WO2021118333A1

WO2021118333A1 - Fungal strains of the genus beauveria andcompositions containing same for the biological control of pest insects

Info

Publication number: WO2021118333A1
Application number: PCT/MX2020/000050
Authority: WO
Inventors: Adriana Rosalía GIJÓN HERNÁNDEZ; Guillermo SÁNCHEZ MARTÍNEZ; Saúl SANTANA ESPINOZA
Original assignee: Instituto Nacional De Investigaciones Forestales, Agrícolas Y Pecuarias (Inifap)
Priority date: 2019-12-11
Filing date: 2020-12-11
Publication date: 2021-06-17
Also published as: MX2019014970A

Abstract

The present invention relates to new isolated strains of the genus Beauveria, wherein Beauveria bassiana has accession number CM-CNRG TB96 and Beauveria sulfurescens has accession number CM-CNRG TB97, capable of actively infecting and causing high pathogenity in the biocontrol of pest insects. Additionally, the present invention provides an effective and efficient composition that includes at least one isolated strain of Beauveria bassiana and/or Beauveria sulfurescens for the prevention and/or biological control of a heterogeneous population of wood pests of the family Curculionidae.

Description

Strains of the Beauveria genus fungus and compositions containing them for the biological control of pest insects

FIELD OF THE INVENTION

The present invention belongs to the Forestry sector, particularly it refers to new isolated strains of Beauvería bassiana and Beauveria sulfurescens with the ability to actively infect and cause high pathogenicity in the biocontrol of pest insects. Additionally, the present invention provides compositions that include at least one isolated strain of an entomopathogenic fungus of the genus Beauveria for the prevention and / or control of forest pests of bark beetles of the Curculionidae family, especially to prevent, reduce or control biologically to heterogeneous populations of bark beetles causing damage to conifers.

BACKGROUND

Conifers constitute a group of plants known in the world, being of great relevance due to their provision of resources and environmental services for humans. Mexico has the greatest diversity of species, more than 91 species known and distributed in the mountainous forests of the Sierra Califomiana, Sierra Madre Occidental, Sierra Madre Oriental, Sierra Madre del Sur and Sierra Madre de Chiapas, of the 575 conifers that exist Worldwide, among these the families Pinaceae, Cupressaceae, Fagaceae, Obaceae and Ulmaceae have been reported. Currently its growing demand, conservation and maintenance represent economic value and benefit for the sustainability of future generations. However, conifers present variable disturbance regimes, due to the interactions of climate change and an imbalance of abiotic agents in conifers [Gemandt D. etal. Biodiversity of Pinophyta (conifers) in Mexico. Mexican Journal of Biodiversity. (2014); 85 (S): 126-133], One of the main negative impacts on conifers is due to the synergy between climate change and the favoring of aggressive agents such as some species of herbivorous beetles, among these more generally by bark beetles, which, from small outbreaks or large large-scale infestations, have caused an increase in the mortality of trees and shrubs, and consequently the alteration of ecosystems, being in Mexico the second agent that most affects temperate dima forests and cold temperate climates due to the magnitude of the affected area and the number of damaged trees . Around the world, more than 3,000 species of bark beetles have been described, approximately 870 species are distributed in Mexico. Among the main species of bark beetles are beetles of the Curculionidae family such as Dendroctonus spp., Hylesinus spp., Ips spp., Orthotomicus spp., Pityophthiorus spp., Phloeosinus spp., Pseudips spp., Pseudopityppthoph. ., and Pseudohylesinus spp. [Del-Val Ek. et al. Barking insects (coleoptera: curculíonidae) and dimatic change: customs problems and prospects in temperate forests. Specialized Magazine in Chemical-Biological Sciences. (2017); 20 (2); 53-60].

Taking into consideration the registry of notifications of forest sanitation due to bark insect infestation issued by SEMARNAT, an increase in risk areas has been identified in recent years. During 2016, the high-risk states consisted of Michoacán and Oaxaca, and with a moderate risk the states of Baja California Norte, Sonora, San Luis Potosí, Puebla, Guerrero and Chiapas. However, for the year 2019 these areas have shown an increase, representing a very high risk in the states of Michoacán, Oaxaca, Guerrero, Jalisco, Querétaro, San Luis Potosí, Chiapas and Nayarit, while the states of Puebla, Mexico , Veracruz, Hidalgo, Durango, T amaulipas, Morelos, Guanajuato, Mexico City, Chihuahua, Zacatecas, Nuevo Leon, Tlaxcala, Baja California, Coahuila, Colima and Sinaba at high risk. Said species of bark beetles are massively affecting and destroying a large part of the conifers, causing loss of production of forest resources and services, which attributes one of the country's environmental, economic and social losses. In order to implement the necessary measures to avoid damage to forest resources and also economic effects, the Ministry of the Environment and Natural Resources published NOM-019-SEMARNAT-2017 that establishes the technical guidelines for the prevention, combat and control of insects. debarkers. The first prevention action refers to monitoring in order to act in a timely manner and reduce impacts. Likewise, various techniques and control methods were included to actively reduce or eradicate outbreaks or their expansion of pests, among these, physical-mechanical methods such as sanitation cuts or the use of traps by means of attraction pheromones, as well as Also Ice chemical methods such as control by means of chemical or semi-chemical insecticides, however, not all pest species can apply the same control method. On the other hand, these practices turn out to be limiting to the stage of development of the pest, accessibility of the place and volume of the wood or infested area, and in some cases the use of these products generates resistance to insecticides or the control is unspecific, eliminating other favorable organisms of the ecosystem, therefore, it is necessary to implement different alternatives. This implies multiple treatments that requires monitoring by highly trained personnel and the repetitive application of chemical compounds, which is an ineffective measure for a population and its characteristics of the specific pest to be mitigated.

In urban environments, there is a very varied fauna that exerts both beneficial and harmful effects, therefore rejection situations can occur without distinguishing clearly between what we could call harmful fauna (mainly pests) and beneficial fauna (in some cases also the origin of some health problems such as phobias).

High population density, overcrowded conditions and inadequate sanitary facilities create an environment highly polluted by human and animal waste. This medium favors the proliferation and spread of diseases and favors infestation with mosquitoes, rodents, cockroaches, house flies, fleas, lice and many other arthropod vectors of diseases and pests. The goal of urban pest control is to improve the well-being of urban residents and reduce communicable diseases.

Currently, the use of biological controls for the management and control of agricultural, forest or urban pests has been investigated, that is, the use of organisms susceptible to a pathogenic organism, which represents an accessible and sustainable solution for the environment. Among the main microorganisms used, a potential has been demonstrated in the biological control of insects such as bacteria, viruses, fungi, nematodes and protozoa. The use of pathogenic microorganisms has taken on increasing interest as a biological control system for agricultural and forestry practice, among these bacteria such as Badllus thuringinensis, and fungi of species such as Beauvería bassiana, Metarhizium anisoplae, Langenedium giganteum, Lecanicillium lecanii and Paecelomyces stand out fumosomseus.

