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Quan-le Xu Zhe Hu Chun-yuan Li Xin-yu Wang Chong-ying Wang 《In vitro cellular & developmental biology. Plant》2009,45(5):583-590
Two protocols were developed for the efficient regeneration of Sinningia speciosa from leaf explants via two developmental pathways. The first method involved formation of callus and buds, followed by subsequent
root growth, in Murashige and Skoog medium (MS) containing 2.0 mg l−1 6-benzylaminopurine (BA) and 0.2 mg l−1 α-naphthalene acetic acid (NAA), with a regeneration efficiency of 99.0%. The second method involved producing callus and
roots, followed by subsequent formation of buds, in MS medium supplemented with 1.0–5.0 mg l−1 NAA, and resulted in a regeneration efficiency of 90.4%. Our experiments indicate that the root-first pathway resulted in
a lower plant regeneration efficiency. Through five continual generations using the buds-first method, a total of 215 regenerated
plants were obtained in the last generation, and eight exhibited a phenotype we named tricussate whorled phyllotaxis (twp). Six of the regenerated twp variant plants maintained their tricussate whorled phyllotaxis phenotype, showing no other abnormalities, while one reverted
to a wild type before flowering and another formed two rounds of sepals. Physiological analysis revealed that the twp plants responded differently than wild type to exogenous NAA and 2,3,5-triiodobenzoic acid (TIBA), while high-performance
liquid chromatography (HPLC) analysis showed that the levels of endogenous indole-3-acetic acid (IAA) and gibberellin (GA)
were lower in twp than wild-type plants. These results suggest that the formation of the twp mutant may be related to phytohormones and that the twp variant could be an important material for investigating the molecular mechanism of plant phyllotaxis patterning. 相似文献
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Xiangbin Xu Jufang Bian Songbai Liu Hongmiao Song Nongnong Shi Yuezhi Tao Huizhong Wang 《Molecular breeding : new strategies in plant improvement》2011,27(3):337-346
The PROMOTION OF CELL SURVIVAL 1 (PCS1) gene, encoding an aspartic protease, has an important role in determining the fate of cells in embryonic development and
reproduction processes in Arabidopsis. To explore the potential function of the PCS1 gene in generating reproductive sterility, we placed the PCS1 gene under the control of an 1,869-bp nucleotide sequence from the 3′ end of the second intron (AG-I) of Arabidopsis AGAMOUS and CaMV 35S (–60) minimal promoter [AG-I-35S (–60)::PCS1], and introduced it into tobacco. RT–PCR results demonstrated that the PCS1 gene driven by AG-I-35S (–60) chimeric promoter was expressed only in anthers and carpels in the reproductive tissues of transgenic tobacco. Compared to
wild-type plants, all AG-I-35S (–60) and AG-I-35S (–60)::PCS1 transgenic lines showed a normal phenotype throughout the vegetative growth phase. However, during the reproductive stage,
most AG-I-35S (–60)::PCS1 transgenic plant anthers displayed delayed dehiscence, failed dehiscence, petalody and hypoplasia, and the pollen grains
had different shapes and sizes with a distorted, shrunken, or collapsed morphology. Moreover, three transgenic lines, PCS1-1,
PCS1-3 and PCS1-4, showed higher sterility than wild-type and AG-I-35S (–60) transgenic plants, respectively. These results showed that the construct of AG-I-35S (–60)::PCS1 was partially effective at preventing seed set and provided a novel sterility strategy. 相似文献
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S. M. Touhidul Islam R. S. Tammi Sneh L. Singla-Pareek Zeba Islam Seraj 《Acta Physiologiae Plantarum》2010,32(4):657-663
Rice yield is severely affected by high-salt concentration in the vicinity of the plant. In an effort to engineer rice for
improved salt tolerance Agrobacterium-mediated transformation of rice cv. Binnatoa was accomplished with the Pennisetum glaucum vacuolar Na+/H+ antiporter gene (PgNHX1) under the constitutive CaMV35S promoter. For the molecular analysis of putative transgenic plants, PCR and RT-PCR were performed.
