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1.
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
2.
In vitro induction of multiple shoots and plant regeneration in cotton (Gossypium hirsutum L.) 总被引:5,自引:0,他引:5
D. C. Agrawal A. K. Banerjee R. R. Kolala A. B. Dhage A. V. Kulkarni S. M. Nalawade S. Hazra K. V. Krishnamurthy 《Plant cell reports》1997,16(9):647-652
Induction of multiple shoots in cotton (Gossypium hirsutum L. cv. Anjali-LRK 516) has been achieved with cotyledonary nodes devoid of cotyledons and apical meristems. Explants from 35-day-old seedlings yielded the maximum number of shoots (4.7 shoots/explant) using Murashige and Skoog (MS) basal medium supplemented with 6-benzylaminopurine and kinetin (2.5 mg/1 each). Explants from 35-day-old seedlings raised in glass bottles produced a higher number of multiple shoots (8.3 shoots/explant) than those grown in glass tubes and cultured on the same shoot induction medium. Elongation of multiple shoots was obtained on liquid or agar MS basal medium without phytohormones. In vitro shoots were rooted on half-strength agar-solidified MS basal medium or with 0.05 or 0.1 mg/1 naphthaleneacetic acid. Hardening and survival of tissue culture plantlets was 95% under greenhouse conditions.Abbreviations
BAP
6-Benzylaminopurine
- GA3
Gibberellic acid
- MS
Murashige and Skoog medium
-
NAA
-Napthaleneacetic acid 相似文献
3.
Root segments obtained from aseptically germinated seedlings of Brassica napus cv. Westar were used to optimize conditions for high-frequency shoot bud differentiation. The presence of low kinetin (0.5 M) and relatively high indole-butyric acid (1.0 M) levels facilitated optimum shoot bud differentiation. Modified MS medium (MMS) was superior to the other three basal media tested (MS, B5 and White's). Elevated sodium dihydrogen phosphate levels increased the differentiation of shoot buds. Increasing or decreasing the level of sucrose from 3% reduced the frequency of explants forming shoot buds. Addition of glutamine enhanced both the frequency of responding explants, as well as the number of shoots per responding explant. Root segments from 13-day-old seedlings produced the highest response (58%) in the presence of 100 mg l-1 glutamine. The position of the segment on the main root, size, and the presence or absence of lateral roots altered the morphogenic response. Sealing of the donor seedling cultures with Parafilm® instead of Stretch' n seal® resulted in a higher production of shoot buds, although root segment cultures were not affected by the type of sealing. Spontaneous rooting occurred on all developed shoots.Dedicated to Dr. Friedrch Constabel on the occasion of his 60th birthday 相似文献
4.
Viable protoplasts of Vigna sublobata L. were isolated enzymatically from hypocotyls of axenic seedlings. Protoplast yields were dependent upon seedling age, with maximum yields (2.25 ± 0.35 × 106 g fwt–1) from seedlings aged 6 d. Protoplasts regenerated cell walls and underwent sustained divisions when cultured in either agarose-solidified or liquid K8P medium. The plating density affected the division frequency and plating efficiency; the division frequency (68 ±0 6.0%) was maximum at 4.0 × 104 ml–1 while plating efficiency was maximum (1.3 ± 0.1%) at 5.0 × 104 ml–1. Dividing protoplasts developed into microcalli, which produced glossy green compact nodular calli on transfer to 8.0 gl–1 w/v agar-solidified medium containing MS salts, B5 organic components, 30 g l–1 sucrose, NAA (0.2–0.5 mg l–1), zeatin riboside (0.5–2.0 mg l–1) and GA3 (0.5–1.0 mg l–1). These calli, after sub-culture on the same medium, produced shoot buds which underwent elongation following transfer of tissues to 6.0 g l–1 agar-solidified B5 medium containing 30g l–1 sucrose, IBA (0.01 mg l–1) and BAP (1.0 mg l–1). Elongated shoots developed roots after transfer to 8.0g l–1 agar-solidified, hormone-free MS medium with 30 g l–1 sucrose.Abbreviations IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- BAP
6-benzyladenine or benzylaminopurine
- B5
medium after Gamborg et al (1968)
- 2,4-D
2,4-dichlorophenoxyacetic acid
- GA3
gibberellic acid
- 2,i-P
6-(--dimethylallylamino) purine
- MS
medium after Murashige and Skoog (1962)
- NAA
1-naphthaleneacetic acid 相似文献
5.
