接种不同根瘤菌对紫花苜蓿固氮效能及生物量的影响

为了在晋北生态区给主要推广品种筛选适宜的高效根瘤菌,使紫花苜蓿-根瘤菌共生体充分发挥共生固氮作用,建立高效、高产、优质紫花苜蓿生产体系。将中国农业科学院菌种保藏中心筛选出的9株紫花苜蓿根瘤菌作为试验材料,以山西大同地区的土壤为基质,结合温室盆栽和田间试验,比较供试材料的结瘤数、固氮酶活性、株高、生物量等指标的变化情况。筛选出适合供试材料巨能耐盐紫花苜蓿品种的4株高效根瘤菌CCBAU30015、CCBAU NMC024-1、HBU07004和CCBAU30198,并将这4株菌进行田间接种效果验证。结果表明,接种CCBAU30015、CCBAU NMC024-1、HBU07004和CCBAU30198后,全年干草产量分别比对照增加18.7%,15.8%,11.3%和13.3%,单位面积蛋白质产量分别比对照增加10.9%,10.2%,9.7%和10.1%,其中,CCBAU30015表现最为突出,在各个指标上与对照间差异均显著(P0.05)。说明通过土壤筛选的高效根瘤菌可以应用到实际生产中。     

一株固氮芽孢杆菌的分离与鉴定

为从土壤中分离得到高固氮效能的芽孢杆菌,为固氮微生物肥料的开发提供优良的菌株,从小麦的根际土壤中分离得到能在无氮培养基上生长的芽孢杆菌株,然后采用Touch-Up PCR（上升PCR）的方法从菌株中扩增固氮酶的nifH结构基因和用全氮比色法测定固氮效能的方法,复筛出具有nifH结构基因同时具有高固氮酶活的菌株,进一步从形态、生理生化、16S rDNA序列分析等方面对其进行鉴定。结果表明从小麦的根际土壤中初筛得到能在无氮培养基上生长的5株芽孢杆菌,然后复筛得到1株抗逆性强、具有nifH结构基因同时具有高固氮效能的菌株C₂,经鉴定为巨大芽孢杆菌(Bacillus megaterium)。     

Effect of Deep Placement of N Fertilizers and Different Inoculation Methods of Bradyrhizobia on Growth,N2 Fixation Activity and N Absorption Rate of Field‐grown Soybean Plants

The effects of deep placement (supplied at 20 cm depth from soil surface below plants) of 100 kg N ha^?1 of N fertilizers, urea, coated urea or calcium cyanamide (lime nitrogen) on the growth, nitrogen fixation activity, nitrogen absorption rate and seed yield of soybean (Glycine max L. Merr.) plants were examined by comparing them with control plots without deep placement of N fertilizer in sandy dune field. In addition, three different inoculation methods of bradyrhizobia were used for each N treatment: (1) transplantation of 10‐day‐old seedling in a paper pot with vermiculite inoculated with Bradyrhizobium japonicum USDA110, (2) direct transplantation of inoculated 10‐day‐old seedlings, and (3) transplantation of 10‐day‐old seedlings in a non‐inoculated paper pot. The deep placement of N fertilizers, especially calcium cyanamide and coated urea, markedly increased the growth and total N accumulation in shoot, roots and nodules, which resulted in an increase in seed yield. Daily N₂ fixation activity and N absorption rate were estimated by relative abundance of ureide‐N analysed from the concentration of N constituents (ureide‐N, amide‐N and nitrate‐N) in root bleeding xylem sap and increase in total N accumulation in whole plants at R1, R3, R5 and R7 stages. The total amount of N₂ fixation was about 50 % higher in the plants with calcium cyanamide and coated urea deep placements compared with control plants. Deep placement of slow release fertilizers kept nodule dry weight higher in the maturing stage of seed, possibly through abundant supply of photoassimilate to the nodules by supporting leaf area and activity until late reproductive stages. The results indicate that deep placement of calcium cyanamide or coated urea enhances N₂ fixation activity, which ultimately increases the seed yield. The promotive effect was observed with the seedlings transplanted in paper pot with inoculum of bradyrhizobia within any treatments, although nodulation by indigenous rhizobia was observed in the plants transplanted with non‐inoculated paper pot.     

