大豆种子低温吸胀相关基因的分离与蛋白变化的研究

大豆 (Glycine max (L.) Meer.)是人们日常生活中不可缺少的食品，也是一种非常重要的油质、蛋白资源。目前根据大豆种子吸胀阶段对低温敏感性的不同，可将其划分成3种生态型：低温非敏感型、低温敏感型及中间型。对于低温非敏感型的种子来讲，4℃下吸胀24小时对其发芽率影响很小，而敏感型种子萌发率不超过5%。我国属于温带大陆性气候，大豆播种后由于温度波动而造成一部分种子不能萌发，最终导致减产甚至绝产的现象普遍存在。高产是育种工作的主要目标，提高逆境胁迫的适应能力是高产的前提和基础，所以从分子角度研究种子吸胀非常必要，一方面能够挖掘新的基因资源，另一方面为今后育种工作提供必要的理论依据。 本试验以此为立足点，低温吸胀非敏感型大豆品种 (Z22)为材料，利用cDNA-AFLP方法及蛋白质技术分离与低温吸胀相关的基因及蛋白，得到结果如下： 第一，本试验成功的分离出4个受低温诱导的基因，半定量RT-PCR方法进一步验证了这4个基因在种子吸胀24小时内受低温诱导。 第二，利用RACE方法成功的得到2个完整的全长基因，在NCBI数据库中查找后发现其中1个基因为新基因，命名为SCHI基因 (SCHI：Soybean chilling-induced gene)。SCHI全长为390bp，编码分子量大约为14.2KD的蛋白；另外一个基因是已知基因，其同源序列已经在其他的物种中得到分离。由于此基因与核糖体蛋白L34高度同源，所以把把这个基因命名为SOL34 (Soybean L34)。 第三，利用半定量RT-PCR方法对基因表达模型进行分析，结果表明：SCHI在种子低温吸胀18～24小时期间诱导表达量最高，而当种子低温吸胀24小时后转入常温下，其表达量在常温下18小时左右迅速下降；ABA (100μM)、PEG (30%，10000)及NaCl (250mM)能够诱导SCHI的表达，在诱导表达量上，ABA和PEG诱导效果最明显，而NaCl���够微弱的诱导此基因表达；对不同生态型的大豆品种而言，低温吸胀过程中，SCHI在非敏感型种子中的表达量高于敏感型种子，但非敏感型和中间型之间没有差别；另外，SCHI在大豆胚轴中是诱导型表达，在叶片和根尖中则是组成型表达。SOL34的表达在萌发前24小时内被低温诱导，但在不同生态型之间没有差别。SOL34在胚轴和根尖中受低温诱导，在叶片中是组成型表达。 第四，SCHI能够在原核生物中表达出相应蛋白，诱导表达蛋白的分子量在26-29KD，大约为理论值的2倍，说明在大肠杆菌中被表达的蛋白以2聚体形式存在。另外低温试验结果表明SCHI能够提高菌落忍耐短时间-20℃低温的能力。 第五，利用双元表达载体把SCHI转入拟南芥植株，经过低温、干旱和盐胁迫后，转基因植株的成活率均高于野生型植株。超表达SOL34的拟南芥植株降低了对低温的耐性；而抑制拟南芥中L34的表达反而提高了植株对低温的抗性。 第六，本试验利用蛋白质等有关试验检测了大豆种子低温吸胀时蛋白质发生的变化。从吸胀 (4℃和22℃下24h)后的大豆胚轴中成功鉴定出上调蛋白点25个，下调蛋白点15个。其中有参与能量代谢反应 (占10%，例如柠檬酸脱氢酶和苹果酸脱氢酶等)、细胞生长与分裂相关反应 (20%，例如LEA蛋白和种子成熟蛋白PM26)、胁迫反应 (10%，如乙醇脱氢酶)、种子宿命和贮藏蛋白 (20%，大豆球蛋白)等蛋白在此过程中发生了变化，暗示种子萌发前期低温吸胀过程中多种代谢发生变化。细胞生长变缓、能量代谢增强、胁迫代谢蛋白的高表达以及贮藏蛋白降解速度减慢等变化都有利于种子在吸胀过程中度过低温环境，为以后的生长作好准备。

