The seed lectins of black locust (robinia pseudoacacia) are encoded by two genes which differ from the bark lectin genes

Two lectins were isolated from Robinia pseudoacacia (black locust) seeds using affinity chromatography on fetuin-agarose, and ion exchange chromatography on a Neobar CS column. The first lectin, R. pseudoacacia seed agglutinin I, referred to as RPsAI, is a homotetramer of four 34 kDa subunits whereas the second lectin, referred to as RPsAII, is composed of four 29 kDa polypeptides. cDNA clones encoding the polypeptides of RPsAI and RPsAII were isolated and their sequences were determined. Both polypeptides are translated from mRNAs of ca. 1.2 kb encoding a precursor carrying a signal peptide. Alignment of the deduced amino acid sequences of the different clones indicates that the 34 and 29 kDa seed lectin polypeptides show 95% sequence identity. In spite of this striking homology, the 29 kDa polypeptide has only one putative glycosylation site whereas the 34 kDa subunit has four of these sites. Carbohydrate analysis revealed that the 34 kDa possesses three carbohydrate chains whereas the 29 kDa polypeptide is only partially glycosylated at one site. A comparison of the deduced amino acid sequences of the two seed and three bark lectin polypeptides demonstrated unambiguously that they are encoded by different genes. This implies that five different genes are involved in the control of the expression of the lectins in black locust.Abbreviations LECRPAs cDNA clone encoding Robinia pseudoacacia seed lectin - LoLI Lathyrus ochrus isolectin I - PsA Pisum sativum agglutinin - RPbAI Robinia pseudoacacia bark agglutinin I - RPbAII Robinia pseudoacacia bark agglutinin II - RPsAI Robinia pseudoacacia seed agglutinin I - RPsAII Robinia pseudoacacia seed agglutinin II     

Formation of heartwood substances in the stem of Robinia pseudoacacia L.

Summary The activities of two key enzymes in flavonoid biosynthesis, phenylalanine ammonia-lyase (PAL, E.C. 4.3.1.5) and chalcone synthase (CHS, E.C. 2.3.1.74) were determined in the trunkwood of Robinia pseudoacacia L. The trees under investigation were cut at different times of the year (September, November, January and April). At all times PAL is active, both in the youngest wood layer (the outermost growth ring) and at the sapwood heartwood boundary. On the other hand, CHS is active exclusively in the vicinity of the heartwood boundary. The results indicate that PAL is involved both in the formation of lignin (outermost annual ring), and in flavonoid biosynthesis (heartwood boundary). Highest activity of both PAL and CHS could be measured at the sapwood heartwood boundary in the tree felled in November, indicating that heartwood formation was occurring mainly at that time. The flavonoids accumulated in the heartwood are obviously formed in situ and seem to be transported only to a minor extent — if at all — via the phloem and the ray cells to the heartwood.     

Microautoradiographic localisation of [3H]sucrose and [3H]mannitol in Robinia pseudoacacia pulvinar tissues during phytochrome-mediated nyctinastic closure

Summary. We have analysed the incorporation of [³H]sucrose and [³H]mannitol in pulvinar motor cells of Robinia pseudoacacia L. during phytochrome-mediated nyctinastic closure. Pairs of leaflets, excised 2 h after the beginning of the photoperiod, were fed with 50 mM [³H]sucrose or [³H]mannitol, irradiated with red (15 min) or far-red (5 min) light and placed in the dark for 2–3 h. Label uptake was measured in whole pulvini by liquid scintillation counting. The distribution of labelling in pulvinar sections was assessed by both light and electron microautoradiography. [³H]Sucrose uptake was twice that of [³H]mannitol incorporation in both red- and far-red-irradiated pulvini. In the autoradiographs, [³H]sucrose and [³H]mannitol labelling was localised in the area from the vascular bundle to the epidermis, mainly in vacuoles, cytoplasm, and cell walls. Extensor and flexor protoplasts displayed a different distribution of [³H]sucrose after red and far-red irradiation. Far-red light drastically reduced the [³H]sucrose incorporation in extensor protoplasts and caused a slight increase in internal flexor protoplasts. After red light treatment, no differences in [³H]sucrose labelling were found between extensor and flexor protoplasts. Our results indicate a phytochrome control of sucrose distribution in cortical motor cells and seem to rule out the possibility of sucrose acting as an osmoticum. Correspondence and reprints: Unidad de Fisiología Vegetal, Facultad de Biología, Universidad de Barcelona, Avenida Diagonal 645, 08028 Barcelona, Spain.     