Variation in Nitrate Tolerance of Nitrogen Fixation in the Pea/Rhizobium Symbiosis

The short-term effects of nitrate on symbiotic nitrogen fixation were studied in six cultivars of pea (Pisum sativum L.) using two strains of Rhizobium leguminosarum. Plants grown symbiotically for three weeks, in order to eliminate the effect of NO₃ on nodule formation, were exposed to nil, 5 or 15 mM NO₃ for a period of seven days. Nodule mass, nitrogenase activity by acetylene reduction and the NO₂ and NO₃ concentrations in nodules were determined. Exposure to nitrate reduced the total acetylene reduction activity (ARA) in all cultivars, and a significant cultivar × nitrate treatment interaction was found. Nitrate influenced the components of N₂ fixation, nodule mass and specific ARA (ARA/mg nodule), differently within and among cultivars. Nodule mass was reduced in all cultivars, but a variation among cultivars in the degree of reduction was found. Nitrate also reduced the specific ARA to different extents in the cultivars- The specific ARA of nodules from NO₃-treated plants, expressed as per cent of the specific ARA of nodules from plants grown without NOT₃ was negatively correlated with the concentration of NO₃ in nodules. No relationship was found between the reduction of N₂ fixation and the nitrite concentration in nodules. The results indicate that it is possible to improve the nitrate tolerance of symbiotic N₂ fixation of the established pea/Rhizobium symbiosis by selection of suitable host plants.     

Responses of Root Nodule Formation and Nitrogen Fixation Activity to Nitrate in a Split-Root System in Peanut (Arachis hypogaea L.)

Peanut is an important constituent of crop rotation systems with various vegetable crops, which need large amounts of inorganic nitrogen. Nitrate inhibits nodule formation and nitrogen fixation of the peanut plants in these cropping systems. To elucidate this nitrate-induced inhibition, an experimental method using a split-root system was designed and the inhibitory level of nitrate (14 m M ) was applied to half of the roots for 5 or 30 days in pot experiments. In the application of nitrate for 5 days, the dry weight, the nitrogen content, and both the number and the fresh weight of nodules for each half of the roots were unaffected by the nitrate applied to the other half. However, the nitrate induced significantly lower nitrogenase activity in the applied half. In the application for 30 days, nitrate induced a significantly higher nitrogen content in the applied half of the roots, and the inhibitory effect on nodulation and nodule development was observed not only in the applied half but also in the other half. Thus, the inhibitory effect of long-term nitrate application on the nodulation and nitrogenase activity of peanut may be systemic, but that of short-term nitrate application, in which external nitrate may regulate nitrogen fixation, may not be systemic.     

接种中华根瘤菌SD101和避光对紫花苜蓿固氮和光合作用的影响

为了研究光合和氮素互作对紫花苜蓿自身生长和光合作用的影响,以紫花苜蓿品种WL319为试验材料,通过接种中华根瘤菌SD101和避光处理相结合,测定接种中华根瘤菌SD101紫花苜蓿在非避光、避光和复光条件下的生长情况和光合指标。研究结果表明,非避光条件下,接种中华根瘤菌SD101能够显著提高紫花苜蓿的地上生物量、叶片中叶绿素含量和紫花苜蓿的光合能力;避光条件下,受气孔导度和非气孔等因素的影响,中华根瘤菌SD101不能发挥其与紫花苜蓿共生固氮的优越性,光合速率比不接种中华根瘤菌SD101对照低。恢复光照后,接种中华根瘤菌SD101的紫花苜蓿光合作用恢复程度没有不接菌对照好。由此可见,紫花苜蓿与中华根瘤菌SD101的共生固氮作用需要在适宜的光照下才能促进紫花苜蓿生长和光合作用。     

Effects of Dicyandiamide on Potato Performance

A nitrification inhibitor which blocks the conversion of ammonium to nitrate may reduce the risk of leaching of nitrate during the growing season. In field experiments in 1988—1990 a nitrogen fertilizer enriched with the nitrification inhibitor dicyandiamid (Alzon) was compared with a standard calcium-ammonium-nitrate fertilizer (CAN), using rates of N supply up to 360 kg ha⁻¹. There were no significant differences between CAN and Alzon with respect to: stem length, apical lateral branchmg, fresh and dry weights of haulms and tubers at maturity, and nitrogen uptake. Alzon-treated plants showed lower concentrations of nitrate in stems and leaves than CAN-treated plants.     

