The fire blight resistance of apple clones in which DspE-interacting proteins are silenced

E. E. Borejsza-Wysocka, M. Malnoy, H. S. Aldwinckle, X. Meng, J. M. Bonasera, R. M. Nissinen, J. F. Kim, S. V. Beer

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Citations (Scopus)

Abstract

The disease specific gene, dspE (syn. dspA) of Erwinia amylovora, encodes a 198 kD pathogenicity effector that is essential for development of the fire blight disease in plants. Yeast two-hybrid and in vitro protein pull-down assays demonstrated that DspE interacts physically and specifically with four similar leucine-rich-repeat, receptor-like serine/theonine kinases from apple. Genes encoding the four DspE-interacting proteins of Malus (DIPM genes) are conserved in all hosts of E. amylovora tested, but not in tested non-host plants. The interaction between the DIPM's and DspE is thought to be involved in disease development. Sense sequences of ca. 400 bp from non-conserved regions of each DIPM gene, with homology among each other of <50%, were used to make constructs to be transformed into fire blight susceptible apple cultivars with the aim of silencing the DIPM genes and thereby preventing interaction with DspE. In addition, three constructs containing the four ca. 400 bp sequences of the individual genes in tandem, a full length sense sequence of one gene, and a "hairpin" sequence of that gene were made. All seven silencing sequences were transferred into 'Galaxy' apple cultivar by Agrobacterium inoculation of leaf pieces. Transgenic lines with all constructs were recovered. Assays of the transgenic plants for net expression of the target DIPM's by RT-PCR for their mRNA's have shown evidence of partial to complete silencing of some of the DIPM's in some transgenic lines. Evaluation for resistance to fire blight by inoculation of shoots with E. amylovora strain Ea273 indicated that some lines with silencing had increased resistance. Resistance due to silencing of a native apple gene(s) is likely to be more acceptable to regulators, growers and consumers than the use of heterologous.

Original languageEnglish
Title of host publicationProceedings of the Xth International Workshop on Fire Blight
EditorsC. Bazzi, U. Mazzucchi
Pages509-513
Number of pages5
Publication statusPublished - 2006 Feb 1

Publication series

NameActa Horticulturae
Volume704
ISSN (Print)0567-7572

Fingerprint

Erwinia amylovora
apples
clones
genes
proteins
genetically modified organisms
Malus
cultivars
assays
gene silencing
plant diseases and disorders
Agrobacterium
serine
leucine
transgenic plants
growers
phosphotransferases (kinases)
pathogenicity
reverse transcriptase polymerase chain reaction
yeasts

All Science Journal Classification (ASJC) codes

  • Horticulture

Cite this

Borejsza-Wysocka, E. E., Malnoy, M., Aldwinckle, H. S., Meng, X., Bonasera, J. M., Nissinen, R. M., ... Beer, S. V. (2006). The fire blight resistance of apple clones in which DspE-interacting proteins are silenced. In C. Bazzi, & U. Mazzucchi (Eds.), Proceedings of the Xth International Workshop on Fire Blight (pp. 509-513). (Acta Horticulturae; Vol. 704).
Borejsza-Wysocka, E. E. ; Malnoy, M. ; Aldwinckle, H. S. ; Meng, X. ; Bonasera, J. M. ; Nissinen, R. M. ; Kim, J. F. ; Beer, S. V. / The fire blight resistance of apple clones in which DspE-interacting proteins are silenced. Proceedings of the Xth International Workshop on Fire Blight. editor / C. Bazzi ; U. Mazzucchi. 2006. pp. 509-513 (Acta Horticulturae).
@inbook{0542aaa17cf64e85b5983cbaed8359f1,
title = "The fire blight resistance of apple clones in which DspE-interacting proteins are silenced",
abstract = "The disease specific gene, dspE (syn. dspA) of Erwinia amylovora, encodes a 198 kD pathogenicity effector that is essential for development of the fire blight disease in plants. Yeast two-hybrid and in vitro protein pull-down assays demonstrated that DspE interacts physically and specifically with four similar leucine-rich-repeat, receptor-like serine/theonine kinases from apple. Genes encoding the four DspE-interacting proteins of Malus (DIPM genes) are conserved in all hosts of E. amylovora tested, but not in tested non-host plants. The interaction between the DIPM's and DspE is thought to be involved in disease development. Sense sequences of ca. 400 bp from non-conserved regions of each DIPM gene, with homology among each other of <50{\%}, were used to make constructs to be transformed into fire blight susceptible apple cultivars with the aim of silencing the DIPM genes and thereby preventing interaction with DspE. In addition, three constructs containing the four ca. 400 bp sequences of the individual genes in tandem, a full length sense sequence of one gene, and a {"}hairpin{"} sequence of that gene were made. All seven silencing sequences were transferred into 'Galaxy' apple cultivar by Agrobacterium inoculation of leaf pieces. Transgenic lines with all constructs were recovered. Assays of the transgenic plants for net expression of the target DIPM's by RT-PCR for their mRNA's have shown evidence of partial to complete silencing of some of the DIPM's in some transgenic lines. Evaluation for resistance to fire blight by inoculation of shoots with E. amylovora strain Ea273 indicated that some lines with silencing had increased resistance. Resistance due to silencing of a native apple gene(s) is likely to be more acceptable to regulators, growers and consumers than the use of heterologous.",
author = "Borejsza-Wysocka, {E. E.} and M. Malnoy and Aldwinckle, {H. S.} and X. Meng and Bonasera, {J. M.} and Nissinen, {R. M.} and Kim, {J. F.} and Beer, {S. V.}",
year = "2006",
month = "2",
day = "1",
language = "English",
isbn = "9066054077",
series = "Acta Horticulturae",
pages = "509--513",
editor = "C. Bazzi and U. Mazzucchi",
booktitle = "Proceedings of the Xth International Workshop on Fire Blight",

}

Borejsza-Wysocka, EE, Malnoy, M, Aldwinckle, HS, Meng, X, Bonasera, JM, Nissinen, RM, Kim, JF & Beer, SV 2006, The fire blight resistance of apple clones in which DspE-interacting proteins are silenced. in C Bazzi & U Mazzucchi (eds), Proceedings of the Xth International Workshop on Fire Blight. Acta Horticulturae, vol. 704, pp. 509-513.

