UDC 632.937

CORN ROOT AREA MYСOCENOSIS UNDER THE INFLUENSE OF STRAW WITH DESTRUCTOR FUNGI AS A FERTILIZER

Ye. P. Kopylov, O. V. Skulovatov

Institute of Agricultural Microbiology and Agroindustrial Manufacture, NAAS, Chernihiv, Ukraine; е-mail: [email protected]

Quantitative and qualitative composition of fungi in the rhizosphere soil of corn was studied. It was shown that introduction of straw into the soil resulted in an increase the number of representatives of Fusarium Link and Bipolaris Shoemaker genus, which often include phytopathogenic and phytotoxic fungi, which can affect plants, reducing crop yield and quality. It has been established that application of Chaetomium globosum 377 fungus as destructor of wheat straw leads to the decrease in rhizosphere of corn plants the number of representatives of these genus and an increase in Trichoderma Pers ex Fr. Thus application of C. globosum 377 as destructor of plant residue supports to increase the antagonistic potential of corn rhizosphere soil and protect plants from pathogens.

Key words: corn, wheat straw, mycocenosis, phytopathogenic fungi, Chaetomium globosum.

In order to ensure a stable level of humus in the arable layer of soil, it is necessary to systematically introduce organic fertilizers, including wheat straw, given the high content of carbon in it and the low cost of this agricultural method. The main disadvantages of using straw as a fertilizer is the long process of its mineralization. Another negative factor can be parasitic fungi, which enter the soil together with plant remnants and develop rapidly, using them as a nutrient substrate. Abrupt increase in the proportion of phytopathogenic fungi in the soil can adversely affect the yield of crops to be grown in the future. Thus, fungi of the genus Bipolaris Shoemaker can significantly inhibit the development of plants, causing diseases commonly called helminthosporiosis. Particularly dangerous are representatives of the genus Fusarium Linkex Fr. They can cause not only fusariosis and lead to the loss of 40-50 % of the crop yields, but also to be the producers of mycotoxins that are dangerous for animals [1]. Among the representatives of ascomycetes, Colletotrichum graminicola (Ces.), a causative agent of corn anthracnose, has been described G. W. Wilson. In the vast majority of cases, there is an involvement of the leaves and tissues of the stem, however, there is evidence of the ability of this species to cause damage to the root system [2]. The known pathogenic fungi of plant root zone also include Fusarium 2 graminearum Schwabe, Phaeocytostroma ambiguum (Mont.) Petr., Pyrenochaetater restris (H. N. Hansen) Gorenz, J.C. Walker & Larson, Rhizoctonia solani J.G. Kühn [2-6]. Modern biological methods can largely neutralize these negative effects. Currently, there is a positive experience of using microorganisms both for accelerating the destruction of plant remnants [7] and for protecting crops from fungal diseases [8]. Consequently, the search for microorganisms that can simultaneously accelerate the straw mineralization in the soil and limit the development of phytopathogenic fungi is relevant. It is known that representatives of the genus Chaetomium can exhibit antagonistic activity against a wide range of phytopathogenic fungi [9]. Biofungicidal preparations for the protection of plants have been developed on their basis [10, 11]. For example, Xk-1 Chaetomiumo livaceum Cookeet Ellis can actively suppress soybean fusariosis [12]. We have obtained the newest fungi Chaetomium globosum Kunzeex Fr. 377, which is an active producer of cellulolytic enzymes and can be used as an effective degrader of plant remnants [13]. The objective of this work was to study the mycocenosis of corn root zone and the prospects of using Chaetomium globosum Kunzeex Fr. 377 as a straw destroyer. Materials and methods. Field experiment with Kremin 200 hybrid corn was conducted on the soddy-medium podzolic soil of the experimental field of the Institute of Agricultural Microbiology and Agroindustrial Development of the NAAS (content of humus is 1.2 % (by Tiuryn), mobile nitrogen 5.0-6.0 mg/100 g of soil (by Tiuryn and Kononov), phosphorus – 11-

