Plant Molecular Genetics Research Articles

From the mechanisms of genetic transposition to the functional genomics

Pioneer works on studying molecular mechanisms of mutagenesis were published in the journal Genetika in the 1960s. In the laboratory of S.I. Alikhanian, studies on molecular mechanisms of genetic transposition were initiated in the late 1960s on the model of bacteriophage transposon Mu (Mutator). Parallel to these studies conducted in the laboratory of plant molecular genetics (Institute of Molecular Genetics, Academy of Sciences of the USSR), which was later named the Laboratory of Functional Genomics (Vavilov Institute of General Genetics, Russian Academy of Sciences), studies on transposition of Ti-plasmid T-DNA of Agrobacterium tumefaciens and works on construction of transgenic plants began in this laboratory. Transgenic plants with the expressed bacterial genes provided a model for the functional genomics. This topic is considered here in detail.

Identification of Glycine max Genes Expressed in Response to Soybean mosaic virus Infection

School of Agricultural Biotechnology and Center for Plant Molecular Genetics and Breeding Research, Seoul NationalUniversity, Seoul 151-921, Korea(Received on November 3, 2004; Accepted on December 29, 2004)Identification of host genes involved in disease prog-resses and/or defense responses is one of the mostcritical steps leading to the elucidation of diseaseresistance mechanisms in plants. Soybean mosaic virus(SMV) is one of the most prevalent pathogen of soybean(Glycine max). Although the soybeans are placed one ofmany important crops, relatively little is known aboutdefense mechanism. In order to obtain host genesinvolved in SMV disease progress and host defenseespecially for virus resistance, two different cloningstrategies (DD RT-PCR and Subtractive hybridization)were employed to identify pathogenesis- and defense-related genes (PRs and DRs) from susceptible (Geum-jeong 1) and resistant (Geumjeong 2) cultivars againstSMV strain G7H. Using these approaches, we obtained570 genes that expressed differentially during SMVinfection processes. Based upon sequence analyses,differentially expressed host genes were classified intofive groups, i.e. metabolism, genetic information pro-cessing, environmental information processing, cellularprocesses and unclassified group. A total of 11 differ-entially expressed genes including protein kinase, tran-scription factor, other potential signaling componentsand resistant-like gene involved in host defense responsewere selected to further characterize and determineexpression profiles of each selected gene. Functionalcharacterization of these genes will likely facilitate theelucidation of defense signal transduction and biologicalfunction in SMV-infected soybean plants.Keywords : defense, differential expression, EST analysis,pathogenesis, SMVPlants have developed complicated defense mechanismduring evolution to resist the harmful pathogens they haveencountered. The interaction between a plant and a pathogenoften complies with gene-for-gene model involving plantresistance (R) gene and corresponding pathogen avirulence(Avr) genes (Flor, 1971). These involve changes in ioninflux, generation of reactive oxygen species, production ofpathogenesis-related (PR) proteins and phytoalexins, andreinforcement of the cell wall (Dixon and Lamb, 1990;Dixon et al., 1994). Induction of defense response involvesan initial signal that indicating invasion by a pathogen,followed by transduction of signals activating variousdefense genes (Yang et al., 1997). When plants recognizethe products of Avr genes directly or indirectly from apathogen, the defense-related gene (DR) expression isinduced and the result is resistance to pathogen attack(Keen, 1990; Staskawicz, 2001). However, when plantslack R genes or their products, the plant displaycharacteristics of disease. Based on the advancements ofhost-pathogen interaction at the molecular level, diseaseresistance genes are useful for disease control strategies inagriculture.Soybean mosaic virus (SMV), a member of the genusPotyvirus (Mayo and Pringle, 1998), is one of the mosteconomically important viruses of soybean [Glycine max(L.) Merr]. It cause mosaic and severe necrosis in manysoybean cultivars, and is present in all major soybean-growing areas, where it results in significant reductions inyield and quality. Since the first description of seven groupsof SMV strains (G1-G7) based on the symptoms developedon test cultivars, viral strain variation and the resistancepatterns of different soybean cultivars have been studiedextensively (Cho and Goodman, 1979, 1982; Gunduz et al.,2001, 2002; Lim, 1985). Presence of many SMV strains hasbeen reported and the continual variation is generallybelieved to happen in soybean fields in Korea (Cho et al.,1983). Recently, a new SMV strain, G7H, causing mosaicand necrosis on most recommended soybean cultivars thatare resistant to previously prevalent SMV strains, G5 andG5H, was found in Korea (Kim et al., 2003; Lim et al.,2003).To date, the majority of research in defense signaltransduction has been performed on several model speciessuch as tobacco and Arabidopsis, whereas in economicallyimportant soybean crop, our understanding of the signalingmechanisms leading to disease resistance is largely unknown(Piffanelli et al., 1999). Most characterized R genes can be

