Abstract
| Original language | English |
|---|---|
| Pages (from-to) | 579-591 |
| Journal | Plant Molecular Biology |
| Volume | 75 |
| Issue number | 6 |
| DOIs | |
| Publication status | Published - 2011 |
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Keywords
- receptor-like genes
- real-time pcr
- venturia-inaequalis
- vf gene
- plants
- sequences
- agrobacterium
- promoters
- linkage
- cluster
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Functional analysis and expression of HcrVf1 and HcrVf2 for development of scab resistant cisgenic and intragenic apples. / Joshi, S.G.; Schaart, J.; Groenwold, R.; Jacobsen, E.; Schouten, H.J.; Krens, F.A.
In: Plant Molecular Biology, Vol. 75, No. 6, 2011, p. 579-591.Research output: Contribution to journal › Article › Academic › peer-review
TY - JOUR
T1 - Functional analysis and expression of HcrVf1 and HcrVf2 for development of scab resistant cisgenic and intragenic apples
AU - Joshi, S.G.
AU - Schaart, J.
AU - Groenwold, R.
AU - Jacobsen, E.
AU - Schouten, H.J.
AU - Krens, F.A.
PY - 2011
Y1 - 2011
N2 - Apple scab resistance genes, HcrVf1 and HcrVf2, were isolated including their native promoter, coding and terminator sequences. Two fragment lengths (short and long) of the native gene promoters and the strong apple rubisco gene promoter (PMdRbc) were used for both HcrVf genes to test their effect on expression and phenotype. The scab susceptible cultivar ‘Gala’ was used for plant transformations and after selection of transformants, they were micrografted onto apple seedling rootstocks for scab disease tests. Apple transformants were also tested for HcrVf expression by quantitative RT-PCR (qRTPCR). For HcrVf1 the long native promoter gave significantly higher expression that the short one; in case of HcrVf2 the difference between the two was not significant. The apple rubisco gene promoter proved to give the highest expression of both HcrVf1 and HcrVf2. The top four expanding leaves were used initially for inoculation with monoconidial isolate EU-B05 which belongs to race 1 of V. inaequalis. Later six other V. inaequalis isolates were used to study the resistance spectra of the individual HcrVf genes. The scab disease assays showed that HcrVf1 did not give resistance against any of the isolates tested regardless of the expression level. The HcrVf2 gene appeared to be the only functional gene for resistance against Vf avirulent isolates of V. inaequalis. HcrVf2 did not provide any resistance to Vf virulent strains, even not in case of overexpression. In conclusion, transformants carrying the apple-derived HcrVf2 gene in a cisgenic as well as in an intragenic configuration were able to reach scab resistance levels comparable to the Vf resistant control cultivar obtained by classical breeding, cv. ‘Santana’.
AB - Apple scab resistance genes, HcrVf1 and HcrVf2, were isolated including their native promoter, coding and terminator sequences. Two fragment lengths (short and long) of the native gene promoters and the strong apple rubisco gene promoter (PMdRbc) were used for both HcrVf genes to test their effect on expression and phenotype. The scab susceptible cultivar ‘Gala’ was used for plant transformations and after selection of transformants, they were micrografted onto apple seedling rootstocks for scab disease tests. Apple transformants were also tested for HcrVf expression by quantitative RT-PCR (qRTPCR). For HcrVf1 the long native promoter gave significantly higher expression that the short one; in case of HcrVf2 the difference between the two was not significant. The apple rubisco gene promoter proved to give the highest expression of both HcrVf1 and HcrVf2. The top four expanding leaves were used initially for inoculation with monoconidial isolate EU-B05 which belongs to race 1 of V. inaequalis. Later six other V. inaequalis isolates were used to study the resistance spectra of the individual HcrVf genes. The scab disease assays showed that HcrVf1 did not give resistance against any of the isolates tested regardless of the expression level. The HcrVf2 gene appeared to be the only functional gene for resistance against Vf avirulent isolates of V. inaequalis. HcrVf2 did not provide any resistance to Vf virulent strains, even not in case of overexpression. In conclusion, transformants carrying the apple-derived HcrVf2 gene in a cisgenic as well as in an intragenic configuration were able to reach scab resistance levels comparable to the Vf resistant control cultivar obtained by classical breeding, cv. ‘Santana’.
KW - receptor-like genes
KW - real-time pcr
KW - venturia-inaequalis
KW - vf gene
KW - plants
KW - sequences
KW - agrobacterium
KW - promoters
KW - linkage
KW - cluster
U2 - 10.1007/s11103-011-9749-1
DO - 10.1007/s11103-011-9749-1
M3 - Article
VL - 75
SP - 579
EP - 591
JO - Plant Molecular Biology
JF - Plant Molecular Biology
SN - 0167-4412
IS - 6
ER -