共查询到20条相似文献,搜索用时 515 毫秒
1.
Transgenic Podophyllum peltatum plants were successfully produced by Agrobacterium tumefaciens-mediated transformation. Embryogenic callus was co-cultivated with Agrobacterium tumefaciens harboring a binary vector pBI 121 carrying β-glucuronidase (GUS) and neomycinphosphotransferase (NPT II) gene. GUS-histochemical analysis revealed that, 50 μM acetosyringone treatments during Agrobacterium infection and 3 d co-cultivation with Agrobacterium showed enhanced transformation efficiency. Percentage of GUS positive callus increased rapidly as the subculture time proceeded
on selection medium containing 100 mg dm−3 kanamycin. Kanamycin resistant somatic embryos were formed from embryogenic callus after cultivation with 11.35 μM abscisic
acid (ABA) for 3 weeks and then on hormone-free selection medium. Somatic embryos were germinated and converted into plantlets
on medium containing 2.89 μM gibberellic acid (GA3). The integration of GUS and NPT II gene into transgenic plants was confirmed by polymerase chain reaction and Southern analysis. 相似文献
2.
Embryogenic calli were induced from leaf explants of coffee (Coffea canephora) on McCown's woody plant medium (WPM) supplemented with 5 μM N6–(2-isopentenyl)-adenosine (2-iP). These calli were co-cultured with Agrobacterium tumefaciens EHA101 harboring pIG121-Hm, containing β-glucuronidase (GUS), hygromycin phosphotransferase (HPT), and neomycin phosphotransferase
II genes. Selection of putative transgenic callus was performed by gradual increase in hygromycin concentration (5, 50, 100 mg/l).
The embryogenic calli surviving on medium containing 100 mg/l hygromycin showed a strong GUS-positive reaction with X-Gluc
solution. Somatic embryos were formed from these putative transgenic calli and germinated on WPM medium with 5 μM 2-iP. Regenerated
small plantlets with shoots and roots were transferred to medium containing both 100 mg/l hygromycin and 100 mg/l kanamycin
for final selection of transgenic plants. The selected plantlets exhibited strong GUS activity in leaves and roots as indicated
by a deep blue color. GUS and HPT genes were confirmed to be stably integrated into the genome of the coffee plants by the
polymerase chain reaction.
Received: 14 December 1998 / Revision received: 12 March 1999 / Accepted: 24 March 1999 相似文献
3.
Hairy roots of Panax ginseng were obtained after root disks were infected with wild-type strain Agrobacterium rhizogenes 15834. Three lines of hairy roots with different pigmentation were selected. Embryogenic callus was induced on Murashige
and Skoog medium containing 1.0 mg/l 2,4-D. The frequency of embryogenic callus formation was 37.4% in a line with red pigmentation.
Somatic embryo development from embryogenic callus was efficiently achieved by lowering the concentration of 2,4-D (0.5 mg/l).
After the germination of somatic embryos on medium with 10 mg/l GA3, the embryos were transferred to 1/2-MS medium without GA3. The transformed ginseng plantlets had an actively growing root system with abundant lateral roots. The phenotypical alteration
of transformed ginseng plants might be valuable character for increasing root yield.
Received: 27 March 1999 / Revision received: 18 May 1999 / Accepted 8 July 1999 相似文献
4.
Sedums are used as groundcover, in rock gardens and flower borders, and for greening the top floor of buildings, cottages, and thatched roofs. In this study, Agrobacterium-mediated genetic transformation of Sedum erythrostichum was studied by introducing a herbicide-resistant gene (phosphinothricin-N-acetyl-transferase) and a reporter gene (#-glucuronidase, GUS). Following co-cultivation with Agrobacterium on MS medium supplemented with 0.5 mg/l !-naphthaleneacetic acid (NAA) and 2 mg/l 6-benzylaminopurine (BA) for 3 days, leaf segments were transferred onto medium containing 300 mg/l cefotaxime. When adventitious shoots developed directly near the margins of explants after 3 weeks, they were transferred to selection medium with 25 mg/l kanamycin. Of a total of 640 infected leaf explants, 24 (3.75%) produced kanamycin-resistant adventitious shoots; of these, 2.5% were GUS-positive. Transgenic plantlets were confirmed using polymerase chain reaction, Southern, and Northern analyses. Ninety-four percent of the transgenic plantlets were successfully transferred to soil and produced flowers. All GUS-positive transgenic plants were strongly resistant to Basta (phosphinothricin at 200 mg/l) after spraying. 相似文献
5.
