Fatty acid composition of seed oil triglycerides inCoincya (Brassicaceae)

Oil and triglyceride contents and fatty acid composition were determined for seeds in nine taxa belonging to the genusCoincya (Brassicaceae) on the Iberian Peninsula (Spain and Portugal). The oil content ranges from 11.1 to 24.6%, triglycerides from 68.7 to 88.5%. The major fatty acids were erucic (24.6–30.5%), linolenic (17.7–27.7%), linoleic (13.9–24.6%) and oleic acid (12.3–21.8%).     

A role for isothiocyanates in plant resistance against the specialist herbivore Pieris rapae

We experimentally reanalyzed the classic interaction between Pieris rapae, a specialist lepidopteran herbivore, and isothiocyanates (mustard oils) that are characteristic phytochemicals of the Brassicaceae. Previous investigations have suggested that P. rapae is unaffected by isothiocyanates. Using whole plants, root extracts, and a microencapsulated formulation of allyl isothiocyanate, we now show that isothiocyanates reduce herbivore survival and growth, and increase development time, each in a dose-dependent manner. Neither the substrate allyl glucosinolate, nor myrosinase, the enzyme that results in the breakdown of glucosinolates, negatively affected P. rapae. Thus, we present strong evidence for a role for isothiocyanates in plant resistance against the specialist herbivore P. rapae.     

Orientation and Feeding Responses of the Pollen Beetle, Meligethes aeneus, to Candytuft, Iberis amara

The pollen beetle, Meligethes aeneus, which is an important pest of oilseed rape, Brassica napus, and turnip rape, B. rapa var. campestris, does not oviposit in all species of the Brassicaceae. The relationship between M. aeneus and candytuft, Iberis amara (Brassicacae), was investigated as part of chemical ecological studies into the development of control methods employing non-host-derived repellents. In choice and nonchoice feeding tests, M. aeneus completely rejected I. amara. However, in a field experiment using traps baited with flowering racemes of I. amara and B. napus, M. aeneus was attracted to both species. Gas chromatographic (GC) and GC-electroantennogram (GC-EAG) analyses indicated that the profiles of the floral volatiles of the two species are different. At least 12 compounds among the I. amara floral volatiles were detected by the M. aeneus antenna, and, of these, hexanoic acid, (E)-4,8-dimethyl-1,3,7-nonatriene and alpha-cedrene were not found among B. napus flower volatiles. Since M. aeneus is stimulated by floral volatiles to approach I. amara, but rejects it near, or at, the plant surface, I. amara does not produce repellents that could be used to manipulate M. aeneus. However, it may contain feeding deterrent(s) that could be used in "push-pull" control techniques or in the development of resistant brassicaceous crops.     

Sequestration of Host Plant Glucosinolates in the Defensive Hemolymph of the Sawfly Athalia rosae

Interactions between insects and glucosinolate-containing plant species have been investigated for a long time. Although the glucosinolate–myrosinase system is believed to act as a defense mechanism against generalist herbivores and fungi, several specialist insects use these secondary metabolites for host plant finding and acceptance and can handle them physiologically. However, sequestration of glucosinolates in specialist herbivores has been less well studied. Larvae of the turnip sawfly Athalia rosae feed on several glucosinolate-containing plant species. When larvae are disturbed by antagonists, they release one or more small droplets of hemolymph from their integument. This reflex bleeding is used as a defense mechanism. Specific glucosinolate analysis, by conversion to desulfoglucosinolates and analysis of these by high-performance liquid chromatography coupled to diode array UV spectroscopy and mass spectrometry, revealed that larvae incorporate and concentrate the plant's characteristic glucosinolates from their hosts. Extracts of larvae that were reared on Sinapis alba contained sinalbin, even when the larvae were first starved for 22 hr and, thus, had empty guts. Hemolymph was analyzed from larvae that were reared on either S. alba, Brassica nigra, or Barbarea stricta. Leaves were analyzed from the same plants the larvae had fed on. Sinalbin (from S. alba), sinigrin (B. nigra), or glucobarbarin and glucobrassicin (B. stricta) were present in leaves in concentrations less than 1 mol/g fresh weight, while the same glucosinolates could be detected in the larvae's hemolymph in concentrations between 10 and 31 mol/g fresh weight, except that glucobrassicin was present only as a trace. In larval feces, only trace amounts of glucosinolates (sinalbin and sinigrin) could be detected. The glucosinolates were likewise found in freshly emerged adults, showing that the sequestered phytochemicals were transferred through the pupal stage.     

