共查询到20条相似文献,搜索用时 31 毫秒
1.
Summary Explants derived from adventitious buds, rhizomes, stems, and leaves of a medicinal plant, Polygonatum cyrtonema, were studied for plantlet regeneration, and only adventitious bud explants were able to be regenerated into plantlets. Regeneration
was also accompanied by the formation of rhizome-like tissue, the medicinal portion of the plant. The optimum hormone combination
for plantlet regenertion was 4.44 μM benzyladenine plus 2.26 μM 2,4-dichlorophenoxyacetic acid, at which new adventitious buds were obtained from 96.6% of the adventitious bud explants,
with an average of 5.2 buds per explant. The best medium for root induction was half-strength Murashige and Skoog medium with
4.57 μM α-naphthaleneacetic acid, as 92% of regenerated buds rooted. Regenerated plantlets were successfully transferred to a greenhouse
with 86% survival. Histological observation indicated that new adventitious buds originated from the superficial meristematic
cell cluster of the granular callus induced from adventitious bud explants via organogenesis. 相似文献
2.
Kimberly A. Pickens Jan Wolf Jameś M. Affolter Hazel Y. Wetzstein 《In vitro cellular & developmental biology. Plant》2006,42(4):348-353
Summary The bromeliad Tillandsia eizii is a stricking species with large, colorful, and persistent inflorescences that can reach 1 m in length. The value of this
plant as an ornamental and its importance in cultural and religious activities has led to its overcollection in the wild.
Clonal propagation via tissue culture may be a means to repopulate native stands while meeting the demands for this species
as an ornamental and ceremonial plant. Adventitious bud proliferation was induced from axenically germinated scedling material.
Parameters evaluated were the age of explant material at the time of transfer onto bud-induction medium, the concentration
of plant growth regulators, and the period of exposure to induction medium. Light and seanning electron microscopy (SEM) established
the origin and development of buds. Twelve-week-old seedling explants rapidly initiated adventitious buds after a 30-d induction
period on shoot-initiation medium. Adventitious buds were induced in 40% of the explants placed on media with 2 mg l−1 6-benzylaminopurine (BA) (8.88 μM) plus 0.1 mg l−1 α-naphthaleneacetic acid (NAA) (0.54 μM) with some cultures becoming highly prolific after repeated subeulture. Shoots elongated in proliferating cultures, and plants
were successfully acclimatized and planted into the greenhouse. The results indicate that tissue culture may be used as a
means to propagate this epiphytic bromeliad species, which is being seriously affected by deforestation and habitat destruction.
In addition, adventitious bud proliferation can provide a means to propagate superior genotypes. 相似文献
3.
Hiroshi Kamada Yoshinobu Tachikawa Tsutomu Saitou Hiroshi Harada 《Plant cell reports》1995,14(10):611-615
Summary To clarify that the presence of Ri T-DNA genes are not prerequisite for the light-induced bud formation in horseradish (Armoracia rusticana) hairy roots, leaf and root segments of nontransformed horseradish plants were used as explants. Bud formation from nontransformed tissues was observed in hormone-free medium under 16 h daylight conditions, but not under continuous darkness. To investigate the effects of growth regulators on bud formation, leaf and root explants were treated with auxin (1-naphthaleneacetic acid; NAA) and / or cytokinin (6-benzyl-aminopurine; BA). The most effective treatment in the dark to stimulate bud formation was BA at 1 mg·1-1. These results show that adventitious bud formation in horseradish can be induced by light and growth regulators, and especially cytokinin, may be involved in bud formation, irrespective of whether the tissues were transformed with Ri T-DNA.Abbreviations BA
6-benzyl-aminopurine
- NAA
1-Naphthaleneacetic acid
- MS
Murashige & Skoog (1962) medium 相似文献
4.
