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1.
Pueraria tuberosa, a medicinally important leguminous plant, yielding various isoflavanones including puerarin, is threatened, thus requiring
conservation. In this study, fresh shoot sprouts of P. tuberosa, produced by tubers, were used as explants for in vitro micropropagation. Surface-sterilized nodal shoots were incubated
on Murashige and Skoog (MS) medium supplemented with 8.88 μM benzyladenine (BA), 50 mg l−1 ascorbic acid, and 25 mg l−1 of each of citric acid and adenine sulphate. Cut ends of nodal stem segments rapidly turned brown, and cultures failed to
establish. When 100 mg l−1 ascorbic acid (ABA) and 25.0 mg l−1 polyvinyl pyrrolidone (PVP) were added to the medium, explants remained healthy, and cultures were established. Bud-breaking
of nodal stem explants resulted in multiple shoot formation. Shoots proliferated (35–40 shoots per culture vessel) on MS medium
as described above, but supplemented with 4.44 μM BA and 0.57 μM indole acetic acid (IAA) and additives. After 4–5 passages,
proliferating shoots exhibited tip-browning and decline in growth and multiplication. However, when shoots were transferred
to fresh shoot proliferation medium supplemented with 2.32 μM kinetin (Kn), sustained growth and high rate of shoot proliferation
(50–60 shoots per culture vessel) was observed. Shoots rooted when transferred to medium consisting of half- strength MS medium
with 9.84 μM indole butyric acid (IBA) and 0.02% activated charcoal. Alternatively, individual shoots were pulsed with 984.0 μM
IBA and transferred to glass bottles containing sterile and moistened soilrite. These shoots rooted ex-vitro and were acclimatized
in the greenhouse. Plants were then analyzed for puerarin content using HPLC, and leaves showed maximum accumulation of purerarin. 相似文献
2.
Pterocarpus marsupium (Bijasal) is a valuable multipurpose forest tree. The regeneration rate in natural habitat is low and the tree is overexploited.
An in vitro propagation protocol has been developed from nodal explants obtained from in vitro raised 18-day-old axenic seedlings.
The highest shoot regeneration frequency (85%), maximum number of multiple shoots (8.6) as well as length (4.8 cm) were induced
from nodal explants on Murashige and Skoog (MS) medium amended with 4.0 μM 6-benzyladenine (BA), 0.5 μM indole-3-acetic acid
(IAA) and 20 μM adenine sulphate (AdS). The percentage of shoot multiplication as well as the number of shoots per node remained
the same during the first two subculture, afterwards a decline was recorded. Rooting was best induced in microshoots excised
from proliferated shoot cultures on semisolid hormone-free half-strength MS medium, after a pulse (dip) treatment for 7 days
in half-strength MS liquid medium containing 100 μM indole-3-butyric acid (IBA) and 15.84 μM phloroglucinol (PG). The in vitro-raised
plantlets were potted and acclimatized under culture room conditions for 4 weeks before their transfer to a greenhouse, where
the established plants showed 75% survival. 相似文献
3.
Padar (Stereospermum personatum, family Bignoniaceae) is a well-known medicinal tree. Its complete regeneration occurred through shoot bud culture in vitro. The seeds germinated
sequentially on plastic trays and polyethylene bags for 21 days served as explants source. Nodal segments from the seedlings
were established on MS medium supplemented with 4.44 μM BA, in which 86.6% nodes showed shoot bud elongation. Then, nodal
segments from the developed shoots were cultured on MS medium with several BA concentrations; best shoot multiplication was
obtained with 0.44 μM BA. In a second experiment where PVP was added to proliferation medium, nodal segments from developed
shoots produced maximum 2.78 shoots per node. The nodal segments showed shoot multiplication up to seventh subculture on.
Finally, shoots were rooted on MS medium with 2.46 μM IBA. The plants transferred to net pots containing coco-peat were acclimatized
in green house, where more than 80% plants survived and grew normally. 相似文献
4.
