Species and Pathogenicity Identification of Streptomyces Species Causing Potato Common Scab in Ulanqab Area

doi:10.11924/j.issn.1000-6850.casb2024-0170

Abstract

Abstract:

This research aimed to isolate and identify potato common scab pathogens in soil, and clarify the pathogenic genes and pathogenicity. It is of great significance to carry out breeding and comprehensive prevention and control of scab disease. The collected diseased tubers were identified by tissue separation and coating dilution, and the phylogenetic tree was constructed by 16S rDNA measurement. The pathogenic genes were identified by PCR, and the pathogenicity was tested by radish seedling method, small potato chip method and pot inoculation. Four species of pathogenic Streptomyces were isolated and identified as Streptomyces scabies, Streptomyces thermocarboxydus, Streptomyces rubrogriseus and Streptomyces kanamyceticus. All strains were inoculated with ‘Xisen 3’ for pathogenicity test. The test results showed that four Streptomyces could cause tuber disease, and the pathogenic gene combination was txtAB+/tomA+/nec1+; in this study, a total of 4 pathogenic Streptomyces species were identified. Streptomyces thermocarboxydus, Streptomyces rubrogriseus and Streptomyces kanamyceticus were new pathogens of potato common scab.

Key words: potato, common scab, Streptomyces.spp, isolation and identification, pathogenic gene, 16S rDNA measurement, phylogenetic tree, PCR identified

WANG Yue, CAO Chunmei, CHEN Han, WANG Xiaojiao, YU Qianpeng, LI Xueyang, ZHANG Zhikai, HU Baigeng. Species and Pathogenicity Identification of Streptomyces Species Causing Potato Common Scab in Ulanqab Area[J]. Chinese Agricultural Science Bulletin, 2024, 40(32): 151-156.

Figures/Tables 7

References 27

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扩增目标	引物序列	片段大小/bp
16S rDNA	27F：AGAGTTTGATCCTGGCTCAG	1465
16S rDNA	1492R：TACGACTTAACCCCAATCGC	1465
txtAB	txtAB-F：CCACCAGGACCTGCTCTTC	385
txtAB	txtAB-R：TCGAGTGGACCTCACAGATG	385
tomA	tomA-F：GAGGCGTTGGTGGAGTTCTA	392
tomA	tomA-R：TTGGGGTTGTACTCCTCGTC	392
nec1	Nec1-F：ATGAGCGCGAACGGAAGCCCCGGA	700
nec1	Nec1-R：GCAGGTCGTCACGAAGGATCG	700

扩增目标	引物序列	片段大小/bp
16S rDNA	27F：AGAGTTTGATCCTGGCTCAG	1465
16S rDNA	1492R：TACGACTTAACCCCAATCGC	1465
txtAB	txtAB-F：CCACCAGGACCTGCTCTTC	385
txtAB	txtAB-R：TCGAGTGGACCTCACAGATG	385
tomA	tomA-F：GAGGCGTTGGTGGAGTTCTA	392
tomA	tomA-R：TTGGGGTTGTACTCCTCGTC	392
nec1	Nec1-F：ATGAGCGCGAACGGAAGCCCCGGA	700
nec1	Nec1-R：GCAGGTCGTCACGAAGGATCG	700

程序	16S rDNA	txtAB	tomA	nec1
预变性	95℃ 3 min	95℃ 3 min	95℃ 3 min	95℃ 3 min
变性	95℃ 15 s	95℃ 15 s	95℃ 15 s	95℃ 15 s
退火	58℃ 15 s	48℃ 15 s	60℃ 15 s	55℃ 15 s
延伸	72℃ 1.5 min	72℃ 1 min	72℃ 1 min	72℃ 1 min
再延伸	72℃ 5 min	72℃ 5 min	72℃ 5 min	72℃ 5 min
循环数	32	32	32	32

程序	16S rDNA	txtAB	tomA	nec1
预变性	95℃ 3 min	95℃ 3 min	95℃ 3 min	95℃ 3 min
变性	95℃ 15 s	95℃ 15 s	95℃ 15 s	95℃ 15 s
退火	58℃ 15 s	48℃ 15 s	60℃ 15 s	55℃ 15 s
延伸	72℃ 1.5 min	72℃ 1 min	72℃ 1 min	72℃ 1 min
再延伸	72℃ 5 min	72℃ 5 min	72℃ 5 min	72℃ 5 min
循环数	32	32	32	32

菌株	株高/cm	株高抑制率/%	根长/cm	根长抑制率/%	总长/%	总长抑制率/%
NM-3	6.43±1.10 b	9.77	2.03±0.06 e	70.40	8.47±1.04 e	39.52
NM-4	7.20±0.10 a	-0.98	2.60±0.53 de	62.15	9.80±0.52 d	30.00
NM-7	6.80±0.26 a	4.63	3.03±0.25 d	55.85	9.83±0.15 d	29.76
NM-9	2.93±1.01 b	58.86	2.03±0.84 e	70.40	4.97±1.82 f	64.52
L-1	3.33±0.15 b	53.25	0.97±0.15 f	85.93	4.30±0.10 f	69.29
L-2	7.20±0.30 a	-0.98	4.03±0.15 c	41.29	11.23±0.38 c	19.76
L-3	7.10±0.10 a	0.42	5.47±0.51 b	20.43	12.57±0.61 b	10.24
L-4	7.27±0.12 a	-1.92	5.67±0.15 b	17.52	12.93±0.21 ab	7.62
CK	7.13±0.12 a	-	6.87±0.49 a	-	14.00±0.44 a	-