Attachment to plant surface waxes by an insect predator

Insects foraging on plant surfaces must attach to the layerof lipophilic materials known as epicuticular waxes (EW) thatcover these surfaces. In this paper, we briefly review the evidencethat variation in EW morphology can influence the ecology ofherbivorous insects directly, by affecting their attachmentto plant surfaces, and indirectly by affecting attachment byactively foraging predatory insects to plant surfaces. We thenpresent new data examining how EW micromorphology and chemicalcomposition of Brassica oleracea influence attachment by thepredatory beetle, Hippodamia convergens (Coccinellidae). Bioassayswith genotypes of B. oleracea differing in wax characteristics,and with EW extracts from these plants applied to glass, showthat wax crystals disrupt attachment. In addition, bioassaysshow that attachment by H. convergens differs among EW extractsprepared to have smooth surfaces without crystals. The differencesin attachment under these conditions are evidently due to thechemical composition of the waxes. Bioassays with two pure waxconstituents show that wax composition can significantly affectattachment by H. convergens. The study opens the way for usinga similar approach to understand attachment by insects to waxyplant surfaces.     

Genetic Diversity inBrassica oleraceaL. (Cruciferae) and Wild Relatives (2n=18) using RAPD Markers

This paper presents the evaluation of genetic diversity in 29populations of wild taxa of theBrassica oleraceaL. group (2n=18)and two cultivars, using RAPDs as molecular markers. In a previouspaper (Lázaro and Aguinagalde,Annals of Botany82: 000–000,1998), 11 isozymes were used for the same purpose. Results obtainedwith the two molecular markers (isozymes and RAPDs) are compared.DNA from ten individuals per population was analysed using sixdifferent primers; the 151 detected bands were polymorphic,11 were common to all species, six to all taxa, only one toevery population; and no bands were shared by every individual.The dendrogram obtained using genetic distances clustersB. oleraceapopulationswithB. bourgeaui, B. alboglabra, B. montanaandB. incana. B.insularis, B. macrocarpa, B. villosaandB. rupestrispopulationsform another cluster. Populations ofB. creticaandB. hilarionisformthe third cluster. Genetic diversity inB. oleraceapopulations,theB. rupestriscomplex andB. creticasubspecies was estimatedusing the AMOVA programme; the latter was the most diverse.Copyright1998 Annals of Botany Company. Brassica oleraceaL., wild relatives, genetic diversity, genetic resources, RAPD markers, AMOVA.     

Chemical composition and ultrastructure of the epicuticular wax in three lines of Brassica napus (L)

The epicuticular wax in three lines of Brassica napus (rape) has been investigated and the detailed chemistry and ultrastructure of the waxes examined. A distinct chemical make-up has been found for all three waxes which is correlated with three distinct crystallite structures. A tentative scheme for classification of Brassica wax mutants is described in which the two newly analysed rape mutants can be placed. Mass spectral analysis of all wax components confirms and extends previous ideas about the chemistry of Brassica waxes.     

Effect of Different Seed Storage Conditions on Germination and Isozyme Activity in Some Brassica Species

Sixteen seed accessions of four Brassica species (B. cretica,B. incana, B. montana and B. oleracea) were examined following5-22 years' storage in the germplasm bank. Germinability andisozyme of seeds stored under long-term (-10 °C and 3% moisturecontent) and short-term (5% °C and 8% moisture content)storage conditions were compared. Long-term storage producedno deterioration and the ability to germinate was satisfactorilymaintained over 8-22 years. Short-term storage conditions maintainedgermination ability up to 10-12 years in all accessions of B.cretica and B. montana. However, seed multiplication might beessential every 10 years for some accessions of B. oleraceastored in this way. In the 8 to 9-year-old accessions of B.cretica, B. incana and B. montana, no significant differenceswere detected between conservation systems for germinabilityand frequency of seeds showing isozyme activity for seven enzymesystems (ACO, IDH, MDH, ME, PGI, PGM and 6-PGD). However, significantdifferences were found for the ADH enzyme system. Moreover,in the 5-22-year-old B. oleracea accessions, significant differenceswere found between storage conditions with respect to isozymeactivity for all enzyme systems studied.Copyright 1995, 1999Academic Press Brassica cretica, Brassica incana, Brassica montana, Brassica oleracea, accessions, isozymes, germination, seed storage     

