Plant Protection Institute

Abstract The number of angiosperm species for which nuclear DNA amount estimates have been made has nearly trebled since the last collected lists of such values were published, and therefore, publication of a more comprehensive list is overdue. This paper lists absolute nuclear DNA amounts for 753 angiosperm species. The data were assembled primarily for reference purposes, and so the species are listed in alphabetical order, as this was felt to be more helpful to cyto- and biochemists whom, it is anticipated, will be among its major users. The paper also reviews aspects of the history, nomenclature, methods, accuracy and problems of nuclear DNA estimation in angiosperms. No attempt is made to reconsider those aspects of nuclear DNA estimation which have been fully revised previously, although the bibliography of such aspects is given. Instead, the paper is intended as a source of basic information regarding the terminology, practice and limitations of nuclear DNA estimation, especially by Feulgen microdensitometry, as currently practiced.

Disease resistance strategies are powerful approaches to sustainable agriculture because they reduce chemical input into the environment. Recently, Piriformospora indica, a plant-root-colonizing basidiomycete fungus, has been discovered in the Indian Thar desert and was shown to provide strong growth-promoting activity during its symbiosis with a broad spectrum of plants. Here, we report on the potential of P. indica to induce resistance to fungal diseases and tolerance to salt stress in the monocotyledonous plant barley. The beneficial effect on the defense status is detected in distal leaves, demonstrating a systemic induction of resistance by a root-endophytic fungus. The systemically altered "defense readiness" is associated with an elevated antioxidative capacity due to an activation of the glutathione-ascorbate cycle and results in an overall increase in grain yield. Because P. indica can be easily propagated in the absence of a host plant, we conclude that the fungus could be exploited to increase disease resistance and yield in crop plants.

Summary Experiments were carried out to assess the increase in yield potential of winter wheat in the U.K. due to variety improvement since the early years of this century. The effects of other genetic changes were minimized by applying fungicide to control eyespot and foliar diseases, and by using nets to prevent lodging. The experiments were carried out in 1978 at Cambridge. One, on soil of high fertility in Camp Field, received 104 kg N/ha and the other, on soil of lower fertility in Paternoster Field, received 38 kg N/ha. Twelve genotypes were tested. Eight were varieties which formed a chronological series beginning with Little Joss, introduced in 1908. The remaining genotypes were recently developed selections from the Plant Breeding Institute and a line bred by the French breeders, Benoist. The average yield of the 12 varieties and lines tested was 3·96 t/ha in Paternoster Field and 6·40 t/ha in Camp Field. In both fields the two highest yielding entries, Hobbit and the advanced breeding line 989/10, outyielded Little Joss by close to 40%. Benoist 10483 was the only entry for which the percentage yield advantage depended on high soil fertility. The newer, high yielding, varieties were shorter and reached anthesis earlier than the older varieties. They had lower stem weights/m 2 than the older varieties but similar maximum leaf area indices and leaf weights/m 2 . Within each experiment the total dry-matter production of the varieties was similar, the increase in grain yield due to variety improvement being associated mainly with greater harvest index (ratio of grain yield to grain + straw yield). It is argued that by a continuation of the trend towards reduced stem length, with no change in above-ground biomass, breeders may be able to increase harvest index, from the present value of about 50% to about 60%, achieving a genetic gain in yield of some 25%. As the limit to harvest index is approached, genetic gain in yield will depend on detecting and exploiting genetic variation in biomass production.

Legume plants host nitrogen-fixing endosymbiotic Rhizobium bacteria in root nodules. In Medicago truncatula, the bacteria undergo an irreversible (terminal) differentiation mediated by hitherto unidentified plant factors. We demonstrated that these factors are nodule-specific cysteine-rich (NCR) peptides that are targeted to the bacteria and enter the bacterial membrane and cytosol. Obstruction of NCR transport in the dnf1-1 signal peptidase mutant correlated with the absence of terminal bacterial differentiation. On the contrary, ectopic expression of NCRs in legumes devoid of NCRs or challenge of cultured rhizobia with peptides provoked symptoms of terminal differentiation. Because NCRs resemble antimicrobial peptides, our findings reveal a previously unknown innovation of the host plant, which adopts effectors of the innate immune system for symbiosis to manipulate the cell fate of endosymbiotic bacteria.

