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1.
Slavica Ninković Tatjana Djordjević Branka Vinterhalter Branka Uzelac Aleksandar Cingel Jelena Savić Svetlana Radović 《Plant Cell, Tissue and Organ Culture》2010,103(1):81-91
Agrobacterium rhizogenes A4M70GUS-mediated transformation of two local breeding lines of sugar beet was obtained using 4-week-old seedlings. Root
formation efficiency was 61.54% for SBa genotype and 36.36% for SBb genotype. Five highly proliferated hairy root lines have
been established in liquid hormone-free MS medium. Transgenic nature of the hairy root clones was evaluated by GUS assay,
PCR and RT-PCR analyses. Hairy root-derived calli were induced using different plant growth regulators (PGRs): auxin, auxin/cytokinin
and cytokinin. The best callus induction response was achieved on MS medium containing both 1 mg/l 2,4-dichlorophenoxyacetic
acid (2,4-D) and 1 mg/l thidiazuron (TDZ). Globular embryo-like structures were observed in friable callus after its prolonged
cultivation on MS medium supplemented with TDZ and giberellic acid (GA3) at 1 mg/l each, followed by growth on MS medium containing 1% glucose and 0.5 mg/l 2,3,5-triiodobenzoic acid (TIBA). Histological
analysis revealed somatic embryos at different stages of development in hairy root-derived callus of sugar beet. 相似文献
2.
Li Li Jing Wang Wei Wang Yang Lu Yuliang Wang Genyu Zhou Guoyin Kai 《Biotechnology and Bioprocess Engineering》2008,13(5):606-612
In this study, an efficient transformation system for the medicinal plant Anisodus acutangulus was successfully developed and optimized using Agrobacterium rhizogenes. Three bacterial strains, A4, R1601, and modified C58C1 and three explant types, leaf blade, petiole, and stem, were examined.
The highest transformation efficiency of 94.44% was achieved using strain C58C1 with stem explants. Over 20 independent hairy
root lines were successfully established with strain C58C1 using stem explants, all of which contained the ro/B and ro/C genes as confirmed by polymerase chain reaction (PCR). Out of four media compositions, the liquid 1/2 MS medium was found
the most suitable for hairy root growth. The maximum biomass of one hairy root line increased up to 80 times in liquid 1/2
MS medium after a 30 day culture period. Different hairy root lines displayed a varied capacity for tropane alkaloid production
and the best hairy root line (T4) from the C58C1-stem combination produced up to 10.21 mg/g (dw) of hyoscyamine, which was
about 1.5-fold higher than in the wild type plants. To our knowledge, this is the first report to demonstrate the production
of tropane alkaloids in hairy roots of A. acutangulus. 相似文献
3.
A rapid and efficient in vitro plant regeneration method was developed for Matteuccia struthiopteris (L.) Todaro (Ostrich fern). Side shoots, originating in meristems of sectioned rhizomes, were used as explant material. A
very high rate of meristem multiplication was achieved by culturing the explants in half-strength MS liquid medium supplemented
with 2.0 mg/l N-(4-Pyridyl)-N′-phenylurea (4-PU) and 0.5 mg/l thidiazuron (TDZ). Multiplication of the shoot primordia was
faster in suspension culture than on solid medium. Rhizogenesis and growth of regenerants were best achieved on hormone-free
one-quarter-strength MS solid medium amended with 0.4% agar and 1.0% activated charcoal. Regenerated plantlets continued to
grow after transfer to soil in a phytotron.
Received: 19 March 1998 / Revision received: 17 July 1998 / Accepted: 3 August 1998 相似文献
4.
《Plant science》1986,47(1):35-43
Plants were regenerated from cotyledonary and root explants of cucumber (Cucumis sativus L.) cultivars and breeding lines of diverse sex type, growth habit, and processing quality and from cotyledonary explants of muskmelon (C. melo L.). Somatic embryogenesis was induced on a medium consisting of Murashige and Skoog (MS) salts supplemented with 1.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D), 1.0 mg/l α-naphthaleneacetic acid and 0.5 mg/l 6-benzylaminopurine. Embryos matured on the same medium without 2,4-D, and developed into normal plants on a hormone-free MS medium. Cucumber plants were also regenerated from cotyledonary protoplasts using a modified tomato protocol. 相似文献
5.
