Differential chemical allocation and plant adaptation: A Py-MS Study of 24 species differing in relative growth rate

The chemical composition of leaves of 24 wild species differing in potential relative growth rate (RGR) was analysed by pyrolysis-mass spectrometry. The variation in RGR significantly correlated with differences in chemical composition: slow-growing species were richer in glucan-based polysaccharides and in C16:0 fatty acid, whereas fast growing ones contained more protein (other than those incorporated in cell walls) and chlorophyll, sterols and diglycerides. Other, apparently significant correlations, e.g. for pentose-based hemicellulose and for guaiacyl lignin appeared solely based on a group separation between mono- and dicotyledonous species.Considering the eleven monocotyledonous and thirteen dicotyledonous species separately, correlations were found in addition to the previously mentioned general ones. Within the group of the monocotyledons the low-RGR species were significantly enriched in pentose-based hemicellulose, ferulic acid and (hydroxy)proline-rich cell wall protein and nearly significant in guaiacyl and syringyl lignin, fast-growing species contained more potassium. Within the group of the dicotyledons slow-growing species were enriched in triterpenes and aliphatic wax esters.In general, the monocotyledons contained more cell wall material such as pentose-based hemicellulose, ferulic acid, glucans (including cellulose) and guaiacyl-lignin, and also more aliphatic wax esters, than the dicotyledons. The dicotyledons, on the other hand, contained somewhat more protein than the grasses.Per unit weight of cell wall, the amount of (hydroxy)proline- rich protein in low-RGR species was comparatively low. A higher investment of cell wall proteins to explain the low rate of photosynthesis per unit of leaf nitrogen of slow-growing species as suggested by Lambers and Poorter (1992), therefore, seems unlikely.Abbreviations HPRP (hydroxy)proline-rich protein(s) - LAR leaf area ratio - LWR leaf weight ratio - MVA multivariate analysis - NAR net assimilation rate - PC principal component - PNUE photosynthetic nitrogen use efficiency - PyGCMS pyrolysis-gas chromatography-mass spectrometry - PyMS pyrolysis mass spectrometry - RGR relative growth rate - SLA specific leaf area - SLM specific leaf mass     

β‐1,3‐Glucanases in wheat and resistance to the Russian wheat aphid

The Russian wheat aphid (RWA), Diuraphis noxia (Mordvilko) is one of the most destructive insect pests of cereals world-wide. Although resistant cultivars have been bred, the biochemical mechanism of resistance is unknown. The aim of this work was to gain information on the mechanism of resistance which could contribute to more directed breeding of resistant cultivars in the future. The effect of RWA infestation on the inter- and intracellular β-1,3-glucanase activities was studied in different resistant wheat (Triticum aestivum L.) cultivars containing the Dn-1 gene for RWA resistance and corresponding near-isogenic susceptible cultivars. The activity was determined spectrophotometrically by measuring the release of glucose from laminarin. Infestation differentially induced the intra- and intercellular activities to much higher levels in resistant than susceptible cultivars within 48 h. According to immunological studies induced enzyme activities were due to increased protein levels. The intracellular β-1,3-glucanase contained about 8% exo-activity. The exo-activity made an insignificant contribution to the intercellular activity. The genetic background into which the resistance gene was bred did affect the level of activity that corresponded to the resistance performance. Seven apoplastic isoforms of β-1,3-glucanase, varying from acidic to basic, were resolved by isoelectric focusing. All isoenzymes were equally induced and no specific one could be linked to resistance or susceptibility. The RWA induced β-1,3-glucanase activity in resistant cultivars closely resembles defence responses during pathogenesis and seems to be part of a general defence response like the hypersensitive reaction (HR), which confers resistance to the RWA. This knowledge might be helpful in future to identify genes for RWA resistance. The increased β-1,3-glucanase activity after RWA infestation might serve as an additional measure to biochemically trace resistance in crosses during breeding.     

Chemical composition of 24 wild species differing in relative growth rate

The chemical composition of 24 plant species which showed a three-fold range in potential growth rate was investigated. The carbon content of whole plants was lower for fast-growing species than for slow-growing ones. Fast-growing species accumulated more organic N-compounds, organic acids and minerals, whereas slow-growing species accumulated more (hemi)cellulose, insoluble sugars and lignin. No correlations with relative growth rate were found for soluble phenolics, soluble sugars and lipids. The costs to construct 1 g of plant biomass were rather similar for fast- and slow-growing species, both when expressed as C needed for C-skeletons, as glucose to provide ATP and NAD(P)H, and as total glucose costs. Therefore, we conclude that, despite the differences in chemical composition between fast- and slow-growing species, variation in the costs of synthesis of whole plant biomass cannot explain interspecific variation in relative growth rate of herbaceous species.     

