Rocio Cruz-Ortega

Effects of Allelochemical Stress Produced by Sicyos deppei on Seedling Root Ultrastructure of Phaseolus vulgaris and Cucurbita ficifolia

The allelopathic potential of the cucurbit Sicyos deppei has been reported. Aqueous leachates (1% w/v) of S. deppei significantly inhibited the radicle growth of several test plants. Root tips of treated plants were thicker, with a brownish color as compared to control roots. Light microscope observations of treated root tips of Phaseolus vulgaris showed that cells around the quiescent center appeared compressed. Tissues were disorganized and lacked evident cell differentiation. Cucurbita ficifolia did not show these differences in root morphology. Scanning and electron microscope observations showed that treated roots of both P. vulgaris and C. ficifolia had ultrastructural alterations of certain organelles, the plasma membrane, and cell walls. Root cap cells of treated roots showed amorphous and inactivated (nondividing) nuclei, mitochondria, and endoplasmic reticulum. Vacuoles in treated cells showed invaginations that may correspond to engulfing of damaged cellular components.

Allelochemical stress can trigger oxidative damage in receptor plants: mode of action of phytotoxicity.

Plants can interact with other plants through the release of chemical compounds or allelochemicals. These compounds released by donor plants influence germination, growth, development, and establishment of receptor plants; having an important role on the pattern of vegetation, i.e as invasive strategy, and on crop productivity. This phytotoxic or negative effect of the released allelochemicals (allelochemical stress) is caused by modifying or altering diverse metabolic processes, having many molecular targets in the receptor plants. Recently, using an aggressive and allelopathic plant Sicyos deppei as the donor plant, and Lycopersicon esculentum as the receptor plant, we showed that the allelochemicals released by S. deppei caused oxidative damage through an increase in reactive oxygen species (ROS) and activation or modification of antioxidant enzymes. Based on this study, we proposed that oxidative stress is one of the mechanisms, among others, by which an allelopathic plant causes phytotoxicity to other plants.

Screening for Effects of Phytochemical Variability on Cytoplasmic Protein Synthesis Pattern of Crop Plants

Crop plants have to cope with phytochemical variability along with other environmental stresses. Allelochemicals affect several cellular processes. We tested the effect of toxic aqueous leachates from Sicyos deppei, Acacia sedillense, Sebastiania adenophora, and Lantana camara on the radicle growth and cytoplasmic protein synthesis patterns of Zea mays (maize), Phaseolus vulgaris (bean), Cucurbita pepo (squash), and Lycopersicon esculentum (tomato). 2D-PAGE and gel scan densitometry analysis were used to detect differences in cytoplasmic root protein pattern expression. High-, medium-, and low-molecular-weight cytoplasmic proteins were affected by the different aqueous leachates. Crop plant responses were diverse, but in general, an increase in protein synthesis was observed in the treated roots. Maize was the least affected, but both the radicle growth and also the protein pattern of tomato were severely inhibited by all allelopathic plants. The changes observed in protein expression may indicate a biochemical alteration at the cellular level of the tested crop plants.

Allelochemical potential of Callicarpa acuminata

The allelochemical potential of Callicarpa acuminata (Verbenaceae) was investigated by using a biodirected fractionation study as part of a long-term project to search for bioactive compounds among the rich biodiversity of plant communities in the Ecological Reserve El Eden, Quintana Roo, Mexico. Aqueous leachate, chloroform-methanol extract, and chromatographic fractions of the leaves of C. acuminata inhibited the root growth of test plants (23–70%). Some of these treatments caused a moderate inhibition of the radial growth of two phytopathogenic fungi, Helminthosporium longirostratum and Alternaria solani (18–31%). The chloroform-methanol (1:1) extract prepared from the leaves rendered five compounds: isopimaric acid (1), a mixture of two diterpenols [sandaracopimaradien-19-ol (3) and akhdarenol (4)], α-amyrin (5), and the flavone salvigenin (6)]. The phytotoxicity exhibited by several fractions and the full extract almost disappeared when pure compounds were evaluated on the test plants, suggesting a synergistic or additive effect. Compounds (4), (5), and the semisynthetic derivative isopimaric acid methyl ether (2) had antifeedant effects on Leptinotarsa decemlineata. Compound 5 was most toxic to this insect, followed by (2), (4), and (6) with moderate to low toxicity. No correlation was found between antifeedant and toxic effects on this insect, suggesting that different modes of action were involved. All the test compounds were cytotoxic to insect Sf9 cells while (6), (4), and (1) also affected mammalian Chinese Hamster Ovary (CHO) cells. Compound 5 showed the strongest selectivity against insect cells. This study contributes to the knowledge of the defensive chemistry and added value of C. acuminata.

