茎瘤固氮根瘤菌Azorhizobium caulinodansORS571基因组分泌蛋白的生物信息学分析

为了获取茎瘤固氮根瘤菌（AzorhizobiumcaulinodansORS571）的分泌蛋白,以便更深入地了解该菌的共生固氮作用,本研究采用SignalP、TMHMM、PSORTb、TargetP、LipoP、TatP和SecretomeP软件对该菌全部4717个蛋白序列进行分析预测。结果共识别了653个分泌蛋白,其中具有分泌型信号肽的蛋白54个,具有RR—motif型信号肽的蛋白1个,具有脂蛋白信号肽的蛋白2个和非经典分泌蛋白596个。该菌含信号肽分泌蛋白仅占全部蛋白的1．2％,低于其它固氮菌。在分泌蛋白中识别了核酸内切酶和核糖核酸酶等6个核酸酶。它们可能参与宿主植物遗传物质的降解,干扰宿主遗传代谢,进一步在宿主植物侵染过程中起到重要作用。此外还识别了超氧化物歧化酶、过氧化氢酶和谷胱甘肽S-转移酶等4个抗氧化酶。它们可能参与活性氧的清除以保护固氮酶,是该菌固氮过程的重要参与者。     

豇豆叶绿体基因组密码子使用偏好性分析

为了探究豇豆（Vigna unguiculata）叶绿体基因组密码子的使用模式,本研究从NCBI下载完整的豇豆叶绿体基因组序列,并对其进行结构分析。利用CodonW和CUSP对筛选获得的50条可编码蛋白序列（CDS）进行分析,获得GC1、GC2、GC3、RSCU、CAI、CBI、Fop、ENc、RFSC等重要参数,并进行中性绘图、PR2-plot绘图、ENc-plot绘图以及对应分析、最优密码子分析和其他物种对比分析。结果发现,豇豆叶绿体基因密码子更偏向以A或U(T)结尾,G和C在密码子各位置中的占比较低,平均值为36.31%;有效密码子数（ENc）的平均值为44.903,密码子偏好性较弱;GC1与GC2、GC3间均有相关性,表明碱基突变对密码子选择也有影响。从中性绘图、PR2-plot绘图、ENc-plot绘图结果可以看出,豇豆叶绿体密码子使用偏好性同时受到了碱基突变与自然选择的影响。本研究最终筛选出20个最优密码子,并将豇豆叶绿体基因组密码子使用频率与其他物种进行比较,发现豇豆与番茄在密码子使用频率上存在较高的相似度。本研究结果为提高豇豆叶绿体外源基因的表达效率提供了参考依据。     

蒺藜苜蓿WRKY转录因子密码子使用偏好性分析

密码子使用偏好性普遍存在于各种生物中,但突变和自然选择压力对每种生物密码子使用偏好性不同.WRKY基因家族是一类只存在于植物的转录因子,主要参与植物体内转录调控和信号转导过程.本研究以蒺藜苜蓿(Medicago truncatula)WRKY转录因子(MtWRKY)为研究对象,揭示Mt WRKY基因密码子使用偏好性形成的主要因素,并筛选最优密码子.研究结果表明,代表MtWRKY基因的点均分布在有效密码子数(effective number of codons,ENC)标准曲线以下,表明密码子受自然选择压力或突变选择压力或其他因素影响;密码子第1、2位平均GC含量(GC12)与密码子第3位GC含量(GC3)相关性分析发现,GC12与GC3呈显著正相关(r=0.34,P＜0.01),表明突变压力导致密码子3个位点具有相似的GC含量;GC3s值分布在0.2～0.5之间,表明密码子使用偏好性主要受突变压力影响.奇偶偏好分析表明,MtWRKY基因第3位密码子CT含量＞AG含量.G和C(或者A和T)不成比例分布在密码子第3位上,表明密码子使用偏好性受到自然选择压力影响,但很可能突变压力仍起主要作用.最优密码子使用频率(frequency of optimal codons,Fop)与GC含量以及序列长度相关性分析发现,Fop与外显予GC含量呈显著正相关(r=0.57,P＜0.01),而与内含子GC含量呈较弱正相关(r=0.09,P＞0.05);Fop与外显子序列的长度呈正相关(r=0.28,P＜0.05),而与内含子长度呈负相关(r=-0.01,P＞0.05).表明MtWRKY基因外显子和内含子序列的形成受不同选择压力影响;外显子密码子使用偏好性受突变压力影响,而内含子可能是由于自然选择压力作用于突变选择形成的.确定了4个以G或C结尾的最优密码子.研究结果为WRKY转基因研究过程中密码子优化提供了理论支持.     

