河北省二作区马铃薯疮痂链霉菌的分离鉴定及致病性测定

doi:10.7668/hbnxb.20195796

摘要/Abstract

摘要：

为了明确河北省二作区引起马铃薯疮痂病病原种类,对采集自河北省二作区4个地区不同症状类型的19个马铃薯疮痂病病薯进行分离。对分离得到的菌株进行形态学鉴定、PCR分子鉴定和病原菌致病基因textAB、necl和tomA的检测,进一步采用薯片法以及萝卜苗法对含有致病基因菌株的致病力进行验证。最后,通过16S rDNA对所分离的菌株进行种类鉴定并构建系统发育树。结果表明:19个不同症状类型的病薯中共分离得到6株链霉菌。通过16S rDNA聚类分析,将菌株SJZ-1和CL-1鉴定为疮痂链霉菌;将菌株SJZ-2鉴定为花青蓝链霉菌;将菌株CL-2鉴定为白浅灰链霉菌;将菌株SFC鉴定为嗜热一氧化碳链霉菌;将菌株QY鉴定为欧洲疮痂链霉菌。其中,SJZ-1和CL-1含有textAB、necl和tomA 3种致病基因;QY只含有tomA基因;SJZ-2、CL-2和SFC不含有这3种致病基因。薯片法显示,QY和SJZ-1能在薯片上定殖并产生坏死斑,萝卜苗法显示SJZ-1和QY能抑制萝卜苗生长,而SJZ-2、CL-2和SFC无明显致病性。研究初步明确了河北省二作区疮痂病病原菌的种类和组成,为有针对性地采取防控措施提供理论依据。

关键词: 马铃薯疮痂病, 疮痂链霉菌, 致病力

Abstract:

In order to clarify the types of pathogens causing potato common scab in double-cropping cultivation region of Hebei Province,we isolated different types of symptoms tubers which diseased with potato common scab.Morphological identification,PCR molecular identification,and detection of pathogenic genes textAB,necl,and tomA were performed on the isolated strains.The pathogenicity of the strains containing pathogenic genes was further validated using potato chip and radish seedling methods.Finally, the isolated strains were identified at the species level by 16S rDNA sequencing, and a corresponding phylogenetic tree was constructed.The results showed that a total of 6 strains of Streptomyces were isolated from 19 diseased potatoes with different symptom.Through 16S rDNA sequence and amplification analysis,SJZ-1 and CL-1 were identified as S.scabies,SJZ-2 was identified as S.anthocyanicus,CL-2 was identified as S.albogriseolus,SFC was identified as S.thermocarboxydus,QY was identified as S.europaeiscabiei.Strain SJZ-1 and CL-1 contained three pathogenic genes:textAB,necl and tomA.Strain QY only contained the tomA gene.Strain SJZ-2,CL-2 and SFC did not contain any of these three pathogenic genes.The potato chip method showed that QY and SJZ-1 could colonize and produce necrotic spots on small potato chips,the radish seedling method showed that SJZ-1 and QY could inhibit the growth of radish seedlings,while SJZ-2,CL-2,and SFC had no significant pathogenicity in potato chip and radish seedling.This study have preliminary identified the types and composition of potato scab pathogen in double-cropping cultivation region of Hebei Province,and providing a theoretical basis for targeted prevention and potato common scab.

Key words: Potato common scab, Streptomyces app., Pathogenicity

中图分类号:

李怡璠, 韩冬冰, 齐乐丹, 田梦君, 宋雪, 张语涵, 朱杰华, 赵冬梅, 杨志辉. 河北省二作区马铃薯疮痂链霉菌的分离鉴定及致病性测定[J]. 华北农学报, 2025, 40(S1): 270-276. doi: 10.7668/hbnxb.20195796.