The National Center for Disciplinary Research in Conservation and Improvement of Forest Ecosystems (CENID-COMEF) of the National Institute of Forestry, Agricultural and Livestock Research (INIFAP), in the areas of research and innovation, have presented advances and results that have been developed in research projects in the areas of forest health. Among these investigations, multiple projects have been developed for the generation of new native isolates with high pathogenicity in biocontrol and the optimization of a composition for the prevention and / or biological control of pest insects. In the State of the Art, different strains and compositions containing an entomopathogenic fungus have been released for the control of various insects. These include the following:

MX / a / 2011/001033 that provides an isolate of Beauveria bassiana (strain No. 20763), as well as a useful composition for the control of pome fruits affected by the larva of the moth.

MX / a / 2014/014283 that provides an isolate of Beauveria bassiana (Bals. Vuill strain), as well as a useful composition for the control of fruits affected by the coffee bean borer.

MX / a / 2015/010030 that provides an insecticidal composition comprising a Beauveria bassiana strain (ABNBbl 01 regional strain), diatomaceous earth, sugars and dehydrated moringa vegetable leaf powder, where the composition is useful for the control of plants affected by whiteflies.

MX / a / 2016/005754 that provides a biopesticidal composition, characterized in that it comprises an agriculturally suitable carrier, a pesticidally effective amount of at least one fungal pesticide such as Beauveria bassiana, at least one sorbitan fatty acid surfactant, and at least one surfactant of sorbitol ethoxylate ester, where the composition is useful for the control of plants that damage agricultural crops affected by pests such as mites and ticks.

MX / a / 2017/003846 that provides a pesticidal composition comprising a fungal entomopathogenic fungus such as Beauveria bassiana (strain ATCC-74040), in combination with a natural pesticidal neem oil, where the composition is useful for the control of plants or agriculture affected by insects such as arthropods. Optionally said compositions contain surfactants, stabilizers, preservatives and / or other appropriate vehicles for the stability, delivery or application of the composition. Said compositions existing in the state of the art have allowed the isolation of specific strains and improved compositions for the prevention and control of agricultural pests. However, said compositions include Beauvería bassiana strains from isolates from various isolation sources such as agricultural pests, mainly arthropod insects of the Phylum Arthropoda with specific activity. Although Beauvería bassiana is an entomopathogenic fungus that causes epizootics, the generation of new native strains with high pathogenicity is necessary for its use as biocontrol of insect pests, mainly agricultural and forest pests.

Zhang Long et al. [Evaluation of Beauvería bassiana (Hyphomycetes) isolates as potential agents for control of Dendrodonus valens. Insect Science. (2011); 18: 209-216] have studied biological characteristics and virulence of Beauvería bassiana isolates, identifying isolates with high efficacy and virulence to be used as a potential sustainable biological control of the red turpentine beetle (Dendrodonus valens). These beetles are widely distributed in North America and China, mainly affecting coniferous species within their range.

However, the search for isolated native fungi with broad insecticidal activity for the prevention and control of pest insects is of interest, these species must provide high pathogenicity for biological control as alternatives to chemical insecticides intended to control pest species. agricultural and forestry. On the other hand, strains of entomopathogenic fungi are potential biological control agents, due to the production of toxins and their potential to damage the forest pest, however, the use of a synergistic composition containing genetically similar strains of the genus Beauvería can confer an antagonistic or synergistic effect for the biological control of pests, therefore, the mixture of the two strains must confer compatibility properties to generate dual germination and increase the tolerance range to abiotic factors. Additionally, said mixture of the strains in the synergistic composition must be capable of improving the efficacy for its use as biological control of heterogeneous populations both in vitro and in situ. Likewise, the generation of a composition must provide the components and conditions necessary for the growth, adhesion, penetration, and invasion of the entomopathogenic fungus to achieve the biological control of the forest pest. It should be noted that the efficacy and efficiency are not dependent on the synergistic combination in vitro of the isolated strains, since a composition can be affected due to abiotic field factors, such as relative humidity, precipitation and solar rays, for example. which, the composition as a whole, must ensure protection against abiotic factors to promote success in biological control. It is well known that climatic factors such as temperature, humidity and precipitation are parameters that can affect the type of interaction of the entomopathogenic fungus and the stability of the composition. Therefore, there has been no teaching or suggestion in the prior art, native isolates of Beauveria species and compositions with improved efficacy and efficiency characteristics for use as a bioinsecticide for the prevention and control of pest insects.

BRIEF DESCRIPTION OF THE INVENTION

New isolates of Beauveria bassiana with accession number CM-CNRG TB96 and Beauveria suifurescens with accession number CM-CNRG-TB97 are disclosed. Said strains provide high pathogenicity in the biocontrol of insect pests, including different species of arthropod insects, said isolated strains of the genus Beauveria are included in a composition that provides a synergistic effect for the biocontrol of one or more species of forest and agricultural pests in any of their habitats in urban and rural areas. In another embodiment, effective and efficient compositions are provided for the prevention and / or control of forest pests of the Curculionidae family, characterized in that the composition comprises at least one isolate of an entomopathogenic fungus selected from the group consisting of Beauveria bassiana (no. Of CM-CNRG TB96 accession) and Beauvería sulfurescens (CM-CNRG-TB97 accession no.) and combinations thereof; b) a natural substrate; c) a linear anionic surfactant; d) a soluble extract of brown seaweed; e) a plant extract of Moringa oleifera; and f) an acceptable carrier. In particular, the composition and its components provide the nutritional requirements and conditions necessary to improve the growth, adhesion, penetration and invasion of the entomopathogenic fungus in order to produce a biological control of the forest pest.

In addition to the above, the researchers have surprisingly developed a composition that, with the synergistic combination of the isolated strains, provides broad protection to coniferous species against the attack of heterogeneous populations of forest pests. Such findings provide an effective and efficient strategy for the prevention and / or control of forest pests of bark beetles of the Family Curculionidae.

This is proven with in situ tests, where the composition that includes a synergistic combination of the isolated strains of the genus Beauveria provides high mortality with abiotic factors both in vitro and in situ, against heterogeneous populations of pest insects.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1. Isolated from the strains in culture medium: a) Isolated strain of Beauveria bassiana:

White cottony growth; b) Isolated strain of Beauveria sutfurescens: Powdery white cobr growth.

Figure 2. Differences in the nucleotide sequences of the isolated strains of the genus Beauveria obtained from a native pest insect host from the database of the Genbank, with respect to the sequence of the Beauvería bassiana strain called GIINIF-15.

Figure 3, Differences in the nucleotide sequences of the isolated strains of the genus Beauvería obtained from a native pest insect host from the Genbank database, with respect to the sequence of the Beauvería suifurescens strain called

GIINIF-17.

Figure 4. Graph of the comparative evaluation of the mortality percentage of the Beauvería isolates in pest insects of the genus Dendrodonus spp.

Figure 5, Graph of the comparative evaluation of the percentage of mortality of the isolated strains of Beauvería in pest insects of the genus Ips spp.

Figure 6. Graph of the effect of different treatments on the mortality of bark beetles with respect to time. Where: H = Entomopathogenic fungus of Beauvería bassiana ', LV = Linear anionic surfactant agent; Diatomaceous earth; 103 = Aqueous extract of Moringa oleifera; 8D35 = Composition of the present invention. Cool letters are shared when there are no significant differences according to Tukey's test

(P <0.001).

Figure 7. Graph of the evaluation of the percentage of mortality in cockroaches of the composition of the present invention.

