Rooting and Acclimatization of In Vitro-grown Shoots from Mature Trees of Three Persian Walnut Cultivars

Vahdati, Kourosh; Leslie, Charles A.; Zamani, Zabihollah; McGranahan, Gale H.

doi:10.21273/hortsci.39.2.324

Cited by 47 publications

(35 citation statements)

References 13 publications

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“…In vitro shoot cuttings were maintained on MS basal medium (Murashige and Skoog 1962) supplemented with 2% (w/ v) sucrose (MS2) and used as the source of leaf explants. Organized embryogenic structures (OES) were induced from immature leaves 2-5 mm in length by culturing for 4 wk on DKW/Juglans basal medium (Vahdati et al 2004) supplemented with 2% (w/v) sucrose, MS vitamins, and 50 μM picloram. OES was excised and used to establish FEC target tissues through three successive culture cycles on Gresshoff and Doy basal medium (Gresshoff and Doy 1972) supplemented with 2% (w/v) sucrose and 50 μM picloram (GD2 50P) (Taylor et al 2001(Taylor et al , 2012.…”

Section: Methodsmentioning

confidence: 99%

Evaluation of red fluorescent protein (DsRed) as alternative visual marker of genetic transformation in cassava (Manihot esculenta Crantz)

Okwuonu

Achi

Egesi

et al. 2015

In Vitro Cell.Dev.Biol.-Plant

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Red fluorescent protein (DsRed) from reef coral was evaluated in comparison with green fluorescent protein (GFP) as a reporter gene for cassava transformation. Cassava friable embryogenic callus (FEC) was transformed with ERtargeted versions of DsRed and GFP constructs driven by the 35S cauliflower mosaic virus promoter. Efficiency of transformation was comparable for both visual marker genes at averages of 119 and 163 expressing plants recovered per cc of settled cell volume FEC for GFP and DsRed, respectively. High and uniform DsRed expression was observed at the single cell and proliferating callus stages, in somatic embryos and within organs of whole in vitro and greenhouse-grown plants in a manner similar to GFP. Plants expressing GFP and DsRed were robust and phenotypically normal with regard to growth, vigor, and formation of storage roots when grown in the greenhouse. Expression of marker genes within cross sections of petiole, woody stem, and storage roots from greenhousegrown plants was determined. The interference of phenolic compounds and chlorotic tissues characteristic of the signal from GFP-expressing tissues was not observed within tissues transgenic for DsRed. Tissues and plants co-expressing DsRed and GFP were produced by co-culturing FEC with a mixed Agrobacterium suspension carrying GFP and DsRed gene constructs or by re-transformation of an existing GFP transgenic line with DsRed. Re-transformation of GFPexpressing tissues was the more efficient method for production of GFP/DsRed stacked plants. Co-expression of both marker genes within the same transformation unit was easily visualized at their respective wavelength with the aid of appropriate filters thus validating their potential for coexpression studies.

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Section: Methodsmentioning

confidence: 99%

Evaluation of red fluorescent protein (DsRed) as alternative visual marker of genetic transformation in cassava (Manihot esculenta Crantz)

Okwuonu

Achi

Egesi

et al. 2015

In Vitro Cell.Dev.Biol.-Plant

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“…The latter is used in virtually all in vitro walnut protocols (e.g., McGranahan and Leslie 1988;Jay-Allemand et al 1992;Dolcet-Sanjuan et al 2004;Vahdati et al 2004;Leslie et al 2009). No clear pattern of influence on rhizogenesis emerged from these experiments, where genotype was identified as the major driver of in vitro behavior (it explains 76.5 % of the total variance).…”

Section: Effects Of the Carbon Sourcementioning

confidence: 98%

“…To promote root induction, healthy microshoots at least 2 cm in length were cultured in DKW-C medium containing one-half macronutrients, 117 mM sucrose, and 50 lM IBA (Vahdati et al 2004;Dolcet-Sanjuan et al 2004). IBA gave results superior to indole-3-acetic acid (IAA) or 1-naphthaleneacetic acid (NAA).…”