Plantlets were regenerated from cultured seed explants of the forage grass Caucasian bluestem [Bothriochloa caucasica (Trin.) C.E. Hubbard] via somatic embryogenesis. Embryogenic callus was produced in four weeks when surface sterilized seeds were cultured on a medium containing MS-salts, B-5 vitamins, 12 mM L-proline, 2% sucrose, 0.8% agar and 5M 2,4-D. Plantlets were regenerated in 6–8 weeks after culture initiation. Healthy root and shoot systems were produced within three weeks after the plantlets were transferred to a medium lacking 2,4-D. Approximately 95% of the plantlets survived greenhouse acclimation and produced healthy plants and viable seeds. Caucasian bluestem callus cultures exhibit natural resistance to kanamycin. High levels of kanamycin (up to 800 mg/l) did not completely inhibit callus growth. However, the regeneration of healthy-plantlets was completely inhibited by kanamycin even at low levels (50 mg/l). 相似文献
6.
Genetic transformation of the actinorhizal tree Allocasuarina verticillata by Agrobacterium tumefaciens 总被引:1,自引:0,他引:1
C. Franche D. Diouf Q.V. Le D. Bogusz A. N'Diaye H. Gherbi C. Gobé E. Duhoux 《The Plant journal : for cell and molecular biology》1997,11(4):897-904
We have developed an efficient transformation system for the tropical actinorhizal tree Allocasuarina verticillata using Agrobacterium tumefaciens mediated gene transfer. Mature zygotic embryos were inoculated with the disarmed strain C58C1 carrying, in the binary vector BIN19, the nptll gene, providing kanamycin resistance as a selectable marker, and the reporter gene β-glucuronidase containing an intron. The transformed embryos were cultivated on nutrient medium supplemented with 0.5 µM NAA, 2.5 µM BA, 100 mg l−1 kanamycin and 250 mg l−1 cefotaxime. After 2 months, a 21% transformation frequency was obtained. Within 6–9 months, transgenic plants were recovered from 70% of the transformed calli. The presence of the transgenes was demonstrated by PCR analysis and by the expression of the β-glucuronidase; integration of the T-DNA was confirmed by Southern hybridization. More than 100 transgenic plants from a total of 23 independent transformation events have been successfully established in soil. The possibility to obtain nitrogen-fixing nodules after inoculation of transgenic A. verticillata plants by the actinomycetal strain of Frankia Allo2 was established. 相似文献
7.
An efficient variety-independent method for producing transgenic eggplant (Solanum melongena L.) via Agrobacterium tumefaciens-mediated genetic transformation was developed. Root explants were transformed by co-cultivation with Agrobacterium tumefaciens strain LBA4404 harbouring a binary vector pBAL2 carrying the reporter gene beta-glucuronidase intron (GUS-INT) and the marker gene neomycin phosphotransferase (NPTII). Transgenic calli were induced in media containing 0.1 mg l(-1) thidiazuron (TDZ), 3.0 mg l(-1) N(6)-benzylaminopurine, 100 mg l(-1) kanamycin and 500 mg l(-1) cefotaxime. The putative transgenic shoot buds elongated on basal selection medium and rooted efficiently on Soilrite irrigated with water containing 100 mg l(-1) kanamycin sulphate. Transgenic plants were raised in pots and seeds subsequently collected from mature fruits. Histochemical GUS assay and polymerase chain reaction analysis of field-established transgenic plants and their offsprings confirmed the presence of the GUS and NPTII genes, respectively. Integration of T-DNA into the genome of putative transgenics was further confirmed by Southern blot analysis. Progeny analysis of these plants showed a pattern of classical Mendelian inheritance for both the NPTII and GUS genes. 相似文献
8.