Figure 8. Graph of the Effect of the different treatments on the mortality of bark beetles with respect to time. Where: COT = Control group; 8D35 =

Composition of the present invention; MCB = Commercial composition of biological control. Equal letters are shared when there are no significant differences according to Tukey's test (P <0.001). Figure 9. Photograph of in situ validation of the composition of the present invention in species of bark beetle (Dendroctonus mexicanus) micosado.

OBJECT OF THE INVENTION

The object of the present invention are new isolated strains of Beauvería bassiana with accession number CM-CNRG TB96 and Beauvería sulfurescens with accession number CM-CNRG-TB97 with insecticidal activity. These strains provide a high mortality against pest insects and with a higher level of pathogenicity than the already known strains.

Another object of the present invention is a composition for the prevention and / or control of forest pests of bark beetles of the Curculionidae family, wherein the composition is characterized in that it comprises at least one isolate of an entomopathogenic fungus selected from the group consisting of Beauvería bassiana (Accession No. CM-CNRG TB96) and Beauvería sulfurescens (Accession No. CM-CNRG-TB97) and combinations thereof; b) a natural substrate; c) a linear anionic surfactant; d) a soluble extract of brown seaweed; e) a plant extract of Moringa oleifera: and f) an acceptable carrier. Said compositions provide efficacy and efficiency to prevent, reduce or biologically combat populations of bark beetles that cause damage to conifers.

DETAILED DESCRIPTION OF THE INVENTION

The detailed description set forth below is intended as a description of the presently preferred embodiments of the invention and is not intended to represent the only forms and applications in which the present invention may be reproduced or employed in practice, and as such may be used, incorporated or applied to other species of pests for the purpose and scope of the present invention in a non-limiting manner. The present invention provides new isolated strains of an entomopathogenic fungus belonging to the genus Beauveria. The entomopathogenic fungus as presented in the invention is a native species of the Beauveria bassiana species that causes a disease in various species of pest insects, which include species of insects such as the order Diptera (mosquitoes), Diptyoptera (cockroaches), Coleoptera. (beetles), Hemiptera (whitefly), Lepidoptera (worms) and / or Orthoptera (locusts), but not limited to other pests and their stages of development such as egg, larva, pupa and adult insect.

As used in the present description, "pests" is any organism or species of arthropod insects that can cause mechanical or physiological damage to plant species or that man considers harmful to his person, his property or the environment. In the first place, the Agricultural pests comprise one or more populations of phytophagous insects that cause damage to crop plants or agricultural products. Second, forest pests comprise one or more populations of phytophagous insects that cause damage to trees, forests in an ecosystem, or forest products. Finally, urban pests comprise one or more polyphagous insect populations.

Additionally, the present invention also provides a composition that includes at least one strain of entomopathogenic fungus belonging to the genus Beauveria, from which at least one of the species is selected: B. bassiana and B. sulfurescens for the prevention and / or control of Forest pests of the Curculionidae family, especially to prevent, reduce or biologically combat populations of bark beetles that cause damage to conifers. Said compositions of the invention include adjuvants or auxiliaries that allow them to improve their efficacy and efficiency for their use in the biological control of various species of conifers. As used in the present description, "conifers" are trees or small shrubs of the class conferopsida, whose reproductive structures are called cones or more commonly pinecones, for example, conifers include species such as cypresses, pines, firs, picaceae, sabines, oyameles junipers, ash trees, elms, but not limited to certain forest species with rough or smooth bark, in large, thick plates with fissures or in long, thin strips.

For the purposes of the present invention, the term "conifers" includes different species recognized by those skilled in the art as species of the Family Pinaceae, Cupressaceae, Fagaceae, Oleaceae and Ulmaceae.

In a preferred embodiment of the invention, the species of the Pinaceae family include pine, fir, and fir, but are not limited to genera of Pinus spp. , Abies spp. and Pseudotsuga spp. In an even more preferred embodiment, they include the genus Juniperus spp., Selected from at least one of the following coughs P. arizonica, P. ayacahuite, P. cembroides, P. chihuahuana, P. contorta, P. cooperi, P, culminicola, P . devoniana, P. douglasiana, P. duranguensis, P. edulis, P. engeimanni, P. esteveni, P. flexilis, P. georginae, P. greggii, P. hartwegii, P. herrerai, P. jaliscana, P. jeffreyi , P. lagunae, P. lawsoni, P. biophylla, P. longaeva, P. lumholtzii, P, luzmariae, P. maximartinezzi, P. mañminoi, P. michoacana, P. montezumae, P. monophylla, P. murícata, P . oocarpa, P. patula, P. pnceana, P. ponderosa, P. pringlei, P. pseudostrobus, P. quadrifolia, P. radíate, P. rudis, P. rzedowskii, P. tecunumaníi, P. teocote and P. yecorensis ; of the genus Abies spp., selected from at least one of the following: A. concolor, Aduranguensis, A.flinckii, A.hidalgensis, A.hickelli, Amexicana, Aoaxacana, Areligbsa and A. vejarii; of the genus Pseudotsuga spp., selected from at least one of the following P. flahaulti, P. guinieri, P. macrolepis, P. menziesii and P. rehderi.

In a preferred embodiment of the invention, the genera of the Cupressaceae family include juniper, sabino, cypress and ahuehuete, but are not limited to genera of Juniperus spp., Cupressus spp. and Taxodium spp. In an even more preferred embodiment, they include from the genus Juníperus spp., Selected from at least one of the following: J. angosturana, J. arizonica, J. ashei, J. califomica, J. comitana, J. deppeana, J. flaccida, J .jaliscana, J. martinezii, J. montícola, J. saltillensis, J. scopulorum, J. standleyi and J. zanonir, of the genus Cupressus spp., selected from at least one of the following C. arizonica, C. benthamii, C guadalupensis, C. iusitanica and C. stephensonii, · of the genus Taxodium spp., selected from at least one of the following species T. mucronatum and T. huegelii.

In a preferred embodiment of the invention, the genera of the Fagaceae family include oak and oak, but are not limited to genera of Quercus spp. In an even more preferred embodiment, they include the genus Quercus spp. , selected from at least one of the following coughs Q. acutifolia, Q. affinis, Q. candicans, Q. castanea, Q. coccolobifolia, Q. conspersa, Q. convallata, Q. crassifolia, Q. crassippes, Q. crispipillis, Q . durifolia, Q. eduardii, Q. eugeniaefolia, Q. excelsa, Q. glabrescens, Q. giaucoides, Q. grísea, Q. laeta, Q. laurina, Q. magnoliifolia, Q. mexicana, Q. obtusata, Q. peduncularis , Q. potosina, Quercus resinosa, Q. rugosa, Q. scytophyllia, Q. sideroxyila, Q. skinneri, Q. urbanii and Q. uxoris.

