scholarly journals THE EFFECT OF PICLORAM AND LIGHT ON SOMATIC EMBRYOGENESIS REGENERATION OF PINEAPPLE

2012 ◽  
Vol 13 (2) ◽  
pp. 43 ◽  
Author(s):  
Ika Roostika ◽  
Nurul Khumaida ◽  
Ika Mariska ◽  
Gustaaf Adolf Wattimena
Keyword(s):  
Somatic Embryogenesis ◽  
Butyric Acid ◽  
Somatic Embryos ◽  
Callus Formation ◽  
Light Condition ◽  
Cell Polarization ◽  
Mass Propagation ◽  
Benzyl Adenine ◽  
Indole Butyric Acid ◽  
Embryogenic Tissues

<p>Smooth Cayenne is the largest pineapple type cultivated in Indonesia, but its vegetative planting materials for mass propagation are limited. Somatic embryogenesis is a potential method to be applied. The aim of this study was to investigate the somatic embryogenesis regeneration under the effect of picloram and light. Callus formation was induced by picloram (21, 41 and 62 μM) added with 9 μM thidiazuron. The calli were transferred onto MS or Bac medium  enriched with N-organic compounds with or without addition of 21 μM picloram under dark or light condition. The compact calli were subcultured onto MS medium supplemented with 4.65 μM kinetin, while the friable calli were  transferred onto BIG medium (modified MS + 1.1 μM benzyl adenine + 0.9 μM indole butyric acid + 0.09 μM giberelic acid) or B medium (MS + 0.018 mM benzyl adenine). The results showed that the events of somatic embryogenesis were started from cell polarization, asymmetrical division, proembryo formation as  embryogenic tissues and friable embryogenic tissues, and embryo development. The best treatment for callus induction was 21 μM picloram. The addition of 21 μM picloram on N-organic enriched medium and the use of light condition  proliferated embryogenic calli. The N-organic enriched Bac medium and light condition yielded the highest number of mature somatic embryos (17 embryos per<br />explant in 2 months). The B medium was better than BIG medium to develop  somatic embryos from friable embryogenic tissues. The somatic embryogenesis method presented is potential for pineapple mass propagation and artificial seed<br />production.</p><p>Abstrak Bahasa Indonesia</p><p>Smooth Cayenne merupakan kultivar nenas yang banyak dibudidayakan di  Indonesia, namun ketersediaan benih untuk perbanyakan massal masih terbatas. Embriogenesis somatikadalah metode yang potensial untuk produksi bibit secara massal. Tujuan penelitian adalah untuk mempelajari pengaruh pikloram dan pencahayaan terhadap regenerasi embriogenesis somatik nenas. Kalus diinduksi menggunakan pikloram (21, 41, dan 62 μM) dan penambahan thidiazuron 9 μM. Selanjutnya, kalus dipindahkan ke media MS atau Bac yang diperkaya dengan<br />senyawa N-organik dengan atau tanpa penambahan pikloram 21 μM dalam kondisi gelap atau dengan pencahayaan. Kalus kompak disubkultur pada media MS yang mengandung kinetin 4,65 μM, sedangkan kalus remah dipindahkan ke media BIG (MS modifikasi + bensil adenin 1.1 μM + indole butyric acid 0,9 μM + giberelic acid 0,09 μM) atau media B (MS + bensil adenin 0,018 μM). Hasil penelitian  menunjukkan bahwa tahapan embriogenesis somatik diawali dengan polarisasi sel, pembelahan asimetris, pembentukan proembrio sebagai jaringan embriogenik dan<br />jaringan embriogenik remah, serta perkembangan embrio. Perlakuan terbaik untuk induksi kalus adalah pikloram 21 μM. Penambahan pikloram 21 μM pada media yang diperkaya dengan senyawa N-organik mampu meningkatkan jumlah kalus<br />embriogenik. Media Bac yang diperkaya senyawa N-organik dan kondisi pencahayaan menghasilkan jumlah embrio somatik dewasa terbanyak (17 embrio per eksplan dalam 2 bulan). Media B lebih baik daripada media BIG untuk regenerasi embrio somatik dari jaringan embriogenik remah. Metode embriogenesis somatik yang dihasilkan dari penelitian ini berpotensi diterapkan untuk<br />perbanyakan massal dan produksi benih nenas.</p>

