Construction of Tomato Leaf Curl New Delhi Virus Infectious Clone and Its Application in Evaluation for Melon Disease Resistance

doi:10.16420/j.issn.0513-353x.2023-0733

Acta Horticulturae Sinica ›› 2024, Vol. 51 ›› Issue (11): 2555-2564.doi: 10.16420/j.issn.0513-353x.2023-0733

• Genetic & Breeding·Germplasm Resources·Molecular Biology • Previous Articles Next Articles

Construction of Tomato Leaf Curl New Delhi Virus Infectious Clone and Its Application in Evaluation for Melon Disease Resistance

LIU Liming¹, GENG Yanfei¹, HAO Fangmin², KANG Baoshan¹, WU Huijie¹, GU Qinsheng¹^,^*()

¹ Key Laboratory of Fruit Breeding Technology of Ministry of Agriculture and Rural Affairs，Zhengzhou Fruit Research Institute，Chinese Academy of Agricultural Sciences，Zhengzhou 450009，China
² Ningbo Key Laboratory of Characteristic Horticultural Crops in Quality Adjustment and Resistance Breeding，Ningbo，Zhejiang 315040，China

Received:2023-12-14 Revised:2024-08-20 Online:2024-12-12 Published:2024-11-25
Contact: GU Qinsheng

Abstract

Abstract:

In this study，a tomato leaf curl New Delhi virus（ToLCNDV）melon isolate Haimen 6 （HM6）was used as template to obtain its infectious clone by tandem repeat fragment method，and its infectivity was examined in different crops. Meanwhile，the resistance of 25 Cucumis melo cultivars were evaluated. The results showed that whatever infectious clones containing 1.57-mer DNA-A tandem repeat （1.57A）and 1.43-mer DNA-B tandem repeat（1.43B），respectively，were co-infiltrated or the clone containing both 1.57A and 1.43B was directly inoculated with melon，and the infection rate of the plants could reach 100%. Infectious clones containing 1.57A and 1.43B，respectively，were co-infiltrated into other crops，and it was found that it could infect Nicotiana benthamiana，Solanum lycopersicum，Cucurbita moschata，Cucurbita pepo，Lagenaria siceraria，Cucumis sativus，and Luffa aegyptiaca，but could not infect Citrullus lanatus. Through resistance evaluation，it was found that 3 of the 25 tested melon materials showed resistance，3 materials showed susceptibility and 19 materials showed high susceptibility.

Key words: melon, tomato leaf curl New Delhi virus, infectious clone, infectivity, resistance

LIU Liming, GENG Yanfei, HAO Fangmin, KANG Baoshan, WU Huijie, GU Qinsheng. Construction of Tomato Leaf Curl New Delhi Virus Infectious Clone and Its Application in Evaluation for Melon Disease Resistance[J]. Acta Horticulturae Sinica, 2024, 51(11): 2555-2564.

Figures/Tables 7

References 17

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用途 Usage	引物名称 Primer name	引物序列（5′-3′） Primer sequence
DNA-A扩增及侵染性克隆构建 Amplication and infectious clones construction of DNA-A	A-362F	ATTAGAGCTCGTGCAGTTGTCCCCATTG
	A-367R	TGACCATGATTACGAATTCGAGCTCCATAGGGGCTGTCGAAGTTG
	A-1555F	TAAAACGACGGCCAGTGCCAAGCTTGACCGGAATAGAGTGGTTCT
DNA-B扩增及侵染性克隆构建 Amplication and infectious clones construction of DNA-B	B-2020F	GCAGAAGCTTCTGCTCGAACATGG
	B-2060F	TGACCATGATTACGAATTCGAGCTCTGCAGCATCGTTTGTGAG
	B-2025R	TAAAACGACGGCCAGTGCCAAGCTTAGCCAGTTGAGAAATAG
	B-1520F	CGTTCGGACCTATAGCCAG
	B-518R	CTTGAGCTCTTGTTTCCGTTGTACGT
pToLCNDV-HM6构建 pToLCNDV-HM6 construction	1.43BF	AACCACTCTATTCCGGTCAAGCTTAATTCGAGCTCTTGTTTCCGTT
pToLCNDV-HM6构建 pToLCNDV-HM6 construction	1.43BR	ACGACGGCCAGTGCCAAGCTT
DNA-A检测Detection of DNA-A	ToLCNDV-A-61F	CCTATTTAATGCGTGGGGGT
DNA-A检测Detection of DNA-A	ToLCNDV-A-1622R	AGCACTGAAGCAGTGGGCAA
DNA-B检测Detection of DNA-B	ToLCNDV-B-62F	GCCGTTATCCGTCCGATTTG
DNA-B检测Detection of DNA-B	ToLCNDV-B-1598R	CGTCTTGTAGATGGTTCAAGG