Conostegia xalapensis (Melastomataceae): an aluminum accumulator plant

In acidic soils, an excess of Al³⁺ is toxic to most plants. The Melastomataceae family includes Al-accumulator genera that tolerate high Al³⁺ by accumulating it in their tissues. Conostegia xalapensis is a common shrub in Mexico and Central America colonizing mainly disturbed areas. Here, we determined whether C. xalapensis is an Al accumulator, and whether it has internal tolerance mechanisms to Al. Soil samples collected from two pastures in the state of Veracruz, Mexico, had low pH and high Al³⁺ concentrations along with low Ca²⁺ levels. Leaves of C. xalapensis from pastures showed up to 19,000 mg Al kg⁻¹ DW (dry weight). In laboratory experiments, 8-month-old seedlings treated with 0.5 and 1.0 mM AlCl₃ for 24 days showed higher number of lateral roots and biomass. Pyrocatechol violet and hematoxylin staining evidenced that Al localized in epidermis and mesophyll cells in leaves and in epidermis and vascular pith in roots. Scanning electron microscope-energy dispersive X-ray microanalysis of Al-treated leaves corroborated that Al is in abaxial and adaxial epidermis and in mesophyll cells (31.2%) in 1.0 mM Al-treatment. Roots of Al-treated plants had glutathione reductase (EC 1.6.4.2) and superoxide dismutase (EC 1.15.1.1) activity higher, and low levels of O₂*⁻ and H₂O₂. C. xalapensis is an Al-accumulator plant that can grow in acidic soils with higher Al³⁺ concentrations, and can be considered as an indicator species for soils with potential Al toxicity.

Phytotoxic effects of Sicyos deppei (Cucurbitaceae) in germinating tomato seeds

The phytotoxic effect of allelochemicals is referred to as allelochemical stress and it is considered a biotic stress. Sicyos deppei G. Don (Cucurbitaceae) is an allelopathic weed that causes phytotoxicity in Lycopersicon esculentum, delaying seed germination and severely inhibiting radicle growth. This paper reports in in vitro conditions, the effects of the aqueous leachate of S. deppei-throughout tomato germination times-on (1) the dynamics of starch and sugars metabolism, (2) activity and expression of the cell wall enzymes involved in endosperm weakening that allows the protrusion of the radicle, and (3) whether abscisic acid (ABA) is involved in this altered metabolic processes. Results showed that S. deppei leachate on tomato seed germination mainly caused: (1) delay in starch degradation as well as in sucrose hydrolysis; (2) lower activity of sucrose phosphate synthase, cell wall invertase, and alpha-amylase; being sucrose phosphate synthase (SPS) gene expression down-regulated, and the last two up regulated; (3) also, lower activity of endo beta-mannanase, beta-1,3 glucanase, alpha-galactosidase, and exo-polygalacturonase with altered gene expression; and (4) higher content of ABA during all times of germination. The phytotoxic effect of S. deppei aqueous leachate is because of the sum of many metabolic processes affected during tomato seed germination that finally is evidenced by a strong inhibition of radicle growth.

