Identification of Trichoderma harzianum CCTH-2 and Its Biological Characteristics,a Biocontrol Fungi Against Corn Stalk Rot

doi:10.7668/hbnxb.20192851

Abstract

Abstract:

Corn stalk rot is harmful to yield and quality of corn,so effective control of corn stalk rot should be carried out as soon as possible.For Fusarium graminearum,the dominant strains of corn stalk rot in Jilin Province,and fourteen Trichoderma sp.strains were isolated from corn rhizosphere soil were subjected to plate confrontation experiment respectively.By observing the inhibitory effect of Trichoderma sp.strains on Fusarium graminearum,the strains of CCTH-2 and CCTH-6 with better inhibitory effect could be screed.The inhibition of Fusarium graminearum by fermentation broth,volatile metabolites and non-volatile metabolites of these two Trichoderma sp.strains were screened again,and inhibitory effect of these two Trichoderma sp.strains on other pathogens.Finally,Trichoderma sp.strain CCTH-2 with good control effect was screened by comprehensive analysis.Morphological and molecular identification showed Trichoderma harzianum,and its biological characteristics were preliminarily studied.The results showed that the inhibition rate of CCTH-2 against Fusarium graminearum was 82.83% under culture conditions of slab face-off method and the fermentation broth,volatile metabolites and non-volatile metabolites of CCTH-2 against Fusarium graminearum was 36.56%,46.75%,32.48%,showed significant inhibition effect.The biological characteristics test showed that PDA was the optimal medium for the growth and sporulation of CCTH-2 colonies,the optimal temperature was 25 ℃,the optimal illunination condition was total illumination,exogenous addition of Fe²⁺ was beneficial to the growth and development of CCTH-2,and CCTH-2 had the ability to adapt to the changes of pH value and salt contene.Therefore,it could be confirmed that Trichoderma harzianum CCTH-2 was a biocontrol bacterium with good control effect and great potential,providing information guarantee and theoretical basis for the subsequent development and utilization of biocontrol bacteria of corn stalk rot.

Key words: Corn stalk rot, Fusarium graminearum, Trichoderma harzianum CCTH-2, Biological characteristics

TONG Yunzheng, YU Huilin, PAN Hongyu, WANG Yingchun. Identification of Trichoderma harzianum CCTH-2 and Its Biological Characteristics,a Biocontrol Fungi Against Corn Stalk Rot[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(S1): 309-317. doi: 10.7668/hbnxb.20192851.

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Figures/Tables 19

Tab.1 Inhibitory rate of Trichoderma spp. to Fusarium graminearum

菌株 Strains	禾谷镰孢 F. graminearum
菌株 Strains	菌落半径/cm The radius of colony	抑制率/% Inhibation rate
CCTH-1	1.10	72.58±0.60bc
CCTH-2	0.69	82.83±0.25a
CCTH-3	1.24	68.92±0.49c
CCTH-4	1.36	66.08±0.56d
CCTH-5	1.09	72.75±0.31bc
CCTH-6	0.70	82.42±0.25a
CCTH-7	0.95	76.33±0.30b
CCTH-8	1.23	69.33±0.18c
CCTH-9	1.21	69.67±0.25c
CCTH-10	1.11	71.83±0.25bc
CCTH-11	1.11	72.33±0.64bc
CCTH-12	1.14	71.42±0.27bc
CCTH-13	1.16	71.00±0.42c
CCTH-14	1.07	73.83±0.45b
CK	4.00	-

Fig.1 Colony growth morphology diagram of Fusarium graminearum(Both sides of culture medium) A.Inhibition activities of fermentation broth of Trichoderma sp. CCTH-2 and CCTH-6 species against Fusarium graminearum(3 d);.B.Inhibition activities of volatile metabolites of Trichoderma sp. CCTH-2 and CCTH-6 species against Fusarium graminearum(5 d);C.Inhibition activities of non-volatile metabolites of Trichoderma sp. CCTH-2 and CCTH-6 species against Fusarium graminearum(36 h).

Tab.2 Inhibitory activities of Trichoderma sp. CCTH-2 and CCTH-6 species against Fusarium graminearum

菌株 Strains	发酵液 Fermentation broth		易挥发性代谢产物 Volatile metabolites		难挥发性代谢产物 Non-volatile metabolites
菌株 Strains	菌落半径/cm The radius of colony	抑制率/% Inhibition rate	菌落半径/cm The radius of colony	抑制率/% Inhibition rate	菌落半径/cm The radius of colony	抑制率/% Inhibition rate
CCTH-2	2.03	36.56±1.21a	2.13	46.75±0.18a	1.06	32.48±1.59b
CCTH-6	2.18	31.87±0.92b	2.28	43.00±0.68b	0.91	42.04±1.59a
CK	3.20	-	4.00	-	1.57	-

Tab.3 Inhibitory activities of Trichoderma sp.CCTH-2 and CCTH-6 species against other pathogenic fungi

菌株 Strains	对照组 Control group	木霉菌CCTH-2 Trichoderma sp. CCTH-2		木霉菌CCTH-6 Trichoderma sp. CCTH-6
菌株 Strains	菌落半径/cm The radius of colony	菌落半径/cm The radius of colony	抑制率/% Inhibition rate	菌落半径/cm The radius of colony	抑制率/% Inhibition rate
立枯丝核菌R.solani	7.50	2.27	69.73±0.86ab	2.63	64.93±0.77b
玉米大斑病菌S. turcica	3.45	1.05	69.57±0.80b	1.11	67.83±0.55b
玉米小斑病菌C. heterostrophus	3.42	1.17	65.79±1.51b	1.08	68.42±1.90b
核盘菌S. sclerotiorum	7.50	2.83	62.27±1.35b	2.57	65.73±1.78b
玉米灰斑病菌C. zeae-maydis	7.50	1.40	81.33±2.26a	1.46	80.53±0.93a
灰葡萄孢B.cinerea	6.07	1.31	78.42±1.87a	1.36	77.59±1.32a

Fig.2 Inhibitory activities of Trichoderma sp.CCTH-2 and CCTH-6 species against other pathogenic fungi

Fig.3 Trichoderma harzianum morphologic characteristic A.Colony on PDA(initial stage);B.Colony reverse on PDA(late stage);C.Conidiophore;D.Conidium.

Fig.4 Phylogenetic tree of Trichoderma harzianum CCTH-2

Fig.5 Colony morphology of Trichoderma harzianum CCTH-2 on different media

Fig.6 Effects of different media on mycelial growth and sporulation of Trichoderma harzianum CCTH-2

Fig.7 Colony morphology of Trichoderma harzianum CCTH-2 under different temperature conditions

Fig.8 Effects of different temperatures on mycelial growth and sporulation of Trichoderma harzianum CCTH-2

Fig.9 Colony morphology of Trichoderma harzianum CCTH-2 under different light types

Fig.10 Effects of different light types on mycelial growth and sporulation of Trichoderma harzianum CCTH-2

Fig.11 Colony morphology of Trichoderma harzianum CCTH-2 under different trace elements

Fig.12 Effects of different trace elements on mycelial growth and sporulation of Trichoderma harzianum CCTH-2

Fig.13 Colony morphology of Trichoderma harzianum CCTH-2 under different pH conditions

Fig.14 Effects of different pH values on mycelial growth and sporulation of Trichoderma harzianum CCTH-2

Fig.15 Colony morphology of T.harzianum CCTH-2 under different salt content conditions

Fig.16 Effects of different salt content on mycelial growth and sporulation of Trichoderma harzianum CCTH-2

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