In a preferred embodiment of the invention, genera of the family Oleaoeae include ash, but are not limited to genera of Fraxinus spp. In an even more preferred embodiment they include from the genus Quercus spp., Selected from at least one of the following coughs F. americana, F. cuspidata, F. excelsior, F. omus, F. pennylvannica and F. uhdei. In a preferred embodiment of the invention, genera of the family Ulmaceae include elms, but are not limited to genera of Ulmus spp. In an even more preferred embodiment, they include the genus Quercus spp. , selected from at least one of the following coughs U. mexicana, U. parviflora and U. pumila. For the purposes of the present invention, the term "pests" includes different species of phytophagous insects recognized by those skilled in the art, among these it is selected from the group consisting of Coleoptera (beetles and weevils), Dermáptera (earwigs), Diptera (mosquitoes). ), Dictyoptera (cockroaches), Embioptera (weavers), Hemiptera (chiches, aphids, aphids), Hymenoptera (ants), Isoptera (termites), Lepidoptera (moths), Orthoptera (lobsters and crickets), Plecoptera (stone flies), Phithiraptera (lice, totolates), Psocoptera (book lice), Siphonaptera (fleas), Thysanoptera (thrips), but not limited to certain species of insects that cause damage to components in agricultural and forest ecosystems.

For the purposes of the present invention, the term "forest pests" includes different species recognized by those skilled in the art as bark beetles, mainly of the Family Curculionidae. Bark beetles are herbivorous insects of the order Coleoptera with an endophytic habit, which are found in their developmental stages as egg, larva, pupa and / or adult insect, including, for example, Dendroctonus spp., Hylesinus spp., Ips spp. Orthotomicus spp., Pityophthorus spp., Phloeosinus spp, Pseudips spp, Pseudohylesinus spp, Pseudopityophthorus spp. and Scolytus spp, but not limited to certain beetles that cause alteration or damage to coniferous species.

In a preferred embodiment of the invention, bark beetles include, but are not limited to, Dendroctonus adjunctus species, Dendroctonus approximatus, Dendroctonus brevicomis, Dendroctonus frontalis, Dendroctonus jeffreyi, Dendroctonus mexicanus, Dendroctonus rhombectonus, Dendroctonus rhombectonus, Dendroctonus rhombus, Dendroctonus ponytail. Dendroctonus valens, Dendroctonus vitae, Dendroctonus mesoamericanus, Ips bonanseai, Ips calllgraphus, Ips confused, Ips lecontei, Ips pini, Pseudips mexicanus, Pseudohylesinus variegatus, Phloeosinus baumannl, Phloeos, Phloeosinus tacubayae, Pityophthorus, Hylesinus aztecus and Scolytus multistriatus, Scolytus schevyrewi.

In a preferred embodiment of the invention, the forest pests that affect the various species of conifers are shown in Table 1.

Table 1: Bark beetle species of the Curculionidae family affecting species of

Conifers.

In the present invention, entomopathogenic fungi of the genus Beauveria: Beauveria bassiana and / or Beauveria sulfurescens are isolated strains with bioinsecticidal activity, which are useful for the prevention and / or control of homogeneous populations and heterogeneous populations of the forest pest, more particularly bark beetles of the Family Curculionidae. In one embodiment of the invention, the strain of the entomopathogenic fungus of the genus Beauveria is an isolate from a host, preferably the bark beetle in its developmental stages as egg, larva, pupa and / or adult insect, susceptible or involved in the infective process by at least one entomopathogenic fungus of the genus Beauveria, said host can be found under the bark of an infected conifer species. In one embodiment In particular, the strain of the entomopathogenic fungus of the genus Beauveria is an isolate from Dendroctonus mexicanus or Dendroctonus frontalis in its stages of development as an egg, larva, pupa or adult insect, susceptible to a process of mycosis by the entomopathogenic fungus of the genus Beauveria, where the isolated Beauveria bassiana strain is favorable its development in temperate conditions and where the isolated Beauveria sulfureseens strain is favorable its development in temperate-cold conditions with humidity.

In one embodiment of the entomopathogenic fungus of the genus Beauveria of the invention, the entomopathogenic fungus of the genus Beauveria is at least one isolated strain of the genus Beauveria selected from the group of Beauveria bassiana called GIINIF-15 (accession number CM-CNRG TB96) characterized by a mortality of at least 89%, and / or Beauveria sulfureseens called GIINIF-17 (accession number CM-CNRG TB97) characterized by a mortality of at least 96%. The isolated strains referred to in the present invention provide a higher mortality with respect to other isolated strains.

The combination of the isolates of the genus Beauveria provides a synergistic interaction of the isolates of Beauveria bassiana and Beauveria sulfureseens, where the strains produce an infection or a co-infection in compatibility with the biotic conditions of coniferous species in temperate climates and / or cold temperate climates with high humidity, resulting in an efficacy and efficiency of the insecticidal activity for the prevention and control of a homogeneous population or a heterogeneous population of the forest pest of bark beetles of the Curculionidae Family.

These Beauveria strains, the disclosure of which are present in the application, were deposited in the Microorganisms Collection of the National Center for Genetic Resources (CMCNRG), an international depository authority, under the conditions of the Budapest Treaty, for the purposes of the patent procedure. . The strains are registered indicating as person Contact one of the inventors, Dr. Adriana Rosalía Gijón Hernández, as scientific staff of the applicant's CENID-COMEF, acting for and for the applicant the National Institute of Forestry, Agricultural and Livestock Research (INIFAP). The deposition information of the strains is shown in Table 2: Table 2. Deposition information of the Beauveria bassiana strains of the present invention.

The strains belong to the INIFAP Cepart, which were obtained by identifying and isolating strains that show specific activity against the forest pest. In one embodiment of the invention, the process of preparing and obtaining the strains of the entomopathogenic fungus of the genus Beauveria, the process comprises the following stages: a) Identification of coniferous species infected by forest pests. Coniferous species infected by the forest pest known as bark insects (Dendroctonus spp.) Are selected. Once the infected trees were identified, 10x10 cm bark cuts were made from 10 infected and / or damaged trees and transported to the laboratory for morphological analysis. b) Detection of mycosed barking insects. The bark sections are analyzed by microscopy for the detection of mycosed bark beetles, or with a previous mycosis, according to their state or fungal sporulation on the cuticular surface of the bark beetle. The spores or conidia were transferred and seeded in Petri dishes with a culture medium, particularly PDA. c) Isolation of Beauvería bassiana and / or Beauvería sulfurescens strains. Strains of Beauvería bassiana and / or Beauvería sulfurescens are isolated by seed isolation with PDA medium, and incubated at 25 ± 4 ° C, with a relative humidity of 60 ± 10% RH until sporulation of the fungus. Strain isolation methods may comprise other alternative techniques such as serial dilutions or depletion seeding or fractionation of a population of fungi. The concentration of spores or conidia is determined using a Neubauer counting chamber. d) Pathogenicity tests on barking insects. Biological effectiveness tests are carried out on the isolated strains on barking insects in their different stages of development. e) Identification and characterization of the isolated strains. The morphological identification of the structure (conidium, spore, hypha) of the isolated strains, taxonomic keys were made; For the molecular characterization of the strains, a PCR-sequencing of the ITS region is carried out, preferably ITS5 (5'GAAGTAAAAGTCGTAACAAGG-3 ') and ITS4 (5'- TCCTCCGCTTATTGATATGC-3'). These sequences are compared with the species of this type of fungus reported in the Gen Bank (NCB), using BLAST. The isolated strains of the genus Beauvería are found in the conserved regions of the 18S and / or 28S genes respectively. f) Propagation of the isolated strain. The strain is seeded in a PDA medium, with the aim of carrying out spore propagation and obtaining a bbmase from the isolated strain.

