doi:10.14720/aas.2019.114.1.11
Original research article / izvirni znanstveni članek
Entomopathogenic fungus, Lecanicillium lecanii R. Z are & W. Gams anchored into MCM-41: A new and effective bio-insecticide against Brevi-coryne brassicae (Linnaeus, 1758) (Hom: Aphididae) to protect cabbages
Asmar SOLEYMANZADE 1 2, Fereshteh KHORRAMI \ Hana BATMANI 3, Khadijeh OJAGHI AGH-BASH 4, Youbert GHOSTA 5
Received June 22, 2018; accepted August 30, 2019. Delo je prispelo 22. junij 2018, sprejeto 30. avgusta 2019.
Entomopathogenic fungus, Lecanicillium lecanii R. Z are & W. Gams anchored into MCM-41: A new and effective bio-insecticide against Brevicoryne brassicae (Linnaeus, 1758) (Hom: Aphididae) to protect cabbages
Abstract: Brevicoryne brassicae is a significant pest of cultivated cabbages and vegetable crops in the world. The present study was carried out to examine a potential strategy to enhance the insecticidal activity of Lecanicillium lecanii for cost-effective management of B. brassicae. The insecticidal efficacy of pure entomopathogenic fungus (PEF) and MCM-41 (Mobil Composition of Matter) L. lecanii were assessed against the cabbage aphid under laboratory and greenhouse conditions. The fungus was supported on MCM-41 and was completely characterized by Scanning Electron Microscope (SEM), thermogravimetric analysis (TGA) and Fourier transform infrared (FT-IR) techniques. LC50 values of PEF and MCM-41@fungus were 1.9x106 and 2.5x104 and 2.0x107 and 2.0x105 conidia/ml on adults of B. brassicae under laboratory and greenhouse conditions, respectively. Bioassays demonstrated that MCM-41@fungus significantly decreased LC50 values of entomopathogenic fungus and it was more toxic than L. lecanii at adult stage of the pest. The results showed that pure L. lecanii and its nano-formulation could play key roles as bio-pesticides in B. brassicae management programs.
Key words: Brevicoryne brassicae; Lecanicillium lecanii; MCM-41@fungus; virulence
Entomopatogena gliva, Lecanicillium lecanii R. Zare & W. Gams, vključena v MCM-41: Novi učinkoviti bio-insekticid za zatiranje mokaste kapusove uši (Brevicoryne brassicae (Linnaeus, 1758) (Hom: Aphididae)) pri zaščiti zelja
Izvleček: Mokasta kapusova uš (Brevicoryne brassicae) je pomemben škodljivec zelja in drugih zelenjadnic širom po svetu. Raziskava je bila izvedena za preučitev potencialne strategije povečanja insekticidne aktivnosti glive Lecanicillium le-canii za učinkovito in poceni zatiranje mokaste kapusove uši. Insekticidna učinkovitost čistega pripravka entomopatogene glive (PEF) in njene vključitve v MCM-41 (Mobil Composition of Matter)@L. lecanii) je bila ocenjena na kapusovi mokasti uši v laboratoriju in v rastlinjaku. Gliva, ki je bila vključena v MCM-41, je bila podrobno opisana z vrstičnim elektronskim mikroskopom (SEM), termogravimetrično analizo (TGA) in Fourierjevo transformacijsko unfrardečo tehniko (FT-IR). LC50 vrednosti za odrasle osebke mokaste kapusove uši so bile za PEF in MCM-41@gliva 1,9 x 106 in 2,5 x 104 ter 2,0 x 107 in 2,0 x 105 konidijev/ml v laboratoriju, oziroma rastlinjaku. Biotest je pokazal, da je kombinacija MCM-41@gliva značilno zmanjšala LC50 vrednosti entomopatogene glive in, da je bila bolj toksična za odrasle uši kot gliva sama. Rezultati so pokazali, da lahko imajo čiste kulture glive L. lecanii in njeni nano pripravki ključno vlogo kot biopesticidi v programih biološkega uravnavanja mokaste kapusove uši.
