Morphogenesis of insect-induced plant galls: facts and questions

Insect-induced galls (‘galls’ hereafter) represent highly regulated growth manifestations on plants. They present unique geometrical forms, which are, usually, unknown in the normal plant system. Galls are the best examples for modified natural structures that arise solely because of messages from an alien organism - the insect. Galls develop as an extension of the host-plant phenotype. But how the physiological networks and signal-activated subsystems work in coordination in expressing galls that serve the nutritional and shelter needs of the inducing insect are unclear. In galls and bacteria-induced tumors, the basic developmental events are essentially similar. However, tightly regulated specific differentiation processes occur in galls, making them different from tumors. Moreover variations in differentiation patterns occur in galls induced by insects of different taxonomic groups. While providing an overview of the control of shape and structure in galls, this article identifies the unanswered questions in gall morphogenesis.By analyzing the recognizable steps in gall morphogenesis, viz., gall initiation, stimulus recognition in host plants, signal transduction in host plants, growth of galls, and qualitative differentiation in galls, I have indicated that the insect saliva flushed on the wounded plant site alters the subcellular environment of cells and thus places it in a state of chemical shock. This shock induces osmotic changes, which establishes the first recognizable stage in gall induction. To repair the wound and neutralize the osmotic-change induced stress, the plant responds by establishing from one to a few metaplasied cell(s). Localized metabolic changes spread, from these cells, not throughout the involved plant organ, but in a limited manner around the immediate site of insect occurrence. When the shock is of low intensity, the plant responds with the development of one or more metaplasied cell(s) and gall development starts; when the shock factor is of high intensity, the cells under the insect action die, rejecting the inducing insect, defending plant tissue. These changes dictate the new morphogenetic events. Insects feed on gall tissue continuously for a specific period (synchronizing with their life history) and therefore, the osmotic-change related stress prevails for that span of time, which in turn triggers a sequence of plant-mediated changes including synthesis of growth promotors. Osmotic stress affects electrical properties of the plasma membrane and impacts on IAA activity, which in turn, alters H⁺-transport systems. During the physical action of insect feeding, the host-cell wall breaks down, and the degenerated wall materials act as elicitors.Using galls (e.g., ‘cecidial shoots’ on leaves, modified vegetative buds) induced on species of south and south-east Asian Dipterocarpaceae by different Beesoniidae (Coccoidea) as model complexity in gall morphogenesis is discussed. Manipulatory experimental studies done on the regeneration of epiphyllous buds on Pteridium, Begonia, and a Helianthus hybrid indicate that insect-induced neoplasmic shoots that arise on the leaves of tropical Dipterocarpaceae fall into the morphogenetic regulation of leaf, yet maintaining their freedom of differentiation. Even though a gall is a part of the plant - a multicellular organism made of the same genetic material - organismal development generates a range of cell types with dictated functions fitting into of Waddington's epigenetic-landscape model. As of today, our knowledge stops here.Plants as living systems display different strategies to mitigate and neutralize stress. Although these strategies exist in their genetic constitution, they are mediated by complex molecular interactions. Plants have a flexible short-term strategy to respond to stress; organisms that can modify gene expression reversibly have an advantage in evolutionary terms, since they can avoid rearrangements and species diversification. Mechanisms of DNA methylation and histone modifications possibly regulate inheritance of stress ‘memories’. Inherited genetic traits also play a role in gall morphogenesis, followed by roles played by correlating morphogenetic factors. An articulated reconstruction of the developmental process commencing from either one or a group of metaplasied cells that gets transmitted through subsequent growth promoter-mediated cell expansion, until the commitment of the metaplastic cell and those in its neighbourhood enabling the start of ‘novel’ cell-cycle patterns, cell multiplication, programmed differentiation, and control is needed to explain symmetry - a morphogenetic phenomenon that makes the insect-induced galls distinct from the bacteria-induced tumors.     

Cynipid galls: insect-induced modifications of plant development create novel plant organs

Cynipid gall formation is achieved by an insect–plant interaction whereby cynipid gallwasps redirect host‐plant development to form novel structures to protect and nourish the developing larvae. Work was carried out to investigate the molecular mechanisms involved in this interaction, and extend the understanding of plant tissue development. Cytological changes of the inner‐gall tissue throughout the development of several gall species was investigated and the developmental stages of gall formation defined, to reveal two different patterns of development followed by the galls tested. Fluorescent in situ hybridization demonstrated many of the inner‐gall cells to be polytenized. Comparisons between inner‐gall and non‐gall tissue protein signatures by Schönrogge et al. (Plant, Cell and Environment 23, 215–222, 2000) have demonstrated the variation between gall and non‐gall protein signatures, and identified a number of inner‐gall proteins. Further analysis of one of these inner‐gall proteins involved in lipid synthesis, putative biotin carboxyl carrier protein (BCCP), revealed differential expression throughout development, and showed this expression to be concentrated in the inner‐gall tissue in all the gall species tested.     