In a particular embodiment, the strains of the genus Beauvería comprise a nucleotide sequence, where the Beauvería bassiana strain called GIINIF-15 is shown in SEQ ID N01 corresponding to no. of accession CM-CNRG TB96 and where the Beauvería sulfurescens strain called GIINIF-17 is shown in SEQ ID NO 2 corresponding to no. accession number CM-CNRG TB97.

The Beauvería bassiana strain called GIINIF-15, as used in the present invention, refers to a sequence of 510 base pairs defined by the sequence in the GenBank database with accession number MN658455 that corresponds to SEQ ID N01. The sequence presents 99.80% with different strains of Beauvería bassiana, Beauvería pseudobassiana, Beauvería brongniartii.

The Beauvería sulfurescens strain called GIINIF-17, as used in the present invention, refers to a sequence of 525 base pairs defined by the sequence in the GenBank database with accession number MN658456 that corresponds to SEQ ID NO 2 The sequence shows 99.81% with different strains of Beauvería bassiana.

These isolated strains are of the Beauvería genus, they start the infection process in the bark beetle in any of its stages of development of egg, larva, pupa and / or adult insect, once when they are in contact or the adhesion of the spores or conidia of the isolated strain on the surface of the pest integument. In general, entomopathogenic fungi present the stages of development on the plague: 1) germination of spores, 2) formation of germ tubes and appressoria for fixation in the cuticle; 3) Penetration of germ tubes and colonization of hyphae in internal structures such as muscle tissues, fatty bodies, mitochondria, hemocytes, endoplasmic reticulum and nuclear membrane of the plague, causing mycosis and death of the bark beetle. In this case, the bark beetles die mainly due to the generation of one or more toxins from the entomopathogenic fungus.

In one embodiment of the composition of the present invention, the composition includes at least one entomopathogenic fungus of the genus Beauvería, where the entomopathogenic fungus is selected from the group consisting of Beauvería bassiana (accession no. CM-CNRG TB96) and Beauveria sulfurescens (accession no. CM-CNRG TB97), either alone or in combination thereof. In a more particular embodiment of the composition, the composition comprises a mixture of the strains of Beauvería bassiana GIINIF-15 and Beauvería sulfurescens GIINIF-17, to provide greater protection to the different species of conifers against the harmful effects of a heterogeneous population of forest pests. Said combination of the Beauvería bassiana and Beauvería sulfurescens strains provide higher mortality than the individual strains in the different abiotic factors, due to the adaptability of the entomopathogenic fungus in the different abiotic factors (temperature, humidity and precipitation). In one embodiment of the composition of the present invention, the entomopathogenic Beauvería fungus strains are in the spore or conidium state, an infective unit suitable for their stability and maintenance. The term spore or conidium refers to the sexual and asexual reproductive structure, characteristic of many fungi that allows its survival at temperature or UV rays, and stability under different conditions, as well as its dissemination to a susceptible pest insect.

The compositions of the present invention comprise a concentration between 2.5 X10 ¹⁰ and 3.6 X10 ¹⁰ CFU / mL as a lethal dose of a Beauvería bassiana strain (accession no. CM-CNRG TB96) and / or a concentration between 2.2 X10 ⁸ and 3.4 X10 ⁸ CFU / mL as a lethal dose of a Beauveria sulfurescens strain (accession no. CM-CNRG TB97). The compositions of the present invention are characterized in that they are composed of at least one entomopathogenic fungus; and one or more components comprising a natural substrate, a linear anionic surfactant, a brown seaweed extract, a Moringa oleifera plant extract, and an acceptable carrier, which together allows them to provide the necessary conditions for contact with the fungus. entomopathogen and pest cuticle as well It provides the conditions to carry its development phase and infective process on the plague until its death.

The natural substrate, as used in the present invention, refers to a substance or preparation known to be solid characterized by a content of carbohydrates, vitamins, minerals and amino acids, said natural substrate provides the necessary nutrients for its mycelial growth and to maintain physical conditions such as internal aeration. Any natural substrate known in the state of the art to be suitable for the entomopathogenic fungus can be used in the composition. In this regard, cereals such as wheat husk, rice and corn cob are preferably used. With particular preference, rice of the genus Oryza sativa is used as a solid natural substrate,

The linear anionic surfactant agent as used in the present invention is a surface active agent with the ability to lower surface tension, characterized by an anionic group comprising a linear or branched C ₁₀ to C ₁₄ alkyl chain attached to an anionic group, said anionic surfactant agents have the ability to wet, penetrate and emulsify to improve the contact of the entomopathogenic fungus and the cuticular surface of the forest pest. Any surfactant known in the state of the art to be suitable for wetting and penetrating the particles on the surface of the forest pest can be used in the composition. In this connection, derivatives of the sulfate ion or of sultanates such as sodium dodecyl sulfate and sodium dodecyl bezenesulfonate are preferably used. The soluble extract of brown seaweed as used in the present invention is a preparation or product in solid form, characterized by a content of bioactive substances such as vitamins, minerals, amino acids and wetting agents, said soluble extract of brown seaweed provides components that promote moisture retention. Any soluble extract of seaweed known in the state of the art to be suitable for germination of entomopathogenic fungus, can be used in the composition. In this connection, brown algae of the family Laminariaceae, especially alga gigantea of the genus Macrocystis pyrifera, are preferably used as a soluble extract of brown seaweed.

The Moringa oleifera plant extract as used in the present invention is a preparation or product in liquid form, either in aqueous or ethanolic solution, said Moringa oleifera plant extract is obtained from leaves and / or seeds for the purpose to preserve viability with the entomopathogenic fungus. Any Moringa plant extract known in the state of the art can be suitable for the germination of the entomopathogenic fungus and used in the composition. With particular preference, an aqueous plant extract of Moringa oleifera is used. The acceptable carrier as used in the present invention are powders or granules, said carriers function as transport or vehicle for their protection against adverse factors such as temperature and solar radiation. Any acceptable carrier known in the state of the art to be compatible and suitable for the stability of the entomopathogenic fungus strain can be used in the composition. In this connection, preferably diatomaceous earth, silica and / or combinations thereof are used. With particular preference, diatomaceous earth is used as an acceptable carrier. Diatomaceous earth is an acceptable carrier from fossilized photosynthetic organisms, which allows it to provide minerals and trace elements, and insecticidal activity, also due to its absorption capacity, specific surface and negative electrical charge, which allows it to pierce keratized bodies and absorb components of the waxy structure of the pest insect.

In a preferred embodiment, the biosinsecticidal compositions of the present invention are in powder form. For its preparation, the spores and / or conidia of at least one entomopathogenic fungus of the genus Beauveria are mixed with a natural substrate, a linear anionic surfactant agent, an extract of brown seaweed, a plant extract of Moringa oleifera, and an acceptable carrier. The powder formulation can be dispersed in a wet granulation or suspended in an aqueous solution according to conventional procedures. The solid compositions of the present invention can be in powder, granulate, pellet or tablet form. Additionally, solid forms can be disintegrated or diluted in an aqueous medium for application in liquid forms as solutions or suspensions.