Summary Callus induction, adventitious shoot and root formation, and somatic embryogenesis were investigated in root, cotyledon and mesocotyl cultures ofBellevalia romana (L.) Rchb. grown on a synthetic nutrient medium containing different plant hormones. The combination of naphtaleneacetic acid plus benzylaminopurine was very effective in causing callus growth and plant regeneration from mesocotyl explants. On the contrary 2,4-dichlorophenoxyacetic acid caused suppression of shoot bud development in the same type of callus. Both cotyledon and root derived calli showed a low growth rate and did not regenerate shoots but only roots. Differentiation of somatic embryos which eventually developed into plantlets was promoted by 2,4-dichlorophenoxyacetic acid in suspension cultures. The results are discussed in relation to studies on nuclear behaviour during different morphogenetic pathways. 相似文献
5.
María Laura Vidoz Pablo Klusacek Hebe Yolanda Rey Luis Amado Mroginski 《Plant Cell, Tissue and Organ Culture》2006,86(1):111-115
In vitro protocols for plant regeneration of Arachis correntina through both somatic embryogenesis and organogenesis were developed using immature leaves as explants. Morphologically normal somatic embryos were obtained on culture media composed of 20.70 or 41.41 μM picloram (PIC) with the addition of 0.044 μM 6-benzylaminopurine (BA), resulting in a 33 and 24% of conversion into plants, respectively. The source of explants and the developmental stage of the leaves had a marked effect on somatic embryogenesis. The second folded immature leaves from in vitro growing plants were the most responsive producing up to 30% embryogenesis in MS+41.41 μM PIC. Embryos converted into plants after transfer to MS medium devoid of growth regulators and these plants were successfully acclimatised. Adventitious shoots were obtained on culture media supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) or naphthaleneacetic acid (NAA) with or without 0.044 μM BA, achieving plant regeneration in the induction media. The highest percentage of bud formation was obtained on culture medium composed of␣MS+10.74 μM NAA+0.044 μM BA (12.5%). Roots were formed on all culture media tested. Regenerated plants were transferred to pots and grew well under greenhouse conditions. 相似文献
6.
C. Pugliesi M. Rabaglio F. Cecconi S. Baroncelli 《Plant Cell, Tissue and Organ Culture》1992,28(2):125-128
Different plant explants of Persian buttercup (Ranunculus asiaticus L.) were screened for callus induction and adventitious shoot regeneration on different media to establish totipotent cultures. Murashige & Skoog (MS) medium was used, supplemented with different concentrations of the following growth regulators: kinetin, benzyladenine (BA), naphthaleneacetic acid (NAA) and indoleacetic acid (IAA). Callus was induced and adventitious buds regenerated only from cotyledonary explants after 4–5 weeks. Subculture of the regenerated buds on the same basal medium in presence of gibberellic acid (GA3) and BA produced well-organized shoots. Rooting was obtained by transferring shoots to growth regulator-free MS medium. A high rate of shoot multiplication has been achieved on medium with high concentration of kinetin and long-day photoperiod. Finally the plants were successfully transferred to soil and grown in a greenhouse. 相似文献
7.
Direct plant regeneration from leaf explants of Plumbago species and its genetic fidelity through RAPD markers 总被引:1,自引:0,他引:1
G R ROUT 《The Annals of applied biology》2002,140(3):305-313
In vitro plant regeneration was achieved from leaf explants of Plumbago rosea and Plumbago zeylanica on Murashige & Skoog (1962) medium supplemented with 1.5 mg litre?1 6‐benzylaminopurine, 0.25 mg litre?1 indole‐3‐acetic acid, 50 mg litre?1 adenine sulfate and 3% (w/v) sucrose. The shoot initials developed within 2–3 wk on the leaf margin as well as from the wounds of the leaf. High frequency shoot‐bud regeneration was achieved on similar medium in subsequent subcultures. The semi‐mature leaves produced more shoot‐buds as compared to the younger leaves. Mature leaves did not show any response for shoot bud initiation. More than 85% of the semi‐mature explants produced shoot‐buds per leaf explant within 4 wk of culture. Shoots rooted easily on medium having half‐strength basal Murashige & Skoog (1962) medium supplemented with 0.25 mg litre?1 indole‐3‐butyric acid and 2% (w/v) sucrose; 84–92% of the in vitro rooted plantlets survived in the greenhouse. The regenerated plantlets appeared morphologically similar to the mother plants. No variation was detected among the regenerated plants by the use of Randomly Amplified Polymorphic DNA (RAPD) markers. This method might be useful for assessing plant improvement programmes. 相似文献
8.
