河西走廊豆科植物结瘤固氮特性初步研究

对甘肃河西走产野生豆科植物结瘤及固氮状况进行了调查。栽培豆科植物均能自然结瘤,野生豆科植物结瘤率仅５０％。该地区豆科植物根瘤大多数圆形、棒状或指状,形状较为规则。栽培植物根瘤多为粉红色,而野生揿为白色、黄色或棕色。ＡＲＡ测定表明,２５．６％根瘤为无效根瘤。不同种不同地区根瘤固氮活性相差较大,一般活力都较低,乙炔还原活性小１μｍｏｌＣ２Ｈ４／ｇＦＷ．ｈ的占检测样品数的２９．９％,最高者信为２８．１９     

新疆干旱地区根瘤菌资源研究I．根瘤菌种类及其共生固氮作用

从新疆不同生态条件下生长的31属109种豆科植物根瘤分离获得373株根瘤菌,其中88株是从未报道过结瘤情况的39种豆科植物根瘤分离获得。它们与豆科寄主植物共生结瘤,95％以上为有效根瘤。不同种的根瘤固氮活性差异较大,黄芪属根瘤固氮活性较高,最高者为大豆根瘤固氮活性的42倍。 20属37种豆科植物根瘤中97％有吸氢活性。根瘤固氮活性、吸氢活性均与寄主植物生长发育期有相关性。     

新疆干旱区豆科植物结瘤的固氮特性

调查了新疆干旱区72种豆科植物的结瘤固氮活性,其中33种尚未见报道。这些植物所结根瘤在外形上多数不规则,以皮层厚和白色、棕色者居多,与非干旱区的根瘤形态显著不同。根瘤固氮活力相差较大,但比一般豆科植物根瘤活性高,最高者可达当地大豆根瘤的42倍。根瘤活性与宿主的抗逆境能力有关。此外,从11种豆科植物根瘤观察到10种具有吸氢活性。对干旱区豆科共生固氮生理生态的特性进行了讨论。     

高氮抑制豆科植物结瘤固氮机制研究进展

豆科植物通过与根瘤菌共生,形成根瘤并进行生物固氮。豆科植物的结瘤固氮作用在农业上具有减肥增效、改良土壤等重大意义。然而,高氮会抑制豆科植物结瘤固氮,形成"氮阻遏"效应。着重论述了高氮抑制豆科植物结瘤的分子机制,包括氮素通过结瘤自调控AON(Autoregulation of nodulation)信号、NLP(NIN-like protein)转录因子、植物激素信号等途径抑制根瘤数目和发育的最新进展;并探讨了高氮抑制根瘤固氮活性的假说及争议,包括亚硝酸盐毒性和碳饥饿等,以期为提高豆科植物应对"氮阻遏"效应提供理论基础。     

气候因子对银合欢共生固氮的影响

本文研究气候因子对银合欢共生固氮的影响。银合欢结瘤固氮有明显的季节性变化,夏秋结瘤多,固氮酶活性高,冬天及早春结瘤少,活性低。银合欢结瘤固氮的适宜温度为25-30℃,低于10℃或高于30℃时活性显著下降,处于0℃或45℃时活性极低,逆境温度处理时间越长活性越低。温度相近的条件下,土壤湿度对银合欢结瘤固氮的影响尤其突出,水分过多或干旱都严重影响结瘤固氮。遮光条件下银合欢根瘤固氮酶活性显著降低,晴天取样的根瘤固氮酶活性比阴天的高。银合欢根瘤固氮酶活性在一天中出现两个峰,第一个峰在11时,第二个峰在17-20时。离体根瘤的固氮酶活性随离体时间的延长而降低,表明植株光合作用产物对根瘤的供应及其在根瘤中的贮备对固氮酶活性有显著影响。     

台湾相思结瘤固氮与吸氢酶活性研究

台湾相思的根系具有多年生的根瘤,根瘤初发生时球状,以后发育成分叉瘤和扇状瘤。根瘤固氮活性因苗龄、成熟度不同而有明显差异。环境条件影响结瘤及固氮活性。15℃时结瘤受到明显抑制,固氮作用最适温度条件是25～30℃。光照不足降低根瘤固氮活性。短期轻度干旱不影响根瘤固氮活性,但持续干旱使固氮活性明显下降。pH4.5～8.5条件能正常结瘤,pH5.5时结瘤最好。根瘤固氮作用时不释放H_2,具有较高的吸氢酶活性,在固氮反应系统中加入5％的H_2,能提高根瘤固氮活性。     

