Galls induced by a root-knot nematode in Petroselinum crispum (Mill.): impacts on host development, histology, and cell wall dynamics.

Infection by the root-knot nematode (RKN), Meloidogyne incognita, impacts crop productivity worldwide, including parsley cultures (Petroselinum crispum). Meloidogyne infection involves a complex relationship between the pathogen and the host plant tissues, leading to the formation of galls and feeding sites that disorganize the vascular system, affecting the development of cultures. Herein, we sought to evaluate the impact of RKN on the agronomic traits, histology, and cell wall components of parsley, with emphasis on giant cell formation. The study consisted of two treatments: (i) control, where 50 individuals of parsley grew without M. incognita inoculation; and (ii) inoculated plants, where 50 individuals were exposed to juveniles (J2) of M. incognita. Meloidogyne incognita infection affected the development of parsley, reducing the growth of some agronomical characteristics such as root weight and shoot weight and height. Giant cell formation was noticed at 18 days after inoculation, promoting disorganization of the vascular system. Epitopes of HGs detected in giant cells reveal the continuous capacity of giant cells to elongate under the stimulus of RKN, essential processes for feeding site establishment. In addition, the detection of epitopes of HGs with low and high methyl-esterified groups indicates the PMEs activity despite biotic stress.

Impact of Meloidogyne incognita (nematode) infection on root tissues and cell wall composition of okra (Abelmoschus esculentus L. Moench, Malvaceae).

Root-knot nematodes are endoparasites whose mature females lodge and grow inside the root of some cultivated plants, leading to losses in productivity. Herein, we investigated if the infection of okra, Abelmoschus esculentus (Malvaceae), promoted by the root-knot nematode Meloidogyne incognita (Meloidogynidae) changes some agronomic traits of the host plant, as well as the cell wall composition of the root tissues. The okra Santa Cruz 47® cultivar was infected with a suspension of 5000 M. incognita juveniles. The inoculated and non-inoculated okra plants were then submitted to morphological analysis at the end of experiment, as well as histological (at 4, 11, 18, 39, ad 66 days after inoculation) and immunocytochemical analysis (control and 66 days after inoculation). Root-knot nematode infection reduced the dry weight of the stem system but, unexpectedly, the number and weight of fruits increased. At 11 days after inoculation, we detected the presence of giant cells that increased in number and size until the end of the experiment, at 66 days after inoculation. These cells came from the xylem parenchyma and showed intense and moderate labeling for epitopes recognized by JIM5 and JIM7. The presence of homogalacturonans (HGs) with different degrees of methyl esterification seems to be related to the injuries caused by the nematode feeding activity and to the processes of giant cell hypertrophy. In addition, the presence of HGs with high methyl-esterified groups can increase the cell wall porosity and facilitate the flux of nutrients for the root-knot nematode.

Patterns of cell elongation in the determination of the final shape in galls of Baccharopelma dracunculifoliae (Psyllidae) on Baccharis dracunculifolia DC (Asteraceae).

Cell redifferentiation, division, and elongation are recurrent processes, which occur during gall development, and are dependent on the cellulose microfibrils reorientation. We hypothesized that changes in the microfibrils orientation from non-galled tissues to galled ones occur and determine the final gall shape. This determination is caused by a new tissue zonation, its hyperplasia, and relative cell hypertrophy. The impact of the insect's activity on these patterns of cell development was herein tested in Baccharopelma dracunculifoliae-Baccharis dracunculifolia system. In this system, the microfibrils are oriented perpendicularly to the longest cell axis in elongated cells and randomly in isodiametric ones, either in non-galled or in galled tissues. The isodiametric cells of the abaxial epidermis in non-galled tissues divided and elongated periclinally, forming the outer gall epidermis. The anticlinally elongated cells of the abaxial palisade layer and the isodiametric cells of the spongy parenchyma originated the gall outer cortex with hypertrophied and periclinally elongated cells. The anticlinally elongated cells of the adaxial palisade layer originated the inner cortex with hypertrophied and periclinally elongated cells in young and mature galls and isodiametric cells in senescent galls. The isodiametric cells of the adaxial epidermis elongated periclinally in the inner gall epidermis. The current investigation demonstrates the role of cellulose microfibril reorientation for gall development. Once many factors other than this reorientation act on gall development, it should be interesting to check the possible relationship of the new cell elongation patterns with the pectic composition of the cell walls.

Variation in the degree of pectin methylesterification during the development of Baccharis dracunculifolia kidney-shaped gall.

Insect galls may be study models to test the distribution of pectins and arabinogalactan-proteins (AGPs) and their related functions during plant cell cycles. These molecules are herein histochemically and immunocitochemically investigated in the kidney-shaped gall induced by Baccharopelma dracunculifoliae (Psyllidae) on leaves of Baccharis dracunculifolia DC. (Asteraceae) on developmental basis. The homogalacturonans (HGAs) (labeled by JIM5) and the arabinans (labeled by LM6) were detected either in non-galled leaves or in young galls, and indicated stiffening of epidermal cell walls, which is an important step for cell redifferentiation. The labeling of HGAs by JIM7 changed from young to senescent stage, with an increase in the rigidity of cell walls, which is important for the acquaintance of the final gall shape and for the mechanical opening of the gall. The variation on the degree of HGAs during gall development indicated differential PMEs activity during gall development. The epitopes recognized by LM2 (AGP glycan) and LM5 (1-4-ß-D-galactans) had poor alterations from non-galled leaves towards gall maturation and senescence. Moreover, the dynamics of pectin and AGPs on two comparable mature kidney-shaped galls on B. dracunculifolia and on B. reticularia revealed specific peculiarities. Our results indicate that similar gall morphotypes in cogeneric host species may present distinct cell responses in the subcelular level, and also corroborate the functions proposed in literature for HGAs.

