新疆伽师县甜瓜病毒病种类鉴定

doi:10.16420/j.issn.0513-353x.2022-0543

摘要/Abstract

摘要：

于2019和2021年对新疆伽师县12个乡镇进行厚皮甜瓜（通常称伽师瓜）病毒病进行调研采样，采用两步法RT-PCR鉴定西瓜花叶病毒（watermelon mosaic virus，WMV）、黄瓜花叶病毒（cucumber mosaic virus，CMV）、小西葫芦黄化花叶病毒（zucchini yellow mosaic virus，ZYMV）、黄瓜绿斑驳花叶病毒（cucumber green mottle mosaic virus，CGMMV）、南瓜蚜传黄化病毒（cucurbit aphid-borne yellows virus，CABYV）和甜瓜蚜传黄化病毒（melon aphid-borne yellows virus，MABYV），并对WMV基因组进行分析。结果表明，伽师瓜病毒病发生较普遍，症状包括花叶、畸形、坏死和褪绿黄化等。采集的37份样品中，病毒的检出率为21.6%（8/37），其中WMV的检出率最高，为18.9%（7/37），CMV和ZYMV次之，没有鉴定到CGMMV、CABYV和MABYV，复合侵染更普遍。不同WMV分离物的F2/R2（P1 + HC-Pro）、F4/R4（P3 + 6K1 + CI）、F5/R5（CI + 6K2 + NIa-VPg）、F6/R6（6K2 + NIa-VPg + NIa-Pro + NIb）和F8/R8（CP + 3′-UTR）等基因组片段在进化树上的分支具有一定的差异，表明WMV基因组可能存在重组现象。

关键词: 甜瓜, 病毒鉴定, 基因组分析, 西瓜花叶病毒

Abstract:

In 2019 and 2021,12 towns in Jiashi county of Xinjiang were investigated and sampled for muskmelon（Jiashi-melon）virus disease. Watermelon mosaic virus（WMV），cucumber mosaic virus（CMV），zucchini yellow mosaic virus（ZYMV），cucumber green mottle mosaic virus（CGMMV），cucurbit aphid-borne yellows virus（CABYV），and melon aphid-borne yellows virus（MABYV）were identified by two-step RT-PCR protocol. In addition，the genome of WMV isolates was analyzed. The results showed that the virus disease is more common in the Jiashi-melon planting towns in Jiashi County. Symptoms include mosaic，blister mosaic，blister mosaic and malformation，leaf necrosis，leaf chlorosis，and fruit malformation. A total of 37 samples were collected，and the detection of WMV，CMV，and ZYMV was positive，but CGMMV，CABYV，and MABYV were not detected. 21.6%（8/37）of the samples were positive. WMV was the most prevalent virus detected in 18.9%（7/37）of the 37 samples collected，followed by CMV and ZYMV. Multiple infections were more frequent than single infections. F2/R2（P1 + HC-Pro），F4/R4（P3 + 6K1 + CI），F5/R5（CI + 6K2 + NIa-VPg），F6/R6（6K2 + NIa-VPg + NIa-Pro + NIb），and F8/R8（CP + 3′-UTR）fragments of different WMV isolates infecting Jiashi-melon have certain differences in the branches of the evolutionary tree，indicating that there may be recombination in the WMV genome.

Key words: melon, virus identification, genome analysis, watermelon mosaic virus

王东, 麦麦提艾则孜·穆合塔尔, 刘艳全, 迪力达尔克孜·吾拉木. 新疆伽师县甜瓜病毒病种类鉴定[J]. 园艺学报, 2023, 50(8): 1793-1802.

WANG Dong, MAIMAITIAIZEZI · Muhetaer, LIU Yanquan, DILIDAERKEZI · Wulamu. Identification of Virus Disease Types of Melon Planting in Jiashi County of Xinjiang[J]. Acta Horticulturae Sinica, 2023, 50(8): 1793-1802.