低温吸胀对大豆种子呼吸代谢影响的研究

大豆是重要的油料和蛋白植物。在生产实践中，在播种后达到早苗和齐苗是大豆丰收的前提。种子的吸胀冷害是农业生产的严重问题。吸胀冷害发生在种子开始吸水萌动的萌发初始阶段。吸胀冷害不仅发生在高寒地带和低温冷湿地区，尤其在我国东北地区，造成我国乃至全球大豆不同程度的减产。吸胀冷害的原初作用位点在生物膜上，本实验从呼吸代谢的角度研究吸胀冷害对种子活力的影响，探讨吸胀冷害的机制。 本实验选用由黑龙江省黑河农业科学院提供的对低温中度敏感的黑河13 号大豆种子为材料，分别经22°C、10°C 和4°C 恒温培养箱24 h 后，测定其生理指标，通过透射电镜观察细胞超微结构，利用蛋白质组技术研究低温吸胀与种子呼吸代谢的关系，得到的结果如下： 低温吸胀阻碍胚轴膜系统的修复。通过电解质渗漏率测定发现，4°C 到22°C 温度范围内，提高吸胀温度有助于保持细胞膜的完整性，显著降低吸胀后胚轴电解质渗漏。在低温下吸胀，胚轴活性氧清除酶的活性受到抑制，活性氧含量增加，增强了膜脂过氧化作用，进而导致种子活力下降。 通过透射电子显微镜观察，大豆种子在22°C 吸胀24 h 后，胚轴细胞液泡明显变大，在细胞中所占比例很高，并且细胞内膜系统比较发达，能清晰观察到细胞核，线粒体，质膜，内质网整齐有序的形状。胚轴的细胞含有其它结构正常的细胞器，包括细胞壁，胞间连丝，淀粉粒和油体等。线粒体的外膜、内膜、嵴发育较完善。10°C 和4°C 的吸胀严重损伤了胚轴中细胞器的修复，细胞膜不规则，没有发现内质网和胞间连丝，线粒体的体积较小以及膜系统不发达，尤其是4°C 吸胀的胚轴中细胞器的损伤更加严重，细胞膜系统紊乱。 低温吸胀抑制了线粒体从轻线粒体向重线粒体的修复，以及线粒体的耗氧能力。22°C 吸胀的线粒体的总体耗氧能力较高，电子传递主要是利用复合体I 的电子传递途径。10°C 吸胀的线粒体总体耗氧呼吸较低，且其线粒体的电子传递主要以复合体II 的途径。4°C 吸胀的线粒体的耗氧能力则更低。 将分离得到的线粒体进行的蛋白质组分析，共分离400 多个蛋白点，其中有20 个点有表达差异。经ESI-Q-TOF-MS/MS 鉴定，六个下调的蛋白质分别为ATP 合成酶的亚基 (线粒体的氧化磷酸化)，线粒体延长因子Tu（线粒体基因组转录）， 苹果酸脱氢酶（三羧酸循环），精氨酸酶（尿素循环）和2 个线粒体chaperonin-60 （热稳定蛋白）。这些蛋白在低温吸胀时下调表达，影响了线粒体的正常生理代谢，说明它们在维持线粒体正常代谢中起到了重要的作用。 综上所述，低温吸胀影响了线粒体的结构和生理功能的修复，减少了能量和中间物质供应给种子萌发，造成了种子活力的下降。

黄土丘陵区免耕条件下两种土壤酶活性变化

为了确定合理环保的耕作制度,2007~2008安塞田间定位试验黄土丘陵旱作农区大豆(Glycine maxL)、玉米(Zea maysL)、红小豆(Semen Phaseoli)、马铃薯(Solanum tuberosumLinn.)在翻耕化肥(CF)、翻耕有机肥(CM)、翻耕无肥(CN)、免耕化肥(NF)、免耕有机肥(NM)、免耕无肥(NN)等水平下的农田土壤脲酶、蔗糖酶活性。结果表明:在作物花期,大豆、玉米土壤脲酶活性较高,蔗糖酶活性较低,而红小豆、马铃薯则与之相反,差异极显著。到作物收获后,玉米土壤脲酶、蔗糖酶活性增高,增幅在83%以上,而马铃薯、红小豆、大豆三种作物土壤脲酶活性降低,降幅在10%以上,蔗糖酶活性增强,增幅在40%以上。从花期到收获后,免耕降低了土壤脲酶活性,提高了土壤蔗糖酶活性,在黄土丘陵沟壑旱作农区两种土壤酶活性表现较优的作物为大豆、玉米,较优的处理为NM玉米、NF大豆,其田间环保效应为:作物生长期间两种酶活性比较高,收获后两种酶活性则降低,有利于提高作物生长期土壤肥效利用率,减少作物收获后温室气体的排放。