scholarly journals THE EFFECT OF PICLORAM AND LIGHT ON SOMATIC EMBRYOGENESIS REGENERATION OF PINEAPPLE

2012 ◽  
Vol 13 (2) ◽  
pp. 43
Author(s):  
Ika Roostika ◽  
Nurul Khumaida ◽  
Ika Mariska ◽  
Gustaaf Adolf Wattimena
Keyword(s):  
Somatic Embryogenesis ◽  
Butyric Acid ◽  
Somatic Embryos ◽  
Callus Formation ◽  
Light Condition ◽  
Cell Polarization ◽  
Mass Propagation ◽  
Benzyl Adenine ◽  
Indole Butyric Acid ◽  
Embryogenic Tissues

<p>Smooth Cayenne is the largest pineapple type cultivated in Indonesia, but its vegetative planting materials for mass propagation are limited. Somatic embryogenesis is a potential method to be applied. The aim of this study was to investigate the somatic embryogenesis regeneration under the effect of picloram and light. Callus formation was induced by picloram (21, 41 and 62 μM) added with 9 μM thidiazuron. The calli were transferred onto MS or Bac medium  enriched with N-organic compounds with or without addition of 21 μM picloram under dark or light condition. The compact calli were subcultured onto MS medium supplemented with 4.65 μM kinetin, while the friable calli were  transferred onto BIG medium (modified MS + 1.1 μM benzyl adenine + 0.9 μM indole butyric acid + 0.09 μM giberelic acid) or B medium (MS + 0.018 mM benzyl adenine). The results showed that the events of somatic embryogenesis were started from cell polarization, asymmetrical division, proembryo formation as  embryogenic tissues and friable embryogenic tissues, and embryo development. The best treatment for callus induction was 21 μM picloram. The addition of 21 μM picloram on N-organic enriched medium and the use of light condition  proliferated embryogenic calli. The N-organic enriched Bac medium and light condition yielded the highest number of mature somatic embryos (17 embryos per<br />explant in 2 months). The B medium was better than BIG medium to develop  somatic embryos from friable embryogenic tissues. The somatic embryogenesis method presented is potential for pineapple mass propagation and artificial seed<br />production.</p><p>Abstrak Bahasa Indonesia</p><p>Smooth Cayenne merupakan kultivar nenas yang banyak dibudidayakan di  Indonesia, namun ketersediaan benih untuk perbanyakan massal masih terbatas. Embriogenesis somatikadalah metode yang potensial untuk produksi bibit secara massal. Tujuan penelitian adalah untuk mempelajari pengaruh pikloram dan pencahayaan terhadap regenerasi embriogenesis somatik nenas. Kalus diinduksi menggunakan pikloram (21, 41, dan 62 μM) dan penambahan thidiazuron 9 μM. Selanjutnya, kalus dipindahkan ke media MS atau Bac yang diperkaya dengan<br />senyawa N-organik dengan atau tanpa penambahan pikloram 21 μM dalam kondisi gelap atau dengan pencahayaan. Kalus kompak disubkultur pada media MS yang mengandung kinetin 4,65 μM, sedangkan kalus remah dipindahkan ke media BIG (MS modifikasi + bensil adenin 1.1 μM + indole butyric acid 0,9 μM + giberelic acid 0,09 μM) atau media B (MS + bensil adenin 0,018 μM). Hasil penelitian  menunjukkan bahwa tahapan embriogenesis somatik diawali dengan polarisasi sel, pembelahan asimetris, pembentukan proembrio sebagai jaringan embriogenik dan<br />jaringan embriogenik remah, serta perkembangan embrio. Perlakuan terbaik untuk induksi kalus adalah pikloram 21 μM. Penambahan pikloram 21 μM pada media yang diperkaya dengan senyawa N-organik mampu meningkatkan jumlah kalus<br />embriogenik. Media Bac yang diperkaya senyawa N-organik dan kondisi pencahayaan menghasilkan jumlah embrio somatik dewasa terbanyak (17 embrio per eksplan dalam 2 bulan). Media B lebih baik daripada media BIG untuk regenerasi embrio somatik dari jaringan embriogenik remah. Metode embriogenesis somatik yang dihasilkan dari penelitian ini berpotensi diterapkan untuk<br />perbanyakan massal dan produksi benih nenas.</p>

scholarly journals EFFECT OF DIFFERENT SOURCES OF PLANT GROWTH REGULATOR ON THE INDUCTION AND DEVELOPMENT OF MANGOSTEEN SOMATIC EMBRYOS

