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1.
A genetic transformation system has been developed for callus cells of Crataegus
aronia using Agrobacterium
tumefaciens. Callus culture was established from internodal stem segments incubated on Murashige and Skoog (MS) medium supplemented with
5 mg l−1 Indole-3-butyric acid (IBA) and 0.5 mg l−1 6-benzyladenine (BA). In order to optimize the callus culture system with respect to callus growth and coloration, different
types and concentrations of plant growth regulators were tested. Results indicated that the best average fresh weight of red
colored callus was obtained on MS medium supplemented with 2 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.5 mg l−1 kinetin (Kin) (callus maintenance medium). Callus cells were co-cultivated with Agrobacterium harboring the binary plasmid pCAMBIA1302 carrying the mgfp5 and hygromycin phosphotransferase (hptII) genes conferring green fluorescent protein (GFP) activity and hygromycin resistance, respectively. Putative transgenic calli
were obtained 4 weeks after incubation of the co-cultivated explants onto maintenance medium supplemented with 50 mg l−1 hygromycin. Molecular analysis confirmed the integration of the transgenes in transformed callus. To our knowledge, this
is the first time to report an Agrobacterium-mediated transformation system in Crataegus
aronia. 相似文献
2.
Ana Sofia Duque Susana de Sousa Araújo Matilde Ataíde Cordeiro Dulce Maria Santos Manuel Pedro Fevereiro 《Plant Cell, Tissue and Organ Culture》2007,90(3):325-330
We developed an alternative methodology for in vitro selection of transgenic Medicago truncatula cv. Jemalong plants using a bifunctional construct in which the coding sequences for the green fluorescent protein (GFP)
and the β-glucuronidase protein (GUS) are fused. An Agrobacterium-mediated transformation protocol was used followed by regeneration via somatic embryogenesis in the dark, to avoid the synthesis and the consequent autofluorescence of chlorophyll. This method
is a clear advantage over antibiotic and herbicide selection in which survival of non-transformed tissue is commonly reported,
with the reassurance that all the somatic embryos selected as GFP positive are transformed. This was subsequently corroborated
by the detection of GUS activity in leaves, stems and roots of the regenerated plants. Without antibiotic selection, and performing
the embryo induction in the dark, it was possible to attest the advantage of using GFP as an in vivo detectable reporter for
early embryo selection. The fusion with the GUS coding sequence provided additional evidence for the transformation of the
previously selected embryos. 相似文献
3.
E. A. Popowich A. P. Firsov T. Y. Mitiouchkina V. L. Filipenya S. V. Dolgov V. N. Reshetnikov 《Plant Cell, Tissue and Organ Culture》2007,90(3):237-244
Transgenic plants of hyacinth (Hyacinthus orientalis L.) cvs. Edisson and Chine Pink have been obtained by Agrobacterium-mediated transformation. Leaf explants of the both hyacinth cultivars regenerated shoots on MS medium containing 2.2 μM BAP
and 0.3 μM NAA at a frequency of 95%. A. tumefaciens strain CBE21 carrying binary vector pBIThau35 was used for transformation. Plasmid pBIThau35 has been produced by cloning
preprothaumatin II cDNA into pBI121 instead of uidA gene. Inoculated leaf explants formed calli and shoots at high frequency on selective medium with 100 mg l−1 kanamycin. Four hyacinth transgenic lines of cv. Chine Pink and one line of cv. Edisson have been selected on medium containing 200 mg l−1 kanamycin. The insertion of thaumatin II gene into hyacinth genome has been confirmed by PCR-analysis. All transgenic plants
expressed substantial amounts of thaumatin II (between 0.06 and 0.28% of the total soluble protein). Hyacinth transgenic lines
were assayed for resistance to the pathogenic fungi Fusarium culmorum and Botrytis cinerea. There were no significant differences between nontransformed control and transgenic leaves of both cultivars. At the same
time the bulbs of the transgenic line Н7401 cv. Chine Pink showed the higher level of resistance to B. cinerea, the bulbs of the transgenic line Н7404 were more resistant to F. culmorum. In both cases the signs of the fungal disease were developed more slowly. The resistance of the bulbs cv. Edisson line to these fungi was not changed. All transgenic hyacinth plant were successfully transferred to soil for further evaluation. 相似文献
4.