The role of pectic composition of cell walls in the determination of the new shape-functional design in galls of Baccharis reticularia (Asteraceae).

The pectic composition of cell wall is altered during the processes of cell differentiation, plant growth, and development. These alterations may be time-dependent, and fluctuate in distinct regions of the same cell or tissue layer, due to the biotic stress caused by the activity of the gall inducer. Among the roles of the pectins in cell wall, elasticity, rigidity, porosity, and control of cell death may be crucial during gall development. Galls on Baccharis reticularia present species-specific patterns of development leading to related morphotypes where pectins were widely detected by Ruthenium red, and the pectic epitopes were labeled with specific monoclonal antibodies (LM1, LM2, LM5, LM6, JIM5, and JIM7) in distinct sites of the non-galled and the galled tissues. In the studied system B. reticularia, the epitopes for extensins were not labeled in the non-galled tissues, as well as in those of the rolling and kidney-shaped galls. The high methyl-esterified homogalacturonans (HGA) were labeled all over the tissues either of non-galled leaves or of the three gall morphotypes, while the intense labeling for arabinogalactans was obtained just in the rolling galls. The pectic composition of non-galled leaves denotes their maturity. The kidney-shaped gall was the most similar to the non-galled leaves. The pectic dynamics in the gall tissues was particularly altered in relation to low methyl-esterified HGA, which confers elasticity and expansion, as well as porosity and adhesion to cell walls, and are related to the homogenization and hypertrophy of gall cortex, and to translocation of solutes to the larval chamber. Herein, the importance of the pectic dynamics of cell walls to the new functional design established during gall development is discussed for the first time. The repetitive developmental patterns in galls are elegant models for studies on cell differentiation.

Cytological and histochemical gradients on two Copaifera langsdorffii Desf. (Fabaceae)--Cecidomyiidae gall systems.

Previous ultrastructural and histochemical analysis proposed patterns in the accumulation of substances in galls of Diptera: Cecidomyiidae in some plant species of the temperate region. Similar analyses were done to verify the conservativeness of these patterns in the Neotropical region, where a great number of species of Cecidomyiidae is responsible for a wide diversity of morphotypes. Two gall morphotypes induced by Cecidomyiidae in a unique host plant, Copaifera langsdorffii, were studied. The gradients of carbohydrates and the activity of invertases and acid phosphatases were similar, but the cytological gradients and distribution of proteins evidenced that the sites of the induction as well as the amount of neoformed tissues may be peculiar to each gall system. The production of lipids just in the secretory cavities either in the non-galled or galled tissues indicated a potentiality of the host plant which could not be manipulated by the galling insects. Further, the absence of nucleus in the nutritive tissue, an exclusive feature of the horn-shaped galls, indicates cell death attributed to the feeding habit of the galling herbivore.

Is the oxidative stress caused by Aspidosperma spp. galls capable of altering leaf photosynthesis?

The generation of ROS (reactive oxygen species) in plant galls may induce the degradation of the membrane systems of a plant cell and increase the number of plastoglobules. This numerical increase has been related to the prevention of damage to the thylakoid systems, and to the maintenance of photosynthesis rates. To investigate this hypothesis in gall systems, a comparative study of the ultrastructure of chloroplasts in non-galled leaves and in leaf galls of A. australe and A. spruceanum was conducted. Also, the pigment composition and the photosynthetic performance as estimated by chlorophyll fluorescence measurements were evaluated. The ultrastructural analyses revealed an increase in the number and size of plastoglobules in galls of both species studied. The levels of total chlorophylls and carotenoids were lower in galls than in non-galled tissues. The chlorophyll a/b ratio did not differ between the non-galled tissues and both kinds of galls. The values of maximum electron transport rate (ETR(MAX)) were similar for all the samples. The occurrence of numerous large plastoglobules in the galled tissues seemed to be related to oxidative stress and to the recovery of the thylakoid membrane systems. The maintenance of the ETR(MAX) values indicated the existence of an efficient strategy to maintain similar photosynthetic rates in galled and non-galled tissues.

Do Cecidomyiidae galls of Aspidosperma spruceanum (Apocynaceae) fit the pre-established cytological and histochemical patterns?

Cecidomyiidae galls commonly present a zonation of tissues with lignified cell layers externally limiting a reserve tissue and internally limiting a specialized nutritive tissue next to the larval chamber. The cytological aspects of this specialized tissue indicate high metabolic activity as well as carbohydrate accumulation. In Aspidosperma spruceanum-Cecidomyiidae gall system, ultrastructural and histochemical investigations corroborated this pattern and also revealed the storage of proteins in the nutritive cells. Reactive oxygen species (ROS), callose, and pectin accumulation were related to the feeding activity of the galling herbivore. Phosphorylase, glucose-6-phosphatase, acid phosphatases, invertases, and sucrose synthase activities were detected for the first time, in the Neotropical region, and discussed in relation to gall maintenance and the feeding activity of the Cecidomyiidae.