图/表 6

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病毒 Virus	正向引物（5′-3′） Forward primer	反向引物（5′-3′） Reverse primer	参考文献 Reference
WMV	WMV-F：GGTTGCTGYGARTCAGTGTC	WMV-R：CGACCCGAAATGCTAACT	刘卫荣和向本春，2008
CMV	CMVAu1u：CATGGATGCTTCTCCRCGAG	CMVAu2d：CGTAAGCTGGATGGACAACC	Stanković et al.，2011
ZYMV	CPfwd：GCTCCATACATAGCTGAGAC	Cprev：AACGGAGTCTAATCTCGAGC	Pfosser & Baumann,2002
CGMMV	6F：GATGCATTCTGTTGCAGAGG	6R：TCAACCTCACACACGTAAGAGG	Ali et al.,2004 Shang et al.，2012;吴洋等，2017a
CABYV	Pocon F：TGYTCYGGTTTTGACTGG CA3414F：GGATTTTCTAGCGGTCTA	Pococp R：CGTCTACCTATTTSGGRTTN Pococp R：CGTCTACCTATTTSGGRTTN	Ali et al.,2004 Shang et al.，2012;吴洋等，2017a
MABYV	Pocon F：TGYTCYGGTTTTGACTGG	Pococp R：CGTCTACCTATTTSGGRTTN	Shang et al.，2012;吴洋等，2017a
	MA3669F：CGACGACGACGAAATCCAA	Pococp R;CGTCTACCTATTTSGGRTTN	Shang et al.，2012;吴洋等，2017a

病毒 Virus	正向引物（5′-3′） Forward primer	反向引物（5′-3′） Reverse primer	参考文献 Reference
WMV	WMV-F：GGTTGCTGYGARTCAGTGTC	WMV-R：CGACCCGAAATGCTAACT	刘卫荣和向本春，2008
CMV	CMVAu1u：CATGGATGCTTCTCCRCGAG	CMVAu2d：CGTAAGCTGGATGGACAACC	Stanković et al.，2011
ZYMV	CPfwd：GCTCCATACATAGCTGAGAC	Cprev：AACGGAGTCTAATCTCGAGC	Pfosser & Baumann,2002
CGMMV	6F：GATGCATTCTGTTGCAGAGG	6R：TCAACCTCACACACGTAAGAGG	Ali et al.,2004 Shang et al.，2012;吴洋等，2017a
CABYV	Pocon F：TGYTCYGGTTTTGACTGG CA3414F：GGATTTTCTAGCGGTCTA	Pococp R：CGTCTACCTATTTSGGRTTN Pococp R：CGTCTACCTATTTSGGRTTN	Ali et al.,2004 Shang et al.，2012;吴洋等，2017a
MABYV	Pocon F：TGYTCYGGTTTTGACTGG	Pococp R：CGTCTACCTATTTSGGRTTN	Shang et al.，2012;吴洋等，2017a
	MA3669F：CGACGACGACGAAATCCAA	Pococp R;CGTCTACCTATTTSGGRTTN	Shang et al.，2012;吴洋等，2017a

引物名称 Name	引物序列（5′-3′） Sequence	扩增片段位置 Position	覆盖的基因组区域 Region
F1/R1	ACACGAATGGCTACAAT/TGTACTCCTCCCAACTAA	127 ~ 1 678	P1 + HC-Pro
F2/R2	TTTGTGGTGCGAGGTAGA/AAGCCTTCCCATCGTCAT	1 383 ~ 2 368	P1 + HC-Pro
F3/R3	CTGAGGGATACAGTAAATACG/AGCCATGAATGCAACAA	2 150 ~ 3 927	HC-Pro + P3 + 6K1
F4/R4	TCCCGTAGCAAAGAGC/GCATATCGGCGTTTGA	3 828 ~ 4 765 4 746 ~ 6 140	P3 + 6K1 + CI
F5/R5	AGGCTCAAACGCCGATAT/CTTCACGCCCTACTTTCC	3 828 ~ 4 765 4 746 ~ 6 140	CI + 6K2 + NIa-VPg
F6/R6	GGATGCTGTGGGACTACTT/TCAATGGCTCAAAGAACT	6 041 ~ 7 558 7 433 ~ 9 012	6K2 + NIa-VPg + NIa-Pro + NIb
F7/R7	ATTTGGTGGCTTGTGGGC/GCCACTGGTGTCCTTCTTCG	6 041 ~ 7 558 7 433 ~ 9 012	NIb + CP
F8/R8	ATTTGGATGCAGGAAAGG/AGGACAACAAACATTACCGT	8 972 ~ 10 046	CP + 3′-UTR