Confirmation of a 4BL/5RL Wheat-Rye Chromosome Translocation Line in the Wheat Cultivar ''Viking'' Showing High Copper Efficiency

Populations of 64 taxa of Trifolium were assessed for nitrogen fixation and dry matter production with and without added mineral nitrogen. The number of rhizobia per gram of compost capable of inducing nodulation was tested in 7 species and ranged from 3 to 114, showing interspecific variation for nodulation with indigenous rhizobia. In a sample of 8 species, nitrogen fixation was assessed by acetylene reduction and nodule scoring. There was a good correlation (84 %, P ≤ 0.01) between the two methods, and therefore nodule score was used as a measure of nitrogen fixation in the main assessment. There were large differences between populations in terms of shoot and root dry weight, the shoot: root ratio and nodule score. Added nitrate had a significant effect on all characters, increasing shoot dry weight and the shoot: root ratio and decreasing root dry weight and the nodule score. Results showed the importance of assessing legume germplasm with and without added nitrate in locations where suitable rhizobia for the nodulation of exotic species might not be present.     

不同供氮形态对烤烟烘烤特性的影响

为探明不同供氮形态对烤烟烘烤特性的影响。采用大田试验方法,以烤烟品种K326为试验材料,设置5个不同供氮形态的处理,即N1(100%硝态氮)、N2(30%氨态氮+70%硝态氮)、N3(50%氨态氮+50%硝态氮)、N4(70%氨态氮+30%硝态氮)、N5(100%氨态氮),研究烘烤过程中不同处理的烟叶水分、叶绿素、多酚氧化酶、丙二醛等指标变化规律,比较烤后烟叶经济性状。结果表明:在施氮量为165 kg/hm2条件下,不同供氮形态对烤烟烘烤特性有不同影响。其中,30%氨态氮+70%硝态氮处理的烟叶在烘烤过程中各阶段失水速率接近其72 h内失水速率的平均值,烟叶失水均衡性好;其叶绿素降解速率与失水速率协调同步,有良好的变黄特性;同时,烟叶多酚氧化酶活性最低,丙二醛积累少且缓慢,膜脂过氧化水平最低,烟叶耐烤性好。此外,30%氨态氮+70%硝态氮处理的中上等烟比例和产值最高。可见,在粤北烟区,采用30%氨态氮+70%硝态氮的供氮形态可以获得优良的烟叶烘烤特性和烟叶质量。     

苜蓿接种根瘤菌及丸衣化效果研究初报

种子丸衣化可以提高种子的发芽率以及提高豆科植物的共生固氮作用,对改良退化草地,促进农业可持续发展的起着重要意义。实验选用中苜一号、美国（ML323）、准葛尔和20号4种紫花苜蓿作为实验材料,比较了苜蓿接种根瘤菌的效果。结果表明：在接种“先科”菌剂的比较试验中,美国（ML323）在其接瘤率、瘤重,及产量等方面均有较好的效果。     