Transgenic rice plants expressing PgNHX1 showed better physiological status and completed their life cycle by setting flowers and seeds in salt stress, while wild-type
plants exhibited rapid chlorosis and growth inhibition. Moreover, transgenic rice plants produced higher grain yields than
wild-type plants under salt stress. Assessment of the salinity tolerance of the transgenic plants at seedling and reproductive
stages demonstrated the potential of PgNHX1 for imparting enhanced salt tolerance capabilities and improved yield. 相似文献
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Phytochelatins (PCs) are post-translationally synthesized thiol reactive peptides that play important roles in detoxification
of heavy metal and metalloids in plants and other living organisms. The overall goal of this study is to develop transgenic
plants with increased tolerance for and accumulation of heavy metals and metalloids from soil by expressing an Arabidopsis
thaliana
AtPCS1 gene, encoding phytochelatin synthase (PCS), in Indian mustard (Brassica juncea L.). A FLAG-tagged AtPCS1 gDNA, under its native promoter, is expressed in Indian mustard, and transgenic pcs lines have been compared with wild-type
plants for tolerance to and accumulation of cadmium (Cd) and arsenic (As). Compared to wild type plants, transgenic plants
exhibit significantly higher tolerance to Cd and As. Shoots of Cd-treated pcs plants have significantly higher concentrations
of PCs and thiols than those of wild-type plants. Shoots of wild-type plants accumulated significantly more Cd than those
of transgenic plants, while accumulation of As in transgenic plants was similar to that in wild type plants. Although phytochelatin
synthase improves the ability of Indian mustard to tolerate higher levels of the heavy metal Cd and the metalloid As, it does
not increase the accumulation potential of these metals in the above ground tissues of Indian mustard plants. 相似文献
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Citrus FT (CiFT) cDNA, which promoted the transition from the vegetative to the reproductive phase in Arabidopsis thaliana, when constitutively expressed was introduced into trifoliate orange (Poncirus trifoliata L. Raf.). The transgenic plants in which CiFT was expressed constitutively showed early flowering, fruiting, and characteristic morphological changes. They started to
flower as early as 12 weeks after transfer to a greenhouse, whereas wild-type plants usually have a long juvenile period of
several years. Most of the transgenic flowers developed on leafy inflorescences, apparently in place of thorns; however, wild-type
adult trifoliate orange usually develops solitary flowers in the axils of leaves. All of the transgenic lines accumulated
CiFT mRNA in their shoots, but there were variations in the accumulation level. The transgenic lines showed variation in phenotypes,
such as time to first flowering and tree shape. In F1 progeny obtained by crossing ‘Kiyomi’ tangor (C. unshiu × sinensis) with the pollen of one transgenic line, extremely early flowering immediately after germination was observed. The transgene
segregated in F1 progeny in a Mendelian fashion, with complete co-segregation of the transgene and the early flowering phenotype. These results
showed that constitutive expression of CiFT can reduce the generation time in trifoliate orange. 相似文献
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We have established a shoot regeneration system and genetic transformation of cockscomb (Celosia cristata and Celosia plumosus). The best results in terms of frequency of shoot regeneration and number of shoot buds per explant are observed on media
supplemented with 0.5 mg l−1 6-BA (for explants of apical meristems of C. cristata) or 2.0 mg l−1 6-BA, 0.5 mg l−1 NAA and 0.5 mg l−1 IAA (for hypocotyls explants of C. plumosus). We use apical meristems of C. cristata and hypocotyls of C. plumosus as the starting material for transformation. A novel KNOTTED1-like homeobox1 (KNOX), PttKN1 (Populus tremula × P. tremuoides
knotted1) isolated from the vascular cambial region of hybrid aspen, is introduced into cockscomb by Agrobacterium. A series of novel phenotypes are obtained from the transgenic cockscomb plants, including lobed or rumpled leaves, partite
leaves and two or three leaves developed on the same petiole, on the basis of their leaf phenotypes. Transformants are selected
by different concentrations of kanamycin. Transformants are confirmed by PCR of the NptII gene and PCR or RT-PCR of PttKN1 gene. Furthermore, RT-PCR shows that 35S:: PttKN1 RNA levels do not correlate with phenotypic severity. It is discussed that our results bring elements on possible function
of PttKN1 gene. To our knowledge, genetic transformation of cockscomb is first reported. 相似文献
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Guiping Diao Yucheng Wang Chao Wang Chuanping Yang 《Plant Molecular Biology Reporter》2011,29(1):77-87
Plant glutathione S-transferases (GSTs) are involved in protecting plants against both diverse biotic and abiotic stresses. In the present study,
a novel GST gene (LbGST1) was cloned from Limonium bicolor (Bunge) Kuntze (Plumbaginaceae). To characterize its function in salt tolerance, tobacco lines transformed with LbGST1 were generated. Compared with wild-type (WT) tobacco, transgenic plants overexpressing LbGST1 exhibited both GST and glutathione peroxidase activities. Moreover, superoxide dismutase, peroxidase (POD), and catalase
activities in transgenic plants were significantly higher than those in WT plants, particularly when grown under conditions
of salt stress. Similarly, levels of proline in transgenic plants were also higher than those in WT plants grown under NaCl
stress conditions. Whereas, Malondialdehyde contents in transgenic plants were lower than those in WT plants under NaCl conditions.