Distribution of similar self-incompatibility (<Emphasis Type="Italic">S</Emphasis>) haplotypes in different genera,<Emphasis Type="Italic"> Raphanus</Emphasis> and<Emphasis Type="Italic"> Brassica</Emphasis>

The nucleotide sequences of ten SP11 and nine SRK alleles in Raphanus sativus were determined, and deduced amino acid sequences were compared with those of Brassica SP11 and SRK. The amino acid sequence identity of class-I SP11s in R. sativus was about 30% on average, the highest being 52.2%, while that of the S domain of class-I SRK was 77.0% on average and ranged from 70.8% to 83.9%. These values were comparable to those of SP11 and SRK in Brassica oleracea and B. rapa. SP11 of R. sativus S-21 was found to be highly similar to SP11 of B. rapa S-9 (89.5% amino acid identity), and SRK of R. sativus S-21 was similar to SRK of B. rapa S-9 (91.0%). SP11 and SRK of R. sativus S-19 were also similar to SP11 and SRK of B. oleracea S-20, respectively. These similarities of both SP11 and SRK alleles between R. sativus and Brassica suggest that these S haplotype pairs originated from the same ancestral S haplotypes.     

Modification of the Pollen-Stigma Interaction in Brassica oleracea by Water

The presence of a film of distilled water on the stigma surfaceof freshly opened flowers results in complete inhibition ofpollen following both incompatible and compatible pollinationsin self-incompatible (SI) genotypes of Brassica oleracea, SIgenotypes of B. campestris and one self-compatible (SC) genotypeof B. campestris. The application of water to the stigmas afterpollination also resulted in a marked reduction in pollen germinationand tube penetration. An increase in the time intervals betweenthe application of pollen onto the stigma and the water treatmentprogressively reduced this inhibition. Pollen germination wasalso completely inhibited when stigmas from freshly-opened flowersof SI B. campestris and B. oleracea genotypes were washed inwater, dried and pollinated with pollen grains of either compatibility.The ability of stigmas to induce pollen germination and tubegrowth was restored over a period, the length of which was dependenton the incompatibility (S) genotype. Stigmas of B. napus (SC)and SC mutants of SI B. campestris were found to be affectedby washing, but stigmas of a SC variety of B. campestris andthe immature stigmas from buds of B. oleracea were found tobe considerably less affected. Microscopic examination of pollenplaced on washed stigmas reveals that grains, irrespective oftheir compatibility, fail to hydrate normally. When inducedto hydrate by raising atmospheric humidity, pollen grains onwashed stigmas did germinate, but most of the tubes failed topenetrate the papillar wall and very few entered the style.It is proposed that the water treatment mobilises componentsof the pellicle which reorganize to block the activity of molecules,present in both SC and SI individuals, responsible for establishingfull contact between the pellicle and pollen grain coating. Brassica, pellicle, pollen, recognition, self-incompatibility     

Comparative Studies of the Seed Proteins of Brassica campestris, Brassica oleracea, and Brassica nigra

The albumin and globulin fractions of the seeds of Brassicacampestris, B. oleracea, and B. nigra have been investigatedusing serological methods and separation by acrylamide gel electrophoresisin order to determine the usefulness of these techniques fortaxonomic studies of the genus Brassica. The results of theprotein analysis given in this paper agree with establishedtaxonomy and suggest that both methods may prove useful in varioustaxonomic problems in the genus Brassica.     