12 mg/100 g of soil (by Chirikov), potassium – 12-13 mg/100 g of soil (by Kirsanov), pHsol – 6.0). The norm for sowing of corn seeds is 80,000 plants per hectare. The predecessor: black fallow. Scheme of experiment: 1 – control (without straw and fungal spores); 2 – straw introduction; 3 – introduction of straw treated with C. globosum 377 (2 × 105 colony-forming units per 1 g of straw). Straw (8 t/ha) and nitrogen fertilizers at a rate of 15 kg/ha of active ingredient per 1 ton of straw was introduced close to fall plowing. In the spring, mineral fertilizers were introduced in the rate of N80P70K70. Samples of soil for analysis were taken during milk stage of corn. Isolation and identification of fungi from the soil of corn plan rhizosphere were carried out according to generally accepted microbiological methods. Wort agar (concentration of dry matter sis 4-6 %, pH 7.0) was used as a digest medium. Upon identification of micromycetes, V. I. Bilay [14], M. M. Pidoplichko [15, 16] and T. S. Kyrylenko determinants of fungi were used [17]. Field experiments and records were carried out in accordance with the field study techniques, recommended by B. A. Dospiekhov. The data processing was based on the methods of mathematical statistics [18] and Microsoft Excel software [19]. 3

Results and discussion. Study of the quantitative and qualitative composition of fungi in the rhizosphere of corn plants made it possible to determine the competitiveness and survivability of C. globosum Kunzeex Fr. 377 and its influence on phytopathogenic and saprotrophic fungi inhabiting rhizospheric soil (Table 1-2). According to the data obtained, a new strain of C. globosum 377, which exhibits high cellulolytic activity, survived in corn rhizosphere, where its amount was 30 thousand CFUs per 1 g of soil, which was 4.13 % of the total number of fungi. The introduction of straw into the soil resulted in an increase in the number of representatives of the genus Fusarium Linkex Fr. and Bipolaris Shoemaker, among which, phytopathogenic and phytotoxic fungi are common. The application of C. globosum 377 with straw helped to limit the development of Fusarium Linkex Fr. in the rhizosphere of corn plants, the number of which representatives decreased by 5.14 times and amounted to 2.89 % of the total number of fungi. The representatives of the genus Bipolaris Shoemaker were not found in the control variant and in the variant where the destructor fungi was used, but they were represented in significant amount in the variant where the straw only was used. Consequently, the main source of spreading this fungus in the soil after the black fallow is the plant remnants. Comparison of field experiment variants with respect to the most common fungal genera in the soil of rhizosphere of corn plants has shown that the use of straw as a fertilizer provides a reduction in the number of fungi of the genus Trichoderma Persex Fr. by 1.83 times. In the variant using C. globosum 377, the number of representatives of Trichoderma Persex Fr. was the highest and amounted to 66 thousand CFUs per 1 g of soil, indicating an increase in the antagonistic potential of mycocenosis. Consequently, the use of the strain C. globosum 377, together with wheat straw, reduces the risk of corn root diseases caused by phytopathogenic fungi.

4

Table 1. Influence C. globosum 377 on the number of fungi in the rhizospheric soil of corn plants

Number CFU of fungi, th. per 1 g soil ( )

Variants of experiment

ex Fr. ex

in the aggregate the in Acremonium Fr. ex Link Bipolaris Shoemaker Cladosporium Link Chaetomium Kunze Gliocladium Corda Mucor Micheli Fusarium Link Penicillium Link Trichoderma Fr. ex Pers fungi Other Control (without application of straw 435 ± 16 33 ± 3 0 30 ± 3 0 36 ± 5 24 ± 5 27 ± 3 204 ± 13 33 ± 3 48 ± 11 with C. globosum 377) Application of straw 939 ± 39 36 ± 5 39 ± 3 30 ± 3 0 36 ± 5 30 ± 3 108 ± 5 579 ± 26 18 ± 3 63 ± 6 Application of straw 726 ± 19 33 ± 3 0 18 ± 3 30 ± 3 30 ± 3 18 ± 3 21 ± 3 405 ± 13 66 ± 3 105 ± 6 with C. globosum 377

Table 2. Genus content of fungi (% of total number) in rhizospheric soil of corn plants