An efficient, rapid and large scale propagation of a multipurpose herb, Ocimum basilicum through in vitro culture of nodal segments with axillary buds from mature plants has been accomplished. Among the cytokinins,
6-benzyladenine (BA), thidiazuron (TDZ), kinetin (Kin) and 2-isopentenyl adenine (2-iP) tested as supplements to Murashige
and Skoog (MS) medium, 5.0 μM BA was optimum in inducing bud break. The highest rate of shoot multiplication was achieved
on half-strength MS medium supplemented with 2.5 μM BA and 0.5 μM indole-3-acetic acid (IAA) combination. The shoots regenerated
from TDZ supplemented medium when subcultured to hormone-free MS medium considerably increased the rate of shoot multiplication
and shoot length by the end of third subculture. For rooting, MS medium supplemented with 1.0 μM indole-3-butyric acid (IBA)
proved to be better than that supplemented with IAA or α-naphthalene acetic acid (NAA). The in vitro raised plantlets with
well developed shoots and roots were successfully established in earthen pots containing garden soil and were grown in greenhouse
with 90% survival rate. Chlorophyll a and b, carotenoids and net photosynthetic rate were measured in leaves during ex vitro
acclimatization at 0, 7, 14, 21 and 28 days. Firstly these parameters showed a decreasing trend but subsequently increased
after 7 days of acclimatization. These findings indicate that the adaptation of micropropagated plants to ex vitro conditions
is more extended in time than generally accepted. 相似文献
5.
In vitro propagation of northern red oak (Quercus rubra) shoots was successful from cotyledonary node explants excised from 8-wk-old in vitro grown seedlings. Initially, four shoots per explant were obtained on Murashige and Skoog (MS) medium supplemented with 4.4 μM
6-benzylaminopurine (BA), 0.45 μM thidiazuron (TDZ), and 500 mg l−1 casein hydrolysate (CH) with a regeneration frequency of 64.7% after 3 wk. Subculturing explants (after harvesting shoots)
to fresh treatment medium significantly increased shoot bud regeneration (16.6 buds per explant), but the buds failed to develop
into shoots. A higher percentage (73.3%) of the explants regenerated four shoots per explant on woody plant medium (WPM) supplemented
with 4.4 μM BA, 0.29 μM gibberellic acid (GA3), and 500 mg l−1 CH after 3 wk. Explants subcultured to fresh treatment medium after harvesting shoots significantly increased shoot regeneration
(16 shoots per explant). Shoot elongation was achieved (4 cm) when shoots were excised and cultured on WPM supplemented with
0.44 μM BA and 0.29 μM GA3. In vitro regenerated shoots were rooted on WPM supplemented with 4.9 μM indole-3-butyric acid. A higher percentage regeneration response
and shoot numbers per explant were recorded on WPM supplemented with BA and GA3, than on MS medium containing BA and TDZ. Lower concentrations of BA and GA3 were required for shoot elongation and prevention of shoot tip necrosis. Each cotyledonary node yielded approximately 20
shoots within 12 wk. Rooted plantlets were successfully acclimatized. 相似文献
6.
An efficient, rapid and reproducible plant regeneration protocol was successfully developed for Cassia angustifolia using nodal explants excised from 14-day-old aseptic seedlings. Of the two cytokinins, 6-benzyladenine (BA) and thidiazuron
(TDZ) evaluated as supplements to Murashige and Skoog (MS) medium, TDZ at an optimal concentration of 5.0 μM was effective
in inducing multiple shoots. The highest rate of shoot multiplication was achieved on MS medium supplemented with 5.0 μM TDZ
and 1.0 μM indole-3-acetic acid (IAA) at pH 5.8. The regenerated shoots when subcultured on hormone free MS medium considerably
increased the rate of shoot multiplication and shoot length by end of fourth subculture passage. Rooting was achieved on the
isolated shoots using MS medium with 60 μM indole -3- butyric acid (IBA) and 1% activated charcoal for 1 week and subsequently
transferring the shootlets to half strength MS liquid media without IBA and activated charcoal. The in vitro raised plantlets
with well-developed shoot and roots were successfully established in earthen pots containing garden soil and grown in greenhouse. 相似文献
7.
A simple, rapid and efficient protocol for micropropagation of Cardiospermum halicacabum via axillary bud multiplication has been successfully developed. The organogenic competence of nodal segments was investigated
on Murashige and Skoog (MS) medium supplemented with different concentrations of benzyladenine (BA), kinetin (Kn), thidiazuron
(TDZ) and 2-isopentenyladenine (2-iP). Multiple shoots differentiated directly without callus mediation within 4 weeks when
explants were cultured on a medium fortified with cytokinins. The maximum number of shoots (14.83 ± 0.52) was developed on
a medium supplemented with 0.3 μM TDZ. Such proliferating shoots when subcultured onto MS media devoid of TDZ gave the highest
rate of shoot multiplication (35.66 ± 1.00) by the end of fourth subculture passage. Elongated shoots were rooted on 1/3 MS
medium augmented with 0.5 μM IAA. The plantlets thus obtained were successfully hardened and transferred to greenhouse. 相似文献
8.