A tissue culture method is described for clonal multiplication of Leucaena leucocephala K67 using single lateral bud explants from 2–3 m tall greenhouse grown trees. N-6 benzyladenine (BA: 3.0 mg.1-1) and napthaleneacetic acid (NAA: 0.05 mg.1-1) in Murashige & Skoog's (MS) medium were found to be best suited for multiple shoot differentiation in 4–5 week old cultures.
Analysis of variance of the main treatment effects of BA and NAA on shoot parameters showed that BA significantly (P=0.001) affected shoot development while NAA did not. A shoot multiplication rate of 22±3.63 shoots per bud explant was obtained
in 150 days on 1/2 strength MS medium with 3.0 mg.1-1 BA and 0.05 mg.1-1 NAA. Shoots developed adventitious roots within 15 days in 1/2 strength MS medium containing indole-3-butyric acid (IBA:
3.0 mg.1-1) and Kinetin (0.05 mg.1-1). Eighty percent of the transplanted plantlets are being grown in greenhouse conditions. 相似文献
9.
Hai Ping Hong Hongyi Zhang Paula Olhoft Steve Hill Hunt Wiley Effie Toren Helke Hillebrand Todd Jones Ming Cheng 《In vitro cellular & developmental biology. Plant》2007,43(6):558-568
A regeneration and transformation system has been developed using organogenic calluses derived from soybean axillary nodes
as the starting explants. Leaf-node or cotyledonary-node explants were prepared from 7 to 8-d-old seedlings. Callus was induced
on medium containing either Murashige and Skoog (MS) salts or modified Finer and Nagasawa (FNL) salts and B5 vitamins with
various concentrations of benzylamino purine (BA) and thidiazuron (TDZ). The combination of BA and TDZ had a synergistic effect
on callus induction. Shoot differentiation from the callus occurred once the callus was transferred to medium containing a
low concentration of BA. Subsequently, shoots were elongated on medium containing indole-3-acetic acid (IAA), zeatin riboside,
and gibberellic acid (GA). Plant regeneration from callus occurred 90 ∼ 120 d after the callus was cultured on shoot induction
medium. Both the primary callus and the proliferated callus were used as explants for Agrobacterium-mediated transformation. The calluses were inoculated with A. tumefaciens harboring a binary vector with the bar gene as the selectable marker gene and the gusINT gene for GUS expression. Usually 60–100% of the callus showed transient GUS expression 5 d after inoculation. Infected
calluses were then selected on media amended with various concentrations of glufosinate. Transgenic soybean plants have been
regenerated and established in the greenhouse. GUS expression was exhibited in various tissues and plant organs, including
leaf, stem, and roots. Southern and T1 plant segregation analysis of transgenic events showed that transgenes were integrated into the soybean genome with a copy
number ranging from 1–5 copies. 相似文献
10.