Differences in construction costs and chemical composition between deciduous and evergreen woody species are small as compared to differences among families

We tested to what extent differences in construction costs (CC) and chemical composition of woody species are attributed to leaf habit. Eight evergreen and eight deciduous species belonging to six families were selected to form eight phylogenetic independent contrasts (PICs). The plants were grown from seed in a glasshouse. Differences in leaf, stem and root CC between evergreen and deciduous species were minor, the proportion of variance explained by leaf habit generally being less than 6%. Surprisingly, differences in leaf chemical composition between deciduous and evergreen species were small as well. Variation in CC and chemical composition among families was substantial, the factor 'family' explaining 50-85% of variance. We therefore conclude that in this case, phylogeny is a more important factor than functional group. Leaves of the fast-growing species in this experiment showed high levels of minerals, organic acids, proteins and lipids, whereas leaves of the slow-growing species had higher concentrations of soluble phenolics, lignin as well as higher carbon/nitrogen (C/N) ratio. These relationships suggest a trade-off between growth and defence. In contrast, CC of leaves, stems, roots or whole plants showed no or only a weak correlation with relative growth rate (RGR). The C/N ratio of the leaves is an easily measured parameter that correlated strongly in a negative way with the RGR of the plants and reflected better the balance between investment in structure and physiological functioning than CC.     

Physiology and costs of resistance to herbivory and disease in Brassica

We used artificial selection experiments to study genetic allocation costs and physiological mechanisms of resistance to herbivory and fungal disease. Genetic costs to resistance were present in some instances and absent in others. Genetic resistance to the fungal pathogen, Leptosphaeria maculans was cost-free, while resistance to Peronospora parasitica showed a negative genetic correlation between disease resistance and growth rate. Leptosphaeria resistant genotypes had 13% higher chitinase activity. Genetic increases in myrosinase activity were correlated with increased resistance to flea beetles (Phyllotreta cruciferae), but resulted in lower plant fecundity, presumably due to production costs of myrosinase. Genetic costs of resistance may maintain genetic variation in natural plant populations. These studies demonstrate the predictive and explanatory power of a functional approach to plant-herbivore and plant-pathogen interactions.     

For or against: the importance of variation in growth rate for testing the EICA hypothesis

The evolution of increased competitive ability (EICA) hypothesis proposes that invasive species evolve decreased defense and increased competitive ability following natural enemy release. Previous studies have found evidence both for and against EICA. The resource-enemy release hypothesis (R-ERH) suggests that fast-growing species may experience stronger enemy release than slow-growing species. On the basis of R-ERH, the prediction of EICA will be held true for slow-growing genotypes, i.e., the slow-growing genotypes from the introduced range will be less resistant to herbivory and grow faster than those from the home range; while the EICA will not be held for fast-growing genotypes, i.e., there will be no significant differences in growth and defense traits between the introduced and native fast-growing genotypes. We tested these predictions preliminarily using five populations of the invasive plant Alternanthera philoxeroides. This species has two varieties in its home range, which showed a distinct growth-defense strategy: the northern A. p. var. acutifolia (Apa) had higher growth rate but lower resistance, while the southern A. p. var. obtusifolia (Apo) had lower growth rate but higher resistance level. Our results suggest that the EICA hypothesis is consistent with the slow-growing Apo, but not with the fast-growing Apa. We suggest that evolutionary changes in growth or resistance following enemy release are influenced by variation in growth rate within an invasive alien plant. These findings have important implications for the EICA hypothesis, and may partially explain why previous studies have found evidence both for and against EICA.     

