A standardized bioassay for evaluation of potential germination stimulants for seeds of parasitic weeds

A standard bioassay for the evaluation of the biological activity of potential germination stimulants for seeds of parasitic weeds has been designed by carefully reconsidering all steps of a literature procedure. Addition of a surfactant to the solution for sterilization of the seeds gave a considerable reduction of contamination during the conditioning and stimulation process. Furthermore, it was shown that the water/seed ratio during conditioning affects the germination results. For the seed conditioning and stimulation process, the sandwich technique was introduced, whereby the seeds were placed between two layers of glass fiber filter paper discs. The standardized bioassay has been used for the evaluation of the stimulatory activity of numerous (new) synthetic analogues of strigol (the natural germination stimulant). It is essential to include a reference compound—GR24, an analogue of strigol, is recommended—in every test series, because it was observed that, although a standard bioassay was used, germination percentages obtained with GR24 solutions vary from test to test. For seeds of Striga hermonthica, seasonal effects were found in the germination percentages obtained with GR24 as stimulant. Test results with strigol analogues having modifications in ring D reveal that structural variations in this part of the molecule have dramatic effects on the biological activity of strigol analogues.     

A concise synthesis of optically active solanacol,the germination stimulant for seeds of root parasitic weeds

Solanacol, isolated from tobacco (Nicotiana tabacum L.), is a germination stimulant for seeds of root parasitic weeds. A concise synthesis of optically active solanacol has been achieved by employing enzymatic resolution as a key step.     

The strigolactone germination stimulants of the plant-parasitic Striga and Orobanche spp. are derived from the carotenoid pathway

The seeds of parasitic plants of the genera Striga and Orobanche will only germinate after induction by a chemical signal exuded from the roots of their host. Up to now, several of these germination stimulants have been isolated and identified in the root exudates of a series of host plants of both Orobanche and Striga spp. In most cases, the compounds were shown to be isoprenoid and belong to one chemical class, collectively called the strigolactones, and suggested by many authors to be sesquiterpene lactones. However, this classification was never proven; hence, the biosynthetic pathways of the germination stimulants are unknown. We have used carotenoid mutants of maize (Zea mays) and inhibitors of isoprenoid pathways on maize, cowpea (Vigna unguiculata), and sorghum (Sorghum bicolor) and assessed the effects on the root exudate-induced germination of Striga hermonthica and Orobanche crenata. Here, we show that for these three host and two parasitic plant species, the strigolactone germination stimulants are derived from the carotenoid pathway. Furthermore, we hypothesize how the germination stimulants are formed. We also discuss this finding as an explanation for some phenomena that have been observed for the host-parasitic plant interaction, such as the effect of mycorrhiza on S. hermonthica infestation.     

Strategies for the synthesis of canonical,non-canonical and analogues of strigolactones,and evaluation of their parasitic weed germination activity

Phytochemistry Reviews - Strigolactones (SLs) are natural products with promising applications as agrochemicals to prevent infestation with parasitic weeds due to their ability to trigger seed...     

Confirmation and quantification of strigolactones, germination stimulants for root parasitic plants Striga and Orobanche, produced by cotton

The germination stimulants for root parasitic plants Striga and Orobanche produced by cotton (Gossypium hirsutum L.) were examined in detail. Seeds of cotton were germinated and grown on glass wool wetted with sterile distilled water in sterile filter units. The root exudate was collected daily and extracted with ethyl acetate. Each of these ethyl acetate extracts was analyzed directly by high-performance liquid chromatography linked with tandem mass spectrometry (LC/MS/MS). The results demonstrate that cotton roots exuded strigol and strigyl acetate, but no other known strigolactones such as orobanchol and alectrol. The production of strigol was detected even in the root exudate collected during the first 24 h of incubation and reached a maximum 5-7 days later. The average exudation of strigol and strigyl acetate during the incubation period was ca. 15 and 2 pg/plant/day, respectively, indicating that strigol mainly contributed to germination stimulation by the cotton root exudate.     

