Evaluation of Different Types of Daylily Explants for Somatic Embryogenesis

To improve somatic embryogenesis of Daylilies (Hemerocallis hybrid); six types of explants namely immature seeds, immature embryos, mature embryos, young inflorescence, ovary-sections and filaments of daylily flower buds were investigated as source of explants. Explants from field grown plants were surface sterilized and followed by culturing on MS medium supplemented with 1.5 mg/L NAA and 0.5 mg/L BAP for four weeks. Explants were scored for development of embryogenic tissue and formation of somatic embryos with/without the formation of an intermediate callus stage. Both mature and immature embryo explants produced direct development of embryogenic tissue followed by somatic embryos. Young inflorescence explants developed compact calli and produced roots around cut ends and showed no somatic embryogenesis. Ovary explants exhibited swelling and not produced embryogenesis. Production of embryogenic callus and formation of somatic embryos in filament explants depended on the sizes of flower buds. Explants from 0.5 to 1.2 mm size flower buds produced calli and formed somatic embryos while explants from sizes over 1.2 mm flower buds only non-embryogenic calli. Immature seeds failed to grow. The results indicated that immature and mature embryos and filaments from young flower buds responded better than other explants for developing somatic embryos in daylily.

Download Full-text

Polyamines during somatic embryo development in Norway spruce (Picea abies [L.])

Journal of Forest Science ◽

10.17221/84/2008-jfs ◽

2009 ◽

Vol 55 (No. 2) ◽

pp. 75-80 ◽

Cited By ~ 9

Author(s):

J. Malá ◽

M. Cvikrová ◽

P. Máchová ◽

O. Martincová

Keyword(s):

Somatic Embryogenesis ◽

Picea Abies ◽

Norway Spruce ◽

Somatic Embryos ◽

Developmental Stages ◽

Embryogenic Tissue ◽

Growth Media ◽

Mature Embryos ◽

Proliferation Medium ◽

Maturation Medium

Contents of free polyamines (putrescine, spermidine and spermine) were determined in different developmental stages of Norway spruce (<I>Picea abies</I> [L.] Karst.) somatic embryos by means of HPLC. Determinations were performed embryogenic tissue after 4 weeks of the growth on proliferation medium, after 2 and 5 weeks of the culturing on maturation medium, and 2 weeks after desiccation. Maturation of somatic embryos (after 5 weeks) was accompanied by increase of concentrations of putrescine (2.3 times) and spermidine (3.2 times). In comparison with above mentioned polyamines, spermine concentrations were significantly lower (4.3 times). Two weeks after desiccation, the concentrations of putrescine decreased 5.4 times and spermidine 2.2 times in comparison with mature embryos. To improve the efficiency of somatic embryogenesis of less responsive genotypes, the supplementation of growth media by polyamines is discussed.

Download Full-text

Commercialization potential of somatic embryogenesis in black spruce tree improvement

The Forestry Chronicle ◽

10.5558/tfc70593-5 ◽

1994 ◽

Vol 70 (5) ◽

pp. 593-598 ◽

Cited By ~ 14

Author(s):

G. W. Adams ◽

M. G. Doiron ◽

Y. S. Park ◽

J. M. Bonga ◽

P. J. Charest

Keyword(s):

Somatic Embryogenesis ◽

Clonal Propagation ◽

Black Spruce ◽

Tree Improvement ◽

Embryogenic Tissue ◽

Improvement Program ◽

Mature Embryos ◽

Somatic Plants ◽

Number Of Individuals ◽

Potential Tool

The somatic embryogenesis process was evaluated as a potential tool for operational vegetative propagation using individuals from families currently used in the J.D. Irving, Ltd. black spruce tree improvement program. Most families were responsive although the number of individuals within families capable of producing embryogenic tissue (ET) varied greatly (1–70%). Seventy-four percent of the ET clones produced mature embryos and most of these germinated. Greenhouse survival was initially low (11%) but improved in subsequent experiments to 45% as growing regimes were refined. Demonstration plantings of the resulting somatic plants were established at two sites in New Brunswick. A total of 206 clones were cryopreserved. The potential for integrating somatic embryogenesis techniques into tree improvement and stock production programs is discussed. Key words: tree improvement, somatic embryogenesis, clonal propagation, black spruce, biotechnology

Download Full-text

Can Ethylene Inhibitors Enhance the Success of Olive Somatic Embryogenesis?