Exogenous phytohormones and the induction of plant galls by insects

The mechanism of gall induction by insects has remained elusive. Previous studies have met with limited success in attempting to induce galls by injection or application of chemical compounds. To determine whether an exogenous source of phytohormones plays a role in gall induction, we injected cytokinin (CK), auxin (IAA), gibberellic acid (GA), and abscisic acid (ABA) in various concentrations, ratios, and combinations into leaf petioles of Capsicum annuum L. cv. California Wonder (Solanaceae). We found that CK + IAA injections lead to gall-like growth in C. annuum. GA enhanced and ABA inhibited gall growth except in the presence of GA. Isopentenyl adenine (IP) was the most effective type of CK at inducing growth. Our work is consistent with the hypothesis that exogenous CK + IAA produced and supplied by insects leads to gall induction. We hypothesize that insects have obtained the capability to induce galls via acquisition of the biosynthetic pathways to produce IAA and trans-zeatin family CKs through microbial symbiosis or lateral gene transfer.     

On electric potential along the plant stem

Electric potential difference along the stem of Pelargonium zonale plant has been measured. Three relaxation time values have been established. They correspond to different types of interelectrode space polarization (ion, ion-molecular, combined molecular-associative).     

On the electric potential along the plant stem

The electric potential difference along the stem of a Pelargonium zonale plant has been measured. Three relaxation time values have been established. They correspond to different types of interelectrode space polarization (ion, ion-molecular, combined molecular-associative).     

Stem galls affect oak foliage with potential consequences for herbivory

Abstract.   1. On two dates, foliar characteristics of pin oak, Quercus palustris , infested with stem galls caused by the horned oak gall, Callirhytis cornigera , were investigated, and the consequences for subsequent herbivory assessed.
2. Second-instar caterpillars of the gypsy moth, Lymantria dispar , preferred foliage from ungalled trees.
3. Ungalled trees broke bud earlier than their galled counterparts.
4. Galled trees produced denser leaves with higher nitrogen and tannin concentrations, but foliar carbohydrates did not differ among galled and ungalled trees.
5. Concentrations of foliar carbohydrates in both galled and ungalled trees increased uniformly between the two assay dates. Nitrogen concentrations were greater in leaves from galled trees, and decreased uniformly in galled and ungalled trees over time. Foliar tannins were also greater in foliage from galled trees early in the season; however, foliar tannins declined seasonally in galled tissue so that by the second assay date there was no difference in tannin concentrations between galled and ungalled foliage.
6. In spite of differences in foliar characteristics, performance of older, fourth instar gypsy moth caterpillars did not differ between galled and ungalled trees.     

On the theory of action potential propagation in plant cells

The distribution of an electric field in plant cells and zooblasts has been investigated during propagation of the action potential. The behavior of ions in the cytoplasm and in the extracellular fluid has been described with the equations of electric charge motion in electrolytes. It has been shown that the action potential causes an electric potential change not only in the depth of the cytoplasm but also in the extracellular area far from the lipid bilayer. The biomembrane resistance has been expressed by physical parameters of a cell, such as ionic diffusion coefficient in fluid, Debye-Hückel radius, dielectric constant etc. The presence of breaks in the action potential diagrams has been explained as a result of insufficient resolving power of the measuring devices at the instant the sodium ion channels of the bilayer open.     

Structure-activity relationship of acetylenes from galls of Hedera rhombea as plant growth inhibitors

The structure-activity relationship of 12 isolated acetylenes from galls of Hedera rhombea (Araliaceae) induced by Asphondylia sp. (Cecidomyiidae) and their derivatives has been studied for the inhibition of the shoot and root growth of rice, perennial ryegrass, cockscomb, lettuce, and cress. Almost all acetylenes generally showed growth inhibitory activity. The diacetylenes exhibited higher activity than the monoacetylenes, suggesting that a conjugated diyne segment is essential for the activity. On the other hand, the acetylenes with a nonoxidated methylene group at C-8 showed stronger activity comparing with those possessing hydroxy and acetoxy groups at C-8. Furthermore, it has been demonstrated that the acetylenes bearing a terminal olefinic group at C-16, C-17 enhanced the activity. It is thus clarified that important sites for the activity of the acetylenes from galls of H. rhombea are a conjugated diyne and a terminal olefinic group connecting to the aliphatic chain and that less oxidated compounds show more activity.     

