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Somatic embryogenesis, plant regeneration, and cryopreservation for Torreya taxifolia, a highly endangered coniferous species
Abstract
Torreya taxifolia Arn., an ancient evergreen tree, is on the brink of extinction from attack by a fungal disease, recently reported to be caused by a novel isolate of Fusarium. We report the development of a somatic embryogenesis tissue culture system that can be used for cryogenic storage of T. taxifolia cultures and subsequent plant regeneration. Initiation of embryogenic tissue from immature zygotic embryos occurred on a conifer tissue culture medium containing 0.25 % activated charcoal, 43.8 mM maltose, 0.5 mM 2,4-dichlorophenoxacetic acid, 0.2 mM 6-benzylaminopurine, 0.2 mM kinetin, 0.1 μM brassinolide, 3.8 μM abscisic acid, 20.5 μM biotin, 1.13 μM folic acid, 1.28 mM 2(n-morpholino)ethanesulfonic acid and 0.69 mM pyruvic acid. Embryo induction ranged from 60 to 100 % across six seed sources. Somatic embryo development occurred on a medium containing 43.8 mM maltose, 1 % activated charcoal, 37.8 μM abscisic acid, 20.5 μM biotin, 0.1 μM brassinolide, 0.205 mM folic acid, 1.28 mM 2(n-morpholino)ethanesulfonic acid and 0.69 mM pyruvic acid. Germination of somatic embryos ranged from 64 to 82 %. Embryogenic tissue cultures from 30 genotypes representing seed from six mother trees were cryopreserved, and culture recovery was demonstrated after freezing. In contrast to many other coniferous tree seeds, the measured water potential (−MPa) of T. taxifolia megagametophyte tissue rose greatly during seed after-ripening. Duplication of this rise in vitro allowed development of somatic embryos to the cotyledonary stage.
  • References (34)
  • Cited By (18)
Michael R. Becwar2
Estimated H-index: 2
,
Gerald S. Pullman1
Estimated H-index: 1
Somatic embryogenesis, the formation and development of embryos from somatic (vegetative) tissues under in vitro conditions, was first discovered in carrot (Steward et al., 1958) and in conifers with Picea abies in 1985 (Hakman et al., 1985). The first report of somatic embryogenesis with Pinus taeda was in 1987 (Gupta & Durzan, 1987).
Ref 36Cited 84
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Claudio Stasolla26
Estimated H-index: 26
(University of Winnipeg),
Edward C. Yeung39
Estimated H-index: 39
(University of Calgary)
Somatic embryogenesis of coniferous species was first reported more than 20 years ago. Since then, there has been an explosion of research aimed at developing and optimizing protocols for efficient regeneration of plantlets. Although routinely used both as a means of propagation, as well as a valuable model system for investigating the structural, physiological, and molecular events occurring during embryo development, in vitro embryogenesis is still problematic for some coniferous species. Majo...
Ref 104Cited 182 Source Cite this paper
P. K. Gupta1
Estimated H-index: 1
(University of California),
D. J. Durzan32
Estimated H-index: 32
(University of California)
Loblolly pine (Pinus taeda L.) is the leading commercial timber species in the southern United States. Carolus Linnaeus gave loblolly pine its scientific name, Pinus taeda, over 225 years ago. Taeda is the ancient name for resinous pines. It comes from the Latin, meaning torch. In Roman times, taeda was applied to several of the hard, pitchy pines (Wahlenberg 1960).
Ref 23Cited 100
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Nanfei Xu6
Estimated H-index: 6
,
Barbara Johns4
Estimated H-index: 4
,
Gerald S. Pullman19
Estimated H-index: 19
... more
Performing RNA differential display analysis on small tissue samples is difficult since much RNA, the initial template for the reaction, is lost during conventional isolation procedures. We have developed a rapid method which employs oligo-dT beads to capture mRNA from cell lysates. Subsequent reactions are primed directly from the beads, thus RT and PCR reactions can be completed within a few hours of tissue harvest. This approach allows us to perform differential display on a single pine embry...