Songul Gurel Mehmet Cengiz Baloglu Ekrem Gurel Huseyin Avni Oktem Meral Yucel 《Plant Cell, Tissue and Organ Culture》2011,106(2):261-268
The effects of a two-stage pretreatment of seedlings on the subsequent shoot regeneration capacity were investigated. Pretreated
seedlings were obtained by germinating seeds on three different germination media and then further culturing on six different growth media. Lamina and petiole explants of two sugar beet (Beta
vulgaris L.) breeding lines were then excised from the pretreated seedlings and cultured on five different shoot regeneration media. In both breeding lines, petiole explants produced significantly more shoots than lamina explants with higher frequencies
of organogenic capacities; petiole explants of the lines M1195 and ELK345 produced a mean of 2.1 and 2.7 shoots per explant
while their lamina explants produced 1.5 and 2.2 shoots per explant, respectively. A genotypic variation was evident as the
line ELK345 was more productive for shoot development from both types of explants. In overall comparisons of different germination, growth and regeneration media, germination medium was most effective when supplemented with 0.5 mg/l 6-benzyladenine (BA) while both growth and regeneration
media were most productive when contained a combination of 0.25 mg/l BA and 0.10 mg/l indole-3-butyric acid (IBA). Of all
the treatments tested, the highest mean number of shoots per explant (8.3 shoots) and frequency of organogenic explants (75.6%)
were obtained on regeneration medium supplemented with 0.25 mg/l BA and 0.10 mg/l IBA when petiole explants of the line ELK345
were excised from the seedlings that had been germinated on medium containing 0.5 mg/l BA followed by further growth on medium
containing 0.25 mg/l BA and 0.10 mg/l IBA. 相似文献
6.
Philip John Whitney 《Plant Cell, Tissue and Organ Culture》1996,46(2):109-115
This work describes the growth of rye root organ cultures which were capable of being repeatedly subcultured in hormone-free medium. They showed morphological characteristics, growth rate, inability to produce shoots, and response to auxins and cytokinins similar to those of the Agrobacterium rhizogenes (Ri plasmid) transformed hairy root cultures of tobacco and red beet which were used for comparison. The root cultures of rye were initiated from callus produced on a medium containing the growth regulators (plant hormones) 2,4-d and kinetin, then transferred to hormone-free medium. However not all rye explants gave rise to callus that would differentiate into stable hairy root cultures and rye seedling root explants did not grow if placed directly on a hormone-free medium. Rice and wheat produced callus and roots on a medium containing hormones but root organ cultures could not be maintained on a hormone-free medium. 相似文献
7.
Boon Chin Tan Chiew Foan Chin Peter Alderson 《Plant Cell, Tissue and Organ Culture》2011,105(3):457-463
An indirect in vitro plant regeneration protocol for Vanilla planifolia has been established. Juvenile leaf and nodal segments from V. planifolia were used as explants to initiate callus. Nodal explants showed better callus initiation than juvenile leaf explants, with
35.0% of explants forming callus when cultured on Murashige and Skoog (MS) basal medium supplemented with 2.0 mg/l 1-naphthylacetic
acid (NAA) and 1.0 mg/l 6-benzyladenine (BA). Almost 10.0% of juvenile leaf explants were induced to form callus on the MS
basal medium containing 2.0 mg/l NAA and 2.0 mg/l BA, whereas no callus formed in the presence of any concentrations of 2,4-dichlorophenoxyacetic
acid (2,4-D) and BA. After 8 weeks, callus generated was transferred to MS basal medium containing 1.0 mg/l BA and 0.5 mg/l
NAA. A mean number of 4.2 shoots per callus was produced on this medium, with a mean length of 3.8 cm after 8 weeks of culture.
Roots formed on 88.3% of plantlets when they were cultured on MS medium supplemented with 1.0 mg/l NAA, with a mean length
of 4.4 cm after 4 weeks of culture. Of the rooted plantlets, 90.0% survived acclimatisation and were making new growth after
4 weeks. 相似文献
8.
Abstract A high-efficiency plant regeneration protocol based on somatic embryo formation for Huining Roquette, an interesting ecotype of Eruca sativa Mill, was established for future transgenic applications. On Murashige and Skoog (MS) medium containing 2,4-dichlorophenoxyacetic acid (2,4-D), alone or in combination with 6-benzylaminopurine (BA) or kinetin (KT), the cotyledon explants, cotyledon petioles, and hypocotyls all produced embryogenic callus (ECs) or somatic embryos (SEs) to different extents. After transferring onto hormone-free MS medium, the ECs or SEs from the different explants and media, all of them developed shoots with a frequency of 6–48%, and then produced roots with a frequency of 2–29%. As regards the probability of shoot differentiation, cotyledon explants appeared similar to hypocotyls, but superior to cotyledon petioles; 2,4-D + KT worked more effectively than 2,4-D alone and 2,4-D + BA for callus induction and shoot differentiation. The optimal hormone combinations for plant regeneration of cotyledon, cotyledon petiole, and hypocotyl explants were 1.0 mg/l 2,4-D + 0.1 mg/l KT, 0.8 mg/l 2,4-D + 0.3 mg/l BA, and 1.0 mg/l 2,4-D + 0.3 mg/l KT, respectively. MS medium with 60–80 g/l sucrose was the most effective for improving SE maturation and germination. 相似文献
9.
