Plant Regeneration from Callus Derived from Immature Embryo Cotyledons of Prunus mume

in HortScience
Authors: G.G. Ning1 and M.Z. Bao1
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  • 1 Key Laboratory of Horticultural Plant Biology, Ministry of Education, College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan 430070, P.R. China

A regeneration protocol for Prunus mume sieb.et Zucc was developed through indirect organogenesis. Immature cotyledons were excised from the open-pollinated seeds of two cultivars and cultured on a modified MS medium supplemented with various combinations of plant growth regulators. Shoot-organogenic calli were induced on half-strength MS medium supplemented either with combinations of 2.2 μm benzyladenine (BA), 5.4 to 10.8 μm 1-naphthaleneacetic acid (NAA), and 0 to 5.0 μm indolebutyric acid (IBA) or with combinations of 2.2 μm BA, 4.5 to 9.0 μm dichlophenoxyacetic acid (2,4-D), and 0 to 5.0 μm IBA. High frequencies of shoot regeneration (81.5% and 91.3% in P. mume cvs. Lv'e and Xuemei, respectively) were obtained from smooth-white nodular calli cultured on a half-strength MS medium supplemented with 2.2 μm BA, 2.2 μM thidiazuron, and 1.0 μm IBA. A high rate of rooting (90.2% and 88.9% in ‘Lv'e’ and ‘Xuemei’, respectively) occurred when shoots were cultured on WPM supplemented with 5.0 μm IBA. Chemical names used: benzyladenine (BA), thidiazuron (TDZ), indolebutyric acid (IBA), dichlophenoxyacetic acid (2,4-D), 1-naphthaleneacetic acid (NAA).

Abstract

A regeneration protocol for Prunus mume sieb.et Zucc was developed through indirect organogenesis. Immature cotyledons were excised from the open-pollinated seeds of two cultivars and cultured on a modified MS medium supplemented with various combinations of plant growth regulators. Shoot-organogenic calli were induced on half-strength MS medium supplemented either with combinations of 2.2 μm benzyladenine (BA), 5.4 to 10.8 μm 1-naphthaleneacetic acid (NAA), and 0 to 5.0 μm indolebutyric acid (IBA) or with combinations of 2.2 μm BA, 4.5 to 9.0 μm dichlophenoxyacetic acid (2,4-D), and 0 to 5.0 μm IBA. High frequencies of shoot regeneration (81.5% and 91.3% in P. mume cvs. Lv'e and Xuemei, respectively) were obtained from smooth-white nodular calli cultured on a half-strength MS medium supplemented with 2.2 μm BA, 2.2 μM thidiazuron, and 1.0 μm IBA. A high rate of rooting (90.2% and 88.9% in ‘Lv'e’ and ‘Xuemei’, respectively) occurred when shoots were cultured on WPM supplemented with 5.0 μm IBA. Chemical names used: benzyladenine (BA), thidiazuron (TDZ), indolebutyric acid (IBA), dichlophenoxyacetic acid (2,4-D), 1-naphthaleneacetic acid (NAA).

Prunus mume is a beautiful deciduous tree with fragrant flowers that blooms very early in the spring. It is renowned for its attractive blossoms and longevity throughout China and Japan. In addition, P. mume has medical value (Dogasaki et al., 2004) with “pickled plums,” made from the unripe fruits of P. mume, being used to treat vomiting and fever (Chen, 1962). As a result of its economic importance as an ornamental and medicinal tree, there is significant interest in breeding elite varieties of P. mume, and this has primarily focused on using molecular markers to identify desirable phenotypes and characterizing the mechanism of self-incompatibility (Entani et al., 2003; Tao et al., 2000).