Morphological variation in the flowers of Jacaratia mexicana A. DC. (Caricaceae), a subdioecious tree

The Caricaceae is a small family of tropical trees and herbs in which most species are dioecious. In the present study, we extend our previous work on dioecy in the Caricaceae, characterising the morphological variation in sexual expression in flowers of the dioecious tree Jacaratia mexicana. We found that, in J. mexicana, female plants produce only pistillate flowers, while male plants are sexually variable and can bear three different types of flowers: staminate, pistillate and perfect. To characterise the distinct types of flowers, we measured 26 morphological variables. Our results indicate that: (i) pistillate flowers from male trees carry healthy-looking ovules and are morphologically similar, although smaller than, pistillate flowers on female plants; (ii) staminate flowers have a rudimentary, non-functional pistil and are the only flowers capable of producing nectar; and (iii) perfect flowers produce healthy-looking ovules and pollen, but have smaller ovaries than pistillate flowers and fewer anthers than staminate flowers, and do not produce nectar. The restriction of sexual variation to male trees is consistent with the evolutionary path of dioecy from hermaphrodite ancestors through the initial invasion of male-sterile plants and a subsequent gradual reduction in female fertility in cosexual individuals (gynodioecy pathway), but further work is needed to confirm this hypothesis.

Aporphine alkaloid contents increase with moderate nitrogen supply in Annona diversifolia Saff. (Annonaceae) seedlings during diurnal periods.

Abstract Aporphine alkaloids are secondary metabolites that are obtained in low levels from species of the Annonaceae family. Nitrogen addition may increase the alkaloid content in plants. However, previous studies published did not consider that nitrogen could change the alkaloid content throughout the day. We conducted this short-term study to determine the effects of nitrogen applied throughout the diurnal period on the aporphine alkaloids via measurements conducted on the roots, stems and leaves of Annona diversifolia seedlings. The 60-day-old seedlings were cultured with the addition of three levels of nitrogen (0, 30 and 60 mM), and alkaloid extracts were analysed using high-performance liquid chromatography. The highest total alkaloid content was measured in the treatment with moderate nitrogen supply. Further, the levels of aporphine alkaloids changed significantly in the first few hours of the diurnal period. We conclude that aporphine alkaloid content increased with moderate nitrogen supply and exhibited diurnal variation.

Plants from the abandoned Nacozari mine tailings: evaluation of their phytostabilization potential

Phytostabilization is a remediation technology that uses plants for in-situ stabilization of contamination in soils and mine tailings. The objective of this study was to identify native plant species with potential for phytostabilization of the abandoned mine tailings in Nacozari, Sonora in northern Mexico. A flora of 42 species in 16 families of angiosperms was recorded on the tailings site and the abundance of the most common perennial species was estimated. Four of the five abundant perennial species showed evidence of regeneration: the ability to reproduce and establish new seedlings. A comparison of selected physicochemical properties of the tailings in vegetated patches with adjacent barren areas suggests that pH, electrical conductivity, texture, and concentration of potentially toxic elements do not limit plant distribution. For the most abundant species, the accumulation factor for most metals was <1, with the exception of Zn in two species. A short-term experiment on adaptation revealed limited evidence for the formation of local ecotypes in Prosopis velutina and Amaranthus watsonii. Overall, the results of this study indicate that five native plant species might have potential for phytostabilization of the Nacozari tailings and that seed could be collected locally to revegetate the site. More broadly, this study provides a methodology that can be used to identify native plants and evaluate their phytostabilization potential for similar mine tailings.

Plant-Mycorrhizae and Endophytic Fungi Interactions: Broad Spectrum of Allelopathy Studies

Plants may compete with one another for resources and they could also release allelochemicals that inhibit or stimulate the development and survival of neighboring plants and microorganisms. Arbuscular mycorrhizal fungi (AMF) and fungal endophytes are part of the microbial community of almost every plant; the composition of a plant microbial community is thought to affect plant fitness and physiology as well as their interactions with other plants. Arbuscular mycorrhizal fungi and endophytes could affect the interactions between invasive and native plants or crops; this may be due to modifications in soil microbial communities mediated by allelochemicals produced by those fungi and plants. This chapter discusses the possible allelopathic interactions between maize and Rottboellia cochinchinensis, as well as the possible interactions with their associated AMF. Other group of microorganisms considered here are endophytic fungi. This type of fungi produces allelochemicals with a great chemical diversity and biological activities. Interactions between plants and their associated microbial communities are mediated by allelochemicals; direct or indirect effects of these compounds on plants and microorganisms are considered within the spectrum of allelopathy.