Molecular Population Genetics and the Search for Adaptive Evolution in Plants

The first papers on plant molecular population genetics were published approximately 10 years ago. Since that time, well over 50 additional studies of plant nucleotide polymorphism have been published, and many of these studies focused on detecting the signature of balancing or positive selection at a locus. In this review, we discuss some of the theoretical and statistical issues surrounding the detection of selection, with focus on plant populations, and we also summarize the empirical plant molecular population genetics literature. At face value, the literature suggests that a history of balancing or positive selection in plant genes is rampant. In two well-studied taxa (maize and Arabidopsis) over 20% of studied genes have been interpreted as containing the signature of selection. We argue that this is probably an overstatement of the prevalence of natural selection in plant genomes, for two reasons. First, demographic effects are difficult to incorporate and have generally not been well integrated into the plant population genetics literature. Second, the genes studied to date are not a random sample, so selected genes may be overrepresented. The next generation of studies in plant molecular population genetics requires additional sampling of local populations, explicit comparisons among loci, and improved theoretical methods to control for demography. Eventually, candidate loci should be confirmed by explicit consideration of phenotypic effects.

The “Prochaska” Microtiter Plate Bioassay for Inducers of NQO1

This chapter provides details of ‘‘Prochaska’’ microtiter plate bioassay for inducers of NQO1. The microtiter plate bioassay for NQO1 was first developed in the laboratory by Hans Prochaska as a rapid and direct assay of quinone reductase (NQO1; QR; DT-diaphorase) activity in cultured cells, suitable for identifying, purifying, and determining the potency of inducers of this detoxication enzyme. The Prochaska microtiter plate bioassay has been exceptionally useful for identifying and isolating inducers of NQO1 from natural sources, and in guiding the synthesis of more potent analogs of isothiocyanates, dithiolethiones, and curcuminoids. It has also been used to survey screen, and compare the potency of various plant sources in both plant breeding and molecular genetics applications. An overview of Prochaska bioassay protocol is presented in this chapter, elaborating the technical improvements of the assay. Novel findings made with the Prochaska bioassay are also described in the chapter. The chapter elaborates the and limitations of the Prochaska bioassay. Test compound matrix, dosing, and bioassay variability are also analyzed in the chapter.

Frontiers of plant cell biology: signals and pathways, system-based approaches 22nd Symposium in Plant Biology (University of California--Riverside).

The symposium ''Frontiers of Plant Cell Biology: Signals and Pathways, Systems-Based Approaches'' was held January 15-18, 2003 at the Riverside Convention Center in Riverside, California. The host organization for the symposium was the Center for Plant Cell Biology (CEPCEB) at the University of California, Riverside (UCR). The meeting, focusing on systems-based approaches to plant cell biology research, was the first of this kind in the field of plant biology. The speakers and nearly 100 posters placed emphasis on recent developments in plant cellular biology and molecular genetics, particularly those employing emerging genomic tools, thereby sharing the most current knowledge in the field and stimulating future advances. In attendance were many well-established scientists and young investigators who approach plant cell biology from different but complementary conceptual and technical perspectives. Indeed, many disciplines are converging in the field of cell biology, producing synergies that will enable plant scientists to determine the function of gene products in the context of living cells in whole organisms. New, cross-disciplinary collaborations, as well as the involvement of computer scientists and chemists in plant biology research, are likely additional outcomes of the symposium. The program included 39 invited session speakers and workshop/panel speakers. Sessions were convened on the following themes: Cell-Cell Communication; Protein Trafficking; Cell Surface, Extracellular Matrix and Cell Wall; Signal Transduction; Signal Transduction and Proteosome; and Systems-Based Approaches to Plant Cell Biology. Workshops on Chemical Genetics and Visual Microscopy were also presented. Abstracts from each of the speaker presentations, as well as the posters presented at the meeting were published in a program booklet given to the 239 faculty members, researchers, postdoctoral scientists and graduate students in attendance. The booklet thus serves as a reference for symposium attendees to locate additional information about a topic of their particular interest and to contact other investigators. In addition, an article reviewing the symposium by science writer Peter V. Minorsky appeared in the June 2003 issue of Plant Physiology, a special issue devoted to systems-based approaches in the study of the model plant Arabidopsis (article submitted as part of this Final Technical Report).

