和酚酸分泌的影响及其与茎腐病菌生长的关系

通过溶液培养试验,研究了氯化钾（KCl）对不同抗性品种玉米根系糖和酚酸分泌量的影响,以及在不同浓度糖和酚酸的培养基上禾谷镰刀菌（Fusarium graminearum Schwabe）的生长状况,探讨KCl抑制玉米茎腐病发生过程中,根系分泌物中糖和酚酸所发挥的作用。结果表明,感病品种吉单327的总糖、还原糖和其蔗糖分泌量均高于抗病品种吉单180;正常供钾条件下,总糖、还原糖和蔗糖的分泌量均表现出不同程度的下降,且还原糖下降幅度最大。阿魏酸和绿原酸是玉米根系分泌物中主要的两种酚酸组分。抗病品种吉单180分泌的阿魏酸量明显高于感病品种吉单327,而绿原酸分泌量低于吉单327。正常供钾时,阿魏酸分泌量明显增加,但绿原酸分泌量则有所减少。一定浓度的蔗糖和葡萄糖均能显著促进F. graminearum的生长,且还原糖（葡萄糖）的促进效果较蔗糖更明显;阿魏酸和绿原酸均能抑制F. graminearum的生长,但阿魏酸的抑制效果远远高于绿原酸。KCl 可明显减少不同抗性品种玉米根系的还原糖分泌量,增加阿魏酸的分泌,从而一定程度上抑制了病原菌F. graminearum在根际的快速生长。     

氮磷比对大豆生物学性状和根系分泌有机酸量的影响

为阐明生长介质中的氮磷条件对大豆生长和根系分泌有机酸的影响, 采用沙培培养方法, 设计3 种不同的氮磷配比, 分析不同氮磷条件下大豆生物量积累和结瘤情况, 利用高效液相色谱分析根系分泌的有机酸, 探讨根系分泌有机酸的变化情况。结果表明, 在大豆苗期, 适当减少磷肥施用量不影响生物量积累, 但需要充足的氮才能保证结瘤性状。低磷促进大豆根系分泌有机酸, 高氮对低磷处理有机酸分泌有促进作用, 说明磷和氮对大豆根系分泌有机酸存在负交互作用。     

Effect of seed size and phosphorus fertilization on growth of selected legumes

Abstract

A screenhouse experiment was conducted to assess the effects of seed size and phosphorus (P) fertilization on growth of 12 herbaceous and shrub legumes [Arachis hypogaea, Cajanus cajan, Centrosema pascuorum, Centrosema brasilianum, Crotolaria ochroleuca, Glycine max, Lablab purpureus, Mucuna cochinchinensis, Mucuna pruriens var. utilis (black seed), Mucunapruriens (white seed), Pueraria phaseoloides and Vigna unguiculata] grown on a P‐deficient Plinthustalf. Species with large seed size showed higher biomass accumulation, nodulation and higher plant nutrient element content. However, response to P fertilization was higher in small size seeded species. Plant top dry weight was highly correlated with seed weight and seed nutrient element contents. Although P application significantly increased nitrogen (N), P, potassium (K), calcium (Ca), and magnesium (Mg) contents of plant tops, high rate of P fertilization appeared not to be beneficial to the early growth of the species tested on this P‐deficient soil. The use of large seed size with high seed nutrient element contents is highly recommended for fast establish ment of fallow legume species intended for use in production systems on P‐deficient soils.     

Studies on the change of respiratory system in roots in relation to the growth of plants

It has been observed that in nitrogen starved rice plants, the length of the root increased and became voluminous. The colour of the root appeared pale. Both the oxygen uptake and the oxidative power of the roots decreased during the vegetative stage of the plant (1, 2, 3). Under conditions of K deficiency, the length of the roots became shorter and the colour was light grayish black. These roots were very susceptible to damage (3). K deficient roots showed a low oxidative power (3, 4), while the uptake of oxygen was high during the vegetative stage of growth of the plant.     

Studies on the change of the respiratory system in roots in relation to the growth of plants

Extensive studies on respiration and nutrient absorption by plant and algal tissues have been carried out in recent years. These studies strongly support the evidence that cellular respiration and the assimilation of inorganic nitrogen are closely interdependent metabolic processes (1–5).     

Simulation of the effect of citrate exudation from roots on the plant availability of phosphate adsorbed on goethite

Rhizosphere processes strongly influence the availability of phosphorus (P) to plants. Organic ligands that are exuded from the root surface mobilize phosphorus by dissolution of P minerals or by desorption of adsorbed phosphate. We developed a mechanistic model to study the mobilization of phosphate sorbed on goethite by the exudation of citrate and consequent uptake of phosphate by the root. The use of a model allows the effects of the organic anion and pH on P desorption to be separated. The model is also used to predict concentration profiles developing around the root for phosphate, citrate (with or without accounting for degradation) and pH, providing insight into the processes that occur in the rhizosphere. Results of model calculations show that with larger rates of citrate exudation, greater P availability is predicted. Exudation at a rate of 0.5 μmol citrate m^–1 root day^–1, which is in the range found for P-deficient plants, increased P availability almost 2-fold at fairly large phosphate loading of goethite (1.9 μmol m^–2) and almost 30-fold at small phosphate loading (1.3 μmol m^–2). Competitive adsorption causes a much greater relative increase in the phosphate concentration in solution at small than at large phosphate loading, which explains this result. Simultaneous acidification of the rhizosphere results in a smaller P mobilization than at a fixed pH of 5, as a result of the pH dependence of phosphate adsorption in the presence of citrate. Sorption of citrate increases its persistence against microbial decay, and hence has a positive effect on the mobilization of adsorbed phosphate.     