Summary A viable protocol has been developed for direct and indirect shoot regeneration of Vernonia cinerea. To establish a stable and high-frequency plant regeneration system, leaf and stem explants were tested with different combinations
of α-naphthalene acetic acid (NAA), indole-3-acetic acid (IAA), and benzylaminopurine (BA). Lateral buds on nodal explants
grew into shoots within 2 wk of culture in Murashige and Skoog (MS) basal medium supplemented with 20.9 μM BA. Excision and culture of nodal segments from in vitro-raised shoots on fresh medium with the same concentration of BA facilitated development of more than 15 shoots per node.
Similarly leaf, nodal, and internodal explants were cultured on MS basal medium supplemented with different concentrations
of BA, NAA, and IAA either alone or in combinations for callus induction and organogenesis. Shoot buds and/or roots were regenerated
on callus. Shoot buds formed multiple shoots within 4 wk after incubation in induction medium. Adventitious buds and shoots
proliferated when callus was cut into pieces and subcultured on MS basal medium containing 20.9 μM BA and 5.3 μM NAA. This combination proved to be the best medium for enhanced adventitious shoot bud multiplication, generating a maximum
of 50 shoots in 4 wk. This medium was also used successfully for shoot proliferation in liquid medium. Root formation was
observed from callus induced in medium containing 8.05–13.4 μM NAA. Regenerated shoots exhibited flowering and root formation in MS basal medium without any growth regulators. Plantlets
established in the field showed 85% survival and exhibited identical morphological characteristics as the donor plant. 相似文献
9.
Ganeshan Sivanandhan Thankaraj Salammal Mariashibu Muthukrishnan Arun Manoharan Rajesh Sampath Kasthurirengan Natesan Selvaraj Andy Ganapathi 《Acta Physiologiae Plantarum》2011,33(6):2279-2288
An efficient mass multiplication protocol was developed for Withania somnifera (L.) Dunal from nodal explants of field-grown plants on Murashige and Skoog medium (MS) supplemented with 6-benzyladenine
(BA) [1.5 mg L−l], indole-3-acetic acid (IAA) [0.3 mg L−l] and with the addition of polyamine, spermidine (20 mg L−l) (shoot multiplication medium). A total of 46.4 shoots were obtained from nodal explants and they were elongated in the same
medium in a culture duration of 6 weeks. The elongated shoots produced roots in MS medium fortified with putrescine (20 mg L−l) after 4 weeks, and all the rooted plants were successfully hardened and acclimatized with a survival rate of 100%. An average
of 276 shoots (46 × 6) was produced when at least six nodal explants obtained from each of the 46 in vitro grown shoots were
cultured by microcutting method in the same shoot multiplication medium. On an average, 12,696 plants could be produced from
all the shoots (276 × 46) by microcuttings in a period of 7 months. HPLC revealed a significant increase in the quantities
of withanolide A, withanolide B, withaferin A and withanone in the leaves, stems, and roots of in vitro regenerated plants
compared to the field-grown parent plants. Ploidy analysis using flow cytometry revealed genetic stability of in vitro regenerated
plants. This protocol will be useful for scale-up production of withanolides on commercial scale. 相似文献
10.
Conditions for plant regeneration from excised shoot tips of Vigna radiata were studied. Complete plants were regenerated directly without an intervening callus phase from shoot tips on basal medium (MS salts+B5vitamins). Regeneration frequency varied with genotype, explant size and growth regulator combinations in the medium. Addition of cytokinins induced a variable amount of callus at the base of the shoot tip, followed by multiple shoot formation. Benzyladenine (BA), kinetin and zeatin at 5×10-6 M each induced multiple shoots in 100% of the explants but the highest number of regenerants per explant (9) was produced with BA. The efficacy of BA for shoot multiplication was not improved when it was supplemented with naphthaleneacetic acid (NAA) or indoleacetic acid (IAA). NAA or adenine sulphate, when applied alone, induced complete plantlets. The growth regulator requirement of explants for the induction of multiple shoots varied with explant size. The shoot tip explants maintained proliferation ability on subculture. None of the treatments was effective in inducing shoot bud differentiation from callus. Regenerated shoots were rooted on MS basal medium and MS supplemented with either IAA or indolebutyric acid. The rooted plants were transferred to the field; 60% subsequently survived and grew.Abbreviations BM
basal medium [MS (Murashige & Skoog 1962) salts+B5 (Gamborg et al. 1968) vitamins]
- BA
6-benzyladenine
- AdS
adenine sulphate
- IAA
indole-3-acetic acid
- NAA-1
naphthaleneacetic acid
- IBA
indolebutyric acid 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.
