Bacterial artificial chromosome library construction of root-knot nematode resistant pepper genotype HDA149 and identification of clones linked to Me3 resistant locus
Pepper (Capsicum annuum. L.) is a widely cultivated vegetable crop worldwide and has the second largest planting area and the first largest vegetable output and value in China. Pepper root-knot nematode (Meloidogyne spp.) is one of the most serious pests of pepper, which caused huge losses every yea...
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| Format: | Article |
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KeAi Communications Co., Ltd.
2017-01-01
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| Series: | Journal of Integrative Agriculture |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2095311916614466 |
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| author | Xiao GUO Xiao-hui YANG Yu YANG Zhen-chuan MAO Feng LIU Wei-qing MA Bing-yan XIE Guang-cun LI |
| author_facet | Xiao GUO Xiao-hui YANG Yu YANG Zhen-chuan MAO Feng LIU Wei-qing MA Bing-yan XIE Guang-cun LI |
| author_sort | Xiao GUO |
| collection | DOAJ |
| description | Pepper (Capsicum annuum. L.) is a widely cultivated vegetable crop worldwide and has the second largest planting area and the first largest vegetable output and value in China. Pepper root-knot nematode (Meloidogyne spp.) is one of the most serious pests of pepper, which caused huge losses every year. Previous studies showed that the Me3 gene is resistant to a wide range of Meloidogyne species, including M. arenaria, M. javanica, and M. incognita. HDA149, a double haploid pepper genotype, harboring the root-knot nematode resistance gene Me3, was used to construct bacterial artificial chromosome library (BAC) via the vector of CopyControl™ pCC1 in this study. The library consists of 210 200 BAC clones and is equivalent to 5.3 pepper genomes. The average insert size is 95 kb, and most of them are 90–120 kb; but the empty clones are less than 3%. In order to screen the BAC library easily, 550 super pools with 384 BAC clones of each pool were further developed in this study. Specific primers from Me3 gene locus were used for BAC library screening, and more than 20 positive BAC clones were obtained. Then the selected positive BAC clones were analyzed by restriction enzyme digestion, BAC-end sequencing, marker development, and new positive BAC clones exploration, respectively. Finally, the contig with total length of about 300 kb linked to the Me3 locus was constructed based on chromosome walking strategy, which made a solid foundation for the cloning of the important root-knot nematode resistance gene Me3. |
| format | Article |
| id | doaj-art-e9eeaadebbd448a68549d97b2d3dfddf |
| institution | Matheson Library |
| issn | 2095-3119 |
| language | English |
| publishDate | 2017-01-01 |
| publisher | KeAi Communications Co., Ltd. |
| record_format | Article |
| series | Journal of Integrative Agriculture |
| spelling | doaj-art-e9eeaadebbd448a68549d97b2d3dfddf2025-08-02T22:26:18ZengKeAi Communications Co., Ltd.Journal of Integrative Agriculture2095-31192017-01-011615764Bacterial artificial chromosome library construction of root-knot nematode resistant pepper genotype HDA149 and identification of clones linked to Me3 resistant locusXiao GUO0Xiao-hui YANG1Yu YANG2Zhen-chuan MAO3Feng LIU4Wei-qing MA5Bing-yan XIE6Guang-cun LI7Institute of Vegetables and Flowers, Shandong Academy of Agricultural Sciences, Molecular Biology Key Laboratory of Shandong Facility Vegetable, Jinan 250100, P.R.China; GUO XiaoInstitute of Vegetables and Flowers, Shandong Academy of Agricultural Sciences, Molecular Biology Key Laboratory of Shandong Facility Vegetable, Jinan 250100, P.R.ChinaInstitute of Vegetables and Flowers, Shandong Academy of Agricultural Sciences, Molecular Biology Key Laboratory of Shandong Facility Vegetable, Jinan 250100, P.R.ChinaInstitute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.ChinaInstitute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.ChinaInstitute of Vegetables and Flowers, Shandong Academy of Agricultural Sciences, Molecular Biology Key Laboratory of Shandong Facility Vegetable, Jinan 250100, P.R.ChinaInstitute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China; Correspondence XIE Bing-yan, Tel: +86-10-82109546, Fax: +86-10-62174123Institute of Vegetables and Flowers, Shandong Academy of Agricultural Sciences, Molecular Biology Key Laboratory of Shandong Facility Vegetable, Jinan 250100, P.R.China; Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China; LI Guang-cun, Tel: +86-10-82105955, Fax: +86-10-62174123Pepper (Capsicum annuum. L.) is a widely cultivated vegetable crop worldwide and has the second largest planting area and the first largest vegetable output and value in China. Pepper root-knot nematode (Meloidogyne spp.) is one of the most serious pests of pepper, which caused huge losses every year. Previous studies showed that the Me3 gene is resistant to a wide range of Meloidogyne species, including M. arenaria, M. javanica, and M. incognita. HDA149, a double haploid pepper genotype, harboring the root-knot nematode resistance gene Me3, was used to construct bacterial artificial chromosome library (BAC) via the vector of CopyControl™ pCC1 in this study. The library consists of 210 200 BAC clones and is equivalent to 5.3 pepper genomes. The average insert size is 95 kb, and most of them are 90–120 kb; but the empty clones are less than 3%. In order to screen the BAC library easily, 550 super pools with 384 BAC clones of each pool were further developed in this study. Specific primers from Me3 gene locus were used for BAC library screening, and more than 20 positive BAC clones were obtained. Then the selected positive BAC clones were analyzed by restriction enzyme digestion, BAC-end sequencing, marker development, and new positive BAC clones exploration, respectively. Finally, the contig with total length of about 300 kb linked to the Me3 locus was constructed based on chromosome walking strategy, which made a solid foundation for the cloning of the important root-knot nematode resistance gene Me3.http://www.sciencedirect.com/science/article/pii/S2095311916614466pepperbacterial artificial chromosome library (BAC)root-knot nematodeMe3 genecontig |
| spellingShingle | Xiao GUO Xiao-hui YANG Yu YANG Zhen-chuan MAO Feng LIU Wei-qing MA Bing-yan XIE Guang-cun LI Bacterial artificial chromosome library construction of root-knot nematode resistant pepper genotype HDA149 and identification of clones linked to Me3 resistant locus Journal of Integrative Agriculture pepper bacterial artificial chromosome library (BAC) root-knot nematode Me3 gene contig |
| title | Bacterial artificial chromosome library construction of root-knot nematode resistant pepper genotype HDA149 and identification of clones linked to Me3 resistant locus |
| title_full | Bacterial artificial chromosome library construction of root-knot nematode resistant pepper genotype HDA149 and identification of clones linked to Me3 resistant locus |
| title_fullStr | Bacterial artificial chromosome library construction of root-knot nematode resistant pepper genotype HDA149 and identification of clones linked to Me3 resistant locus |
| title_full_unstemmed | Bacterial artificial chromosome library construction of root-knot nematode resistant pepper genotype HDA149 and identification of clones linked to Me3 resistant locus |
| title_short | Bacterial artificial chromosome library construction of root-knot nematode resistant pepper genotype HDA149 and identification of clones linked to Me3 resistant locus |
| title_sort | bacterial artificial chromosome library construction of root knot nematode resistant pepper genotype hda149 and identification of clones linked to me3 resistant locus |
| topic | pepper bacterial artificial chromosome library (BAC) root-knot nematode Me3 gene contig |
| url | http://www.sciencedirect.com/science/article/pii/S2095311916614466 |
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