不同硝态氮供应水平对平邑甜茶生长及氮素吸收利用和分配特性的影响

氮的吸收和合理分配对果树生长发育和产量及品质的形成具有重要作用,适宜供氮可实现植物的最适生长,充分挖掘植物的生长潜力。以平邑甜茶幼苗为试材,利用~(15)N同位素示踪技术,在砂培盆栽条件下研究不同硝态氮供应水平(0.00、4.00、8.00、12.00、16.00、20.00、24.00 mmol/LNO_3~-,分别用N_0、N_1、N_2、N_3、N_4、N_5、N_6表示)对植株生长及植株氮素吸收、利用和分配特性的影响。结果表明:随着供氮水平的提高,平邑甜茶幼苗的生物量、叶绿素(SPAD)、叶面积、根系长度、根表总面积、根尖数、根系活力、植株全氮均是先升高后降低,分别在N_3处理达到最高水平;Ndff值、~(15)N吸收量、氮肥利用率随着供氮水平提高均是一直降低。在低氮(N_1~N_2)条件下,~(15)N主要分布在叶片,N_1、N_2处理分别51.39%、45.90%,其次是根、茎。高氮(N4~N6)条件下,~(15)N开始向根部转移,N_4、N_5、N_6处理分别占43.80%、45.40%、51.46%。综合考虑在保证氮肥利用率较高的同时,Hoagland营养液硝态氮浓度在12.00 mmol/L水平时,显著促进了平邑甜茶幼苗的生长、根系的发生和氮素的吸收,各器官氮素分配比较均匀,平邑甜茶幼苗生长最适。     

Characterization of novel inbred lines of white clover (Trifolium repens L.). I. Dynamics of plant growth and nodule development in flowing solution culture

Growth, dry matter partitioning between shoots and roots, and extent of nodulation were characterised in four novel self-fertile highly inbred lines (referred to as A, B, C and D) of white clover (Trifolium repens L.) thought to differ fundamentally in their nitrogen relations and therefore regarded as candidate material for genetic mapping. Plants were inoculated with a mixture of three strains of Rhizobium and grown in flowing nutrient solutions without N for 18 d. Half the plants were then supplied with 20 μM NO₃ ^- during a 36 d treatment period, the remaining plants acting as ‘controls’ solely dependent upon N₂ fixation for acquisition of N. Total dry matter production and shoot:root ratios were similar in all lines supplied with NO₃ ^- and in control plants of lines B and C. Growth was severely reduced in control plants of line D and to a lesser extent in control line A. These effects were attributed to a failure to develop effective N₂ fixation capacity after apparently normal infection. Mean nodule size, nodule numbers and nodule d.w. per plant were extremely low in line D compared with the other lines, irrespective of whether NO₃ ^- was supplied. Lines A, B and C differed in the severity with which NO₃ ^- decreased mean d.w. per nodule and total nodule d.w. per plant relative to corresponding control plants, with line C being least sensitive. Nitrate also decreased the total number of nodules per plant relative to control plants after day 14 in all lines except C, and most severely in line B. This revised version was published online in August 2006 with corrections to the Cover Date.     

缺磷对已结瘤大豆生长和固氮功能的影响

磷对大豆生长起着非常重要的作用,然而区别磷在大豆生长和根瘤功能中的影响非常困难。为阐明根瘤形成后缺磷对大豆植株生长和固氮功能的影响,采用水培方法,营养液中供给正常磷酸盐浓度(30 µmol L^-1)条件下,待根瘤形成后,将营养液中的磷水平分别转换为0 µmol L^-1 (无磷)和4 µmol L^-1 (缺磷),研究缺磷对大豆生长和结瘤固氮的影响。结果表明,大豆根瘤具有固氮功能后,进行营养液中不同浓度的磷酸盐处理,从处理第9天开始,缺磷对大豆生物量的影响才表现出来,而缺磷对根瘤形成和生长的影响从处理开始就比较明显。处理第9天,无磷、缺磷和正常磷处理根瘤数比处理第3天分别增加11.8%、15.4%和20.0%,单位植株豆血红蛋白含量和单位根瘤豆血红蛋白含量都随磷浓度增加而增加。该结果表明,在保证大豆形成一定数目的根瘤后,缺磷会明显影响根瘤生长和固氮能力。     

Nitrogen Fixation in Hybrids of White Clover (Trifolium repens L.) and Caucasian Clover (Trifolium ambiguum M. Bieb)

