Low external pH replaces 2,4-D in maintaining and multiplying 2,4-D-initiated embryogenic cells of carrot

A mixed culture comprised of both embryonic globules and nonembryogenic callus. was derived from seedling hypocotyls of Daucus carota cv. Scarlet Nantes on 2,4-D-containing medium using well-established methods. Then the mixed cultures were transferred to, and serially subcultured on, a hormone-free medium near pH 4. The medium contained 1 m M NH+ as the sole nitrogen source. When cultured in this way, embryonic globules were able to multiply without development into later embryo stages Nonembryogenic callus did not survive. Continuous culture of embryonic globules on this low pH hormone-free medium yielded cultures consisting entirely of preglobular stage proembryos (PGSPs). PGSP cultures have been maintained as such with continuous multiplication for nearly 2 years without loss of embryogenic potential. These hormone-free-maintained PGSPs continue their development to later embryo stages when cultured on the same hormone-free medium buffered at pH 5.8. We show that hormone-free medium near pH 4 can replace 2.4-D in its ability to sustain multiplication of 2,4-D-initiated embryogenic cells of carrot at an acceptable growth rate without their development into later embryo stages. This procedure provides selective conditions that do not permit the growth of nonembryogenic cells while providing an adequate environment for embryogenic cell proliferation and should prove invaluable in studying habituation.     

Somatic embryo development in carrot is associated with an increase in levels of S-adenosylmethionine,S-adenosylhomocysteine and DNA methylation

Almost homogeneous populations representing different developmental stages of somatic embryos (globular, torpedo-shaped, plantlets) and vacuolated cells were obtained from a cell suspension culture of carrot. The concentrations of S-adenosylmethionine (SAM), S-adenosylhomocysteine (SAH) and methylated DNA were determined in embryos at different developmental stages and were found to increase during somatic embryogenesis. The highest increase during embryogenesis was a 5-fold increase in the level of SAM. A considerable increase in the methylation index (SAM/SAH ratio) was also found. We propose that the levels of SAM and SAH may be involved in the control of somatic embryogenesis by affecting the level of DNA methylation, which in turn might cause differential changes in gene activation. An increase in the level of SAM may be a prerequisite for progression of embryogenesis and the development of complete embryos.     

An extracellular β-galactosidase secreted from cell suspension cultures of carrot. Its purification and involvement in cell wall-polysaccharide hydrolysis

β-Galactosidase (β-Galase, EC 3.2.1.23) activity has been detected in a culture medium of cell suspension cultures of carrot ( Daucus carota L. cv. Kintoki). The extracellular β-Galase (β-Galase-II) was purified to electrophoretic homogeneity from the concentrated medium using ammonium sulfate precipitation, chromatography on CM-Sephadex C-50. DEAE-Sepharose CL-6B and Sephacryl S-200HR, and preparative PAGE. The molecular mass of the purified enzyme was estimated to be 65 kDa by Sephacryl S-200HR gel-permeation, and 60 kDa by SDS-PAGE after treatment with SDS and 2-mercaptoethanol. The pI was 6.5. The K_m and V_max values for p -nitrophenyl (PNP)-β-D-galactopyranoside were 0.17 m M and 31.9 μmol (mg protein)^-1, h^-1, respectively. The optimal activity in McIlvaine's buffer occurred at pH 4.0–4.4. The enzyme activity was inhibited by Co²⁴, Cu²⁺, Hg^2-, p -chloromercuribenzoate (PCMB) and D-galactono-1,4-lactone. The enzyme acted on citrus galactan and larchwood arabinogalactan in an exo-fashion, and was slightly involved in the hydrolysis of an acidic pectic polymer containing arabinosyl and galactosyl residues and in the breakdown of cell walls isolated from carrot cell cultures.     