During the last 2 decades, efficient in vitro culture systems have been developed for various species of the Prunus genus. The majority of these have been achieved by inducing regeneration from either immature tissues (Hashmi et al., 1997; Hokanson and Pooler, 2000; Mante et al., 1989; Pooler and Scorza, 1995; Tang et al., 2000) or mature explants (Andrea and Johannes, 2005; Bhagwat and David, 2004; Cheong and Pooler, 2004; Declerck and Korban, 1996; Gentile et al., 2002; Hammatt and Grant,.1998; Neil and Neil, 2000; Olaya et al., 2000). For P. mume, an in vitro culture system has been reported for the induction of callus, but plant regeneration was not achieved (Liu and Chen, 1999). Although it is possible to conduct traditional breeding programs in P. mume, particularly with the assistance of molecular markers, this is a protracted process because of the long intergeneration times. Therefore, to accelerate the genetic improvement of P. mume, genetic engineering appears to be an attractive option allowing the introduction of heterologous genes for the enhancement of ornamental or other valued characteristics. Efficient plant regeneration from in vitro cultures is essential for the genetic transformation of P. mume. However, to date, there are very few reports of the successful regeneration of plants from cultures of P. mume.

This article presents the first report of the successful regeneration of plants of P. mume achieved through indirect organogenesis from cotyledons of immature embryos. In the course of this work, we also investigated the effects of the original explant source and the light environment on the rate of shoot organogenesis from callus cultures.

Materials and Methods

Plant material.

Open-pollinated, immature seeds (≈40 to 50 d after pollination) were collected from two mature trees of P. mume cultivars, Xuemei and Lv'e. The trees had been growing in the Wuhan Mei flower garden (Wuhan, P.R. China) for over 10 years at the time of seed collection during Apr. and May 2005. The seeds were stored at 4 °C until use. After being washed thoroughly with running tap water for 30 min, the immature seeds were disinfected by immersion in 70% (v/v) ethanol for 30 to 60 s followed by one wash in sterile water, then immersion and agitation in a 0.1% (w/v) aqueous solution of HgCl2 for 20 min, followed by three washes in sterile water. Cotyledons were aseptically excised and placed horizontally on the medium for callus induction. To investigate the effect of the original explant source on callus induction and subsequent plant regeneration, individual cotyledons were divided into proximal or distal sections with the embryonic axis removed.

Culture initiation and plant regeneration.

The basal medium consisted of half-strength MS medium (Murashige and Skoog, 1962) supplemented with 3% (w/v) sucrose and 0.8% (w/v) agar (Sigma A1296, Sigma, St. Louis). This was supplemented with various combinations of plant growth regulators (see subsequently). The pH was adjusted to 5.8 with 1 m NaOH before autoclaving at 121 °C for 20 to 25 min. The media were dispensed into 90 mm petri dishes (25 mL of medium per dish), in the case of callus induction, or plastic boxes (250 mL, made in China; 40 mL per box) for shoot induction, shoot elongation, and rooting procedures.

Experiments of callus induction from cotyledon explants were performed on half-strength MS media supplemented with various levels of benzyladenine (BA), dichlophenoxyacetic acid (2,4-D), 1-naphthaleneacetic acid (NAA), and indolebutyric acid (IBA) supplied in 11 combinations (see Table 1). To stimulate adventitious shoot induction, callus outgrowths were transferred to various media supplemented with various levels of either BA and NAA or thidiazuron (TDZ) and IBA (Table 2).

Table 1.

Response of callus induction to plant growth regulators in immature Prunus mume cotyledons (cultivars Xuemei and Lv'e) on half-strength MS medium.z

Table 1.
Table 2.

Percentage of regeneration from cotyledon-derived callus in relation to plant growth regulators used with half-strength MS medium for two Prunus mume cultivars (Xuemei and Lv'e).z

Table 2.

All the cultures (unless stated otherwise) were maintained in a growth chamber at 25 ± 2 °C under a 14-h photoperiod of 50 μmol·m−2·s−1 of photosynthetic photon flux (PPF).

Effect of callus source and light on shoot induction.

It was found that half-strength MS medium supplemented with 2.2 μm BA, 10.0 μm NAA, and 5.0 μm IBA produced the most vigorous callus growth, and half-strength MS medium supplemented with 2.2 μm BA, 2.2 μm TDZ, and 1.0 μm IBA yielded the highest number of shoots. Therefore, these two media were used in all further experiments designed to test the effects of callus source and light conditions on rates of plant regeneration.