几种非豆科植物根瘤内生菌侵染特征的研究

自不同科、属、种的非豆科植物根瘤分离内生菌,对其寄主植物进行了交叉侵染,结果表明,这些Frankia菌对不同寄主的侵染没有明显的专一性,供试菌可以进行跨越科、属、种的侵染,但有的菌株对于某些植物的侵染,可能存在一些特殊情况,相同菌株对不同植物的侵染能力,以及不同菌株对同一寄主的侵染能力是有差异的。从同一种植物根瘤中分离的不同菌株,侵染能力也有高低之分,供试菌随寄主植物的改变,侵染能力及所建立的共生系统固氮活性有所降低,侵染原寄主植物所形成的根瘤固氮活性较高的菌株,在改变寄主后所形成的根瘤固氮活性也比较高,在一定条件下,寄主植物的结瘤量与根瘤固氮活性呈正相关,而侵染不同寄主后,根瘤中菌体孢子的表面结构也发生了一定变化。     

几种非豆科植物根瘤内生菌侵染特征的研究

自不同科、属、种的非豆科植物根瘤分离内生菌,对其寄主植物进行了交叉侵染,结果表明,这些Frankia菌对不同寄主的侵染没有明显的专一性,供试菌可以进行跨越科、属、种的侵染,但有的菌株对于某些植物的侵染,可能存在一些特殊情况,相同菌株对不同植物的侵染能力,以及不同菌株对同一寄主的侵染能力是有差异的。从同一种植物根瘤中分离的不同菌株,侵染能力也有高低之分,供试菌随寄主植物的改变,侵染能力及所建立的共生系统固氮活性有所降低,侵染原寄主植物所形成的根瘤固氮活性较高的菌株,在改变寄主后所形成的根瘤固氮活性也比较高,在一定条件下,寄主植物的结瘤量与根瘤固氮活性呈正相关,而侵染不同寄主后,根瘤中菌体孢子的表面结构也发生了一定变化。     

大豆品种资源携带的无效结瘤Rj2基因的研究

通过黑龙江省野生、半野生和栽培大豆与美国大豆根瘤菌VSDA122结瘤反应试验,本文首次报导了无效结瘤R_i2基因广泛存在中国东北大豆中,无效结瘤率为64%;同时导致大豆植株干鲜重及根瘤固氮酶活性下降,以及产生缺绿病.     

陕西及甘,宁部分地区豆科植物根瘤菌资源调查

通过对陕西全省及甘、宁部分地区豆科植物瘤菌资源进行调查,共采集到４４属１０９种豆科植物根瘤３８７份,其中蝶形花亚科中有９４．６％的植株能结瘤固氮,槐属和香槐属中的一些种及云实亚科中的云实和紫荆未见结瘤。对部分菌株的固氮酶活性进行了测定,自然瘤的固氮酶活性比回接瘤的固氮酶活性低。     

盐生野大豆的异黄酮积累及其生态学意义

以自然生长在盐碱地上的野大豆(Glycine soja)和不耐盐的栽培大豆(G. max)为材料,测定了它们在不同盐度条件下叶片、根部和种子的异黄酮含量,并测定了它们叶片的L-苯丙氨酸含量和苯丙氨酸裂解酶(PAL)活性,还测定了它们根部的结瘤量和固氮酶活性。通过两者比较,分析了它们的大豆异黄酮代谢和盐渍环境的关系。结果表明:盐渍处理不抑制盐生野大豆PAL酶的活性,其大豆异黄酮大量积累;相反,盐渍处理明显抑制栽培大豆PAL酶活性,其大豆异黄酮含量减少,而大豆异黄酮合成前体L-苯丙氨酸积累。结果还显示:在盐渍条件下,盐生野大豆根部异黄酮积累的同时,其根瘤结瘤量较多,且固氮酶活性也较高;而栽培大豆随着其根部异黄酮的减少,其根瘤结瘤量大大减少,且固氮活性大大下降。野大豆和栽培大豆的这些差别说明:盐生野大豆积累大豆异黄酮有其生态学意义,这很可能是野大豆通过异黄酮次生代谢途径适应盐渍环境的一种重要机制。     