The objective of this article is to study the effect of 5-aminolevulinic acid (ALA) and enhanced chlorophyll content, antioxidative
enzymes and photosynthesis rate by foliar application of ALA. We evaluated three concentrations (control-distilled water,
T1-50 mg l−1, T2-150 mg l−1, T3-250 mg l−1) of ALA and seven cultivars, “Sanchidaye” (Sa-1), “Lichuandasuomian” (Li-1), “Aijiaohuang” (Ai-1), “Qingyou” No. 4 (Qi-1),
“Aikang” No. 5 (Ak-1), “Hanxiao” (Ha-1) and “Shulv” (Sl-1). “Ak-1” showed strongest response of POD (peroxidase) enzyme activity
(0.4 U g−1 min−1) in 250 mg l−1 ALA solution. The highest CAT (catalase) activity (0.8 U g−1 min−1) after administration of 250 mg l−1 ALA was observed in “Li-1”. Meanwhile, highest (1.42 mg l−1) total chlorophyll content was also observed in “Ak-1”, when leaves were treated in 50 mg l−1 ALA, “Li-1” and “Ai-1” showed strongest response of specific activity of superoxide dismutase (SOD) in 50 mg l−1 and 50 mg l−1 ALA. Two hundred and fifty milligram per milliliter of ALA-treatment significantly improved the net photosynthetic rate. 相似文献
5.
6.
Rocío Torreblanca Sergio Cerezo Elena Palomo-Ríos José A. Mercado Fernando Pliego-Alfaro 《Plant Cell, Tissue and Organ Culture》2010,103(1):61-69
Olive tree, Olea europaea L., is one of the most commercially important oil crops. A reliable protocol for the genetic transformation of this species
has been developed. Embryogenic calli were infected with different Agrobacterium tumefaciens strains harboring pBINUbiGUSint or pGUSINT binary plasmids. These vectors contain the nos-nptII and the uidA gene driven by the maize polyubiquitin Ubi1 and CaMV35S promoter, respectively. Inoculated explants were cocultured for 2 days, and later selected in the presence of 200 mg l−1 paromomycin. The inclusion of a 3 weeks selection period in liquid medium supplemented with 50 mg l−1 paromomycin was critical for elimination of chimaeric calli. Agrobacterium strain AGL1 containing pBINUbiGUSint plasmid yielded higher transformation frequencies than EHA105 or LBA4404. Globular somatic
embryos (SE), 1–2 mm diameter, cultured in the selection medium in groups of three, were the best explant for transformation.
Using this protocol, transformation frequencies in the range of 20–45%, based on the number of infected explants proliferating
in the selection medium, have been obtained. More than 100 independent transgenic lines were generated, and 16 of them converted
to plants. Transgenic plants were acclimated and grown in the greenhouse, being phenotypically similar to wild type plants.
The uidA gene was strongly expressed in transgenic material during the in vitro regeneration phase; however, β-glucuronidase (GUS)
activity in pBINUbiGUSint transgenic plants was neither detected in shoots growing in vitro nor in acclimated plants. Transgenic
leaves, however, contained high levels of NPTII protein. By contrast, plants transformed with the pGUSINT plasmid showed a
strong GUS activity in leaves. The protocol here described will allow the genetic improvement of this traditional crop. 相似文献
7.
Hye Jin Choi Thummala Chandrasekhar Hyo-Yeon Lee Kyung-Moon Kim 《Plant Cell, Tissue and Organ Culture》2007,91(3):235-242
Transgenic herbicide-resistant sweet potato plants [Ipomoea batatas (L.) Lam.] were produced through Agrobacterium-mediated transformation system. Embryogenic calli derived from shoot apical meristems were infected with Agrobacterium tumefaciens strain EHA105 harboring the pCAMBIA3301 vector containing the bar gene encoding phosphinothricin N-acetyltransferase (PAT) and the gusA gene encoding β-glucuronidase (GUS). The PPT-resistant calli and plants were selected with 5 and 2.5 mg l−1 PPT, respectively. Soil-grown plants were obtained 28–36 weeks after Agrobacterium-mediated transformation. Genetic transformation of the regenerated plants growing under selection was demonstrated by PCR,
and Southern blot analysis revealed that one to three copies of the transgene were integrated into the plant genome of each
transgenic plant. Expression of the bar gene in transgenic plants was confirmed by RT-PCR and application of herbicide. Transgenic plants sprayed with Basta containing
900 mg l−1 of glufosinate ammonium remained green and healthy. The transformation frequency was 2.8% determined by herbicide application
which was high when compared to our previous biolistic method. In addition, possible problems with multiple copies of transgene
were also discussed. We therefore report here a successful and reliable Agrobacterium-mediated transformation of the bar gene conferring herbicide-resistance and this method may be useful for routine transformation and has the potential to develop
new varieties of sweet potato with several important genes for value-added traits such as enhanced tolerance to the herbicide
Basta. 相似文献
8.
