Extracts and purified substances of Cabralea canjerana inhibit hatching and extracts of Schinus terebinthifolius kill juveniles of meloidogyne incognita

Nematicidal substances have been identified from plants and are potentially useful for the management of plant-parasitic nematodes. Cabralea canjerana, (Meliaceae) and Schinus terebinthifolius (Anacardiaceae) produce bioactive compounds during their secondary metabolism and little is known about the effect of such substances on plant-parasitic nematodes. In the present study, we assessed the effect of aqueous and ethanolic extracts of C. canjerana and S. terebinthifolius at 1% (m:v) and purified substances from C. canjerana (gedunin, ocotillone, cabraleadiol, a mixture of ocotillone + cabraleadiol and a mixture of shoreic acid + eichlerianic acid) on hatching and mortality of Meloidogyne incognita juveniles. Aqueous extracts of C. canjerana fruits and seeds reduced hatching by 70.3 to 95.7%. Aqueous extracts of S. terebinthifolius fruits killed 42.8 to 77.1% of juveniles. The purified substances of C. canjerana inhibited the hatching of M. incognita from 57 to 90% and did not increase the mortality of juveniles. Therefore, C. canjerana extracts and its purified substances reduce M. incognita hatching and aqueous extracts of S. terebinthifolius kill J2 of this nematode.

Rapid differentiation of graft Citrus sinensis with and without Xylella fastidiosa infection by mass spectrometry.

RATIONALE: Xylella fastidiosa causes citrus variegated chlorosis (CVC) in sweet orange trees. A diagnostic method for detecting CVC before the symptoms appear, which would inform citrus producers in advance about when the plant should be removed from the orchard, is essential for reducing pesticide application costs. METHODS: Chemometrics was applied to high-performance liquid chromatography diode array detector (HPLC-DAD) data to evaluate the similarities and differences between the chromatographic profiles. A liquid chromatography/atmospheric pressure chemical ionization mass spectrometry selected reaction monitoring (LC/APCI-MS-SRM) method was developed to identify the major compounds and to determine their amounts in all samples. RESULTS: We evaluated the effect of this bacterium on the variation in the chemical profile in citrus plants. The organs of C. sinensis grafted on C. limonia were analyzed. Chemometrics was applied to the obtained data, and two major groups were differentiated. Flavonoids were observed in one group (leaves) and coumarins in the second (roots), both at higher concentrations in the plants with CVC symptoms than in those without the symptoms and those in the negative control. The rootstocks also interfered in the metabolism of the scion. CONCLUSIONS: The developed LC/APCI-MS-SRM method for detecting CVC before the symptoms appear is simple and accurate. It is inexpensive, and many samples can be screened per hour using 1 mg of leaves. Knowledge of the influence of the rootstock on the chemical profile of the graft is limited. This study demonstrates the effect of the rootstock in synthesizing flavonoids and increasing its content in all parts of the graft.

Quantification and localization of hesperidin and rutin in Citrus sinensis grafted on C. limonia after Xylella fastidiosa infection by HPLC-UV and MALDI imaging mass spectrometry.

A high performance liquid chromatography-ultraviolet (HPLC-UV) method was developed for quantifying hesperidin and rutin levels in leaves and stems of Citrus limonia, with a good linearity over a range of 1.0-80.0 and 1.0-50.0 µg mL(-1) respectively, with r(2)>0.999 for all curves. The limits of detection (LOD) for both flavonoids were 0.6 and 0.5 µg mL(-1), respectively, with quantification (LOQ) being 2.0 and 1.0 µg mL(-1), respectively. The quantification method was applied to Citrus sinensis grafted onto C. limonia with and without CVC (citrus variegated chlorosis) symptoms after Xylella fastidiosa infection. The total content of rutin was low and practically constant in all analyses in comparison with hesperidin, which showed a significant increase in its amount in symptomatic leaves. Scanning electron microscopy studies on leaves with CVC symptoms showed vessel occlusion by biofilm, and a crystallized material was noted. Considering the difficulty in isolating these crystals for analysis, tissue sections were analyzed by matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) to confirm the presence of hesperidin at the site of infection. The images constructed from MS/MS data with a specific diagnostic fragment ion (m/z 483) also showed higher ion intensities for it in infected plants than in healthy ones, mainly in the vessel regions. These data suggest that hesperidin plays a role in the plant-pathogen interaction, probably as a phytoanticipin. This method was also applied to C. sinensis and C. limonia seedlings, and comparison with the graft results showed that the rootstock had an increased hesperidin content ∼3.6 fold greater in the graft stem than in the stem of C. sinensis seedlings. Increase in hesperidin content by rootstock can be related to induced internal defense mechanisms.