Lance C. Heath Shih-Foong Chin Donald Spencer Thomas J. V. Higgins 《Plant Cell, Tissue and Organ Culture》1993,35(1):43-48
Regeneration of subterranean clover (Trifolium subterraneum L.) was achieved by both shoot organogenesis and somatic embryogenesis. Shoots derived via organogenesis were initiated from the hypocotyls of mature imbibed seed. The hypocotyl, including the emerging radicle, was sliced longitudinally into two halves and cultured on shoot induction medium. After 30 days, adventitious shoots were formed from the hypocotyl region while the radicle showed no development. Shoots were then subcultured onto shoot multiplication medium and finally onto a root initiation medium. Histological studies revealed that shoots arose de novo and did not originate from pre-existing meristems. In the second regeneration protocol, shoot apical meristems from young seedlings were induced to form callus. Following four to six weeks culture in the dark, somatic embryos appeared spontaneously on the calli. A majority of embryos had a well-defined root pole, two cotyledonary lobes, and were capable of germination, albeit at a low frequency. Regenerated plants obtained from both protocols appeared phenotypically normal. 相似文献
6.
紫茉莉(Mirabilis jalapa)观赏价值较高,是一种重要的污染修复植物.组织培养技术为植物品种改良和选育的重要途径,但紫茉莉离体快繁方面的研究尚未见有相关报道.该研究以紫茉莉叶片和茎段为外植体,通过观察和统计外植体愈伤组织和不定芽的诱导情况,分析不同植物生长物质对紫茉莉植株再生的影响.结果表明:紫茉莉带芽茎段比较适合丛生芽的诱导,当带芽茎段在MS+1.0 mg·L-16-BA+1.5 mg·L-1 KT+1.0 mg·L-1 NAA+0.05 mg·L-1 TDZ培养基中培养时,不定芽的增殖系数较高.无论是MS或1/2MS培养基,都可诱导不定根的产生,其中生根效果较好的培养基为1/2 MS+0.5 mg·L-1 NAA.该研究结果探索了紫茉莉组织培养的最适条件,根据愈伤组织诱导率和不定芽的增殖系数筛选出了适宜不定芽诱导的培养基类型,根据不定芽生根情况确定了最佳的生根诱导培养基,为建立紫茉莉高效稳定的再生和遗传转化体系奠定了基础. 相似文献
7.
Sriskandarajah Sridevy Frello Stefan Serek Margrethe 《Plant Cell, Tissue and Organ Culture》2001,67(3):295-298
Efficient protocols have been developed to induce adventitious shoots in different types of explants of Campanula carpatica Jacq. More than five shoots per explant developed on hypocotyls of 5-week-old seedlings after 2 weeks of culture. Hypocotyls produced twice as many shoots as the cotyledons. TDZ proved to be about 6 times more efficient than BA. NAA had to be added to the regeneration medium to obtain the optimal balance of auxin and cytokinin to induce shoot regeneration. Significant differences were noted between different growth regulator concentrations in their effects on shoot organogenesis. BA induced double the number of callus clumps as TDZ. Incubation of explants in the dark produced about 6 shoots per explant while those in the light produced about 2 shoots per explant. Explants derived from 5-week-old seedlings were five times more regenerative compared to those derived from 15-week-old seedlings. Explants from cv. White Uniform were more organogenic than those from cv. Blue Clip. Root segments were also found to form shoots when treated with CPPU. 相似文献
8.
