Seed characteristics and fatty acid composition of castor (<i>Ricinus communis</i> L.) varieties in Northeast China

Oil content and fatty acid composition were investigated on 12 castor varieties and strains by using the soxhlet extraction method and capillary gas chromatography. This was made to provide a reference and theoretical basis for castorbean breeding with high oil content, determine variability of seed compounds for breeding purposes, and broaden chemical material choices. Results revealed that crude fat percentage in seeds ranged from 18.91 to 35.84% with an average of 25.91%; the absolute content of ricinoleic acid varied between 171.65 g/kg and 314.03 g/kg with an average of 222.43 g/kg, and kernel crude fat percentage was between 24.28 and 46.97% with an average of 34.30%. All these study variables were highest in the 2129 strain. The percentage of ricinoleic acid in crude fat was between 83.85 to 87.62%, and the highest value was found in the zhebi4 accession. The other fatty acids appeared in small concentrations, and showed small amplitude: 1.12 to 1.61%, 1.21 to 1.61%, 3.53 to 4.80%, 5.35 to 6.38%, 0.52 to 0.79%, 0.05 to 0.08% and 0.43 to 0.55%, for palmitic, stearic, oleic, linolic, linolenic, arachidic, and arachidonic acids, respectively. One hundred seed weight was determined for each accession. One hundred seed weight ranged from 25.7 g to 34.0 g with an average of 29.9 g. There was a significant correlation between seed weight and oil content, but the correlation value was low (r=0.51). Cluster analysis by SSPS based on the content of fatty acid composition revealed that the accessions were divided into three independent clusters. These findings will clearly provide useful information for further research in breeding and utilization of castor oil.     

<Emphasis Type="Italic">Brassica</Emphasis> biotechnology: Progress in cellular and molecular biology

Summary Considerable progress has been accomplished in the cellular and molecular biology of Brassica species in the past few years. Plant regeneration has been increasingly optimized via organogenesis and somatic embryogenesis using various explants; with tissue culture improvements focusing on factors such as age of the explant, genotype, and media additives. The production of haploids and doubled haploids using microspores has accelerated the production of homozygous lines in the Brassica species. Somatic cell fusion has facilitated the development of interspecific and intergeneric hybrids in the sexually incompatible species of Brassica. Crop improvement using somaclonal variation has also been achieved. The use of molecular markers in marker-assisted selection and breeding, transformation technology for the introduction of desirable traits, and a comparative analysis of these as well as their future prospects are important parts of the current research that is reviewed.     

A cephalosporin C acetylhydrolase is present in the cultures of Nocardia lactamdurans

Two protein bands with strong esterase activity are present in broths of Nocardia lactamdurans MA4213 cultures. One of them shows cephalosporin C acetylhydrolase (CAH) activity. This activity is maximal at 48 h of growth and shows a pattern of regulation slightly different from that of cephamycin production in medium supplemented with glucose (166 mM), glycerol (326 mM) or ammonium chloride (60 mM). The CAH activity was purified to homogeneity by DEAE-Sepharose ion-exchange, Sephadex G-75 gel filtration, and phenyl-Sepharose hydrophobic interaction chromatography. It showed a molecular mass of 72,100 Da. The N-terminus of the protein was determined and showed the amino acid sequence GGAAPGGPGAHPLWLPAGKD. The enzyme showed K _m values of 7.0 mM and 8.3 mM for cephalosporin C and 7-aminocephalosporanic acid respectively but was not active on cephamycin C. Received: 17 December 1999 / Received revision: 22 February 2000 / Accepted: 25 February 2000     

Overexpression and lack of degradation of thaumatin in an aspergillopepsin A‐defective mutant of Aspergillus awamori containing an insertion in the pep A gene

A gene encoding the sweet-tasting protein thaumatin (tha) with optimized codon usage was expressed in Aspergillus awamori. Mutants of A. awamori with reduced proteolytic activity were isolated. One of these mutants, named lpr66, contained an insertion of about 200 bp in the pepA gene, resulting in an inactive aspergillopepsin A. In vitro thaumatin degradation tests confirmed that culture broths of mutant lpr66 showed only a small thaumatin-degrading activity. A. awamori lpr66 has been used as host strain for thaumatin expression cassettes containing the tha gene under the control of either the cahB (cephalosporin acetylhydrolase) promoter of Acremonium chrysogenum or the gdhA (glutamate dehydrogenase) promoter of Aspergillus awamori. Residual proteolytic activities were repressed by using a mixture of glucose and sucrose as carbon sources and l-asparagine as nitrogen source. Degradation of thaumatin by acidic proteases was prevented by maintaining the pH value at 6.2 in the fermentor. Expression of cassettes containing the gdhA promoter was optimal in ammonium sulfate as nitrogen source, whereas transformants expressing the tha gene from the cahB promoter yielded higher thaumatin levels using l-asparagine as nitrogen source. Under optimal fermentation conditions, yields of 105 mg thaumatin/l were obtained, thus making this fermentation a process of industrial interest. Received: 27 March 2000 / Received revision: 14 June 2000 / Accepted: 18 June 2000     