Concise synthesis of heliolactone,a non-canonical strigolactone isolated from sunflower

ABSTRACT

Heliolactone is one of the earliest identified non-canonical strigolactones. Its concise synthesis was achieved by employing Knoevenagel-type condensation and semi-reduction of a malonate intermediate as the key steps. This synthesis was performed in a non-stereoselective manner, and thus a racemic and diastereomeric mixture of heliolactone was obtained. The developed synthetic route is fairly concise and straightforward.     

Design, synthesis, and cytostatic activity of novel cyclic curcumin analogues

A series of novel cyclic analogues of curcumin were synthesized and analyzed for in vitro cytostatic activity. Condensation of 2-acetylcycloalkanones with a variety of aromatic aldehydes resulted in the formation of 2-arylidene-6-(3-arylacryoyl)-cycloalkanone derivatives. A number of these analogues were found to have significant anticancer activity against representative murine and human cancer cell lines during in vitro bioassays. This corroborated with in vitro cytostatic activity against a panel of 60 cell lines studied at the National Cancer Institute (USA).     

Stimulation of gibberellin-induced germination in lettuce seeds by cyclic 3',5'-adenosine monophosphate

Cyclic 3',5'-adenosine monophosphate and sodium dibutyryl cyclic3',5'-adenosine monophosphate had no effect on dark-germinationof light-sensitive lettuce {Lacluca sativa L., var. Grand Rapids)seeds when given alone. Cyclic 3',5'-adenosine monophosphate,however, synergistically enhanced the 3 µ 10^–5 Mgibberellic acid-induced germination, but showed no such effecton kinetin-stimulated germination. 5'-AMP, in contrast to cyclic3',5'-adenosine monophosphate, had no effect on gibberellicacidinduced germination. (Received August 8, 1971; )     

PCR-based sensitive detection of Ralstonia solanacearum from soil,eggplant, seeds and weeds

Ralstonia solanacearum is an economically important, bacterial plant pathogen which affects a wide range of crop plants. R. solanacearum survives in the soil for many years and weeds serve as symptomless carrier. One of the important aspects in controlling R. solanacearum is its early detection. In this study, detection threshold of R. solanacearum in the soil was standardised using polymerase chain reaction (PCR) method. The minimum threshold limit ranged between 6.8 × 10 and 3.6 × 10² CFU g^?1 of soil. Using this standardised protocol R. solanacearum was detected from the rhizosphere soil of eggplants showing varying degrees of wilt. PCR method was quite sensitive to detect R. solanacearum from the xylem fluid of eggplant. Presence of R. solanacearum in the soil infected with capsicum wilt was also demonstrated successfully and the minimum detection limit was 4 × 10² CFU g^?1 of soil. The bacterium was not detected from the eggplant seeds collected during 2006 and 2007 seasons. However, the bacterium was detected from the weed (Alternanthera sessilis) grown in the eggplant field indicating the possibility of weeds serving as symptomless carrier. Using our method, it is possible to detect R. solanacearum from soil, plant and weeds grown in the field at an early stage so that proper management strategies could be taken to prevent the infection and further spread of the pathogen.     

Fabacyl acetate, a germination stimulant for root parasitic plants from Pisum sativum

A germination stimulant, fabacyl acetate, was purified from root exudates of pea (Pisum sativum L.) and its structure was determined as ent-2′-epi-4a,8a-epoxyorobanchyl acetate [(3aR,4R,4aR,8bS,E)-4a,8a-epoxy-8,8-dimethyl-3-(((R)-4-methyl-5-oxo-2,5-dihydrofuran-2-yloxy)methylene)-2-oxo-3,3a,4,5,6,7,8,8b-decahydro-2H-indeno[1,2-b]furan-4-yl acetate], by 1D and 2D NMR spectroscopic, ESI- and EI-MS spectrometric, X-ray crystallographic analyses, and by comparing the ¹H NMR spectroscopic data and relative retention times (RR_t) in LC-MS and GC-MS with those of synthetic standards prepared from (+)-orobanchol and (+)-2′-epiorobanchol. The ¹H NMR spectroscopic data and RR_t of fabacyl acetate were identical with those of an isomer prepared from (+)-2′-epiorobanchol except for the opposite sign in CD spectra. This is the first natural ent-strigolactone containing an epoxide group. Fabacyl acetate was previously detected in root exudates of other Fabaceae plants including faba bean (Vicia faba L.) and alfalfa (Medicago sativa L.).     