Genetic relatedness in open-pollinated families of two leguminous tree species,Robinia pseudoacacia L. and Gleditsia triacanthos L.

Summary When conducting tree breeding experiments, geneticists often assume that individuals from open-pollinated families are halfsibs. The reliability of this assumption was tested using data from enzyme electrophoresis to estimate the genetic relatedness among progeny within 22 open-pollinated families of Robinia pseudoacacia L. (black locust) and 34 open-pollinated families of Gleditsia triacanthos L. (honey locust) from natural stands. An algorithm employing population estimates of fixation indices, pollen allele frequencies, and selfing rates was used to calculate the mean expected number of alleles in common across loci under assumptions of either full-sib (i.e., a single pollen parent) or half-sib (i.e., random mating) relationships. For each open-pollinated family, the average coefficient of relationship among progeny was calculated by linear interpolation from the observed number of alleles in common. For most families of both species, coefficients were significantly higher than 0.25 (half-sib relation), but were significantly lower than 0.50 (full-sib relation). These results suggest that the assumption of a half-sib relationship among progeny of open-pollinated families is violated for these tree species. More critical to the estimation of heritabilities and the prediction of genetic gains was the observation that estimates of the coefficient of relationship varied widely among open-pollinated families (for R. pseudoacacia r ₀=0.20–0.43, mean=0.34; for G. triacanthos r ₀=0.29–0.55, mean=0.36).     

土壤水分和氮素有效性对刺槐幼苗叶片活性氧产生和清除的影响

采用施加氮肥和人工控水的方法,以一年生刺槐幼苗为材料进行盆栽实验,探讨提高土壤氮素含量对不同土壤水分条件下刺槐幼苗叶片中活性氧产生和清除的影响。结果表明:(1)相同氮素水平下,降低土壤水分含量引起刺槐生物量和叶片光合色素含量降低,而过氧化氢(H2O2)含量升高;抗氧化酶系统中的超氧化物歧化酶(SOD)和过氧化物酶(POD)活性和过氧化氢酶(CAT)活性不同程度降低;抗氧化剂中抗坏血酸(ASA)含量和还原型谷胱甘肽(GSH)含量均有所提高;MDA含量逐渐升高,而同期细胞膜相对电导率显著升高。(2)相同水分条件下,提高土壤氮素水平显著提高了刺槐幼苗叶片光合色素含量,同时也一定程度提高了总生物量,显著降低了H2O2含量;SOD、POD和CAT活性不同程度升高;ASA含量和GSH含量则表现出不同程度下降;相对电导率显著降低同时MDA含量一定程度降低。因此,增加土壤氮素有效性可显著提高刺槐幼苗叶片光合色素含量,显著抑制活性氧的产生,一定程度提高总生物量和抗氧化酶活性,降低膜脂过氧化程度,从而有利于缓解干旱引起的伤害。     

Lipids and lipolytic enzymes in the trunkwood ofRobinia pseudoacacia L. during heartwood formation

The radial distribution of membrane and storage lipids was determined in the trunkwood ofRobinia pseudoacacia L. The trees were felled in November at the time of heartwood formation and fluctuations in the amount and composition of phospholipids, free sterols, steryl esters, diand triacylglycerols, and free fatty acids were investigated across the sapwood-heartwood boundary. The individual compounds were identified and quantified by thin layer chromatography, enzymatic and colorimetric assays, and by capillary gas chromatography. Phospholipids show a significant decrease towards the boundary area, and in the heartwood only trace amounts can be detected. The same pattern is observed for free sterols in the sapwood; in the heartwood, however, they reach maximum values with increasing depth of the trunk. Steryl esters exhibit a complementary behaviour by accumulating at the periphery of the heartwood. No concentration changes are found in the total amounts of diacylglycerols and free fatty acids. In contrast, the triacylglycerol concentration declines steadily across the trunk. With regard to qualitative composition, free fatty acids and the fatty acid moieties of the esterified constituents vary in their chain length from 14 to 24 carbon atoms and have up to three double bonds. A radial gradient in the ratio saturated/unsaturated fatty acids can be observed: except for the phospholipid fraction the relative amounts of unsaturated fatty acids increase in centripetal direction. Seven phospholipids were identified: phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, diphosphatidylglycerol, and phosphatidic acid, which constitutes the major proportion. In the sterol group, sitosterol is the most abundant component. The composition of the esterified sterols remains constant across the trunk cross-section, whereas the relative frequencies of individual free sterols change markedly.     