Effects of Trichothecene Production on the Plant Defense Response and Fungal Physiology: Overexpression of the Trichoderma arundinaceum tri4 Gene in T. harzianum

Trichothecenes are fungal sesquiterpenoid compounds, the majority of which have phytotoxic activity. They contaminate food and feed stocks, resulting in potential harm to animals and human beings. Trichoderma brevicompactum and T. arundinaceum produce trichodermin and harzianum A (HA), respectively, two trichothecenes that show different bioactive properties. Both compounds have remarkable antibiotic and cytotoxic activities, but in addition, trichodermin is highly phytotoxic, while HA lacks this activity when analyzed in vivo. Analysis of Fusarium trichothecene intermediates led to the conclusion that most of them, with the exception of the hydrocarbon precursor trichodiene (TD), have a detectable phytotoxic activity which is not directly related to the structural complexity of the intermediate. In the present work, the HA intermediate 12,13-epoxytrichothec-9-ene (EPT) was produced by expression of the T. arundinaceum tri4 gene in a transgenic T. harzianum strain that already produces TD after transformation with the T. arundinaceum tri5 gene. Purified EPT did not show antifungal or phytotoxic activity, while purified HA showed both antifungal and phytotoxic activities. However, the use of the transgenic T. harzianum tri4 strain induced a downregulation of defense-related genes in tomato plants and also downregulated plant genes involved in fungal root colonization. The production of EPT by the transgenic tri4 strain raised levels of erg1 expression and reduced squalene accumulation while not affecting levels of ergosterol. Together, these results indicate the complex interactions among trichothecene intermediates, fungal antagonists, and host plants.     

Multipartite symbioses among fungi, mites, nematodes, and the spruce beetle, Dendroctonus rufipennis

The spruce beetle, Dendroctonus rufipennis, is an eruptive forest pest of significant economic and ecological importance. D. rufipennis has symbiotic associations with a number of microorganisms, especially the ophiostomatoid fungus Leptographium abietinum. The nature of this interaction is only partially understood. Additionally, mite and nematode associates can mediate bark beetle-fungal interactions, but this has not yet been studied for spruce beetles. In this study, we found eight mite species associated with spruce beetles: Tarsonemus ips, T. endophloeus, Histiogaster arborsignis, Dendrolaelaps quadrisetus, Proctolaelaps hytricoides, Trichouropoda alascae, T. n. sp. nr dalarenaensis, and Urobovella n. sp 767. The most prevalent species was H. arborsignis. In addition, 75% of beetles examined carried nematodes, with six species represented. These included a new species of Parasitorhabditis, Ektaphelenchus obtusus, Bursaphelenchus n. sp. 727, Aphelenchoides n. sp., Panagrolaimus sp., and Mykoletzkya ruminis. H. arborsignis showed strong feeding and oviposition preferences for L. abietinum among four fungal species tested in laboratory assays. Information on our attempts to culture the various nematode species collected from D. rufipennis is also provided. Bursaphelenchus were cultured from D. rufipennis nematangia plated on agar containing L. abietinum but not sterile agar. Thus, L. abietinum plays an important role in these gallery communities, affecting the tree-killing bark beetle, its phoretic mites, and nematodes. These data add to our understanding of bark beetle-microorganism interactions.     

Cloning and characterization of the erg1 gene of Trichoderma harzianum: effect of the erg1 silencing on ergosterol biosynthesis and resistance to terbinafine

Trichoderma species are commonly used as biocontrol agents of different plant-pathogenic fungi. Terpene compounds are involved in the biocontrol process due to their antifungal properties (e.g., ergokonins and viridins) but additionally their structural function in the cell membranes (ergosterol) is essential. We report here the characterization of the T. harzianum erg1 gene, encoding a squalene epoxidase, a key enzyme in the biosynthesis of triterpene derivatives such as ergosterol. In T. harzianum the partial silencing of the erg1 gene gave rise to transformants with a higher level of sensitivity to terbinafine, an antifungal compound that acts specifically over the squalene epoxidase activity. In addition, these silenced transformants produced lower levels of ergosterol than the wild type strain. Finally, the silencing of the erg1 gene resulted in an increase in the expression level of the erg7 gene that encodes the oxidosqualene lanosterol-cyclase, another enzyme of the terpene biosynthesis pathway.