猕猴桃溃疡病病菌的多样性及其检测技术

doi:10.16178/j.issn.0528-9017.20213218

摘要/Abstract

摘要：

猕猴桃细菌性溃疡病是全球重要的植物检疫性病害，丁香假单胞杆菌猕猴桃致病变种被认为是引起猕猴桃细菌性溃疡病的主要病原菌，准确、快速、灵敏的检测方法对预防其扩散和传播至关重要。本文结合国内外相关研究报道，就猕猴桃细菌性溃疡病菌的多样性及其检测技术进行了系统的综述，重点分析比较了现有不同检测技术的优缺点，展望了猕猴桃细菌性溃疡病菌检测技术未来的发展方向，为准确、快速、灵敏地检测猕猴桃细菌性溃疡病菌提供了基础。

关键词: 猕猴桃细菌性溃疡病, 遗传多样性, 检测技术, 丁香假单胞杆菌

中图分类号:

S41-30

胡施祺, 戴德江, 罗金燕, 朱洁, 安千里, 李斌. 猕猴桃溃疡病病菌的多样性及其检测技术[J]. 浙江农业科学, 2022, 63(11): 2652-2657.

图/表 2

参考文献 49

[1]	OPGENORTH D C. Pseudomonas canker of kiwifruit[J]. Plant Disease, 1983, 67(11): 1283.
[2]	SERIZAWA S, ICHIKAWA T, TAKIKAWA Y, et al. Occurrence of bacterial canker of kiwifruit in Japan: Description of symptoms, isolation of the pathogen and screening of bactericides[J]. Japanese Journal of Phytopathology, 1989, 55(4): 427-436.
[3]	TAKIKAWA Y, SERIZAWA S, ICHIKAWA T, et al. Pseudomonas syringae pv. actinidiae pv. nov.: The causal bacterium of canker of kiwifruit in Japan[J]. Japanese Journal of Phytopathology, 1989, 55(4): 437-444.
[4]	李黎, 钟彩虹, 李大卫, 等. 猕猴桃细菌性溃疡病的研究进展[J]. 华中农业大学学报, 2013, 32(5): 124-133.
[5]	郭丽倩. 番茄斑驳花叶病毒和猕猴桃溃疡病菌单克隆抗体的创制及其检测应用[D]. 杭州: 浙江大学, 2020.
[6]	高小宁, 赵志博, 黄其玲, 等. 猕猴桃细菌性溃疡病研究进展[J]. 果树学报, 2012, 29(2): 262-268.
[7]	SCORTICHINI M, MARCELLETTI S, FERRANTE P, et al. Pseudomonas syringae pv. actinidiae: a re-emerging, multi-faceted, pandemic pathogen[J]. Molecular Plant Pathology, 2012, 13(7): 631-640.
[8]	朱海云, 安超, 李勃, 等. 猕猴桃溃疡病病原菌及检测方法研究进展[J]. 陕西农业科学, 2013, 59(4): 141-145, 153.
[9]	朱晓湘, 方炎祖, 廖新光. 猕猴桃溃疡病病原研究[J]. 湖南农业科学, 1993(6): 31-33.
[10]	SCORTICHINI M. Occurrence of Pseudomonas syringae pv. actinidiae on kiwifruit in Italy[J]. Plant Pathology, 1994, 43(6): 1035-1038.
[11]	KOH Y J, NOU I S. DNA markers for identification of Pseudomonas syringae pv. actinidiae[J]. Molecules and Cells, 2002, 13(2): 309-314.
[12]	ABELLEIRA A, LOPEZ M M, PENALVER J, et al. First report of bacterial canker of kiwifruit caused by Pseudomonas syringae pv. actinidiae in Spain[J]. Plant Disease, 2011, 95(12): 1583.
[13]	BASTAS K K, KARAKAYA A. First report of bacterial canker of kiwifruit caused by Pseudomonas syringae pv. actinidiae in Turkey[J]. Plant Disease, 2012, 96(3): 452.
[14]	DREO T, PIRC M, RAVNIKAR M, et al. First report of Pseudomonas syringae pv. actinidiae, the causal agent of bacterial canker of kiwifruit in Slovenia[J]. Plant Disease, 2014, 98(11): 1578.
[15]	VANNESTE J L, CORNISH D A, YU J, et al. First report of Pseudomonas syringae pv. actinidiae the causal agent of bacterial canker of kiwifruit on Actinidia arguta vines in new zealand[J]. Plant Disease, 2014, 98(3): 418.
[16]	HOLEVA M C, GLYNOS P E, KARAFLA C D. First report of bacterial canker of kiwifruit caused by Pseudomonas syringae pv. actinidiae in Greece[J]. Plant Disease, 2015, 99(5): 723.
[17]	JING Z B, YAO C C, LIU Z D. Isolation and identification of Pseudomonas syringae pv. actinidiae in Shaanxi Province, China[J]. Acta Horticulturae, 2018(1218): 279-286.