Surya Prakash Johannes Van Staden 《In vitro cellular & developmental biology. Plant》2008,44(4):338-341
A method for in vitro regeneration of Searsia dentata from nodal and shoot tip explants derived from mature trees is outlined. Nodal explants produced multiple shoots from the
axis when cultured on Murashige and Skoog (MS) medium containing 3% sucrose supplemented with 0, 5, 7.5, 10, or 12.5 μM N
6-benzyladenine (BA). An average of 5.3 shoots was obtained from nodal explants on 10 μM BA. For shoot tip explants, however,
supplementation of α-naphthaleneacetic acid (NAA) with BA favored a caulogenic response. A maximum of 6.1 shoots were produced
per shoot tip explant on MS containing 7.5 μM BA plus 5.0 μM NAA. The in vitro-regenerated shoots produced roots when transferred
to full-strength MS medium containing 3% sucrose and 10 μM indole-3-butyric acid (IBA). The developed plantlets were transferred
initially to a mist house. After an initial acclimatization period of 3–4 mo, plantlets were shifted to the greenhouse where
they thrived for 9 mo. The standardized protocol for mass propagation of S. dentata should eliminate the dependence on natural stands of plants for traditional medicinal purposes, and will also serve as a
means of conservation as the species is heavily overexploited. 相似文献
13.
Bambusa balcooa is one of the most commercially important bamboo species. Regeneration of this species by sexual means is impossible because
no seeds are set after flowering. Vegetative propagation is hindered due to bulky propagules, low rooting ability of the culm
and branch cuttings, and seasonal specificity. This makes in vitro-based methods of regeneration important. This paper describes an efficient micropropagation protocol for multiplication of
B. balcooa from nodal explants. Nodal segments were surface sterilized with 0.1% mercuric chloride for 10 min, and cultured on Murashige
and Skoog (MS) medium supplemented with 4.4 μM 6-benzylaminopurine (BAP), 2.32 μM kinetin (Kn), and gelled with 0.2% w/v gelrite. Eighty-five percent of explants could be established in vitro with 90% of these achieving bud break. In vitro-formed shoots were successfully multiplied in MS liquid medium supplemented with 6.6 μM BAP, 2.32 μM Kn, 2.5% v/v coconut water, and 100 mg l−1
myo-inositol. Subculturing shoots every 3 wk yielded a consistent proliferation rate of 4.11-fold without decline in vigor. Shoot
clusters, containing 5 to 8 shoots, were rooted with 87.5% success in 1/2 MS supplemented with 5.71 μM indole-3-acetic acid
(IAA), 4.9 μM indole-3-butyric acid (IBA), and 5.37 μM naphthaleneacetic acid (NAA) within 3 wk. Plants regenerated in this
manner were acclimatized in the greenhouse and under a shade net with 88% success. 相似文献
14.
Giovanni Iapichino Marcello Airò 《In vitro cellular & developmental biology. Plant》2008,44(4):330-337
Multiple shoots were induced on stem segments of an 8-y-old plant of Metrosideros excelsa Sol ex Gaertn. “Parnel”. Axillary shoots produced on uncontaminated explants were excised, segmented, and recultured in the
same medium to increase the stock of shoot cultures. The Murashige and Skoog (MS) medium, augmented with different concentrations
of 2- isopenthenyladenine (2iP) and indole-3-acetic acid (IAA), either singly or in combinations, as potential medium for
shoot multiplication by nodal segments was tested. In the following experiment, equal molar concentrations of four cytokinins
[2iP, kinetin, zeatin, and N
6-benzyladenine (BA)] in combination with equal molar concentrations of three auxins [IAA, α-naphthaleneacetic acid (NAA),
and indole-3-butyric acid (IBA)] were tested for ability to induce axillary shoot development from single-node stem segments.
