David Marín

Plant biocommunicators: their phytotoxicity, degradation studies and potential use as herbicide models

The development of new bioactive molecules with potential application in pharmacology and agriculture by using natural products as templates has been a widely used approach in the recent years. Focusing our attention in phytochemicals with potential application in agriculture, allelochemicals (natural plant toxins) have been deeply researched with the main purpose of finding phytotoxic substances to use as new herbicidal templates. The development of new analytical methodologies, improved bioassay techniques and a complete understanding of the allelopathic phenomenon will provide new tools for natural herbicide models development, in the context of the new approaches to integrated pest management. A deeper knowledge of the release and assimilation of allelochemicals, and the degradation phenomena associated to them will aid to discover new chemical structures with potential utility, in addition to a better understanding on the ecological interactions mediated by phytochemicals. The research on these topics, made with promising allelochemicals such as benzoxazinones and their degradation derivatives, the improvements on analytical design, the degradation mechanisms elucidation and the novel bioassay techniques recently developed, are revised herein.

Rediscovering the bioactivity and ecological role of 1,4-benzoxazinones

Compounds of the (2H)-1,4-benzoxazin-3(4H)-one class have attracted the attention of phytochemists since the first isolation of 2,4-dihydroxy-2H-1,4-benzoxazin-3(4H)-one (DIBOA) and 2,4-dihydroxy-7-methoxy-(2H)-1,4-benzoxazin-3(4H)-one (DIMBOA). Extensive research has been carried out on the isolation and synthesis of these materials as well as on the dynamics of their degradation in different systems. This has led to the discovery of a wide variety of compounds that are of high interest from the point of view of phytotoxic, antifungal, antimicrobial, and antifeedant effects among others. The potential application of benzoxazinones and their derivatives as leads for natural herbicide models is a topic of current interest. Furthermore, the importance of degradation on the ecological behaviour of benzoxazinone-producing plants is also being realised, and proposals concerning the role of the degradation products in chemical defence mechanisms have been put forward. There is also increasing interest in the improvement of analytical methodologies, and ecotoxicologic effects, toxicity on target and non-target organisms, and degradation kinetics are also being addressed. The development of new phytotoxicity bioassay techniques represents one of the most important breakthroughs in this respect. Moreover, benzoxazinones and some of their derivatives have been employed in the development of pharmaceuticals. The versatility of the benzoxazinone skeleton, in addition to its relative chemical simplicity and accessibility, makes these chemicals amongst the most promising sources of bioactive compounds that are natural in origin.

Synthesis of heliannane skeletons. Facile preparation of (±)-heliannuol D☆

Heliannuol D is a natural product with a 7,10-heliannane skeleton, isolated from Helianthus annuus. It has been synthesized in eight steps, in good yield, using a new biomimetic method. Key steps were a Fries rearrangement, a Grignard reaction and, finally, a base catalyzed cyclization.

Evidence for an Allelopathic Interaction Between Rye and Wild Oats

Allelopathy is a biological phenomenon in which an organism produces one or more biochemicals that influence the growth, survival, and reproduction of other organisms. Allelopathy has been the subject of a great deal of research in chemical ecology since the 1930s. The characterization of the factors that influence this phenomenon has barely been explored, mainly due to the complexity of this area. The main aim of the research carried out to date has been to shed light on the importance of these interactions in agroecosystems, especially in relation to the interactions between crops and weeds. Herein we report the characterization of a complete allelochemical pathway involving benzoxazinones, which are known to participate in allelopathic plant defense interactions of several plants of high agronomic interest. The production of the defense chemicals by a donor plant (crop), the route and transformations of the chemicals released into the environment, and the uptake and phytotoxic effects on a target plant (weed) were all monitored. The results of this study, which is the first of its kind, allowed a complete dynamic characterization of the allelopathic phenomenon for benzoxazinones.

Interactions of Bacillus mojavensis and Fusarium verticillioides with a Benzoxazolinone (BOA) and its Transformation Product, APO

The benzoxazolinones, specifically benzoxazolin-2(3H)-one (BOA), are important transformation products of the benzoxazinones that can serve as allelochemicals providing resistance to maize from pathogenic bacteria, fungi, and insects. However, maize pathogens such as Fusarium verticillioides are capable of detoxifying the benzoxazolinones to 2-aminophenol (AP), which is converted to the less toxic N-(2-hydroxyphenyl) malonamic acid (HPMA) and 2-acetamidophenol (HPAA). As biocontrol strategies that utilize a species of endophytic bacterium, Bacillus mojavensis, are considered efficacious as a control of this Fusarium species, the in vitro transformation and effects of BOA on growth of this bacterium was examined relative to its interaction with strains of F. verticillioides. The results showed that a red pigment was produced and accumulated only on BOA-amended media when wild type and the progeny of genetic crosses of F. verticillioides are cultured in the presence of the bacterium. The pigment was identified as 2-amino-3H-phenoxazin-3-one (APO), which is a stable product. The results indicate that the bacterium interacts with the fungus preventing the usual transformation of AP to the nontoxic HPMA, resulting in the accumulation of higher amounts of APO than when the fungus is cultured alone. APO is highly toxic to F. verticillioides and other organisms. Thus, an enhanced biocontrol is suggested by this in vitro study.