Ref 15Cited 23
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Aljos Farjon1
Estimated H-index: 1
Foreword Preface The conifers of the world, an introduction The distribution and ecology of conifers The economic importance of conifers The conservation of conifer diversity Synopsis of families and genera Taxonomic treatment of families, with keys to families Taxonomic treatment of genera and species (in alphabetical order), with keys to sections, subsections and species Glossary References Lists of illustrations Index to botanical names of conifers
Cited 102
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Gerald S. Pullman19
Estimated H-index: 19
,
Shannon Johnson8
Estimated H-index: 8
,
Grabner Peter1
Estimated H-index: 1
... more
Clonal production of loblolly pine (Pinus taeda L.) through somatic embryogenesis has the potential to meet the increasing industrial demands for high-quality uniform raw materials. A major barrier to the commercialization of this technology is the low quality of the resulting embryos. Twenty-five newly initiated loblolly pine genotypes were followed through the process of liquid culture establishment, embryo maturation, germination, and retrieval from cryogenic storage. A maturation medium, cap...
Ref 26Cited 93
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John W.G. Cairney45
Estimated H-index: 45
,
Nanfei Xu6
Estimated H-index: 6
,
Gerald S. Pullman19
Estimated H-index: 19
... more
In production biological systems, monitoring and controlling the growth environment is possible, but assessing the metabolic competency of the organism is more difficult. Somatic embryogenesis (SE), a tissue-culture method for multiplying embryos asexually, has great potential to capture at low cost the genetic gain from breeding and genetic-engineering programs. Loblolly pine, however, has proven recalcitrant in the production of somatic embryos suitable in quality for operational use. Many sim...
Ref 20Cited 17
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Paula P. Chee1
Estimated H-index: 1
Taxusbrevifolia is the source of paclitaxel (Taxol®), an anticancer drug. A method for regeneration ofTaxus brevifolia from immature zygotic embryos via somatic embryogenesis is described. Embryogenic callus tissues were obtained by culturing immature zygotic embryos on Lloyd and McCown medium (MCM) supplemented with 160 μM 2,4-dichlorophenoxyacetic acid (2,4-D) + 5 μM benzylaminopurine (BA) + 5 μM naphthaleneacetic acid (NAA) for 4 weeks. Putative embryoids were obtained following transfer of c...
Ref 16Cited 18
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Gerald S. Pullman19
Estimated H-index: 19
(Weyerhaeuser),
Pramod K. Gupta16
Estimated H-index: 16
(Weyerhaeuser),
Roger Timmis14
Estimated H-index: 14
(Weyerhaeuser)
... more
The combination of abscisic acid (ABA) and activated carbon increased Norway spruce (Picea abies L., Karst.) cotyledonary somatic embryo yields, increased the number of genotypes forming cotyledonary embryos, caused embryos to form that exhibited improved maturation characteristics, and reduced embryo production costs. Somatic embryos increased in size, showed larger apical regions, became more zygotic-like in shape, and showed higher percentages of epicotyl development upon germination. Analyse...
Ref 37Cited 42
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G. S. Pullman1
Estimated H-index: 1
,
Y. Zhang1
Estimated H-index: 1
,
Bao H. Phan4
Estimated H-index: 4
(University of Georgia)
Somatic embryogenesis (SE), the most promising technology for the large-scale production of high-value coniferous trees from advanced breeding and genetic engineering programs, is expected to play an important role in increasing productivity, sustainability, and the uniformity of future U.S. forests. To be successful for commercial use, SE technology must work with a variety of genetically diverse trees. Initiation in loblolly pine (Pinus taeda L.), our main focus species, is often recalcitrant ...
Ref 22Cited 76
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  • Cited By (18)
2016 in Protoplasma
Neusa Steiner11
Estimated H-index: 11
(Universidade Federal de Santa Catarina),
Francine L. Farias-Soares5
Estimated H-index: 5
(Universidade Federal de Santa Catarina),
Éder C. Schmidt16
Estimated H-index: 16
(Universidade Federal de Santa Catarina)
... more
Somatic embryogenesis is a morphogenetic route useful for the study of embryonic development, as well as the large-scale propagation of endangered species, such as the Brazilian pine (Araucaria angustifolia). In the present study, we investigated the morphological and ultrastructural organization of A. angustifolia somatic embryo development by means of optical and electron microscopy. The proembryogenic stage was characterized by the proliferation of proembryogenic masses (PEMs), which are cell...