Shoot elongation, rooting, and acclimatization.

To promote elongation growth of shoots, the regenerated buds were cultured on WPM medium (Lloyd and McCown, 1980) supplemented with 2.2 μm BA, 0.45 μm TDZ, and 1.0 μm IBA. After incubation for 3 weeks, the most vigorous shoots were cut at the base and transferred to a WPM medium supplemented with 2.5 or 5.0 μm IBA and incubated under lights to induce rooting. After 6 weeks, rooted plantlets (defined as having at least one root greater than 10 mm in length) were washed thoroughly with running tap water, to remove the culture medium, and transplanted into plastic pots containing a mixture of 2 peatmoss : 2 garden soil : 1 sand (by volume). These were then kept in a growth chamber set to 70% relative humidity and 19 °C. After 2 weeks, the pots were transferred to a greenhouse and maintained at average 19 °C for further development.

Statistical analysis.

For callus induction, seven explants were incubated under each treatment regimen. Experiments of callus initiation and shoot induction were repeated at least three times. The differences between mean values (obtained from at least two independent experiments) were statistically evaluated by analysis of variance (ANOVA) using Duncan's multiple-range test with the least significant difference set to 5% probability.

Results

Callus initiation.

No callus was obtained when immature cotyledons of P. mume (Fig. 1A) were cultured on a medium that was free from plant growth regulators or contained less than 2.7 μm NAA or 2.2 or less BA. However, calli (Fig. 1B) could be obtained when explants were cultured on half-strength MS supplemented with different concentrations of either BA, NAA, and IBA or BA, 2,4-D, and IBA (Tables 1 and 2). There were significant differences (P < 0.05) in the percentage of callus induction resulting from incubation of cotyledon explants on the various media. Across the combinations of plant growth hormones tested and the two cultivars of P. mume, the percentage of callus formation varied from 22% to 97% (Table 1). Growth of callus that was induced on various media varied considerably with respect to physical appearance and could be classified into three general types, i.e., 1 = light yellow, loose and globular; 2 = smooth white, loose and nodular (Fig. 1B); or 3 = light yellow, hard and globular.

Fig. 1.
Fig. 1.

Plant regeneration through shoot organogenesis from callus derived from cotyledons of immature embryos of Prunus mume: (A) Cotyledons from P. mume immature embryos. (B) Calli formed from the cotyledon explants (arrow). (C) Shoots regenerated from smooth white nodular calli of cotyledon explants in darkness. (D) Shoots regenerated from smooth-white nodular calli under light. (E) Shoot elongation. (F) Rooting (view of the bottom of plastic box).

Citation: HortScience horts 42, 3; 10.21273/HORTSCI.42.3.744

Shoot regeneration from callus.

In the most part, shoots regenerated through organogenesis were derived from the type 2 form of callus (see previously) (Fig. 1B) that had either originated from cotyledons or were from primary callus cultures. These calli maintained their morphogenetic potential for at least two rounds of subculturing. When these calli were cultured on half-strength MS supplemented with 2.7 μm NAA and 13.2 to 22.0 μm BA, small regenerated shoots (Fig. 1C, D) were observed to form on the callus after 4 weeks, but this tended to occur at fairly low frequencies. The further incorporation of TDZ in the regeneration medium significantly stimulated shoot regeneration. Thus, a medium supplemented with 2.2 μm BA and 1.0 μm IBA plus 2.25 μm TDZ produced the maximal regeneration frequencies of 81.5% and 91.3% in the ‘Lv'e’ and ‘Xuemei’ cultivars, respectively (Table 2). No significant difference in regeneration frequency was detected between calli initiated from the proximal or distal sections of the cotyledons for either cultivar (Table 3). By contrast, the ambient light environment produced different regeneration responses in the two cultivars (Table 4). In ‘Lv'e’, regeneration frequency did not differ significantly under light compared with dark conditions as tested by ANOVA analysis (P < 0.05). In ‘Xuemei’; however, it was apparent that continuous darkness was beneficial for regeneration (Table 4).