Elena Palomo-Ríos Araceli Barceló-Muñoz José A. Mercado Fernando Pliego-Alfaro 《Plant Cell, Tissue and Organ Culture》2012,109(2):201-211
Key factors influencing the efficiency of transformation of embryogenic cultures, induced from immature zygotic embryos, of
avocado cv. ‘Duke 7’ were evaluated. Initially, the sensitivity of somatic embryos to the antibiotics kanamycin, used for
selection, carbenicillin, cefotaxime and timentin, all used for elimination of Agrobacterium cells, were evaluated. Isolated globular somatic embryos were more sensitive to kanamycin than embryogenic masses, and 25 mg l−1 kanamycin completely restricted callus proliferation. Cefotaxime at 500 mg l−1 partially inhibited proliferation of embryogenic cultures, while both carbenicillin and timentin did not affect callus growth.
For genetic transformation, somatic embryos were infected with A. tumefaciens containing the pBINUbiGUSint plasmid. After 2 days, the embryos were transferred to selection medium supplemented with 50 mg l−1 kanamycin and 250 mg l−1 timentin for 2 months. Then, kanamycin level was increased to 100 mg l−1 for two additional months. The A. tumefaciens strain AGL1 yielded higher transformation rates, 6%, than EHA105 or LBA4404, 1.2%. The percentage of kanamycin resistant
calli obtained was significantly influenced by the embryogenic line used as source of explants. Genetic transformation was
confirmed by PCR and Southern blot analysis. A significant improvement in the germination rate was obtained when transgenic
embryos were cultured in liquid MS medium with 4.44 μM BA and 2.89 μM GA3 for 3 days in a roller drum and later transferred to the same medium gelled with 7 g l−1 agar. Plants from five independent transgenic lines were acclimated and grown in the greenhouse, being phenotipically similar
to control plants. 相似文献
9.
A protocol for Agrobacterium-mediated transformation was developed for embryogenic callus of an excellent climber species, Parthenocissus tricuspidata. A. tumefaciens strain EHA105 or C58 harboring the pCAMBIA2301 binary vector with the neomycin phosphotransferase (nptII) and β-glucuronidase (uidA) gene was used. Factors affecting the transformation efficiency, including the Agrobacterium strains, co-cultivation time, Agrobacterium concentration, and infection time, were evaluated. Strain EHA105 proved to be significantly better than C58, and 4 days of
co-culture was critical for transformation. An Agrobacterium suspension at a concentration of 0.5–0.7 × 108 cells ml−1 (OD600 = 0.5–0.7) and an infection time of 40 min was optimal for transformation. By applying these optimized parameters, we recovered
six independent transformed shoots that were kanamycin-resistant and contained the nptII gene, as verified by polymerase chain reaction (PCR) analysis. Southern blot analysis confirmed that T-DNA was stably integrated
into the genome of three out of six PCR-positive lines. Furthermore, histochemical GUS assay revealed the expression of the
uidA gene in kanamycin-resistant calli, somatic embryos, and leaves of transgenic plants. 相似文献
10.
A protocol for Agrobacterium-mediated transformation was developed for in vitro leaf explants of an elite, mature Prunus serotina tree. Agrobacterium tumefaciens strain EHA105 harboring an RNAi plasmid with the black cherry AGAMOUS (AG) gene was used. Bacteria were induced for 12 h with 200 μM acetosyringone for vir gene induction before leaf explant inoculation. Explants were co-cultured for 3 days, and then cultured on woody plant medium
supplemented with 9.08 μM thidiazuron, 1.07 μM napthaleneacetic acid, 60 μM silver thiosulphate, 3% sucrose, plus 200 mg l−1 timentin in darkness for 3 weeks. Regenerating shoots were selected 27 days after initial co-culture, on Murashige and Skoog
medium with 3% sucrose, 8.88 μM 6-benzylaminopurine, 0.49 μM indole-3-butyric acid, 0.29 μM gibberellic acid, 200 mg l−1 timentin, and 30 mg l−1 kanamycin for five subcultures. After 5–6 months of selection, transformation efficiencies were determined, based on polymerase
chain reaction (PCR) analysis of individual putative transformed shoots relative to the initial number of leaf explants tested.