The influence of different nutrient levels on insect-induced plant volatiles in Bt and conventional oilseed rape plants

Transgenic Bt (expressing the cry1Ac endotoxin gene) and conventional oilseed rape plants grown in different soils were used to study nutrient uptake and emission of volatiles after herbivore damage. All plants were greenhouse-grown in soils representing low-, medium- and high-nutrient levels. The concentrations of N, P, K, Mg and Zn were significantly affected by the transgene, while the main effect of soil type appeared in N, P, Ca, Mg, B, Mn and Zn concentrations in the plants. Plants with four to five leaves were infested with the third instar larvae of Bt-susceptible Plutella xylostella for 48 h, and samples of volatiles were collected and analysed. In the first experiment, the soil nutrient level had a significant effect on the emissions of (Z)-3-hexen-1-ol, (Z)-3-hexenyl acetate, hexyl acetate, (E)-4,8-dimethyl-1,3,7-non-atriene (DMNT), beta-elemene, gamma-bisabolene, alpha-bisabolene and (E)-nerolidol. The induction of these volatiles was significantly higher in infested conventional plants grown at a high-soil nutrient level compared to infested conventional plants at a low-soil nutrient level. In the second experiment, the soil nutrient level had a significant effect on the emissions of (Z)-3-hexen-1-ol, (Z)-3-hexenyl acetate and beta-elemene and, again, this was significantly higher in infested conventional plants grown at high-soil nutrient levels in comparison with infested plants at a low-soil nutrient level. In both experiments, the transgene effect was significant on the emissions of DMNT and (E,E)-alpha-farnesene. The differences in emissions between the two separate experiments suggest that growth conditions (particularly daylength) and sampling procedure may affect the ratio of compounds detected in the emission blend, even though the response to herbivory, nutrient availability and the transgene were similar.     

Ant effects on three-trophic level interactions: plant, galls, and parasitoids

1. Ants have evolved mutualistic relationships with a diverse array of plant and animal species. Usually, the predatory/aggressive behaviour of ants near food sources can limit herbivore damage. 2. Galls of Disholcaspis edura on Quercus turbinella produce a secretion that is harvested by three species of ants (Formica neorufibarbis, Liometopium apiculatum, and Monomorium cyaneum) in the chaparral vegetation of Arizona, U.S.A. The study reported here provides evidence of a mutualistic relationship between these species of ants and the gall-forming wasp Disholcaspis edura. 3. An ant exclusion experiment showed that when ants tended galls, the rate of parasitism by Platygaster sp. on Disholcaspis edura was nearly halved in comparison to a treatment in which ants were excluded. 4. In the presence of ants, galls with the largest diameter suffered a lower mortality rate due to parasitoid attack than when ants were excluded. Thus, ant presence reduced the selective pressure imposed by Platygaster sp. on the galls with larger diameter.     

Lipids of oak galls

The nutrient tissues of oak galls accumulate a great amount of lipids. The neutral lipids (essentially triacylglycerol) are the most abundant storage form but some membrane lipids (mainly phospholipids) also occur. However, the galactolipids are very poorly represented. Among the fatty acids, oleic acid is predominant. These data correlate well with the cytological features of gall nutrient cells.     

The chemical composition of plant galls: are levels of nutrients and secondary compounds controlled by the gall-former?

The chemical composition of galled and ungalled plant tissue was compared in a series of experiments. Gall and adjacent plant tissue was analysed for 20 species of gall-former on 11 different plant species. There were clear differences between galled and ungalled tissue in levels of nutrients and secondary compounds. Gall tissue generally contained lower levels of nitrogen and higher levels of phenolic compounds than ungalled plant tissue. The gall tissue produced by the same plant in response to different species of gall-former differed in chemical composition, as did the gall-tissue from young and mature galls of the same species. The chemical differences between gall and plant tissues were studied in more detail in two field manipulations. Firstly, the seasonal changes in phenolic biosynthesis in Pontania proxima and P. pedunculi (Hymenoptera: Tenthredinidae) gall tissue were compared to those of their host plants, Salix alba and S. caprea. In both types of gall tissue, phenolic levels declined as the season progressed, but levels in the surrounding plant tissue increased. When the gall insects were killed with insecticide, phenolic levels in the galled tissue dropped to the same level as those in adjacent plant tissue. Secondly, the density of Cynips divisa (Hymenoptera: Cynipidae) galls on Quercus robur leaves was reduced by removing half the galls present, either those from the central region of the leaf or those from the edge. Decreasing gall density increased the size of the remaining galls and the weight of the insects, but these effects were most marked when the galls remaining were growing centrally on the leaf, i.e. when the galls from the edge had been removed. Decreasing gall density increased the nitrogen content of the remaining galls, again to a greater extent in galls growing centrally on the leaf. The results of these studies suggest that the levels of nutrients and secondary compounds in gall tissue are usually markedly different to those of surrounding plant tissue, and that gall-formers may produce species-specific and temporally variable changes in the chemical composition of gall tissue. Received: 7 July 1997 / Accepted: 29 September 1997     

On the steady-state electrical potential difference across the thylakoid membranes of chloroplasts in illuminated plant cells

The potential difference across the thylakoid membranes under steady-state saturating light conditions, measured with microcapillary glass electrodes, was found to be small as compared to the potential initially generated at the onset of illumination. This result is discussed to be in agreement with quantitative estimates on the approximate magnitudes of the potential generating electron flux through the photo-synthetic electron transport chain and of the potential dissipating ion fluxes across the thylakoid membrane under steady-state conditions. It is concluded that a pH gradient of approx. 3–3.4 units is built up in the light across the membrane. The negative diffusion potential associated with this gradient is suggested to cause the transient negative potential observed in the dark after illumination.