[18]	梁英梅, 张星耀, 田呈明, 等. 陕西省猕猴桃枝干溃疡病病原菌鉴定[J]. 西北林学院学报, 2000, 15(1): 37-39.
[19]	YANG X, YI X K, CHEN Y, et al. Identification of Pseudomonas syringae pv. actinidiae strains causing bacterial canker of kiwifruit in the Anhui Province of China, and determination of their streptomycin sensitivities[J]. Genetics and Molecular Research, 2015, 14(3): 8201-8210.
[20]	DAI Y, CHEN Y, GAN L, et al. First report of bacterial canker of kiwifruit caused by Pseudomonas syringae pv. actinidiae in Fujian Province, China[J]. Plant Disease, 2019, 103(1): 143.
[21]	鄢明峰. 奉新猕猴桃溃疡病病原菌鉴定、检测及室内药剂筛选[D]. 南昌: 江西农业大学, 2019.
[22]	CHAPMAN J R, TAYLOR R K, WEIR B S, et al. Phylogenetic relationships among global populations of Pseudomonas syringae pv. actinidiae[J]. Phytopathology, 2012, 102(11): 1034-1044.
[23]	朱海云, 李勃, 李燕, 等. 丁香假单胞菌猕猴桃致病变种的遗传多样性及进化关系[J]. 微生物学杂志, 2013, 33(4): 66-71.
[24]	VANNESTE J L, YU J, CORNISH D A, et al. Identification, virulence, and distribution of two biovars of Pseudomonas syringae pv. actinidiae in new Zealand[J]. Plant Disease, 2013, 97(6): 708-719.
[25]	CUNTY A, POLIAKOFF F, RIVOAL C, et al. Characterization of Pseudomonas syringae pv. actinidiae (P sa) isolated from France and assignment of Psa biovar 4 to a de novo pathovar: Pseudomonas syringae pv. actinidifoliorum pv. nov[J]. Plant Pathology, 2015, 64(3): 582-596.
[26]	SAWADA H, MIYOSHI T, IDE Y. Novel MLSA group (Psa5) of Pseudomonas syringae pv. actinidiae causing bacterial canker of kiwifruit (Actinidia chinensis) in Japan[J]. Japanese Journal of Phytopathology, 2014, 80(3): 171-184.
[27]	FUJIKAWA T, SAWADA H. Genome analysis of the kiwifruit canker pathogen Pseudomonas syringae pv. actinidiae biovar 5[J]. Scientific Reports, 2016, 6: 21399.
[28]	FARAHI L, GHAEMIMANESH F, MILANI S, et al. Monoclonal and polyclonal antibodies specific to human fibromodulin[J]. Iranian Journal of Biotechnology, 2019, 17(1): e2277.
[29]	邵宝林. 猕猴桃溃疡病风险分析及其病原鉴定检测和生物防治研究[D]. 雅安: 四川农业大学, 2013.
[30]	CIMMINO A, IANNACCONE M, PETRICCIONE M, et al. An ELISA method to identify the phytotoxic Pseudomonas syringae pv. actinidiae exopolysaccharides: a tool for rapid immunochemical detection of kiwifruit bacterial canker[J]. Phytochemistry Letters, 2017, 19: 136-140.
[31]	CHEN H, HU Y, QIN K Y, et al. A serological approach for the identification of the effector hopz5 of Pseudomonas syringae pv. actinidiae: a tool for the rapid immunodetection of kiwifruit bacterial canker[J]. Journal of Plant Pathology, 2018, 100(2): 171-177.
[32]	JUNG J S, HAN H S, JO Y S, et al. Nested PCR Detection of Pseudomonas syringae pv. actinidiae, the Causal Bacterium[J]. Research in Plant Disease, 2003, 9(3): 116-120.
[33]	VANNESTE J L. Recent progress on detecting understanding and controlling Pseudomonas syringae pv actinidiae a short review[J]. New Zealand Plant Protection, 2013, 66: 170-177.