2017 ◽  
Vol 17 (1) ◽  
pp. 9
Author(s):  
Yosi Zendra Joni ◽  
Riry Prihatini ◽  
Darda Efendi ◽  
Ika Roostika
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Growth ◽  
Embryogenic Callus ◽  
Plant Growth Regulator ◽  
Somatic Embryos ◽  
Casein Hydrolysate ◽  
Malt Extract ◽  
Mass Propagation ◽  
Embryogenic Callus Induction

<p>Somatic embryogenesis is a technique for regenerating embryos derived from somatic cells of various plant species. This technique along with the utilization of plant growth regulator (PGR) might benefit for mass propagation and improvement of plant species through biotechnological tools. The study aimed to determine the effect of different plant growth regu-lators, namely 6-benzyladenine (BA) and thidiazuron (TDZ) on the embryogenic callus induction as well as casein hydrolysate and malt extract on the somatic embryo development of mangosteen. The explants used were in vitro young stems of mangosteen clone Leuwiliang. This study consisted of two experiments, namely induction of embryogenic callus and formation of somatic embryo. The first experiment was arranged as factorial in a completely randomized design with BA (0 and 0.7 mg l-1) as the first factor and TDZ (0, 0.1, 0.5 and 1.0 mg l-1) as the second factor. The second experiment consisted of four treatments, i.e. casein hydrolysate and malt extract at the rate of 500 and 1,000 mg l-1. The results showed that the best medium for embryogenic callus induction was MS supplemented with 0.1 mg l-1 TDZ, which resulted semifriable calli. Casein hydrolysate and malt extract could not induce the formation of somatic embryos. After two times subcultures on the same MS medium supplemented with 0.5 mg l-1 TDZ and 0.7 mg l-1 BA, a total of 33.8 somatic embryos per explant was induced. The successful somatic embryogenesis would support mangosteen breeding and in vitro mass propagation program.</p>

scholarly journals In vitro ROOTING OF TENERA HYBRID OIL PALM (Elaeis guineensis Jacq.) PLANTS1

2018 ◽  
Vol 41 (4) ◽  
Author(s):  
Marlúcia Souza Pádua ◽  
Raíssa Silveira Santos ◽  
Luciano Vilela Paiva ◽  
Vanessa Cristina Stein ◽  
Luciano Coutinho Silva
Keyword(s):  
Somatic Embryogenesis ◽  
Growth Regulators ◽  
Oil Palm ◽  
Somatic Embryos ◽  
Large Scale ◽  
Elaeis Guineensis ◽  
Germination Percentage ◽  
Shoot Length ◽  
Indole Butyric Acid

ABSTRACT Oil palm is a woody monocot of economic importance due to high oil production from its fruits. Currently, the conventional method most used to propagate oil palm is seed germination, but success is limited by long time requirements and low germination percentage. An alternative for large-scale propagation of oil palm is the biotechnological technique of somatic embryogenesis. The rooting of plants germinated from somatic embryos is a difficult step, yet it is of great importance for later acclimatization and success in propagation. The aim of this study was to evaluate the effect of the auxins indole acetic acid (IAA) and indole butyric acid (IBA) on the rooting of somatic embryos of Tenera hybrid oil palm. Plants obtained by somatic embryogenesis were inoculated in modified MS medium with 10% sucrose and 0.6% agar and supplemented with IAA or IBA at concentrations of 5 µM, 10 µM, and 15 µM, and the absence of growth regulators. After 120 days, the presence of roots, root type, length of the longest root, number of roots, number of leaves, and shoot length were analyzed. Growth regulators were favorable to rooting; plants cultivated with IBA growth regulator at 15 µM showed higher rooting percentage (87%) and better results for the parameters of number of roots (1.33) and shoot length (9.83).