LI Yifan, HAN Dongbing, QI Ledan, TIAN Mengjun, SONG Xue, ZHANG Yuhan, ZHU Jiehua, ZHAO Dongmei, YANG Zhihui. Isolation,Identification and Pathogenicity Determination of Potato Common Scab Streptomyces in Double-cropping Cultivation Region of Hebei Province[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(S1): 270-276. doi: 10.7668/hbnxb.20195796.

http://www.hbnxb.net/CN/Y2025/V40/IS1/270

图/表 9

表1 马铃薯疮痂病病薯采集信息

Tab.1 Collection information of diseased potato with potato common scab

采集地点 Collection place	不同症状病薯数量/个 Number of diseased potatoes with different symptoms	病斑类型 Type of lesion	分离菌株数量 Number of isolated strains	菌株编号 Strain number
秦皇岛昌黎县Changli County of Qinhuangdao	2,2,3	凸起状、平状、凹陷状	2	CL-1、CL-2
石家庄正定县Zhengding County of Shijiazhuang	6,1	凹陷状、平状	2	SJZ-1、SJZ-2
保定市清苑区Qingyuan District of Baoding	3	凸起状	1	QY
河北农业大学实验农场	2	平状	1	SFC
Experimental farm of Hebei Agricultural University

表2 供试引物信息

Tab.2 Information of test primers

扩增基因 Amplified gene	引物 Primer	引物序列(5'-3') Primer sequence(5'-3')	片段大小/bp Size of PCR product	退火温度/℃ Annealing temperature
ITS	PA	TTTGATCCTGGCTCAG	1 535	60
	PH	AAGGAGGTGATCCAGCCGCA
textAB	textAB-F	CCACCAGGACCTGCTCTTC	385	53
	textAB-R	TCGAGTGGACCTCACAGATC
necl	necl-F	ATGAGCGCGAACGGAAGCCCCGGA	700	62
	necl-R	GCAGGTCGTCACGAAGGATCG
tomA	tomA-F	GAGGCGTTGGTGGAGTTCTA	392	53
	tomA-R	TTGGGGTTGTACTCCTCGTC

图1 不同地区马铃薯疮痂病症状类型 A.石家庄正定县;B.秦皇岛昌黎县;C.保定市清苑区;D.河北农业大学实验农场。

Fig.1 Symptom types of potato common scab in different regions A.Zhengding County of Shijiazhuang;B.Changli County of Qinhuangdao;C.Qingyuan District of Baoding;D.Experimental farm of Hebei Agricultural University.

图2 不同地区病原菌形态 A.菌株SFJ-1;B.菌株SJZ-2;C.菌株CL-1;D.菌株CL-2;E.菌株SFC;F.菌株QY。

Fig.2 Morphology of pathogens isolated from different regions A.Strain SFJ-1;B.Strain SJZ-2;C.Strain CL-1;D.Strain CL-2;E.Strain SFC;F.Strain QY.

图3 基于16S rDNA序列构建马铃薯疮痂病病菌的系统发育树

Fig.3 Phylogenetic tree of potato scab based on 16S rDNA sequence

图4 致病基因检测结果

Fig.4 Results of pathogenic gene amplification 1.SJZ-1;2.SJZ-2;3.CL-1;4.CL-2;5.SFC;6.QY。

图5 小薯片法验证不同菌株的致病性结果 A.CK;B.菌株QY;C.菌株SJZ-1;D.菌株SFC;E.菌株SJZ-2;F.菌株CL-2。

Fig.5 Small potato slice method to verify the pathogenicity of different strains A.CK;B.Strain QY;C.Strain SJZ-1;D.Strain SFC;E.Strain SJZ-2;F.Strain CL-2.

表3 萝卜苗法验证菌株致病力

Tab.3 Virulence of the strain verified by radish seedling method

菌株编号 Strain number	萝卜苗平均株高/cm Average plant height of radish seedlings	萝卜苗平均根长/cm Average root length of radish seedlings
SJZ-2	8.00±0.612a	9.80±0.570a
CL-2	7.40±0.652a	9.20±0.908ab
SFC	7.20±0.447a	8.90±0.758ab
QY	5.80±0.758b	7.70±0.277b
SJZ-1	5.70±0.570b	5.40±0.652c
CK	7.40±0.418a	9.00±0.707ab

图6 萝卜苗法验证不同菌株致病力

Fig.6 The radish seedling method to verify the pathogenicity of different strains

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