缺硼对大豆植株生长和根瘤固氮活性的影响

以普通结瘤大豆Ｂｒａｇｇ ｃｖ．（Ｇｌｙｃｉｎｅ ｍａｘ（Ｌ．）Ｍｅｒｒ．）及其超结瘤突变体ｍｔｓ３８２为试验材料,用营养液培养方法研究了缺硼对大豆植株生长和根瘤固氮作用的影响,结果表明,在缺硼条件下,植株根系生长受到抑制,变粗变短,颜色加深;地上部生长点坏死,使植株长得矮小,干重降低;根系结瘤能力和根瘤固氮活性明显下降,说明硼不仅对维持植株的正常生长,同时对根瘤的形成及其固氮活性都起着重要作用。     

施氮对大豆根瘤生长和结瘤固氮的影响

采用框栽试验研究了施氮对大豆根瘤生长和结瘤固氮的影响,结果表明,施氮对大豆根瘤形成、生长和固氮能力有显著影响,随着N用量的增加,根瘤干质量、根瘤数量呈现先逐渐增加而后降低的趋势,而固氮酶活性和豆血红蛋白含量则表现为持续下降的趋势,适量施氮对根瘤生长有显著的促进作用,当氮素供应不足时则会抑制根瘤的生长,但当氮素供应过量时也会抑制根瘤的形成.从根瘤干质量和根瘤数量来看,各处理间表现为N100>N200>N50>N25>NO,以N100处理下根瘤干质量最大,根瘤数量最多,显著高于不施氮(N0)和其他施氮处理,从不同生育时期来看,根瘤干质量表现为从苗期到花期再到鼓粒期,大豆根瘤的数量呈现出先明显增加后逐渐减少的趋势,高峰出现在花期,而根瘤数量表现为花期>苗期.施氮显著抑制了固氮酶活性和豆血红蛋白含量,随施氮水平的增加,固氮酶活性和豆血红蛋白含量显著降低,表现为NO>N25>N50>N100>N200,表明施氮使大豆根瘤的固氮效率显著降低,因此,从大豆固氮效率来讲,施氮对大豆根瘤固氮具有抑制作用.方差分析结果表明,施氮和不施氮处理间均达到了5%的差异显著水平.因此,从大豆固氮和氮肥施用平衡的角度来看,应适量施用氮肥,既可以充分利用大豆的固氮功能,节约氮肥,又可以获得较高的产量.     

氟磺胺草醚对大豆根瘤固氮和蔗糖代谢的影响

采用田间试验方法研究了氟磺胺草醚茎叶处理对大豆根瘤固氮酶和蔗糖代谢的影响。结果表明:不同剂量的氟磺胺草醚茎叶处理对大豆根瘤生长无影响。氟磺胺草醚525g a.i./hm²用量下,根瘤固氮酶活性在施药后的38d内始终受到显著抑制,叶片和根中蔗糖合成酶活性受到长时间的抑制,同时叶片、茎、根系和根瘤中蔗糖含量均显著降低;263g a.i./hm²用量下,根瘤固氮酶活性在施药后的24d内显著降低,对叶片和根中蔗糖合成酶活性抑制作用最高值分别出现施药后的第10天和第18天,酶活性与对照相比分别降低15.22%和11.76%。氟磺胺草醚茎叶处理后,叶片、茎、根系和根瘤中蔗糖含量显著降低,蔗糖向根瘤中的供应不足是造成大豆根瘤固氮酶活性降低的一个重要原因。     

NO与Ca2+对蚕豆保卫细胞气孔运动的互作调控

以蚕豆(Vicia faba L.)为材料研究NO和Ca²⁺对蚕豆气孔运动及质膜K⁺通道的影响。结果表明,10 mmol L^-1 Ca²⁺和100 µmol L^-1 NO供体SNP均有效抑制气孔开放,NO清除剂c-PTIO不能缓解Ca²⁺抑制气孔开放,相反胞外加入0.1 mmol L^-1 Ca²⁺可以明显加强NO对气孔开放的抑制程度,该现象可被La³⁺(Ca²⁺通道抑制剂)缓解。以膜片钳技术记录全细胞K⁺电流发现,胞外10 µmol L^-1或100 µmol L^-1 SNP均可选择性抑制蚕豆保卫细胞质膜内向K⁺通道,追加0.1 mmol L^-1 Ca²⁺可显著激活质膜外向K⁺通道,且可被La³⁺所缓解,然而0.1 mmol L^-1 Ca²⁺单独作用并不影响质膜外向K⁺通道活性。10 mmol L^-1 Ca²⁺单独处理可激活质膜外向K⁺通道,但不能被c-PTIO缓解。分别用Ca²⁺和NO专一的荧光探针Fluo-3-AM和DAF-2DA标记蚕豆保卫细胞原生质体,检测胞内Ca²⁺和NO的水平变化发现,100 µmol L^-1 SNP明显诱导胞内Ca²⁺积累,但10 mmol L^-1 Ca²⁺并不能诱导NO在细胞内积累。记录保卫细胞质膜Ca²⁺通道电流发现,NO可明显激活质膜Ca²⁺通道。表明NO有效抑制气孔开放,可能主要通过激活质膜Ca²⁺通道,提高胞内Ca²⁺,激活质膜外向K⁺通道促进K⁺外流,同时,可选择性抑制内向K⁺通道阻止K⁺内流,两种途径共同作用抑制气孔开放。然而,胞外10 mmol L^-1 Ca²⁺对气孔和质膜K⁺通道活性的调节并不依赖于NO。     