用途 Usage	引物名称 Primer name	引物序列（5′-3′） Primer sequence
DNA-A扩增及侵染性克隆构建 Amplication and infectious clones construction of DNA-A	A-362F	ATTAGAGCTCGTGCAGTTGTCCCCATTG
	A-367R	TGACCATGATTACGAATTCGAGCTCCATAGGGGCTGTCGAAGTTG
	A-1555F	TAAAACGACGGCCAGTGCCAAGCTTGACCGGAATAGAGTGGTTCT
DNA-B扩增及侵染性克隆构建 Amplication and infectious clones construction of DNA-B	B-2020F	GCAGAAGCTTCTGCTCGAACATGG
	B-2060F	TGACCATGATTACGAATTCGAGCTCTGCAGCATCGTTTGTGAG
	B-2025R	TAAAACGACGGCCAGTGCCAAGCTTAGCCAGTTGAGAAATAG
	B-1520F	CGTTCGGACCTATAGCCAG
	B-518R	CTTGAGCTCTTGTTTCCGTTGTACGT
pToLCNDV-HM6构建 pToLCNDV-HM6 construction	1.43BF	AACCACTCTATTCCGGTCAAGCTTAATTCGAGCTCTTGTTTCCGTT
pToLCNDV-HM6构建 pToLCNDV-HM6 construction	1.43BR	ACGACGGCCAGTGCCAAGCTT
DNA-A检测Detection of DNA-A	ToLCNDV-A-61F	CCTATTTAATGCGTGGGGGT
DNA-A检测Detection of DNA-A	ToLCNDV-A-1622R	AGCACTGAAGCAGTGGGCAA
DNA-B检测Detection of DNA-B	ToLCNDV-B-62F	GCCGTTATCCGTCCGATTTG
DNA-B检测Detection of DNA-B	ToLCNDV-B-1598R	CGTCTTGTAGATGGTTCAAGG

编号 No.	甜瓜品种 Melon cultivar	病情指数 Disease index	抗性评价 Evaluation	编号 No.	甜瓜品种 Melon cultivar	病情指数 Disease index	抗性评价 Evaluation
1	绿秀 Lüxiu	100.0	高感 High sensitivity	14	众天翠宝 Zhongtian Cuibao	92.5	高感 High sensitivity
2	新密25号 Xinmi 25	100.0	高感 High sensitivity	15	博洋9号 Boyang 9	100.0	高感 High sensitivity
3	白玫 Baimei	100.0	高感 High sensitivity	16	甘甜3号 Gantian 3	77.5	高感 High sensitivity
4	黄梦脆 Huangmengcui	100.0	高感 High sensitivity	17	福甜一号 Futian 1	91.7	高感 High sensitivity
5	西州密25号 Xizhoumi 25	100.0	高感 High sensitivity	18	福甜二号 Futian 2	75.0	感病 Sensitivity
6	纳斯密 Nasimi	87.5	高感 High sensitivity	19	绿冠 Lüguan	100.0	高感 High sensitivity
7	雪蜜 Xuemi	100.0	高感 High sensitivity	20	早雪 Zaoxue	55.0	感病 Sensitivity
8	网络时代4号 Wangluo Shidai 4	100.0	高感 High sensitivity	21	锦烽 Jinfeng	75.0	感病 Sensitivity
9	玉姑 Yugu	100.0	高感 High sensitivity	22	Charentais	100.0	高感 High sensitivity
10	农友金香玉 Nongyou Jinxiangyu	80.6	高感 High sensitivity	23	PI414723	0	抗病 Resistance
11	新雅典 Xinyadian	100.0	高感 High sensitivity	24	PI124112	0	抗病 Resistance
12	众天翠玉 Zhongtian Cuiyu	100.0	高感 High sensitivity	25	PI179901	0	抗病 Resistance
13	皇妃 Huangfei	100.0	高感 High sensitivity	对照 Control	新密杂11号 Xinmiza 11	100.0	高感 High sensitivity

编号 No.	甜瓜品种 Melon cultivar	病情指数 Disease index	抗性评价 Evaluation	编号 No.	甜瓜品种 Melon cultivar	病情指数 Disease index	抗性评价 Evaluation
1	绿秀 Lüxiu	100.0	高感 High sensitivity	14	众天翠宝 Zhongtian Cuibao	92.5	高感 High sensitivity
2	新密25号 Xinmi 25	100.0	高感 High sensitivity	15	博洋9号 Boyang 9	100.0	高感 High sensitivity
3	白玫 Baimei	100.0	高感 High sensitivity	16	甘甜3号 Gantian 3	77.5	高感 High sensitivity
4	黄梦脆 Huangmengcui	100.0	高感 High sensitivity	17	福甜一号 Futian 1	91.7	高感 High sensitivity
5	西州密25号 Xizhoumi 25	100.0	高感 High sensitivity	18	福甜二号 Futian 2	75.0	感病 Sensitivity
6	纳斯密 Nasimi	87.5	高感 High sensitivity	19	绿冠 Lüguan	100.0	高感 High sensitivity
7	雪蜜 Xuemi	100.0	高感 High sensitivity	20	早雪 Zaoxue	55.0	感病 Sensitivity
8	网络时代4号 Wangluo Shidai 4	100.0	高感 High sensitivity	21	锦烽 Jinfeng	75.0	感病 Sensitivity
9	玉姑 Yugu	100.0	高感 High sensitivity	22	Charentais	100.0	高感 High sensitivity
10	农友金香玉 Nongyou Jinxiangyu	80.6	高感 High sensitivity	23	PI414723	0	抗病 Resistance
11	新雅典 Xinyadian	100.0	高感 High sensitivity	24	PI124112	0	抗病 Resistance
12	众天翠玉 Zhongtian Cuiyu	100.0	高感 High sensitivity	25	PI179901	0	抗病 Resistance
13	皇妃 Huangfei	100.0	高感 High sensitivity	对照 Control	新密杂11号 Xinmiza 11	100.0	高感 High sensitivity

Construction of Tomato Leaf Curl New Delhi Virus Infectious Clone and Its Application in Evaluation for Melon Disease Resistance

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