Alkyl, aryl, alkylarylquinoline, and related alkaloids.

The Rutaceae continues to be the primary source of new alkyl-, aryl-, and alkylarylquinolin/ones. In the past 17 years, the overall distribution of these alkaloid types within the family has changed little since the chemosystematics reviews by Waterman (270), Mester (40), and da Silva et al. (279). Alkylquinolones dominate the reported isolations with about 51% of the total, with arylquinolones (16%), alkylquinolines (15%), alkylarylquinolines (11%), arylquinolines (3%), alkylarylquinolones (2%), and quinolines (2%) as the significant structural groups contributing to the remainder of this class of alkaloids. The alkyl-, aryl-, and alkylarylquinolin/one alkaloids occur in 50 species belonging to 24 genera and 6 subfamilies. Despite the intensive chemical exploration of many species from other plants in the Rutales family, but not in the family Rutaceae, the first alkaloid alkylquinolone from a simaroubaceous plant (160) was not reported until 1997. Although many additional alkaloids have been reported, some of new structural types (Bo.4), substantial biosynthetic work on plant-derived alkylquinolin/ones has not yet been carried out. The biosynthesis of some of these alkaloids in bacteria was firmly established as being derived from anthranilic acid. Outside of the Rutales, alkyl-, aryl-, and alkylarylquinolin/ones have not been found, except for simple quinoline (A.1; only one) and 2-methylquinoline derivatives in the Zygophyllaceae, and only an atypical quinolone derivative (Ao.1) in the Asteraceae family. A few 3-phenylquinolines (2), 3-(1H-indol-3-yl)quinoline (1), and quinoline-quinazoline (1) alkaloids have been reported from only a single genus in the Zygophyllaceae. Tryptophan-derived quinolines in higher plants are confined to a few 2-carboxylicquinolin/ones (6) and 4-carbaldehydequinolines (5); the former found in the Ephedraceae (5), Boraginaceae (1), Fagaceae (1), Ginkgoaceae (1), Plumbaginaceae (1), Solanaceae (1), and Apiaceae (1), and the latter in the Moraceae (3), Alliaceae (1), and Pontederiacae (1). The number of quinolones derived from glycine and a polyketide is also limited. 5-Alkyl-2-methylquinolin-4(1H)-ones (8) occur in the Euphorbiaceae, and 5-alkyaryl-2-methylquinolin-4(1H)-ones ((3) in the Sterculiaceae. Alkylquinolin/ones are well-known as typical alkaloids of three Proteobacteria and three Actinobacteria; the genus Pseudomonas yielded the majority (46%) of the total number of alkaloids reported (39). 2-Carboxylicquinolin/ones (4) and 4-carbaldehydequinolines (6) are minor constituents in both divisions of bacteria. More interesting are the quinolactacins (7), in which the second nitrogen is derived from L-valine or L-isoleucine, recently reported to occur only in the fungus Penicillium. Many of these diverse alkaloids have served directly as medicines or as lead compounds for the synthesis (258) of derivatives with an improved biological profile. It is apparent from the summary view of the alkyl-, aryl-, and alkylarylquinolin/ones reported in the Rutaceae that they help to confirm the affinity between Rutoideae tribes and provide firm support for placing the Spathelioideae and the Dictyolomatoideae close to the more primitive Zanthoxyleae tribe. On the other hand, the bacteria and fungi are needed for more substantial chemical studies. When more data become available, it is likely that useful systematic correlations will emerge. More detailed studies regarding the biosynthetic pathways of the alkyl-, aryl-, and alkylarylquinolin/ones in the Rutaceae and in bacteria are needed. Such studies would clarify the differences in the pathways based on their derivation from anthranilic acid in bacteria and in rutaceous plants. Finally, this survey indicates that the Rutaceae, and various bacterial and fungal species offer considerable potential for the discovery of new or known alkaloids with significant and possibly valuable biological activities.