Number CFU of fungi in % relative to the total number

Variants of experiment

Acremonium Fr. ex Link Bipolaris Shoemaker Cladosporium Link Chaetomium Kunze Gliocladium Corda Mucor Micheli Fusarium Fr. ex Link Penicillium Link Trichoderma Fr. ex Pers fungi Other Control (without application of straw with C. 7,59 0,00 6,90 0,00 8,28 5,52 6,21 46,90 7,59 11,01 globosum 377) Application of straw 3,83 4,15 3,19 0,00 3,83 3,19 11,50 61,66 1,92 6,73 Application of straw with 4,55 0,00 2,48 4,13 4,13 2,48 2,89 55,79 9,09 14,46 C. globosum 377 5

1. Кудаярова Р. Р. Микотоксины. Проблемы и перспективы развития инновационной деятельности в агропромышленном производстве: матер. Всерос. науч.-практич. конф. в рамках ХVII Междунар. cпец. выставки «АгроКомплекс-2007». ― Уфа : Башкирский ГАУ, 2007. ― Ч. 2. ― С. 79. 2. Root infection and systemic colonization of maize by Colletotrichum graminicola / Serenella A. Sukno, Veronica M. Garcia, Brian D. Shaw, Michael R. Thon // Appl Environ Microbiol. ― 2008. ― Vol. 74, № 3. ― P. 823−832. 3. Dorrance evaluation of Fusarium graminearum associated with corn and soybean seed and seedling disease in Ohio / K. D. Broders, P. E. Lipps, P. A. Paul, A. E. Dorrance // Plant Disease. ― 2007. ― Vol. 91, № 9 ― P. 1155−1160. 4. First report of Phaeocytostroma ambiguum causing maize Stalk Rot in Brazil / [F. M. Aguiar, F. E. Lanza, R. V. Costa, D. D. Silva et al.] // Plant Disease. ― 2016. ― Vol. 100, №12. ― P. 2528. 5. Simple and efficient method for detection and identification of Pyrenochaeta terrestris on maize root / J. Levic, S. Stankovic, V. Krnjaja, A. Bocarov-Stancicc // Crop Protection. ― 2012. ― Vol. 38. ― P. 66−71. 6. Kluth C. Maize genotype susceptibility to Rhizoctonia solani and its effect on sugar beet crop rotations / C. Kluth, M. Varrelmann // Crop Protection. ― 2010. ― Vol. 29. ― P. 230−238. 7. Jaybhaye M. M. Role of bioinoculants in the biodegradation of lingo-cellulosic waste (bagasse) / M. M. Jaybhaye, S. A. Bhalerao // Asian Journal of Science and Technology. ― 2016. ― № 4. ― P. 2830−2833. 8. Istifadah N. Endophytic Chaetomium globosum reduces development of tan spot in wheat caused by Pyrenophora tritici-repentis / N. Istifadah, P. A. McGee. // Australasian Plant Pathology. ― 2006. ― № 35. ― P. 411−418. 9. Zhang H. Expressed sequence tags-based identification of genes in the biocontrol agent Chaetomium cupreum / H. Zhang, Q. Yang. // Appl Microbiol Biotechnol. ― 2007. ― №74. ― P. 650−658. 10. Якуба Г. В. Особенности тактики применения микробиологи­ческих препаратов и их эффективность при защите яблони от парши в Краснодарском крае / Якуба Г. В. // Биологическая защита растений – основа стабилизации агроэкосистем : мат. докл. междунар. научно-практич. конф. (29.09-1.10.2004). ― Краснодар, 2004. ― Вып. 3. ― С. 246−248. 11. Надкерничний С. П. Застосування мікробного препарату хетоміка як засобу підвищення урожайності ярого ячменю / С. П. Надкерничний, Є. П. Копилов // Сільськогосподарська мікробіологія. ― 2005. ― № 1. ― С. 30−42. 6