Imre A. Tamas Janet L. Schlossberg-Jacobs Robert Lim Leslie B. Friedman Charles C. Barone 《Plant Growth Regulation》1989,8(2):165-183
The hormonal control of axillary bud growth was investigated in cultured stem segments of Phaseolus vulgaris L. When the stem explants were excised and implanted with their apical end in a solid nutrient medium, outgrowth of the axillary buds-located at the midline of the segment-was induced. However, if indoleacetic acid (IAA) or naphthaleneacetic acid (NAA) was included in the medium, bud growth was inhibited. The exposure of the apical end to IAA also caused bud abscission and prevented the appearance of new lateral buds.In contrast to apically inserted segments, those implanted in the control medium with their basal end showed much less bud growth. In these segments, the auxin added to the medium either had no effect or caused a slight stimulation of bud growth.The IAA transport inhibitor N-1-naphthylphthalamic acid (NPA) relieved bud growth inhibition by IAA. This suggests that the effect of IAA applied at the apical end requires the transport of IAA itself rather than a second factor. With the apical end of the segment inserted into the IAA-containing medium, simultaneous basal application of IAA relieved to some extent the inhibitory effect of the apical IAA treatment. These results, together with data presented in a related article [Lim R and Tamas I (1989) Plant Growth Regul 8: 151–164], show that the polarity of IAA transport is a critical factor in the control of axillary bud growth.Of the IAA conjugates tested for their effect on axillary bud growth, indoleacetyl alanine, indoleacetic acid ethyl ester, indoleacetyl-myo-inositol and indoleacetyl glucopyranose were strongly inhibitory when they were applied to the apical end of the stem explants. There was a modest reduction of growth by indoleacetyl glycine and indoleacetyl phenylalanine. Indoleacetyl aspartic acid and indoleglyoxylic acid had no effect.In addition to IAA and its conjugates, a number of other plant growth substances also affected axillary bud growth when applied to the apical end of stem segments. Myo-inositol caused some increase in the rate of growth, but it slightly enhanced the inhibitory effect of IAA when the two substances were added together. Gibberellic acid (GA3) caused some stimulation of bud growth when the explants were from younger, rather than older plants. The presence of abscisic acid (ABA) in the medium had no effect on axillary bud growth. Both kinetin and zeatin caused some inhibition of axillary buds from younger plants but had the opposite effect on buds from older ones. Kinetin also enhanced the inhibitory effect of IAA when the two were applied together.In conclusion, axillary buds of cultured stem segments showed great sensitivity to auxins and certain other substances. Their growth responded to polarity effects and the interaction among different substances. Therefore, the use of cultured stem segments seems to offer a convenient, sensitive and versatile test system for the study of axillary bud growth regulation. 相似文献
11.
An efficient protocol is described for the rapid in vitro multiplication of an endangered medicinal plant, Tylophora indica (Burm. f.) Merrill, via enhanced axillary bud proliferation from nodal explants collected from young shoots of a two-year-old
plant. The physiological effects of growth regulators [6-benzyladenine (BA), kinetin (Kin) thidiazuron (TDZ), indole-3-acetic
acid (IAA), indole-3-butyric acid (IBA) or α-naphthalene acetic acid (NAA)], ascorbic acid (AA), different strengths of Murashige
and Skoog (MS) medium and various pH levels on in vitro morphogenesis were investigated. The highest number (8.6 ± 0.71) of
shoots and the maximum average shoot length (5.2 ± 0.31 cm) were recorded on MS medium supplemented with 2.5 μM BA, 0.5 μM
NAA and 100 mg/l AA at pH 5.8. Rooting was best achieved on half-strength MS medium augmented with 0.5 μM IBA. The plantlets
regenerated in vitro with well-developed shoot and roots were successfully established in pots containing garden soil and
grown in a greenhouse with a 90% survival rate. The regenerated plants did not show any immediate detectable phenotypic variation.
The described method can be successfully employed for large-scale multiplication and long-term in vitro conservation of T. indica. 相似文献
12.