Ključne besede: Brevicoryne brassicae; Lecanicillium lecanii; MCM-41@gliva; virulenca
1	Islamic Azad University, Urmia Branch, Urmia, Iran
2	Corresponding author, e-mail: soleymanzade.a@gmail.com
3	Islamic Azad University, Sanandaj Branch, Sanandaj, Iran
4	Islamic Azad University, Ajabshir Branch, Ajabshir, Iran
5	Urmia University, Department of Plant Protection, Faculty of Agriculture, Urmia, Iran
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1 INTRODUCTION
Cabbage (Brassica oleracea L. var. capitata) is one of the significant vegetables throughout the world such as Iran. Brassica crops are attacked by many species of insect pests that reduce the quantity and quality of them (Neupane, 1999). Brevicoryne brassicae (Lineus, 1758), (Hom: Aphididae) is one of the serious pests of crucifer crops throughout the world such as Iran (Mousavi An-zabi et al., 2013). The pest causes severe damage by direct feeding or indirect via transfer of viral diseases that could be seriously led to plant destruction (Abdu-Allah, 2012). Heavily infested plants become covered with a mass of aphids that can finally lead to leaf decay and plant death (Griffin & Williamson, 2012). Today, applying chemical insecticides are still considered the foremost and the most important action to manage insect pests. Nevertheless, relying on chemical insecticides has been resulted in adverse effects on environmental and human health. Abuse of non-selective chemical insecticides can destroy the natural enemies and beneficial organisms and induce problems such as development of pest resistance (Sharma & Gupta, 2009). Therefore, these adverse effects supported the development of alternative pest management tactics in which microbial controls may play principle roles (Collantes et al., 1986; Llanderal-Cázares et al., 1996). Entomopathogenic fungi are efficient micro-bial control agents of Homopteran pests (Goettel et al., 2008). Lecanicillium lecanii R. Zare & W. Gams is an entomopathogenic fungus that its mycelium produces a cyclodepsipeptide toxin called bassianolide (Suzuki et al., 1977; Kanaoka et al., 1978). Insect mortality is caused by secrete of mycotoxins and extreme fungal growth (Burg-es, 1981). In spite of the potential of entomopathogenic fungi in the pest control, these bio-control agents have some defects (including sensitivity to environmental factors such as moisture, light, and temperature) that have limited their applications in storage, greenhouse and field conditions. So, these problems will be overcome through recent technological advances such as nanotechnology that will permit future use of entomopathogenic fungi in crop production systems and the nano-formulation approaches can improve their efficacy and pathogenicity. The hexagonal array of uniform mesoporous of MCM-41 with particular properties such as exceptionally high surface areas and high pore volume (Rath & Parida, 2011) attracted our interest in applying it in plant protection and B. brassicae management. Since it has 1D uniform mesopores, the entomopathogenic fungi can be grafted into the MCM-41. Consequently, we selected MCM-41 as a support which belongs to the M41S family that mainly made up of silica, SiO2 (Rath & Parida, 2011; Shy-lesh & Singh, 2005). Silica has unique benefits as a sup-
port like extraordinary thermal and chemical stability, ease of handling, and abundance of exposed silanol (Si-OH) groups (Abdollahi-Alibeik & Pouriayevali, 2012).
2 MATERIALS AND METHODS
2.1	INSECT REARING
Adults of B. brassicae were collected from the cabbage fields in West Azarbaijan Province, Urmia, Iran and then transferred to cabbage plants var. capitata growing on a mixture of soil, sand and manure in plastic plant pots in the greenhouse at 22 ± 2 °C, 70 ± 5 % RH and a photoperiod of 12 L: 12 D. These colonies were rearing on cabbages for 3-4 generations. Cabbages were replaced with new 4-7 leaf stage cabbage plants at every eight days. Trials were carried out with cohorts of apterous adults. Seedlings used for producing leaf disks for entomopathogenic fungus bioassays in Petri dishes.
2.2	MICROORGANISM AND CULTURE MEDIA
The entomopathogenic fungus, Lecanicillium lecanii (IRAN229) was provided from Mycology collections of Urmia University, Urmia, Iran. Fungal species was cultured in potato dextrose agar (PDA) at 25 ± 1 °C. After 2 weeks (enough growth and sporulation of fungus), conidia were scrapped to make an aqueous suspension with 0.02 % Tween-80. The conidia suspension was filtered through three layers of sterile cheesecloth to remove mycelium and was enumerated with an improved Neubauer haemacytometer (Weber Scientific International Ltd., United Kingdom). Before the bioas-say tests, spore viability percentage was determined by inoculating plates of PDA with the suspension. After 24 h, the germination rate was observed under a light microscope. Germination was considered positive when the length of germ tube was as long as the width of the conidia (Hall, 1981). The germination rate of L. lecanii conidia was 97 %.