Histochemical characterisation of unextractable seed coat pigments and quantification of extractable lignin in the Brassicaceae

The differences determined by in situ histochemical staining and thioglycolic lignin analyses provided a new view of seed coat composition in a range of yellow‐ and brown‐seeded germplasm of the Brassicaceae. Unextractable seed coat pigments were composed of proanthocyanidin (condensed tannin) polymers. Anthocyanins were absent in all seed coat tissues. Proanthocyanidin was deposited over the entire seed coat in dark‐seeded germplasm and in patches in some yellow‐seeded germplasm. The seed coat and hilum of Crambe abyssinica cv Prophet and some yellow‐seeded accessions contained an unidentified, unreactive brown pigment. Several new pigmentation patterns, occurring as spots and patches and surrounding the hilum, became apparent from this analysis. Thioglycolic lignin was significantly lower in yellow‐seeded samples compared with the dark‐seeded accessions, and the majority of the lignin occurred in the seed coat. These discoveries provide new and useful data for plant breeders and indicate the need to use more detailed analytical methods in breeding programmes that have the goal of improving seed meal quality by reducing fibre and phenolics. Copyright © 2004 Society of Chemical Industry     

Phytohormonal and metabolism analysis of <i>Brassica rapa</i> L. ssp. <i>pekinensis</i> with different resistance during <i>Plasmodiophora brassicae</i> infection

Clubroot of Chinese cabbage (Brassica rapa L. ssp. pekinensis), caused by the obligate parasite Plasmodiophora brassicae, accounts for serious yield losses. The aim of our study was to explore the phytohormone levels and metabolome changes in the roots of resistant and susceptible B. rapa genotypes at a late stage of infection, i.e., 28 days post-infection. Both genotypes showed decreased auxin levels after P. brassicae infection except for indole-3-acetic acid. Overall, the susceptible genotype had higher auxin and cytokinin levels after infection, with the exception of trans-zeatin and 3- indolebutyric acid as compared to the resistant genotype. Jasmonic acid levels declined after infection regardless of the genotype. Resistance against clubroot was evident with the increased levels of salicylic acid in the resistant genotype. The susceptible genotype had a higher number of differentially accumulated metabolites (DAMs) (262) than the resistant genotype (238) after infection. Interestingly, 132 DAMs were commonly detected in both genotypes when infected with the pathogen, belonging to metabolite classes such as phenolic acids, amino acids, and derivatives, glucosinolates, organic acids, flavonoids, nucleotides and derivatives, and fatty acids. The differential metabolite analysis revealed that metabolites related to amino acid biosynthesis, fatty acid biosynthesis and elongation, glutathione metabolism, and glucosinolate metabolism were highly accumulated in the resistant genotype, suggesting their essential roles in resistance against P. brassicae infection.     

Identification of desulphoglucosinolates in Brassicaceae by LC/MS/MS: comparison of ESI and atmospheric pressure chemical ionisation-MS

In order to develop a sensitive method for the detection of desulphoglucosinolates by HPLC-MS, the two most common interfaces for HPLC-MS, atmospheric pressure chemical ionisation (APCI) and ESI, were compared. While working with the APCI-interface the evaporation temperature and corona amperage were optimised. In doing so 300 degrees C and 6 muA proved to be most suitable for aliphatic and indole desulphoglucosinolates. The use of formic acid instead of water in the eluent in HPLC-ESI-MS measurements increased the sensitivity for the indole desulphoglucosinolates in the presence of 1 mM formic acid, while the sensitivity for the aliphatic desulphoglucosinolate desulphoglucoraphanin was substantially increased by the presence of 5 mM formic acid. Using an Agilent ion trap, two optimisation procedures for the MS parameters, smart and expert mode, were available. In smart mode the software optimises several parameters automatically, which is much more time efficient than expert mode, in which the optimisation is done manually. It turned out that ESI-MS is most sensitive in smart mode, while for APCI-MS a higher sensitivity could be gained using the expert mode. Comparing both interfaces, APCI-MS was more sensitive than ESI-MS. However, no additional information, in terms of structure determination, was obtained by APCI-MS.     

Homology-Based Interactions between Small RNAs and Their Targets Control Dominance Hierarchy of Male Determinant Alleles of Self-Incompatibility in Arabidopsis lyrata

Self-incompatibility (SI) is conserved among members of the Brassicaceae plant family. This trait is controlled epigenetically by the dominance hierarchy of the male determinant alleles. We previously demonstrated that a single small RNA (sRNA) gene is sufficient to control the linear dominance hierarchy in Brassica rapa and proposed a model in which a homology-based interaction between sRNAs and target sites controls the complicated dominance hierarchy of male SI determinants. In Arabidopsis halleri, male dominance hierarchy is reported to have arisen from multiple networks of sRNA target gains and losses. Despite these findings, it remains unknown whether the molecular mechanism underlying the dominance hierarchy is conserved among Brassicaceae. Here, we identified sRNAs and their target sites that can explain the linear dominance hierarchy of Arabidopsis lyrata, a species closely related to A. halleri. We tested the model that we established in Brassica to explain the linear dominance hierarchy in A. lyrata. Our results suggest that the dominance hierarchy of A. lyrata is also controlled by a homology-based interaction between sRNAs and their targets.