Xiang Ling You Xiao Tan Jin Ling Dai Yu Hua Li Yong Eui Choi 《Plant Cell, Tissue and Organ Culture》2012,108(2):333-338
An efficient system for inducing somatic embryogenesis in Panax notoginseng was established using shaker flasks and bioreactor cultures; furthermore, regenerated plantlets were successfully transferred
to ex vitro soil conditions. Embryogenic callus was induced from segments of adventitious roots incubated on Murashige and
Skoog (MS) medium containing 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) after 5 weeks of culturing. The highest frequency
(100%) of somatic embryogenesis, with a mean of 32.7 somatic embryos per callus, was obtained on embryogenic callus incubated
on a medium containing 0.5 mg/L 2,4-D. To scale-up somatic embryo formation, 10 g (~1.65 × 104) of early globular-stage somatic embryos were incubated in a 3 L airlift bioreactor containing 1.5 L 1/2 MS medium without
plant growth regulators (PGRs) for a period of 4 weeks; these globular-stage somatic embryos then developed into cotyledonary
embryos. When maintained on PGR-free medium, the cotyledonary embryos developed roots but did not develop shoots. However,
when they were treated with gibberellic acid (GA3), they continued to germinate and transformed into plantlets after 2 weeks of culture. Plantlets with well-developed shoots
and roots were transferred to an autoclaved vermiculite and perlite mixture, acclimatized for a period of 3 months and successfully
transferred to forest mountain soil. Following overwintering, these plants produced new growth. 相似文献
9.
Embryogenic callus was induced from leaf explants of Eleutherococcus sessiliflorus cultured on Murashige and Skoog (MS) basal medium supplemented with 1 mg l(-1) 2,4-dichlorophenoxyacetic acid (2,4-D), while no plant growth regulators were needed for embryo maturation. The addition of 1 mg l(-1) 2,4-D was needed to maintain the embryogenic culture by preventing embryo maturation. Optimal embryo germination and plantlet development was achieved on MS medium with 4 mg l(-1) gibberellic acid (GA(3)). Low-strength MS medium (1/2 and 1/3 strength) was more effective than full-strength MS for the production of normal plantlets with well-developed shoots and roots. The plants were successfully transferred to soil. Embryogenic callus was used to establish a suspension culture for subsequent production of somatic embryos in bioreactor. By inoculating 10 g of embryogenic cells (fresh weight) into a 3l balloon type bubble bioreactor (BTBB) containing 2l MS medium without plant growth regulators, 121.8 g mature somatic embryos at different developmental stages were harvested and could be separated by filtration. Cotyledonary somatic embryos were germinated, and these converted into plantlets following transfer to a 3l BTBB containing 2l MS medium with 4 mg l(-1) GA3. HPLC analysis revealed that the total eleutherosides were significantly higher in leaves of field grown plants as compared to different stages of somatic embryo. However, the content of eleutheroside B was highest in germinated embryos. Germinated embryos also had higher contents of eleutheroside E and eleutheroside E1 as compared to other developmental stages. This result indicates that an efficient protocol for the mass production of E. sessiliflorus biomass can be achieved by bioreactor culture of somatic embryos and can be used as a source of medicinal raw materials. 相似文献
10.
Artur da Câmara Machado Monika Puschmann Helene Pühringer Robert Kremen Hermann Katinger Margit Laimer da Câmara Machado 《Plant cell reports》1995,14(6):335-340
Embryogenic lines of Prunus subhirtella autumno rosa were established on a modified MS medium supplemented with 1 mg/l NAA, 0.06 mg/l IBA and 0.04 mg/l BA from petioles of axenically grown shoots of adult origin. To induce normal development of plantlets we compared a range of approaches on solid culture media as well as in suspension cultures including treatments with ABA, GA3, zeatin, darkness, and cold. A series of experiments were conducted to follow the temporal pattern of somatic embryo development.Separation of embryos at different stages of development was carried out by sieving the suspension cultures through nylon nets. While the embryogenic masses were used for further subcultures, well formed embryos were used for germination experiments.Transformed Prunus subhirtella plants were regenerated from somatic embryos by inoculating an embryogenic callus with Agrobacterium strain LBA 4404 containing the ß-glucuronidase (GUS) gene on plasmid pBinGUSint. Several putative transformed embryogenic calli were selected for continued proliferation on kanamycin containing media. Finally transgenic plants were regenerated on shoot multiplication medium containing kanamycin. Embryos and plants were shown to express the GUS gene by histochemical assays and northern blot analysis. With a yield of 110 transgenic lines from a single transformation experiment this approach appears ideal for the study of the influence on level of expression caused by different copy number, site of insertion etc. This will be helpful in establishing parameters according to which the best transgenic line for a chosen purpose should be selected.Abbreviations BA
6-benzylaminopurine
- IBA
3-indolebutyric acid
- GA3
gibberellic acid
- NAA
1-naphthylacetic acid
- ABA
abscisic acid
- GUS
ß-glucuronidase
- NPTII
neomycin phosphotransferase II
- SDS
sodium dodecyl sulphate
- SSC
standard saline citrate
- PEM
proembryogenic masses
Dedicated to Franticek Novak 相似文献
11.