A polysaccharide deacetylase from Puccinia striiformis f. sp. tritici is an important pathogenicity gene that suppresses plant immunity

The cell wall of filamentous fungi, comprised of chitin, polysaccharide and glycoproteins, maintains the integrity of hyphae and protect them from defence responses by potential host plants. Here, we report that one polysaccharide deacetylase of Puccinia striiformis f. sp. tritici (Pst), Pst_13661, suppresses Bax‐induced cell death in plants and Pst_13661 is highly induced during early stages of the interaction between wheat and Pst. Importantly, the transgenic wheat expressing the RNA interference (RNAi) construct of Pst_13661 exhibits high resistance to major Pst epidemic races CYR31, CYR32 and CYR33 by inhibiting growth and development of Pst, indicating that Pst_13661 is an available pathogenicity factor and is a potential target for generating broad‐spectrum resistance breeding material of wheat. It forms a homo‐polymer and has high affinity for chitin and germ tubes of Pst compared with the control. Besides, Pst_13661 suppresses chitin‐induced plant defence in plants. Hence, we infer that Pst_13661 may modify the fungal cell wall to prevent recognition by apoplastic surveillance systems in plants. This study opens new approaches for developing durable disease‐resistant germplasm by disturbing the growth and development of fungi and develops novel strategies to control crop diseases.     

The influence of genetics,defensive chemistry and the fungal microbiome on disease outcome in whitebark pine trees

The invasive fungal pathogen Cronartium ribicola infects and kills whitebark pine (Pinus albicaulis) throughout western North America. Whitebark pine has been proposed for listing under the Endangered Species Act in the USA, and the loss of this species is predicted to have severe impacts on ecosystem composition and function in high‐elevation forests. Numerous fungal endophytes live inside whitebark pine tissues and may influence the severity of C. ribicola infection, either directly by inhibition of pathogen growth or indirectly by the induction of chemical defensive pathways in the tree. Terpenes, a form of chemical defence in pine trees, can also influence disease. In this study, we characterized fungal endophyte communities in whitebark pine seedlings before and after experimental inoculation with C. ribicola, monitored disease progression and compared fungal community composition in susceptible vs. resistant seedlings in a common garden. We analysed the terpene composition of these same seedlings. Seed family identity or maternal genetics influenced both terpenes and endophyte communities. Terpene and endophyte composition correlated with disease severity, and terpene concentrations differed in resistant vs. susceptible seedlings. These results suggest that the resistance to C. ribicola observed in natural whitebark pine populations is caused by the combined effects of genetics, endophytes and terpenes within needle tissue, in which initial interactions between microbes and hosts take place. Tree genotype, terpene and microbiome combinations associated with healthy trees could help to predict or reduce disease severity and improve outcomes of future tree breeding programmes.     

The effect of riboflavin on the bacterial synthesis of vitamin B12-active material in the rat

On B₁₂-deficient rations that contained levels of the other B vitamins which were much higher than those required for optimal growth, a large proportion of negative control rats grew at a rate nearly equal to that of their B₁₂-fed littermates, while the others grew much more slowly. The average growth rate and the proportion of such fast-growing negative controls increased, in general, with increased concentrations of B vitamins in the ration.The feeding, to slow-growing negative controls on a high B vitamin diet, of a single small dose of feces from fast-growing negative controls on the same diet, brought about a markedly increased growth rate; inclusion of sulfasuxidine in the ration delayed this effect for several weeks in practically all cases. Feeding these feces to animals on moderate levels of the B vitamins failed to bring about increased growth. These results indicated bacterial synthesis of B₁₂-active material in the intestinal tract.Such synthesis was shown to be due to the high levels of dietary riboflavin. This finding is briefly discussed with regard to recent investigations on the chemical structure of vitamin B₁₂ and on related biochemical studies.     

Antibiotic resistance in potato cultivars to the aphid Myzus persicae

Commercially-grown potato cultivars were assessed for antibiotic resistance to Myzus persicae in a culture room at 19°C with a 2°C range and a 16 h photoperiod. The resistant rankings obtained were consistent for the following aphid metrics: maturation time, teneral adult weight, embryo complement and the production of nymphs. Intrinsic rates of increase (r_m) correlated with the above parameters of aphid growth and reproduction but this was not always the case for nymphal survival rate. The cultivars Ulster Tarn, Record and Maris Piper were relatively resistant to M. persicae and cultivars Desirée and King Edward were the most susceptible. When the aphid clone or the culture plants were changed there was no significant effect on the relative resistance of the potato cultivars under test. The relationship between the number of embryos in teneral adults and the production of nymphs by aphids was examined by grouping aphids in weight batches. There was a strong relationship between these two variables.     