Hybrids between white clover (Trifolium repens L.) and Caucasian clover (or Kura clover, Trifolium ambiguum M. Bieb) are a potential route for the improvement of drought tolerance and persistence in white clover. However, to be agronomically viable they must show no significant reduction in their potential for nitrogen fixation relative to white clover. A comparative study of growth rate and nitrogen fixation was carried out in flowing solution culture without a supply of mineral nitrogen to the plants. The two parental species and two generations of backcross hybrids, with white clover as the recurrent parent, were assessed. The growth rate and N content of T. ambiguum were significantly lower than those of the other lines. However, dry matter production, nodule biomass per plant and rates of fixation were similar in second‐generation backcross plants and white clover. The results suggest that the agronomic potential of this novel germplasm is not compromised by limitations with respect to nitrogen fixation.     

三江平原草甸小叶章群落对不同形态氮的吸收偏好

为探讨三江平原植被对不同形态氮的吸收偏好,以三江平原典型植物群落——草甸小叶章群落为研究对象,利用¹⁵N示踪技术,施加标记的NH⁺₄-N和NO^-₃-N。结果表明：草甸小叶章群落5种植物均表现出喜硝性,且优势种小叶章吸收的铵态氮和硝态氮最多。优势种小叶章不同器官短期内偏好硝态氮的优先顺序为叶>茎>根。土壤中大部分外源氮被植物吸收,土壤残留的铵态氮多于硝态氮。     

Interactive Effects of Salinity and Biological Nitrogen Fixation on Chickpea (Cicer arietinum L.) Growth

The effect of salinity on the nodulation, N-fixation and plant growth of selected chickpea- Rhizobium symbionts was studied- Eighteen chickpea rhizobial strains were evaluated for their growth in a broth culture at salinity levels of 0 to 20 dS m⁻¹ of NaCl + Na₂SO₄. Variability in response was high. Salinity generally reduced the lag phase and/or slowed the log phase of multiplication of Rhizobium. Nine chickpea genotypes were also evaluated for salt tolerance during germination and early seedling growth in Petri dishes at five salinity levels (0–32 dS m⁻¹). Chickpea genotypes ILC-205 and ILC-1919 were the most salt-tolerant genotypes. The selected rhizobial strains and chickpea cultivars were combined in a pot experiment aimed at investigating the interactive effect of salinity (3, 6 and 9 dS m⁻¹) and N source (symbiosis vs. inorganic N) on plant growth. Symbiotic plants were more sensitive to salinity than plants fed mineral N. Significant reductions in nodule dry weight (59.8 %) and N fixation (63.5 %) were evident even at the lowest salinity level of 3 dS m-¹. Although nodules were observed in inoculated plants grown at 6 dS m-¹, N-fixation was completely inhibited. The findings indicate that symbiosis is more salt-sensitive than both Rhizobium and the host plant, probably due to a breakdown in one of the processes involved in symbiotic-N fixation. Improvement of salinity tolerance in field grown chickpea may be achieved by application of sufficient amounts of mineral nitrogen.     

盐胁迫对苜蓿种子发芽性状的影响及其耐盐性评价

为了阐明盐胁迫条件下紫花苜蓿种子的发芽性状表现和种子萌发期耐盐性能强弱,同时为改良盐碱化土壤提供更加耐盐碱的植物品种,提高盐碱化土地资源利用率,本试验以5个苜蓿(Medicago sativa)品种为材料,采用不同浓度NaCl处理(0、0.4%、0.8%、1.2%),研究了不同盐浓度对苜蓿种子发芽率、发芽势、苗高、根长的影响。结果表明,盐胁迫对苜蓿种子发芽性状均有较大影响,随着NaCl浓度的提高,苜蓿种子发芽率和发芽势降低,苗长和根长明显缩短,而且,高质量浓度盐胁迫对苜蓿种子发芽性状的影响尤为显著。不同苜蓿品种在不同盐浓度间的差异显著;品种间各指标的表现不尽相同,发芽势与发芽率指标在品种间的差异显著,苗高和幼根长指标在品种间的差异不显著。综合单项指标耐盐系数和品种综合耐盐指数来看,5个苜蓿品种的耐盐性能强弱顺序为‘Magnum salt’>‘中苜1号’>‘维多利亚’>‘巨人201T’>‘皇冠’。     