An efficient transformation protocol was developed for Eucalyptus tereticornis Sm. using cotyledon and hypocotyl explants. Precultured cotyledon and hypocotyl explants were cocultured with Agrobacterium tumefaciens strain LBA 4404 harboring the binary vector pBI121 containing the uidA and neomycin phosphotransferase II genes for 2 d and transferred to selective regeneration medium containing 0.5 mg/l 6-benzylaminopurine
(BAP), 0.1 mg/l naphthalene acetic acid, 40 mg/l kanamycin, and 300 mg/l cefotaxime. After two passages in the selective regeneration
medium, the putatively transformed regenerants were transferred to Murashige and Skoog (MS) liquid medium containing 0.5 mg/l
BAP and 40 mg/l kanamycin on paper bridges for further development and elongation. The elongated kanamycin-resistant shoots
were subsequently rooted on the MS medium supplemented with 1.0 mg/l indole-3-butyric acid and 40 mg/l kanamycin. A strong
β-glucuronidase activity was detected in the transformed plants by histochemical assay. Integration of T-DNA into the nuclear
genome of transgenic plants was confirmed by polymerase chain reaction and southern hybridization. This protocol allows effective
transformation and direct regeneration of E. tereticornis Sm. 相似文献
12.
Somatic embryogenesis was obtained from cotyledon and mature zygotic embryo callus cultures of Terminalia chebula Retz. Callus cultures of cotyledon and mature zygotic embryo were initiated on induction medium containing Murashige and Skoog (MS) nutrients with 1.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) either 0.01 or 0.1 mg/l Kinetin and 30 g/l sucrose. Induction of somatic embryogenesis, proliferation and development was obtained through different culture passages. Embryogenic cotyledon callus with globular somatic embryos was obtained on MS basal medium supplemented with 50 g/l sucrose. Globular somatic embryos were observed from mature zygotic embryo callus on induction medium. Different stages of somatic embryo development from cotyledon and mature zygotic embryo calluses were observed on MS basal medium supplemented with 50 g/l sucrose after 4 weeks of culture. Histological studies have revealed the developmental stages of somatic embryos. A maximum of 40.3±1.45 cotyledonary somatic embryos/callus was obtained from mature zygotic embryo compared to 7.70±0.37 cotyledonary somatic embryos/callus initiated from cotyledons. Germination of somatic embryos and conversion to plants were achieved. Highest frequency of germination (46.66±0.88) of somatic embryos was obtained on MS basal medium containing benzyladenine (0.5 mg/l) with 30 g/l sucrose. 相似文献
13.
A protocol was developed for Agrobacterium-mediated genetic transformation of niger [ Guizotia abyssinica (L.f.) Cass.] using hypocotyl and cotyledon explants. Hypocotyls and cotyledons obtained from 7-day-old seedlings were co-cultivated with Agrobacterium tumefaciens strain EHA101/pIG121Hm that harbored genes for beta-glucuronidase (GUS), kanamycin, and hygromycin resistance. Following co-cultivation, the hypocotyl and cotyledon explants were cultivated on MS medium containing 1 mg/l 6-benzylaminopurine (BA) for 3 days in darkness. Subsequently, hypocotyl and cotyledon explants were transferred to selective MS medium containing 1 mg/l BA, 10 mg/l hygromycin, 10 mg/l kanamycin, and 500 mg/l cefotaxime. After 6 weeks, hypocotyls and cotyledons produced multiple adventitious shoot buds, and these explants were subcultured to MS medium containing 1 mg/l BA, 30 mg/l hygromycin, and 30 mg/l kanamycin. After a further 3 weeks, the explants (along with developing shoot buds) were subcultured to MS medium containing 1 mg/l BA, 50 mg/l kanamycin, and 50 mg/l hygromycin for further selection. Transgenic plants were obtained after rooting on half-strength MS medium supplemented with 0.1 mg/l alpha-naphthaleneacetic acid, 50 mg/l kanamycin, and 50 mg/l hygromycin and were confirmed by GUS histochemical assay and polymerase chain reaction analysis. Genomic Southern blot hybridization confirmed the incorporation of the neomycin phosphotransferase II gene into the host genome. 相似文献
14.