The impact of the changing fatty acid profile of fats on diet assessment and health

We are transiting a time in which advances in plant molecular biology and genetics have resulted in a remarkable array of oils and fats that have a modified fatty acid profile, compared to the parent stock variety. The rapid evolution of modified oils and fats poses challenges for keeping nutrient databases current as well as deciphering what new fats/oils are used, from the standpoint of diet assessment and, in particular, collecting accurate dietary data. The expanding array of modified fats and oils has been designed to have greater shelf stability due to an increase in monounsaturated fat and enhanced health benefits due to a reduction in trans fat. The effects of oilseeds reduced in alpha-linolenic acid (ALA) on health outcomes need to be evaluated since diets rich in ALA have many health benefits.

Prospects for improving nitrogen use efficiency: Insights given by 15N-labelling experiments

In higher plants, recent advances in plant molecular genetics, combined with modern physiological and biochemical studies, have expanded our understanding of the regulatory mechanisms controlling the primary steps of inorganic nitrogen assimilation and the subsequent biochemical pathways involved in nitrogen supply and recycling for higher plant metabolism, growth and development. In this presentation, we describe improvements in our understanding of the molecular controls of nitrogen assimilation through the use of transgenic plants and the study of genetic variability in model and crop species. To illustrate this research programme, the physiological impact of modified gene expression, using either transgenic plants or different genotypes, was studied using 15N-labelling experiments in order to monitor the influx of nitrate or ammonia and its subsequent incorporation into amino acids.

Exposing Students and Teachers to Plant Molecular Genetics with Short-Term Barley Gene Mapping Projects

Many universities sponsor science research programs during the summer to provide hands-on laboratory experience to high school students and teachers. Our objective was to design a project that exposes the students to the full range of research, from developing and testing a hypothesis through presenting research results. The project goal is to demonstrate mapping of morphological traits using simple sequence repeat (SSR) markers. Students learn how to evaluate plants for morphological marker traits, extract DNA, conduct PCR (polymerase chain reaction) and gel electrophoresis, evaluate results, and conduct linkage analysis between traits and markers. Mapping data are presented for five of the barley (Hordeum vulgare L.) traits analyzed since 1997. The genes bracteatum (bra-a.001) and short awn (lks.o) were located on the short arm of chromosome 7H, the glossy phenotype conferred by cer-zv.268 was located near the centromere of chromosome 4H, fragile stem (fst2.b) was located near the centromere of chromosome 1H, and intermedium spike (int-h.42) was located on the short arm of chromosome 5H. The high school teachers have used their experiences to help teach genetics and molecular biology, and several have included the projects in their portfolio for an advanced degree. Many students have used their projects for science fairs and their experience to help obtain part-time jobs during their undergraduate studies. These experiments provide the necessary components of a successful research project for short-term programs and provide meaningful data to advance our ongoing projects, benefitting both the students and our laboratory.

Characterization of the Selaginella remotifolia MADS-box gene

Recent progress in plant molecular genetics has revealed that floral organ development is regulated by several homeotic selector genes, most of which belong to the MADS-box gene family. Here we report on SrMADS1,a MIKC(c)-type MADS-box gene from Selaginella, a spikemoss belonging to the lycophytes. SrMADS1 phylogenetically forms a monophyletic clade with genes of the LAMB2 group, which are MIKC(c) genes of the clubmoss Lycopodium, and is expressed in whole sporophytic tissues except roots and rhizophores. Our results and the previous report on Lycopodium MIKC(c) genes suggest that the ancestral MIKC(c )gene of primitive dichotomous plants in the early Devonian was involved in the development of basic sporophytic tissues such as shoot, stem, and sporangium.