2.3 NANO STRUCTURE ANALYSIS
The particle morphology was surveyed by a scanning electron microscope (SEM) (Day Petronic Company, Tehran, Iran), using FESEM-TESCAN MIRA3. The thermo gravimetric analysis (TGA) curves were recorded on a Shimadzu DTG-60 instrument (University of Kurdistan, Sanandaj, Iran). Fourier transform infra-
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Entomopathogenic fungus, Lecanicillium lecanii R. Z are & W. Gams anchored into MCM-41: A new and effective ... to protect cabbages
red (FT-IR) spectra were recorded with KBr pellets on 2.6 BIOASSAYS a Nexus 670 FT-IR spectrometer (Medical Sciences of
Urmia University, Urmia, Iran).	2.6.1 Aphid-dipping in the laboratory and green-
house conditions
2.4 PREPARATION OF SILICEOUS MCM-41
Mesoporous Si-MCM-41 was synthesized through Sol-gel tactic according to Cai et al., (2001). The synthesis procedure of Si-MCM-41 was as follow: In a typical procedure, to a solution containing 480 ml deionized water and 3.5 ml NaOH (2M) which was stirred at 80 °C, 1.0 g (2.74 mmol) of surfactant cetyltrimethylammo-nium bromide (CTAB) was added, when the solution became homogeneous, 5 ml of TEOS was slowly added dropwise into the solution, giving rise to a white slurry. The resulting mixture was refluxed for 2h at the same temperature under continuous stirring. The collected product was filtered, washed with deionized water and dried in an oven at 70°C followed by calcination at 550°C for 5 h with a ramp 2°C min-1 to remove the residual surfactants. Finally, we obtained the mesoporous Si-MCM-41.
2.5 PREPARATION OF MCM-41-ENTOMOPATH-OGENIC FUNGUS
In a 100 ml round bottom flask, a mixture of MCM-41-Cl (1 g), entomopathogenic fungus (10 ml of determined concentrations) and Et3N (1.5 ml) in H2O (50 ml) were stirred under room temperature for 24 h. Then, the final product was dried at room temperature (Abdolla-hi-Alibeik & Pouriayevali, 2012).
The fresh leaves used in aphid-dipping under laboratory conditions came from cabbages grown in the research field of Urmia University, Urmia, Iran. For greenhouse trials, cabbages were grown in pots under greenhouse conditions.
To investigate adult sensitivity, each one-day-old aphid was dipped in spore suspensions of pure L. lecanii and MCM-41 @fungus treatments separately under laboratory and greenhouse conditions that determined by preliminary dose setting experiments. The concentration ranges for PEF and MCM-41@fungus in laboratory and greenhouse conditions were 104-108 and 102-106 and 105-109 and 103-107conidia/ml, respectively. For each trial, adults were dipped in the spore suspensions and MCM-41 @fungus treatments separately. For controls, the aphids were immersed in 0.02 % Tween-80 aqueous solution. In the laboratory, when the water had evaporated and the aphids were dry, they were transferred into Petri dishes with fresh cabbage leaves kept at 22 ± 2 °C, 70 ± 5 % RH and a photoperiod of 12 L: 12 D. In the greenhouse, aphids that were immersed in the spore suspensions of PEF and MCM-41 @fungus treatments separately were placed on fresh cabbage leaves. Mortality was recorded after 7 days. There were three replicates of 20 adults per fungal isolate, MCM-41 @fungus and control. In MCM-41@fungus experiments, 0.1 g of each nanocomposite was dispersed in 100 ml distilled water containing 0.02 % Tween-80 until water absorbance was stabilized. After shaking and product dispersion, adult
D1 = 56.82 lit
Figure 1: SEM image of MCM-41@fungus
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aphids were dipped in the corresponding solutions for 10 s.
2.7 DATA ANALYSIS
In order to determine LC50 values, the data were analyzed utilizing the probit procedures with SPSS for Windows® release 24.