scholarly journals Bioproduction of Diosgenin in Callus Cultures of Balanites aegyptiaca: Effect of Growth Regulators, Explants and Somatic Embryogenesis

2006 ◽  
Vol 1 (3) ◽  
pp. 1934578X0600100
Author(s):  
Bishnu P. Chapagain ◽  
Vinod Saharan ◽  
Dan Pelah ◽  
Ram C. Yadav ◽  
Zeev Wiesman
Keyword(s):  
Somatic Embryogenesis ◽  
Growth Regulators ◽  
Embryogenic Callus ◽  
Somatic Embryos ◽  
Callus Formation ◽  
Basal Medium ◽  
Callus Cultures ◽  
Suspension Cultures ◽  
Balanites Aegyptiaca ◽  
Basal Media

This study describes the effects of plant growth regulators, explants, and somatic embryogenesis on in vitro production of the steroidal sapogenin, diosgenin, in callus cultures of the Balanites aegyptiaca (L.) Del.(desert date). Root, shoot, hypocotyl, and epicotyl callus culture of B. aegyptiaca, were raised on MS basal media supplemented with various combinations of either 2,4-D and NAA alone, or with BAP. The diosgenin content (on a dry weight basis) was found to be highest when calli were cultured in MS basal medium supplemented with 1.0 mg l−1 2,4-D alone and/or in combination with 0.5 mg l−1 BAP. However, the callus growth was highest in media supplemented with 2.5 or 3.0 mg l−1 2,4-D. MS basal media supplemented with 2,4-D 2.5 mg l−1 alone and in combination with 0.5 mg l−1 BAP induced pre-embryogenic callus formation on root cultures. When these pre-embryogenic callus cultures were used to establish cell suspension cultures, two growth densities were obtained in embryogenic suspension cultures, inducing clusters of somatic embryos at various stages of development. The maximum number of somatic embryos were obtained at the fifth week on the medium supplemented with 1.0 mg l−1 2,4-D. However, the diosgenin content in these somatic cells was found to be lower compared to the explant calluses. This study revealed that production of diosgenin in callus cultures of B. aegyptiaca is possible, but the amount is significantly affected by the growth regulators, type of explants, and somatic embryogenesis.

Histological studies of cellular differentiation during somatic embryogenesis of coconut plumule-derived calli

2015 ◽  
Vol 43 (3) ◽  
Author(s):  
K. Lakshmi Jayaraj ◽  
U. Bhavyashree ◽  
T.P. Fayas ◽  
K.K. Sajini ◽  
M.K. Rajesh ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Callus Formation ◽  
Histological Study ◽  
Shoot Meristem ◽  
Vascular Bundles ◽  
Histological Studies ◽  
Meristematic Cells ◽  
Meristem Development

<div><table cellspacing="0" cellpadding="0" align="center"><tbody><tr><td align="left" valign="top"><p>Since coconut is   one of the most recalcitrant species to generate <em>in vitro</em>, it is   necessary to study in detail about the cellular changes that occur during   somatic embryogenesis to enhance our knowledge about this phenomenon. In the   present study, coconut plumular tissues, the shoot meristem including leaf   primordia, were used as explants for <em>in vitro </em>regeneration studies.   Histological studies were carried out in different stages of plumule culture.   No noticeable growth was observed in 15 days old cultures. After 30 days,   meristematic cells could be identified. Abundance of meristematic cells,   foremost to the development of callus structures, was observed after 45 days.   After 75 days, globular friable calli were formed and histological studies   revealed the presence of meristematic centers which eventually formed somatic   embryos. The histological study of matured somatic embryos formed after 120   days of callus initiation showed a clear meristematic zone of parenchyma   cells, surrounded by vascular bundles. Histological studies, carried out for   certain abnormalities like compact calli, abnormal somatic embryoids with   rudimentary shoots and multiplied roots, revealed the presence of intact   cotyledonary leaves which seemed to inhibit the apical meristem development   of somatic embryoids. The presence of vascular bundles in the early stages of   callus formation might lead to the direct formation of meristemoids. These   results could aid future studies leading to enhanced control of the somatic   embryogenic process and greater efficiency of somatic embryo and plantlet   formation in coconut.</p></td></tr></tbody></table></div>

scholarly journals The evaluation of java fine flavor cocoa propagation through somatic embryogenesis technique for germplasm preservation