In Chrysanthemum leaf explants cultivated in vitro the capacity to covalently link polyamines to protein substances exists. This plant enzyme activity shows some similarities with mammalian transglutaminases. In foliar explants cultured on a medium promoting bud or root formation increasing levels of transglutaminase-like activity occurred during the first days of culture when cell multiplication was rapid then the levels declined as the rate of cell division decreased and differentiation occurred. Undifferentiated callus exhibited low transglutaminase-like activity. Transglutaminase-like activity also increased in rapidly proliferating and growing organs (roots and buds initiated from the foliar explants) and decreased during maturity. The relationship among transglutaminases-like activity, cell division, bud and root formation is discussed.Abbreviations TGase
transglutaminase
- BA
benzyladenine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- Put
putrescine
- Spd
spermidine 相似文献
13.
Organogenic callus cultures of Solanum paludosum were obtained from root, hypocotyle and cotyledon explants of plantlets cultured in sterile conditions. These callus cultures developed multiple shoots which proliferated in Murashige and Skoog basal liquid medium. These multiple shoots produced solamargine, the main steroidal glycoalkaloid present in the unripe fruits.The optimization of the macronutrient composition of the liquid medium was performed by a method derived from the plant composition. This approach results in the establishment of an appropriate medium (SPOM medium) suitable for the improvement of both growth and solamargine production by multiple shoot cultures of S. paludosum. 相似文献
14.
Liliana Iturriaga Miguel Jordan Carlos Roveraro Andrée Goreux 《Plant Cell, Tissue and Organ Culture》1994,37(2):201-204
Sophora toromiro (Phil.) Skottsb. is a woody species in imminent danger of extinetion. Since natural regeneration by seeds is poor and plant growth is very slow, asexual propagation is necessary. In vitro regeneration from 3- to 4-month-old seedlings was achieved. A range of naphthaleneacetic acid and benzyladenine concentrations induced root formation in nodal segment explants, developing plantlets, and also promoted axillary bud development. In subculture, nodal sections derived from axillary growth initiated multiple shoot formation and roots in a liquid medium leading to plantlet formation.Abbreviations NAA
naphthaleneacetic acid
- BA
benzyladenine
- IBA
indolebutyric acid
- GA3
gibberellic acid 相似文献
15.
B. Wang D. X. Peng Z. X. Sun N. Zhang S. M. Gao 《In vitro cellular & developmental biology. Plant》2008,44(2):105-111
Ramie [Boehmeria nivea (L.) Gaud] is one of the most important perennial fiber crops in China. In vitro tissue culture of ramie could serve as an important means for its improvement through genetic transformation. To improve
the regeneration capacity of ramie, the effects on plant regeneration of donor plant age, basal medium, plant growth regulators,
and culture conditions were evaluated using explants derived from the cotyledon, hypocotyl, leaf, petiole, and stem of ramie
seedlings. Cotyledons and hypocotyls excised from 4-d-old seedlings and leaves and petioles and stems from 15-d-old seedlings
were optimal explants. The highest regeneration efficiency was obtained on Murashige and Skoog salts with Gamborg’s B5 vitamins basal medium containing 2.27 μM thidiazuron (TDZ) and 0.054 μM naphthaleneacetic acid (NAA) for the five explant
types tested. A photoperiod of 16:8 h (light/dark) was found to be superior than continuous darkness for regeneration of ramie
using TDZ. The regenerated shoots were transferred to hormone-free medium for shoot elongation and successfully rooted on
half-strength Murashige and Skoog supplemented with 0.134 μM NAA. The rooted plantlets with four to five leaves were transplanted
to greenhouse for further growth. 相似文献
16.
High-frequency shoot multiplication in<Emphasis Type="Italic"> Curcuma longa</Emphasis> L. using thidiazuron 总被引:2,自引:0,他引:2
Prathanturarug S Soonthornchareonnon N Chuakul W Phaidee Y Saralamp P 《Plant cell reports》2003,21(11):1054-1059
The effects of plant growth regulators, explant types, and culture regimens were investigated on in vitro shoot proliferation from terminal bud explants of Curcuma longa. Each bud was longitudinally divided into four equal pieces, each 1 cm in length, and used as explants. These were then cultured on MS medium supplemented with 18.17 microM thidiazuron for 4 weeks prior to transfer to MS medium without growth regulator for 8 weeks. Under these conditions, a shoot induction rate of 18.22+/-0.62 shoots/explant was obtained after 12 weeks of cultures. Spontaneous rooting was achieved. The regenerated plants were transferred to soil under greenhouse conditions and subsequently grown successfully in the field. 相似文献
17.