Long-term impacts of elevated carbon dioxide and transgenic Bt cotton on performance and feeding of three generations of cotton bollworm

A field screening trial undertaken in Uganda showed that a number of Musa L. (Musaceae) cultivars and hybrids displayed high levels of resistance to banana weevil, Cosmopolites sordidus (Germar) (Coleoptera: Curculionidae), while most highland banana cultivars tended to be susceptible to weevil attack. In this study, research was undertaken to determine modalities of resistance to banana weevil. Laboratory studies suggested that all cultivars were attractive to the weevil and that females freely oviposited on all cultivars. While some differences were found between cultivars in attractivity and egg numbers, these were not related to subsequent damage. Therefore, antixenosis does not appear to play an important role in host‐plant resistance to banana weevil. Larval survivorship rates in living corms were, for the most part, low on resistant cultivars, suggesting that antibiosis mechanisms offer the primary avenues of resistance. In the laboratory, development was slower on some resistant cultivars although survivorship rates on excised corm material were not as well related to levels of resistance as that on living material. Sap appeared to play a minor role in reducing egg eclosion rates on some resistant cultivars. Methanol extracts from Kayinja, a resistant cultivar, inhibited larval development on corms of susceptible cultivars in the laboratory.     

Biochemical and bioassay analysis of resistance of potato (Solanum tuberosum L.) cultivars to attack by the slug Deroceras reticulatum (Muller)

A method is described for using young field slugs Deroceras reticulatum (Muller) in a bioassay study of biochemical resistance of potato (Solanum tuberosum L.) cultivars to slugs. Tuber parts or an artificial diet were provided as food sources. Comparisons were made of feeding, survival and weight gain between the susceptible cultivar Maris Piper and the resistant cultivar Pentland Dell. Biochemical analyses were made of these two cultivars and the resistant cultivars Stormont Enterprise and Majestic. Comparisons of tuber sections and peelings as food sources indicated factors affecting growth were located in the surface layers of the tubers. Phenolics and glycoalkaloids were concentrated in the surface layers but the amounts were similar in the susceptible and resistant cultivars and the bioassays indicated that neither acting alone could explain resistance. The amounts and distribution of free amino acids also did not correlate with resistance although when supplied in the artificial diet they partly inhibited feeding. Proteinaceous inhibitors of slug gut proteolytic enzymes were present throughout the tubers but were not concentrated in the surface layers and the amounts were similar in the different cultivars thus they too did not explain the difference in susceptibility between the cultivars. Bioassays using acetone extracts (low molecular weight substances) and acetone powders (high molecular weight substances) either alone or in combination indicated that the resistant cultivar Pentland Dell contained a high molecular weight substance which together with a low molecular weight substance from either the same cultivar or the susceptible Maris Piper could confer resistance. Bioassays using protein extracts supplied in the presence or absence of chlorogenic acid indicated that this mechanism could comprise enzymic oxidation of phenolics. Assays of phenolase confirmed this since activity was highest in the outer layers of the tubers and was highest in the three resistant cultivars. Thus the chief resistance factor identified was high phenolase activity acting rapidly on phenolics when the slug first bites the tuber surface. The quantity of phenolics per se did not control the resistance. Thus while phenolics must be available, resistance is compatible with low blackening on cutting the tuber.     

Feeding‐induced changes in defence enzymes and PR proteins and their implications in host resistance to Nilaparvata lugens

Six rice genotypes showing susceptible and resistant reactions to brown planthopper (BPH), Nilaparvata lugens were studied for feeding‐induced changes in defence enzymes and pathogenesis‐related (PR) proteins. The high resistant genotypes PTB 33, ADT 45 and ASD 7 and moderately resistant genotypes CO 43 and KAU 1661 recorded the greater expression of defence enzymes peroxidase, polyphenol oxidase, phenylalanine ammonia lyase, total phenol and β‐1,3 glucanase in response to N. lugens feeding at 1 day after infestation (DAI) compared with susceptible genotype TN1. The greater activity of chitinase was observed in resistant cultivars at 3 DAI and the activity was sustained for more than 1 week compared with susceptible TN1. In conclusion, the current study revealed that these defence enzymes and PR proteins might attribute to the resistance mechanisms in rice plants against BPH infestation.     