2021 ◽  
Vol 306 ◽  
pp. 01056
Author(s):  
Sulistyani Pancaningtyas
Keyword(s):  
Somatic Embryogenesis ◽  
Embryogenic Callus ◽  
Somatic Embryos ◽  
Callus Formation ◽  
Basic Structure ◽  
Embryonic Cells ◽  
Regeneration Rate ◽  
Germplasm Preservation ◽  
Embryogenic Callus Formation ◽  
Secondary Somatic Embryos

Somatic embryogenesis is one of the newest technology that applied for the mass production of cocoa. This research aims to evaluate the regeneration rate of somatic embryos through somatic embryogenesis propagation techniques on java fine flavor cocoa. Cultivars in this study are ICCRI 01, ICCRI 02, DR 1, DR 2, DRC 16, DR 38, PNT 16, and PNT 30. Observations include parameters to determine the percentage of primary callus and embryogenic callus formation and the number of somatic embryos produced. Based on data, the ability of callus to produce primary embryos is highly dependent on plant cultivars and explant sources. Five cultivars showed a higher regeneration rate using explants from the petal part, while the rest showed a higher regeneration rate using explants from the staminode section. Embryogenic callus from each cacao cultivar has the same basic structure: a nodular friable structure consisting of many embryonic cells. Some fine flavor cacao cultivars that were able to produce callus and primary somatic embryos could not produce secondary somatic embryos and plantlets. However, two cultivars, which had low potential in producing primary embryos, had the high ability to produce secondary somatic embryos and develop into plantlets.

scholarly journals INDUCTION AND MORPHOLOGICAL CHARACTERISTICS ASSOCIATED WITH EMBRYOGENESIS OF OENANTHE STOLONIFERA DC.

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 693e-693
Author(s):  
Ji-Weon Lee ◽  
Byoung-Yil Lee
Keyword(s):  
Somatic Embryogenesis ◽  
Embryogenic Callus ◽  
Somatic Embryos ◽  
Callus Formation ◽  
Morphological Characteristics ◽  
Liquid Media ◽  
Friable Embryogenic Callus ◽  
Development And Differentiation ◽  
Rapid Mass ◽  
Embryogenic Callus Formation

The study was carried out to examine the appropriate media, explant sources, and suitable growth regulators for somatic embryogenesis to establish a rapid mass production system via somatic embryogenesis in Oenanthe stolonifera DC. Modified MS media containing higher concentrations of NO3-N were more effective for the formation and development of the somatic embryos from embryogenic callus. Liquid media were more effective for the production of somatic embryos than solidified media. Immature florets were found to be the most competent explant sources for embryogenic callus formation. 2,4-D at 1mg/l was highly effective for the formation of embryogenic callus but inhibitory for the development and differentiation of somatic embryo. Somatic embryos were developed from the translucent and friable embryogenic callus. Addition of BA promoted the callus growth synefgistically with NAA and 2,4-D, but the production of embryogenic callus was inhibited by BA.

scholarly journals Plant Regeneration through Somatic Embryogenesis from Leaf Sheath Derived Callus of Sugarcane (Saccharum officinarum L.) var. Isd -16

2011 ◽  
Vol 21 (2) ◽  
pp. 143-149 ◽  
Author(s):  
Mohashweta Roy ◽  
M. Hossain ◽  
A. Biswas ◽  
M. K. Biswas ◽  
R. Islam
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryos ◽  
Callus Formation ◽  
Saccharum Officinarum ◽  
Leaf Sheath ◽  
Coconut Water ◽  
Friable Callus ◽  
Light Yellow ◽  
Half Strength