Qi Zhu Fengtao Wu Feng Ding Dong Ye Yongqin Chen Yi Li Yang Zhifan 《Plant Cell, Tissue and Organ Culture》2009,96(3):317-324
Dioscorea zingiberensis Wright has been cultivated as a pharmaceutical crop for production of diosgenin, a precursor for synthesis of various important
steroid drugs. Because breeding of D. zingiberensis through sexual hybridization is difficult due to its unstable sexuality and differences in timing of flowering in male and
female plants, gene transfer approaches may play a vital role in its genetic improvement. In this study, the Agrobacterium tumefaciens-mediated transformation of D. zingiberensis was investigated with leaves and calli as explants. The results showed that both leaf segments and callus pieces were sensitive
to 30 mg/l hygromycin and 50–60 mg/l kanamycin, and using calli as explants and addition of acetosyringone (AS) in cocultivation
medium were crucial for successful transformation. We first immersed callus explants in A. tumefaciens cells for 30 min and then transferred the explants onto a co-cultivation medium supplemented with 200 μM AS for 3 days. Three
days after, we cultured the infected explants on a selective medium containing 50 mg/l kanamycin and 100 mg/l timentin for
formation of kanamycin-resistant calli. After the kanamycin-resistant calli were produced, we transferred them onto fresh
selective medium for shoot induction. Finally, the kanamycin resistant shoots were rooted and the stable incorporation of
the transgene into the genome of D. zingiberensis plants was confirmed by GUS histochemical assay, PCR and Southern blot analyses. The method reported here can be used to
produce transgenic D. zingiberensis plants in 5 months and the transformation frequency is 24.8% based on the numbers of independent transgenic plants regenerated
from initial infected callus explants. 相似文献
15.
Somatic embryo formation and germination from immature embryo-derived suspension-cultured cells of Angelica sinensis (Oliv.) Diels 总被引:3,自引:0,他引:3
Embryogenic callus was induced from immature embryos of Angelica sinensis cultured on Murashige and Skoog (MS) basal medium. Embryogenic callus growth was more rapid on MS basal medium than on B5
or White medium. Embryogenic callus was used to establish a suspension culture and somatic embryos and germinating embryos
developed during the culture. A shaking speed of 80 rpm was found to be optimal for establishing suspension cultures, while
100 rpm produced more somatic embryos and germinating embryos with an initiation cell density of 0.2 ml packed cell volume/25
ml medium. Adding 0.3% agar to the liquid medium also stimulated the formation of somatic and germinating embryos. While no
plant growth regulators were needed for culture initiation and plant regeneration, the addition of 0.5–1 mg/l 2,4-dichlorophenoxyacetic
acid was needed to maintain the embryogenic suspension culture by preventing embryo germination. Forty percent of the germinating
embryos survived after culturing on filter paper moistened with liquid half-strength MS medium containing 3% sucrose. The
plants were successfully transferred into soil.
Received: 19 March 1997 / Revision received: 21 November 1997 / Accepted: 19 January 1998 相似文献
16.
Embryogenic callus and suspension cultures of eastern white pine (Pinus strobus) have been obtained. The whole female gametophyte was plated on a medium containing 50 mg/l glutamine, 500 mg/l casein hydrolysate, 3% sucrose, 2 mg/1 2,4-D, 1 mg/1 BA and 0.2% Gelrite as a solidifying agent. Embryogenic calli could be seen as early as 5 days following culture. Histological studies indicate proliferation of pre-existing embryogenic tissue in the corrosion cavity followed by extrusion of embryogenic callus through the micropylar end of the gametophyte. Embryogenic suspension cultures were obtained by placing embryogenic callus into liquid medium. Embryogenic suspension cultures were subcultured weekly and proliferated as early-stage embryos with attached suspensors. Embryo development was obtained following transfer of the embryogenic tissue to an auxin-free medium containing 50 mM glutamine, 38 M abscisic acid, and 6% sucrose. Although embryo development could be consistently obtained, whole plants have not yet been recovered from these somatic embryos.Abbreviations 2,4-D
2,4-Dichlorophenoxyacetic acid
- ABA
Abscisic acid
- BA
6-Benzyladenine
Salaries and research support were provided by State and Federal funds appropriated to OSU/OARDC. Journal Article No. 62–89 相似文献
17.