Impact of biotechnology on agriculture in China

China, the most populous developingcountry in the world, facing greatpressures regarding population growth,environmental deterioration and resourcedepletion, has ranked plant biology as one of its highest priorities since theearly 1950s, especially in the area ofagricultural development. Since plantbiotechnology became available, it hasdeveloped rapidly in agriculture-relatedplant biology research.Two crucial innovations – tissue andanther culture and DNArecombination –marked the emergence of plantbiotechnology onto the world stage in the 1960s and 1970s. Unfortunately,China was immersed in the CulturalRevolution during that period, and plantscientists were not allowed to do any basic research, although some researchrelated to crop breeding was permitted.For ~20 years, plant tissue culture andanther culture were among the mostactive research areas in plant science inChina because it was believed to be anefficient tool for plant breeding.Many talented young scientists at thattime devoted their careers to developingand improving the technology [1]. To date, >700 plant species have beencultivated successfully via tissue and/oranther culture [2], and >20 new agronomicvarieties have been obtained using thesetechniques [1].DNArecombination techniques were not introduced into China until theearly 1980s because of a lack of facilities,reagents and personnel with propertraining. Before that, isozyme analysiswith electrophoresis was popular, in part,because people believed that with thisinexpensive and relatively simpletechnique it was possible to investigategenetic as well as physiological changesin breeding and the regulation of plantgrowth. Also, because hybrid rice wasmassively used during that time, therewere demands for molecular markers for screening and quality control of hybrid seeds. During the mid-1980s, the first batch ofvisiting scholars sent to Western countriesafter the Cultural Revolution introducedDNArecombination techniques intoChina. Scientists, mainly those based inthe institutes of the Chinese Academy ofSciences (CAS) such as the Institute ofMicrobiology (Beijing), the ShanghaiInstitute of Plant Physiology, theShanghai Institute of Biochemistry andthe Institute of Genetics (Beijing), startedto work with plasmids, viruses andAgrobacteria. Over about five years, the creation of the National Laboratory of Plant Molecular Genetics (Shanghai)laid the foundation for plant molecularbiological research in China. Onenoteworthy invention from the Institute of Biochemistry by Guang-Yu Zhou, is the technique of transforming foreigngenes via the pollen tube pathway [3].Although this approach has beencontroversial since it was proposed, many transformations via this approachhave proven successful at the molecularlevel in crops such as cotton, rice, wheat,maize and soybean [4].After the introduction of DNArecombination techniques into China,Chinese scientists eagerly undertookprotoplast manipulation in the mid-1980s.More than 50 plant species have beengenetically engineered in China to date [1]. Although research into protoplast manipulation declined after the establishment of plant transformationwith

Grasses and gall midges: plant defense and insect adaptation.

The interactions of two economically important gall midge species, the rice gall midge and the Hessian fly, with their host plants, rice and wheat, respectively, are characterized by plant defense via R genes and insect adaptation via avr genes. The interaction of a third gall midge species, the orange wheat blossom midge, with wheat defense R genes has not yet exhibited insect adaptation. Because of the simple genetics underlying important aspects of these gall midge-grass interactions, a unique opportunity exists for integrating plant and insect molecular genetics with coevolutionary ecology. We present an overview of some genetic, physiological, behavioral, and ecological studies that will contribute to this integration and point to areas in need of study.

PCR‐genotyping of barley seedlings using DNA samples from tissue prints

AbstractScreening of large numbers of plant samples with polymerase chain reaction (PCR)‐based techniques is a central task in marker‐assisted plant breeding and molecular genetics. While PCR and electrophoresis have been streamlined as a result of high‐throughput analysis methods, the collection of plant material and its processing to make it accessible for reliable amplification reactions remain labour‐intensive, costly and not amenable to automation. As an alternative to traditional extraction protocols, DNA can be bound to specially treated paper (FTA‐paper). For PCR analysis, instead of being added as a liquid suspension, DNA is then released from small paper disks that are immersed into the reaction mixture. In the present study, several parameters were investigated for the successful amplification of various single‐copy sequence characterized amplified region markers from barley tissue prints. The results show that the FTA‐paper approach offers a quick, reliable and low‐cost alternative for PCR screening even in a crop plant with a large genome.