Section: Culture Conditionsmentioning

confidence: 99%

Improved walnut mass micropropagation through the combined use of phloroglucinol and FeEDDHA

Licea-Moreno

Contreras

Morales

et al. 2015

Plant Cell Tiss Organ Cult

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Despite the socioeconomic importance of walnut trees, poor rooting and recalcitrance to in vitro culture have hampered the establishment of high-yield clonal plantations. To improve walnut micropropagation, we introduced several modifications to current methods and evaluated the effects on microshoot performance and acclimatization. Nine selected genotypes (13-year-old trees) of the commercial hybrid Juglans major 209 9 J. regia were cultured in vitro on DKW-C medium supplemented with 4.4 lM BA and 50 lM IBA. A protocol was developed that relies on the sequential use of 0.4 and 0.2 mM phloroglucinol during shoot multiplication, but not at later stages, so as to maximize shoot growth without inhibiting root formation. Moreover, substituting FeEDTA by FeEDDHA diminished chlorotic symptoms and significantly improved the rooting ability of all genotypes, with up to 90 % microshoots developing viable roots at 6.81 mg/L Fe 3? . The addition of 83.2 mM glucose during the root expression phase was particularly efficient at promoting plant survival during acclimatization, compared to equimolar amounts of the alternative sugars sucrose and fructose. At the proposed working concentrations, the aforementioned compounds did not cause any deleterious effects on the nine genotypes studied. Microscopic analysis revealed the physical continuity between adventitious roots and stem cambial tissue. Analysis of leaf genomic DNA with eight polymorphic microsatellite markers was supportive of the clonal fidelity and genetic stability of the micropropagated material. Successful clonal plantations (over 5800 ramets) have been established by applying this protocol.

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“…Progresses in micropropagation of walnut have been achieved using single node and shoot tip cultures (Vahdati et al 2004). Somatic embryos are important not only as a target tissue for genetic transformation but also as a tool for mass clonal propagation (McGranahan et al 1990).…”

Section: Introductionmentioning

confidence: 99%

Effect of exogenous ABA on somatic embryo maturation and germination in Persian walnut (Juglans regia L.)

Vahdati

Bayat

Ebrahimzadeh

et al. 2008

Plant Cell Tiss Organ Cult

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Low efficiency of embryo maturation, germination and conversion to plantlets is a major problem in many species including Persian walnut. We studied the effects of abscisic acid (ABA) and sucrose, on the maturation and germination of Persian walnut (Juglans regia) somatic embryos. Individual globular somatic embryos were grown on a maturation medium supplemented with different combinations of ABA and sucrose for ca. 1 month, until shoot meristems and radicles had developed. White and opaque embryos in late cotyledonary stage were subjected to desiccation after the culture period on maturation media. The number of germinated somatic embryos was influenced by the concentrations of ABA in the maturation medium. The best treatment for germination, in which both shoot and root were developed contained 2 mg l -1 ABA and resulted in 41% conversion of embryos into plantlets. Regeneration was reduced at higher levels of ABA. While ABA always reduced the rate of secondary embryogenesis, treatments containing 4.0% sucrose significantly increased the number of secondary embryos. On the other hand, sucrose had little influence on maturation. Normal and abnormal embryos were verified anatomically.

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Rooting and Acclimatization of In Vitro-grown Shoots from Mature Trees of Three Persian Walnut Cultivars

Cited by 47 publications

References 13 publications

Evaluation of red fluorescent protein (DsRed) as alternative visual marker of genetic transformation in cassava (Manihot esculenta Crantz)

Evaluation of red fluorescent protein (DsRed) as alternative visual marker of genetic transformation in cassava (Manihot esculenta Crantz)

Improved walnut mass micropropagation through the combined use of phloroglucinol and FeEDDHA

Effect of exogenous ABA on somatic embryo maturation and germination in Persian walnut (Juglans regia L.)

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