Biocontrol efficacy and action mechanism of Trichoderma harzianum LTR-2 and Arthrobacter ureafaciens DnL1-1 against crown rot of wheat

doi:10.3969/j.issn.1004-1524.2023.06.16

Acta Agriculturae Zhejiangensis ›› 2023, Vol. 35 ›› Issue (6): 1385-1395.DOI: 10.3969/j.issn.1004-1524.2023.06.16

• Plant Protection • Previous Articles Next Articles

Biocontrol efficacy and action mechanism of Trichoderma harzianum LTR-2 and Arthrobacter ureafaciens DnL1-1 against crown rot of wheat

YANG Kai(), CHEN Kai, LI Hongmei, ZHAO Zhongjuan, HU Jindong, LI Jishun^*(), YANG Hetong

Shandong Provincial Key Laboratory of Applied Microbiology, Ecology Institute, Qilu University of Technology (Shandong Academy of Sciences), Jinan 250103, China

Received:2022-07-18 Online:2023-06-25 Published:2023-07-04

Abstract

Abstract:

To assess the synergism control of Trichoderma harzianum LTR-2 and Arthrobacter ureafaciens DnL1-1 against wheat crown rot. The antimicrobial activities of LTR-2 and DnL1-1 against pathogenic fungus Fusarium pseudograminearum were examined using confrontation culture experiment and disease-control pot experiment. Then the inhibitory mechanism of LTR-2 and DnL1-1 against F. pseudograminearum was studied by measuring mycelial growth and spore germination. The results showed that with the decrease of F. pseudograminearum inoculation concentration, the disease index of wheat crown rot gradually decreased, and the control effects of various treatments on wheat crown rot were significantly (P<0.05) different. Among these treatments, the combined inoculation of LTR-2 and DnL1-1 showed the best control effects on wheat crown rot with the values of 34.7%, 50.3% and 87.2%, respectively. In plate confrontation test, the inhibitory rates of LTR-2+DnL1-1 and LTR-2 alone against F. pseudograminearum were both 100%, and there was no significant difference between the two treatments. The microscopic results of confrontation test showed that the morphology of F. pseudograminearum mycelia changed significantly with protoplast condensation, septum increase and mycelium fracture under the effect of LTR-2 alone and LTR-2 and DnL1-1 synergy. The fermentation filtrates of the LTR-2 alone culture and combined culture of LTR-2 and DnL1-1 both had a significant effect on the mycelia morphology of F. pseudograminearum, resulting in mycelial expansion and distortion, and affected the sporulation type, so that only produced conidia, but almost no chlamydospore. In addition, the germination of conidia was inhibited significantly. In conclusion, DnL1-1 had a synergistic effect on the control effect of LTR-2 against wheat crown rot, and the above results afforded scientific basis for the combined application of the two strains.

Key words: Trichoderma harzianum, Arthrobacter ureafaciens, wheat crown rot, biological control

CLC Number:

S435.121

YANG Kai, CHEN Kai, LI Hongmei, ZHAO Zhongjuan, HU Jindong, LI Jishun, YANG Hetong. Biocontrol efficacy and action mechanism of Trichoderma harzianum LTR-2 and Arthrobacter ureafaciens DnL1-1 against crown rot of wheat[J]. Acta Agriculturae Zhejiangensis, 2023, 35(6): 1385-1395.

Figures/Tables 9

Table 1 Effect of LTR-2 and DnL1-1 on disease index of wheat crown rot

处理 Treatment	病情指数Disease index
处理 Treatment	100倍稀释液 Dilution ratio 100×	50倍稀释液 Dilution ratio 50×	20倍稀释液 Dilution ratio 20×
CK	25.8±6.3 a	62.0±7.5 a	79.2±5.2 a
酷拉斯 Kuras	3.3±1.4 c	27.5±7.5 b	51.7±1.4 b
LTR-2+DnL1-1	3.3±3.8 c	30.8±8.8 b	51.7±1.4 b
LTR-2	15.8±6.3 b	42.5±2.5 ab	53.3±7.6 b
DnL1-1	27.5±5.0 a	56.7±11.8 a	71.7±8.0 a

Fig.1 Control effect of LTR-2 and DnL1-1 on wheat crown rot Data on the bars marked without the same lowercase letter indicated significant differences at P<0.05.

Table 2 Effects of different treatments on wheat growth under 20 times dilution inoculation of Fusarium pseudograminearum

处理 Treatment	株高 Plant height/cm	苗鲜重 Fresh weight of seedling/g	根鲜重 Fresh weight of root/g	根冠比 Root shoot ratio/%
CK	23.28±0.81 ab	4.14±0.13 b	1.81±0.30 b	0.44±0.09 b
酷拉斯Kuras	23.84±1.16 ab	5.33±0.56 a	2.78±0.73 a	0.53±0.17 a
LTR-2+DnL1-1	24.42±0.76 a	4.32±0.22 b	2.89±0.30 a	0.67±0.08 a
LTR-2	23.29±0.70 ab	4.54±0.18 b	2.93±0.25 a	0.65±0.07 a
DnL1-1	22.85±1.12 bc	4.14±0.44 b	2.41±0.93 a	0.57±0.19 a

Fig.2 Inhibitory effect of LTR-2 and DnL1-1 on Fusarium pseudograminearum A-D represent the control, DnL1-1, LTR-2 and DnL1-1+LTR-2 confront cultured with Fusarium pseudograminearum for 7 days, respectively.

Fig.3 Effects of LTR-2 and DnL1-1 on mycelial growth of Fusarium pseudograminearum A, Control; B, DnL1-1 confront cultured with Fusarium pseudograminearum; C and D, LTR-2 confront cultured with Fusarium pseudograminearum; E, F and G, DnL1-1+LTR-2 confront cultured with Fusarium pseudograminearum. The black arrows in the figure represented that Trichoderma parasitize Fusarium pseudograminearum in ways of twining, penetrating and clinging, and lead to Fusarium pseudograminearum mycelium protoplasm shrinkage, septum increased, cell became shorter or mycelium broke.

Fig.4 Inhibition of different fermentation filtrates on growth of Fusarium pseudograminearum mycelia A and B represent the results of cultured for 2 and 5 days, respectively. From left to right are the sterile water control group, DnL1-1 fermentation filtrate treatment group, LTR-2 fermentation filtrate treatment group and DnL1-1+LTR-2 combined fermentation filtrate treatment group. The top is the front side, and the bottom is the back side.

Fig.5 Effects of LTR-2 and DnL1-1 fermentation filtrate on mycelia growth of Fusarium pseudograminearum in liquid culture A, B, C and D represent the sterile water, DnL1-1 fermentation filtrate, LTR-2 fermentation filtrate and DnL1-1+LTR-2 fermentation filtrate cultured with Fusarium pseudograminearum, respectively. The black arrows in the figure refer to the area where the hyphae expand, twist or fuse.

Fig.6 Effect of LTR-2 and DnL1-1 fermentation filtrate on sporulation of Fusarium pseudograminearum in liquid culture A, B, C, D and E represent the sterile water, DnL1-1 fermentation filtrate, LTR-2 fermentation filtrate and DnL1-1+LTR-2 fermentation filtrate cultured with Fusarium pseudograminearum, respectively. The black arrows in the figure indicate different spore types, including chlamydospores and conidia.

Fig.7 Effects of LTR-2 and DnL1-1 fermentation filtrate on conidial germination of Fusarium pseudograminearum

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Biocontrol efficacy and action mechanism of Trichoderma harzianum LTR-2 and Arthrobacter ureafaciens DnL1-1 against crown rot of wheat

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