Table 3.

Effect of explant source (proximal or distal to the embryonic axis) on adventitious shoot formation of two cultivars (Xuemei and Lv'e) of Prunus mume.z

Table 3.
Table 4.

Effect of light on adventitious shoot formation from cotyledon explants of the Xuemei and Lv'e cultivars of Prunus mume.z

Table 4.

Shoot elongation, rooting, and acclimatization.

After growth in elongation medium for ≈3 weeks, shoots at least 1 cm in length (Fig. 1E) were cut and transferred to rooting medium. Within 4 weeks, these shoots had produced roots from the base of the excision site (Fig. 1F). The number of shoots that produced one or more roots (according to 50 tested shoots) and the number of roots (greater than 5 mm in length) per shoot differed significantly when cultured on WPM media as supplemented with 2.5 or 5.0 μm IBA (Table 5). By contrast, there was no significant difference between the two cultivars at a given concentration of IBA. After 6 weeks of rooting initiation, the plantlets were transferred to soil for “hardening.” The survival rate of these regenerated plantlet was greater than 90% under greenhouse conditions.

Table 5.

Rooting of Prunus. mume Xuemei and Lv'e after 4 weeks of culture on different media.z

Table 5.

Discussion

This is the first report, to our knowledge, of the successful in vitro regeneration of P. mume through organogenesis from callus derived from immature cotyledons.

In Prunus species, somatic embryogenesis has been the most commonly reported regeneration pathway (Cheong and Pooler, 2004; Hashmi et al., 1997; Tang et al., 2000). Directive shoot differentiation had been reported from immature cotyledons of almond (Phillip et al., 2001; Saafi and Dulal, 2002) and some other woody plant cultures (Tan and Furtek, 2003; Wei et al., 2005). In this study of Prunus mume, it was possible to obtain abundant callus from immature embryos cultured on a medium supplemented with specific concentrations of plant growth regulators (Table 1), and shoots could be readily regenerated from the smooth-white nodular form of callus. This result is consistent with that reported for peach (Hammerschlag et al., 1985; Hashmi et al., 1997). During callus induction, increasing the concentration of NAA (in a medium also containing 2.2 μm BA) increased the percentage of callus formation (Table 1) and furthermore eliminated the secretion of phenolic substances from the cut surface of the cotyledons. This observation is consistent with Olaya et al. (2000) who reported that such phenolic substances are particularly liable to be oxidized by auxin oxidase when there is a high concentration of NAA in the culture medium. Callus was also readily obtained on media supplemented with 2.2 μm BA in combination with either of two auxins, i.e., NAA or 2,4-D and IBA. A high level of 2,4-D (9.0 μm) in combination with a low level of BA and IBA (2.2 and 1.0 μm, respectively) led to high callus yields. This was contrary to reports in other plant species (Luo et al., 1999; Nolan et al., 1989). Previous reports of other Prunus species have indicated that plantlet regeneration from leaves shows that WPM basic medium is more suitable for the induction of adventitious shoots than is MS basal medium (Andrea and Johannes, 2005; Bhagwat and David, 2004; Hammatt and Grant, 1998; Neil and Neil, 2000; Tang et al., 2002). However, a high frequency regeneration of P. mume was obtained from organogenic callus derived from cotyledons that had been cultured on half-strength MS media supplemented with 2.2 μm BA, 2.25 μm TDZ, and 1.0 μm IBA. There was no difference in regeneration frequency from those calli derived from the distal, as opposed to proximal, parts of the cotyledons (Table 3). This is in contrast to observations from other Prunus species (Mante et al., 1989; Tang et al., 2000).

With regard to the frequency of shoot organogenesis from calli derived from the ‘Xuemei’ cultivar, the effect of a light versus dark incubation was significant (Table 4). In ‘Lv'e’, there was no significant effect. This result indicates that the importance of the light environment on shoot organogenesis from cotyledon-derived calli is genotype-dependent.