欧洲葡萄碱性螺旋-环-螺旋(bHLH)基因家族密码子使用偏性分析

碱性螺旋-环-螺旋(basic helix-loop-helix,bHLH)转录因子作为植物转录因子最大家族之一,在植物生长发育、胁迫应答和植物次生代谢中具有重要的调控作用。本研究利用CodonW及SPSS等软件,计算和统计分析了欧洲葡萄(Vitis vinifera)bHLH基因家族106条序列密码子的使用模式,并对影响该基因家族密码子组成及编码特点的主要因素进行了探讨。结果表明,bHLH基因家族密码子GC1与GC2之间的相关系数达到极显著水平(P0.01),GC3与GC1、GC2的相关系数经双尾检验均未达到显著水平(P0.05);ENC与GC1、GC2和GC3的相关系数分别为0.1439、0.0463和0.1766,均未达到显著水平(P0.05),ENC和GCtotal的相关系数为0.1989,达到显著水平(P0.05);确定了19个最优密码子,多以G或C结尾;推测该基因家族密码子的偏性可能是由选择和突变共同影响的。研究结果为深入开展葡萄属(Vitis)植物bHLH基因家族分子进化、表达调控机制和提高该基因家族成员预测的准确性以及进一步应用该基因家族成员进行葡萄抗逆改良等提供了重要的理论依据。     

Ammonium fixation from urea as influenced by nitrapyrin

Abstract

During the period 1977–1979, NaNO₃, urea, and urea plus 2% (wt/wt) nitrapyrin (2‐chloro‐6‐(trimethyl)pyridine) were compared on a Matapeake silt loam (fine silty mixed mesic Typic Hapludult) . Nitrogen sources were injected as solutions into the water system at 224 kg N ha^‐1yr^‐1used for subsurface trickle irrigation of corn (Zea maysL.). Nitrogen was withheld in 1980 in order to assess residual N effects. Grain yields in 1980 for the NaNO₃, urea, and urea plus Nitrapyrin treatments were 5.10, 4.56 and 6.52 Mg ha^‐1, respectively. Corresponding ear leaf N concentrations were 17.7, 16.7 and 19.2 g kg^‐1. Significantly higher grain yield and leaf N concentrations associated with the use of nitrapyrin as a nitrification inhibitor indicated greater soil N reserves for this treatment. Non‐exchangeable (fixed) NH₄ ⁺, in soil cores taken in November 1981 averaged 54, 59 and 74 ug N g^‐1for the respective N regimes. The concentration of fixed NH₄ ⁺increased with sampling depth, averaging 54, 61 and 72 ug N g^‐1for the 0–5, 30–35, and 60–65 cm profile depths, respectively. This trend is ascribed to increasing quantities of micaceous and vermiculitic clay (<2 um) with increasing profile depth.     