Phenotypic,genomic and phylogenetic characteristics of rhizobia isolated from root nodules of <Emphasis Type="Italic">Robinia pseudoacacia</Emphasis> (black locust) growing in Poland and Japan

Rhizobial strains, rescued from the root nodules of Robinia pseudoacacia growing in Japan and Poland, were characterized for the phenotypic properties, genomic diversity as well as phylogeny and compared with the reference strains representing different species and genera of nodule bacteria. They had a moderately slow growth rate, a low tolerance to antibiotics, a moderate resistance to NaCl and produced acid in yeast mannitol agar. Cluster analysis based on the phenotypic features divided all bacteria involved in this study into four phena, comprising: (1) Rhizobium sp. + Sinorhizobium sp., (2) Bradyrhizobium sp., (3) R. pseudoacacia microsymbionts + Mesorhizobium sp., and (4) Rhizobium galegae strains at similarity coefficient of 74%. R. pseudoacacia nodule isolates and Mesorhizobium species were placed on a single branch clearly distinct from other rhizobium genera lineages. Strains representing R. pseudoacacia microsymbionts shared 98–99% 16S rDNA sequence identity with Mesorhizobium species and in 16S rDNA phylogenetic tree all these bacteria formed common cluster. The rhizobia tested are genomically heterogeneous as indicated by the AFLP (Amplified Fragment Length Polymorphism) method. The bacteria studied exhibited high degree of specificity for nodulation. Nitrogenase structural genes in these strains were located on 771–961 kb megaplasmids. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Cloning of a lectin cDNA and seasonal changes in levels of the lectin and its mRNA in the inner bark ofRobinia pseudoacacia

A cDNA clone encoding a lectin was isolated by immunological screening of an expression library prepared from poly(A)⁺ RNA from the inner bark ofRobinia pseudoacacia. The cDNA clone (RBL104) had an open reading frame of 858 bp that encoded a polypeptide with a predicted molecular weight of 31210. This molecular weight corresponded closely to that of a polypeptide immunoprecipitated from products of translationin vitro of the poly(A)⁺ RNA. Thus, RBL104 appeared to be a full-length cDNA. The N-terminal amino acid sequence of the purified lectin protein matched a portion of the predicted amino acid sequence. It appeared that the lectin was synthesized as a precursor that consisted of a putative signal peptide of 31 amino acids and a mature polypeptide of 255 amino acids. Southern blot analysis of the genomic DNA revealed that the lectin was encoded by a small multigene family. The lectin was mostly localized in the axial and ray parenchymal cells of the inner bark. A small amount of lectin was also found in the axial and ray parenchymal cells of the xylem. The lectin accumulated in the inner bark in September, remained at high levels during the winter and disappeared in May. The mRNA for the lectin was detected from August to the following March. The appearance and disappearance of the mRNA were observed prior to those of the lectin protein.     

Effects of Robinia pseudoacacia on the undergrowth of herbaceous plants and soil properties in the Loess Plateau of China

刺槐对黄土高原林下草本植物及土壤性质的影响 外来物种刺槐(Robinia pseudoacacia)的引入会对林下草本植物、土壤性质及二者之间的相互关系产 生一定影响,且这种影响可能与植被带有关。然而,在黄土高原较大面积范围内,关于刺槐对林下草本植物和土壤影响的研究依然很少。我们以黄土高原草原带、森林草原带和森林带作为研究区,选择了两种冠层植物类型：刺槐林分和相邻的原生植被。我们测定了5个叶片功能性状：叶片碳含量、叶片氮含量、叶片磷含量、比叶面积和叶组织密度,并计算了群落功能多样性、物种多样性和群落加权性状值。研究结果表明,(1)与原生植被相比,3个植被带内林下草本植物群落的叶氮、叶磷和比叶面积的群落加权性状值均显著提高,而叶碳和叶组织密度的群落加权性状值显著降低。(2)物种多样性、功能多样性和群落生物量在草原带内呈下降趋势,在森林带内呈上升趋势,在森林草原带内差异不显著。(3)与原生植被相比,土壤有机碳和土壤全氮仅在森林带内显著减少。(4)林下草本植物群落特性与土壤之间的关系是受刺槐和植被带所依赖的。总的来说,刺槐对林下草本植物和土壤性质的影响与植被带密切相关。该研究结果对黄土高原人工植被恢复与重建的规划具有重要指导意义。     