Plants ◽

10.3390/plants11020168 ◽

2022 ◽

Vol 11 (2) ◽

pp. 168

Author(s):

Muhammad Ajmal Bashir ◽

Cristian Silvestri ◽

Amelia Salimonti ◽

Eddo Rugini ◽

Valerio Cristofori ◽

...

Keyword(s):

Somatic Embryogenesis ◽

Salicylic Acid ◽

Silver Nitrate ◽

Embryogenic Callus ◽

Somatic Embryos ◽

Cobalt Chloride ◽

Zygotic Embryos ◽

Ethylene Inhibitors ◽

Embryogenic Calli ◽

Stimulatory Action

An efficient in vitro morphogenesis, specifically through somatic embryogenesis, is considered to be a crucial step for the application of modern biotechnological tools for genetic improvement in olive (Olea europaea L.). The effects of different ethylene inhibitors, i.e., cobalt chloride (CoCl2), salicylic acid (SA), and silver nitrate (AgNO3), were reported in the cyclic somatic embryogenesis of olive. Embryogenic callus derived from the olive immature zygotic embryos of the cultivar Leccino, was transferred to the expression ECO medium, supplemented with the ethylene inhibitors at 20 and 40 µM concentrations. Among these, the maximum number of somatic embryos (18.6) was obtained in media containing silver nitrate (40 µM), followed by cobalt chloride (12.2 somatic embryos @ 40 µM) and salicylic acid (40 µM), which produced 8.5 somatic embryos. These compounds interfered on callus traits: white friable embryogenic calli were formed in a medium supplemented with 40 µM cobalt chloride and salicylic acid; in addition, a yellow-compact embryogenic callus appeared at 20 µM of all the tested ethylene inhibitors. The resulting stimulatory action of silver nitrate among all the tested ethylene inhibitors on somatic embryogenesis, clearly demonstrates that our approach can efficiently contribute to the improvement of the current SE protocols for olive.

Download Full-text

Pinus canariensis plant regeneration through somatic embryogenesis

Forest Systems ◽

10.5424/fs/2020291-16136 ◽

2020 ◽

Vol 29 (1) ◽

pp. eSC05

Author(s):

Ander Castander-Olarrieta ◽

Paloma Moncaleán ◽

Itziar A. Montalbán

Keyword(s):

Somatic Embryogenesis ◽

Cell Lines ◽

Somatic Embryos ◽

Embryogenic Tissue ◽

Established Cell Line ◽

Zygotic Embryos ◽

Pinus Canariensis ◽

Embryogenic Cell ◽

Embryogenic Cell Lines ◽

Established Cell

Aim of the study: To develop an efficient method to regenerate plants through somatic embryogenesis of an ecologically relevant tree species such as Pinus canariensis.Area of study: The study was conducted in the research laboratories of Neiker-Tecnalia (Arkaute, Spain).Material and methods: Green cones of Pinus canariensis from two collection dates were processed and the resulting immature zygotic embryos were cultured on three basal media. The initiated embryogenic tissues were proliferated testing two subculture frequencies, and the obtained embryogenic cell lines were subjected to maturation. Germination of the produced somatic embryos was conducted and acclimatization was carried out in a greenhouse under controlled conditions.Main results: Actively proliferating embryogenic cell lines were obtained and well-formed somatic embryos that successfully germinated were acclimatized in the greenhouse showing a proper growth.Research highlights: This is the first report on Pinus canariensis somatic embryogenesis, opening the way for a powerful biotechnological tool for both research purposes and massive vegetative propagation of this species.Keywords: acclimatization; Canary Island pine; micropropagation; embryogenic tissue; somatic embryo.Abbreviations used: embryogenic tissue (ET); established cell line (ECL); somatic embryogenesis (SE); somatic embryos (Se’s).