Nitrogen fixation in several grain legume species with contrasting sensitivities to copper nutrition

The nitrogen fixation response to copper nutrition in faba bean, yellow lupin and soybean was studied. Copper nutrition significantly increased the pod yields of all tested grain legumes but faba bean gave the greatest Cu-use efficiency for pod and grain production. The accumulation of dry matter in vegetative parts, nodules, N and leghemoglobin concentration in nodules and nitrogen accumulation in the whole plants were increased by copper supply in faba bean and yellow lupin in contrast with soybean. Cu nutrition significantly increased the Cu concentrations in nodules of all cultivated plants. The differential sensitivity of N₂ fixation in tested grain legume species to copper nutrition could be connected with the level of phenols in nodules and depended on both the host plants and strains of rhizobia, which differ in their ability to produce catechol-like siderophores. Copper requirements by symbiotic N₂ fixation could also depend on the nature of phenols in nodules (presence of o-dihydroxyphenols or number of hydroxyls in molecule).     

Gene centres,a source for genetic variants in symbiotic nitrogen fixation: Host-induced ineffectivity inPisum sativum ecotype fulvum

Summary Pisum sativum ecotype fulvum forms ineffective nodules with Rhizobium strains, isolated from effective nodules of the cultivated pea in Europe. Rhizobium strains isolated from nodules of fulvum peas in Israel are fully effective on this host plant, but in association with the cultivated pea they induce nodules of poor N₂-fixing activity. The distribution of these fulvum-specific Rhizobium strains is restricted to regions where the fulvum pea occurs naturally. Rhizobium strains from other geographical regions induce mainly ineffective, or partially effective nodules on fulvum plants.A wide genetic variation, with regard to symbiotic response to a standard set of Rhizobium strains, was demonstrated in the fulvum plants collected in Israel. Based on variation in N₂-fixation three groups of plants can be distinguished. These plants offer the possibility for the study of host-genetic control on symbiotic nitrogen fixation.     

Photorespiratory metabolism and nodule function: behavior of Lotus japonicus mutants deficient in plastid glutamine synthetase

Two photorespiratory mutants of Lotus japonicus deficient in plastid glutamine synthetase (GS(2)) were examined for their capacity to establish symbiotic association with Mesorhizobium loti bacteria. Biosynthetic glutamine synthetase (GS) activity was reduced by around 40% in crude nodule extracts from mutant plants as compared with the wild type (WT). Western blot analysis further confirmed the lack of GS(2) polypeptide in mutant nodules. The decrease in GS activity affected the nodular carbon metabolism under high CO(2) (suppressed photorespiration) conditions, although mutant plants were able to form nodules and fix atmospheric nitrogen. However, when WT and mutant plants were transferred to an ordinary air atmosphere (photorespiratory active conditions) the nodulation process and nitrogen fixation were substantially affected, particularly in mutant plants. The number and fresh weight of mutant nodules as well as acetylene reduction activity showed a strong inhibition compared with WT plants. Optical microscopy studies from mutant plant nodules revealed the anticipated senescence phenotype linked to an important reduction in starch and sucrose levels. These results show that, in Lotus japonicus, photorespiration and, particularly, GS(2) deficiency result in profound limitations in carbon metabolism that affect the nodulation process and nitrogen fixation.     

Nitrogen Fixation in Wild Legumes

Isolations of the rhizobia from nodules of field plants of Ulexeuropaeus and Medicago lupulina have been examined for effectivenessin fixation of nitrogen by inoculating them into host plantsgrowing in a medium free of combined nitrogen. The results indicatethat vigorous fixation may be presumed to occur in associationwith field plants of the Ulex species, but that fixation inMedicago is probably decreased by the presence of ineffectiverhizobia in some nodules. The general importance of fixationin wild legumes is discussed, and it is pointed out that dueattention must be paid to non-legume genera with root nodules.These genera are locally abundant, and in former periods werestill more prominent in some regions of the world.     

