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1.
Qin-Mei Wang Feng-Zhan Gao Xiang Gao Fan-Yu Zou Xin Sui Meng Wang Yue-Jun Hui Li Wang 《Plant Cell, Tissue and Organ Culture》2012,109(2):191-200
An efficient in vitro micropropagation system for Clivia miniata Regel was developed using basal tissues of young petals and young ovaries as explants. For callus induction, explants were
incubated on Murashige and Skoog (MS) medium containing either 2.22 μM 6-benzyladenine (BA) and 4.52 μM 2,4-dichlorophenoxyacetic
acid (2,4-D) or 4.44 μM BA, 5.37 μM α-naphthaleneacetic acid (NAA), and 9.05 μM 2,4-D. Moreover, callus was induced from young
ovaries when these were incubated on MS medium containing 8.88 μM BA, 10.74 μM NAA, and 9.05 or 18.10 μM 2,4-D. Subsequently,
callus was transferred to MS medium supplemented with kinetin (KT) and NAA for shoot organogenesis. Frequency of shoot regeneration
from petal-derived callus was highest when callus was transferred to medium containing 2.69 μM NAA with either 9.29 or 13.94 μM
KT. Shoot regeneration frequency from ovary-derived callus was highest when this callus was transferred to medium containing
9.29 μM KT and 10.74 μM NAA. Overall, different explant types exhibited different organogenic capacities wherein, young petals
had higher shoot regeneration frequencies than young ovaries. The highest rooting frequency (98.25 ± 3.04%) was obtained when
shoots were transferred to half-strength MS medium without plant growth regulators. Regenerated plantlets were transplanted
to soil mix and acclimatized, yielding a 96.80% survival frequency. Only 0.6% of regenerated plantlets exhibited morphological
changes. The diploid status (2n = 22) of regenerated plantlets was determined using chromosome counts of root-tips. Moreover, inter-simple sequence repeats
were used to assess the genetic fidelity of regenerated plantlets. Overall, regenerated plants shared 90.5–100.0% genetic
similarities with mother plants and 89.0–100.0% similarities with each other. 相似文献
2.
Stem segments of adult plants of Ficus religiosa L. cultured on MS medium containing 1.0 mg/l 2,4-D produced callus. Shoots were regenerated when the induced calli were transferred to medium supplemented with 0.05 to 2.0 mg/l BAP. Callus derived shoots produced roots and developed into plantlets when transferred to medium supplemented with 1.0 mg/l NAA.Abbreviations MS
Murashige and Skoog (1962)
- BAP
6-benzylaminopurine
- NAA
naphthaleneacetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid 相似文献
3.
Plant regeneration was achieved from coleoptile tissue of wheat (Triticum aestivum L. cv. Kharachia-65). Coleoptiles (1.0
- 3.5 cm long) were excised from 2- to 5-d-old seedlings and cultured on Murashige and Skoog's (MS) medium supplemented with
2,4-dichlorophenoxyacetic acid (2,4-D - 0.5, 2.5, and 5.0 mg dm-3). Cream, friable callus was obtained after 6 weeks of inoculation. This callus was sub-cultured on MS medium supplemented
with 2,4-D (2.5 mg dm-3) and 5 % coconut water. After 6 weeks of sub-culturing white, cream or pale, friable, nodular callus was obtained. Plant
regeneration occurred when this callus was sub-cultured on MS medium supplemented with 0.2 mg dm-3 1-naphthalene acetic acid + 1.0 mg dm-3 6-benzylaminopurine. For rooting, regenerated shoots or plantlets were transferred on MS medium supplemented with 0.5 mg
dm-3 indole-3-acetic acid. Rooted plantlets were directly transferred into pots and grown under field conditions. Seed setting
invariably occurred in all plants.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
4.