Bioinsecticidal compositions can be stored, packaged, distributed and marketed through commercial packaging without the need for special handling. The compositions can be contained within a packaging material (for example, bags, boxes, vials and any other container because it is suitable for its integrity and stability, it can be suitable for the handling, storage and marketing of the composition. Packaging contains a label or an insert in the packaging material that indicates it can be administered in an amount, with a frequency and duration effective for the prevention and / or biological control of insect pests.

In a particular embodiment, the compositions according to the invention show an improvement in efficacy and efficiency in the prevention and / or control of forest pests, due to the synergistic effect between the entomopathogenic fungus and the components of the natural substrate, a linear anionic surfactant agent. , a brown seaweed extract, a Moringa oleifera extract, and an acceptable carrier which are described in the invention.

The success of germination and penetration not only depends on the type of spore, percentage of germination and susceptibility of the bark beetle, but also on the favorable conditions to favor germination, penetration and aggressiveness of the entomopathogenic fungus, so that the components as a whole provide nutrients, moisture and protection to promote adhesion, germination, penetration and dispersion, as well as the components provide stability against biotic field conditions such as precipitation, temperature or solar rays.

The composition of the present invention can be presented in any suitable form for its administration or application, for example, in solid form such as powders or liquid as suspension, preferably those that make it suitable to be applied according to conventional methods in its handling, either directly or indirectly, or by any other type of application system or equipment.

In a particular embodiment, the application of the compositions comprises the direct application of the composition in at least one segment of the bark where the forest pest is found in its developmental stages as an egg, larva, pupa and / or adult insect. Where the application can be segmented into the parts that comprise the lower segment, the middle segment and the upper segment of the stem, of any of the coniferous species such as pine or cypress. Said compositions can be applied to coniferous species susceptible to attacks by the forest pest, coniferous species with recent attacks and / or coniferous species infected with the forest pest. In this sense, its application to a population of the forest pest guarantees its effectiveness in preventing and / or controlling it. The equipment and procedures for such applications are known in the art and may preferably be those that make it suitable for application in the bark segment of the conifer.

In another optional embodiment of the present invention, the composition can be combined with a chemical agent such as organic insecticide, or another biological agent with insecticidal properties that can offer a synergistic combination for protection to other heterogeneous populations of forest pests, or the generation of long-term sustainable strategies for the preservation, maintenance of coniferous species. The invention is illustrated with preferred embodiments which, however, do not limit the scope of the invention originally described and as defined by the claims in the present application. EXAMPLES

Below, different compositions are presented in a descriptive, non-limiting manner, which are intended to demonstrate the invention:

Example 1: Isolation of strains and morphological characterization of the Beauveria genus isolates. This example illustrates the isolation of the new strains of Beauveria bassiana under accession number CM-CNRG TB96 and Beauveria sulfurescens under accession number CM-CNRG TB97. o Isolated strain of Beauveria bassiana from the temperate forest region of the Sierra Madre Oriental [Sierra Fría: Oak forests 64% of surface area. Temperate climates and at altitudes greater than 800 m].

Three samples of entomopathogenic fungi of Beauveria bassiana present in mycosed bark beetles that were found in 10X10 cm bark samples of pine species trees (Pinus sp) were isolated, the samples were isolated and planted in PDA isolation medium until obtaining of pure strains.

Table 3 shows macrocultural and microcultural morphological characterization data of the isolate of the entomopathogenic fungus of Beauveria bassiana obtained from Dendrodonus mexicanus micosados. (Figure 1a)

Table 3. Morphological characterization of entomopathogenic fungi isolates of Beauveria bassiana obtained from Dendroctonus mexicanas micosados from the Sierra Madre de Aguascalientes.

o Isolated strain of Beauveria sulfurescens from the temperate forest region of the Sierra Madre de Chiapas [Mesophilic forests: Pine-oak forests 61% of surface area. Very humid cold temperate climates and on slopes altitudes above 800 m].

A sample of entomopathogenic Beauveria sulfurescens fungus was isolated in mycosed bark beetles that were found in 10X10 cm bark samples of the trees of pine species (Pinus sp), the samples were isolated and planted in PDA isolation medium until obtaining pure strains. Table 4 shows macrocultural and microcultural morphological characterization data of the entomopathogenic fungus isolate of Beauveria sulfurescens obtained from Dendroctonus mexicanus micosados. (Figure 1b).

Table 4. Morphological characterization of isolates of entomopathogenic fungi of Beauveria sulfurescens obtained from Dendroctonus mexicanus micosados from the Sierra Madre de Chiapas.

Example 2: Evaluation of pathogenicity of strains of the Beauveria genus on the mortality of bark beetles.

This example illustrates the pathogenicity of the new strains of Beauveria bassíana under accession number CM-CNRG TB96 and Beauveria sulfurescens under accession number CM-CNRG TB97. To show the pathogenicity of the Beauveria bassiana isolates, the mortality of bark beetles in their development stages as egg, larva, pupa and / or adult insect was evaluated, the isolated strains were put in contact with the Dendroctonus forest pest. spp. o Isolated strain of Beauveria bassiana from the Sierra Madre Oriental region.

Table 5. Pathogenicity of isolates of entomopathogenic fungi of Beauveria bassiana obtained from Dendroctonus spp. micosados of the Sierra Madre,

For the isolated Beauveria bassiana strain called GIINIF-15, it comprises a lethal dose 50 (LD50) of 2.5 X10 ¹⁰ CFU / mL, and a lethal time 50 (TU,) of 24 hours, or Beauveria sulfurescens isolate from the region of the Sierra Madre de Chiapas.

Table 6. Pathogenicity of isolates of entomopathogenic fungi of Beauveria sulfurescens obtained from Dendroctonus spp. micosados of the Sierra Madre de Chiapas.

For the isolated Beauveria sulfurescens strain called GIINIF-17, it comprises a lethal dose 50 (LD50) of 2.4 X10 ⁸ CFU / mL, and a lethal time 50 (TL50) of 8 hours.

Example 3: Characterization of amino acid sequences of isolated strains of

Beauveria. This example illustrates the identification and phylogenetic relationship of the new strains Beauvería bassiana (CM-CNRG TB96) and Beauvería sulfurescens (CM-CNRG TB97), based on 18S DNA and the ITS sequence obtained.

Taxonomic keys were made; For the molecular characterization of the strains, a PCR-sequencing of the ITS region is carried out, preferably ITS5 (5OGAAGTAAAAGTCGTAACAAGG-3 ') and ITS4 (5'-T CCT CCGCTT ATT GAT ATGC-3'). These sequences are compared with the species of this type of fungus reported in GenBank (NCB), using BLAST. o Beauvería bassiana strain called GIINIF-15 deposited in the Collection of microorganisms of the National Center for Genetic Resources, under the accession number CM-CNRG TB96 from the temperate forest region of the Sierra Madre Oriental.

The sequences obtained were aligned and the percentage of homology and phylogenetic relationship of the new Beauvería bassiana strain was determined, using existing sequences in the NCBI database. The nucleotide sequences were analyzed with the Genbank database, using the BLAST program. The results in Table 7 show the first 10 BLAST results of the 18S DNA and the ITS sequence in the NCBI database respectively.

Table 7. First 10 BLAST results with isolates obtained from an agricultural or forest pest source.

