Cellular Location of<i>Prune dwarf virus</i>in Almond Sections by In Situ Reverse Transcription-Polymerase Chain Reaction

Silva, Cristina; Tereso, Susana; Nolasco, Gustavo; Oliveira, M. Margarida

doi:10.1094/phyto.2003.93.3.278

Cited by 24 publications

(10 citation statements)

References 28 publications

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“…Samples from plantlets rooted using 0.005 or 0.4 mM IBA were collected from isolated roots along 10 d after hormonal induction. Tissues were fixed and sectioned following the procedure described by Silva et al (2003) for almond shoots. A general staining was performed by toluidine blue (Feder and O'Brien, 1968) for 5 min at 55 °C followed by washing with water and drying.…”

Section: Histology Of Adventitious Rootsmentioning

confidence: 99%

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Improved in vitro rooting of Prunus dulcis Mill. cultivars

Tereso¹,

Miguel²,

Mascarenhas³

et al. 2008

Biologia plant.

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A highly reproducible system was developed for efficient rooting of cultivars Boa Casta (BC) and Peneda and a BC seedling-derived clone (BC VII) of almond (Prunus dulcis Mill.). Twenty-four accessions derived from the clone BC VII and subjected to various in vitro culture treatments were screened. The long induction pre-treatment (LIP, 5 d), the brief induction pre-treatment (BIP, 16 h) and the hormonal shock by short dipping in hormone solution (1 min), were tested. BIP was the only that allowed rooting of cultivars. In BC VII, it induced high rooting frequencies (47 -100 %) when using a solution of 0.4 mM indole-3-butyric acid solidified with 2 g dm -3 gellam gum for 16-h. The response to the auxin type was variable depending on the cultivar and the root induction pre-treatment used. Root number was significantly different between the two cultivars and BC VII. Root length was significantly higher when using 0.005 mM IBA in LIP but this concentration induced apical necrosis. The improved acclimatization procedure for up to 4 weeks increased the survival to 45 %. The initiation and development of adventitious roots were proved to be asynchronous.Additional key words: adventitious root, indole-3-acetic acid, indole-3-butyric acid, mother plant age.

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Section: Histology Of Adventitious Rootsmentioning

confidence: 99%

“…The choice of the auxin type and concentration can differ according to the specific culture conditions involved. The rooting of some woody species including Prunus can be also improved under darkness during the first week (Sriskandarajah et al 1982, Rugini et al 1993, Caboni et al 1997.…”

Section: Introductionmentioning

confidence: 99%

Improved in vitro rooting of Prunus dulcis Mill. cultivars

Tereso¹,

Miguel²,

Mascarenhas³

et al. 2008

Biologia plant.

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“…Megagametophytes containing embryos and isolated zygotic embryos in diVerent developmental stages were Wxed and paraYn-embedded as described in Silva et al (2003), with the following modiWcation: tissues were Wxed in freshly prepared 4% paraformaldehyde and 0.25% glutaraldehyde pH 7.4 sodium phosphate buVer (PBS). In situ reverse-transcription PCR was generally performed as described in Silva et al (2003), with some modiWcations.…”

Section: Reverse Transcription Pcr (Rt-pcr)mentioning

confidence: 99%

“…In situ reverse-transcription PCR was generally performed as described in Silva et al (2003), with some modiWcations. After sectioning, samples were immersed in Histo-Clear for 10 min and then rehydrated through a graded ethanol series.…”

Section: Reverse Transcription Pcr (Rt-pcr)mentioning

confidence: 99%

PpRab1, a Rab GTPase from maritime pine is differentially expressed during embryogenesis

Gonçalves

Cairney

Rodríguez³

et al. 2007

Mol Genet Genomics

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Rab-related small GTP-binding proteins are known to be involved in the regulation of the vesicular transport system in eukaryotic cells. We report the characterization of a previously isolated full-length cDNA PpRab1 from Pinus pinaster. Amino acid sequence analysis revealed the presence of G1-G5 conserved domains of the GTPase Ras superfamily and a double cysteine motif in the C-terminal, characteristic of Rab proteins. The PpRab1 protein shows high sequence similarity to several Rab1 GTP-binding proteins in plants. Phylogenetic analysis showed that, within the Ras superfamily, PpRab1 is more closely related to the Rab family and within this, PpRab1 protein was found to cluster with Arabidopsis subfamily AtRABE, whose members are known to regulate ER-to-Golgi membrane trafficking steps. PpRab1 transcripts were expressed at constitutively high levels for the initial stages of zygotic embryo development, and then their relative abundance decreased as embryo matures. The PpRab1 transcript is not embryo-specific as it was found in roots, cotyledons and hypocotyls. An increase in PpRab1 expression level was observed when seeds are germinated and collected at successive time points of development. In situ RT-PCR analysis revealed an expression signal in early zygotic embryos. In view of the proposed roles of Rab1 GTP-binding protein, the possible function of the protein encoded by PpRab1 in embryogenesis is discussed.

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“…The symptoms of PDV infection vary through the year, and the severity of the infection is directly related to the orchard management. Ilarviruses are known to spread through pollen or seeds and recently it was shown that PDV is carried inside almond pollen grains (Silva et al 2003). Because most almond varieties are self-incompatible (Boskovic et al 1997), cross-pollination is required for fruit production, which complicates the control of the disease.…”

Section: Introductionmentioning

confidence: 99%

Expression of prune dwarf Ilarvirus coat protein sequences in Nicotiana benthamiana plants interferes with PDV systemic proliferation

Raquel

Lourenço

Moita³

et al. 2008

Plant Biotechnol Rep

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Prune dwarf virus (PDV) is an Ilarvirus systemically infecting almond trees and other Prunus species and spreading through pollen, among other means. We have studied strategies based on coat protein (cp) gene to block PDV replication in host plant cells. A Portuguese isolate of PDV was obtained from infected almond leaves and used to produce the cDNA of the cp gene. Various constructs were prepared based on this sequence, aiming for the transgenic expression of the original or modified PDV coat protein (cpPDVSense and cpPDVMutated) or for the expression of cpPDV RNA (cpPDVAntisense and cpPDVwithout start codon). All constructs were tested in a PDV host model, Nicotiana benthamiana, and extensive molecular characterization and controlled infections were performed on transformants and their progenies. Transgenic plants expressing the coat protein RNA were able to block the proliferation of a PDV isolate sharing only 91% homology with the isolate used for cpPDV cloning, as evaluated by DAS-ELISA on newly developed leaves. With cp expression, the blockage of PDV proliferation in newly developed leaves was only achieved with the construct cpPDV Mutated, where the coat protein has a substitution in the 14th aa residue, with arginine replaced by alanine. This result points to a possible role of the mutated amino acid in the virus ability to replicate and proliferate. This work reveals the possibility of achieving protection against PDV through either coat protein RNA or mutated cp sequence.

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Cellular Location ofPrune dwarf virusin Almond Sections by In Situ Reverse Transcription-Polymerase Chain Reaction

Cited by 24 publications

References 28 publications

Improved in vitro rooting of Prunus dulcis Mill. cultivars

Improved in vitro rooting of Prunus dulcis Mill. cultivars

PpRab1, a Rab GTPase from maritime pine is differentially expressed during embryogenesis

Expression of prune dwarf Ilarvirus coat protein sequences in Nicotiana benthamiana plants interferes with PDV systemic proliferation

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