Mi Jin Jeong Hyun Jin Song Dong Jin Park Ji Yun Min Jin Seong Jo Bo Min Kim Hak Gon Kim Yong Duck Kim Ru Mi Kim Chandrakant S. Karigar Myung Suk Choi 《Plant Cell, Tissue and Organ Culture》2009,98(1):59-65
An efficient plant regeneration protocol for shoot organogenesis from Hovenia dulcis callus cultures was established. Induction of organogenic callus was achieved on Murashige and Skoog (MS) medium supplemented
with 4.65 μM kinetin and 4.5 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Further differentiation of organogenic callus into
primordia, shoot-like structures, and plantlets was achieved on MS medium supplemented with 0.23 μM gibberellic acid (GA3) and 0.46 μM kinetin. Numerous abnormal shoots developed upon transfer of callus to MS medium containing cytokinins, and
these failed to grow further into whole plantlets. However, transfer of ‘abnormal’ shoots to a fresh MS medium lacking cytokinins
resulted in growth of normal shoots. Elongated shoots subsequently were rooted in basal MS medium, and whole plantlets were
established in a soil mix. Analysis of regenerated plants using random amplified polymorphic DNA (RAPD) confirmed the genetic
stability of these regenerant plantlets. 相似文献
5.
Calli were initiated from leaf segments (~0.5 × 0.5 cm) of daylily incubated on Murashige and Skoog (MS) (1962) medium supplemented with 2,4-dichlorophenoxyacetic acid (2.4-D), and either benzyladenine (BA) or thidiazuron (TDZ). The highest frequency of callus induction was observed on medium with 6.79 μM 2,4-D plus either 4.55 or 6.81 μm TDZ. A period of callus maintenance on medium containing 5.37 μM naphthaleneacetic acid (NAA) plus 2.22 or 4.44 μM BA was necessary following induction to improve the quality of the callus, and significantly increase the frequency of embryogenic-like callus formation and shoot regeneration once calli were transferred to light. Over 70% of the regenerated shoots produced roots on ½ strength MS medium lacking plant growth regulators. The regenerated plantlets were successfully transferred into soil and acclimatized in growth rooms. This is the first report showing that leaf segments can be used for daylily regeneration. 相似文献
6.
Hina Khan Iram Siddique Mohammad Anis Pervaiz Rasheed Khan 《Journal of plant biochemistry and biotechnology.》2011,20(1):84-89
An efficient protocol of shoot organogenesis and plant regeneration from internode derived callus has been developed for Capsicum annuum. Optimal callus was developed from internodal segments on Murashige and Skoog (MS) medium supplemented with 10 μM 2,4-dichlorophenoxy acetic acid (2,4-D) and 2.0 μM 6-benzyladenine (BA). Shoot differentiation was achieved from the surface of callus when transferred on shoot induction medium containing BA and thidiazuron (TDZ) alone or in combination. The highest number of de novo adventitious shoots (25.4?±?1.42) and shoot length (4.6?±?0.37 cm) was recorded on MS medium supplemented with 5.0 μM BA and 2.5 μM TDZ. The individual elongated shoots were rooted well on MS medium supplemented with 1.0 μM Indole-3-butyric acid (IBA). The in vitro raised plantlets with properly developed shoot and roots were acclimatized successfully and grew well in the greenhouse. All the regenerated plants appeared normal with respect to morphology and growth characteristics with 85% survival rate. 相似文献
7.
In vitro organogenesis and plant regeneration from unpollinated ovary cultures of Azadirachta indica
A novel method of organogenesis in neem (Azadirachta indica A. Juss.) from unfertilized ovaries is described. The Murashige and Skoog’s (MS) medium with 9 % sucrose, 1 μM 2,4-dichlorophenoxyacetic
acid (2,4-D) and 5 μM 6-benzylaminopurine (BAP) was the best for callus induction from unfertilized ovaries. However, further
proliferation of callus occurred better on MS medium supplemented with 0.5 μM 2,4-D either alone or in combination with 4.5
μM kinetin. Maximum shoot regeneration (78 %) was observed when calli, induced from ovaries of 4 mm size flower buds and proliferating
on MS + 0.5 μM 2,4-D, were subcultured to MS medium containing 5 μM BAP. Histological analysis revealed that 4 mm sized flower
bud corresponds to a 2-nucleate stage of embryo sac. The shoots were then multiplied by forced axillary branching on MS medium
supplemented with 1.0 μM BAP and 250 mg dm−3 casein hydrolysate. The shoots could be rooted on 1/4 strength MS medium supplemented with 0.5 μM indole-3-butyric acid (IBA)
at a frequency of 79 %. Cytological analysis by root tip squash preparations revealed that all the plantlets were diploids.