不同类型作物对干湿交替环境的反应

通过对干湿交替环境下春小麦、马铃薯、大豆和玉米等作物的产量、水分利用效率及光合作用、蒸腾作用、气孔导度等生理变化的研究表明 :( 1 )春小麦和马铃薯在干湿交替环境下可获得与充分供水相当的产量而它们的水分利用效率却显著提高 ,大豆减产幅度较大 ,玉米减产严重 ,其水分利用效率显著低于全湿处理 ;( 2 )浇水后各作物的光合速率、蒸腾速率和气孔导度都有所增加 ,但不同作物增加的幅度不同 ,就是同一作物各指标的增幅也不同 ;( 3)干湿交替环境下同化物的运输模式有利于春小麦籽粒的充实和马铃薯块茎的膨大 ,而不利于玉米产量的形成 ;( 4 )产量不仅决定于营养生长阶段 ,更主要决定于生殖生长阶段。此外 ,还就干湿交替过程中若干生理变化和经济产量形成机制作了初步探讨。

Classificacao taxonomica, baseada na caracterizacao molecular, das estirpes de rizobio recomendadas para a culturas da soja e do feijoeiro.

As culturas da soja [Glycine max (L.) Merril] e do feijoeiro (Phaseolus vulgaris) sao de grande importancia economica e social para o Brasil e ambas podem ser capazes de suprir suas necessidades do nutriente nitrogenio pela simbiose com bacterias da familia Rhizobiaceae. Para garantir a maximizacao do processo biologico, porem, deve-se proceder a inoculacao das sementes com estirpes de rizobio eficientes e competitivas, recomendadas pela pesquisa. No Brasil, sao comercializadas, anualmente, cerca de 13 milhoes de doses de inoculantes, sendo 99% destinadas a cultura da soja. Neste trabalho, determinou-se a posicao taxonomica das estirpes de rizobio recomendadas comercialmente para as duas culturas por duas tecnicas, o RFLP-PCR da regiao correspondente ao 16S rRNA (regiao conservada entre bacterias mas suficientemente variavel e carregando informacoes que permitem a determinacao das relacoes filogeneticas entre bacterias) e o sequenciamento parcial dos genes desta regiao. O sequenciamento parcial permitiu definir que duas das estirpes recomendadas para a cultura da soja, SEMIA 587 e SEMIA 5019, pertencem a especie Bradyrhizobium elkanii e as duas outras, SEMIA 5079 e SEMIA 5080, a especie B. japonicum. As duas estirpes recomendadas para a cultura do feijoeiro, SEMIA 4077 e SEMIA 4080, pertencem a especie Rhizobium tropici. As sequencias obtidas para essas estirpes foram depositadas no banco mundial de genes.

Mancha Ojo de Rana (Cercospora sojina K. Hara) y su efecto sobre rendimiento y componentes industriales del grano de soja : proteína y aceite