Recently, successful genetic transformation has been reported for a number of plant species through the use of cotyledon explants (Choi et al., 2001, Zaragoz et al., 2004), and transgenic plants have been obtained for some other Prunus species (Rosa et al., 2004). Here, we have described the culture conditions that can produce an efficient frequency of plantlet regeneration from immature cotyledon explants of Prunus mume. The regeneration method presented here is an essential basis for the introduction of heterologous genes through Agrobacterium-mediated transformation for the genetic improvement of this important tree species.

Literature Cited

  • Andrea, M. & Johannes, A.J. 2005 In vitro plant regeneration from leaves and internode sections of sweet cherry cultivars (Prunus avium. L) Plant Cell Rep. 24 468 476

    • Search Google Scholar
    • Export Citation
  • Bhagwat, B. & David, W.L. 2004 In vitro shoot regeneration from leaves of sweet cherry (Prunus avium) ‘Lapins’ and ‘Sweetheart’ Plant Cell Tissue Organ Cult. 78 173 181

    • Search Google Scholar
    • Export Citation
  • Chen, J.Y. 1962 Studies of Chinese Mei (Prunus mume Sieb. et Zucc)—The origin and cultivation history Acta Horticulturae Sinica. 1 69 78

  • Cheong, E.J. & Pooler, M.R. 2004 Factors affecting somatic embryogenesis in Prunus incisa cv. February Pink Plant Cell Rep. 22 810 815

  • Choi, Y.E., Yang, D.C., Kusano, T. & Sano, H. 2001 Rapid and efficient Agrobacterium mediated transformation of Panax ginseng by plasmolyzing pre-treatment of cotyledons Plant Cell Rep. 20 616 621

    • Search Google Scholar
    • Export Citation
  • Declerck, V. & Korban, S.S. 1996 Influence of growth regulators and carbon sources on callus induction, growth and morphogenesis from leaf tissues of peach (Prunus persica L. Batsch) HortScience 71 49 55

    • Search Google Scholar
    • Export Citation
  • Dogasaki, C., Kakuno, Y., Honda, M., Takada, N., Maruyama, T., Nishijima, M., Adachi, Y., Ohno, N., Yadomae, T. & Miyazaki, T. 2004 Contribution to Immunochemical Analysis of Polysaccharides in Medicinal Plants Drug Design Reviews 1 153 159

    • Search Google Scholar
    • Export Citation
  • Entani, T., Iwano, M., Shiba, H., Che, F.S., Isogai, A. & Takayama, S. 2003 Comparative analysis of the self-incompatibility (S-) locus region of Prunus mume: Identification of a pollen-expressed F-box gene with allelic diversity Genes Cells 8 203 213

    • Search Google Scholar
    • Export Citation
  • Gentile, A., Monticelli, S. & Damiano, C. 2002 Adventitious shoot regeneration in peach [Prunus persica (L.) Batsch] Plant Cell Rep. 20 1011 1016

  • Hammatt, N. & Grant, N.J. 1998 Shoot regeneration from leaves of Prunus serotina Ehrh. (black cherry) and P. avium L. (wild cherry) Plant Cell Rep. 17 526 530

    • Search Google Scholar
    • Export Citation
  • Hammerschlag, F.A., Bauchan, G. & Scorza, R. 1985 Regeneration of peach plants from callus derived from immature embryos Theor. Appl. Genet. 70 248 251

    • Search Google Scholar
    • Export Citation
  • Hashmi, G., Huettel, R., Meyer, R., Krusberg, L. & Hammerschlag, F. 1997 RAPD analysis of somaclonal variants derived from embryo callus cultures of peach Plant Cell Rep. 16 624 627

    • Search Google Scholar
    • Export Citation
  • Hokanson, K.E. & Pooler, M.R. 2000 Regeneration of ornamental cherry (Prunus) taxa from mature stored seed HortScience 35 745 748

  • Liu, Q.L. & Chen, Q.H. 1999 Preliminary reports of callus culture on Prunus mume Journal of Beijing Forestry University. 21 400 405