These plants were subsequently hardened and established in soil with transplantation rate of 81.8 %. 相似文献
8.
High-frequency plant regeneration through callus initiation from mature embryos of maize (<Emphasis Type="Italic">Zea Mays</Emphasis> L.) 总被引:10,自引:0,他引:10
An efficient maize regeneration system was developed using mature embryos. Embryos were removed from surface-sterilized mature seeds and sliced into halves. They were used as explants to initiate callus on induction medium supplemented with 4.0 mg l–1 2,4-dichlorophenoxyacetic acid (2,4-D). The induction frequency of primary calli was over 90% for all inbred lines tested. The primary calli were then transferred onto subculture medium supplemented with 2.0 mg l–1 2,4-D. Following two biweekly subcultures, embryogenic calli were formed. Inclusion of a low concentration (0.2 mg l–1) of 6-benzylaminopurine (BA) in the subculture medium significantly promoted the formation of embryogenic callus. The addition of silver nitrate (10 mg l–1) also supported an increased frequency of embryogenesis. The embryogenic callus readily formed plantlets on regeneration medium supplemented with 0.5 mg l–1 BA. The regenerated plantlets were transferred to half-strength Murashige and Skoog medium supplemented with 0.6 mg l–1 indole-3-butyric acid to develop healthy roots. The regenerated plantlets were successful on transfer to soil and set seed. Using this system, plantlets were regenerated from seven elite maize inbred lines. The frequency of forming green shoots ranged from 19.8% to 32.4%. This efficient regeneration system provides a solid basis for genetic transformation of maize.Abbreviations BA 6-Benzylaminopurine - 2,4-D 2,4-Dichlorophenoxyacetic acid - IBA Indole-3-butyric acid - KT KinetinCommunicated by M.C. Jordan 相似文献
9.
多年生黑麦草成熟胚再生体系的建立及基因枪转化 总被引:4,自引:0,他引:4
目的:建立以多年生黑麦草成熟胚为起始材料的再生体系,用于基因枪转化。方法:多年生黑麦草成熟种子在附加 5mg L 2,4 D的MS培养基上诱导愈伤组织,转至新继代培养基上产生胚性愈伤组织。分化培养基为无激素MS培养基。再生植株在培养基成分减半的无激素MS培养基生根,之后移栽至土壤。基于这一再生体系,用含有水稻几丁质酶基因RC2 4的质粒pARN6和含有草丁膦乙酰转移酶基因Bar的质粒pDB1,通过基因枪轰击胚性愈伤组织。用附加PPT的继代培养基进行转化植株的抗性筛选。结果:共获得 2 4 3株再生植株。通过PCR进行检测,获得1 8株整合有RC2 4基因的植株,1 5株整合有Bar基因的植株,同时转入 2个基因的植株 2株。 相似文献
10.