[34]	REES-GEORGE J, VANNESTE J L, CORNISH D A, et al. Detection of Pseudomonas syringae pv. actinidiae using polymerase chain reaction (PCR) primers based on the 16S-23S rDNA intertranscribed spacer region and comparison with PCR primers based on other gene regions[J]. Plant Pathology, 2010, 59(3): 453-464.
[35]	SCORTICHINI M, MARCHESI U, DI PROSPERO P. Genetic relatedness among Pseudomonas avellanae, P. syringae pv. theae and P.s. pv. actinidiae, and their identification[J]. European Journal of Plant Pathology, 2002, 108(3): 269-278.
[36]	GALLELLI A, LAURORA A, LORETI S. Gene sequence analysis for the molecular detection of Pseudomonas syringae pv actinidiae developing diagnostic protocols[J]. Journal of Plant Pathology, 2011, 93(2): 425-435.
[37]	BIONDI E, GALEONE A, KUZMANOVI Ć N, et al. Pseudomonas syringae pv.actinidiae detection in kiwifruit plant tissue and bleeding sap[J]. Annals of Applied Biology, 2013, 162(1): 60-70.
[38]	邵宝林, 刘瑶, 朱天辉, 等. 猕猴桃溃疡病菌的分子检测技术研究[J]. 植物病理学报, 2013, 43(5): 458-466.
[39]	周大祥, 殷幼平, 王中康, 等. 利用EMA-qPCR建立快速检测猕猴桃溃疡病菌活菌的方法[J]. 植物保护, 2017, 43(3): 143-148.
[40]	刘芸宏. 猕猴桃Psa-V检测方法的研究[D]. 西安: 陕西师范大学, 2019.
[41]	GALLELLI A, TALOCCI S, PILOTTI M, et al. Real-time and qualitative PCR for detecting Pseudomonas syringae pv. actinidiae isolates causing recent outbreaks of kiwifruit bacterial canker[J]. Plant Pathology, 2014, 63(2): 264-276.
[42]	RUINELLI M, SCHNEEBERGER P H H, FERRANTE P, et al. Comparative genomics-informed design of two LAMP assays for detection of the kiwifruit pathogen Pseudomonas syringae pv. actinidiae and discrimination of isolates belonging to the pandemic biovar 3[J]. Plant Pathology, 2017, 66(1): 140-149.
[43]	王一波, 祝山, 郭文通, 等. 猕猴桃细菌性溃疡病可视化LAMP检测方法的建立与应用[J]. 西北农业学报, 2021, 30(5): 761-766.
[44]	雷庆, 叶华智, 余中树. 猕猴桃溃疡病菌噬菌体的初步研究[J]. 安徽农业科学, 2007, 35(19): 5995-5996.
[45]	FRAMPTON R A, TAYLOR C, HOLGUÍN MORENO A V, et al. Identification of bacteriophages for biocontrol of the kiwifruit canker phytopathogen Pseudomonas syringae pv. actinidiae[J]. Applied and Environmental Microbiology, 2014, 80(7): 2216-2228.
[46]	NI P E, WANG L, DENG B H, et al. Characterization of a lytic bacteriophage against Pseudomonas syringae pv. actinidiae and Its Endolysin[J]. Viruses, 2021, 13(4): 631.
[47]	PINHEIRO L A M, PEREIRA C, BARREAL M E, et al. Use of phage ϕ6 to inactivate Pseudomonas syringae pv. actinidiae in kiwifruit plants: in vitro and ex vivo experiments[J]. Applied Microbiology and Biotechnology, 2020, 104(3): 1319-1330.
[48]	SONG Y R, VU N T, PARK J, et al. Phage PPPL-1, A new biological agent to control bacterial canker caused by Pseudomonas syringae pv. actinidiae in kiwifruit[J]. Antibiotics, 2021, 10(5): 554.
[49]	YU J G, LIM J A, SONG Y R, et al. Isolation and characterization of bacteriophages against Pseudomonas syringae pv. actinidiae causing bacterial canker disease in kiwifruit[J]. Journal of Microbiology and Biotechnology, 2016, 26(2): 385-393.