种间距对不同结瘤特性套作大豆根瘤生长及固氮潜力的影响

为充分挖掘套作大豆的固氮潜力,本试验以玉米/大豆带状套作系统为对象,研究了不同玉豆间距(30 cm、45 cm、60 cm和75 cm)下不同结瘤特性大豆品种(贡选1号、桂夏3号和南豆25号)的根瘤生长及固氮潜力差异。结果表明,与单作相比,带状套作可延缓大豆根瘤数目和鲜重峰值出现的时间:玉豆间距60 cm处理下,各品种大豆根瘤数目和鲜重显著高于其他间距处理,并在达到峰值期后高于单作大豆;品种间表现为:南豆25号>桂夏3号>贡选1号。带状套作相对于单作会降低始粒期(R5)前大豆根瘤的单株固氮潜力,但玉豆间距60 cm处理下, R5期贡选1号、桂夏3号和南豆25号的单株固氮潜力2年平均较单作提高8.53%、16.40%和13.70%。不同大豆品种根瘤侵染细胞内含物积累过程差异较大,相较单作,玉豆间距60cm处理下,R5期类菌体数量增多,其中聚-β-羟基丁酸盐颗粒(PHB)增加,以南豆25号表现最优。因此,带状套作下适宜的种间距(玉豆间距60cm)可增加R5期大豆根瘤数目和鲜重,提高大豆根瘤类菌体和PHB的数量,增强大豆根瘤的固氮潜力,以强结瘤品种南豆25号效果最好。     

马铃薯适应低磷胁迫的生理生化效应

以马铃薯两个栽培种大西洋和新大坪为材料,采用基本Hoagland营养液和60%磷浓度(0.05996 μmol/L)的Hoagland营养液处理,研究了低磷胁迫效应对马铃薯植株生长、保护酶活性和叶绿素含量影响.结果表明,60%磷胁迫对马铃薯地上部分生长具有抑制作用,能促进新大坪侧根生长和根冠比增加,而大西洋在根系生长和...     

铝胁迫下大豆根尖细胞铝的微区分布与耐铝性分析

以浙春3号为实验材料, 利用透射电镜(TEM: Transmission Electron Microscope)-X-射线能谱(EDS: Energy Dispersive X-ray), 调查铝胁迫下大豆根尖铝的微区分布及耐铝性。结果表明,Al³⁺胁迫导致根尖细胞细胞壁不规则加厚, 线粒体数量增多, 核膜膨胀, 液泡中存在较多的电子致密沉淀物。90 mg L^-1 Al³⁺处理的根尖细胞内含物完全降解消失, 仅剩细胞壁。10 mg L^-1 Al³⁺处理的线粒体、细胞壁和液泡电子致密沉淀物中均检测到Al;随着Al³⁺处理浓度的增大, 各细胞器中Al的质量和原子数百分比逐渐增大。线粒体在60 和90 mg L^-1Al³⁺处理下, 液泡电子致密沉淀物在90 mg L^-1Al³⁺处理下,均未被检测出Al。在60 mg L^-1Al³⁺处理下唯一一次在细胞核中检测到Al。Al³⁺抑制了根系生长, 根系细胞中细胞壁的Al³⁺含量受影响最明显。P/Al在细胞壁和线粒体中的相对原子数随Al³⁺浓度的增大而下降。研究结果表明X–射线能谱对铝在亚显微结构上的定位是一种快速、有效的方法。铝最先积累在细胞壁上, 随Al³⁺处理浓度增大逐渐积累于部分细胞器和细胞核中, 且含量在细胞中的分布亦由外向里呈递减趁势。     