Furthermore, Na+ content in transgenic plants was lower than that in WT plants under these stress conditions. Subcellular localization analysis
revealed that the LbGST1 protein was localized in the nucleus. These results suggested that overexpression of LbGST1 gene can affect many physiological processes associated with plant salt tolerance. Therefore, we hypothesize that LbGST1 gene can mediate many physiological pathways that enhance stress resistance in plants. 相似文献
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F. D. Espasandin M. M. Collavino C. V. Luna R. C. Paz J. R. Tarragó O. A. Ruiz L. A. Mroginski P. A. Sansberro 《Plant Cell, Tissue and Organ Culture》2010,102(2):181-189
A protocol for the production of transgenic plants was developed for Lotus tenuis via Agrobacterium-mediated transformation of leaf segments. The explants were co-cultivated (for 3 days) with an A. tumefaciens strain harbouring either the binary vector pBi RD29A:oat arginine decarboxylase (ADC) or pBi RD29A:glucuronidase (GUS), which
carries the neomycin phosphotransferase II (nptII) gene in the T-DNA region. Following co-cultivation, the explants were cultured in Murashige and Skoog medium supplemented
with naphthalenacetic acid (NAA) and benzyladenine (BA) and containing kanamycin (30 μg ml−1) and cefotaxime (400 μg ml−1) for 45 days. The explants were subcultured several times (at 2-week intervals) to maintain the selection pressure during
the entire period. About 40% of the explants inoculated with the pBiRD29:ADC strain produced eight to ten adventitious shoots
per responsive explant through a direct system of regeneration, whereas 69% of the explants inoculated with the pBi RD29A:GUS
strain produced 13–15 adventitious shoots per responsive explant. The selected transgenic lines were identified by PCR and
Southern blot analysis. Three ADC transgenic lines were obtained from 30 infected explants, whereas 29 GUS transgenic lines
were obtained from 160 explants, corresponding to a transformation efficiency of 10 and 18.1%, respectively. More than 90%
of the in vitro plantlets were successfully transferred to the soil. The increase in the activity of arginine decarboxylase
from stressed ADC- Lt19 lines was accompanied by a significant rise in the putrescine level. The GUS transgenic line driven by the RD29A promoter
showed strong signals of osmotic stress in the leaves and stem tissues. All of the transgenic plants obtained exhibited the
same phenotype as the untransformed controls under non-stress conditions, and the stability of the gene introduced into the
cloned materials was established. 相似文献
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Hongbo Liu Xiang Guo Muhammad S. Naeem Dan Liu Ling Xu Wenfang Zhang Guixiang Tang Weijun Zhou 《Plant Cell, Tissue and Organ Culture》2011,106(1):143-151
Plant diseases and insect pests are serious threat to the growth and yield of oilseed rape. In this study, a binary vector
carrying sporamin and chitinase
PjChi-1 genes in tandem was introduced into Brassica napus cv. ZS 758 via Agrobacterium tumefaciens for dual resistance against disease and insect attack. Thirty-two regenerated plantlets exhibiting hygromycin resistance
were selected following Agrobacterium-mediated transformation of 600 leaf petiole explants. Of these, 27 transformants were confirmed to carry the two transgenes
as detected by polymerase chain reaction (PCR) with 4.5% transformation efficiency. Eight plantlets were randomly selected
for further confirmation by Southern and northern blot hybridization analyses. Four plants carried single copy of the transgenes,
while the remaining four plants carried either two or three copies of the transgenes. Moreover, expression of the sporamin transgene was detected by northern blot hybridization in transgenic lines, but not in wild-type plants. These eight T0 plants were grown in vitro, and inoculated with the Lepidoptera larvae of Plutella xylostella and with spores of the fungal pathogen of Sclerotinia sclerotiorum. Transgenic plants exhibited high levels of resistance to P. xylostella and S. sclerotiorum when compared to untransformed wild-type plants. Genetic analysis of T1 progeny confirmed Mendelian segregation of the introduced genes. Therefore, these transgenic lines demonstrate a promising
potential for variety development of oilseed rape lines with enhanced resistance against both P. xylostella and S. sclerotiorum. 相似文献