Fungi and plants are rich sources of thousands of secondary metabolites. The genetically coded possibilities for secondary metabolite production, the stimuli of the production, and the special phytotoxins basically determine the microscopic fungi-host plant interactions and the pathogenic lifestyle of fungi. The review introduces plant secondary metabolites usually with antifungal effect as well as the importance of signaling molecules in induced systemic resistance and systemic acquired resistance processes. The review also concerns the mimicking of plant effector molecules like auxins, gibberellins and abscisic acid by fungal secondary metabolites that modulate plant growth or even can subvert the plant defense responses such as programmed cell death to gain nutrients for fungal growth and colonization. It also looks through the special secondary metabolite production and host selective toxins of some significant fungal pathogens and the plant response in form of phytoalexin production. New results coming from genome and transcriptional analyses in context of selected fungal pathogens and their hosts are also discussed.

The root endophytic basidiomycete Piriformospora indica has been shown to increase resistance against biotic stress and tolerance to abiotic stress in many plants. Biochemical mechanisms underlying P. indica-mediated salt tolerance were studied in barley (Hordeum vulgare) with special focus on antioxidants. Physiological markers for salt stress, such as metabolic activity, fatty acid composition, lipid peroxidation, ascorbate concentration and activities of catalase, ascorbate peroxidase, dehydroascorbate reductase, monodehydroascorbate reductase and glutathione reductase enzymes were assessed. Root colonization by P. indica increased plant growth and attenuated the NaCl-induced lipid peroxidation, metabolic heat efflux and fatty acid desaturation in leaves of the salt-sensitive barley cultivar Ingrid. The endophyte significantly elevated the amount of ascorbic acid and increased the activities of antioxidant enzymes in barley roots under salt stress conditions. Likewise, a sustained up-regulation of the antioxidative system was demonstrated in NaCl-treated roots of the salt-tolerant barley cultivar California Mariout, irrespective of plant colonization by P. indica. These findings suggest that antioxidants might play a role in both inherited and endophyte-mediated plant tolerance to salinity.

Plant glutathione S-transferases (GSTs) are ubiquitous and multifunctional enzymes encoded by large gene families. A characteristic feature of GST genes is their high inducibility by a wide range of stress conditions including biotic stress. Early studies on the role of GSTs in plant biotic stress showed that certain GST genes are specifically up-regulated by microbial infections. Later numerous transcriptome-wide investigations proved that distinct groups of GSTs are markedly induced in the early phase of bacterial, fungal and viral infections. Proteomic investigations also confirmed the accumulation of multiple GST proteins in infected plants. Furthermore, functional studies revealed that overexpression or silencing of specific GSTs can markedly modify disease symptoms and also pathogen multiplication rates. However, very limited information is available about the exact metabolic functions of disease-induced GST isoenzymes and about their endogenous substrates. The already recognized roles of GSTs are the detoxification of toxic substances by their conjugation with glutathione, the attenuation of oxidative stress and the participation in hormone transport. Some GSTs display glutathione peroxidase activity and these GSTs can detoxify toxic lipid hydroperoxides that accumulate during infections. GSTs can also possess ligandin functions and participate in the intracellular transport of auxins. Notably, the expression of multiple GSTs is massively activated by salicylic acid and some GST enzymes were demonstrated to be receptor proteins of salicylic acid. Furthermore, induction of GST genes or elevated GST activities have often been observed in plants treated with beneficial microbes (bacteria and fungi) that induce a systemic resistance response (ISR) to subsequent pathogen infections. Further research is needed to reveal the exact metabolic functions of GST isoenzymes in infected plants and to understand their contribution to disease resistance.