检测方法	材料要求	适用场景	特异性	检测速度	工作量	检测成本
田间症状观察	活体植株	田间或实验室	低	慢	大	低
生物学测定	化学药剂	田间或实验室	中	慢	大	中
免疫学检测	酶标仪	田间或实验室	高	快	中	中
分子检测	PCR仪	实验室	高	快	中	中

检测方法	材料要求	适用场景	特异性	检测速度	工作量	检测成本
田间症状观察	活体植株	田间或实验室	低	慢	大	低
生物学测定	化学药剂	田间或实验室	中	慢	大	中
免疫学检测	酶标仪	田间或实验室	高	快	中	中
分子检测	PCR仪	实验室	高	快	中	中

检测技术	引物名称	序列方向	引物序列（5'-3'）	产物大小/bp	参考文献
KN-PCR	KNF	Forward	CACGATACATGGGCTTATGC	492	[11]
	KNR	Reverse	CTTTTCATCCACACACTCCG
RG-PCR	PsaF1	Forward	TTTTGCTTTGCACACCCGATTTT	280	[34]
	PsaR2	Reverse	CACGCACCCTTCAATCAGGATG	175
	PsaF3	Forward	ACCTGGTGAAGTTGGTCAGAGC
	PsaR4	Reverse	CGCACCCTTCAATCAGGATG
双重PCR	KNF	Forward	CACGATACATGGGCTTATGC	492	[36]
	KNR	Reverse	CTTTTCATCCACACACTCCG	226
	AvrDdpx-F	Forward	TTTCGGTGGTAACGTTGGCA
	AvrDdpx-R	Reverse	TTCCGCTAGGTGAAAAATGGG
巢氏PCR	cf1-1	Forward	GGCGCTCCCTCGCACTT	665	[32]
	cf1-2	Reverse	GGTATTGGCGGGGGTGC	310
	cf1-3	Forward	TCCTACGGTACGACGGAGTC
	cf1-4	Reverse	ACGGGGGATATGGAATAAGC
巢氏PCR	B1	Forward	GTGATTATCGGCACTGACGG	631	[37]
	B2	Reverse	CAAGCACACCATTGGTCATTGA	502
	KNF	Forward	CACGATACATGGGCTTATGC
	KNR	Reverse	CTTTTCATCCACACACTCCG
PCR	F7	Forward	CAATCATTTCGCCAGACGC	950	[38]
	R7	Reverse	CTACGAGGTTAGGTTCAGAGT
PCR-C	P0F	Forward	CTGCAACAGGCGACGGCGAGGC	243	[41]
	P6R	Reverse	CATAGGCTTCTGGTTTTCTTCCTGATCC
Real-time PCR-E	P3F	Forward	GGTTTCGGACACCGCAGGTTCTACCGAG	147
	P5R1	Reverse	CTTCCTGATCCCCGTTACCCATCGAC
Real-time PCR-F	P3F	Forward	GGTTTCGGACACCGCAGGTTCTACCGAG	158
	P6R	Reverse	CATAGGCTTCTGGTTTTCTTCCTGATCC
PSA LAMP	F3	Forward	GGCTCTCCTAGCAAGCATAC	无需电泳	[42]
	B3	Reverse	TGAGAAGGGACGCAACCA
	FIP	Forward	CGGATTCGCAACGCTCCAAGATCTGCTGAGCACGTTGGTC
	BIP	Reverse	ACTCTTCCGCAACGAGTTTGGGTCCCCACATGGAGTTGTCT
	loopF	Forward	TGCCGATCGAGTATCCATTTCCT
	loopR	Reverse	AGCCTTTTCCGAACGGTCT
PSA3 LAMP	F3	Forward	GCTATGGAATCCATTGCGGT
	B3	Reverse	CGCATCTGCTGGATCATCC
	FIP	Forward	ATCCCTTGCCCAGCACGAACATGAGGTCGAGGTGTCTGA
	BIP	Reverse	ATCCACAGTGGGTACACGGACGGGGGCACCTTCTTTCTTGG
	loopF	Forward	GGCCTTTTCAATGCGGTCAATATCC
	loopR	Reverse	GGTGTCAATCTGGTGGTGCAT
Psa5（Biovar 5）-PCR	Con002F	Forward	AACTCATACCCTGCGGTCAC	449	[27]
	Con002R	Reverse	GACACCGAGCAAAACCAAAT
	Con034F	Forward	CCAAACAACGTCTGGGCTAT	450
	Con034R	Reverse	TCGGCCTAGCTACGAGTGAT
	Con044F	Forward	AAGCGCCTTAATCTCGTTCA	470
	Con044R	Reverse	ATTCCGGATTGGGTATCACA
	Con067F	Forward	ATTTTAACGCCCATCTGCAC	439
	Con067R	Reverse	CTGCGGATTGCAACAGTCTA

