Vector integration in triple R gene transformants and the clustered inheritance of resistance against potato late blight

S. Zhu, A. Duwal, Q. Su, J.H. Vossen, R.G.F. Visser, E. Jacobsen

Research output: Contribution to journalArticleAcademicpeer-review

11 Citations (Scopus)

Abstract

Genetic transformation with resistance (R) genes is expected to enhance resistance durability against pathogens, especially for potato, a vegetatively propagated crop with tetrasomic inheritance and a long-term breeding program. In this study, 128 potato transformants were analysed for the presence of vector T-DNA genes, borders and backbone sequences. They were harvested after transformation using a construct containing neomycin phosphotransferase II (nptII) and three R genes against potato late blight (Phytophthora infestans). Our analysis revealed that 45 % of the R gene-containing transformants possessed a low T-DNA copy number, without the integration of vector backbone and borders. The integration of vector backbone sequences was characterized using eight genes, and backbone gene tetA was selected for the early prediction of plants with backbone sequence integration. Three transformants, two plants harbouring one T-DNA copy and one plant harbouring three T-DNA copies, were crossed with susceptible cv. Katahdin. Based on our results, we conclude that all four T-DNA genes were inherited as one cluster and segregated in a Mendelian fashion. The three T-DNA inserts from the transformant harbouring three T-DNA copies were statistically proven to be un-linked and inherited into the offspring plants independently. All of the R genes were functionally expressed in the offspring plants as in their parental transformants. This functional gene stacking has important implications towards achieving more durable resistance against potato late blight
Original languageEnglish
Pages (from-to)315-325
JournalTransgenic Research
Volume22
Issue number2
DOIs
Publication statusPublished - 2013

Fingerprint

vpr Genes
Phytophthora infestans
Solanum tuberosum
inheritance (genetics)
DNA
genes
Genes
Kanamycin Kinase
Genetic Transformation
potatoes
tetrasomics
T-DNA
kanamycin kinase
Breeding
genetic transformation
durability

Keywords

  • agrobacterium-mediated transformation
  • phytophthora-infestans
  • backbone sequences
  • transgene expression
  • plant transformation
  • dna transfer
  • genome
  • rice
  • tumefaciens
  • chromosome

Cite this

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title = "Vector integration in triple R gene transformants and the clustered inheritance of resistance against potato late blight",
abstract = "Genetic transformation with resistance (R) genes is expected to enhance resistance durability against pathogens, especially for potato, a vegetatively propagated crop with tetrasomic inheritance and a long-term breeding program. In this study, 128 potato transformants were analysed for the presence of vector T-DNA genes, borders and backbone sequences. They were harvested after transformation using a construct containing neomycin phosphotransferase II (nptII) and three R genes against potato late blight (Phytophthora infestans). Our analysis revealed that 45 {\%} of the R gene-containing transformants possessed a low T-DNA copy number, without the integration of vector backbone and borders. The integration of vector backbone sequences was characterized using eight genes, and backbone gene tetA was selected for the early prediction of plants with backbone sequence integration. Three transformants, two plants harbouring one T-DNA copy and one plant harbouring three T-DNA copies, were crossed with susceptible cv. Katahdin. Based on our results, we conclude that all four T-DNA genes were inherited as one cluster and segregated in a Mendelian fashion. The three T-DNA inserts from the transformant harbouring three T-DNA copies were statistically proven to be un-linked and inherited into the offspring plants independently. All of the R genes were functionally expressed in the offspring plants as in their parental transformants. This functional gene stacking has important implications towards achieving more durable resistance against potato late blight",
keywords = "agrobacterium-mediated transformation, phytophthora-infestans, backbone sequences, transgene expression, plant transformation, dna transfer, genome, rice, tumefaciens, chromosome",
author = "S. Zhu and A. Duwal and Q. Su and J.H. Vossen and R.G.F. Visser and E. Jacobsen",
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Vector integration in triple R gene transformants and the clustered inheritance of resistance against potato late blight. / Zhu, S.; Duwal, A.; Su, Q.; Vossen, J.H.; Visser, R.G.F.; Jacobsen, E.

In: Transgenic Research, Vol. 22, No. 2, 2013, p. 315-325.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Vector integration in triple R gene transformants and the clustered inheritance of resistance against potato late blight

AU - Zhu, S.

AU - Duwal, A.

AU - Su, Q.

AU - Vossen, J.H.

AU - Visser, R.G.F.

AU - Jacobsen, E.

PY - 2013

Y1 - 2013

N2 - Genetic transformation with resistance (R) genes is expected to enhance resistance durability against pathogens, especially for potato, a vegetatively propagated crop with tetrasomic inheritance and a long-term breeding program. In this study, 128 potato transformants were analysed for the presence of vector T-DNA genes, borders and backbone sequences. They were harvested after transformation using a construct containing neomycin phosphotransferase II (nptII) and three R genes against potato late blight (Phytophthora infestans). Our analysis revealed that 45 % of the R gene-containing transformants possessed a low T-DNA copy number, without the integration of vector backbone and borders. The integration of vector backbone sequences was characterized using eight genes, and backbone gene tetA was selected for the early prediction of plants with backbone sequence integration. Three transformants, two plants harbouring one T-DNA copy and one plant harbouring three T-DNA copies, were crossed with susceptible cv. Katahdin. Based on our results, we conclude that all four T-DNA genes were inherited as one cluster and segregated in a Mendelian fashion. The three T-DNA inserts from the transformant harbouring three T-DNA copies were statistically proven to be un-linked and inherited into the offspring plants independently. All of the R genes were functionally expressed in the offspring plants as in their parental transformants. This functional gene stacking has important implications towards achieving more durable resistance against potato late blight

AB - Genetic transformation with resistance (R) genes is expected to enhance resistance durability against pathogens, especially for potato, a vegetatively propagated crop with tetrasomic inheritance and a long-term breeding program. In this study, 128 potato transformants were analysed for the presence of vector T-DNA genes, borders and backbone sequences. They were harvested after transformation using a construct containing neomycin phosphotransferase II (nptII) and three R genes against potato late blight (Phytophthora infestans). Our analysis revealed that 45 % of the R gene-containing transformants possessed a low T-DNA copy number, without the integration of vector backbone and borders. The integration of vector backbone sequences was characterized using eight genes, and backbone gene tetA was selected for the early prediction of plants with backbone sequence integration. Three transformants, two plants harbouring one T-DNA copy and one plant harbouring three T-DNA copies, were crossed with susceptible cv. Katahdin. Based on our results, we conclude that all four T-DNA genes were inherited as one cluster and segregated in a Mendelian fashion. The three T-DNA inserts from the transformant harbouring three T-DNA copies were statistically proven to be un-linked and inherited into the offspring plants independently. All of the R genes were functionally expressed in the offspring plants as in their parental transformants. This functional gene stacking has important implications towards achieving more durable resistance against potato late blight

KW - agrobacterium-mediated transformation

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KW - dna transfer

KW - genome

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KW - tumefaciens

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SN - 0962-8819

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ER -