3 RESULTS
The Scanning Electron Microscope analysis is an important device for the analysis of the surface morphol-
ogy of a mesoporous structure. The SEM micrograph of MCM-41@fungus mesoporous is shown in Fig. 1. Notably, the particles observe to be in nano range (50-60 nm). The morphology of the mesoporous demonstrates homogeneous, regular, and spherical morphology.
Thermogravimetric analysis (TGA) was employed to select the mass changes of functionalized mesoporous MCM-41.TGA curves for MCM-41 (a) and MCM-41@ fungus (b) are depicted in Fig.2. According to the TGA curve b, an initial mass loss was seen at temperatures below 200 °C because of the removal of physically and chemically adsorbed water molecules inside the pores channels and surface hydroxyl groups. Additionally, at temperatures above 200 °C, large mass losses were occurred which were mainly due to the decomposition of
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Figure 2: TGA thermograms of MCM-41(a) and MCM-41@fungus (b)
Figure 3: FT-IR spectra of MCM-41@fungus (a) and MCM-41 (b) 100 Acta agriculturae Slovenica, 114/1 - 2019
Entomopathogenic fungus, Lecanicillium lecanii R. Z are & W. Gams anchored into MCM-41: A new and effective ... to protect cabbages
Table 1: Toxicity of Lecanicillium lecanii and MCM-41@fungus to one-day-old adults of Brevicoryne brassicae under laboratory and greenhouse conditions
Experimental conditions	Treatments	Slope ± S. E.	x2 (df)	LC50 (conidia/ml) LC90 (conidia/ml)
Laboratory	PEF	3.84 ± 0.04	1.58 (3)	1.9 x 106 (6.5 x 105-6.5 x 106)	2.0 x 1010 (1.4 x 109-3.3 x 1012)
	MCM-41@fungus	4.80 ± 0.05	1.53 (3)	2.5 x 104 (9.3 x 103-8.3 x 104)	1.4 x 108 (1.2 x 107- 1.4 x 1010)
Greenhouse	PEF	3.60 ± 0.03	1.26 (3)	2.0 x 107 (7.1 x 106- 6.6 x 107)	1.5 x 1011 (1.2 x 1010- 1.9 x 1013)
	MCM-41@fungus	4.52 ± 0.05	1.22 (3)	2.0 x 105 (7.8 x 104-6.1 x 105)	8.4 x 108 (8.8 x 107-5.1 x 1010)
PEF: Pure Entomopathogenic Fungus (= non-formulated fungus), 95 % fiducial limit (FL) is shown in parenthesis
covalently bonded organics (200-500 °C) and silanol groups (> 500 °C). This showed that the grafting of fungus had occurred on the inner pore channels of Si-MCM-41.
As shown in Figure 3, curve a, demonstrates stretching vibrations at 1447 cm-1 (C-N), 1729 cm-1(C=O) and a broad band at around 3422 cm-1 (O-H and N-H) which is related to the fungus wall. Curve b, displays the absorption band at 3428 cm-1 is attributed to the stretching vibration of the O-H groups. The bands at 1079, 808 and 456 cm-1 are attributed to the symmetric and asymmetric stretching vibrations of the mesoporous framework (Si-O-Si). By comparing the FT-IR spectrums of MCM-41@fungus (a) and MCM-41 (b) in figure 3, the presence of fungus surrounding the MCM-41 can be clearly acclaimed.
LC50 values of PEF and MCM-41 @fungus were 1.9x106 and 2.5x104 conidia/ml on adults under laboratory conditions and 2.0x107 and 2.0x105 conidia/ml in greenhouse conditions, respectively (Table 1). It is obvious that there is a difference between PEF and MCM-41@fungus, as inferred by the confidence limits of LC50 values (Table 1).