Nodulation and Symbiotic Nitrogen Fixation of Cowpea Genotypes as Affected by Fertilizer Nitrogen

Abstract

Intercropping with legumes and non‐legumes is commonly practiced in many parts of the world to maximize productivity per unit area of land. In India, cowpea [Vigna unguiculate (L.) Walp] is a popular pulse legume component of intercropping farming systems. Often, however, potential production is compromised, particularly in high fertilizer input systems, because legume component competes with the non‐legume component of the system for nitrogen (N) in the soil. An experiment was conducted in order to identify lines of cowpea that could obtain the majority of their nitrogen requirements from symbiotic fixation of atmospheric nitrogen rather than from uptake of soil nitrogen. Twenty‐nine genotypes of cowpea were screened for tolerance to (applied) nitrogen in soil in field condition. The parameters used to appraise tolerance were extent of root nodulation, the amount of nitrogen fixed, nitrate reductase activity (NR) in roots and nodules, and nitrite content of roots and nodules. There were two nitrogen treatments applied as urea, 40 kg N per ha (N40), and 120 kg N per ha (N120). There were three genotypes whose nitrogen‐fixing effectiveness was apparently unimpaired by applications of nitrogen to the soil. Genotype EC‐170442‐3 nodulated and fixed atmospheric nitrogen satisfactorily at higher levels of applied nitrogen. At N40, genotypes EC‐244390 and EC‐240900 formed a great abundance of large nodules effective in nitrogen fixation; even at N120, EC‐240900 had better symbioses than the majority of the 29 cowpea lines originally screened. These three genotypes are deemed worthy of further examination for their suitability for intercropping systems. How this might be achieved is discussed.     

Legumes as nitrogen suppliers of grassland on fen soil: Leguminosen als stickstofflieferanten für niedermoorgrünland

A 6-year trial was laid out with varied PK fertilization on fen soil and analysed for the capability of undersown clover species to fix atmospheric nitrogen. No mineral nitrogen was applied. On an average of the six test years, the share of white clover in the total crop yield of the sward was 17% after sowing once and that of red clover or Swedish clover was 19% after sowing twice. The annual mean shares of these two clover variants in the total yield varied only slightly over the test period. White clover showed typical seasonal dynamics in the test years, with yield portions increasing from spring to late summer. Undersown red and Swedish clovers reached the highest yield percentages in their 2nd and 3rd growths. The variants grown without PK fertilizer and clover yielded 45 dt DM ha^???1. With PK fertilizer dry matter yields increased to 57 and 68 dt ha^???1, respectively. With supplemental white clover or red/Swedish clover sown into the sward, substantial amounts of symbiotic nitrogen were accumulated in the harvestable plant matter, giving a total dry matter yield of 76 dt ha^???1. Clover yield shares up to 30% resulted in increasing DM yields which, however, declined again with even larger shares of clover. As to the ingredients of grasses, herbs and clover, the differences were mostly significant. On an average of the six test years, nitrogen fixation was 58.4?kg ha^???1 for white clover and 46.6?kg ha^???1 for red/Swedish clover.     

不同基因型大豆氮效率的研究

通过对不同基因型大豆新大豆1号和黑农40的试验研究,结果表明,施氮对新大豆1号的增产作用不明显,但对黑农40有较好的增产作用;花期黑农40和新大豆1号上、中、下层叶片氮含量都随着施氮量的增加而增加,黑农40叶片氮含量与施氮量的相关系数都达到极显著相关,但新大豆1号没有达到显著水平;施氮对新大豆1号的生物固氮抑制程度高于黑农40,使新大豆1号生物固氮降低36. 3% ~85. 2%。     

The glycine‐glomus‐rhizobium symbiosis III. Endophyte effects on leaf carbon,nitrogen, and phosphorus nutrition

Soybean plants [Glycine max (L.) Merr.] were grown in pots and inoculated with Rhizobium japonicum and/or Glomus mosseae (Nicol. & Gerd.) Gerd. & Trappe, either at planting or 20 days later. Nitrogen was supplied in the nutrient solution to plants without nitrogen‐fixing bacteria, and P was added to those without the mycorrhizal fungus. At harvest, 50 days after planting, all plants had leaves of similar dry mass. Each root symbiont grew best in the absence of the other. Growth of Glomus reflected the duration of its growing time and the presence and duration of competition from Rhizobium. Nodule weight in the tripartite associations, on the other hand, was inhibited only by the earlier introduction of Glomus.