Summary An efficient and reproduciblein vitro culture system has been developed for regeneration of multiple shoot clumps from intact seedlings of both lowland and upland
cultivars of switchgrass (Panicum virgatum L.). The multiple shoots were induced on Murashige and Skoog medium supplemented with various combinations of 2,4-dichlorophenoxyacetic
acid (2,4-D) and 1-phenyl-3-(1,2,3-thiadiazol-5YL)-urea (thidiazuron or TDZ). Maximum response was obtained with 4.5 μM 2,4-D and 18.2 μM TDZ. These shoots proliferated and rooted efficiently on MS medium without growth regulators. The developmental pattern of
the multiple shoots indicated their origin from the enlarged shoot apex via proliferation of axillary buds and subsequent
reprogramming of shoot meristems followed by secondary differentiation of adventitious shoots The simplicity of the protocol
and direct production of multiple shoots make this a potential system that is highly attractive and amenable for microprojectile-mediated
gene transfer. 相似文献
9.
Axillary shoot induction and plant regeneration were obtained in Plantago ovata. The optimum medium for inducing axillary shoots was Murashige & Skoog (MS) medium [5] supplemented with 4.6 M kinetin and 0.05 M NAA. Rooting of shoots was best on half-strength MS medium containing 5.0 M IBA and 0.05 M kinetin. The regenerated plants were similar to the control plants in karyotypic and phenotypic details. 相似文献
10.
Thao Nguyen Thi Phuong Ozaki Yukio Okubo Hiroshi 《Plant Cell, Tissue and Organ Culture》2003,72(3):285-289
Callus induction from petiole explants has been achieved in Alocasia micholitziana `Green Velvet'. The highest percentage (71%) of explants inducing callus was obtained on MS medium supplemented with 0.5 M 2,4-D and 0.5 M kinetin in the dark after 4 months of culture. Shoots were regenerated at the highest frequency of 33.3% under light condition when 0.5 M BA was added to MS medium with the average of 7.8±2.3 shoots per callus explant. The callus-derived shoots rooted on hormone free MS medium and within 4 weeks the plantlets were ready for acclimatization. The regenerated plants appeared morphologically similar to mother plants. 相似文献
11.
Monacelli Barbara Pasqua Gabriella Cuteri Angelina Vitali Alberto 《Plant Cell, Tissue and Organ Culture》1999,58(2):81-85
A protocol for in vitro plant regeneration through organogenesis was established for Vismia guianensis(Hypericaceae), a species that produces an anti-cancer compound. The highest mean number of shoots per gram of callus (57.33)
was obtained on Murashige and Skoog medium supplemented with 4.44 μM 6-benzyl-aminopurine, 5.70 μM indole-3-acetic acid and
12.88 μM gibberellic acid. Rooting was favoured by the addition of 10 μM indole-3-butyric acid, and by sucrose concentrations
higher than 1%.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
12.
Direct shoot organogenesis and plant regeneration in safflower 总被引:1,自引:0,他引:1
Summary Adventitious shoot buds were induced directly on the adaxial surface of the cotyledons of eight safflower cultivars after
14 d of culture initiation on Murashige and Skoog's (MS) medium supplemented with various levels of 6-benzylaminopurine (BA).
Maximum shoot organogenesis of 54.4% with 10.2 shoots per responding cotyledon was obtained with 8.87 μM BA in the cv. S-144. Regenerated shoots were classified into three groups on the basis of their morphological features and
were found to be correlated with the levels of BA. The highest number of normal shoots was obtained from 2.2 μM BA treatment. The regenerated shoots of group I (normal shoots) were rooted on half-strength MS medium supplemented with
5.3 μM α-naphthaleneacetic acid, 3% sucrose and 0.8% bacto-agar. Rooted plantlets were successfully transferred to soil and appeared
morphologically normal. Histological studies revealed that shoot buds originated adventitiously from subepidermal cells. 相似文献
13.
In vitro proliferation of shoots and regeneration of cotton 总被引:3,自引:0,他引:3
Gupta Shiv K. Srivastava Alok K. Singh Pradhyumna K. Tuli Rakesh 《Plant Cell, Tissue and Organ Culture》1997,51(2):149-152
Shoot proliferation from different explants of several Indian cultivars of cotton was studied in culture. Cotyledonary nodes
taken along with the shoot apex of seedlings produced multiple shoots on modified MS nutrient agar supplemented with cytokinins.