First total synthesis of (±)-helibisabonol A

Helibisabonol A is a new sesquiterpene with phytotoxic activity isolated from Helianthus annuus leaves. (±)-Helibis- abonol A has been synthesized as an approach to the natural product in the search for new herbicide models. Herein, we report the first total synthesis for this molecule from methylhydroquinone in six steps. The key steps of this synthesis are a Fries rearrangement, a Grignard reaction and a catalytic hydrogenation. The synthesis was carried out with high yield and in an easy to scale manner.

Combined strategy for phytotoxicity enhancement of benzoxazinones.

Fifteen new derivatives of D-DIBOA, including aromatic ring modifications and the addition of side chains in positions C-2 and N-4, were synthesized and their phytotoxicity, selectivity, and structure-activity relationships evaluated. The most active compounds among the derivatives at the C-2 position were 6-Cl-2-Et-D-DIBOA and 6-F-2-Et-D-DIBOA. Of the derivatives at N-4, the most active compounds were 6-Cl-4-Pr-D-DIBOA and 6-Cl-4-Val-D-DIBOA. These four compounds showed high levels of inhibition in root length at very low concentrations in all species. The most remarkable result is the 70% inhibition observed for the root length of cress at 100 nM caused by the latter two compounds. These results support our previous research and conclusions regarding the steric, electronic, and solubility requirements to achieve the maximum phytotoxic activity.

Modified benzoxazinones in the system Oryza sativa-Echinochloa crus-galli: an approach to the development of biorational herbicide models.

The utility of benzoxazinones and some of their synthetic derivatives in the search for new leads for herbicide model development has been explored. The work described focuses on obtaining derivatives that present selectivity in the system Oryza sativa- Echinochloa crus-galli. To achieve this goal the influence of lipophilicity in this system has been studied by preparing 14 ester derivatives at the N-4 position of D-DIBOA along with other compounds with different functionalization and chain lengths at position C-2. These compounds have been tested in the aforementioned system, and the dose-response profiles have been compared. The most active compound was 2-ethyl-4-hydroxy-(2H)-1,4-benzoxazin-3(4H)-one, which presented higher selectivity than the specific herbicide Cotanil-35. These results confirm the potential of D-DIBOA as a lead herbicide for the control of Echinochloa spp. in rice crops.

Aromatic-ring-functionalised benzoxazinones in the system Oryza sativa–Echinochloa crus-galli as biorational herbicide models

BACKGROUND Barnyardgrass, Echinochloa crus-galli (L.) Beauv., is one of the most problematic weeds occurring in rice crops. Although efficient chemical control is provided by herbicides available on the market, resistant biotypes provoked by pressure selection have appeared in recent times. This emphasises the need for alternative treatments in which herbicidal compounds from a natural origin could be included. RESULTS A number of chemicals with a [2H]-1,4-benzoxazin-3(4H)-one (D-DIBOA) skeleton were tested on this weed, and also in rice, in order to achieve an optimal lead for herbicide composition development by taking into consideration phytotoxic effects and selectivity on the weed. 6-Cl-D-DIBOA causes the same effect as the commercial herbicide propanil at a concentration 15 times lower, while 6-F-D-DIBOA causes this inhibition at a dose 30 times lower. The phytotoxicities caused by 8-Cl-D-DIBOA (IC50 = 44 microM, R2 = 0.866) and 7,8-diF-D-DIBOA (IC50 = 52 microM, R2 = 0.9067) are also remarkable. 8-Cl-D-DIBOA was the compound that presented the highest selectivity on Echinochloa crus-galli. The structural requirements for optimal phytotoxicity and selectivity were elucidated by means of QSAR methodology, considering electronic and steric factors. One of the most important descriptors influencing the bioactivity was the dipole moment modulus. This was successfully correlated by employing a second-order polynomial model. CONCLUSION The in vitro phytotoxic profiles and selectivities shown for these chemicals make them truly promising candidates for higher-level studies. 6F- and 6Cl-D-DIBOA, for their high phytotoxicities, and 8-Cl-D-DIBOA, because of its high selectivity, were found to be the most interesting compounds from this point of view.

Soil biodegradation of a benzoxazinone analog proposed as a natural products-based herbicide

AimsBenzoxazinones with the 4-hydroxy-(2H)-1,4-benzoxazin-3(4H)-one skeleton have been proposed as potentially successful models for the development of novel design leads. D-DIBOA has proven to be the most promising base structure in the search for novel herbicide models based on the benzoxazinone skeleton. The biodegradation dynamics of D-DIBOA in soil are therefore relevant and are the subject of this study.MethodsA previously optimized methodology for the assessment of biodegradation has been applied for the first time to a synthetic benzoxazinone.ResultsBiodegradability is a characteristic of natural benzoxazinones and a safety requirement for the development of herbicidal chemicals. The biodegradation phenomenon and its consequences for the development of new herbicide models are discussed. The half-life determined for D-DIBOA was much higher than those previously reported for the natural products DIBOA, DIMBOA and their benzoxazolinone derivatives.ConclusionsThis finding, together with its previously described potent phytotoxicity, suggests that D-DIBOA is a useful candidate for novel herbicide model development. The lactam D-HBOA, which is slightly less phytotoxic than its precursor, was discovered to be the first and principal metabolite resulting from D-DIBOA degradation.