Genome walking with consensus primers: application to the large single copy region of chloroplast DNA

AbstractConserved chloroplast (cp) DNA primer pairs are useful in plant molecular genetics, evolution and ecology. We have designed 20 conserved cpDNA primer pairs that, in combination with 18 previously described ones, amplify overlapping fragments (mean size of 2.5 kb) spanning the large single copy (LSC) region from Eudicots. These 38 primer pairs as well as eight primer pairs flanking cpDNA microsatellites were tested on 20 plant species belonging to 13 families. At least 79% and up to 100% of the LSC (> 86 kb) can be amplified. Many primer pairs are robust and work with all species.

The bases of crown gall tumorigenesis.

The nine decades since Smith and Townsend demonstrated that Agrobacterium tumefaciens causes plant tumors (95) have been marked by a series of surprises. Among the most important of these was the report in 1958 that these tumors could be excised and propagated in vitro without exogenous plant hormones (7). Equally important were a series of reports beginning about the same time that tumors released compounds that agrobacteria could use as nutrients (24). Perhaps the most exciting discoveries, reported in the 1970s and 1980s, were that tumorigenesis required the transfer of fragments of oncogenic DNA to infected plant cells (10), that this process evolved from a conjugal transfer system (99), and that the genes that direct this process are expressed in response to host-released chemical signals (47). This DNA transfer process has become a cornerstone of plant molecular genetics. The genus Agrobacterium also has provided excellent models for several aspects of host-pathogen interactions, including intercellular transport of macromolecules (11), bacterial detection of host organisms (47), targeting of proteins to plant cell nuclei (3), and interbacterial chemical signaling via autoinducer-type pheromones (120).

Transactivation of a target gene through feedforward loop activation in plants.

The expression of many genes encoding transcriptional activators in prokaryotes and eukaryotes is upregulated through positive feedback activation. During positive feedback activation, a transcriptional activator binds to its own promoter and thus increases its own expression as well as the expression of its target genes. In the simplest case, increased levels of the transcriptional activator can be directly correlated with increased expression of its target genes. In this study, we present a gene expression system, designated feedforward loop (FFL) system, which makes use of this kind of positive feedback regulation for the expression of plant transgenes. We show in transient and stable transformation experiments that such a system is functional and that it can be used to obtain high level gene expression in plants. We also provide evidence that the transgene is ubiquitously expressed when using the FFL system. Finally, we discuss the possibilities of using FFL gene expression systems for applications in plant molecular genetics and biotechnology.

Chemicals from biotechnology: molecular plant genetics will challenge the chemical and the fermentation industry.

Industrial biotechnology has evolved as a significant manufacturing tool for products like fuel-grade ethanol, organic acids and bulk amino acids, but most items are still speciality products for food and pharmaceutical applications. Current development projects within the chemical industry, including lactic acid and 1,3-propanediol based polymers and plastics, indicate that new biotechnological processes and products may soon approach the market place, clearly targeted at the leading petrochemical bulk outlets. This is flanked by a strategic shift by the major chemical companies in to "life sciences"-pharmaceuticals, agrochemicals and the seed business as well as biotech fine chemicals. The recent tremendous achievements in molecular plant genetics and transgenic crop breeding will boost agrobiotechnology, agriculture and renewable raw materials as compelling projects for chemistry and biotechnology. New plant-based production routes may challenge established chemical and biochemical domains, but at the same time open new horizons to valuable feedstocks, intermediates and end-products.

A complete BAC-based physical map of the Arabidopsis thaliana genome.

Arabidopsis thaliana is a small flowering plant that serves as the major model system in plant molecular genetics. The efforts of many scientists have produced genetic maps that provide extensive coverage of the genome (http://genome-www. stanford.edu/Arabidopsis/maps.html). Recently, detailed YAC, BAC, P1 and cosmid-based physical maps (that is, representations of genomic regions as sets of overlapping clones of corresponding libraries) have been established that extend over wide genomic areas ranging from several hundreds of kilobases to entire chromosomes. These maps provide an entry to gain deeper insight into the A. thaliana genome structure. A. thaliana has been chosen as the subject of the first large-scale project intended to determine the full genome sequence of a plant. This sequencing project, together with the increasing interest in map-based gene cloning, has highlighted the requirement for a complete and accurate physical map of this plant species. To supply the scientific community with a high-quality resource, we present here a complete physical map of A. thaliana using essentially the IGF BAC library. The map consists of 27 contigs that cover the entire genome, except for the presumptive centromeric regions, nucleolar organization regions (NOR) and telomeric areas. This is the first reported map of a complex organism based entirely on BAC clones and it represents the most homogeneous and complete physical map established to date for any plant genome. Furthermore, the analysis performed here serves as a model for an efficient physical mapping procedure using BAC clones that can be applied to other complex genomes.