DNA phylogenetic comparisons have shown that morphology-based species recognition often underestimates fungal diversity. Therefore, the need for accurate DNA sequence data, tied to both correct taxonomic names and clearly annotated specimen data, has never been greater. Furthermore, the growing number of molecular ecology and microbiome projects using high-throughput sequencing require fast and effective methods for en masse species assignments. In this article, we focus on selecting and re-annotating a set of marker reference sequences that represent each currently accepted order of Fungi. The particular focus is on sequences from the internal transcribed spacer region in the nuclear ribosomal cistron, derived from type specimens and/or ex-type cultures. Re-annotated and verified sequences were deposited in a curated public database at the National Center for Biotechnology Information (NCBI), namely the RefSeq Targeted Loci (RTL) database, and will be visible during routine sequence similarity searches with NR_prefixed accession numbers. A set of standards and protocols is proposed to improve the data quality of new sequences, and we suggest how type and other reference sequences can be used to improve identification of Fungi. Database URL: http://www.ncbi.nlm.nih.gov/bioproject/PRJNA177353.

Abstract The genes that code for endosperm storage proteins occur at nine complex loci on six different chromosomes. Glu-A1, Glu-B1 and Glu-D1 contain the genes for high molecular mass subunits of glutenin and are close to the centromere on the long arms of chromosomes 1A, 1B and 1D respectively. On the short arms of the same chromosomes, but distant from the centromere, are Gli-A1, Gli-B1 and Gli-D1. Each of these loci carry three major gene families coding for ω-gliadins, γ-gliadins and low molecular mass glutenin subunits. The remaining loci, Gli-A2, Gli-B2 and Gli-D2 occur near the ends of the short arms of chromosomes 6A, 6B and 6D respectively and each code for α- and β-gliadins. Recombination of genes within a locus is very rare and has so far been detected only at Glu-B1, at the rate of about one recombinant in 1000 progeny. Each locus displays allelic variation and this is responsible for differences among varieties in protein quality for making bread. The protein variants that are associated with good quality are being identified, firstly by analysing segregating populations and secondly from the development of near-isogenic lines. Current, incomplete, information on the relative qualities of different alleles at each locus indicates the following order of im portance: Glu-1 > Gli-1 > Gli-2. Landraces of primitive agriculture are being screened for novel proteins. The genes for some of them are being incorporated into the genomes of commercial wheats.

Abstract Nuclear DNA amounts have been estimated for more than 200 angiosperm species since the last collected list of such values for about 750 species was published by Bennett & Smith in 1976 (Phil. Trans. R. Soc. Lond. B 274, 227- 274). These new estimates are either scattered in a wide range of scientific journals or, in many cases, unpublished; so they are not readily accessible. A publication, collecting these data in a single list is required. This paper contains a supplementary list of absolute DNA values, including estimates for 240 angiosperm species not listed by Bennett & Smith in 1976, as well as additional estimates for 41 species already listed by them. These data are assembled primarily for reference purposes. Consequently, the species are listed in alphabetical order, as this was felt to be more helpful to cyto- and biochemists, who it is anticipated will be among the major users.

We analyzed the pathogenesis-related generation of H2O2 using the microscopic detection of 3,3-diaminobenzidine polymerization in near-isogenic barley (Hordeum vulgare L.) lines carrying different powdery mildew (Blumeria graminis f.sp. hordei) resistance genes, and in a line expressing chemically activated resistance after treatment with 2,6-dichloroisonicotinic acid (DCINA). Hypersensitive cell death in Mla12 and Mlg genotypes or after chemical activation by DCINA was associated with H2O2 accumulation throughout attacked cells. Formation of cell wall appositions (papillae) mediated in Mlg and mlo5 genotypes and in DCINA-activated plants was paralleled by H2O2 accumulation in effective papillae and in cytosolic vesicles of up to 2 μm in diameter near the papillae. H2O2 was not detected in ineffective papillae of cells that had been successfully penetrated by the fungus. These findings support the hypothesis that H2O2 may play a substantial role in plant defense against the powdery mildew fungus. We did not detect any accumulation of salicylic acid in primary leaves after inoculation of the different barley genotypes, indicating that these defense responses neither relied on nor provoked salicylic acid accumulation in barley.