The highest rate of axillary shoot proliferation was induced on MS agar medium supplemented with 1.96μM 2iP and 1.14μM IAA
after 6 wk in culture. Different auxins (IAA, IBA, and NAA) were tested to determine the optimum conditions for in vitro rooting of microshoots. The best results were accomplished with IAA at 5.71μM (89% rooting) and with IBA at 2.85 or 5.71μM
(86% and 86% rooting, respectively). Seventy and 90 percent of the microshoots were rooted ex vitro in bottom-heated bench (22 ± 2°C) after 2 and 4 wk, respectively. In vitro and ex vitro rooted plantlets were successfully established in soil. 相似文献
15.
Mehmet Nuri Nas Leyla Gokbunar Nevzat Sevgin Murat Aydemir Merve Dagli Zahide Susluoglu 《Plant Growth Regulation》2012,67(1):57-63
The effects of culture media and cytokinin types on micropropagation of mature Crataegus aronia L. were investigated. Using single-axillary bud explants, the growth of cultures on MS, WPM, DKW and NRM containing 4.44 μM
benzyladenine (BA) plus 0.05 μM indole-3-butyric acid (IBA), and on NRM containing thidiazuron, meta-Topolin (mT) or BA at
1.25, 2.5, 5.0 or 7.5 μM plus 0.05 μM IBA were compared. The culture medium had significant effects on shoot number and length.
In comparison with MS, DKW and WPM, shoot production was greater on NRM (5.7 shoots per explant). Shoot production on MS,
DKW and WPM (4.2, 4.2 and 4.1, respectively) were statistically similar to each other. Thidiazuron was detrimental to shoot
formation and caused formation of rosette shoots and/or large callus to form on explants. In the presence of mT, only some
of the explants developed into shoots. Benzyladenine was the only cytokinin that promoted both shoot proliferation and shoot
elongation. Higher shoot numbers were obtained at 5.0 and 7.5 μM BA compared to lower concentrations of BA. Over 80% of microshoots
rooted and rooted shoots were successfully acclimatized to ex vitro conditions. 相似文献
16.
Phillip A. Wadl Adam J. Dattilo Lisa M. Vito Robert N. Trigiano 《Plant Cell, Tissue and Organ Culture》2011,106(3):513-516
Pityopsis ruthii is an endangered herbaceous perennial species from the United States. In vitro multiplication of this species can be valuable
for germplasm conservation. Flower receptacles of P. ruthii were cultured on Murashige and Skoog medium (MS) supplemented with 11.4 μM indole-3-acetic acid (IAA) in combination with
2.2, 4.4 or 8.8 μM 6-benzyladenine (BA). Shoots were visible within 14–28 days and three plants were successfully rooted on
MS medium supplemented with 5.7 μM IAA. A two tailed t-test for paired-variates revealed that shoot regeneration on MS medium
amended with 11.4 μM IAA and 2.2 μM BA was significantly higher (P < 0.05) than on other treatments. Leaf explants were also cultured on MS not supplemented with growth regulators or supplemented
with 11.4 μM IAA in combination with 0, 2.2, 4.4 or 8.8 μM BA. Shoots were visible within 21–35 days and one plant was successfully
rooted on MS medium supplemented with 5.4 μM NAA. Shoot regeneration on MS medium augmented with 11.4 μM IAA and 2.2 μM BA
was significantly higher (P < 0.05) than the other treatments according to analysis of variance (ANOVA) with a rank transformation. Hyperhydricity and
rooting of shoots was problematic for explants derived from flower receptacles and leaf tissue, but viable plants were regenerated
using both explants sources indicating the potential role for micropropagation in the ex situ conservation of the species. 相似文献
17.
A rapid and highly-effective method for micropropagation from nodal segment and shoot tip explants was established for Coleus blumei Benth. Nodal segments and shoot tips were inoculated on MS medium containing 0.7 % agar, 3 % commercial sugar, and different
combinations of 6-benzyladenine (BA) with indole-3-acetic acid (IAA), indole-3-butyric acid (IBA) or α-naphthaleneacetic acid
(NAA). Hundred percent shoot induction from both explants was achieved on the medium containing BA (2 mg dm−3) and NAA (1 mg dm−3). Shoot tips were proved to be the better explant in comparison to nodal segments in having high rate of shoot induction
and more number of shoots. The same media conditions were found suitable for shoot multiplication. Multiplied shoots rooted
best on MS medium supplemented with IBA (2 mg dm−3). Micropropagated plants were successfully established in soil after hardening, with 100 % survival rate. 相似文献
18.