La mancha ojo de rana (MOR), causada por Cercospora sojina K. Hara, se encuentra dentro del complejo de enfermedades fúngicas de fin de ciclo (EFC), que afectan al cultivo de soja (Glycine max (L.) Merr.) preferentemente en los estadíos reproductivos. Es una enfermedad común en la soja en la mayor parte de las regiones sojeras del mundo, incluyendo Argentina. Con los objetivos de (i) estudiar el efecto de MOR sobre rendimiento y calidad industrial del grano de soja y (ii) analizar la respuesta a distintos momentos de aplicación de estrobilurina+triazol durante el ciclo reproductivo del cultivo para disminuir el daño de esta enfermedad evitando pérdidas del rendimiento y de calidad del grano, se realizó un ensayo durante la campaña agrícola 2010-2011, en la localidad de Oncativo (Cordoba), que involucró 2 genotipos contrastantes en susceptibilidad a MOR. Se realizaron para cada cultivar cuatro tratamientos con una mezcla de triazol y estrobilurina: T1= aplicación en R3; T2= aplicación en R3+R5; T3= testigo enfermo (sin aplicación de fungicida) y T4= testigo sano (aplicacion cada 20 dias a partir de R1). Se determinó el nivel de incidencia y severidad de la enfermedad, rendimiento, número de grano por m2 (NGm2), peso de mil semillas (PMA), concentración de proteína (Pr) y aceite (Ac) del grano. Se observaron distintos niveles de severidad de MOR entre los tratamientos del cultivar DM3700. Todos los tratamientos que involucraron a DM3810 mostraron hojas sin manchas de MOR. Se encontró una correlación negativa entre la severidad de MOR y el rendimiento alcanzado por el cultivo. Los valores más elevados de severidad (37,3 por ciento de manchas foliares) se correspondieron con los rendimientos mas bajos (2117 kg ha-1) en el cultivar DM3700. Todos los tratamientos aplicados a DM3810 exhibieron los rendimientos mas elevados y no se diferenciaron estadisticamente entre si (3478 kg ha-1). DM3700 alcanzó rendimientos similares a DM3810 sólo cuando se aplicó la mezcla de fungicidas en T1 y T4, no diferenciándose entre si ambos tratamientos y mostrando un incremento en el rendimiento de aproximadamente 35,5 por ciento comparado con su testigo enfermo. Ambos componentes del rendimiento disminuyeron en el testigo enfermo DM3700; sin embargo, las reducciones en el NGm2 fueron más pronunciadas que las del PMAj y; por lo tanto, disminuciones en el NGm2 parecían ser el principal componente de la pérdida de rendimiento. No se observaron incrementos de rendimiento, NGm2 y/o PMAj atribuídos a la mezcla estrobilurinas + triazoles en los tratamientos que involucraron a DM3810 ante ausencia de MOR. La severidad máxima alcanzada por MOR en el genotipo mas susceptible (37,3 por ciento) de area foliar dañada en el tratamiento enfermo de DM3700) no fue suficiente como para causar disminución significativa en la acumulación del aceite. Según nuestro conocimiento este trabajo es el primero en demostrar que genotipos susceptibles a MOR disminuyen su rendimiento, pero no la calidad química del grano, si la severidad es . 37,3 por cientoy pueden ser controlados con una única aplicación de estrobilurina+triazol en R3. Concluyéndose que la elección de genotipos poco susceptibles a MOR (como DM3810) es efectiva para controlar C. sojina, sin beneficios adicionales en el rendimiento debidos a la aplicacion de fungicidas.

Interacción entre fecha y densidad de siembra en cultivos invernales

El cultivo de soja (Glycine max L.) de primera predomina en las secuencias agrícolas de la provincia de Entre Ríos, desperdiciándose una enorme cantidad de recursos al realizar un sólo cultivo por año. El estudio comparado de la interacción entre fecha de siembra y densidad en cultivos invernales ayudaría a identificar la mejor combinación para maximizar el rendimiento, alentando la intensificación de la secuencia de producción. El objetivo de este trabajo fue estudiar de manera comparada la interacción entre la fecha de siembra y la densidad de siembra en arveja proteica (Pisum sp.), lino (Linum usitatissimum L.), trigo ciclo corto y ciclo largo (Triticum aestivum L.). El experimento fue conducido en la EEA Paraná del INTA durante la campaña 2008/09, se evaluaron tres fechas de siembra (FS) y los tratamientos consistieron en una combinación factorial de cultivos invernales y densidades de siembra (DS). Se determinó la biomasa total acumulada hasta madurez, el rendimiento, los componentes y subcomponentes del rendimiento. Además, en la fecha de siembra más temprana se cuantificaron los componentes ecofisiólogicos condicionantes de la acumulación de biomasa, i.e. intercepción de la radiación fotosinteticamente activa (RFA), eficiencia en el uso de la radiación (EUR) y de partición. La eficiencia de intercepción de RFA como así también la EUR difirió significativamente en respuesta a la densidad de siembra, pero no fue significativo su efecto sobre la RFA acumulada y la evolución de la biomasa durante el ciclo de los cultivos. En ninguno de los cultivos estudiados existió interacción significativa FSxDS sobre el rendimiento o sus componentes numéricos, sólo en los cultivares de trigo se registraron efectos significativos de la interacción FSxDS sobre los subcomponentes del rendimiento. Para estudiar mejor la interacción FSxDS en especies con diferente plasticidad sería conveniente explorar su comportamiento sin limitaciones hídricas, complementando dichos estudios con la medición de variables ecofisol��gicas

Evaluación del balance de energía en un cultivo de soja bajo condiciones hídricas extremas

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Utilización de un residuo mineral de baja ley en fósforo como fertilizante

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Rapid [Gamma]-amino acid synthesis and the inhibition of the growth and development of oblique banded leaf-roller larvae