Abstract A review of the literature showed that spider abundance was increased by diversification in 63% of studies. A comparison of diversification modes showed that spider abundance in the crop was increased in 33% of studies by ‘aggregated diversification’ (e.g. intercropping and non‐crop strips) and in 80% of studies by ‘interspersed diversification’ (e.g., undersowing, partial weediness, mulching and reduced tillage). It is suggested that spiders tend to remain in diversified patches and that extending the diversification throughout the whole crop (as in interspersed diversification) offers the best prospects for improving pest control. There is little evidence that spiders walk in significant numbers into fields from uncultivated field edges, but diversification at the landscape level serves to foster large multi‐species regional populations of spiders which are valuable as a source of aerial immigrants into newly planted crops. There are very few manipulative field studies where the impact of spiders on pests has been measured in diversified crops compared with undiversified controls. It is encouraging, however, that in those few studies an increased spider density resulted in improved pest control. Future work needs are identified.

Summary An analysis of incidence of Phytophthora spp. in 732 European nurseries producing forest transplants, larger specimen trees, landscape plants and ornamentals, plus 2525 areas in which trees and shrubs were planted, is presented based on work conducted by 38 research groups in 23 European countries between 1972 and 2013. Forty‐nine Phytophthora taxa were recorded in 670 nurseries (91.5%); within these nurseries, 1614 of 1992 nursery stands (81.0%) were infested, although most affected plants appeared healthy. In forest and landscape plantings, 56 Phytophthora taxa were recovered from 1667 of 2525 tested sites (66.0%). Affected plants frequently showed symptoms such as crown thinning, chlorosis and dieback caused by extensive fine root losses and/or collar rot. Many well‐known highly damaging host– Phytophthora combinations were frequently detected but 297 and 407 new Phytophthora –host associations were also observed in nurseries and plantings, respectively. On average, 1.3 Phytophthora species/taxa per infested nursery stand and planting site were isolated. At least 47 of the 68 Phytophthora species/taxa detected in nurseries and plantings were exotic species several of which are considered well established in both nurseries and plantings in Europe. Seven known Phytophthora species/taxa were found for the first time in Europe, while 10 taxa had not been previously recorded from nurseries or plantings; in addition, 5 taxa were first detections on woody plant species. Seven Phytophthora taxa were previously unknown to science. The reasons for these failures of plant biosecurity in Europe, implications for forest and semi‐natural ecosystems and possible ways to improve biosecurity are discussed.

Last instar larvae of Manduca sexta (Johanssen) and Heliothis zea (Boddie), fed on different host plants or on artificial diet, and then tested individually, have shown clear preference for the plant previously eaten. This induced preference is specific for the inducing plant species and is not merely a change in the insect's general threshold of food acceptability. The extent to which preference can be induced by various host plants differs considerably. No induction is possible with plants outside the insect's host range. In larvae fed on artificial diet preference to a given host plant can be induced even by a one‐day feeding on it. A preference once induced is not wiped out by two larval moults and subsequent feeding on an artificial diet. Thus it is supposed that the information serving as a basis for the induced feeding habit is stored in the central nervous system. The possible ecological significance of the induced preference is discussed. ZUSAMMENFASSUNG INDUZIERTE NAHRUNGSPRÄFERENZ BEI SCHMETTERLINGSRAUPEN Larven des letzten Entwicklungsstadium von Manduca sexta (Sphingidae) und Heliothis zea (Noctuidae), die an verschiedenen Wirtspflanzen gezüchtet wurden, bevorzugten in individuellen Nahrungswahlversuchen jene Pflanzenarten, an welchen sie sich entwickelten. Diese induzierte Präferenz ist spezifisch für die induzierende Pflanzenart und nicht einfach eine allgemeine Verschiebung der Reizschwelle für Frass‐ bzw. Vergällungsstoffe. Die Stärke der durch verschiedene Wirtspflanzen induzierten Präferenz variierte von Pflanze zu Pflanze erheblich. Es war nicht möglich, Präferenz für eine Pflanzenart ausserhalb des Wirtspflanzenkreises zu induzieren. In den an künstlicher Nahrung gezüchteten Raupen entwickelte sich die Präferenz für eine gegebene Wirtspflanze bereits während einer eintägigen Ernährung an derselben. Die einmal erworbene Präferenz wurde selbst durch zwei Häutungen und Ernährung mit künstlicher Nahrung nicht ausgelöscht. Demnach wird angenommen, dass die als Basis für die induzierte Präferenz dienende Rezeptoreninformation im Zentralnervensystem gespeichert wird. Die wahrscheinliche ökologische Bedeutung der induzierten Präferenz wird besprochen.