The transformation efficiency was 1.2%. Southern blot analysis of three out of four PCR-positive shoots confirmed the presence
of the neomycin phosphotransferase and AG genes. Transgenic shoots were rooted (37.5%), but some shoot tips and leaves deteriorated or died, making acclimatization
of rooted transgenic plants difficult. This transformation, regeneration, and rooting protocol for developing transgenic black
cherry will continue to be evaluated in future experiments, in order to optimize the system for several mature black cherry
genotypes. 相似文献
11.
Gunvant Patil Amit Deokar P. K. Jain R. J. Thengane R. Srinivasan 《Plant cell reports》2009,28(11):1669-1676
To develop an alternative genetic transformation system that is not dependent on an antibiotic selection strategy, the phosphomannose
isomerase gene (pmi) system was evaluated for producing transgenic plants of chickpea (Cicer arietinum L.). A shoot morphogenesis protocol based on the thidiazuron (TDZ)-induced shoot morphogenesis system was combined with Agrobacterium-mediated transformation of the pmi gene and selection of transgenic plants on mannose. Embryo axis explants of chickpea cv. C-235 were grown on a TDZ-supplemented
medium for shoot proliferation. Embryo axis explants from which the first and second flush of shoots were removed were transformed
using Agrobacterium carrying the pmi gene, and emerging shoots were allowed to regenerate on a zeatin-supplemented medium with an initial selection pressure of
20 g l−1 mannose. Rooting was induced in the selected shoots on an indole-3-butyric acid (IBA)-supplemented medium with a selection
pressure of 15 g l−1 mannose. PCR with marker gene-specific primers and chlorophenol red (CPR) assay of the shoots indicated that shoots had been
transformed. RT-PCR and Southern analysis of selected regenerated plants further confirmed integration of the transgene into
the chickpea genome. These positive results suggest that the pmi/mannose selection system can be used to produce transgenic plants of chickpea that are free from antibiotic resistance marker
genes. 相似文献
12.
Diane E. Darlington Chiu-Yueh Hung Jiahua Xie 《Plant Cell, Tissue and Organ Culture》2009,99(2):157-165
Agrobacterium
tumefaciens strain LBA4404 containing the plasmid pBI121, carrying the reporter gene uidA and the kanamycin resistance gene nptII, was used for gene transfer experiments in selenium (Se)-hyperaccumulator Astragalus racemosus. The effects of kanamycin on cell growth and division and acetosyringone on transformation efficiency were evaluated. The
optimal concentration of kanamycin that could effectively inhibit cell growth and division in non-transgenic tissues was 50 mg l−1 and thus all putative transgenic plants were obtained on induction medium containing 50 mg l−1 kanamycin. The verification of transformants was achieved by both histochemical GUS assay and PCR amplification of nptII gene. Southern blot analysis was performed to further confirm that transgene nptII was stably integrated into the A. racemosus genome. A transformation frequency of approximately 10% was achieved using this protocol, but no beneficial effect from the
addition of acetosyringone (50 μM) was observed. This transformation system will be a useful tool for future studies of genes
responsible for Se-accumulation in A. racemosus. 相似文献
13.
A protocol was developed for rapid and efficient production of transgenic celery plants via somatic embryo regeneration from
Agrobacterium tumefaciens- inoculated leaf sections, cotyledons and hypocotyls. These explants were excised from in vitro seedlings of the cvs. XP166
and XP85 and inoculated with A. tumefaciens strain EHA105 containing the binary vector pBISN1. PBISN1 has the neomycin phosphotransferase gene (nptII) and an intron interrupted β-glucuronidase (GUS) reporter gene (gusA). Co-cultivation was carried out for 4 d in the dark on callus induction medium (CIM): Gamborg B5 + 2.79 μM kinetin + 2.26 μM
2,4-dichlorophenoxyacetic acid (2,4-D) supplemented with 100 μM acetosyringone. Embryogenic calluses resistant to kanamycin
(Km) were then recovered on CIM + 25 mg l−1 Km + 250 mg l−1 timentin after 12 weeks. Subsequently, a large number of Km-resistant and GUS-positive transformants, tens to hundreds per
explant were regenerated via somatic embryogenesis on Gamborg B5 + 4.92 μM 6 (γ,γ-dimethylallylamino)-purine (2iP) + 1.93 μM
α-naphthaleneacetic acid (NAA) + 25 mg l−1 Km + 250 mg l−1 timentin after 8 weeks. Using this protocol, the transformation frequency was 5.0% and 5.0% for leaf sections, 17.8% and
18.3% for cotyledons, and 15.9% and 16.7% for hypocotyl explants of cvs. XP85 and XP166, respectively. Stable integration
of the model transgenes with 1–3 copy numbers was confirmed in all ten randomly selected transgenic events by Southern blot
analysis of gusA. Progeny analysis by histochemical GUS assay showed stable Mendelian inheritance of the transgenes. Thus, A. tumefaciens-mediated transformation of cotyledons or hypocotyls provides an effective and reproducible protocol for large-scale production
of transgenic celery plants. 相似文献
14.