Defence response of pepper (Capsicum annuum) suspension cells to Phytophthora capsici

Cell suspension cultures of three cultivars of Capsicum annuum L., with different degrees of sensibility to the fungus Phytophthora capsici, responded to elicitation by both lyophilized mycelium and fungus filtrate. They showed conductivity changes, browning, production of the phytoalexin capsidiol and synthesis or accumulation of pathogenesis-related (PR) proteins with glucanase (EC 3.2.1.39) and chitinase (EC 3.2.1.14) activities. The cultivation medium was optimised for growth of both the plant and the fungus in order to avoid any stress during their combination. The resistant cv. Smith-5, showed a more rapid and intense response to the elicitor preparations than the sensitive cvs Americano and Yolo Wonder. This was particularly evident when the cell suspensions were elicited with the filtrate, when differences became clearly visible after only 6 h incubation. The greatest rate of capsidiol accumulation occurred after 18 h in the mycelium-elicited cells and after 12 h in those elicited with the filtrate. These times are the optimal for capsidiol accumulation, and the phytoalexin is produced much more rapidly than it can be excreted into the extracellular medium. The inhibition threshold of fungal growth (300 µg capsidiol [g dry weight]^?1) was reached only in the resistant cultivar. The induction of an intracellular glucanase (pI 8.9 and R_f 0.18) and an extracellular chitinase (pI 5.4 and R_f 0.70) only in the resistant cultivar 24 h after elicitation suggests that these enzymes are involved in the resistance to Phytophthora capsici, while other hydrolases common to all three cultivars form part of a more general defence. The results indicate that elicitation of pepper cell suspension cultures by signal molecules from P. capsici exhibits properties of a multicomponent dynamic system in which different protective mechanisms play complementary roles in the overall expression of the defence reaction. We confirm that the differential responses of resistant and susceptible pepper cultivars to P. capsici previously seen in plant stem sections are retained in suspension culture.     

Molecular structure of the resistant biopolymer in zygospore cell walls of Chlamydomonas monoica

The unicellular green alga Chlamydomonas monoica Strehlow is known to produce zygospores with a cell wall that is resistant against microbial and chemical attack. This resistance is thought to be due to the presence of a sporopollenin-like material. However, the resistant nature of sporopollenin-like materials seriously hampers their structural analysis. With complementary techniques such as ¹³C-nuclear magnetic resonance spectroscopy, Curie-point pyrolysis-gas chromatography/mass spectroscopy and RuO₄ chemical degradation, the chemical composition of resistant biopolymer in the isolated cell walls of C. monoica zygospores was determined. This material is composed of C₂₂–C₃₀ linear alcohols and carboxylic acids, intermolecularly linked via ester and ether-linkages similar to the resistant aliphatic biopolymers encountered in the walls of the vegetative cells of the algae Tetraedron minimum, Scenedesmus communis and Pediastrum boryanum. Received: 29 April 1998 / Accepted: 2 October 1998     

Ultrastructural and Immunocytochemical Studies on Effects of Barley Yellow Dwarf Virus – Infection on Fusarium Head Blight, Caused by Fusarium graminearum, in Wheat Plants

The interactions between barley yellow dwarf virus (BYDV) and Fusarium head blight (FHB), caused by Fusarium graminearum, were studied in the two winter wheat cultivars (cvs.), Agent (susceptible to FHB) and Petrus (moderately resistant to FHB), using ultrastructural and immunocytochemical methods. Infections of wheat plants of both cvs. by BYDV increased susceptibility to FHB. BYDV infection caused numerous cytological changes in lemma tissue of both cvs. such as formation of vesicles in the cytoplasm, degradation of fine structures of chloroplasts of both cvs. and accumulation of large starch grains in the chloroplasts. Electron microscopical studies showed that the development of F. graminearum on spike surfaces was not affected in BYDV‐infected plants. After penetration and intercellular growth in lemma tissue, defence responses to Fusarium infections were markedly reduced in BYDV‐diseased plants compared to the tissue of virus‐free plants. At sites of contact of fungal cells with host tissue, depositions of cell wall material were distinctly less pronounced than in tissues of virus‐free plants of cv. Petrus. Detection of β‐1,3‐glucanases and chitinases in lemma tissue of cv. Agent revealed no appreciably increased accumulation of both defence enzymes in F. graminearum‐infected virus‐free and BYDV‐infected tissues compared to the non‐infected control tissue. On the other hand, in cv. Petrus, infection with F. graminearum induced a markedly enhanced activity of both enzymes 3 days after inoculation. The increase of both enzyme activities was less pronounced in BYDV‐infected plants than in tissue exclusively infected with F. graminearum. Cytological studies suggest that in contrast to the susceptible cv. Agent postinfectional defence responses may play still an important role in the resistance of the moderately resistant cv. Petrus to FHB.