The suppressive effect of a commercial extract from Durvillaea potatorum and Ascophyllum nodosum on infection of broccoli by Plasmodiophora brassicae.

A sand solution technique demonstrated the capacity for a commercial seaweed extract from Durvillaea potatorum and Ascophyllum nodosum (Seasol Commercial®) to significantly suppress infection of broccoli by Plasmodiophora brassicae. In the primary stages of infection, the extract reduced the number of plasmodia formed in the root hairs by 55 %. Later, in the secondary stages of infection, the extract reduced plasmodia in the root cortical cells by up to 84 %. The suppression of infection was found to be independent of the dilution of the extract applied (1:25 and 1:500). The basis for these results is unlikely to be a nutrient or pH effect since the extract had little impact on these parameters, particularly at the lower dilution (1:200). Rather, we hypothesise that the suppression of infection by the seaweed extract was due to its stimulation of resistance mechanisms in the host, which is possibly related to laminarins in the extract and/or the effect of exogenous growth regulators or undiscovered molecules in the extract disrupting the infection process.

A new interspecific, Gossypium hirsutum x G. barbadense, RIL population: towards a unified consensus linkage map of tetraploid cotton.

We report the development of a new interspecific cotton recombinant inbred line (RIL) population of 140 lines deriving from an interspecific cross between Gossypium hirsutum (Gh) and G. barbadense (Gb), using the same two parents that have served for the construction of a BC(1) map and for the marker-assisted backcross selection program underway at CIRAD. Two marker systems, microsatellites and AFLPs, were used. An important feature of the RIL population was its marked segregation distortion with a genome-wide bias to Gh alleles (parental genome ratio is 71/29). The RIL map displays an excellent colinearity with the BC(1) map, although it is severely contracted in terms of map size. Existence of 255 loci in common (between 6 and 14 per chromosome) allowed the integration of the two data sets. A consensus BC(1)-RIL map based upon 215 individuals (75 BC1 + 140 RIL) was built. It consisted of 1,745 loci, spanned 3,637 cM, intermediate between the sizes of the two component maps, and constituted a solid framework to cross align cotton maps using common markers. The new RIL population will be further exploited for fiber property QTL mapping and eQTL mapping.

Morphology of rsw1, a cellulose-deficient mutant of Arabidopsis thaliana.

The rsw1 mutant of Arabidopsis thaliana is mutated in a gene encoding a cellulose synthase catalytic subunit. Mutant seedlings produce almost as much cellulose as the wild type at 21 degrees C but only about half as much as the wild type at 31 degrees C. We used this conditional phenotype to investigate how reduced cellulose production affects growth and morphogenesis in various parts of the plant. Roots swell in all tissues at 31 degrees C, and temperature changes can repeatedly switch them between swollen and slender growth patterns. Dark-grown hypocotyls also swell, whereas cotyledons and rosette leaf blades are smaller, their surfaces are more irregular and their petioles shorter. Leaf trichomes swell and branch abnormally. Plants readily initiate inflorescences at 31 degrees C which have shorter but not fatter bolts and stomata which bulge above the uneven surface of internodes. Bolts carry the normal number of flowers, but their stigmas protrude beyond the shortened sepals and petals. Anthers dehisce normally, but self-fertilisation is reduced because the stigma is well above the anthers. Anther filaments are short and show a crumpled surface. Viable pollen develops, but female reproductive competence and postpollination development are severely impaired. We conclude that the RSW1 gene is important for cellulose synthesis in many parts of the plant and that reduced cellulose synthesis suppresses organ expansion rather than organ initiation, causes radial swelling only in the root and hypocotyl, but makes the surfaces of many organs uneven. We discuss some possible reasons to explain why different organs vary in their responses. The morphological changes suggest that RSW1 contributes cellulose to primary walls but do not yet exclude a role during secondary-wall deposition.

Temperature-sensitive alleles of RSW2 link the KORRIGAN endo-1,4-beta-glucanase to cellulose synthesis and cytokinesis in Arabidopsis.

An 8.5-kb cosmid containing the KORRIGAN gene complements the cellulose-deficient rsw2-1 mutant of Arabidopsis. Three temperature-sensitive alleles of rsw2 show single amino acid mutations in the putative endo-1,4-beta-glucanase encoded by KOR. The F1 from crosses between kor-1 and rsw2 alleles shows a weak, temperature-sensitive root phenotype. The shoots of rsw2-1 seedlings produce less cellulose and accumulate a short chain, readily extractable glucan resembling that reported for rsw1 (which is defective in a putative glycosyltransferase required for cellulose synthesis). The double mutant (rsw2-1 rsw1) shows further reductions in cellulose production relative to both single mutants, constitutively slow root growth, and enhanced temperature-sensitive responses that are typically more severe than in either single mutant. Abnormal cytokinesis and severely reduced birefringent retardation in elongating root cell walls of rsw2 link the enzyme to cellulose production for primary cell walls and probably cell plates. The Rsw2(-) phenotype generally resembles the Kor(-) and cellulose-deficient Rsw1(-) phenotypes, but anther dehiscence is impaired in Rsw2-1(-). The findings link a second putative enzyme activity to cellulose synthesis in primary cell walls of Arabidopsis and further increases the parallels to cellulose synthesis in Agrobacterium tumefaciens where the celA and celC genes are required and encode a putative glycosyltransferase and an endo-1,4-beta-glucanase related to RSW1 and KOR, respectively.

Molecular analysis of cellulose biosynthesis in Arabidopsis.

Cellulose, an abundant, crystalline polysaccharide, is central to plant morphogenesis and to many industries. Chemical and ultrastructural analyses together with map-based cloning indicate that the RSW1 locus of Arabidopsis encodes the catalytic subunit of cellulose synthase. The cloned gene complements the rsw1 mutant whose temperature-sensitive allele is changed in one amino acid. The mutant allele causes a specific reduction in cellulose synthesis, accumulation of noncrystalline beta-1,4-glucan, disassembly of cellulose synthase, and widespread morphological abnormalities. Microfibril crystallization may require proper assembly of the RSW1 gene product into synthase complexes whereas glucan biosynthesis per se does not.

In Trifolium subterraneum, chalcone synthase is encoded by a multigene family.

Chalcone synthase (CHS) catalyzes the first and key regulatory step in flavonoid biosynthesis. We report the existence and characterization of a CHS multigene family present in Trifolium subterraneum L. cultivar Karridale. The CHS family consists of at least four members, which are tightly clustered in a 15-kb region. The complete sequences of two of these genes (CHS1 and CHS2) are presented. The putative promoters of these genes have sequences which are homologous to those known, or implicated, in regulation of the expression of phenylpropanoid-encoding genes.

Characterization of a phenylalanine ammonia-lyase multigene family in Trifolium subterraneum.

The enzyme phenylalanine ammonia-lyase (PAL) was found to be encoded by a small gene family in the legume Trifolium subterraneum (subterranean clover). At least three of the family members are tightly clustered within approx. 20 kb of DNA. Sequencing of one of the genes established that it possesses two exons, the position of the single intron being identical to that found for PAL genes from other plants. The PAL protein consists of 725 amino acids, as deduced from the nucleotide sequence.