J. F. Liu X. F. Wang Q. L. Li X. Li G. Y. Zhang M. G. Li Z. Y. Ma 《Plant Cell, Tissue and Organ Culture》2011,106(2):207-214
Transgenic cotton with an increased level of phytase activity was generated from cotton (Gossypium hirsutum L.) cv. ND94-7 by subjecting shoot-apex explants to particle bombardment. These tissues were transformed with plasmid pC-KSA2300
carrying a selectable marker (for kanamycin) and a target gene (phytase, or phyA, from Aspergillus ficuum). Primary plants were regenerated in a medium containing 75 mg l−1 kanamycin. Of 1,534 shoot apices, 52 (3.4%) survived on this selection medium. Southern and Northern blot analyses confirmed
that phyA was stably integrated and expressed in those primary transgenics. The progenies of the primary transgenic plants were found
to have a 3.1- to 3.2-fold increase in root extracellular phytase activity, resulting in improved phosphorus (P) nutrition.
Growth also was enhanced when they were supplied with phytate, and their P content was equivalent to that of wildtype plants
supplied with inorganic phosphate. These results demonstrate that the expression of phyA in cotton plants improves their ability to utilize organic P in response to a deficiency. 相似文献
15.
Rangaraj Nandakumar Li Chen Suzanne M. D. Rogers 《In vitro cellular & developmental biology. Plant》2007,43(3):187-194
A highly reproducible Agrobacterium-mediated transformation system was developed for the wetland monocot Juncus accuminatus. Three Agrobacterium tumefaciens binary plasmid vectors, LBA4404/pTOK233, EHA105/pCAMBIA1201, and EHA105/pCAMBIA1301 were used. All vectors contained the
35SCaMV promoter driven, intron containing, β-glucuronidase (gus), and hygromycin phosphotransferase (hptII) genes within their T-DNA. After 48 h of cocultivation, 21-d-old seedling derived calli were placed on medium containing
timentin at 400 mg l−1, to eliminate the bacteria. Calli were selected on MS medium containing 40 or 80 mg l−1 hygromycin, for 3 mo. Resistant calli were regenerated and rooted on MS medium containing hygromycin, 5 mg l−1(22.2 μM) of 6-benzylamino-purine (BA) and 0.1 mg l−1(0.54 μM) of alpha-naphthaleneacetic acid (NAA), respectively. Seventy-one transgenic cell culture lines were obtained and
39 plant lines were established in the greenhouse. All the plants were fertile, phenotypically normal, and set viable seed.
Both transient and stable expression of the gus gene were demonstrated by histochemical GUS assays of resistant calli, transgenic leaf, root, inflorescence, seeds, and whole
plants. The integration of gus and hptII genes were confirmed by polymerase chain reaction (PCR) and Southern analysis of both F0 and F1 progenies. The integrated genes segregated to the subsequent generation in Mendelian pattern. To our knowledge, this is the
first report of the generation of transgenic J. accuminatus plants. 相似文献
16.
17.
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
18.