Non-leguminous seed plants in Southern Africa which fix nitrogen symbiotically

Summary Of the various groups of non-leguminous seed plants that have been implicated by workers in other regions in symbiotic nitrogen fixation, though on evidence varying in quality, the indigenous flora of southern Africa includes 1 species of Stangeria, 26 of Encephalartos, 4 of Podocarpus, 9 of Myrica, 1 of Psychotria, 41 of Pavetta, 19 of Dioscorea and 1 of Gunnera. It is probable that all the indigenous cycads bear the coralloid, alga-infested roots found on cycads of other regions; evidence that nitrogen fixation occurs in these structures is presented for two of the species. The indigenous podocarps bear the numerous, nodular structures on their roots found in other species, and tests with one species indicated that a very small fixation of nitrogen might be associated with the nodules. All the 9 species of Myrica were found to bear root nodules which are shown to fix nitrogen vigorously. All the indigenous species of Pavetta that were examined bear leaf nodules; tests on two species indicated that nitrogen fixation is associated with the nodulated leaves, but only to a slight extent, so that the plants are unable to make appreciable growth in a nitrogen-free rooting medium. In testing for the occurrence of nitrogen fixation the N¹⁵, acetylene-reduction and long-term growth methods were used.     

Studies on the Relationship Between Photosynthesis and Nitrogen Fixation in the Symbiotic System of Soybean and Nodule Bacteria( Rhizobium)

The relationship between photosynthesis of soybean and nitrogen fixation of the nodules by symbiotic Rhizobium was studied. The contents of total nitrogen and chlorophyll, the net photosynthetic rate and seed yield of soybean were much higher in either hydroponically cultivated or field-grown plants inoculated with Rhizobium B16–11C (or Clark nodulating strain) than in control without inoculation (or Clark non-nodulating strain). These results show that the symbiotic nitrogen fixation has a beneficial effect on photosynthesis. However, the effect was indirect and slow so that there was no change in the net photosynthetic rate of the soybean leaves until three clays after removing nodules from the soybean roots. On the other hand, decreasing the photosynthate supply to nodule by shade, defoliation or shoot removal of the soybean, the nodule activity declined significantly. It seems that the supply of photosynthate to root nodule is a limiting factor for symbiotic nitrogen fixation. However, the diurnal variation of the nodule activity could not be explained by change neither in the contents of sucrose and starch of the root nodules nor in the ambient temperature. The factor controlling the diurnal variation deserves further study.     

Nitrogen fixation and carbon metabolism in legume nodules

A large amount of energy is utilized by legume nodules for the fixation of nitrogen and assimilation of fixed nitrogen (ammonia) into organic compounds. The source of energy is provided in the form of photosynthates by the host plant. Phosphoenol pyruvate carboxylase (PEPC) enzyme, which is responsible for carbon dioxide fixation in C4 and crassulacean acid metabolism plants, has also been found to play an important role in carbon metabolism in legume root nodule. PEPC-mediated CO2 fixation in nodules results in the synthesis of C4 dicarboxylic acids, viz. aspartate, malate, fumarate etc. which can be transported into bacteroids with the intervention of dicarboxylate transporter (DCT) protein. PEPC has been purified from the root nodules of few legume species. Information on the relationship between nitrogen fixation and carbon metabolism through PEPC in leguminous plants is scanty and incoherent. This review summarizes the various aspects of carbon and nitrogen metabolism in legume root nodules.     

Isolation and symbiotic characterization of aromatic amino acid auxotrophs of Sinorhizobium meliloti

Ten aromatic amino acid auxotrophs of Sinorhizobium meliloti (previously called Rhizobium meliloti) Rmd201 were generated by random mutagenesis with transposon Tn5 and their symbiotic properties were studied. Normal symbiotic activity, as indicated by morphological features, was observed in the tryptophan synthase mutants and the lone tyrosine mutant. The trpE and aro mutants fixed trace amounts of nitrogen whereas the phe mutant was completely ineffective in nitrogen fixation. Histology of the nodules induced by trpE and aro mutants exhibited striking similarities. Each of these nodules contained an extended infection zone and a poorly developed nitrogen fixation zone. Transmission electron microscopic studies revealed that the bacteroids in the extended infection zone of these nodules did not show maturation tendency. A leaky mutant, which has a mutation in trpC, trpD, or trpF gene, was partially effective in nitrogen fixation. The histology of the nodules induced by this strain was like that of the nodules induced by the parental strain but the inoculated plants were stunted. These studies demonstrated the involvement of anthranilic acid and at least one more intermediate of tryptophan biosynthetic pathway in bacteroidal maturation and nitrogen fixation in S. meliloti. The alfalfa plant host seems to provide tryptophan and tyrosine but not phenylalanine to bacteroids in nodules.