Na2CO3对黄秋葵种子萌发及生理特性的影响

为研究碱环境下黄秋葵种子萌发及生理特性,采用不同浓度(0、10、20、30、40 mmol·L^-1)Na2CO3溶液处理黄秋葵种子,测定其种子的萌发特性、胚根生长速度及第3天时胚根的生理特性。结果表明,随着碱浓度的增加,种子的发芽率、发芽势、发芽指数呈下降趋势,相对碱害率升高;发芽期间胚根长度与碱浓度呈负相关关系,且生长速度降低;碱胁迫下种子的抗氧化物酶(POD、CAT、SOD)活性较对照组无显著变化,MDA随碱浓度增加呈升高趋势,脯氨酸积累量变化不明显。发芽7 d后对照和10 mmol·L^-1碱液处理可萌发为幼苗,对其复水5 d后,发现10 mmol·L^-1处理侧根生长量远少于对照组。说明碱胁迫能抑制黄秋葵种子萌发及其幼苗生长,但黄秋葵种子对10 mmol·L^-1碱胁迫具有耐受性。     

铅胁迫对薏苡种子萌发和幼苗生理特性的影响

研究了铅胁迫对薏苡种子萌发和幼苗生长的影响。采用不同浓度的硝酸铅处理薏苡种子和幼苗,记录薏苡种子发芽率、发芽势、发芽指数和活力指数,并在幼苗生长期检测叶片相关生理指标。结果表明,铅胁迫抑制幼苗生长和活力指数,中高浓度(30~60 mg·L^-1)硝酸铅能够明显抑制种子萌发。铅胁迫下抑制幼苗叶片中叶绿素和类胡萝卜素合成,其中类胡萝卜素对铅胁迫更加敏感。随着硝酸铅浓度的升高,幼苗叶片中MDA、相对电导率和脯氨酸含量呈明显上升趋势;中低浓度(15 mg·L^-1和30 mg·L^-1)硝酸铅对SOD酶活性具有微弱促进作用,40 mg·L^-1和60 mg·L^-1硝酸铅对SOD酶活性具有明显抑制作用。研究表明,铅胁迫抑制薏苡种子萌发和幼苗生长,薏苡幼苗通过多种生理途径抵御铅胁迫。     

田间施加AM菌剂对大豆生长效应的影响

为进一步揭示在大田条件下施加丛枝菌根(AM)菌剂对作物生长的影响,以大豆为研究对象,田间播种时分别施加摩西球囊霉(Glomus mosseae,GM)和根内球囊霉(Glomus intraradices,GI)2种AM菌剂,以施加化肥处理(F)和既不施加菌剂也不施加化肥(CK)做为对照,研究AM菌剂对大豆生物量、根瘤数和土壤酶活的影响。结果表明：2种AM真菌在田间条件下都能侵染大豆根系形成菌根,GM处理的大豆菌根侵染率最高达到86.6%,GI处理的菌根侵染率最高达到78.3%。施加2种AM菌剂可显著提高大豆植株的生物量,GM、GI处理的大豆产量与CK相比分别提高了7.38%和3.88%。施加AM菌剂的大豆根瘤数显著高于CK处理,说明AM真菌能促进大豆根瘤的形成,从而提高大豆的固氮能力。施加AM菌剂可显著提高大豆根际土壤脲酶和过氧化酶的活性,且在大豆分枝期酶活性达到最大。另外,施加化肥能抑制脲酶的活性,使土壤中脲酶活性降低,而对过氧化氢酶活性的影响不显著。该研究结果为AM菌剂在田间推广应用奠定了理论基础。     

Variability of N2-fixation in common bean (Phaseolus vulgaris L.) under P deficiency is related to P use efficiency

Common bean often faces P deficiency in soils where it is generally grown. Such a deficiency is a major limitation to grain yield improvement, especially when common bean relies on N2 fixation. Screening for symbiotic N2 fixation (SNF) under P deficiency (72 μmol P plant-1 week-1) was performed in a glasshouse with 220 lines originating from the Andean and Meso-American gene pools. Large variability in shoot dry weight and SNF under P deficiency was found. SNF tolerance to P deficiency was mostly found among late flowering type IV lines, with the exception of three early flowering type III lines. The SNF tolerance to P deficiency was correlated with: (i) low nodule P concentration in all the growth types, and (ii) intense and early nodulation for type IV lines, or (iii) large root dry weight and high nodule nitrogenase for type III lines. Large genotypic variability for nitrogen use efficiency, i.e. biomass per N concentration was also found. However, the ratio plant N per plant P concentration was the most discriminating. We concluded that this later ratio, which is almost similar to plant N-fixed per plant P concentration, in glasshouse hydroponics culture might be: (i) an estimate of the P use efficiency for SNF, (ii) a consequence of low P concentration in plant organs, especially nodules, and (iii) a determinant of genotypic variability of SNF tolerance to P deficiency. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effect of N fertilizer top-dressing on N accumulation and N2 fixation of supernodulating soybean mutant