Medium-term seed storage of 50 genera of forage legumes and evidence-based genebank monitoring intervals

Genebanks maintaining seeds for long-term genetic resources conservation monitor seed lots to detect early loss in viability. Monitoring is costly and depletes valuable seed. Three decades of genebank seed germination test results of diverse forage species from 50 legume genera in the International Livestock Research Institute’s medium-term store (circa 8 °C with 5% moisture content) were analysed to determine whether advice on seed monitoring intervals could be derived. Cumulative normal distributions were fitted by probit analysis for each seed lot and compared within each genus. Six patterns of within-genus variation were identified: no detectable trend in germination test results during storage (4 genera); detectable trends, but variable (positive to negative) amongst lots (5); consistent slope of loss in viability amongst lots (17); consistent slope of increase in ability to germinate amongst lots (21); common loss in viability amongst lots (2); common increase in ability to germinate amongst lots (1). Seed lot monitoring intervals for the medium-term store were derived for each of 19 genera with consistent loss in viability across seed lots: three genera provided comparatively rapid deterioration, five met the general expectations for a medium-term store (2–10 years’ maintenance of high viability), whilst 11 provided much better survival. Moreover, 26 further genera provided no evidence as yet of seed deterioration; of these, 22 improved in ability to germinate during storage indicating confounding of hardseededness with viability in germination tests.     

豌豆根瘤菌的盐碱适应性鉴定

通过模拟试验,研究了筛选自甘肃武威、临夏、庆阳、定西的13株豌豆族根瘤菌对酸碱和盐分的适应性.结果表明:筛选菌株的盐碱忍耐能力凶菌株而异,菌株D2的耐盐能力最强,其OD600nm在2.5%、3.5%、4.5%的NaCl浓度下,比与其差异显著且紧邻的菌株的OD600nm分别高77%、191%和112%,是最低OD600nm值的30.32倍,20.76倍和67.68倍;Q3对3.5%和4.5%NaCl的忍耐能力最低.菌株D2的耐酸能力最强,pH=4时,其OD600nm显著大于其它12株菌株,是最低值的20倍,是其它12株平均值的6.9倍;菌株W1的OD600nm在pH=10及pH=11时均为13株菌株的最大值,比与其差异显著的菌株的平均值分别高41%和135%,W1的耐碱能力最强.     

Root exudation,organic acids,and element distribution in roots of Norway spruce seedlings treated with aluminum in hydroponics

Seedlings of Norway spruce (Picea abies [L.] Karst.), which had been grown under sterile conditions for three months, were treated for one week in a hydroculture system with either 500 μM AlCl₃ or 750 μM CaCl₂ solutions at pH 4. Organic acids were determined in hot‐water extracts of ground root tissue. Oxalate (3.3—6.6 μmol (g root dry weight)^—1) was most abundant. Malate, citrate, formate, acetate, and lactate concentrations ranged between 1—2 μmol (g root dry weight)^—1. Organic substances and phosphate found in the treatment solutions at the end of the experimental period were considered to be root exudates. Total root exudation within a 2‐day period ranged from 20—40 μmol C (g root weight)^—1. In root exudates, organic acids, and total carbohydrates, total amino acids, and total phenolic substances were quantified. Citrate and malate, although present in hot‐water extracts of root tissue, were not detected in root exudates. Phosphate was released from Ca‐treated plants. In Al treatments, there was indication of Al phosphate precipitation at the root surface. Oxalate and phenolics present in the exudates of Norway spruce seedlings are ligands that can form stable complexes with Al. However, concentrations of these substances in the treatment solutions were at micromolar levels. Their importance for the protection of the sensitive root apex under natural conditions is discussed.     

Promotion of the seed germination and seedling growth of cabbage by root-colonizing fungi isolated from oat and Eucalyptus roots

(Jpn. J. Soil Sci.Plant Nutr., 77, 265–272, 2006)

We isolated fungi colonizing oat (Avena sativa L.) and eucalyptus (Eucalyptus camaldulensis Dehnh) roots to select the strains that promote the seed germination and seedling growth of cabbage. Twenty-nine strains were inoculated into the soil which grows cabbage seedlings. This inoculation test was repeated three times. From the results of the test, we found that MT0011 and EU0013 strains promoted seed germination. In the first inoculation test, seven strains increased the dry weight of cabbage seedling significantly compared with the control. In the second test with the strains that promoted cabbage growth, MT0008 and EU0013 strains significantly increased the dry weight of seedlings. In the third test, only EU0013 significantly increased seedling growth. The ITS region for DNA (ITS1-5·8SrDNA-ITS2) of 28 isolated strains were sequenced to examine the molecular phylogeny. Isolates were classified into two divisions, Zygomycota and Ascomycota. Nine strains showed the highest similarities with same accession number AF504832 (Uncultured fungus clone D50). MT0011 was clustered with fungi belonging to Drechela sp. from the sequence data on the rDNA ITS region. EU0013 was identified as Penicillium sp. from the morphological feature conidiophore and the sequence of the ITS region.