The fire blight resistance of apple clones in which DspE-interacting proteins are silenced. / Borejsza-Wysocka, E. E.; Malnoy, M.; Aldwinckle, H. S.; Meng, X.; Bonasera, J. M.; Nissinen, R. M.; Kim, J. F.; Beer, S. V.

Proceedings of the Xth International Workshop on Fire Blight. ed. / C. Bazzi; U. Mazzucchi. 2006. p. 509-513 (Acta Horticulturae; Vol. 704).

Research output: Chapter in Book/Report/Conference proceedingChapter

TY - CHAP

T1 - The fire blight resistance of apple clones in which DspE-interacting proteins are silenced

AU - Borejsza-Wysocka, E. E.

AU - Malnoy, M.

AU - Aldwinckle, H. S.

AU - Meng, X.

AU - Bonasera, J. M.

AU - Nissinen, R. M.

AU - Kim, J. F.

AU - Beer, S. V.

PY - 2006/2/1

Y1 - 2006/2/1

N2 - The disease specific gene, dspE (syn. dspA) of Erwinia amylovora, encodes a 198 kD pathogenicity effector that is essential for development of the fire blight disease in plants. Yeast two-hybrid and in vitro protein pull-down assays demonstrated that DspE interacts physically and specifically with four similar leucine-rich-repeat, receptor-like serine/theonine kinases from apple. Genes encoding the four DspE-interacting proteins of Malus (DIPM genes) are conserved in all hosts of E. amylovora tested, but not in tested non-host plants. The interaction between the DIPM's and DspE is thought to be involved in disease development. Sense sequences of ca. 400 bp from non-conserved regions of each DIPM gene, with homology among each other of <50%, were used to make constructs to be transformed into fire blight susceptible apple cultivars with the aim of silencing the DIPM genes and thereby preventing interaction with DspE. In addition, three constructs containing the four ca. 400 bp sequences of the individual genes in tandem, a full length sense sequence of one gene, and a "hairpin" sequence of that gene were made. All seven silencing sequences were transferred into 'Galaxy' apple cultivar by Agrobacterium inoculation of leaf pieces. Transgenic lines with all constructs were recovered. Assays of the transgenic plants for net expression of the target DIPM's by RT-PCR for their mRNA's have shown evidence of partial to complete silencing of some of the DIPM's in some transgenic lines. Evaluation for resistance to fire blight by inoculation of shoots with E. amylovora strain Ea273 indicated that some lines with silencing had increased resistance. Resistance due to silencing of a native apple gene(s) is likely to be more acceptable to regulators, growers and consumers than the use of heterologous.

AB - The disease specific gene, dspE (syn. dspA) of Erwinia amylovora, encodes a 198 kD pathogenicity effector that is essential for development of the fire blight disease in plants. Yeast two-hybrid and in vitro protein pull-down assays demonstrated that DspE interacts physically and specifically with four similar leucine-rich-repeat, receptor-like serine/theonine kinases from apple. Genes encoding the four DspE-interacting proteins of Malus (DIPM genes) are conserved in all hosts of E. amylovora tested, but not in tested non-host plants. The interaction between the DIPM's and DspE is thought to be involved in disease development. Sense sequences of ca. 400 bp from non-conserved regions of each DIPM gene, with homology among each other of <50%, were used to make constructs to be transformed into fire blight susceptible apple cultivars with the aim of silencing the DIPM genes and thereby preventing interaction with DspE. In addition, three constructs containing the four ca. 400 bp sequences of the individual genes in tandem, a full length sense sequence of one gene, and a "hairpin" sequence of that gene were made. All seven silencing sequences were transferred into 'Galaxy' apple cultivar by Agrobacterium inoculation of leaf pieces. Transgenic lines with all constructs were recovered. Assays of the transgenic plants for net expression of the target DIPM's by RT-PCR for their mRNA's have shown evidence of partial to complete silencing of some of the DIPM's in some transgenic lines. Evaluation for resistance to fire blight by inoculation of shoots with E. amylovora strain Ea273 indicated that some lines with silencing had increased resistance. Resistance due to silencing of a native apple gene(s) is likely to be more acceptable to regulators, growers and consumers than the use of heterologous.

UR - http://www.scopus.com/inward/record.url?scp=33745103427&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33745103427&partnerID=8YFLogxK

M3 - Chapter

AN - SCOPUS:33745103427

SN - 9066054077

SN - 9789066054073

T3 - Acta Horticulturae

SP - 509

EP - 513

BT - Proceedings of the Xth International Workshop on Fire Blight

A2 - Bazzi, C.

A2 - Mazzucchi, U.

ER -

Borejsza-Wysocka EE, Malnoy M, Aldwinckle HS, Meng X, Bonasera JM, Nissinen RM et al. The fire blight resistance of apple clones in which DspE-interacting proteins are silenced. In Bazzi C, Mazzucchi U, editors, Proceedings of the Xth International Workshop on Fire Blight. 2006. p. 509-513. (Acta Horticulturae).