Heavy metal (HM) toxicity has become a global concern in recent years and is imposing a severe threat to the environment and human health. In the case of plants, a higher concentration of HMs, above a threshold, adversely affects cellular metabolism because of the generation of reactive oxygen species (ROS) which target the key biological molecules. Moreover, some of the HMs such as mercury and arsenic, among others, can directly alter the protein/enzyme activities by targeting their -SH group to further impede the cellular metabolism. Particularly, inhibition of photosynthesis has been reported under HM toxicity because HMs trigger the degradation of chlorophyll molecules by enhancing the chlorophyllase activity and by replacing the central Mg ion in the porphyrin ring which affects overall plant growth and yield. Consequently, plants utilize various strategies to mitigate the negative impact of HM toxicity by limiting the uptake of these HMs and their sequestration into the vacuoles with the help of various molecules including proteins such as phytochelatins, metallothionein, compatible solutes, and secondary metabolites. In this comprehensive review, we provided insights towards a wider aspect of HM toxicity, ranging from their negative impact on plant growth to the mechanisms employed by the plants to alleviate the HM toxicity and presented the molecular mechanism of HMs toxicity and sequestration in plants.

We present the DNA sequence and plant-tumor transcription pattern of some 2400 base pairs from the right border region of pTi T37 DNA from the virulent Agrobacterium tumefaciens strain T37. This region includes the entire transcription unit encompassing the nopaline synthase gene, together with parts of other transcription units. The strategy used to determine the sequence also produced two opposing series of defined, asymmetric deletions across the target DNA region, some of which may serve future purposes in the exploitation of this sequence, which is known to be expressed in a wide variety of host plant tissues.

Abstract Political efforts are made in the European Union (EU) to reduce pesticide use and to increase the implementation of integrated pest management (IPM). Within the EU project ENDURE, research priorities on pesticide reduction are defined. Using maize, one of the most important crops in Europe, as a case study, we identified the most serious weeds, arthropod pests, and fungal diseases as well as classes and amounts of pesticides applied. Data for 11 European maize growing regions were collected from databases, publications and expert estimates. Silage maize dominates in northern Europe and grain production in central and southern Europe. Crop rotations range from continuous growing of maize over several years to well‐planned rotation systems. Weeds, arthropod pests and fungal diseases cause economic losses in most regions, even though differences exist between northern countries and central and southern Europe. Several weed and arthropod species cause increasing problems, illustrating that the goal of reducing chemical pesticide applications is challenging. Pesticides could potentially be reduced by the choice of varieties including genetically modified hybrids, cultural control including crop rotation, biological control, optimized application techniques for chemicals, and the development of more specific treatments. However, restrictions in the availability of alternative pest control measures, farm organization, and the training and knowledge of farmers need to be overcome before the adoption of environmentally friendly pest control strategies can reduce chemical pesticides in an economically competitive way. The complex of several problems that need to be tackled simultaneously and the link between different control measures demonstrates the need for IPM approaches, where pest control is seen in the context of the cropping system and on a regional scale. Multicriteria assessments and decision support systems combined with pest monitoring programs may help to develop region‐specific and sustainable strategies that are harmonized within a EU framework.