A simple, high frequency, and reproducible method for plant regeneration through direct organogenesis from cotyledonary leaf
explants of Jatropha curcas was developed using Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ) or 6-benzyl
aminopurine (BAP). Medium containing TDZ has greater influence on regeneration as compared to BAP. The induced shoot buds
were transferred to MS medium containing 10 μM kinetin (Kn), 4.5 μM BAP, and 5.5 μM α-naphthaleneacetic acid (NAA) for shoot
proliferation. The proliferated shoots could be elongated on MS medium supplemented with different concentrations and combinations
of BAP, indole-3-acetic acid (IAA), NAA, and indole-3-butyric acid (IBA). MS medium with 2.25 μM BAP and 8.5 μM IAA was found
to be the best combination for shoot elongation. However, significant differences in plant regeneration and shoot elongation
were observed among the genotypes studied. Rooting was achieved when the basal cut end of elongated shoots were dipped in
half strength MS liquid medium containing different concentrations and combinations of IBA, IAA, and NAA for 4 days, followed
by transfer to growth regulators free half strength MS medium supplemented 0.25 mg l−1 activated charcoal. Elongated shoot treated with 15 μM IBA, 5.7 μM IAA, and 11 μM NAA resulted in highest percent rooting.
The rooted plants could be established in soil with more than 90% survival rate. The method developed may be useful in improvement
of J. curcas through genetic modification. 相似文献
19.
Summary We have developed a highly efficient two-stage protocol for induction of multiple shoots from single node in vitro shoot tip explants of Decalepis hamiltonii. It was found that phenylacetic acid (PAA) had a synergistic effect on shoot multiplication when treated with N6-benzyladenine (BA). This protocol used PAA for both multiple shoot induction from nodal explants, elongation of primary shoots,
and initiation of adventitious shoot formation from primary shoots. Murashige and Skoog medium containing BA (2.22–31.08 μM) and α-naphthaleneacetic acid (0.27–10.74 μM) or PAA (7.34–36.71 μM) was used to initiate shoot formation from nodal explants. The maximum number of shoots per culture was produced on a medium
containing 31.08 μM BA and 14.68 μM PAA, while the longest shoot length and nodes were obtained on medium containing 22.2 μM BA and 14.68 μM PAA. Shoots subcultured on MS medium containing 22.2 μM BA and 14.68 μM PAA elongated along with secondary shoot formation. The shoots were rooted on medium containing 9.7 μM indole-3-butyric acid. The plantlets were acclimatized in soil with an 80–90% survival rate under field conditions. 相似文献
20.
Guang-Zhe Lin Xiao-Mei Zhao Soon-Kwan Hong Yu-Ji Lian 《Plant Cell, Tissue and Organ Culture》2011,106(1):93-103
We have developed a system for the in vitro regeneration of pasqueflowers (Pulsatilla koreana Nakai). The system was based on somatic embryogenesis and shoot organogenesis. Over a growth period of 6 weeks, multiple
shoots were initiated from leaf, petiole, and pedicel explants on Murashige and Skoog (MS) medium containing 0.5 mg l−1 indole-3-acetic acid (IAA) and zeatin (Zn), kinetin (Kin), or 6-benzyladenine (BA). We achieved 100% of adventitious shoot
induced when petiole and pedicel explants were cultured on MS, 0.5–2.0 mg l−1 Zn, and 0.5 mg l−1 IAA. Somatic embryos developed from the explants and generated shoots on MS medium containing 0.25 mg l−1 Zn and 0.5 mg l−1 IAA. Globular and heart-shaped stages of somatic embryos were observed. Histological studies have revealed the stages of
development of somatic embryos. For propagation and growth, the regenerated shoots from organogenic or embryogenic calluses
were transferred to MS medium containing either (1) 1.5 mg l−1 Zn and 0.05 mg l−1 IAA or (2) 1.0 mg l−1 BA and 0.05 mg l−1 IAA. After the length of the shoots reached 3 cm, the shoots initiated by organogenesis as well as those initiated by somatic
embryogenesis were transferred to the root induction medium. After 2 months of culture in half-strength MS with 1.5 mg l−1 α-naphthalene acetic acid (NAA), the rooting ratio was 93%. Finally, the rooted plantlets were acclimatized in a mixture
of mountain soil and perlite. 相似文献