Camille L. Hyde Gregory C. Phillips 《In vitro cellular & developmental biology. Plant》1996,32(2):72-80
Summary Chile pepper (Capsicum annuum L.) plants were regenerated from cotyledon explantsin vitro in four major stages: bud induction, bud enlargement, shoot elongation, and root development. Bud induction medium contained
0.5 mg/L (2.9μM) indole-3-acetic acid and 2 mg/L (8.9 μM) N6-benzyladenine. Bud enlargement occurred, and an occasional shoot appeared when medium with 2 mg/L (6μM) gibberellic acid, 2 mg/L (8.9 μM) N6-benzyladenine, and 5 mg/L (29.4 μM) silver nitrate was used. Most shoots elongated after placement on a third medium without plant growth regulators or on fresh
plates of bud enlargement medium. Incubations were for 2, 2, and 4 weeks, respectively, at 28.5°C and continuous light. Treatment
with silver nitrate was necessary for multiple shoot production and elongation to occur in the third culture stage and was
most effective when present in the second-stage medium but not in the bud induction medium. Sixteen to 26% of the shoots rooted
in medium with 1 mg/L (5.4 μM) 1-naphthaleneacetic acid after 1 month. Additional shoots transferred to a second rooting medium with 0.1 or 1.0 mg/L (0.54
or 5.4 μM) 1-naphthaleneacetic acid developed roots, increasing the overall rooting efficiency to 70–72%. Most rooted shoots grew well
and produced viable seeds when grown in the greenhouse. Other cytokinins tested for plant regeneration were zeatin and thidiazuron.
Zeatin induced few shoots and fewer well-developed plants. Thidiazuron induced multiple shoots 4 months after culture began,
but many were small and did not elongate further. Phytagar tissue culture grade proved superior to other agars tested, increasing
bud induction frequency from 0-33% to 80–93% and eliminating explant hyperhydricity. 相似文献
18.
Summary A protocol for large-scale propagation of Phragmites communis Trin. by adventitious bud formation and plant regeneration was established. Adventitious buds were induced through either
the indirect pathway or the direct pathway from stem explants of Phragmites communis. In the indirect pathway, it was essential to decrease the level of 2,4-dichlorophenoxyacetic acid from 9.1 to 0.5 μM to induce adventitious buds and achieve plant regeneration. In the direct pathway, the effects of different benzylaminopurine
(BA) concentrations in the medium, and different positions of the explants, on adventitious bud formation were determined.
Murashige and Skoog (MS) medium supplemented with 5.4μM α-naphthaleneacetic acid (NAA) and 53.4 μM BA, and the bottom part of stem explants were most responsive for the differentiation of adventitious shoot buds. The highest
differentiation frequency was 20–30 adventitious shoot buds per stem node tissue. Elongation and proliferation of adventitious
buds were achieved on MS medium supplemented with 13.3 μM BA and 5.4 μM NAA. Shoots were rooted in liquid half-strength MS medium with 5.4 μM NAA+4.9 μM indole-3-butyric acid. Rooted plants survived (87.5%) and grew well after transfer into soil for 4 wk. More than 20 000 regenerated
plants of a salt-tolerant variant line of Phragmites communis have been produced. This protocol is useful for clonal micropropagation and possibly for Agrobacterium- mediated gene transfer in P. communis. 相似文献
19.