Tardiphaga robiniae gen. nov., sp. nov., a new genus in the family Bradyrhizobiaceae isolated from Robinia pseudoacacia in Flanders (Belgium)

Gram-negative, rod-shaped bacteria were isolated from Robinia pseudoacacia root nodules. On the basis of the 16S rRNA gene phylogeny, they are closely related to Bradyrhizobium, Rhodopseudomonas and Nitrobacter species (97% sequence similarity), belonging to the class Alphaproteobacteria and family Bradyrhizobiaceae. The results of physiological and biochemical tests together with sequence analysis of housekeeping genes (atpD, dnaK, gyrB, recA and rpoB) allowed differentiation of this group from other validly published Bradyrhizobiaceae genera. NodA, nodC and nifH genes could not be amplified. On the basis of genotypic and phenotypic data, these organisms represent a novel genus and species for which the name Tardiphaga robiniae gen. nov., sp. nov. (LMG 26467(T)=CCUG 61473(T)), is proposed.     

黄土丘陵区刺槐与油松人工林生态系统生态化学计量特征

为阐明不同人工林生态系统间生态化学计量特征的差异,采用野外采样与室内分析相结合的方式分析了陕北黄土丘陵区落叶阔叶树种刺槐和常绿针叶树种油松人工林乔木、灌草、枯落物和土壤(土层深度0—100cm)C、N、P化学计量特征。结果表明:1)刺槐乔木各器官(叶、枝、干、皮、根)C含量显著低于油松,但N和P含量显著高于油松。因此,油松的C∶N和C∶P显著大于刺槐,而N∶P小于刺槐。2)刺槐林下枯落物N和P含量显著高于油松,但C含量显著小于油松。此外,油松林下枯落物C∶N(70.21)大于刺槐林下枯落物C∶N(19.71),说明油松林下枯落物分解较慢,有利于养分的存储。3)刺槐和油松人工林土壤C、N含量均随土壤深度增加而减少,P含量则基本保持不变。刺槐人工林土壤中C含量低于油松,N、P含量在两者之间无显著差异。4)刺槐人工林内乔灌草叶、枯落物与土壤C、N、P及其计量比的相关性多集中在10—20、20—30cm土层,而油松林中各组分与土壤营养元素的相关性相对较小,其中20—30cm土层中无显著相关性,说明相比刺槐人工林而言,油松人工林内土壤层N、P供应量对植物叶片N、P含量影响不显著。本研究为深入了解黄土丘陵区生态系统养分耦合循环机制奠定了基础,同时也为黄土丘陵区的植被恢复工作提供了一定的指导意义。     

黄土高原南北样带刺槐林土壤碳、氮、磷生态化学计量特征

为了解黄土高原南北样带刺槐林土壤碳（C）、氮（N）、磷（P）化学计量特征,采集了黄土高原南北样带上12个典型样点的刺槐林土壤,测定了土壤有机碳（SOC）、全氮（TN）、全磷（TP）含量并分析其生态化学计量特征。结果表明：刺槐林土壤SOC、TN、TP含量均随降水量降低而降低,且土壤SOC与TN的空间变化具有一致性;土壤SOC、TN、TP及其生态化学计量比与降雨量呈显著正相关（P < 0.05）,且TN、C：P、N：P与土壤含水量也呈显著正相关（P < 0.05）。     

Facilitation and Inhibition of Seedlings of an Invasive Tree (Acer platanoides) by Different Tree Species in a Mountain Ecosystem

Facilitation is known to be an important process structuring natural plant communities. However, much less is known about its role in facilitating the invasion of ecosystems by non-native plant species. In this study we evaluated the effects of invasive (Acer platanoides) and native (Pseudotsuga menziesii) forest types on the performance of A. platanoides seedlings, and related these effects to structural and functional properties associated with the two forest types, in a native P. menziesii forest that is being invaded by A. platanoides. Acer platanoidesseedlings had higher densities, recruitment, and survival, and experienced less photoinhibition and water stress when beneath conspecific canopies than in the adjacent P. menziesii forest. Soil moisture and canopy cover were greater in the invaded patch than the native forest. There was no difference in soil fertility or understory light levels between locations. These demographic (i.e. seedling survival), physiological, and environmental differences appeared to be due to the effects of A. platanoides and P. menziesii trees. Thus, Acer trees appear to produce a more mesic environment by modifying the structure and phenology of the forest canopy and by altering the timing of transpirational water loss relative to P. menziesii. Environmental modification by invaders that lead to positive effects on conspecifics may help us to understand the dramatic success and lag periods of some invasive species     