The hypothesis that rapid y-aminobutyric acid (GABA) accumulation is a plant defense against phytophagous insects was investigated. Simulation of mechanical damage resulting from phytophagous insect activity increased soybean (Glycine max L.) leaf GABA 10- to 25-fold within 1 to 4 min. Pulverizing leaf tissue resulted in a value of 2. 15 (±O. 11 SE) ~mol GABA per gram fresh weight. Increasing the GABA levels in a synthetic diet from 1.6 to 2.6 Jlffiol GABA per gram fresh weight reduced the growth rates, developmental rates, total biomass (50% reduction), and survival rates (30% reduction) of cultured Oblique banded leaf-roller (OBLR) (Choristonellra rosacealla Harris) larvae. In field experiments OBLR larvae were found predominantly on young terminal leaves which have a reduced capacity to produce GABA in response to mechanical damage. Glutamate decarboxylase (GAD) is a cytosolic enzyme which catalyses the decarboxylation of L-Glu to GABA. GAD is a calmodulin binding enzyme whose activity is stimulated dramatically by increased cytosolic H+ or Ca2 + ion concentrations. Phytophagous insect activity will disrupt the cellular compartmentation of H+ and Ca2 +, activate GAD and subsequent GABA accumulation. In animals GABA is a major inhibitory neurotransmitter. The possible mechanisms resulting in GABA inhibited growth and development of insects are discussed.

Effects of row spacing on productivity and nodulation of two soybean varieties under hot sub-moist tropical conditions in south-western Ethiopia

The objective of this study was to determine the optimum row spacing to improve the productivity of two soybean (Glycine max L.) varieties under the tropical hot sub-moist agroecological conditions of Ethiopia. A two-year split-plot design experiment was conducted to determine the effect of variety (Awasa-95 [early-maturing], Afgat [medium-maturing]) and row spacing (RS: 20, 25, 30, 35, 40, 45, 50, 55, 60 cm) on the productivity, nodulation and weed infestation of soybean. Seed and total dry matter (TDM) yield per ha and per plant, and weed dry biomass per m^2 were significantly affected by RS. Soybean variety had a significant effect on plant density at harvest and some yield components (plant height, number of seeds/pod, and 1000 seed weight). Generally, seed and TDM yield per ha and per plant were high at 40 cm RS, and weed dry biomass per m^2 was higher for RS >= 40 cm than for narrower RS. However, the results did not demonstrate a consistent pattern along the RS gradient. The medium-maturing variety Afgat experienced higher mortality and ended up with lower final plant density at harvest, but higher plant height, number of seeds per pod and 1000 seed weight than the early-maturing variety Awasa-95. The results indicate that 40 cm RS with 5 cm plant spacing within a row can be used for high productivity and low weed infestation of both soybean varieties in the hot sub-moist tropical environment of south-western Ethiopia.

Effects of fully open-air CO2 elevation on leaf ultrastructure, photosynthesis and yield of two soybean cultivars.

The objective of this study was to investigate the effect of elevated (550 ± 17 μmol mol���1) CO2 concentration ([CO2]) on leaf ultrastructure, leaf photosynthesis and seed yield of two soybean cultivars [Glycine max (L.) Merr. cv. Zhonghuang 13 and cv. Zhonghuang 35] at the Free-Air Carbon dioxide Enrichment (FACE) experimental facility in North China. Photosynthetic acclimation occurred in soybean plants exposed to long-term elevated [CO2] and varied with cultivars and developmental stages. Photosynthetic acclimation occurred at the beginning bloom (R1) stage for both cultivars, but at the beginning seed (R5) stage only for Zhonghuang 13. No photosynthetic acclimation occurred at the beginning pod (R3) stage for either cultivar. Elevated [CO2] increased the number and size of starch grains in chloroplasts of the two cultivars. Soybean leaf senescence was accelerated under elevated [CO2], determined by unclear chloroplast membrane and blurred grana layer at the beginning bloom (R1) stage. The different photosynthesis response to elevated [CO2] between cultivars at the beginning seed (R5) contributed to the yield difference under elevated [CO2]. Elevated [CO2] significantly increased the yield of Zhonghuang 35 by 26% with the increased pod number of 31%, but not for Zhonghuang 13 without changes of pod number. We conclude that the occurrence of photosynthetic acclimation at the beginning seed (R5) stage for Zhonghuang 13 restricted the development of extra C sink under elevated [CO2], thereby limiting the response to elevated [CO2] for the seed yield of this cultivar.