4 DISCUSSION
MCM-41 has attracted great attention because of its large pore size, high thermal stability and extremely high surface area especially above 1000 m2 g-1 (Nikoorazm et al., 2014). The exceptionally high internal surface area, porous structure and uniformity of MCM-41 make it as a cost-efficient host material for a variety of supported catalysts (Nikoorazm et al., 2014). In this study, fungus was grafted into MCM-41 which led to the synthesis of MCM-41@fungus. During the synthesis, MCM-41 was functionalized with the fungus. Functionalization of MCM-41 was performed by the hydroxyl group of en-tomopathogenic fungus. Some studies have revealed
the efficacy of L. lecanii in pest control (Ghaffari et al., 2017; Alavo, 2015; Schreiter et al., 1994; Ramanujam et al., 2017). Our findings confirm L. lecanii pathogenicity against adults of B. brassicae so it will be an efficient candidate for integrated B. brassicae management strategies. According to LC50 values, a significant reduction was seen in the amount of entomopathogenic fungus found in the formulated ones. LC50 values of PEF and MCM-41@fun-gus were 1.9x106 and 2.5x104 conidia/ml on adults of B. brassicae under laboratory and 2.0x107 and 2.0x105 conidia/ml in greenhouse conditions, respectively. Bioas-says demonstrated that nano-formulated L. lecanii significantly decreased LC50 values of fungus and it was more efficient than non-formulated one at adult stage of the pest under laboratory and greenhouse conditions. These results were consistent with the results of Sabbour (2014 b) that studied the efficacy of nano-destruxin of M. an-isopliae (Metchnikoff) Sorokin (against adult females of Hetiracris littoralis Rambur under laboratory conditions. She demonstrated that in nano-formulated samples, LC50 values were significantly decreased. The toxicity bioassays of PEF and MCM-41@fungus indicated that adult stage was more sensitive to nano-formulated L. lecanii than pure entomopathogenic fungus (i.e. MCM-41@fungus exhibited lower LC50 value compared with PEF). Based on our findings, B. brassicae adults were more susceptible to infection by MCM-41@fungus than PEF. Based on our knowledge, this is the first research revealing the susceptibility of B. brassicae adults to MCM-41@entomopatho-genic fungus. The experimental protocol followed in the present study allowed us to provide evidence for MCM-41@entomopathogenic fungus infection.
5 CONCLUSION
Finally, laboratory and greenhouse bioassays have clearly exhibited the pathogenicity of MCM-41@L. leca-
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nii for adults of B. brassicae. These bio-control agents should be considered for the development of a new and environmentally compatible approach for cabbage aphid management in terms of preventing B. brassicae infestations. Further research is required to determine the biological activities of MCM-41@L. lecanii and its persistence after greenhouse and field applications and other factors that may improve its performance against B. bras-sicae throughout its adult stage.
6 REFERENCES
Abdollahi-Alibeik, M., & Pouriayevali, M. (2012). Nanosized MCM-41 supported protic ionic liquid as an efficient novel catalytic system for Friedlander synthesis of qui-nolones. Catalysis Communications, 22, 13-18. https://doi. org/10.1016/j.catcom.2012.02.004 Abdu-Allah, G. (2012). Aphicidal activity of Imidacloprid and Primicarb compared with certain plant extracts on Brevi-coryne brassicae L. and Aphis craccivora Koch. Assiut Journal of Agricultural Science, 43, 104-114. Alavo, T. B. (2015). The insect pathogenic fungus Verticillium lecanii (Zimm.) Viegas and its use for pests control: A review. Journal of Experimental Biology, 3, 337-345. https:// doi.org/10.18006/2015.3(4).337.345 Burges, H.D. (1981). Microbial control of pests and plant diseases
1970-1980. Academic Press,London, 949 pp. Cai, Q., Luo, Z.S., Pang, W.Q., Fan, Y.W., Chen, X.H., & Cui, F.Z. (2001). Dilute solution routes to various controllable morphologies of MCM-41 silica with a basic medium. Chemistry of materials, 13, 258-263. https://doi.org/10.1021/ cm990661z