Download Full-text

Propagation of interior spruce by somatic embryogenesis

Canadian Journal of Forest Research ◽

10.1139/x90-234 ◽

1990 ◽

Vol 20 (11) ◽

pp. 1759-1765 ◽

Cited By ~ 57

Author(s):

F. B. Webster ◽

D. R. Roberts ◽

S. M. McInnis ◽

B. C. S. Sutton

Keyword(s):

Somatic Embryogenesis ◽

Abscisic Acid ◽

Relative Humidity ◽

Somatic Embryos ◽

Large Scale ◽

Final Height ◽

High Relative Humidity ◽

Mature Embryos ◽

Interior Spruce ◽

General Production

To apply somatic embryogenesis to clonal propagation of forest species, the technique must be applicable to a broad range of genotypes and allow efficient regeneration of phenotypically normal plants. Seventy-one lines (genotypes) of embryogenic cultures from six open-pollinated families were obtained by culturing immature embryos of interior spruce. Interior spruce represents a mixture of two closely related species, Piceaglauca (Moench) Voss and Piceaengelmannii Parry, from the interior of British Columbia where they hydridize with one another. The abscisic acid dependent developmental profile (the proportion of rooty embryos, shooty embryos, precociously germinating embryos, and mature embryos over a range of abscisic acid concentrations) differed among genotypes, but in general, production of mature somatic embryos was highest at 40 and 60 μM abscisic acid. Treatment of mature embryos with a high relative humidity treatment resulted in partial drying of the embryos and upon rehydration, markedly enhanced germination of the eight genotypes tested. Within 1 week of being placed under germination conditions, somatic embryos treated with the high relative humidity treatment showed 80–100% germination for 12 of the genotypes, and most genotypes had germination rates of greater than 40%. Survival of "emblings" (germinants from somatic embryos) following transfer to soil, acclimatization, and first season's growth in the nursery was 80% or greater for most genotypes. Over 1200 emblings were tested for nursery performance, representing the first large-scale evaluation of conifer somatic embryos under exvitro conditions. Growth rates, final height, shoot and root morphology, and frost hardiness were similar for emblings and seedlings following the first growing season. These results indicate that somatic embryogenesis can be used for the production of planting stock for a range of interior spruce genotypes.

Download Full-text

Efficient Somatic Embryogenesis and Plant Regeneration from Immature Embryos of Tapiscia sinensis Oliv., an Endemic and Endangered Species in China

HortScience ◽

10.21273/hortsci.49.12.1558 ◽

2014 ◽

Vol 49 (12) ◽

pp. 1558-1562 ◽

Cited By ~ 4

Author(s):

Yuyu Wang ◽

Faju Chen ◽

Yubing Wang ◽

Xiaoling Li ◽

Hongwei Liang

Keyword(s):

Somatic Embryogenesis ◽

Plant Regeneration ◽

Activated Charcoal ◽

Somatic Embryos ◽

Basal Medium ◽

Immature Embryos ◽

Zygotic Embryos ◽

Naphthaleneacetic Acid ◽

Embryogenic Calli ◽

Induction Rate

High-frequency somatic embryogenesis and plant regeneration were achieved from immature cotyledonary-stage embryos in the endangered plant, Tapiscia sinensis Oliv. Plant growth regulators with different concentrations and combinations on embryogenesis capacity were studied. The optimal explants for in vitro somatic embryogenesis were immature embryos in T. sinensis. A high callus induction rate of 100% was achieved on Murashige and Skoog (MS) basal medium supplemented with 1.0 mg·Ll−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5% (w/v) activated charcoal. Alternatively, a high induction rate (96.16%) of somatic embryogenesis was obtained on MS basal medium supplemented with the combination of 0.05 mg·L−1 α-naphthaleneacetic acid (NAA) and 0.2 mg·L−1 6-benzylaminopurine (6-BA), and somatic embryos proliferated fastest on the mentioned medium supplemented with 0.5% (w/v) activated charcoal and 3% (w/v) sucrose, inoculation of explants proliferating 21 times in the 23-day subculture. Of the 100 plantlets transferred to field after the acclimation, 95 (95%) survived. Based on the histocytological observations, the development of somatic embryos was similar to that of zygotic embryos. There were two accumulation peaks of starch grains in the embryogenic calli and in the globular-stage embryos, both closely related to the energy supply, and the embryoids were of multicelluar origin.