Shyamkumar Barampuram Byung Yeoup Chung Seung Sik Lee Byung Chull An Eun Mi Lee Jae-Young Cho 《In vitro cellular & developmental biology. Plant》2009,45(2):155-161
The present study demonstrates the establishment of embryogenic tissue from seeds and (seedling-derived hypocotyls) shoot
base explants derived from seedlings of Eremochloa ophiuroides. The highest percentage of callus induction obtained from seed and young shoot base explants was 52.0% and 66.6% on Murashige
and Skoog (MS) basal media supplemented with 9.0 μM and 18.1 μM 2,4-dichlorophenoxyacetic acid (2,4-D), respectively. The
type of callus obtained from both types of explants was off-white to yellow in color and non-friable and shiny in texture.
Excised callus from the explants was subcultured onto fresh media of the same recipe for further proliferation. After 10–12 d
of subculture, a yellow, globular, friable embryogenic callus was obtained from the initial callus. The highest percentage
of embryogenic calli obtained at 40.0% was observed on media containing 2.2 μM 2,4-D. The highest regeneration rate of 46.6%
was observed on MS media supplemented with 0.4 μM 2,4-D and 2.2 μM benzylaminopurine (BA). Regenerated shoots were rooted
in MS basal medium. Plants with well-developed roots were transferred to pots containing a soil mix and acclimatized in greenhouse
conditions. Four weeks post-transfer, acclimatized plants showed 100% survival and remained healthy and green. This is the
first report of a successful method for induction of somatic embryogenesis with subsequent plant regeneration in centipede
grass and demonstrates the establishment of embryogenic callus and efficient plant regeneration with potential application
in the development of genetic transformation systems for centipede grass. 相似文献
20.
Jin Cui Jianjun Chen Richard J. Henny 《In vitro cellular & developmental biology. Plant》2009,45(1):34-43
Plant regeneration through direct somatic embryogenesis in Aeschynanthus radicans ‘Mona Lisa’ was achieved in this study. Globular somatic embryos were formed directly from cut edges of leaf explants and
cut ends or on the surface of stem explants 4 wk after culture on Murashige and Skoog (MS) medium supplemented with N-phenyl-N′-1, 2, 3-thiadiazol-5-ylurea (TDZ) with α-naphthalene acetic acid (NAA), TDZ with 2,4-dichlorophenoxyacetic acid (2,4-D),
or 6-benzylaminopurine (BA) or kintin (KN) with 2,4-D. MS medium containing 9.08 μM TDZ and 2.68 μM 2,4-D resulted in 71%
of stem explants producing somatic embryos. In contrast, 40% of leaf explants produced somatic embryos when induced in medium
containing 6.81 μM TDZ and 2.68 μM 2,4-D. Somatic embryos matured, and some germinated into small plants on the initial induction
medium. Up to 64% of stem explants cultured on medium supplemented with 9.08 μM TDZ + 2.68 μM 2,4-D, 36% of leaf explants
cultured on medium containing 6.81 μM TDZ and 2.68 μM 2,4-D had somatic embryo germination before or after transferring onto
MS medium containing 8.88 μM BA and 1.07 μM NAA. Shoots elongated better and roots developed well on MS medium without growth
regulators. Approximately 30–50 plantlets were regenerated from each stem or leaf explant. The regenerated plants grew vigorously
after transplanting to a soil-less substrate in a shaded greenhouse with more than a 98% survival rate. Three months after
their establishment in the shaded greenhouse, 500 plants regenerated from stem explants were morphologically evaluated, from
which five types of variants that had large, orbicular, elliptic, small, and lanceolate leaves were identified. Flow cytometry
analysis of the variants along with the parent showed that they all had one identical peak, indicating that the variant lines,
like the parent, were diploid. The mean nuclear DNA contents of the variant lines and their parent ranged from 4.90 to 4.99 pg
2C−1, which were not significantly different statistically. The results suggest that the regenerated plants have a stable ploidy
level, and the regeneration method established in this study can be used for rapid propagation of ploidy-stable Aeschynanthus radicans. 相似文献