Exogenous amino acids inhibit seed germination and tubercle formation by Orobanche ramosa (Broomrape): Potential application for management of parasitic weeds

Parasitic plants are among the most problematic weeds that are responsible for major losses to many crops. Early growth stages, such as seed germination stimulated by host root exudates and tubercle development, are key phases for the development of these parasites. Inhibition of these early phases by naturally occurring compounds could be a general strategic option for management of parasitic plants. In this study, we report that certain natural amino acids cause severe physiological disorders of germinating broomrape seeds. In particular, methionine was able to inhibit almost totally the germination of seeds of Orobanche ramosa when applied at a concentration of 2 mM. Applied to tomato roots, methionine strongly reduced the number of developing tubercles of the parasite. These findings suggest that: (1) appropriate amino acids applied exogenously to a root zone might result in control of parasitic plants such as Orobanche sp.; (2) amino acid excreting microbes introduced into the crop rhizosphere might control root parasitism; and (3) amino acid producing broomrape pathogens might be selected in order to enhance their virulence.     

QTL mapping for germination of seeds obtained from previous wheat generation under drought

The QTLs controlling germination and early seedling growth were mapped using seeds acquired from mapping population and parental lines of Chinese Spring and SQ1 grown under water-limited conditions, severe drought (SDr) and well-watered plants (C). Germination ability was determined by performing a standard germination test based on the quantification of the germination percentage (GP24) of seeds incubated for 24 h at 25°C in the dark. Early seedling growth was evaluated on the basis of the length of the root and leaf at the 6th day of the experiment. QTLs were identified by composite interval mapping method using Windows QTLCartographer 2.5 software. For the traits studied, a total of thirty eight additive QTLs were identified. Seventeen QTLs were mapped in C on chromosomes: 1A, 2A, 7A, 1B, 2B, 3B, 4B, 5B, 6B, 7B, 2D, 3D, 4D and 6D, while twenty one QTLs were identified in SDr on chromosomes: 1A, 2A, 5A, 2B, 3B, 4B, 5B, 6B, 7B, 3D, 5D and 6D. Most of the QTLs for GP and early leaf growth parameters were clustered on chromosome 4B (associated with the Rht-B1 marker) both in C and SDr plants. The results indicate the complex and polygenic nature of germination.     

High-resolution structure of a potent, cyclic proteinase inhibitor from sunflower seeds.

Proteinaceous serine proteinase inhibitors are widespread throughout the plant kingdom where they play an important role in protection against pests and pathogens. Here, we describe the isolation and characterisation of a novel 14 amino acid residue cyclic peptide from sunflower seeds, which is a potent inhibitor of trypsin (Ki=100 pM). The crystal structure of this peptide in complex with bovine beta-trypsin shows both sequence and conformational similarity with the trypsin-reactive loop of the Bowman-Birk family of serine proteinase inhibitors. This inhibitor, however, is unique in being monofunctional, cyclic and far shorter (14 amino acid residues) than inhibitors belonging to this family (typically 60-70 amino acid residues). The high potency of this peptide is likely to arise from the considerable structural rigidity achieved through its cyclic nature which is further stabilised by a single internal disulphide bond. This study helps delineate the minimal unit required for effective peptide inhibitors of serine proteinases, and will assist in the further design of inhibitors to this widespread class of enzymes.     

Isolation of lignans as seed germination and plant growth inhibitors from Mediterranean plants and chemical synthesis of some analogues

Lignans and lignins are among the main metabolic products of phenylpropanoid metabolism in vascular plants. They are compounds representing the building blocks of plant cell walls. Moreover they have a broad range of biological activities such as antitumoral, antimitotic, antiviral and cytotoxic and are thought to be involved in the plant defense against pathogens and pests. In this paper we report a survey of the past and current literature about lignans and neolignans and their germination inhibitory activity on cultivated and wild species from plants of the Mediterranean area. Some examples of synthetic methodologies of these molecules have also been reported.     