Muthiah Joe Virgin Largia Lakkakula Satish Rajaiah Johnsi Jayabalan Shilpha Manikandan Ramesh 《World journal of microbiology & biotechnology》2016,32(8):131
Agrobacterium rhizogenes mediated transformation has been experimented in leaf explants of the memory herb Bacopa monnieri in order to assess the regeneration potential of hairy roots (HR) followed by the elicitation of transformed plants for increased Bacoside A production. Out of the four strains tested, A4 and MTCC 532 derived HR exhibited regrowth in MS basal medium while MTCC 2364 derived HR showed regeneration in MS medium supplemented with suitable phyto hormones. R1000 derived HR possessed no regeneration potential. Comparable to A4, MTCC 532 derived HR displayed maximum regrowth frequency of about 85.71 ± 1.84 % with an increase in biomass to threefold. Therefore, five HR plant lines (MTCC 532 derived) were generated and maintained in MS basal liquid medium in which HR3 topped the others in producing a huge biomass of about 67.09 ± 0.66 g FW. PCR amplification and southern hybridization analysis of rol A gene (280 bp) has been performed in order to confirm the transformation process. Moreover, HR3 plant line has accumulated highest total phenolic content of about 165.68 ± 0.82 mg GAE/g DW and highest total flavonoid content of about 497.78 ± 0.57 mg QRE/g DW when compared to other lines and untransformed controls. In addition, HR3 plant extract showed 85.58 ± 0.14 % of DPPH (2, 2-diphenyl-1-picryl hydrazyl) inhibition displaying its reliable anti oxidant potential. Further on elicitation with 10 mg/L chitosan for 2 weeks, HR3 has produced 5.83 % of Bacoside A which is fivefold and threefold increased production when compared to untransformed and transformed unelicited controls respectively. This is the first report on eliciting HR plants for increased metabolite accumulation in B. monnieri. 相似文献
10.
Summary A simple and effective procedure has been developed for plantlet regeneration from cotyledon-derived callus of the medicinally
important herb and ornamental species, Incarvillea sinensis. An average of 18.4 adventitious shoots per explant were obtained from 100% cotyledon explants cultured on half-strength
Murashige and Skoog (MS) medium containing 1.0 mg l−1 6-benzylaminopurine for 3 wk, followed by another 4 wk on hormone-free 1/2×MS medium. The cotyledon explants continued to
expand and regenerate new shoots upon repeated subculturing onto fresh medium. Most regenerated shoots (66.9%) were rooted
on 1/4×MS mediumcontaining 1.0 mg l−1 indole-3-acetic acid, with an average of about 3.8 roots per shoot. Regenerated plants with well developed shoots and roots
were successfully acclimatized in soil and were normal phenotypically. 相似文献
11.
High Frequency Somatic Embryogenesis in Cotton 总被引:3,自引:1,他引:2
Y. Aydin Z. Ipekci T. Talas-Oğraş A. Zehir K. Bajrovic N. Gozukirmizi 《Biologia Plantarum》2004,48(4):491-495
A highly reproducible system for efficient somatic embryogenesis was developed to regenerate plantlets from cotton (Gossypium hirsutum L.) cultivars (Nazilli M-503 and Nazilli 143). Shoot apices, hypocotyls and nodes of 10-d-old seedlings were used as explants. High frequency (100 %) embryogenic calli was initiated from all tested explants on Murashige and Skoog (1962) (MS) media supplemented with 1 g dm–3 polyvinylpyrrolidone (PVP), 1 mg dm–3 6-benzylaminopurine (BAP), 0.5 mg dm–3 kinetin for Nazilli M-503 and 1 g dm–3 PVP, 2 mg dm–3 BAP, 0.5 mg dm–3 kinetin for Nazilli-143. Globular stage somatic embryos were produced 4 months after transfer to hormone-free MS medium supplemented with 1 g dm–3 PVP. Subsequent subculture of globular embryos every 3 weeks on hormone-free MS medium led to the development of torpedo and cotyledonary stage embryos with the frequency of 75 and 83.2 % from hypocotyl explants of Nazilli M-503 and Nazilli-143, respectively. Afterwards, mature somatic embryos were isolated and cultured on hormone-free MS medium for germination and development into plantlets. The highest germination frequency (42.9 %) for Nazilli M-503 somatic embryos were obtained on hormone-free MS medium after 5 months with hypocotyl explants, whereas germination frequencies of Nazilli-143 embryos from hypocotyl, node and apex explants varied between 22 – 30 %. 相似文献
12.