Meiru Li Hongqing Li Huawu Jiang Xiaoping Pan Guojiang Wu 《Plant Cell, Tissue and Organ Culture》2008,92(2):173-181
Jatropha curcas contains high amounts of oil in its seed and has been considered for bio-diesel production. A transformation procedure for
J. curcas has been established for the first time via
Agrobacterium tumefaciens infection of cotyledon disc explants. The results indicated that the efficiency of transformation using the strain LBA4404
and phosphinothricin for selection was an improvement over that with the strain EHA105 and hygromycin. About 55% of the cotyledon
explants produced phosphinothricin-resistant calluses on Murashige and Skoog (MS) medium supplemented with 1.5 mg l−1 benzyladenine (BA), 0.05 mg l−1 3–indolebutyric acid (IBA), 1 mg l−1 phosphinothricin and 500 mg l−1 cefotaxime after 4 weeks. Shoots were regenerated following transfer of the resistant calli to shoot induction medium containing
1.5 mg l−1 BA, 0.05 mg l−1 IBA, 0.5 mg l−1 gibberellic acid (GA3), 1 mg l−1 phosphinothricin and 250 mg l−1 cefotaxime, and about 33% of the resistant calli differentiated into shoots. Finally, the resistant shoots were rooted on
1/2 MS media supplemented with 0.3 mg l−1 IBA at a rate of 78%. The transgenic nature of the transformants was demonstrated by the detection of β-glucuronidase activity
in the primary transformants and by PCR and Southern hybridization analysis. 13% of the total inoculated explants produced
transgenic plants after approximately 4 months. The procedure described will be useful for both, the introduction of desired
genes into J. curcas and the molecular analysis of gene function. 相似文献
19.
Saba Ambreen Memon Xilin Hou Bo Zhu Joseph N. Wolukau 《Acta Physiologiae Plantarum》2009,31(6):1191-1196
The objective of the present work was selection of cultivar and suitable medium for regenerating shoots from leaf segments
of non-heading Chinese cabbage. We evaluated six types of supplemented media with 2.0, 5.0 and 10.0 mg l−1 6-BA; 1.0 and 2.0 mg l−1 TDZ; 0.1, 0.3, 0.5, 0.8 and 1.0 mg l−1NAA; 3.0, 5.0 and 7.5 mg l−1AgNO3; 0.01 mg l−1 2–4, D and 4.0 mg l−1 KT for shoot regeneration and six cultivars “Sanchidaye”, “Liuchuandasuomian”, “Qingyou 4”, “Liangbaiye”, “AiKang 5” and
“Hanxiao F3”, furthermore for root formation three types of supplemented media with 0.2, 0.3, 0.5 mg l−1 NAA, and for survival rate two types of base media: turf + vermiculite + manure (1:2:0.2) and soil + vermiculite (1:2). Culturing
leaf segments on MS medium supplemented with 2 mg l−1 TDZ; 0.5 mg l−1 NAA and 7.5 mg l−1 AgNO3 gave the highest number of shoots per leaf segment (66) while roots were best formed on the medium supplemented with 0.2 mg l−1 NAA. Survival rate was highest (61.6%) in the turf: vermiculite: manure (1:2:0.2) medium. The highest percentage of responding
leaf segments, number of shoots per leaf segment, rooting percentage and survival rate were observed in “Liuchuandasuomian”.
The plantlets were transferred to the soil and grown into mature plants in pots. These results could be used for preliminary
selections of cultivars to transfer disease resistance (Bt) gene through agrobacterium in non-heading Chinese cabbage. 相似文献
20.
Bo Yu Hong Zhai Yuping Wang Ning Zang Shaozhen He Qingchang Liu 《Plant Cell, Tissue and Organ Culture》2007,90(3):265-273
Efficient Agrobacterium tumefaciens-mediated transformation was achieved using embryogenic suspension cultures of sweetpotato (Ipomoea batatas (L.) Lam.) cv. Lizixiang. Cell aggregates from embryogenic suspension cultures were cocultivated with the A. tumefaciens strain EHA105 harboring a binary vector pCAMBIA1301 with gusA and hygromycin phosphotransferase II gene (hpt II) genes. Selection culture was conducted using 25 mg l−1 hygromycin. A total of 2,218 plants were regenerated from the inoculated 1,776 cell aggregates via somatic embryogenesis.
β-glucuronidase (GUS) assay and PCR, dot blot and Southern blot analyses of the regenerated plants randomly sampled showed
that 90.37% of the regenerated plants were transgenic plants. The number of integrated T-DNA copies varied from 1 to 4. Transgenic
plants, when transferred to soil in a greenhouse and a field, showed 100% survival. No morphological variations were observed
in the ex vitro transgenic plants. These results exceed all transformation experiments reported so far in the literature in
quantity of independent events per transformation experiment in sweetpotato. 相似文献