温度对四川小金县岷江柏种子发芽的影响

通过对采自四川小金县的岷江柏(Cupressus chenggiana S. Y. Hu)种子发芽特征的研究,分析不同温度对种子发芽的影响.结果表明,采自四川小金县的岷江柏种子发芽周期为20 d,其中0 ～5 d为萌动期,5 ～15 d为高峰期;发芽温度为5℃～30℃,适宜发芽温度为10℃～25℃,种子最适发芽温度在2003年1、4、7和10月分别为25℃、15℃、20℃和20℃.岷江柏种子的最适发芽温度和贮藏过程中的发芽特征与环境温度有着密切的关系.     

Isolation and identification of alectrol as (+)-orobanchyl acetate, a germination stimulant for root parasitic plants

Alectrol, a germination stimulant for root parasitic plants, was purified from root exudates of red clover (Trifolium pratense L.) and identified as a strigolactone, (+)-orobanchyl acetate [(3aS,4S,8bS,E)-8,8-dimethyl-3-(((R)-4-methyl-5-oxo-2,5-dihydrofuran-2-yloxy)methylene)-2-oxo-3,3a,4,5,6,7,8,8b-octahydro-2H-indeno[1,2-b]furan-4-yl acetate], by 1D and 2D NMR spectroscopy and ESI- and EI-MS spectrometry. Orobanchyl acetate afforded an [M-42](+) ion in EI-MS and thus had been recognized as an isomer of strigol. Orobanchyl acetate was detected in root exudates of soybean (Glycine max L.) and cowpea (Vigina unguiculata L.) along with orobanchol.     

Chemical synthesis and kinetic study of the smallest naturally occurring trypsin inhibitor SFTI-1 isolated from sunflower seeds and its analogues

The smallest known naturally occurring trypsin inhibitor SFTI-1 (14 amino acid residues head-to-tail cyclic peptide containing one disulfide bridge) and its two analogues with one cycle each were synthesized by the solid phase method. Their trypsin inhibitory activity was determined as association equilibrium constants (K(a)). Additionally, hydrolysis rates with bovine beta-trypsin were measured. Among all three peptides, the wild SFTI-1 and the analogue with the disulfide bridge only had, within the experimental error, the same activity (the K(a) values 1.1 x 10(10) and 9.9 x 10(9) M(-1), respectively). Both peptides displayed unchanged inhibitory activity up to 6 h. The trypsin inhibitory activity of the analogue with the head-to-tail cycle only was 2.4-fold lower. It was also remarkably faster hydrolyzed (k = 1.1 x 10(-4) mol(peptide) x mol(enzyme)(-1) x s(-1)) upon the incubation with the enzyme than the other two peptides. This indicates that the head-to-tail cyclization is significantly less important than the disulfide bridge for maintaining trypsin inhibitory activity.     

A three-residue cyclic scaffold of non-RGD containing peptide analogues as platelet aggregation inhibitors: design, synthesis, and structure--function relationships

Antagonists of fibrinogen at the GPIIb/IIIa receptor, which is the most abundant membrane protein on the platelet surface, are under active investigation as potential antithrombotics. The critical interaction between GPIIb/IIIa and fibrinogen can be inhibited by either linear or cyclic RGDS-containing peptides, which have been proved as lead compounds in the design of platelet aggregation inhibitors. In this study we present the design and construction of a new class of cyclic (S,S) non-RGD containing peptide sequences, using two Cys as a structural scaffold for the development of antiaggregatory agents. The (S,S)-CDC- sequence was incorporated as a conformational constraint, in molecules bearing at least one positive charge with the general formula (S,S)XCDCZ, where X = Ac-Arg, Pro-Arg, Pro-Ser-Lys, and Pro-Ser-Arg, and Z = -NH(2) and Arg-NH(2). Investigation of the structure-function relationships was performed on the basis of (a) the local conformation induced by the (S,S)-CDC motif, (b) the distance of the positively (R-C(zeta) or K-N(zeta)) and negatively (D-C(gamma)) charged centers, (c) the presence of a second positive or negative charge on the molecule, and (d) the orientation of the basic and acidic side chains defined by the pseudo dihedral angle (Pdo), which is formed by the R-C(zeta), R-C(alpha), D-C(alpha), and D-C(gamma) atoms in the case of (S,S)-RCDC and by the K-N(zeta), K-C(alpha), D-C(alpha), and D-C(gamma) atoms in the case of (S,S)-KCDC.