  • Lloyd, G.B. & McCown, B.H. 1980 Commercially feasible micropropagation of mountain laurel, Kalmia latifolia, by use of shoot-tip culture Proc Int Plant Prop Soc. 30 421 427

    • Search Google Scholar
    • Export Citation
  • Luo, J.P., Jia, J.F., Gu, Y.H. & Liu, J. 1999 High frequency somatic embryogenesis and plant regeneration in callus cultures of Astragalus adsurgens. Pall Plant Sci. 143 93 99

    • Search Google Scholar
    • Export Citation
  • Mante, S., Scorza, R. & Cordts, J.M. 1989 Plant regeneration from cotyledons of Prunus persica, Prunus domestica, and Prunus cerasus Plant Cell Tissue Organ Cult. 19 1 11

    • Search Google Scholar
    • Export Citation
  • Murashige, T. & Skoog, F. 1962 A revised media for rapid growth and bioassays with tobacco tissue cultures Physiol. Plant. 15 473 497

  • Neil, J.G. & Neil, H. 2000 Adventitious shoot development from wild cherry (Prunus avium L.) leaves New For. 20 287 295

  • Nolan, K.E., Rose, R.J. & Gost, J.R. 1989 Regeneration of Medicago truncatula from tissue culture: Increased somatic embryogenesis using explants from regenerated plant Plant Cell Rep. 8 279 281

    • Search Google Scholar
    • Export Citation
  • Olaya, P.T., Jose, E., Alicia, V. & Lorenzo, B. 2000 Assessment of factors affecting adventitious shoot regeneration from in vitro cultured leaves of apricot Plant Sci. 158 61 70

    • Search Google Scholar
    • Export Citation
  • Phillip, J.A., Freddi, A.H., Terry, B., Graham, G.C. & Margaret, S. 2001 Regeneration of almond from immature seed cotyledons Plant Cell Tissue Organ Cult. 67 221 226

    • Search Google Scholar
    • Export Citation
  • Pooler, M.R. & Scorza, R. 1995 Regeneration of peach [Prunus persica (L.) Batsch] rootstock cultivars from cotyledons of mature stored seed HortScience 30 355 356

    • Search Google Scholar
    • Export Citation
  • Rosa, M.P.C., Amparo, P.S., Lorenzo, G.F., José-Pío, B. & Luis, A.C. 2004 Transgenic peach plants (Prunus persica L.) produced by genetic transformation of embryo sections using the green fluorescent protein (GFP) as an in vivo marker Mol. Breed. 14 419 427

    • Search Google Scholar
    • Export Citation
  • Saafi, H. & Dulal, B. 2002 In vitro plantlet regeneration from cotyledons of the tree-legume Leucaena leucocephala Plant Growth Regul. 38 279 285

  • Tan, C.L. & Furtek, D.B. 2003 Development of an in vitro regeneration system for Theobroma cacao from mature tissues Plant Sci. 164 407 412

  • Tang, H., Ren, Z.L. & Gabi, K. 2000 Somatic embryogenesis and organogenesis from immature embryo cotyledons of three sour cherry cultivars (Prunus cerasus L.) Sci. Hort. 83 109 126

    • Search Google Scholar
    • Export Citation
  • Tang, H., Ren, Z.L., Reustle, G. & Krczal, G. 2002 Plant regeneration from leaves of sweet and sour cherry cultivars Sci. Hort. 93 235 244

  • Tao, R., Habu, T. & Yamane, H. 2000 Molecular markers for self-compatibility in Japanese apricot (Prunus mume) HortScience 35 121 123

  • Wei, T. & Newton, R.J. 2005 Plant regeneration from callus cultures derived from mature zygotic embryos in white pine (Pinus strobus L.) Plant Cell Rep. 24 1 9

    • Search Google Scholar
    • Export Citation
  • Zaragoz, C., Mu, J. & Bertomeu, A. 2004 Regeneration of herbicide-tolerant black locust transgenic plants by SAAT Plant Cell Rep. 22 832 838

Contributor Notes

We thank all present and past colleagues of our laboratory for constructive discussion and technical support. We are also grateful to all the staff of the Wuhan P. mume garden, China, for providing experimental materials and Alex McCormac of England for critical reading and editing of the manuscript.