12. Маслиенко Л. В. Штамм гриба Хk-1 Сhaetomium olivaceum Сook et Еllis – продуцент микробиопрепарата для снижения вредоносности фузариоза на сое / Л. В. Маслиенко, Д. А. Курилова // Масличные культуры. ― 2016. ― № 2. ― С. 73−84. 13. Kopilov E. P. Physiological characteristics of mold producing сellulolytic enzymes Chaetomium globosum 377 / E. P. Kopilov, O. V. Skulovatov // Microbiological aspects of optimization of the production process of cultured crops : proceedings of the International Scientific and Practical Internet Conference (Chernihiv, June 16-18, 2015). ― Chernihiv–Nizhyn : Publisher PE Lysenko N.M., 2015. ― P. 12−13. 14. Билай В. И. Фузарии / В. И. Билай ― К. : Наукова думка, 1977. ― 442 с. 15. Пидопличко Н. М. Пенициллии / Н. М. Пидопличко ― К. : Наукова думка, 1972. ― 151 с. 16. Пидопличко Н. М. Грибы-паразиты культурных растений. Определитель / Н. М. Пидопличко ― К. : Наукова думка, 1977. ― Т. 1. ― 295 с. ― Т. 2. ― 300 с. 17. Кириленко Т. С. Определитель почвенных сумчатых грибов / Т. С. Кириленко ― К. : Наукова думка, 1978. ― 264 с. 18. Доспехов Б. А. Методика полевого опыта (с основами статистической обработки результатов исследований) / Б. А. Доспехов. ― М. : Агропромиздат, 1985. ― 351 с. 19. Коросов А. В. Компьютерная обработка биологических данных / А. В. Коросов, В. В. Горбач. ― Петрозаводск : ПетрГУ, 2016. ― 96 с. 7

МИКОЦЕНОЗ КОРНЕВОЙ ЗОНЫ МІКОЦЕНОЗ КОРЕНЕВОЇ ЗОНИ РАСТЕНИЙ КУКУРУЗЫ ПРИ РОСЛИН КУКУРУДЗИ ЗА ИСПОЛЬЗОВАНИИ CHAETOMIUM ВИКОРИСТАННЯ CHAETOMIUM GLOBOSUM КАК ДЕСТРУКТОРА GLOBOSUM ЯК ДЕСТРУКТОРА СОЛОМЫ СОЛОМИ Копылов Е.П., Скуловатов А.В. Копилов Є.П., Скуловатов О.В. Институт сельскохозяйственной Інститут сільськогосподарської микробиологии и агропромышленного мікробіології та агропромислового производства НААН, г. Чернигов виробництва НААН, м. Чернігів

Исследован количественный и Досліджено кількісний і якісний склад качественный состав грибов в ризосферной грибів у ризосферному ґрунті рослин почве растений кукурузы. Показано, что кукурудзи. Показано, що внесення соломи в внесение соломы в почву приводило к росту ґрунт призводило до зростання числа количества представителей родов представників родів Fusarium Link та Fusarium Link и Bipolaris Shoemaker, среди Bipolaris Shoemaker, серед яких часто которых часто встречаются трапляються фітопатогенні та фитопатогенные и фитотоксические фітотоксичні гриби, здатні уражувати грибы, способные поражать растения, рослини, знижуючи урожай та його якість. снижая урожай и его качество. Встановлено, що використання гриба Установлено, что использование гриба Chaetomium globosum 377 як деструктора Chaetomium globosum 377 как деструктора пшеничної соломи сприяє зниженню у пшеничной соломы способствует ризосфері рослин кукурудзи чисельності снижению в ризосфере растений кукурузы представників зазначених родів та численность представителей указанных зростанню кількості Trichoderma Pers ex родов и росту числа Trichoderma Pers ex Fr. Fr. Застосування C. globosum 377 як Применение C. globosum 377 как деструктора рослинних решток дозволяє деструктора растительных остатков підвищити антагоністичний потенціал позволяет повысить антагонистический ризосферного ґрунту кукурудзи та потенциал ризосферной почвы кукурузы и захистити рослини від збудників защитить растения от возбудителей захворювань. заболеваний.

Ключевые слова: кукуруза, пшеничная Ключові слова: кукурудза, пшенична солома, микоценоз, фитопатогенные солома, мікоценоз, фітопатогенні гриби, грибы, Chaetomium globosum. Chaetomium globosum.