Suresh Chand Ashok Kumar Sahrawat 《Journal of plant biochemistry and biotechnology.》2001,10(1):43-47
A simple in vitro protocol was established for high frequency plant regeneration via organogenesis and somatic embryogenesis from the callus cultures derived from immature inflorescence segments of indica rice (Oryza sativa L cvs Safari-17 and Kasturi). Embryogenic and nodular calli were initiated on MSB medium supplemented with 2, 4-D and sucrose (3.0%, w/v). Somatic embryogenesis occurred after transfer of embryogenic calli to MSB medium containing 2.25 μM 2,4-D, 2.2 μM BAP, 2.9 μM thiamine HCl and 244.86 μM L-tryptophan. Plantlet/shoot regeneration occurred after transfer of embryogenic calli to MSB medium containing 17.6 μM BAP and 1.12 μM 2,4-D. Partial desiccation (up to 12, 24, 48, 72 and 96 h) of embryogenic calli prior to transfer to regeneration medium stimulated regeneration frequency. Highly significant (P<0.001) difference was observed for regeneration frequency and average number of plantlets/shoots regenerated per callus in partially desiccated calli in comparison to non-dehydrated calli. Regeneration frequency increased from 33.3% to 80% after 24 h of desiccation treatment to callus cultures of cv. Safari-17, and from 46.7% to 93.3% after 48 h of desiccation treatment to callus tissues of cv. Kasturi. Regenerated shoots were rooted on MSB medium supplemented with 4.9 μM IBA. Plants with well-developed roots were transferred to pots where they grew well and attained maturity. 相似文献
11.
草木樨状黄芪高频离体再生体系的建立 总被引:12,自引:0,他引:12
以草木樨状黄芪无菌苗茎切段为材料,在含1-2mg/L2,4-D和0-0.5mg/L6BA的MS培养基上培养获得大量愈伤组织,愈伤组织诱导率在95%以上,愈伤组织在附加0.2mg/LKT,1mg/L6BA,0或0.5mg/LNAA,500mg/LCH 和200mg/L YE的MS培养基上诱导丛生芽,并进而发育成苗。苗的分化频率为100%。分化苗或其茎的切段在不国源植物激素的1/2MS培养基上可出现根的分化,分化频率达90%以上,再生植株经炼苗后移栽成活率达80%以上。 相似文献
12.
Parthasarathi Bhattacharya Satyahari Dey Nilanjana Das Bimal Chandra Bhattacharyya 《Plant cell reports》1990,9(8):439-442
A procedure for rapid multiplication of Chrysanthemum morifolium RAMAT cv. Birbal Sahni using leaf callus and stem (nodal/internodal) callus as well as node and apical shoots has been developed. Murashige and Skoog's medium (1962) supplemented with 2mg/1 2,4-D yielded good green calli from both leaf and stem segments within 2 weeks. About 1 cm × 1 cm callus regenerated 2–3 shoots after 3 weeks on MS solid medium supplemented with 0.1 mg/l IAA and 0.2 mg/l BAP. Each of the regenerated shoots when transferred to the same shooting medium without agar yielded about 150 new shoots, which in turn regenerated roots after another week in MS half strength or modified White's media (Rangaswamy, 1961). It has been estimated that about 1014 plantlets could be produced in a year from one expiant following the proposed protocol.Abbreviations BAP
6-benzylaminopurine
- IAA
indole-3-acetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- MS
Murashige and Skoog's (1962) medium 相似文献
13.
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
14.
Regeneration of Panax ginseng C.A. Meyer by organogenesis and nuclear DNA analysis of regenerants 总被引:1,自引:0,他引:1
Plant regeneration ability of ginseng (t Panax ginseng C.A. Meyer) via organogenesis was studied. Compact callus was induced
from four different types of explants-leaf, petiole, flower stalk, and root of t in vitro-grown plantlets. Petioles were found
to be the best material for callus induction. Calli induced on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxyacetic
acid (4.5 μM) and kinetin (0.46 μM) were conditioned for two weeks on the same medium. These calli differentiated into adventitious
shoots when cultured on 1/2MS basal medium plus kinetin 4.7 μM and silver thiosulphate 10 μM. An addition of GA3 (2.9 μM) and BA (4.4 μM) to MS basal medium, however, induced high frequency t in vitro flowering (86.1%) and multiple shoot
budding which affected the normal complete development of plantlets. Plantlets with a well-developed root system were obtained
six weeks after regenerated shoots had been transplanted to 1/2 MS20 medium containing IBA 1.2 μM. Nuclear DNA content was
measured to check the stability of their ploidy level. Based on DNA flow cytometric analysis, all the regenerants were typically
diploids as the mother plants were, indicating that nuclear DNA content remained stable during cell differentiation.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
15.