Abstract A survey of work on meiotic duration in diploid plants shows that the duration is positively correlated with the DNA content per nucleus and with the mitotic cycle time. However, meiotic duration is not solely determined by the DNA content per nucleus but is also affected by chromosomal organization, DNA structure and the developmental pattern of the organism. Thus, in three polyploid plant species meiosis is much shorter and in three animal species it is much longer than would be expected in diploid plant species having corresponding DNA contents. Differences in meiotic duration in plant species are usually the result of proportional differences in all the stages of meiosis. Factors affecting the initiation, control and duration of meiosis are discussed. The consequences of changes in nuclear DNA content on developmental processes and the life cycle in plants are considered. It is suggested that DNA influences development in two ways, first directly through its informational content, and second indirectly by the physical mechanical effects of its mass independent of its informational content.

Most higher plant genomes contain a high proportion of repeated sequences. Thus repetitive DNA is a major contributor to plant chromosome structure. The variation in total DNA content between species is due mostly to variation in repeated DNA content. Some repeats of the same family are arranged in tandem arrays, at the sites of heterochromatin. Examples from the Secale genus are described. Arrays of the same sequence are often present at many chromosomal sites. Heterochromatin often contains arrays of several unrelated sequences. The evolution of such arrays in populations is discussed. Other repeats are dispersed at many locations in the chromosomes. Many are likely to be or have evolved from transposable elements. The structures of some plant transposable elements, in particular the sequences of the terminal inverted repeats, are described. Some elements in soybean, antirrhinum and maize have the same inverted terminal repeat sequences. Other elements of maize and wheat share terminal homology with elements from yeast, Drosophila, man and mouse. The evolution of transposable elements in plant populations is discussed. The amplification, deletion and transposition of different repeated DNA sequences and the spread of the mutations in populations produces a turnover of repetitive DNA during evolution. This turnover process and the molecular mechanisms involved are discussed and shown to be responsible for divergence of chromosome structure between species. Turnover of repeated genes also occurs. The molecular processes affecting repeats imply that the older a repetitive DNA family the more likely it is to exist in different forms and in many locations within a species. Examples to support this hypothesis are provided from the Secale genus.

Plant growth-promoting bacteria play an essential role in enhancing the physical, chemical and biological characters of soils by facilitating nutrient uptake and water flow, especially under abiotic stress conditions, which are major constrains to agricultural development and production. Drought is one of the most harmful abiotic stress and perhaps the most severe problem facing agricultural sustainability, leading to a severe shortage in crop productivity. Drought affects plant growth by causing hormonal and membrane stability perturbations, nutrient imbalance and physiological disorders. Furthermore, drought causes a remarkable decrease in leaf numbers, relative water content, sugar yield, root yield, chlorophyll a and b and ascorbic acid concentrations. However, the concentrations of total phenolic compounds, electrolyte leakage, lipid peroxidation, amounts of proline, and reactive oxygen species are considerably increased because of drought stress. This negative impact of drought can be eliminated by using plant growth-promoting bacteria (PGPB). Under drought conditions, application of PGPB can improve plant growth by adjusting hormonal balance, maintaining nutrient status and producing plant growth regulators. This role of PGPB positively affects physiological and biochemical characteristics, resulting in increased leaf numbers, sugar yield, relative water content, amounts of photosynthetic pigments and ascorbic acid. Conversely, lipid peroxidation, electrolyte leakage and amounts of proline, total phenolic compounds and reactive oxygen species are decreased under drought in the presence of PGPB. The current review gives an overview on the impact of drought on plants and the pivotal role of PGPB in mitigating the negative effects of drought by enhancing antioxidant defense systems and increasing plant growth and yield to improve sustainable agriculture.