Identification, Biocontrol and Plant Growth-promoting Potential of Endophytic Bacterial Strain JY-3-1R from Aconitum carmichaelii Debx.

doi:10.13560/j.cnki.biotech.bull.1985.2023-0590

Abstract

Abstract:

In order to provide promising microbial resource for the control of soil-borne diseases in Aconitum carmichaelii Debx., endophytic bacterial strains with biocontrol and plant growth promoting potential were isolated and screened from healthy A. carmichaelii plants. Pure culture method was used to isolate endophytic bacteria, and dual-culture method was to check the antagonistic activity of candidate strains against pathogens. Multi-locus housekeeping gene sequence analysis was to investigate phylogeny of the strain. Enzyme- and siderophore-producing capacities of bacterial strain were measured on plates. Salkowski method was applied to measure indoleacetic acid（IAA）-producing ability of the strain. Field experiment was employed to demonstrate plant growth promoting ability and biocontrol potential against southern blight in A. carmichaelii. One strain JY-3-1R screened from 111 A. carmichaelii endophytic bacteria showed significant antagonistic activity against Sclerotium rolfsii and Fusarium oxysporum in vitro. The JY-3-1R was identified as Bacillus amyloliquefaciens. Cell-free culture filtrate of JY-3-1R presented 100% inhibition on the hyphal growth and sclerotia germination of S. rolfsii. JY-3-1R was able to produce IAA, siderophore, protease, cellulase, glucanase, also had the functional genes of synthesizing iturin, fengyin, surfactin, bacillomycin, bacilysin, bacillaene, and macrolactin. Under field conditions, JY-3-1R inoculation reduced around 40% of southern blight occurrence, whose biocontrol rates ranged from 61.53% to 84.61%. This biocontrol efficiency lasted at least for 30 d. Meanwhile, JY-3-1R inoculation promoted 34.34%, 82.59% and 56.08% of stem, main root and lateral root dry weight, respectively of A. carmichaelii. Altogether, JY-3-1R demonstrated both plant growth promoting and biocontrol potential, and could be used as good candidate of biofertilizer and biocontrol agent of A. carmichaelii.

Key words: endophytic bacteria, biocontrol, plant growth promotion, southern blight

ZOU Lan, WANG Qian, LI Mu-yi, YE Kun-hao, HUANG Jing. Identification, Biocontrol and Plant Growth-promoting Potential of Endophytic Bacterial Strain JY-3-1R from Aconitum carmichaelii Debx.[J]. Biotechnology Bulletin, 2023, 39(10): 246-255.

Figures/Tables 9

References 31

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基因Gene	引物Primer（5'-3'）	反应程序Reaction procedure
16S rRNA	27F: AGAGTTTGATCCTGGCTCAG 1492R: TACGGCTACCTTGTTACGACTT	94℃ for 3 min, 30 cycles of 94℃ for 1 min, 56℃ for 1 min, 72℃ for 2 min, final extension for 72℃ 10 min
atpD	atpDF: RTAATYGGMGCSGTRGTNGAYGT’ atpDR:TCATCCGCMGGWACRTAWAYNGC’	98℃ for 2 min, 35 cycles of 98℃ for 30 s, 56℃ for 1 min, 72℃ for 45 s, final extension for 72℃ 10 min
gyrA	gyrA2F: ATGAGCGATCTGGCCAGAGA gyrA9R: CGCGCCTTGTTCACCTGATA	98℃ for 2 min, 35 cycles of 98℃ for 30 s, 57℃ for 1 min, 72℃ for 45 s, final extension for 72℃ 10 min
rpoB	rpoBF: AGGTCAACTAGTTCAGTATGGAC rpoBR: AAGAACCGTAACCGGCAACTT	94℃ for 3 min, 30 cycles of 94℃ for 30 s, 54℃ for 45 s, 72℃ for 30 min, final extension for 72℃ 10 min

基因Gene	引物Primer（5'-3'）	反应程序Reaction procedure
16S rRNA	27F: AGAGTTTGATCCTGGCTCAG 1492R: TACGGCTACCTTGTTACGACTT	94℃ for 3 min, 30 cycles of 94℃ for 1 min, 56℃ for 1 min, 72℃ for 2 min, final extension for 72℃ 10 min
atpD	atpDF: RTAATYGGMGCSGTRGTNGAYGT’ atpDR:TCATCCGCMGGWACRTAWAYNGC’	98℃ for 2 min, 35 cycles of 98℃ for 30 s, 56℃ for 1 min, 72℃ for 45 s, final extension for 72℃ 10 min
gyrA	gyrA2F: ATGAGCGATCTGGCCAGAGA gyrA9R: CGCGCCTTGTTCACCTGATA	98℃ for 2 min, 35 cycles of 98℃ for 30 s, 57℃ for 1 min, 72℃ for 45 s, final extension for 72℃ 10 min
rpoB	rpoBF: AGGTCAACTAGTTCAGTATGGAC rpoBR: AAGAACCGTAACCGGCAACTT	94℃ for 3 min, 30 cycles of 94℃ for 30 s, 54℃ for 45 s, 72℃ for 30 min, final extension for 72℃ 10 min

菌株 Strain	PDA平板抑菌率 Inhibition rate on PDA plate /%					乌头切片抑菌率 Inhibition rate on A. carmichaelii root slice/%
菌株 Strain	S. rolfsii-1	S. rolfsii-2	F. oxysporum-1	F. oxysporum-2		S. rolfsii-1	S. rolfsii-2	F. oxysporum-1	F. oxysporum-2
JY-3-1 R	53.13±0.39	51.30±1.09	52.07±0.21	44.58±0.18		16.32±9.76	49.68±3.05	42.19±0.09	46.30±3.27

菌株 Strain	PDA平板抑菌率 Inhibition rate on PDA plate /%					乌头切片抑菌率 Inhibition rate on A. carmichaelii root slice/%
菌株 Strain	S. rolfsii-1	S. rolfsii-2	F. oxysporum-1	F. oxysporum-2		S. rolfsii-1	S. rolfsii-2	F. oxysporum-1	F. oxysporum-2
JY-3-1 R	53.13±0.39	51.30±1.09	52.07±0.21	44.58±0.18		16.32±9.76	49.68±3.05	42.19±0.09	46.30±3.27

测定指标 Character/（g·plant^-1）	处理 Treatment
测定指标 Character/（g·plant^-1）	JY-3-1R	CK
茎鲜重 Stem fresh weight	37.68±2.91 a	27.98±3.25 b
茎干重Stem dry weight	9.35±0.70 a	6.96±0.90 b
乌头鲜重Main root fresh weight	28.43±2.43 a	19.44±1.02 b
乌头干重Main root dry weight	5.77±0.78 a	3.16±0.11 b
子根鲜重Lateral root fresh weight	77.17±7.80 a	48.54±5.95 b
子根干重Lateral root dry weight	20.01±1.94 a	12.82±1.91 a