Dipartite associations and the plants inoculated with both root symbionts at planting had the highest concentration of leaf N, and the lowest was in those inoculated with both organisms at d 20. Leaf P was highest in plants inoculated only with Rhizobium, and lowest in those tripartite associations involving any inoculation at day 20. The low values were presumably a result of the short duration of endophyte‐mediated P uptake before the plants were harvested.

Although there was almost no difference in leaf sugar concentrations, starch concentrations reflected the duration of Glomus growth, and were greatest in those plants that had supported it for the least time. Uninoculated plants contained the least starch, but produced a greater fresh mass of leaf tissue than any of the tripartite symbionts.     

烤烟基肥施用时间与氮肥利用的相关性研究

烤烟是一种氮素敏感型的品质作物, 对氮素的需求非常严格。氮肥的合理施用不仅有利于提高烤烟对氮素养分的吸收利用, 也有利于提高烤烟烟叶品质。本研究采用大田试验的方法, 在湖北省烤烟主产区研究了基肥在3个不同施用时间(烤烟烟苗移栽前0 d、15 d和30 d)对烤烟氮肥利用参数的影响及其相关性。结果表明, 烤烟地上部烟叶和茎秆中干物质累积量和氮素含量均随着基肥施用时间的提前而表现出增加趋势。与移栽当天(0 d)施用基肥相比, 提前15 d、30 d施用基肥, 烤烟烟叶中干物质累积量和氮素含量分别平均增加16.4%、22.6%和1.5%、8.7%; 茎秆中则分别平均增加2.5%、12.7%和13.4%、33.9%。提前施用基肥有提高烤烟氮肥表观利用率的趋势, 但是各处理间差异不显著(P>0.05), 而在烤烟烟苗移栽前15 d、30 d施用基肥显著降低烤烟地上部氮肥农学利用率(35.2%、37.4%)和生理利用率(35.4%、41.6%)(P<0.05)。另外, 基肥施用时间与氮肥农学利用率、生理利用率和偏生产力均表现出显著或者极显著负相关关系, 而氮肥农学利用率与生理利用率和氮肥偏生产力以及氮肥生理利用率与偏生产力之间均表现出显著或极显著正相关关系。因此, 在烤烟烟苗移栽时适当提前基肥的施用时间(15~30 d)有利于提高烤烟对氮素的吸收利用, 促进烟叶中干物质的累积。     

Correlation of soil test nitrogen with potato yields

Abstract

Yield response of Idaho Russet Burbank potatoes to nitrogen fertilizer was related to soil test for inorganic N (NO₃ ^‐ and NH₄ ⁺) in a total of 27 field experiments over a 3‐year period using polynomial correlation and regression analysis. Nitrate plus ammonium nitrogen content in the surface foot of soil was found to be useful in predicting yield response of potatoes to applied nitrogen.

Correlations between yield and extractable N were considerably better when the data from each cropping system were analyzed separately than when all locations were analyzed as one group. Additional improvement was obtained by including extractable ammonium nitrogen and nitrogen in the second foot of soil respectively. The best correlation with yield was found using (NO₃ ^‐ + NH₄ ⁺)‐N in the surface foot following grain and the top two feet following non‐grain crops with R² values of 0.875 and 0.821 respectively.     

Nodulation of African yam bean (Sphenostylis stenocarpa) by Bradyrhizobium sp. isolated from Erythrina brucei

African yam bean (Sphenostylis stenocarpa), which is widely cultivated in Africa because of its growth capability on marginal soils, was nodulated by an endosymbiont (characterized and designed Bradyrhizobium sp. AUEB20) isolated from the Ethiopian tree Erythrina brucei with the formation of a small number of large, indeterminate N₂-fixing nodules. In contrast, 24 other isolates from Ethiopian woody legumes were ineffective. Strain AUEB20 promiscuously nodulated a number of tropical legumes, but none out of five European crop plants tested. Received: 17 September 1996