检测技术	引物名称	序列方向	引物序列（5'-3'）	产物大小/bp	参考文献
KN-PCR	KNF	Forward	CACGATACATGGGCTTATGC	492	[11]
	KNR	Reverse	CTTTTCATCCACACACTCCG
RG-PCR	PsaF1	Forward	TTTTGCTTTGCACACCCGATTTT	280	[34]
	PsaR2	Reverse	CACGCACCCTTCAATCAGGATG	175
	PsaF3	Forward	ACCTGGTGAAGTTGGTCAGAGC
	PsaR4	Reverse	CGCACCCTTCAATCAGGATG
双重PCR	KNF	Forward	CACGATACATGGGCTTATGC	492	[36]
	KNR	Reverse	CTTTTCATCCACACACTCCG	226
	AvrDdpx-F	Forward	TTTCGGTGGTAACGTTGGCA
	AvrDdpx-R	Reverse	TTCCGCTAGGTGAAAAATGGG
巢氏PCR	cf1-1	Forward	GGCGCTCCCTCGCACTT	665	[32]
	cf1-2	Reverse	GGTATTGGCGGGGGTGC	310
	cf1-3	Forward	TCCTACGGTACGACGGAGTC
	cf1-4	Reverse	ACGGGGGATATGGAATAAGC
巢氏PCR	B1	Forward	GTGATTATCGGCACTGACGG	631	[37]
	B2	Reverse	CAAGCACACCATTGGTCATTGA	502
	KNF	Forward	CACGATACATGGGCTTATGC
	KNR	Reverse	CTTTTCATCCACACACTCCG
PCR	F7	Forward	CAATCATTTCGCCAGACGC	950	[38]
	R7	Reverse	CTACGAGGTTAGGTTCAGAGT
PCR-C	P0F	Forward	CTGCAACAGGCGACGGCGAGGC	243	[41]
	P6R	Reverse	CATAGGCTTCTGGTTTTCTTCCTGATCC
Real-time PCR-E	P3F	Forward	GGTTTCGGACACCGCAGGTTCTACCGAG	147
	P5R1	Reverse	CTTCCTGATCCCCGTTACCCATCGAC
Real-time PCR-F	P3F	Forward	GGTTTCGGACACCGCAGGTTCTACCGAG	158
	P6R	Reverse	CATAGGCTTCTGGTTTTCTTCCTGATCC
PSA LAMP	F3	Forward	GGCTCTCCTAGCAAGCATAC	无需电泳	[42]
	B3	Reverse	TGAGAAGGGACGCAACCA
	FIP	Forward	CGGATTCGCAACGCTCCAAGATCTGCTGAGCACGTTGGTC
	BIP	Reverse	ACTCTTCCGCAACGAGTTTGGGTCCCCACATGGAGTTGTCT
	loopF	Forward	TGCCGATCGAGTATCCATTTCCT
	loopR	Reverse	AGCCTTTTCCGAACGGTCT
PSA3 LAMP	F3	Forward	GCTATGGAATCCATTGCGGT
	B3	Reverse	CGCATCTGCTGGATCATCC
	FIP	Forward	ATCCCTTGCCCAGCACGAACATGAGGTCGAGGTGTCTGA
	BIP	Reverse	ATCCACAGTGGGTACACGGACGGGGGCACCTTCTTTCTTGG
	loopF	Forward	GGCCTTTTCAATGCGGTCAATATCC
	loopR	Reverse	GGTGTCAATCTGGTGGTGCAT
Psa5（Biovar 5）-PCR	Con002F	Forward	AACTCATACCCTGCGGTCAC	449	[27]
	Con002R	Reverse	GACACCGAGCAAAACCAAAT
	Con034F	Forward	CCAAACAACGTCTGGGCTAT	450
	Con034R	Reverse	TCGGCCTAGCTACGAGTGAT
	Con044F	Forward	AAGCGCCTTAATCTCGTTCA	470
	Con044R	Reverse	ATTCCGGATTGGGTATCACA
	Con067F	Forward	ATTTTAACGCCCATCTGCAC	439
	Con067R	Reverse	CTGCGGATTGCAACAGTCTA

猕猴桃溃疡病病菌的多样性及其检测技术

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