6-Benzyladenine was most effective in inducing growth of multiple shoots. Explants of several genotypes formed organogenic
masses that differentiated to secondary shoots on repeated subculture. The isolated shoots were rooted on basal medium supplemented
with naphthaleneacetic acid and were transferred to soil after acclimatization.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
14.
P. Sansberro H. Rey L. Mroginski M. Collavino 《In vitro cellular & developmental biology. Plant》1999,35(5):401-402
Summary Nodal segments as well as shoot tips and apical meristems of 2-yr-old “maté” plants (Ilex paraguariensis St. Hil.) were cultured in vitro to establish micropropagation systems. Maximum shoot regeneration was achieved when nodal
segments were cultured with 1/4 Murashige and Skoog (MS) medium with 3% sucrose. We induced roots to differentiate by transferring
the regenerated shoots onto the same medium, solidified with 2.5 g Phytagel per 1 and supplemented with indole-3-butyric acid
(7.4 μM) and finally transferring shoots to 1/4 MS medium with 3% sucrose and lacking growth regulators. Plants were successfully
established in soil. 相似文献
15.
In vitro organogenesis was achieved from callus derived from hypocotyl explants of Cucumis sativus L. cv. Poinsett 76. Calli were induced from hypocotyl explants excised from 7-d-old seedlings grown on Murashige and Skoog
(MS) medium containing 87.64 μM sucrose, 0.8 % agar, 3.62 μM 2,4-dichlorophenoxy acetic acid and 2.22 μM 6-benzyladenine (BA).
Regeneration of adventitious buds from callus (25 shoots explant−1) was achieved on MS medium supplemented with 8.88 μM BA, 2.5 μM zeatin and 10 % coconut water after two subcultures in the
same medium at 30-d interval. Gibberellic acid (1.75 μM) favoured shoot elongation and indole 3-butyric acid (7.36 μM) induced
rooting. Rooted plants were hardened and successfully established in soil. 相似文献
16.
Charlene Chang Ben A. Moll Kathleen B. Evenson Mark J. Guiltinan 《Plant Cell, Tissue and Organ Culture》1996,45(1):61-66
In vitro plantlet regeneration systems for the seed geranium (Pelargonium x hortorum Bailey) using cotyledon, hypocotyl and root explants were optimized by studying the influence of seedling age, growth regulators and excision orientation on organogenesis. Indole-3-acetic acid combined with zeatin yielded the highest rate of shoot production on cotyledon explants (0.2–2 shoots per explant). More shoots were produced on explants cut from the most basal region of cotyledons from 2 to 4-day-old seedlings than from older seedlings or more distal cut sites. Hypocotyl explants produced the highest number of shoots, up to 40 shoots per explant, on indole-3-acetic acid (2.8–5.6 mM) + zeatin (4.6 mM) or thidiazuron (4.5 mM). Maximum shoot formation (0.3–1.4 shoots per explant) on root explants occurred when they were cultured on medium containing zeatin. Regenerated shoots rooted best on a basal medium containing no growth regulators. There were substantial differences among cultivars in shoot formation from each of the explant systems.Abbreviations BA
6-benzylaminopurine
- 2,4-d
2,4-dichlorophenoxyacetic acid
- IAA
indole-3-acetic acid
- NAA
naphthaleneacetic acid
- TDZ
thidiazuron 相似文献
17.
An effective protocol has been developed for plant regeneration from cotyledon explants of Swainsona salsula Taubert (Saline swainsona), a medicinal and agronomic shrub. Adventitious shoots were obtained from 83.2% of cotyledon explants
from 3-day seedlings cultured on Murashige and Skoog (MS) medium containing 2.0 mg l−1 thidiazuron (TDZ), with an average of 9.3 shoots per explant. Individual elongated shoots were rooted on half strength MS
medium supplemented with 2.0 mg l−1 indole-3-butyric acid (IBA), with 59.3% success. Regenerated plants with well developed shoots and roots were successfully
transferred to soil, without detectable variants. Histological observation revealed that shoots developed from cotyledon explants
via organogenesis, with little callus.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
18.