黄土高原中西部刺槐人工林生态系统碳密度及其影响因子

为阐明黄土高原中西部刺槐人工林碳密度区域分布特征及其主要影响因子,基于野外样地调查和室内样品分析估算了黄土高原中西部4个栽培区域的刺槐人工林生态系统碳密度及其分布特征,并利用相关性分析和主成分分析分析了影响生态系统碳密度的主要因子(林分、地形、土壤和气候等)。结果表明:调查区5个林龄的刺槐人工林生态系统生物量为34.13—133.08t/hm~2,不同区域之间各组分生物量存在显著性差异。植被层平均碳含量为221.93—454.67 g/kg,总体上表现为乔木层平均碳含量高于灌、草层,枯落物层平均碳含量最低,不同区域乔木、灌木、草本平均碳含量均存在显著性差异。刺槐人工林生态系统碳密度均值为106.86 t/hm~2,其中土壤层碳密度占刺槐人工林生态系统总碳密度的64.09%,是刺槐人工林生态系统碳密度的主要组成部分。植被层碳密度为38.68 t/hm~2,其中乔木层碳密度(33.88 t/hm~2)占植被层碳密度的87.58%,灌木、草本、枯落物所占比例依次为1.98%(0.77 t/hm~2)、2.00%(0.77 t/hm~2)、8.43%(3.26 t/hm~2)。不同区域土壤、生态系统碳密度均存在显著性差异。相关性分析和主成分分析表明,刺槐人工林生态系统碳密度与林龄、降水量呈显著正相关关系,与林分密度、平均气温、海拔和坡度的相关关系不显著,上述林分因子、地形因子和环境因子转化的主成分方差累积贡献率为91.07%,其中林龄和降水量是影响刺槐人工林生态系统碳密度的主要因子,方差贡献率为37.22%。     

黄土高原不同气候区刺槐林恢复年限对水、碳及植物多样性的影响

分析植被恢复对黄土高原人工林生态系统服务的影响,对黄土高原新一轮植被恢复和可持续发展有重要意义。通过文献搜索对已发表的92篇文献中的241组对照试验结果运用整合分析方法,以黄土高原不同气候区内不同恢复年限的刺槐林为代表,选取土壤水分、碳和植物多样性三个指标评价黄土高原刺槐林的生态系统服务变化。结果表明:与对照相比,种植刺槐林后整体上显著提高了土壤碳储量和植物多样性,相对增长率分别为81.30%和32.60%(P<0.05),而土壤储水量显著下降,相对变化率为-35.58%(P<0.05);半湿润地区土壤储水量相对变化率(-22.98%)(P<0.05)和植物多样性相对增长率(149.1%)(P<0.05)显著高于半干旱地区,土壤碳储量相对增长率(57.21%)(P<0.05)低于半干旱地区;刺槐林土壤储水量、碳储量和植物多样性的相对变化率随恢复年限增加依次为-27.57%、-41.80%、35.92%,16.84%、78.58%、156.27%和51.38%、26.37%、27.56%。     

广州红锥-马占相思林物种组成与多样性研究

为了解广州市龙眼洞林场红锥(Castanopsis hystrix)-马占相思(Acacia mangium)阔叶混交林的群落结构,对其物种多样性进行了分析.结果表明,红锥-马占相思阔叶混交林的物种丰富度依次为乔木层>灌木层>草本层>层间植物层.乔木层的Simpson和Shannon-Wiener指数分别为0.945和...     

黄土高原刺槐叶片-土壤生态化学计量参数对降雨量的响应特征

以黄土高原10个典型样点的20年刺槐人工林为研究对象,测定了样地中刺槐叶片碳（C）、氮（N）、磷（P）含量,以及0-10 cm和10-20 cm土层土壤的主要理化指标,探讨了黄土高原刺槐叶片-土壤生态化学计量特征参数随水分梯度的变化规律。研究结果发现：①随着降水量增加,刺槐叶C、N含量呈增加趋势,叶片P含量无一致性规律。② 0-10 cm土层中土壤SOC、TN、TP、铵态氮含量以及C ∶ P和N ∶ P均随降雨量增加呈逐渐增加趋势,而C ∶ N比变化不明显。③刺槐叶片C、N、P及其化学计量比与土壤化学计量比相关性较弱,但叶片N、P与土壤铵态氮、速效磷等速效养分含量呈显著正相关（P<0.05）。本研究叶片N ∶ P>16,说明黄土高原刺槐生长主要受P元素限制,且随着降水的增加,土壤P养分的限制加强。     