Intrinsic changes in photosynthetic parameters of carrot leaves under increasing CO<Subscript>2</Subscript> concentrations and soil moisture regimes

A controlled growth chamber experiment was conducted to investigate the short-term water use and photosynthetic responses of 30-d-old carrot seedlings to the combined effects of CO₂ concentration (50–1 050 μmol mol⁻¹) and moisture deficits (−5, −30, −55, and −70 kPa). The photosynthetic response data was fitted to a non-rectangular hyperbola model. The estimated parameters were compared for effects of moisture deficit and elevated CO₂ concentration (EC). The carboxylation efficiency (α) increased in response to mild moisture stress (−30 kPa) under EC when compared to the unstressed control. However, moderate (−55 kPa) and extreme (−70 kPa) moisture deficits reduced α under EC. Maximum net photosynthetic rate (P _Nmax) did not differ between mild water deficit and unstressed controls under EC. Moderate and extreme moisture deficits reduced P _Nmax by nearly 85 % compared to controls. Dark respiration rate (R _D) showed no consistent response to moisture deficit. The CO₂ compensation concentration (Γ) was 324 μmol mol⁻¹ for −75 kPa and ranged 63–93 μmol mol⁻¹ for other moisture regimes. Interaction between moisture deficit and EC was noticed for P _N, ratio of intercellular and ambient CO₂ concentration (C _i/C _a), stomatal conductance (g _s ), and transpiration rate (E). P _N was maximum and C _i/C _a was minimum at −30 kPa moisture deficit and at C _a of 350 μmol mol⁻¹. The g _s and E showed an inverse relationship at all moisture deficit regimes and EC. Water use efficiency (WUE) increased with moisture deficit up to −55 kPa and declined thereafter. EC showed a positive influence towards sustaining P _N and increasing WUE only under mild moisture stress, and no beneficial effects of EC were noticed at moderate or extreme moisture deficits.     

Immunolocalization of extensin and potato tuber lectin in carrot, tomato and potato

Tissue-specific expression of two members of the cell wall hydroxyproline-rich glycoprotein (HRGP) family, extensin and potato tuber lectin, was examined by immunolocalization at the light microscope level in various organs (leaves, stems, roots, fruit, tuber) of carrot ( Daucus carota cv. Thumbelina), tomato ( Lycopersicon esclentum cv. Pixie Hybrid II), and potato ( Solanum tuberosum cv. Kennebec). Extensin was prominently expressed in vascular tissue, particularly xylem and also phloem, although virtually all cells displayed some degree of staining which varied as a function of the tissue, organ, and plant under study. Antibodies against potato tuber lectin (PTL) displayed a localization pattern similar to that observed for extensin; notably PTL did not stain cambium but did stain epithelial cells lining secretory cavities. These distribution patterns are consistent with a role for extensin, and possibly PTL, in providing mechanical support in tissues subjected to compression or torsional stress imparted by vascular growth, or by similar stress brought about by transport of vascular fluids.     

Effects of water stress on plant volatile emission and insect oviposition preference in an agroecosystem

1. Drought has become more common and severe in many parts of the world due to climate change. The effect of water stress on insect oviposition preference that is key determinant for their fitness has received less attention.
2. Here, we examined how water stress may affect oviposition selection of Ostrinia furnacalis for maize plants in the greenhouse, and tested difference in volatile compounds emitted from treated maize, and electronantennogram and bioassay responses of O. furnacalis to the volatile profiles in maize plants.
3. Ostrinia furnacalis were more prone to lay eggs on the well-watered maize. Most plant volatile compounds differed significantly among the three water treatments, including increased emissions of β-caryophyllene, (E)-2-hexenal, and linalool, and decreased emission of (Z)-3-hexen-1-ol when subjected to increasing intensity of drought.
4. Varied volatile profiles of maize may drive oviposition decision of O. furnacalis, because O. furnacalis showed a clear oviposition preference for (Z)-3-hexen-1-ol, while not for β-caryophyllene, (E)-2-hexenal, and linalool, at the concentration of 1000 ppm.
5. This study advances understanding of drought effects on plant–insect interactions through volatile profiles. Our finding calls for attention to oviposition selection for insect pest management in agricultural settings, especially in regions under changing precipitation patterns.