According to the sequence homology of the Beauveria bassiana strain called GIINIF-15, the present strain shows at least 99.80% homology with strains of Beauveria bassiana, Beauveria pseudobassiana or Beauveria brongniartii. However, as shown in figure 2, there are significant differences in their identification compared to the isolated strains of the genus Beauveria obtained from a native pest insect host.

The isolated strain of Beauveria bassiana called GIINIF-15, shows a homology of at least 99.80% with sequences of Beauveria bassiana, Beauveria pseudobassiana, Beauveria brongniartii. o Beauveria sulfurescens strain called GIINIF-17 deposited in the Collection of microorganisms of the National Center for Genetic Resources, under accession number CM-CNRG TB97 from the temperate forest region of the Sierra Madre de Chiapas.

The sequences obtained were aligned and determined percentage of homology and relationship Phylogenetics of the new Beauveria sulfurescens strain, using existing sequences in the NCBI database. The nucleotide sequences were analyzed with the Genbank database, using the BLAST program. The results in Table 8 show the first 10 BLAST results of the 18S DNA and the ITS sequence in the NCBI database respectively.

Table 8. First 10 BLAST results with isolates obtained from an agricultural or forest pest source.

According to the sequence homology of the Beauveria bassiana strain called GIINIF-17, the present strain shows at least 99.81% homology with Beauveria bassiana strains. However, as shown in Figure 3, there are significant differences in its identification in comparison with the isolated strains of the genus Beauvería obtained from a native pest insect host.

The isolated Beauvería sulfurescens strain called GIINIF-17 shows a homology of at least 99.81% with Beauvería bassiana sequences, but by doing the phylogenetic analysis with Bayesian inference it was determined that it belongs to Beauvería sulfurescens.

Example 4: Comparative evaluation of the mortality percentage of the Beauvería isolates in pest insects of the genus Dendroctonus spp. and / or Ips spp.

A comparison was made between the percentages of mortality obtained by Beauvería sulfurescens called GIINIF-17 (accession number CM-CNRG TB97), Beauvería bassiana GIINIF-15 (accession number CM-CNRG TB96) and a control, where a percentage mortality in the insect of the genus Dendroctonus spp. 93.3% for Beauvería sulfurescens and 83.6% for Beauvería bassiana (Fig. 4). And a percentage of mortality in the insect of the genus Ips spp. 90% for Beauvería sulfurescens and 80% for Beauvería bassiana (Fig. 5).

Where a is Beauvería sulfurescens, b Beauvería bassiana and c the control.

Example 5: Compositions with isolates of the Beauvería genus.

Preparation of a composition comprising: a) at least one isolate of an entomopathogenic fungus; b) a natural substrate; c) a linear anionic surfactant; d) a soluble extract of brown seaweed; e) a plant extract of Moringa oleifera; and f) an acceptable carrier. Table 9 shows the different combinations of the qualitative composition.

Table 9. Qualitative composition combinations.

Once the biomass has been mixed with the components, the mixing is carried out according to conventional mixing procedures and the composition is dried at 25 ° C to 32 ° C to obtain a powder or granulate. The formulations represented significant differences in recovery and germination between the isolated strains of entomopathogenic fungi of the Beauvería genus. Since various changes can be made to the aforementioned combinations without departing from the invention, it is intended that all matters contained in the description and the accompanying examples are to be construed as illustrative and not in a limiting sense. Table 10 shows the composition with a concentration range that includes the best recovery and germination performance among the isolated strains of entomopathogenic fungi of the Beauvería genus.

Table 10. Qualitative composition of the present invention.

The compositions can be stored and transported through commercial packaging without the need for special handling. Any container or wrapper being suitable for its stability can be suitable for the handling, storage and distribution of the composition.

Example 6: Evaluation of the effect, time and interaction of the components of the formulation on the mortality of bark beetles.

To test the effect of treatment, time and its interaction on the mortality of the debarkers, the data were compared by means of a factorial ANOVA with the STATISTICA 7 package. Subsequently, for those factors that were statistically significant, Tukey tests were performed (p £ 0.05) .

Statistical analysis recorded a significant effect of time, treatment and time x treatment interaction on the percentage of mortality of bark beetles (P <0.0001). No mortality was recorded in the debarkers with the H + LV and H + D5 treatments. On the other hand, the 8D35 treatment obtained the best results in terms of the percentage of mortality in debarkers (Figure 6 which presented significant differences on mortality at 24 h and 48 h with 47% and 97% respectively, in addition, at 72 h 100% is recorded, while the average mortality percentage for the H + D5 + 103 treatment was 3.33%, which was not statistically significant according to time.

Example 7: Evaluation of the percentage of mortality in cockroaches of the composition of the present invention A test was performed where mortality obtained by a control and the composition of the present invention was determined, which comprises: Beauveria sulfurescens called GIINIF-17 (accession number CM-CNRG TB97) and Beauveria bassíana GIINIF-15 (accession number CM-CNRG TB96), where a percentage of mortality was observed in the insect of the genus Dendroctonus spp. 82% (Fig. 7).

Where a is the composition of the present invention and c the control.

The composition of the present invention comprises the following improvements:

Example 8: Evaluation of the effect, time and interaction of the composition of the present invention with respect to a commercial composition for biological control on the mortality of debarkers.

To test the effect of the treatment, time and its interaction on the mortality of the debarkers, the data were compared by means of a factorial ANOVA with the STATISTICA 7 package. Subsequently, for those factors that were statistically significant, Tukey's tests were performed (p≤ 0.05). Statistical analysis recorded a significant effect of time, treatment, and time * treatment interaction on the percentage of mortality of bark beetles (P <0.0001). No mortality was recorded in the debarkers with the TOC treatment (control). On the other hand, no statistically significant differences were obtained in terms of debarking mortality with the 8DR5 treatment at 24 hrs and the MCB at 48 hrs, with percentages of 43.3% and 53.3% respectively (Figure 8). Likewise, the mortality recorded with the 8DR5 treatment at 48 hrs (93.3%) is statistically similar compared to the MCB treatment at 72 hrs (90%). The aforementioned suggests that with the 8RD5 treatment a mortality percentage similar to the MCB is obtained but in less time, with the MCB a mortality percentage of 90% is recorded at at 72 hrs while with the 8DR5 at 48 hrs 93.3% of mortality was registered and at 72 hrs a 100% was obtained. In addition, the highest mortality in debarking with the MCB treatment was recorded at 72 hours, while, for the 8DR5 treatment, despite the fact that 100% mortality was obtained at 72 hours, the highest mortality was achieved at 48 hours. that statistically, there are no significant differences with the results obtained at 48 hrs and 72 hrs with the 8DR5 treatment.

The composition of the present invention comprises the following improvements: a) It provides a composition that includes a synergistic combination of isolated strains of the Beauveria genus for the prevention and biological control of heterogeneous populations of forest pests of bark beetles of the Curculionidae family, said compositions they are effective and efficient in abiotic factors both in vitro and in situ.