Ref 72Cited 10
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Qixiang Zhang4
Estimated H-index: 4
,
Weibo Xiang , Jin Yao ... more
Key message This paper provides an important direct proembryogenic mass initiation and plant regeneration method that might be supportive and stable for gene transformation and commercial production of Torreya grandis. Abstract Torreya grandis is an important and commer- cially valuable tree that is utilized as a source of essential oil and nuts. Propagation researches of T. grandis have been focused on micropropagation and somatic embryogenesis which derived from callus but limited data regardi...
Ref 28
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André Luis Wendt dos Santos7
Estimated H-index: 7
(University of São Paulo),
Paula Elbl4
Estimated H-index: 4
(University of São Paulo),
Bruno Viana Navarro3
Estimated H-index: 3
(University of São Paulo)
... more
GeLC–MS/MS based label free proteomic profiling was used in the large scale identification and quantification of proteins from Brazilian pine (Araucaria angustifolia) embryogenic cell (EC) lines that showed different propensities to form somatic embryos. Using a predicted protein sequence database that was derived from A. angustifolia RNA-Seq data, 2398 non-redundant proteins were identified. The log2 of the spectral count values of 858 proteins of these proteins showed a normal distribution, an...
Ref 100Cited 12
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Sharrmila Rengeswari Palanyandy3
Estimated H-index: 3
(Universiti Putra Malaysia),
Periasamy Suranthran5
Estimated H-index: 5
(Universiti Putra Malaysia),
Saikat Gantait14
Estimated H-index: 14
(Universiti Putra Malaysia)
... more
In the present study, we report the in vitro development of polyembryoids with identification of a definite stage that can be used for subsequent uniform plantlet regeneration in oil palm (Elaeis guineensis Jacq.). Induction and maturation of polyembryoids was accomplished when cell suspension culture was transferred in MS (Murashige and Skoog, Physiol Plant 15:473–497, 1962) semisolid medium consisting of 30 g L−1 sucrose and 3.5 g L−1 gelrite® devoid of any plant growth regulator. Growth and d...
Ref 23Cited 9
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2014 in New Forests
Gerald S. Pullman20
Estimated H-index: 20
(Georgia Institute of Technology),
Kylie Bucalo7
Estimated H-index: 7
(Georgia Institute of Technology)
The shift from vegetative to embryogenic growth requires tissue to enter a radically different program of development and can be studied in vitro through the development of somatic embryos. From an applied perspective somatic embryogenesis (SE) is expected to play an important role in increasing productivity, sustainability, and uniformity of future forests. For commercial use, SE technology must work with a variety of genetically diverse trees. Since the first reports of SE in Picea abies and L...
Ref 157Cited 20
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Leonardo Jo3
Estimated H-index: 3
(University of São Paulo),
André Luis Wendt dos Santos7
Estimated H-index: 7
(University of São Paulo),
Caroline A. Bueno1
Estimated H-index: 1
(University of São Paulo)
... more
Somatic embryogenesis is an important biotechnological tool in the large-scale propagation of elite genotypes and ex situ conservation of conifer species. Protocols for the induction and proliferation of embryogenic cultures (ECs) of Brazilian pine (Araucaria angustifolia (Bert.) O. Ktze) are well established, although the proper formation of mature somatic embryos (SEs) is still problematic. Thus, the identification of molecular markers for the screening of ECs able to respond to maturation con...
Ref 78Cited 24
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2016 in New Forests
Gerald S. Pullman20
Estimated H-index: 20
(Georgia Institute of Technology),
Katie Olson1
Estimated H-index: 1
(Georgia Institute of Technology),
Taylor Fischer1
Estimated H-index: 1
(Georgia Institute of Technology)
... more
Fraser fir (Abies fraseri [Pursh] Poir.) is a coniferous species native to the Southern Appalachian Mountains of the eastern United States. The species has high economic and recreational value but is vulnerable to extinction due to introduced pests and global warming. Somatic embryogenesis technology may assist in the clonal production of desired lines of Christmas trees and safekeeping of rare and valuable germplasm via cryopreservation. We have developed a highly effective medium for initiatio...
Ref 63Cited 8
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