珍稀濒危植物江南油杉群落乔木层主要树种种间关联性分析

基于2×2列联表,采用方差比率法(R_v)、x²检验、Pearson相关系数检验和Spearman秩相关系数检验等方法,对福建君子峰国家级自然保护区江南油杉(Keteleeria fortunei var. cyclolepis)群落乔木层主要树种进行种间关联性分析,并通过主成分分析(PCA)划分生态种组。结果表明,21种主要树种总体表现为不显著负关联,且江南油杉与其他物种多呈不显著相关。210个种对中,正相关的有110对,负相关的有81对,不相关的有19对,正负比1.36。绝大多数种对间呈不显著相关,显著率仅为0.48%,物种间相互独立。Pearson相关系数和Spearman秩相关系数检验结果具有一致性,负相关种对数大于正相关种对数,绝大多数种对不显著,显著率分别为6.19%和7.14%。与x²检验相比,Pearson相关系数和Spearman秩相关系数检验方式更加灵敏。运用PCA二维排序可将主要树种划分为4个生态种组,同一生态种组内物种的生态习性和资源需求相似,多呈正相关,种对间相互依赖共存;不同生态种组间物种多呈负...     

Influence of Forest Types and Effects of Forestry Activities on Species Richness and Composition of Chrysomelidae in the Central Mountainous Region of Japan

Species richness and composition of the Chrysomelidae (Coleoptera) were studied in larch (Larix kaempheri [Lamb.] Carrière) plantations, secondary forests, and primary forests. In addition, the effects of forest management practices, such as thinning and long rotation, were examined in the larch plantation. The species richness of Chrysomelidae was higher in the larch plantation than in the secondary forest or in the primary forest. Among the larch plantations, the species richness in old-aged plantations was higher than that in middle-aged plantations. The composition of the beetle assemblages in the larch plantation differed from that in the secondary forest or in the primary forest. Exosoma akkoae (Chujo), Batophila acutangula Heikertinger, and Calomicrus nobyi Chujo were caught with a bias toward the larch plantation. Longitarsus succineus (Foudras) and Sphaeroderma tarsatum Baly were caught more in the secondary forest and the primary forest, respectively. More B. acutangula and S. tarsatum were caught in stands where their host plants occurred at higher rates. Species richness of understory plants was an important factor for chrysomlid species richness, and frequency of host occurrence affected the number of individuals of leaf beetles examined. It seems that forest types and forest management practices affect host plants as well as Chrysomelidae, and that these effects on the host plants also influence chrysomelid assemblages.     

Flavonoid glycosides of the black locust tree, Robinia pseudoacacia (Leguminosae)

Four flavone glycosides isolated from extracts of the leaves of Robinia pseudoacacia (Leguminosae) were characterised by spectroscopic and chemical methods as the 7-O-β-d-glucuronopyranosyl-(1 → 2)[α-l-rhamnopyranosyl-(1 → 6)]-β-d-glucopyranosides of acacetin (5,7-dihydroxy-4′-methoxyflavone), apigenin (5,7,4′-trihydroxyflavone), diosmetin (5,7,3′-trihydroxy-4′-methoxyflavone) and luteolin (5,7,3′,4′-tetrahydroxyflavone). Assignment of glycosidic ¹H and ¹³C resonances in their NMR spectra was facilitated by ²J_HC correlations detected using the H2BC (heteronuclear two-bond correlation) pulse sequence. Spectroscopic analysis of two known triglycosides, acacetin 7-O-β-d-glucopyranosyl-(1 → 2)[α-l-rhamnopyranosyl-(1 → 6)]-β-d-glucopyranoside (previously unrecorded from this species) and acacetin 7-O-β-d-xylopyranosyl-(1 → 2)[α-l-rhamnopyranosyl-(1 → 6)]-β-d-glucopyranoside (‘acacetin trioside’), enabled inconsistencies in the literature relating to these structures to be resolved. Comparison of the flavonoid chemistry of leaves and flowers of R. pseudoacacia using LC-UV and LC-MS showed that flavone 7-O-glycosides, particularly of acacetin, predominated in the former, whereas the latter comprised mainly flavonol 3,7-di-O-glycosides, including several examples new to this species. Tissue dependent differences in flavonoid chemistry were also evident from the glycosylation patterns of the compounds.