Differential abundance of simple repetitive sequences in species ofBrassica and relatedBrassicaceae

SixBrassica species, known as the triangle of U, and four species from related genera were characterized by DNA fingerprinting with simple repetitive oligonucleotide probes. Our results show that CT-, TCC-, and GTG-repeat motifs are equally abundant in the genomes of the sixBrassica species. In contrast, GATA-, GGAT-, and GACA-multimers are unevenly distributed among different species. As judged from the number and strength of hybridization signals, the highest copy number of all three motifs occurs inBrassica nigra, while the lowest is observed inB. oleracea. The abundance of GATA-and GACA-repeats varies in a coordinate way. The amphidiploid genomes ofB. juncea, B. carinata, andB. napus each harbour intermediate amounts of (GATA)₄ and (GACA)₄-detected repeats as compared to their diploid progenitors, thus supporting the concept of the U triangle. GATA-, GACA-, and GGAT-repeats were also abundant inEruca sativa andSinapis arvensis, but not inRaphanus sativus andSinapis alba. These results support the idea thatBrassica nigra is more closely related toSinapis arvensis than to otherBrassica species such asB. rapa andB. oleracea.     

Genetic Diversity inBrassica oleraceaL. (Cruciferae) and Wild Relatives (2n=18) using Isozymes

Genetic diversity in 36 populations of wild taxa of theBrassicaoleraceaL. group (2n=18) and two cultivated forms was studiedusing isozyme variation at 11 loci for five enzyme systems (IDH,6-PGD, PGM, PGI, MDH). Mean values for the percentage of polymorphicloci and expected heterozygosity were 54% and 0.224, respectively.Statistically significant differences among allele frequencieswere found with the 6-PGD isozyme system. Intrapopulationalgenetic diversity was 67% while interpopulational genetic diversitywas only 33%. The dendrogram obtained, using genetic distancesamong taxa, showed three different groups. With the exceptionofB. incana,they agree to the already accepted relationshipsamong the 14 taxa studied: the West Mediterranean group, withB.oleracea, B. alboglabra, B. bourgeauiandB. incana; another groupof species growing in the central Mediterranean area, whichincludesB. villosa, B. villosasubsp.drepanensis, B. rupestris,B. macrocarpa(the four taxa together withB. incanaare consideredtheB. rupestrisgroup) andB. montana; and finally the Aegeangroup, which includes the three subspecies ofB. cretica.Clearlyseparated wereB. insularisandB. hilarionis, showing the maximumgenetic distance. Separate dendrograms were also obtained forB.oleracea, B. montana, B. creticaandB. rupestrisgroup, and geneticdiversity parameters were estimated. Genetic distances amongB.oleraceapopulations are in the same range as populations oftheB. creticasubspecies. Highest genetic distances were foundamong populations of theB. rupestrisgroup.Copyright 1998 Annalsof Botany Company. Brassica oleraceaL., wild relatives, genetic diversity, genetic resources, isozymes.     

Survival and behavior of Plutella xylostella larvae on cabbages with leaf waxes altered by treatment with S-ethyl dipropylthiocarbamate

Cabbages (Brassica oleracea L.) treated with S-ethyl dipropylthiocarbamate (EPTC) herbicide had reduced amounts of leaf surface waxes (40.6% of controls) and reduced densities of leaf surface wax crystallites (20.8% of controls). Leaf waxes of EPTC-treated plants chemically and morphologically resembled leaf waxes of genetically glossy cabbages resistant to the diamondback moth Plutella xylostella (L.) (Lepidoptera: Plutellidae). Survival of larvae was significantly reduced on EPTC-treated cabbage plants in three out of four experiments (62.0–15.3% of survival on controls). P. xylostella neonates also moved more rapidly on EPTC-treated plants than on untreated controls (1.84±0.16 cm/min on controls vs. 3.94±0.24 cm/min on treated plants; P=0.0001). These results support the hypotheses that reduction in leaf waxes is the basis of resistance to P. xylostella in genetically glossy plants and that reduced acceptance by larvae is associated with this resistance. Modification of leaf surface waxes with EPTC or similar compounds may have potential as an economic control for P. xylostella in Brassica crops.     