Shahina Parveen Anwar Shahzad Mohammad Anis 《Journal of plant biochemistry and biotechnology.》2012,21(2):213-219
In vitro regeneration was achieved through callus culture derived from cotyledon explants of Cassia angustifolia Vahl. on MS (Murashige and Skoog, 1962) medium. Calli were induced from cotyledon explants excised from aseptic 14?days old seedlings on MS medium containing 2,4-D (2,4-dichlorophenoxy acetic acid) and 2,4,5-T (2,4,5-trichlorophenoxy acetic acid) at different concentrations with 3% sucrose and 0.8% agar. Optimal growth of callus was obtained at 5.0???M 2,4-D, which was proved to be the best for shoot regeneration when sub cultured onto MS medium supplemented with cytokinins either alone or in combination with an auxin. Maximum number of shoots (23.2?±?1.4) were produced at 5.0???M 6-benzylaminopurine (BA) and 0.4???M ??-naphthalene acetic acid (NAA). Regenerated shoots produced prominent roots when transferred to half strength MS medium supplemented with 1.0???M indole-3-butyric acid (IBA) and 5.0???M phloroglucinol (PG). Rooted plantlets thus developed were hardened and successfully established in the soil. This protocol yielded an average of 23 plants per cotyledon explant over a period of 4?months. 相似文献
16.
M. Ramakrishnan S. Antony Ceasar V. Duraipandiyan Melvin A. Daniel S. Ignacimuthu 《In vitro cellular & developmental biology. Plant》2013,49(3):285-293
An efficient somatic embryogenesis and regeneration system was developed for the first time in onion using shoot apex explants. These explants were used to initiate callus in Murashige and Skoog (MS) medium supplemented with 4.0 mg l?1 2,4-dichlorophenoxyacetic acid. The induction frequency of primary callus in this medium was 85.3%. The primary calli were then transferred onto medium supplemented with 2.0 mg l?1 2,4-dichlorophenoxyacetic acid. Following two biweekly subcultures, embryogenic callus formed. Inclusion of a low concentration of 6-benzylaminopurine in the subculture medium promoted the formation of embryogenic callus. The addition of 2.0 mg l?1 glycine, 690 mg l?1 proline, and 1.0 g l?1 casein hydrolysate also increased the frequency of callus induction and embryogenic callus formation. The highest frequency of embryogenic callus (86.9%) and greatest number of somatic embryos (26.3 per callus) were obtained by the further addition of 8.0 mg l?1 silver nitrate. Somatic embryos formed plantlets on regeneration medium supplemented with 1.5 mg l?1 6-benzylaminopurine; addition of 2.0 mg l?1 glycine to the regeneration medium promoted a high frequency of regeneration (78.1%) and plantlet formation (28.7 plants per callus). The regenerated plantlets were transferred to half-strength MS medium supplemented with 1.5 mg l?1 indole-3-butyric acid for root development; the maximum frequency of root formation was 87.7% and the average number of roots was 7.6 per shoot. The regenerated plantlets were successfully grown to maturity after hardening in the soil. This is the first report of somatic embryogenesis and regeneration from shoot apex explants of onion. 相似文献
17.
Vinod Singh Gour S. K. Sharma C. J. S. K. Emmanuel Kant Tarun 《Journal of plant biochemistry and biotechnology.》2007,16(2):151-153
The callus mediated regeneration system for Balanites aegyptiaca (L) Del is reported here. Different explants like apical buds, young thorns and cotyledon pieces from mature tree and root segments from in vitro raised seedlings were used for callus induction on MS medium supplemented with 2.23 μM 2,4-Dichlorophenoxyacetic acid. Seven to eight weeks old calli were transferred on hormone free MS medium in order to get regeneration. Shoot morphogenesis was achieved only from cotyledon-derived callus. The shoots so produced rooted well, when cultured on B5 medium supplemented with 9.84 μM Indole-3-butyric acid. Plantlets have been transferred to the field after two-phase hardening and are performing well. 相似文献
18.