Jingli Yang Chenguang Zhou Lihui Liu Du Jia Zhiyang Yin Myongjo Kim Changyeon Yu Chenghao Li 《Plant Cell, Tissue and Organ Culture》2012,110(2):289-298
Embryogenic callus was obtained from young petiole, stem, and root explants of 4-year-old Siberian ginseng plants on medium supplemented with 2,4-dichlorophenoxyacetic acid. Embryogenic callus differentiated into somatic embryos (SEs), most of which could germinate but developed abnormally. Friable embryogenic callus was induced mainly from the root regions of germinated primary SEs or regenerated plantlets on plant growth regulator-free medium. Histological studies showed that the embryogenic callus initiated from the subepidermal cells of young roots. The bioreactor system was more efficient than suspension culture regarding the number and growth of SEs, although a similar amount of embryonic tissue was used. An average of 115,370 germinated SEs developed from an initial 400?mg of embryogenic callus, and 64.7?% of germinated SEs converted into plantlets after a 4-weeks culture on agar medium. During the bioreactor culture process, secondary SEs were induced directly from SEs at various stages, a phenomenon that rarely occurred in suspension culture. These secondary SEs developed quickly and germinated during the bioreactor culture process. Proline content and peroxidase and catalase activities of SEs cultured in bioreactors were higher than in SEs cultured in suspension culture. 相似文献
18.
D. H. Tejavathi M. D. Rajanna R. Sowmya K. Gayathramma 《In vitro cellular & developmental biology. Plant》2007,43(5):423-428
Somatic embryogenesis from cultures of shoot apices, cotyledon and young leaves of in vitro shoots of Agave vera-cruz Mill. was studied. Embryogenic callus was obtained when explants were cultured on Murashige and Skoog’s (MS) medium (1962)
supplemented with L2 vitamins, 4.52 μM 2,4-dichlorophenoxyacetic acid (2,4-d) or 5.37 μM ∝-naphthalene acetic acid (NAA). Somatic embryos differentiated from this embryogenic callus upon subculture
to maturation/conversion medium containing cytokinin either alone or with auxin and l-glutamine. The best combination of growth regulators for development of somatic embryos was found to be 5.37 μM naphthalene
acetic acid plus 0.91 μM zeatin and 40 g/l sucrose. The conversion frequency of somatic embryos to plantlets varied from 46–50%.
Rooted plantlets were transferred directly to pots containing a soil, sand, and manure mixture without any hardening phase
with 96–98% survival of the plantlets. Based on the histological observations, the potential origin of the somatic embryo
is discussed. 相似文献
19.
A high-frequency and simple procedure for Agrobacterium tumefaciens-mediated genetic transformation of the medicinal plant Salvia miltiorrhiza was developed. Leaf discs were pre-cultured on MS medium supplemented with 6.6 μmol l−1 BAP and 0.5 μmol l−1 NAA for one day, then co-cultured with A. tumefaciens strain EHA105 harboring the plasmid pCAMBIA 2301 for three days on the same medium. Regenerated buds were obtained on selection
medium (co-culture medium supplemented with 60 mg l−1 kanamycin and 200 mg l−1 cefotaxime) after two cycles’ culture of 10 days each and then transferred to fresh MS medium with 60 mg l−1 kanamycin for rooting. Fifteen days later, the rooted plantlets were obtained and then successfully transplanted to soil.
The transgenic nature of the regenerated plants was confirmed by PCR, Southern hybridization analysis and GUS histochemical
assay. Averagely, 1.1 independent verified transgenics per explant plated were obtained through this protocol. Adopting this
procedure, positive transformed plants could be obtained within 2–3 months from mature seeds germination to transplant to
soil, and more than 1,000 transgenic plants with several engineered constructs encoding different genes of interest were produced
in our lab in the past two years. 相似文献
20.
根癌农杆菌介导GUS基因对白桦转化的研究 总被引:1,自引:0,他引:1
本文报道了根癌农杆菌介导GUS基因转化白桦叶盘的研究方法,对转化过程中各种因素进行了全面探讨,结果表明:白桦叶盘对卡那霉素的敏感实验,得出最适筛选浓度为50mg/l;对头孢霉素和羧苄青霉素的抑菌实验,最适抑菌浓度为500mg/l;100μMAS的加入可将抗性不定芽的诱导率提高3倍;3~4d的预培是最适合叶盘转化的时间;10~20min是最适合叶盘浸染的时间;3~4d的共培养对于叶盘转化比较合理。在上述最适合的转化条件下,农杆菌介导的GUS基因对白桦叶盘的转化获得了抗性再生植株,转化率为2.8%。 相似文献