Collantes, L. G., Raman, K., & Cisneros, F. H. (1986). Effect of six synthetic pyrethroids on two populations of potato tuber moth, Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae), in Peru. Crop Protection, 5, 355-357. https:// doi.org/10.1016/0261-2194(86)90116-X Ghaffari, S., Karimi, J., Kamali, S., & Moghadam, E. M. (2017). Biocontrol of Planococcus citri (Hemiptera: Pseudococ-cidae) by Lecanicillium longisporum and Lecanicillium lecanii under laboratory and greenhouse conditions. Journal of Asia-Pacific Entomology, 20, 605-612. https://doi. org/10.1016/j.aspen.2017.03.019 Goettel, M.S., Koike, M., Kim, J.J., Aiuchi, D., Shinya, R., & Brodeur, J. (2008). Potential of Lecanicillium spp. for management of insects, nematodes and plant diseases. Journal of Invertebrate Pathology, 98, 256-261. https://doi.org/10.1016/j. jip.2008.01.009
Griffin, R.P., & Williamson, J. (2012). Cabbage, Broccoli and other cole crop insect pests HGIC 2203, Home and Garden information center. Clemson cooperative extension, Clem-son University, Clemson, SC. Hall, R. A. (1981). The fungus Verticillium lecanii as a microbial insecticide against aphids and scales. In: Burges HD (ed.) Microbial Control of Pests and Plant Diseases. Academic Press, London, New York. Kanaoka, M., Isogai, A., Murakoshi, S., Ichinoe, M., Suzuki, A.,
& Tamura, S. (1978). Bassianolide, a new insecticidal cy-clodepsipeptide from Beauveria bassiana and Verticillium lecanii. Agricultural and Biological Chemistry, 42, 629-635. https://doi.org/10.1271/bbb1961.42.629 Llanderal-Cazares, C., Lagunes-Tejada, A., Carrillo-Sanchez, J. L., Sosa-Moss, C., Vera-Graziano, J., & Bravo-ojica, H. (1996). Susceptibility of Phthorimaea operculella (Zeller) to insecticides. Journal of Entomological Science, 31, 420-426. https://doi.org/10.18474/0749-8004-31.4.420 MousaviAnzabi, S.H., Nouri-Ghanbalani, G., Eivazi, A., & Ran-ji, A. (2013). Resistance Components of Canola, Brassica napus L. Genotypes to Cabbage Aphid Brevicoryne brassicae (L.). Applied Research in Plant Protection, 2, 85-100. Neupane, F. P. (1999). Field Evaluation of Botanicals for the Management of Cruciferous Vegetable Insect Pests. Nepal Journal of Science and Technology, 1, 77-84. Nikoorazm, M., Ghorbani-Choghamarani, A., Ghorbani, F., Mahdavi, H., & Karamshahi, Z. (2014). Bidentate salen Cu(II) complex functionalized on mesoporous MCM-41 as novel nano catalyst for the oxidative coupling of thiols into disulfides using urea hydrogen peroxide (UHP). Journal of Porous Materials https://doi.org/10.1007/s10934-014-9892-6
Ramanujam, B., Krishna, J., & Poornesha, B. (2017). Field evaluation of entomopathogenic fungi against cabbage aphid, Brevicoryne brassicae (L.) and their effect on coccinellid predator, Coccinella septempunctata (Linnaeus). Journal of Biological Control, 31,168-171. https://doi.org/10.18311/ jbc/2017/16350
Rath, D., & Parida, K. M. (2011). Copper and nickel modified MCM-41 an efficient catalyst for hydrodehalogenation of chlorobenzene at room temperature. Industrial and Engineering Chemistry Research, 50, 2839-2849. https://doi. org/10.1021/ie101314f Sabbour M. M. (2014). Evaluating toxicity of extracted nano-Destruxin against the desert locust Schistocerc agregaria in Egypt. The Journal of Egyptian Academic Society Environmental Development, 15, 9-17. Schreiter, G., Butt, T.M., Beckett, A., Vestergaard, S., & Moritz, G. (1994). Invasion and development of Verticillium lecanii in the western flower thrips, Frankliniella occidentalis. My-cological Research, 98, 1025-1034. https://doi.org/10.1016/ S0953-7562(09)80429-2 Sharma, A., & Gupta, R. (2009). Biological activity of some plant extracts against Pieris brassicae (Linn.). Journal of Biopesticides, 2 26-31. Shylesh, S., & Singh, A. P. (2005).Vanadium-containing ordered mesoporous silicates: Does the silica source really affect the catalytic activity, structural stability, and nature of vanadium sites in V-MCM-41? Journal of Catalysis, 233, 359-371. https://doi.org/10.1016/j.jcat.2005.05.001 Suzuki, A., Kanaoka, M., Isogai, A., Tamura, S., Murakoshi, S., & Ichinoe, M. (1977). Bassianolide, a new insecticidal cyclodepsipeptide from Beauveria bassiana and Verticillium lecanii. Tetrahedron Letters, 18, 2167-2170. https://doi. org/10.1016/S0040-4039(01)83709-6
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