Increased application of nitrogen (N) fertilizer top-dressing during growth is an effective option for enhancing N supply to soybean plants. SS2-2 was characterized by the superior ability of symbiotic N₂ fixation at the level of 30 kg N ha⁻¹. But, the response of nitrogen fixation ability of supernodulating soybean mutant, SS2-2, to N fertilizer application rate remains unclear. The objective of this experiment was to compare the response of N fertilizer top-dressing on N accumulation and N₂ fixation between supernodulating mutant, SS2-2, and wild-type, Sinpaldalkong 2. The effect of N fertilizer top-dressing (0.6 g N pot⁻¹ top-dressing) on the nitrogen accumulation and redistribution were compared between SS2-2 and Sinpaldalkong 2. N fertilizer top-dressing at R1 stage increase in plant dry weight, relative growth rate (RGR), net assimilation rate (NAR), nitrogen harvest index (NHI), and N redistribution (NR). SS2-2 showed highest N concentration, 65.0 mg N g DW⁻¹, followed by Sinpaldalkong 2 and En1282, and the N content per plant did not show a significant difference between SS2-2 and Sinpaldalkong 2. The N₂ fixation rate was significantly reduced by N top-dressing, but the amount of N₂ fixation was not changed due to an improved dry weight without changes of N concentration. In addition, SS2-2 showed higher NHI, NR and NRE than Sinpaldalkong 2. These results suggested that supernodulating soybean mutants, SS2-2, could be characterized by high N concentration and N₂ fixation regardless of N fertilizer top-dressing due to a higher nitrate tolerance of supernodulating mutants than that of wild-type.     

Effect of Water Regimes and Moisture Stress at Different Growth Stages on Nodule Dynamics,Nitrogenase Activity and Nitrogen Fixation in Soybean (Glycine max (L.) Merrill)*

The effect of water regimes and soil moisture stress on total nodule number, effective nodule number, nodule dry weight, nitrogenase activity and nitrogen fixation in soybean was studied. The effective nodule number, nodule weight, nitrogenase activity and nitrogen fixation increased due to increase in water regimes from 0.4 to 0.6 Cumulative Pan Evaporation (CPE). The stress during either flowering or pod filling reduced the effective nodule number, nodule weight, nitrogenase activity and nitrogen fixation. The amount of nitrogen fixed was maximum with 0.8 CPE (50.09 kg ha^-1 season^-1) followed by 0.6 CPE (42.08 kg ha^-1 season^-1) and 0.4 CPE (33.04 kg ha^-1 season^-1). The amount of nitrogen fixed was reduced by 17.71 % and 26.39 % due to stress during flowering and pod filling respectively compared to normal irrigation.     

甘蓝型油菜重组自交系苗期磷效率的评价

设置低磷P1 (5 μmol L^-1)和高磷P2 (1 000 μmol L^-1)处理水培甘蓝型油菜重组自交系群体的135个株系及亲本的幼苗, 以地上部干重(SDW)、根干重(RDW)、根冠比(R/S)、主根长(AMRL)、地上部磷积累量(SPU)、总磷吸收量(TPU)、磷利用效率(PUE)作为耐性指标, 调查群体各株系和亲本间对缺磷反应的差异, 并对各性状参数与磷吸收、利用效率进行相关性分析。结果表明: (1) 低磷胁迫严重抑制甘蓝型油菜苗期生长, 所调查的各性状均表现出显著差异,其中地上部干重和根干重的变异系数较大;(2) 2种处理条件下, 各株系的地上部干重、根干重、根冠比和主根长4个性状均表现出显著分离, 并呈现正态分布,低磷处理的分离更为明显;(3) 相关性分析表明, 相对地上部干重和根干重可以作为磷效率的主要评价指标,为了避免遗传因素影响, 还应考虑低磷处理下基因型各性状的绝对差异;(4) 通过上述筛选指标,确定065﹑102﹑070为候选的极端磷高效基因型, 105﹑076﹑011等为候选的极端磷低效基因型。     

光合细菌-根瘤菌最佳施用配比的研究

（沈阳农业大学,辽宁沈阳 110161）     

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.     

不同磷效率大豆品种光合特性的比较

以不同磷效率大豆品种为材料, 调查磷高效和磷低效品种的生理指标,测定大豆生育期地上器官干物重,分析光合特性及物质生产特性。结果表明,在低磷条件下, 各生育期的叶绿素含量、可溶性蛋白含量、净光合速率、气孔导度和叶肉导度均以磷高效品种较高, 磷高效品种光合作用较强,CO₂同化能力较高。各生育期, 供试品种的叶绿素含量、可溶性蛋白含量、净光合速率、气孔导度和叶肉导度均在结荚期达到高峰而后下降, 但磷高效品种下降速率低于磷低效品种。磷高效品种的单株和群体均具较高光合效率及生长参数, 并受磷浓度影响较小, 主要原因是其植株吸收磷量较多, 有利于植株体内磷素平衡, 即便在低磷条件下也有相对较强的光合能力,令其在低磷、中磷和高磷条件下都有较高的产量和籽粒磷效率。     

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.