Metabolic effects of low cortisol during exercise in humans

This studyexamined the physiological effect of reduced plasma cortisol (C) duringprolonged exercise in humans. The effects of normal C (NC) werecompared with metyrapone-induced low C (LC) on plasma substrateavailability and the respiratory exchange ratio during 2 h of exerciseat ~60% peak O₂ consumption innine subjects. The C responses were compared with preexercise (Pre) levels and with a rest day (Con). At rest, C was attenuated by ~70%for LC compared with NC. At rest, plasma glucose, lactate, glycerol,-hydroxybutyrate, alanine, branched-chain amino acids, insulin,glucagon, growth hormone, epinephrine, and norepinephrine were similarunder LC and NC (P > 0.05). Duringexercise under NC, plasma C increased compared with Pre, whereas itremained unchanged during LC. During NC, plasma C was elevated at 90 min (compared with Con) and at 120 min (compared with Con and Pre). During exercise, plasma glucose decreased to the same extent and lactate was similar under both conditions, whereas plasma glycerol, -hydroxybutyrate, alanine, and branched-chain amino acids were higher (P < 0.01) under NC. Plasmainsulin declined (P = 0.01) to agreater extent under LC, whereas growth hormone, epinephrine, andnorepinephrine tended to be higher (0.05  P  0.10). Plasma glucagon increasedunder both conditions (P < 0.01).The respiratory exchange ratio did not differ between conditions. Weconclude that, during exercise, 1) Caccelerates lipolysis, ketogenesis, and proteolysis;2) under LC, glucoregulatory hormoneadjustments maintain glucose homeostasis; and3) LC does not alter whole body substrate utilization or the ability to complete 2 h of moderate exercise.

     

In vitro Shoot Regeneration from Seedling Root Segments of Brassica oleracea and Brassica napus Cultivars

Root segments taken from aseptically-grown seedlings of Brassicaoleracea var. italica cv. Green Comet were used in an investigationof factors affecting in vitro regeneration. Shoot regenerationwas found to increase with seedling age and to be highest inroot segments adjacent to the hypocotyl and lowest in segmentsadjacent to the root tip. In a comparison of a range of mediaand agar concentrations shoot formation was favoured by complexmedia containing reduced nitrogen and was higher on gelled mediathan in liquid medium. The effects of various cytokinins andauxins were investigated; KN was the best cytokinin and IAAand Picloram the best auxins for shoot induction. Root segmentsfrom six other Brassica cultivars were grown on the medium devisedfor Green Comet; shoots were regenerated from two B. oleraceacultivars and two B. napus cultivars, but not from the B. campestriscultivars tested. Brassica oleracea var. italica, Brassica napus, Brassica campestris, seedling root culture, shoot regeneration     

The cDNA Sequence of NS3-Glycoprotein from Brassica campestris and its Homology to Related Proteins

A cDNA clone encoding NS₃-glycoprotein was isolated from stigmasof Brassica campestris. NS-glycoproteins correspond to the SLRI-glycoproteinsof B. oleracea and are highly conserved within the species.These data suggest that the NS-glycoproteins may play a rolein discrimination between species in the fertilization systemof Brassica. (Received September 4, 1992; Accepted November 9, 1992)     

Inter- and intra-genomic homology of the Brassica genomes: implications for their origin and evolution

In order to determine the homologous regions shared by the cultivated Brassica genomes, linkage maps of the diploid cultivated B. rapa (A genome, n = 10), B. nigra (B genome, n = 8) and B. oleracea (C genome, n = 9), were compared. We found intergenomic conserved regions but with extensitve reordering among the genomes. Eighteen linkage groups from all three species could be associated on the basis of homologous segments based on at least three common markers. Intragenomic homologous conservation was also observed for some of the chromosomes of the A, B and C genomes. A possible chromosome phylogenetic pathway based on an ancestral genome of at least five, and no more than seven chromosomes, was drawn from the chromosomal inter-relationships observed. These results demonstrate that extensive duplication and rearrangement have been involved in the formation of the Brassica genomes from a smaller ancestral genome.     