Callus induction and plant regeneration from hypocotyl explants of the forage legume Astragalus adsurgens 总被引:1,自引:0,他引:1
An efficient procedure was developed for inducing callus and plant regeneration using hypocotyl segments of Astragalus adsurgens. The combinations and concentrations of different growth regulators were shown to be critical factors for both the frequency
and the type of callus formation as well as for the potential of callus differentiation. Of the four morphologically distinct
types of calli that were induced, a friable, yellow callus, i.e. type I, induced on MS medium supplemented with 9.0 μM 2,4-dichlorophenoxyacetic acid and 2.2 μM N6-benzylaminopurine (BA), and then transferred to MS medium containing 0.5 μM α-naphthaleneacetic acid and 8.9 μM BA, exhibited the maximum frequency of shoot regeneration (75%). After regenerated shoots were transferred onto half-strength
MS medium without growth regulators, they rooted and complete plants were obtained. Plantlet regeneration from callus cultures
required 7–8 weeks.
Received: 26 February 1997 / Revision received: 28 August 1997 / Accepted: 13 September 1997 相似文献
19.
Manickam V.S. Elango Mathavan R. Antonisamy R. 《Plant Cell, Tissue and Organ Culture》2000,62(3):181-185
The development of stem callus mediated plant regeneration system for Withania somnifera is described. Maximum callus proliferation was obtained on Murashige and Skoog medium supplemented with 2.26 μM 2,4-D. Three-week-old,
white, friable callus was used for shoot regeneration. The maximum shoot regeneration (6.2 ± 0.34 shoots/explant) was achieved
in four weeks when callus was cultured on MS medium fortified with 4.44 μM BA and 0.57 μM IAA. Regenerated shoots were excised
and multiplied (8.4 ± 0.43 shoots/explant) on MS medium supplemented with 4.44 μM of BA. Multiple shoots were divided into
single shoots and were rooted (5.1 ± 0.49 rootlets/shoot) on half strength MS medium supplemented with 9.84 μM of IBA. After
a hardening phase of 3 weeks the plantlets were transferred to the field.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
20.
Long-term culture establishment and efficient in vitro regeneration protocol for Sansevieria cylindrica Bojer ex Hook was developed using leaf derived callus and nodule culture. Profuse callus induction on leaf discs was achieved on Murashige
and Skoog (MS) medium supplemented with 10 μM indole-3-butyric acid (IBA), while a high frequency of nodulation was induced
on 2,4-dichlorophenoxyacetic acid (2,4-D) and 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) containing media. Shoot regeneration
ability from cultured tissues occurred at varying degrees on all media. Through callus culture a maximum of 17.6 ± 0.14 shoots
per culture was formed on medium containing 5μM 6-benzyladenine (BA) and 2 μM α-naphthaleneacetic acid (NAA). Among nodule
cultures, the 2,4-D generated nodules were more proliferative and regenerative as compared to 2,4,5-T induced nodules and
a maximum of 25 ± 0.16 shoots per culture was produced on a medium containing 5 μM BA plus 1 μM NAA. The regenerated shoots
were successfully rooted on a semi-solid half strength MS medium containing 5 μM IBA with an average root number 3.5 ± 0.18
and root length 6.5 ± 0.14 cm. The regenerative ability of callus tissues was steady upto one year, while the nodules retained
the totipotency to regenerate on optimal medium even after 3 years of subculturing. The histological sections of nodules confirm
the typical anatomy exhibiting the vascular elements in bundles with well demarcated cortex and epidermal covering. 相似文献