Download Full-text

Enhanced Somatic Embryo Induction of a Tree Peony, Paeonia ostii ‘Fengdan’, by a Combination of 6-benzylaminopurine (BA) and 1-naphthylacetic Acid (NAA)

Plants ◽

10.3390/plants9010003 ◽

2019 ◽

Vol 9 (1) ◽

pp. 3 ◽

Cited By ~ 2

Author(s):

Xiuxia Ren ◽

Ya Liu ◽

Byoung Ryong Jeong

Keyword(s):

Somatic Embryogenesis ◽

Somatic Embryos ◽

Zygotic Embryo ◽

Zygotic Embryos ◽

Ms Medium ◽

Hypocotyl Explants ◽

Cotyledon Explants ◽

Embryo Induction ◽

Naphthylacetic Acid ◽

Better Than

Somatic embryogenesis is a preferred method for vegetative propagation due to its high propagation efficiency. In this study, zygotic embryos, cotyledons, and hypocotyls of Paeonia ostii ‘Fengdan’ were used as the explant to induce somatic embryogenesis. The results showed that a combination of 0.5 mg·L−1 thidiazuron (TDZ) and 0.5 mg·L−1 2,4-dichlorophenoxyacetic acid (2,4-D) was effective in inducing somatic embryos from the zygotic embryo and cotyledon explants. Hypocotyls only formed somatic embryos on Murashige and Skoog (MS) medium supplemented with both 0.5 mg·L−1 TDZ and 0.5 mg·L−1 1-naphthylacetic acid (NAA). Moreover, the compact callus was effectively produced from zygotic embryo, cotyledon, and hypocotyl explants in medium supplemented with a combination of 3.0 mg·L−1 6-benzylaminopurine (BA) and 1.0 mg·L−1 NAA, and then converted into somatic embryos in the same medium, and the ratio of the explants with embryo induction and number of embryos induced per explant were much higher than those induced by 0.5 mg·L−1 TDZ and either 0.5 mg·L−1 2,4-D or 0.5 mg·L−1 NAA. The MS medium was better than the woody plant medium (WPM) for inducing somatic embryos from zygotic embryo and hypocotyl explants, whereas the WPM was better than the MS medium for somatic embryogenesis induction from cotyledon explants. All of the somatic embryos developed well into mature embryos on their respective media supplemented with both 3.0 mg·L−1 BA and 1.0 mg·L−1 NAA. Overall, the protocols for indirect somatic embryogenesis from zygotic embryo, cotyledon, and hypocotyl of P. ostii ‘Fengdan’ were successfully established, which can greatly facilitate their propagation and breeding processes.

Download Full-text

Changes in biochemistry and histochemical characteristics during somatic embryogenesis in Ormosia henryi Prain

10.1101/2020.09.21.307009 ◽

2020 ◽

Author(s):

Gaoyin Wu ◽

Xiaoli Wei ◽

Xiao Wang ◽

Xian Liang ◽

Yi Wei

Keyword(s):

Reactive Oxygen Species ◽

Somatic Embryogenesis ◽

Enzyme Activities ◽

Soluble Protein ◽

Somatic Embryos ◽

Soluble Sugar ◽

Endogenous Hormones ◽

Oxygen Species ◽

Mature Embryos ◽

Protein Granules

ABSTRACTMature embryos were used as an explant for embryogenic callus (EC) induction, and then EC was further developed to form somatic embryos during somatic embryogenesis (SE) of Ormosia henryi Prain; however, some mature embryos could induced non-embryogenic callus (NEC), browning callus (BC) or snowflake callus (SC). These phenomena might be related to the biochemical and histochemical differences during somatic embryo induction. The present study was conducted to analyze the biochemical events and histochemical changes at different SE stages in 0. henryi. The contents of soluble sugar, starch, soluble protein, H2O2, and endogenous hormones and the activities of polyphenoloxidase (PPO), superoxide dismutase (SOD), ascorbate peroxidase (APX), peroxidase (POD) and catalase (CAT) were measured at different SE stages, such as EC, globular embryo (GE), and cotyledon embryo (CE), and in abnormal tissue, such as NEC, BC, and SC. The results showed that the contents of soluble sugar and starch; the activities of PPO, SOD, APX and POD; and the ratios of indole-3-acetic acid/abscisic acid (IAA/ABA), IAA/gibberellins (IAA/GAs), auxin /GAs (AUX/GAs), and AUX/ABA decreased gradually at different SE stages. In contrast, the contents of soluble protein, H2O2, all endogenous hormones gradually increased. However, CAT activity and the ratios of IAA/cytokinins (IAA/CKs), AUX/CKs, ABA/CKs, and GAs/CKs first increased and then decreased. The high contents of GAs and ABA, high ratios of ABA/CKs and GAs/CKs and low ratios of IAA/ABA, IAA/GAs, AUX/GAs and AUX/ABA were responsible for the inability of the callus to form EC. The low enzyme activities, low contents of energy substances and H2O2 were related to NEC formation. The high contents of soluble sugar, H2O2, AUX, CKs and PPO activity and the low content of soluble protein were the basic causes of BC formation. The high-energy substances contents and low activities of SOD and POD facilitated SC formation. Histochemical observation showed that starch granule staining gradually lightened with SE development, but protein granules were darkly stained. Compared with EC, starch and protein granules were stained darker in SC, and lighter in NEC and BC. These results showed that energy substances were the material basis of SE, which affected enzyme activities, regulated reactive oxygen species (ROS) metabolism, and thus regulated the morphogenesis and development of somatic embryos. In addition, the contents and ratios of endogenous hormones affected the dedifferentiation, dedifferentiation and embryogenesis of somatic cells. To induce EC from mature embryos and further develop their formation into somatic embryos, it is necessary to adjust the energy supply and hormone ratio in the medium.One-sentence summarySomatic embryogenesis and abnormal callus tissues formation of Ormosia henryi Prain were associated with energy substances, reactive oxygen species, enzyme activities and endogenous hormones, as well as histochemical characteristics.