Putative Components of the Arabidopsis Circadian Clock

Although plant circadian rhythms have been described as far back as Antiquity, the molecular basis of the plant biological clock is still unknown. The reasons for this lag behind Neurospora, Drosophila and now mammalian systems has primarily been the lack of suitable genetic systems and of easily screenable phenotypes for the isolation of mutants with defective circadian rhythms. In the past few years, the small, fast growing weed Arabidopsis thaliana has become a widely used system for plant molecular genetics, and extensive genome mapping and sequencing programs have greatly facilitated gene isolation. New, non-invasive assays for plant circadian rhythms have been developed, that made the isolation of clock mutants possible. Defective circadian rhythms have been shown to correlate with visible phenotypes, such as aberrant flowering times, and these phenotypes have been used successfully in the isolation of mutants with altered clock function. As a consequence plant circadian biology is now experiencing a renewed interest, due to the isolation of a number of putative components of the circadian regulatory system. The first break-through was from Steve Kay and colleagues, who fused the promoter of the circadian-regulated CAB (chlorophyll a/b-binding protein) gene to a firefly luciferase reporter gene. Transgenic Arabidopsis plants carrying this transgene exhibited rhythmic luminescence when supplied with the substrate luciferin. The luminescence rhythm was monitored using a photon-counting camera. This non-invasive assay was used to isolate mutants with altered rhythmicity (Millar et al., 1995a). One mutant isolated by this method, toc1 (Timing of CAB-1) has been extensively characterized. The toc1 mutation was shown to shorten the period of a variety of circadian rhythms, including CAB (Millar et al., 1995a) and

Molecular genetics of Physcomitrella

In the first few decades of this century, scientists working on the genetics of bryophytes mosses and liverworts were at the forefront of genetical research: Allen (1917) was the first to describe sex chromosomes in plants, Heitz (1928) demonstrated the continuity of chromosomes during the mitotic cell cycle, and Knapp (1936) employed X-ray mutagenesis for genetical research. Although the beginning of plant molecular genetics was marked by ultraviolet light (UV) mutagenesis of the liverwort Sphaerocarpos donellii, which demonstrated DNA as the molecular basis of inheritance (Knapp et al. 1939), bryophytes have only been of marginal interest for modern molecular biologists. The outstanding exception is the liverwort Marchantia polymorpha, which was first subjected to genetical analysis with intensive mutation and cross-breeding experiments (Burgeff 1943). It is now well known because complete nucleotide sequences have been determined for its plastid and its mitochondrial DNA (Ohyama et al. 1986; Oda et al. 1992). Non-mendelian inheritance was first postulated by von Wettstein (1928) who analysed Funaria hygrometri ca and related mosses. Likewise, von Wettstein (1924) recognised the great potential of the haploid moss protonemata for genetically dissecting differentiation processes. These analyses started with mutant induction in Physcomitrium piriforme, Funaria hygrometrica and Physcomitrella patens (Barthelmes 1940; Oehlkers and Bopp 1957; Engel 1968) and culminated in the synopses

Recent developments and perspectives of industrial rapeseed breeding

Due to substantial progress in breeding and cultivation practice rapeseed has become the world's third most important source of vegetable oil. Modification of the fatty acid composition to make rapeseed oil more competitive in various segments of the food and industrial oil markets has been an important objective of plant breeding and molecular genetics in recent years. While making up the primary demand by food and animal feed industry furnished by “double-low” quality rapeseed, so-called “canola”, interest increased to produce “Biodiesel” feedstocks or special materials being directed to several industrial niche markets, because of their higher value than commodity oils. Rapeseed oil is unique in having a large spectrum of usability and good properties for non-food applications, such as relatively homogeneous composition, high degree of refinement, freedom from contaminants, and also biodegradability, giving it advantages over petrochemicals. Consequently, one of the most important objectives of rapeseed breeding is the genetic modification of the seed oil by maximizing the proportion of specific fatty acids, like laurate, erucate or functionalized acids, in order to obtain tailor-made raw materials suited for industrial purposes.