Leaf sheath explants of an indigenous variety Isd-16 of sugarcane (Saccharum officinarum L.) produced light yellow friable callus after culturing on to MS with 2,4-D (2 - 4 mg/l) and NAA (3 - 5 mg/l) singly. Callus formation was the maximum on MS + 3 mg/l 2,4-D. Callus underwent embryogenesis producing huge number of somatic embryos when subcultured on MS with 15 - 30 mg/l             L-proline, 3 mg/l 2,4-D + 5 - 10% coconut water (v/v) and 3 mg/l 2,4-D + 10% CW  (v/v) + 300 - 500 mg/l CH. L-proline significantly enhanced somatic embryo-genesis and 25 mg/l L-proline in MS was the best culture medium formulation. Most of the somatic embryos germinated and developed plantlets after 1 - 2 weeks of incubation in proline-supplimented medium. On the other hand, maturation and germination of embryos were achieved on half-strength MS with or without 0.25 - 1.0 mg/l L-proline, and 5% coconut water (v/v). Somatic embryos derived plantlets were then successfully transferred to natural condition through successive phages of acclimation.   Key words: Plant regeneration, Leaf sheath, Somatic embryogenesis, Sugarcane   D. O. I. 10.3329/ptcb.v21i2.10237   Plant Tissue Cult. & Biotech. 21(2): 143-149, 2011 (December)

scholarly journals Plant regeneration through somatic embryogenesis from calli derived from leaf bases of Laurus nobilis L. (Lauraceae)

2015 ◽  
Vol 24 (2) ◽  
pp. 213-221 ◽  
Author(s):  
Ahmad H Al Gabbiesh ◽  
M Ghabeish ◽  
I H ◽  
M Kleinwächter ◽  
D Selmar
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryos ◽  
Root Formation ◽  
Callus Formation ◽  
Germplasm Conservation ◽  
Laurus Nobilis ◽  
The Media ◽  
Regenerated Plantlets ◽  
Culture Technology

Somatic embryogenesis was induced in embryo culture on half MS medium supplemented with NAA (8 mg/l) as the sole plant growth regulator after incubation of the media in the refrigerator at 4°C for two weeks to promote callus induction and somatic embryogenesis in Laurus nobilis. Both embryogenetic calli and somatic embryos were induced in the above selected medium. Embryo growth and development were stimulated by separation of embryos successfully from embryo clusters and transferred onto fresh half MS. Among the selected explants, only leaf bases were found to respond actively to plant regeneration, especially in inducing callus formation and in sustaining faster callus growth. Root formation of regenerated plantlets tended to decrease with time on regeneration media. Overall, 75% of the plantlets derived from the callus survived in the greenhouse; and they all grew to phenotypically normal plants. This procedure will enable the use of regeneration tissue culture technology for germplasm conservation of L. nobilis, a plant of high medicinal and commercial value.Plant Tissue Cult. & Biotech. 24(2): 213-221, 2014 (December)

scholarly journals 119 Rapid Somatic Embryogenesis and Plant Regeneration in American Ginseng (Panax quiquefolium L.)

HortScience ◽  
1999 ◽  
Vol 34 (3) ◽  
pp. 462B-462
Author(s):  
X. Wang ◽  
J.T.A. Proctor ◽  
S. Krishna Raj ◽  
P.K. Saxena
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
American Ginseng ◽  
Limiting Factor ◽  
Efficient Production ◽  
Cool Temperature ◽  
Mass Propagation ◽  
Regeneration System ◽  
Efficient System ◽  
Pharmacologically Active

Ginseng is a very valuable agricultural species grown for its root, which contains pharmacologically active constituents. One limiting factor for expansion of ginseng production is an efficient method for mass propagation. Currently, seeding is the principal method of propagating ginseng, but the embryo of ginseng seeds at harvest is immature. A stratification schedule consisting of a cool-warm-cool temperature treatment over 18-22 months is required for embryo development and seed germination. An alternative for the efficient production of ginseng is mass propagation through the use of in vitro culture techniques. The objective of this work was to develop a highly efficient system for regeneration of ginseng. The efficacy of three auxins, viz. 2,4-D, NAA and dicamba, were compared for the induction of somatic embryogenesis in American ginseng. Somatic embryos formed on ginseng cotyledonary, zygotic embryo, and shoot explants after 8 weeks of induction by the auxins. Significantly more somatic embryos were induced by culture of any of the ginseng explants on media supplemented with 5 μmol·L-1 2,4-D than any other auxin treatment. Histological and SEM studies confirmed that the regenerants were somatic embryos. Somatic embryos germinated and developed into normal plants in 3-6 months. The development of a regeneration system for ginseng using somatic embryogenesis is a necessary first step for mass propagation and the improvement of American ginseng.

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