Download Full-text

Regeneration of Bigleaf Magnolia by Somatic Embryogenesis

HortScience ◽

10.21273/hortsci.28.6.672 ◽

1993 ◽

Vol 28 (6) ◽

pp. 672-673 ◽

Cited By ~ 8

Author(s):

S.A. Merkle ◽

B.A. Watson-Pauley

Keyword(s):

Acetic Acid ◽

Somatic Embryogenesis ◽

Somatic Embryos ◽

Casein Hydrolysate ◽

Basal Medium ◽

Induction Medium ◽

Immature Seeds ◽

Dichlorophenoxy Acetic Acid ◽

Adventive Embryos

Bigleaf magnolia (Magnolia macrophylla Michx.) cultures were initiated from immature seeds on an induction medium containing 9.0 μm 2,4-D, 1.1μm BA, and 1 g casein hydrolysate/liter. After 2 months on induction medium, one culture produced adventive embryos. Clumps of embryos transferred to liquid induction medium proliferated as nodules, which grew in diameter, but failed to produce embryos while maintained in induction medium. Nodules transferred to basal medium produced clumps of somatic embryos, which continued to produce repetitive embryos with monthly transfer to fresh basal medium. Individual embryos transferred to basal medium lacking casein hydrolysate germinated and leaves expanded. Plantlets derived from these embryos were transferred to potting mix and acclimatized to greenhouse conditions. Chemical names used: (2,4-dichlorophenoxy)acetic acid (2,4-D); N -(phenylmethyl)-lH-purin-6-amine (BA).

Download Full-text

Identification of Serk Gene from Bract Derived Embryogenic and Non - Embryogenic Calli of Four Diploid Banana Cultivars from South India

Journal of Agricultural Studies ◽

10.5296/jas.v5i4.12213 ◽

2017 ◽

Vol 5 (4) ◽

pp. 161

Author(s):

Smitha P D ◽

Binoy K R ◽

Ashalatha S Nair

Keyword(s):

Somatic Embryogenesis ◽

Somatic Embryos ◽

Cell Formation ◽

Molecular Size ◽

Receptor Kinase ◽

Embryogenic Calli ◽

Embryogenic Cell ◽

Diploid Banana ◽

Somatic Embryogenesis Receptor Kinase ◽

Similar Intensity

Somatic embryogenesis receptor kinase (SERK) gene is known to be a marker of somatic embryogenesis in several plant species. The present study reported the presence of SERK gene from bract derived embryogenic calli bearing somatic embryos. The analysis of the expression pattern of the SERK gene during embryogenic cell formation and somatic embryogenesis revealed that SERK expression continued during pro embryogenic mass formation. In the present study the amplified product of cDNA from the somatic embryos has molecular size 1459 bp. The non- embryogenic callus also showed the presence of faint bands. In all the samples the amplified product from β - actin primer showed bands of 650 bp with similar intensity in both the embryogenic and non- embryogenic samples.

Download Full-text