Selection of High Nitrogen-Fixing Rhizobia Nodulating Chickpea (Cicer arietinum) for Semi-Arid Tunisia

Inoculation of grain legumes with rhizobia may improve biological N₂ fixation and crop yield. However, drought, high temperature and soil salinity constrain legume root-nodule formation and function. Here, two rhizobial strains nodulating Tunisian chickpea, Mesorhizobium ciceri strain CMG 6 and Mesorhizobium mediterraneum strain CTM 226 originating from semi-arid regions, were selected for their symbiotic performance and their salt stress tolerance (3 % NaCl). Both strains were then examined as inoculants in different soils and field conditions. Field experiments were conducted in four sites using four chickpea cultivars. Rhizobia occupying nodules in non-inoculated plots were isolated and characterized using 16S rDNA typing; to examine nodule occupancy by the inoculant strains we used polymerase chain reaction (PCR)-restriction fragment length polymorphism of 16S rDNA gene and repetitive extragenic palindromic PCR. The inoculant strains gave a significant increase in nodule number, shoot dry weight and grain yield in all the experimented fields for the four cultivars used, even in the non-irrigated soils. The improvement in plant production was equal to or better than nitrogen fertilization. Moreover, the monitoring of the nodule occupancy showed that inoculant strains competed well in the native populations of rhizobia. These results suggest that nodulation and yield of chickpea can be improved by inoculation with competitive and salt-tolerant rhizobia and is economically promising to increase chickpea production in water-limited regions.     

Karyotypic and electrophoretic studies on taro and its origin

Summary Karyotypes and electrophoretic pattern of 15 strains of taro were studied. Strains collected from the northeast India hill state, Meghalaya were diploids and triploids whereas those from the plains of south India were diploids and of north India was a triploid. The diploids had 2n=28 and triploids showed 2n=42 chromosomes. The wild taro had the most asymmetrical karyotype. The protein content varied from 4.2 to 11.4 mg/g dry wt. The maximum protein content was found in a triploid strain 8 (11.4 mg/g dry wt) and minimum in the wild taro (4.2 mg/g dry wt). The number of protein bands was 7 in the wild taro (diploid) and 12 in one of the cultivated triploid strain. Meghalaya strains showed great variation with respect to leaf size and tuber shape and size. All the strains have diverged at morphological, karyotypic and genotypic levels. It is suggested that taro might have originated in the north-eastern India.     

Mineral Nitrogen Availability and Isoflavonoid Accumulation in the Root Systems of Soybean (Glycine max (L.) Merr.)

Isoflavonoids, as plant-to-bacteria signal molecules, play an important role in the establishment of the soybean ( Glycine max (L.) Merr.)– Bradyrhizobium japonicum nitrogen (N) fixing symbiosis. They are essential to the development of effective root nodules and responsible for inducing the nod genes of B. japonicum . Because N affects a broad range of infection events, especially the symbiotic events occurring within 18 h of inoculation, it is reasonable to hypothesize that mineral N disrupts the interorganismal signal exchange between soybean host plants and B. japonicum . High performance liquid chromatographic (HPLC) analysis of root extracts of soybean, inoculated with B. japonicum or not, grown with various levels of mineral N in the rooting medium were performed to test this hypothesis. The results of these studies indicated that: (1) at early plant growth stages (before the onset of N fixation), a strong negative relationship between N application and soybean root isoflavonoid (genistein and daidzein) concentrations existed; (2) although isoflavonoid (genistein and daidzein) concentrations in both inoculated and non-inoculated soybean root systems were generally decreased by N application, at very low N levels (10 mg N l⁻¹) genistein in the non-inoculated plant roots was not decreased relative to the 0 N plants; (3) averaged over all mineral N treatment levels and sampling times, inoculation of soybean with B. japonicum increased root daidzein concentrations (P > 0.05), but did not affect genistein. Overall, N application reduced the isoflavonoid concentration of soybean root systems, which probably plays a part in the regulation of soybean nodule formation by available N.