Transformation of <Emphasis Type="Italic">Saussurea medusa</Emphasis> for hairy roots and jaceosidin production 总被引:2,自引:0,他引:2
Axenically grown Saussurea medusa plantlets were inoculated with four Agrobacterium rhizogenes strains, and hairy root lines were established with A. rhizogenes strain R1601 in N6 medium. PCR and Southern hybridization confirmed integration of the T-DNA fragment of the Ri plasmid from A. rhizogenes into the genome of S. medusa hairy roots. In N6 medium, maximum biomass of the hairy root cultures was achieved [8 g (dry weight) per liter; growth ratio 35-fold] after 21 days of culture. The amount of jaceosidin extracted from the hairy root cultures was 46 mg/l (production ratio of 37-fold) after 27 days of culture. The maximum jaceosidin content obtained using N6 medium was higher than that obtained with Modified White, MS or B5 medium. In N6 medium, the tip segments were more efficient for hairy root growth and jaceosidin production than the middle and basal regions of the root.Abbreviations AS Acetosyringone - BA Benzyladenine - cef Cefotaxime sodium - DW Dry weight - FW Fresh weight - HPLC High-performance liquid chromatography - IAA Indole-3-acetic acid - km Kanamycin - NAA -Naphthaleneacetic acid - SDS Sodium dodecyl sulfate 相似文献
13.
Hairy roots of Plumbago indica were established at high frequency (90 %) by infecting leaf explants with Agrobacterium rhizogenes strain ATCC 15834. The axenic root cultures were established under darkness in hormone-free liquid Murashige and Skoog medium
containing 3 % sucrose. The highest plumbagin content was found to accumulate in roots at their exponential phase of growth.
A low pH (4.6) and a low concentration of sucrose (1 %) were beneficial for root growth in darkness, while pH 5.6 and 3 %
sucrose under continuous irradiance enhanced plumbagin accumulation in roots up to 7.8 mg g−1(d.m.). Direct shoot regeneration from hairy root culture was also achieved under continuous irradiance, thus indicated an
easy way of obtaining transformed P. indica plants. 相似文献
14.
15.
A three-stage procedure for embryogenesis in Trachyspermum ammi was developed from cotyledon and cotyledonary node explants cultured in Murashige and Skoog (MS) liquid medium supplemented
with 0.2 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D). Globular somatic embryos without intervening callus phase developed in 4 wk. The
development of embryos to heart and torpedo stages required second-stage subculture of the explants (along with developing
embryos) in liquid medium with lower concentrations of 2,4-D. Further development of embryos required a third-stage subculture
in hormone-free liquid medium supplemented with 100 mg l−1 casein hydrolysate. Regeneration of complete plantlets occurred after the fully developed somatic embryos were transferred
to solidified half-strength MS medium supplemented with 1 mg l−1 gibberellic acid. 相似文献
16.
Cotyledon explants of Panax ginseng produced somatic embryos directly on solid hormone-free MS medium containing 3% (w/v) sucrose while high concentration of NH4NO3 (60 mM) induced embryogenic callus. Ten subcultures of the embryogenic callus on hormone-free MS medium with 40 mM NH4NO3 gave hormone-independent proliferation of callus, which exhibited proliferation potential even on MS medium with a standard level of NH4NO3 (20 mM). Pulse treatment of callus with exogenous auxin or cytokinin (1.0 mg 1–1 2,4-D, 1.0 mg 1–1 kinetin) resulted in the loss of the hormone-independent characteristic and caused the callus to brown. For the suspension culture, embryogenic callus was transferred to MS liquid medium containing 3% (w/v) sucrose in an 500 ml Erlenmyer flask. Embryogenic cell clumps in full-strength MS liquid medium discharged toxic substances, resulting in strong suppression of cell growth. In 1/3-strength MS medium, exudation of toxic material did not occur. Embryogenic cell clumps were mass-grown on a large-scale in a bioreactor (20-1), showing a 7.1 increase of fresh weight in 1/3-strength MS medium with 3% (w/ v) sucrose after 5 weeks of culture. Total ginsenoside content of cultured embryogenic cell clumps was low and 6 times below naturally-cultivated ginseng roots. 相似文献
17.