To whom reprint requests should be addressed; e-mail whnobleman2004@yahoo.com.cn

  • View in gallery

    Plant regeneration through shoot organogenesis from callus derived from cotyledons of immature embryos of Prunus mume: (A) Cotyledons from P. mume immature embryos. (B) Calli formed from the cotyledon explants (arrow). (C) Shoots regenerated from smooth white nodular calli of cotyledon explants in darkness. (D) Shoots regenerated from smooth-white nodular calli under light. (E) Shoot elongation. (F) Rooting (view of the bottom of plastic box).

  • Andrea, M. & Johannes, A.J. 2005 In vitro plant regeneration from leaves and internode sections of sweet cherry cultivars (Prunus avium. L) Plant Cell Rep. 24 468 476

    • Search Google Scholar
    • Export Citation
  • Bhagwat, B. & David, W.L. 2004 In vitro shoot regeneration from leaves of sweet cherry (Prunus avium) ‘Lapins’ and ‘Sweetheart’ Plant Cell Tissue Organ Cult. 78 173 181

    • Search Google Scholar
    • Export Citation
  • Chen, J.Y. 1962 Studies of Chinese Mei (Prunus mume Sieb. et Zucc)—The origin and cultivation history Acta Horticulturae Sinica. 1 69 78

  • Cheong, E.J. & Pooler, M.R. 2004 Factors affecting somatic embryogenesis in Prunus incisa cv. February Pink Plant Cell Rep. 22 810 815

  • Choi, Y.E., Yang, D.C., Kusano, T. & Sano, H. 2001 Rapid and efficient Agrobacterium mediated transformation of Panax ginseng by plasmolyzing pre-treatment of cotyledons Plant Cell Rep. 20 616 621

    • Search Google Scholar
    • Export Citation
  • Declerck, V. & Korban, S.S. 1996 Influence of growth regulators and carbon sources on callus induction, growth and morphogenesis from leaf tissues of peach (Prunus persica L. Batsch) HortScience 71 49 55

    • Search Google Scholar
    • Export Citation
  • Dogasaki, C., Kakuno, Y., Honda, M., Takada, N., Maruyama, T., Nishijima, M., Adachi, Y., Ohno, N., Yadomae, T. & Miyazaki, T. 2004 Contribution to Immunochemical Analysis of Polysaccharides in Medicinal Plants Drug Design Reviews 1 153 159

    • Search Google Scholar
    • Export Citation
  • Entani, T., Iwano, M., Shiba, H., Che, F.S., Isogai, A. & Takayama, S. 2003 Comparative analysis of the self-incompatibility (S-) locus region of Prunus mume: Identification of a pollen-expressed F-box gene with allelic diversity Genes Cells 8 203 213

    • Search Google Scholar
    • Export Citation
  • Gentile, A., Monticelli, S. & Damiano, C. 2002 Adventitious shoot regeneration in peach [Prunus persica (L.) Batsch] Plant Cell Rep. 20 1011 1016

  • Hammatt, N. & Grant, N.J. 1998 Shoot regeneration from leaves of Prunus serotina Ehrh. (black cherry) and P. avium L. (wild cherry) Plant Cell Rep. 17 526 530

    • Search Google Scholar
    • Export Citation
  • Hammerschlag, F.A., Bauchan, G. & Scorza, R. 1985 Regeneration of peach plants from callus derived from immature embryos Theor. Appl. Genet. 70 248 251

    • Search Google Scholar
    • Export Citation
  • Hashmi, G., Huettel, R., Meyer, R., Krusberg, L. & Hammerschlag, F. 1997 RAPD analysis of somaclonal variants derived from embryo callus cultures of peach Plant Cell Rep. 16 624 627

    • Search Google Scholar
    • Export Citation
  • Hokanson, K.E. & Pooler, M.R. 2000 Regeneration of ornamental cherry (Prunus) taxa from mature stored seed HortScience 35 745 748