G.-Y. Kai L.-M. Dai X.-Y. Mei J.-G. Zheng W. Wang Y. Lu Z.-Y. Qian G.-Y. Zhou 《Biologia Plantarum》2008,52(3):557-560
An efficient in vitro plant regeneration system from leaves of Ophiorrhiza japonica Blume was established for the first time. Callus formation rate was more than 90.4 % from leaf segments on Murashige and
Skoog (MS) supplemented with either α-naphthaleneacetic acid (NAA) alone or in combination with 6-benzyladenine (BA). The
highest shoot regeneration (78.9 %) was achieved on MS medium containing 2.0 mg dm−3 BA and 0.2 mg dm−3 NAA, with an average of 9.4 shoots developed per leaf segment. Shoot regeneration was also improved when the leaf explants
were cultured in MS basal medium supplemented with 0.5 % (m/v) polyvinylpyrrolidone (PVP). The leaf explants from seedlings
with age of about 18–27 d showed the highest shoot regeneration. The regenerated shoots were rooted on half-strength basal
MS medium supplemented with 0.5 mg dm−3 indole-3-butyric acid (IBA), which averagely produced 24.8 roots per shoot. The plantlets were transferred to soil, where
100 % survived after 1 month of acclimatization. 相似文献
19.
Jameel M. Al-Khayri Feng H. Huang Teddy E. Morelock Tahani A. Busharar 《In vitro cellular & developmental biology. Plant》1992,28(2):64-66
Summary A system for the regeneration of spinach (Spinacia oleracea L.) from mature dry seed explants has been established. The response of two commercial spinach cultivars, ‘Grandstand’ and
‘Baker’, was examined. Callus proliferation was most prominent on MS medium supplemented with 9.3 μM of 6-furfurylaminopurine (kinetin) and 3.39 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Adventitious shoot formation was observed within 8 wk after callus was transferred
onto regeneration medium. Shoot regeneration was best from callus induced on 9.3 μM kinetin and 4.56 μM 2,4-D. The regeneration medium contained 9.3 μM kinetin, 0.045 μM 2,4-D, and 2.89 μM gibberellic acid (GA3). Shoots were rooted on hormone-free medium, and plants grown in a greenhouse showed normal phenotype. This system is beneficial
in rapid propagation of spinach plants, particularly when only a limited number of seeds are available. 相似文献
20.
In vitro regeneration and cytological characterization of shoots from leaf explants of three accessions of Solanum commersonii 总被引:2,自引:0,他引:2
Teodoro Cardi Vittoria Iannamico Fabrizio D'Ambrosio Edgardo Filippone Paul F. Lurquin 《Plant Cell, Tissue and Organ Culture》1993,34(1):107-114
In vitro culture of explants were used to apply genetic or cell engineering techniques to the sexually incompatible potato relative Solanum commersonii (2n=2x=24) Three accessions of S. commersonii were tested for regeneration from leaf explants using six different protocols. A two step-regeneration procedure gave the best results. Genetic variability for regeneration ability was found between accessions, and between clones within accessions. The accession PI 472834 regenerated at highest frequency. Clones with high regeneration ability were selected. Approximately 60% of regenerated plants were diploids and 40% were tetraploids. A very low frequency of chimeras was found. Leaf shape and chloroplast counts in guard cells were shown to be quick and reliable methods for estimating ploidy levels. Use of the diploid and tetraploid regenerants obtained for potato breeding is discussed.Abbreviations BAP
6-benzylaminopurine
- EBN
Endosperm Balance Number
- GA3
gibberellic acid
- IAA
indole-3-acetic acid
- MS
Murashige and Skoog
- NAA
1-naphthalene-acetic acid
- ZEA
zeatin 相似文献