引物名称 Name	引物序列（5′-3′） Sequence	扩增片段位置 Position	覆盖的基因组区域 Region
F1/R1	ACACGAATGGCTACAAT/TGTACTCCTCCCAACTAA	127 ~ 1 678	P1 + HC-Pro
F2/R2	TTTGTGGTGCGAGGTAGA/AAGCCTTCCCATCGTCAT	1 383 ~ 2 368	P1 + HC-Pro
F3/R3	CTGAGGGATACAGTAAATACG/AGCCATGAATGCAACAA	2 150 ~ 3 927	HC-Pro + P3 + 6K1
F4/R4	TCCCGTAGCAAAGAGC/GCATATCGGCGTTTGA	3 828 ~ 4 765 4 746 ~ 6 140	P3 + 6K1 + CI
F5/R5	AGGCTCAAACGCCGATAT/CTTCACGCCCTACTTTCC	3 828 ~ 4 765 4 746 ~ 6 140	CI + 6K2 + NIa-VPg
F6/R6	GGATGCTGTGGGACTACTT/TCAATGGCTCAAAGAACT	6 041 ~ 7 558 7 433 ~ 9 012	6K2 + NIa-VPg + NIa-Pro + NIb
F7/R7	ATTTGGTGGCTTGTGGGC/GCCACTGGTGTCCTTCTTCG	6 041 ~ 7 558 7 433 ~ 9 012	NIb + CP
F8/R8	ATTTGGATGCAGGAAAGG/AGGACAACAAACATTACCGT	8 972 ~ 10 046	CP + 3′-UTR

采用地点 Location	症状表现 Symptom
英买里乡Yingmaili Township	花叶、叶片坏死、叶片褪绿黄化 Mosaic，leaf necrosis，leaf chlorosis
江巴孜乡Gambazi Township	花叶、叶片坏死、叶片褪绿黄化 Mosaic，leaf necrosis，leaf chlorosis
卧里托格拉克镇Wolitogorak Township	花叶、疱斑花叶、叶片坏死、叶片褪绿黄化 Mosaic，blister mosaic，leaf necrosis，leaf chlorosis
克孜勒博依乡Kizilboyi Township	花叶、疱斑花叶、疱斑花叶畸形、叶片坏死、叶片褪绿黄化、果实畸形 Mosaic，blister mosaic，blister mosaic and malformation，leaf necrosis，leaf chlorosis，fruit malformation
米夏乡Mixia Township	花叶、叶片坏死、叶片褪绿黄化 Mosaic，leaf necrosis，leaf chlorosis
夏普吐勒乡Shartuole Township	叶片坏死、叶片褪绿黄化 Leaf necrosis，leaf chlorosis
和夏阿瓦提乡Hexiaavati Township	花叶、叶片褪绿黄化 Mosaic，leaf chlorosis
克孜勒苏乡Kizilsu Township	花叶、疱斑花叶、叶片褪绿黄化 Mosaic，blister mosaic，leaf chlorosis
古勒鲁克乡Guleruk Township	花叶、疱斑花叶、叶片坏死、叶片褪绿黄化 Mosaic，blister mosaic，leaf necrosis，leaf chlorosis
玉代克力克乡Yudaikelik Township	花叶、叶片坏死、叶片褪绿黄化 Mosaic，leaf necrosis，leaf chlorosis
巴仁镇Baren Township	花叶、叶片坏死 Mosaic，leaf necrosis
铁日木乡Tierimu Township	疱斑花叶、叶片褪绿黄化 Blister mosaic，leaf chlorosis