  • Liu, Q.L. & Chen, Q.H. 1999 Preliminary reports of callus culture on Prunus mume Journal of Beijing Forestry University. 21 400 405

  • Lloyd, G.B. & McCown, B.H. 1980 Commercially feasible micropropagation of mountain laurel, Kalmia latifolia, by use of shoot-tip culture Proc Int Plant Prop Soc. 30 421 427

    • Search Google Scholar
    • Export Citation
  • Luo, J.P., Jia, J.F., Gu, Y.H. & Liu, J. 1999 High frequency somatic embryogenesis and plant regeneration in callus cultures of Astragalus adsurgens. Pall Plant Sci. 143 93 99

    • Search Google Scholar
    • Export Citation
  • Mante, S., Scorza, R. & Cordts, J.M. 1989 Plant regeneration from cotyledons of Prunus persica, Prunus domestica, and Prunus cerasus Plant Cell Tissue Organ Cult. 19 1 11

    • Search Google Scholar
    • Export Citation
  • Murashige, T. & Skoog, F. 1962 A revised media for rapid growth and bioassays with tobacco tissue cultures Physiol. Plant. 15 473 497

  • Neil, J.G. & Neil, H. 2000 Adventitious shoot development from wild cherry (Prunus avium L.) leaves New For. 20 287 295

  • Nolan, K.E., Rose, R.J. & Gost, J.R. 1989 Regeneration of Medicago truncatula from tissue culture: Increased somatic embryogenesis using explants from regenerated plant Plant Cell Rep. 8 279 281

    • Search Google Scholar
    • Export Citation
  • Olaya, P.T., Jose, E., Alicia, V. & Lorenzo, B. 2000 Assessment of factors affecting adventitious shoot regeneration from in vitro cultured leaves of apricot Plant Sci. 158 61 70

    • Search Google Scholar
    • Export Citation
  • Phillip, J.A., Freddi, A.H., Terry, B., Graham, G.C. & Margaret, S. 2001 Regeneration of almond from immature seed cotyledons Plant Cell Tissue Organ Cult. 67 221 226

    • Search Google Scholar
    • Export Citation
  • Pooler, M.R. & Scorza, R. 1995 Regeneration of peach [Prunus persica (L.) Batsch] rootstock cultivars from cotyledons of mature stored seed HortScience 30 355 356

    • Search Google Scholar
    • Export Citation
  • Rosa, M.P.C., Amparo, P.S., Lorenzo, G.F., José-Pío, B. & Luis, A.C. 2004 Transgenic peach plants (Prunus persica L.) produced by genetic transformation of embryo sections using the green fluorescent protein (GFP) as an in vivo marker Mol. Breed. 14 419 427

    • Search Google Scholar
    • Export Citation
  • Saafi, H. & Dulal, B. 2002 In vitro plantlet regeneration from cotyledons of the tree-legume Leucaena leucocephala Plant Growth Regul. 38 279 285

  • Tan, C.L. & Furtek, D.B. 2003 Development of an in vitro regeneration system for Theobroma cacao from mature tissues Plant Sci. 164 407 412

  • Tang, H., Ren, Z.L. & Gabi, K. 2000 Somatic embryogenesis and organogenesis from immature embryo cotyledons of three sour cherry cultivars (Prunus cerasus L.) Sci. Hort. 83 109 126

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  • Tang, H., Ren, Z.L., Reustle, G. & Krczal, G. 2002 Plant regeneration from leaves of sweet and sour cherry cultivars Sci. Hort. 93 235 244

  • Tao, R., Habu, T. & Yamane, H. 2000 Molecular markers for self-compatibility in Japanese apricot (Prunus mume) HortScience 35 121 123

  • Wei, T. & Newton, R.J. 2005 Plant regeneration from callus cultures derived from mature zygotic embryos in white pine (Pinus strobus L.) Plant Cell Rep. 24 1 9

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  • Zaragoz, C., Mu, J. & Bertomeu, A. 2004 Regeneration of herbicide-tolerant black locust transgenic plants by SAAT Plant Cell Rep. 22 832 838

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