Hairy roots of maize were induced by infecting 15-d calli with Agrobacterium rhizogenes. The hairy roots cultured in hormone-free media showed the vigorous growth and typical hairy root features. The regenerated
plants were produced from hairy roots in MS media supplemented with 1.6 mg/L ZT and 0.4 mg/L NAA. The PCR-Southern hybridization
demonstrated that T-DNA had been integrated into the chromosome of regenerated plants. 相似文献
18.
Bimal Kumar Ghimire Chang Yeon Yu Ill-Min Chung 《Plant Cell, Tissue and Organ Culture》2012,108(3):455-464
A simple and efficient procedure was developed for in vitro propagation of Solanum aculeatissimum Jacq. using leaf and petiole explants cultured on Murashige and Skoog (MS) medium supplemented with α-naphthalene acetic
acid (NAA) and 6-benzyladenine (BA). Effects of various plant growth regulators, explant types, carbohydrates, and basal salts
on induction of adventitious shoots were also studied. Leaf explants appeared to have better regeneration capacity than petiole
explants in the tested media. The highest regeneration frequency (79.33 ± 3.60%) and shoot number (11.33 ± 2.21 shoots per
explant) were obtained in leaf explants in MS medium containing 3% sucrose and 0.8% agar, supplemented with 0.1 mg/l NAA and
2.0 mg/l BA, whereas petiole explants were more responsive to 0.1 mg/l NAA and 1.0 mg/l thiadiazuron. Developed shoots rooted
best on MS medium with 1.0 mg/l indole acetic acid (IAA), producing 18.33 ± 2.51 roots per shoot. Histological investigation
showed that the shoot buds originated mainly from epidermal cells of wounded tissues, without callus formation. The regenerated
plantlets were successfully acclimatized in a greenhouse, where over 90% developed into morphologically normal and fertile
plants. Results of flow cytometry analysis on S. aculeatissimum indicated no variation in the ploidy levels of plants regenerated via direct shoot formation and showed almost the same phenotype
as that of mother plants. This adventitious shoot regeneration method may be used for large-scale shoot propagation and genetic
engineering studies of S. aculeatissimum. 相似文献
19.
Hairy roots were obtained after inoculation with Agrobacterium rhizogenes strain NCPPB 1855 of the in-vitro-grown shoots of the cherry rootstocks Colt (Prunus avium×P. pseudocerasus) and Mazzard F12/1 (P. avium L.). Not all putatively transgenic roots were able to grow in hormone-free medium. Mazzard F12/1 roots, induced with A. rhizogenes, did not differentiate any shoot or embryo, while both somatic embryos and shoots differentiated from the transgenic roots
of Colt in medium containing 1 mg/l 6-benzylaminopurine and 1 mg/l 1-naphthaleneacetic acid. Somatic embryos were capable
of secondary embryogenesis, but few developed into whole plants. DNA hybridization showed both a different number of bands
and signal intensity in each of the five transgenic shoot clones and embryos examined. In a morphogenetic in vitro test, leaf
explants of the transgenic shoot clones showed an increased capacity to differentiate roots, although clones differed in their
sensitivity to the hormone ratio. Clones from the transgenic shoots had not only an increased rooting ability when grown in
vitro but also exhibited various hairy root phenotypes when cultured in vitro and when transferred into the greenhouse.
Received: 12 December 1996 / Revision received: 21 March 1997 / Accepted: 10 May 1997 相似文献
20.
Summary Excised cotyledons from 8-d-old pumpkin (Cucurbita pepo L.) seedlings were inoculated with Agrobacterium rhizogenes and cultured on hormone-free Murashige and Skoog medium. At the site of inoculation, transformed hairy roots were successfully
induced by using wild strains 8196 (mannopine-type) and 15834 (agropine-type). After a subsequent transfer on a solid MS medium
without hormones, roots obtained by transformation with strain 15834 failed to form stable hairy root cultures, while several
hairy root lines were established with strain 8196. Three hairy root lines, Cp1, Cp2, and Cp31, have spontaneously generated
callus with embryo-like structures after more than 3 yr of growth on the solid medium. The callus proliferation was more frequent
when the autoclaving of nutrient medium, pH 5.7, was prolonged to 30 min. Separated calluses continued to proliferate and
generated embryos with abnormal morphology. The combination of indole-3-acetic acid and benzyladenine had a favorable influence
on embryogenesis and organogenesis in the Cp31 callus line. The Southern analysis of Cp31 root and embryo DNA confirmed the
presence of the T-DNA of Agrobacterium rhizogenes. 相似文献