The pgip family in soybean and three other legume species: evidence for a birth-and-death model of evolution

Background Polygalacturonase-inhibiting proteins (PGIPs) are leucine-rich repeat (LRR) plant cell wall glycoproteins involved in plant immunity. They are typically encoded by gene families with a small number of gene copies whose evolutionary origin has been poorly investigated. Here we report the complete characterization of the full complement of the pgip family in soybean (Glycine max [L.] Merr.) and the characterization of the genomic region surrounding the pgip family in four legume species. Results BAC clone and genome sequence analyses showed that the soybean genome contains two pgip loci. Each locus is composed of three clustered genes that are induced following infection with the fungal pathogen Sclerotinia sclerotiorum (Lib.) de Bary, and remnant sequences of pgip genes. The analyzed homeologous soybean genomic regions (about 126 Kb) that include the pgip loci are strongly conserved and this conservation extends also to the genomes of the legume species Phaseolus vulgaris L., Medicago truncatula Gaertn. and Cicer arietinum L., each containing a single pgip locus. Maximum likelihood-based gene trees suggest that the genes within the pgip clusters have independently undergone tandem duplication in each species. Conclusions The paleopolyploid soybean genome contains two pgip loci comprised in large and highly conserved duplicated regions, which are also conserved in bean, M. truncatula and C. arietinum. The genomic features of these legume pgip families suggest that the forces driving the evolution of pgip genes follow the birth-and-death model, similar to that proposed for the evolution of resistance (R) genes of NBS-LRR-type.

Storage proteins and cell wall mobilisation in seeds of Sesbania virgata (Cav.) Pers. (Leguminosae)

The endosperm of seeds of Sesbania virgata (Cav.) Pers. accumulates galactomannan as a cell wall storage polysaccharide. It is hydrolysed by three enzymes, one of them being alpha-galactosidase. A great amount of protein bodies is found in the cytoplasm of endospermic cells, which are thought to play the major role as a nitrogen reserve in this seed. The present work aimed at understanding how the production of enzymes that degrade storage compounds is controlled. We performed experiments with addition of inhibitors of transcription (actinomycin-d and alpha-amanitin) and translation (cycloheximide) during and after germination. In order to follow the performance of storage mobilisation, we measured fresh mass, protein contents and alpha-galactosidase activity. All the inhibitors tested had little effect on seed germination and seedling development. Actinomycin-d and cycloheximide provoked a slight inhibition of the storage protein degradation and concomitantly lead to an elevation of the alpha-galactosidase activity. Although alpha-amanitin showed some effect on seedling development at latter stages, it presented the former effect and did not change galactomannan degradation performance. Our data suggest that some of the proteases may be synthesised de novo, whereas alpha-galactosidase seems to be present in the endosperm cells probably as an inactive polypeptide in the protein bodies, being probably activated by proteolysis when the latter organelle is disassembled. These evidences suggest the existence of a connection between storage proteins and carbohydrates mobilisation in seeds of S. virgata, which would play a role by assuring a balanced afflux of the carbon and nitrogen to the seedling development.

Relative roles of glyceollin, lignin and the hypersensitive response and the influence of ABA in compatible and incompatible interactions of soybeans with Phytophthora sojae

The relative roles of glyceollin, lignin and the hypersensitive response (HR) in pathogen containment and restriction were investigated in soybean (Glycine max L. [Merr.]) cultivars that were inoculated with Phytophthora sojae Kaufmann and Gerdemann. Concentrations of endogenous abscisic acid (ABA) levels in etiolated soybean hypocotyls were reduced by application of the ABA biosynthesis inhibitor norflurazon or raised by exogenous ABA application. Incompatible interactions in leaves and hypocotyls were characterized by HR, phenolic and lignin deposition and glyceollin accumulation. Compatible interactions resulted in light coloured, water-soaked spreading lesions with minimal lignin deposition or glyceollin accumulation and the absence of an HR. Norflurazon treatment restricted the spread of the pathogen and increased glyceollin accumulation in compatible tissues. Exogenous ABA addition caused spreading lesions in normally incompatible interactions and reduced glyceollin accumulation. Phenolic deposition and HR were unchanged by either treatment in incompatible or compatible interactions. The uncoupling of glyceollin synthesis from the HR and phenolic and lignin deposition by ABA and norflurazon treatment showed that glyceollin is a major factor in restriction of the pathogen during these interactions.