Efficient large-scale development of microsatellites for marker and mapping applications in<Emphasis Type="Italic"> Brassica</Emphasis> crop species

A set of 398 simple sequence repeat markers (SSRs) have been developed and characterised for use with genetic studies of Brassica species. Small-insert (250–900 bp) genomic libraries from Brassica rapa, B. nigra, B. oleracea and B. napus, highly enriched for dinucleotide and trinucleotide SSR motifs, were constructed. Screening the clones with a mixture of oligonucleotide repeat probes revealed positive hybridisation to between 75% and 90% of the clones. Of these, 1,230 were sequenced. Primer pairs were designed for 398 SSR clones, and of these, 270 (67.8%) amplified a PCR product of the expected size in their focal and/or closely related species. A further screen of 138 primers pairs that produced a PCR product in B. napus germplasm found that 86 (62.3%) revealed length polymorphisms within at least one line of a test array representing the four Brassica species. The results of this screen were used to identify 56 SSRs and were combined with 41 SSRs that had previously shown polymorphism between the parents of a B. napus mapping population. These 97 SSR markers were mapped relative to a framework of RFLP markers and detected 136 loci over all 19 linkage groups of the oilseed rape genome.Electronic Supplementary Material Supplementary material is available in the online version of this article at Communicated by O. Savolainen     

A Comparative Study of Certain Seed Isoenzymes in the Ten Chromosome Complex of Brassica Campestris and its Allies

A gel electrophoretic study has been carried out on the seedisoenzymes of n = 10 members of the genus Brassica. Eleven isoenzymesystems have been studied and the distribution of the isoenzymesused to indicate a possible relationship between the major recognizedtaxa. Three basic centres of the complex have been indicated:Indo-European, China and Mediterranean as exemplified by B.campestris (turnip/turnip-rape), B. chinensis and B. tournefortii(wild turnip) respectively.     

S-Locus-Specific Glycoproteins Associated with Self-Incompatibility in Brassica campestris

Specific S-glycoproteins were isolated from three Brassica campestriscultivars homozygous with respect to the S-alleles S₈, S₉ andS₁₂. Amino acid sequences of various peptide fragments of theS-proteins were determined using a gas-phase protein sequencer,and a nearly complete amino acid sequence of the S₈-glycoproteinwas determined on the basis of the revised cDNA sequence ofthe B. oleracea S-specific glycoprotein. The lysyl endopeptidasefragments of S₉ and S₁₂-glycoproteins were aligned in comparisonwith the sequence of the S₈-glycoprotein. Although extensivesequence homology was evident among the three S-glycoproteins,the sequences of the middle part were relatively different fromeach other. The numbers and positions of N-glycosylation alsodiffered among the S-glycoproteins of Brassica species. (Received April 20, 1987; Accepted July 29, 1987)     

Sequence Organization of Simple, Highly Repetitive DNA Elements in Brassica species

The amphidiploid (AACC) nuclear genome of Brassica napus (oil-seedrape) contains c. 5 ? 10⁵ copies of a simple, highly repetitiveDNA element; each repeat is 176 or 177 base pairs long and isdefined by Hind III cutting sites. The diploid (AA) Brassicacampestris (turnip) possesses a very similar repetitive DNA,the consensus sequence of which does not differ from that inB. napus. The 176/177 bp unit consists of three 59 bp sub-units,defined by vestigial EcoRII sites. Analysis of the distributionof variants from consensus in adjacent and non-adjacent unitsprovides evidence for homogenization of sequences by the fixationof independent mutations and for tandem duplication of units.Within units, there is also evidence for inversion and tandemduplication of short (5–8 bp) motifs. Previously published data show that 176/177 base pair repetitiveDNA elements, defined by Hind III cutting sites, are also presentin Sinapis and Raphanus. There is a sequence homology betweenBrassica and Sinapis, and between Brassica and Raphanus, of75%. Sequence homology between Raphanus and Sinapis is 73%. Key words: Repetitive DNA, Brassica, Cruciferae