     

Inheritance and plasticity of adult host acceptance in Yponomeuta species: implications for host shifts in specialist herbivores

Changes in host acceptance is an important factor in the host specialization of phytophagous insects, and knowledge of the genetic organization of this behaviour is necessary in order to understand how host shifts occur. Here we describe the inheritance of adult host acceptance (oviposition) in three closely related species of Yponomeuta Latreille (Lepidoptera: Yponomeutidae), and their interspecific hybrids. Yponomeuta cagnagellus (Hübner), a specialist on Euonymus europaeus L. (Celastraceae), Y. malinellus Zeller, a specialist on Malus spp. (Rosaceae), and Y. padellus (L.), oligophagous on a number of Rosaceae, were tested for their acceptance of parental hosts in choice tests. Acceptance of E. europaeus is semi‐dominant in hybrids of Y. cagnagellus×Y. padellus, and in hybrids of Y. cagnagellus×Y. malinellus. The dominance of this acceptance was confirmed in oviposition tests with backcross hybrids: backcross hybrids F1 × Y. cagnagellus oviposited mainly on E. europaeus and F1 × Y. padellus still deposited more than half of their egg masses on E. europaeus. Reciprocal hybrids did not differ in their host acceptance, indicating that the trait is autosomal. We further studied the effect of larval food on adult host acceptance (‘Hopkins host selection principle’) in split full‐sib F1 families. Larval diet influenced oviposition only in one of two hybrid crosses. The F1 hybrid of Y. padellus× Y. cagnagellus, reared on Prunus spinosa L., deposited a significantly lower percentage of egg masses on E. europaeus compared to their full‐sib sisters fed with E. europaeus. We did not find this in the reciprocal cross. However, still more than half of the egg masses are deposited on E. europaeus by hybrids that have no experience on this host. We conclude that the semi‐dominant character of acceptance of E. europaeus and a tendency of Rosaceae‐feeding Yponomeuta to deposit egg masses on this host may have created the opportunity for the host shift of the predecessor of Y. cagnagellus from Rosaceae to the Celastraceae. This shift may have been further facilitated by a weak tendency of adults to oviposit on their larval food source.     

Jack of all trades masters novel host plants: positive genetic correlations in specialist and generalist insect herbivores expanding their diets to novel hosts

One explanation for the widespread host specialization of insect herbivores is the 'Jack of all trades-master of none' principle, which states that genotypes with high performance on one host will perform poorly on other hosts. This principle predicts that cross-host correlation in performance of genotypes will be negative. In this study, we experimentally explored cross-host correlations and performance among families in four species (two generalist and two specialist) of leaf beetles (Cephaloleia spp.) that are currently expanding their diets from native to exotic plants. All four species displayed similar responses in body size, developmental rates and mortality rates to experimentally controlled diets. When raised on novel hosts, body size of larvae, pupae and adults were reduced. Development times were longer, and larval mortality was higher on novel hosts. Genotype × host-plant interactions were not detected for most traits. All significant cross-host correlations were positive. These results indicate very different ecological and evolutionary dynamics than those predicted by the 'Jack of all trades-master of none' principle.     

Nutrient composition of soil and plants may predict the distribution and abundance of specialist insect herbivores: implications for agent selection in weed biological control

Abstract  Refining the process of selecting specialist herbivores that are used as biological control agents to maximise effects on the targets can diminish the risk to non-target species. While biotic factors (e.g. plant demography and agent–host interactions) have been explored for clues to improve the way we make agent selection decisions, recent ecological research indicates that abiotic factors related to the habitat (e.g. plant and soil nutrient composition, and soil characteristics) are important predictors of insect herbivore community composition. In this paper we explore the relevance of plant nutrient composition to aid in selecting agents for the invasive Eurasian perennial Euphorbia esula (leafy spurge) in Illinois, USA. We propose that an approach that compares such abiotic factors across the native and invaded ranges of plants in conjunction with the community composition of specialist herbivores in the native range may yield valuable clues in selecting agents that are most likely to establish and regulate populations of the target weed.