In this way, the characteristics of the present invention provide the following advantages: b) Isolation of bioregulatory agents native to Mexico with the ability to actively infect and cause high mortality to a great diversity of bark beetles of the Curculionidae Family that affect the conifers. c) Obtaining long-term sustainable biological control alternatives for the protection of ends, since they do not require the use of insecticides that produce polluting residues that are harmful to the health of workers in the forestry sector and that are harmful to ecosystems. d) Effective and efficient compositions that can be administered in various environmental conditions, where coniferous species can be susceptible to attack or there is presence of forest pests of bark beetles in their development stages such as egg, larva, pupa and / or insect adult. e) Provides an economical alternative for the timely management of forest pests.

Claims

CLAIMS Having described the invention, the content of the following claims is claimed as property:

1. An isolated strain of the Beauveria genus, characterized in that the strain is the Beauveria bassiana species called GIINIF-15 with deposit in the Collection of microorganisms of the National Center for Genetic Resources (CMCNRG), under the accession number CM-CNRG TB96 or a mutant of it, with insecticidal capacity.

2. The isolated strain of the Beauveria genus according to claim 1, characterized in that it has a DNA sequence corresponding to SEQ ID NO:

1

3. The isolated strain of the Beauveria genus according to claim 2, characterized in that the DNA sequence contains a nucleotide length of 510 base pairs.

4. An isolated strain of the Beauveria genus, characterized in that the strain is the species

Beauveria sulfurescens called GIINIF-17 deposited in the Collection of microorganisms of the National Center for Genetic Resources (CMCNRG), under the accession number CM-CNRG TB97 or a mutant thereof, with insecticidal capacity.

5. The isolated strain of the Beauveria genus according to claim 4, characterized in that it has a DNA sequence corresponding to SEQ ID NO: 2.

6. The isolated strain of the Beauveria genus according to claim 5, characterized in that the one DNA sequence contains a nucleotide length of 525 base pairs.

7. The use of the isolated strains of the Beauveria genus according to claims 1 to 6, for the prevention and / or biological control of pest insects.

8. The use of an isolated strain of the Beauveria genus according to claim 7, wherein the pests are selected from a group comprising Coleoptera, Dermaptera, Diptera, Dictyoptera, Embioptera, Hemiptera, Hymenoptera, Isoptera, Lepidoptera, Orthoptera, Plecoptera , Phithiraptera, Psocoptera, Siphonaptera and Thysanoptera.

9. The use of the isolated strains of the Beauveria genus according to claims 1 to 6, for the preparation of bioinsecticidal compositions.

10. A bioinsecticidal composition comprising: a) Beauveria genus isolate according to claims 1 or 4; b) an acceptable carrier.

The bioinsecticidal composition according to claim 10, wherein the excipients comprise a natural substrate, a diluent, a surfactant agent, a sunscreen agent, a stabilizing agent and an acceptable carrier.

12. The bioinsecticidal composition according to claim 10, wherein the composition is a solid composition in the form of powder, granules, pellets or tablets.

13. The bioinsecticidal composition according to claim 10, wherein the composition is a liquid composition in the form of a solution or suspension.

14. A method for the prevention and / or control of pest insects, characterized in that it consists of putting the pest insect in contact with a strain according to claims 1 or 4.

15. A method for the prevention and / or control of insect pests, according to claim 14, characterized in that the method comprises at least one application directly or indirectly.

16. A synergistic composition characterized in that it comprises the strains of the genus Beauveria: an isolated strain of Beauveria bassiana called GIINIF-15 (accession number CM-CNRG TB96), an isolated strain of Beauveria sulfurescens called GIINIF-17 (accession number CM- CNRG TB97) and an acceptable carrier: for the prevention and / or control of pests.

17. A composition characterized in that it comprises the strains of the genus Beauveria according to claim 9, wherein the pests are selected from a group comprising Coleoptera, Dermaptera, Diptera, Dictyoptera, Embtoptera, Hemiptera, Hymenoptera, Isoptera, Lepidoptera, Orthoptera, Plecoptera , Phithiraptera, Psocoptera, Siphonaptera and Thysanoptera.

18. The bioinsecticidal composition according to claim 10, wherein the composition is a solid composition in the form of powder, granules, pellets or tablets.

19. The bioinsecticidal composition according to claim 10, wherein the composition is a liquid composition in the form of a solution or suspension.

20. A composition for the prevention and / or control of pests, characterized in that the composition comprises: a) at least one isolated strain of an entomopathogenic fungus of the genus Beauveria] b) a natural substrate; c) a linear anionic surfactant; d) a soluble extract of brown seaweed; e) a plant extract of Moringa oleifera; and f) an acceptable carrier; wherein the isolate of an entomopathogenic fungus is selected from the group consisting of: Beauveria bassiana (accession no. CM-CNRG TB96) and Beauveria sulfurescens (accession no. CM-CNRG TB96) and combinations thereof; and wherein the composition has a mortality percentage in pest insects of approximately 100% in a range of 24 to 48 hours.

21. A composition according to claim 21, wherein preferably the entomopathogenic fungus is a mixture of the isolated strains of Beauveria bassiana (accession no. CM-CNRG TB96) and Beauveria sulfurescens (access no. CM-CNRG TB97).

22. A composition characterized in that it comprises the strains of the genus Beauveria according to claim 21, wherein the pests are selected from a group comprising Coleoptera, Dermaptera, Diptera, Dictyoptera, Embioptera, Hemiptera, Hymenoptera, Isoptera, Lepidoptera, Orthoptera, Plecoptera , Phithiraptera, Psocoptera, Siphonaptera and Thysanoptera.

23. A composition according to claim 22, characterized in that the insects of the order Coleoptera comprising at least one selected species from the group of genera Dendroctonus spp., Hylesinus spp., Ips spp., Orthotomicus spp., Pityophthorus spp., Phloeosinus spp., Pseudips spp., Pseudopityophthorus spp., Scolytus spp., and Pseudohyles spp., and Pseudohyles spp.

24. A composition according to claim 21, characterized in that the natural substrate is selected from the group of wheat husks, rice and corn cob,

25. A composition according to claim 25, characterized in that the natural substrate is preferably rice.

26. A composition according to claim 21, characterized in that the linear ammonium surfactant are derivatives of the sulfate ion or of sulfonates selected from the group of sodium dodecyl sulfate, sodium dodecyl bezenesulfonate and combinations thereof.

27. A composition according to claim 21, characterized in that the soluble extract of brown seaweed is preferably a soluble extract of seaweed of the genus Macrocystis pyrifera.

28. A composition according to claim 21, characterized in that the Moringa oleifera plant extract is selected from the group of aqueous extract of Moringa oleifera leaf, ethanolic extract of Moringa oleifera leaf, aqueous extract of Moringa oleifera seed, ethanolic extract Moringa oleifera seed and combinations thereof.

29. A composition according to claim 29, characterized in that the Moringa oleifera plant extract is preferably an aqueous extract of Moringa oleifera leaf.

30. A composition according to claim 21, characterized in that the acceptable carrier comprises at least one acceptable carrier selected from the group of diatomaceous earth, silica and combinations thereof.

31. A composition according to claim 31, characterized in that the acceptable carrier is preferably diatomaceous earth.

32. A composition according to claim 31, characterized in that it is in the form of a powder, granules, tablet or suspension.

33. A composition according to claim 31, characterized in that it can be contained in any container or envelope that maintains its stability.

34. A method for the prevention and / or biological control of pest insects characterized in that it comprises contacting the pest insect with the composition according to claim 21.