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The Nicotiana tabacum transgenic plants expressing a Cucurbita pepo antisense PHYA RNA were obtained. The seedlings of transgenic tobacco with reduced phytochrome A (PHYA) content displayed decreased sensitivity
to continuous broad-band far-red radiation (λ > 680 nm). Under far-red irradiance transgenic seedlings showed less elongation
of the hypocotyls, more rapid plastid development, more chlorophyll accumulation, less repression of lightdependent NADPH:protochlorophyllide
oxidoreductase than wild-type plants that was in accordance with PHYA control of plant development. Dynamics of the far-red
radiation dependent changes in low temperature chlorophyll fluorescence spectra for the transgenic and wild-type seedlings
were consistent with the more rapid formation of photosynthetic apparatus in the seedlings with reduced PHYA. 相似文献
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Olga A. Shulga Tatyana Yu Mitiouchkina Anna V. Shchennikova Konstantin G. Skryabin Sergey V. Dolgov 《In vitro cellular & developmental biology. Plant》2011,47(5):553-560
Chrysanthemum is one of the most important commercial cut flowers in the world. Early-flowering cultivars are required to
produce quality chrysanthemum flowers with a lower cost of production. To shorten the vegetative growth phase of chrysanthemum,
three AP1-like genes from Asteraceae were constitutively overexpressed in 80 independent transgenic chrysanthemum lines. All lines were characterized by PCR and
RT-PCR and demonstrated that overexpression of compositae AP1-homologs in transgenic chrysanthemum under long-day conditions had no effect on plant development compared to non-transgenic
controls. Conversely, under short-day conditions, transgenic plants commenced bud initiation 2 wk earlier than non-transgenic
chrysanthemum plants. Subsequently, transgenic chrysanthemum flowers showed color earlier and resulted in full opening of
inflorescences 3 wk prior to non-transgenic control plants. These results open new possibilities for genetic improvement and
breeding of chrysanthemum cultivars. 相似文献
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The suadea salsa full-length S-adenosylmethionine synthetase (SsSAMS2) was introduced into tobacco (Nicotiana tabacum L.) by Agrobacterium tumefaciens-mediated transformation. The gene transformation and expression in tobacco were confirmed by PCR, RT-PCR and Northern blotting
analysis. Several transgenic lines (ST lines) overexpressing SsSAMS2 gene under the control of cauliflower mosaic virus 35S promoter showed more seeds number and weight, and accumulated higher
free total polyamines (PAs) than wild-type plants (WT lines) and transformants with blank vector (BT lines). Salt stress-induced
damage was attenuated in these transgenic plants, in the symptom of maintaining higher photosynthetic rate and biomass. These
results that the transgenic plants overexpressing suadea salsa SAMS2 are more tolerant to salt stress than wild-type plants suggest that PAs may play an important role in contributing salt tolerance
to plants. 相似文献
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Plant aquaporins are believed to facilitate water transport across cell membranes. However, the relationship between aquaporins
and drought resistance in plants remains unclear. VfPIP1, a putative aquaporin gene, was isolated from Vicia faba leaf epidermis, and its expression was induced by abscisic acid (ABA). Our results indicated that the VfPIP1 protein was
localized in the plasma membrane, and its expression in V. faba was induced by 20% polyethylene glycol 6000. To further understand the function of VfPIP1, we obtained VfPIP1-expressing transgenic Arabidopsis thaliana plants under the control of the CaMV35S promoter. As compared to the wild-type control plants, the transgenic plants exhibited
a faster growth rate, a lower transpiration rate, and greater drought tolerance. In addition, the stomata of the transgenic
plants closed significantly faster than those of the control plants under ABA or dark treatment. These results suggest that
VfPIP1 expression may improve drought resistance of the transgenic plants by promoting stomatal closure under drought stress. 相似文献
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Ok-Sun?Lee Young-Min?Kang Hee-Young?Jung Ji-Yun?Min Seung-Mi?Kang Chandrakant?S.?Karigar D.?Theertha?Prasad Jung-Dong?Bahk Myung-Suk?Choi
Summary In wild-type Scopolia parvilfora (Solanaceae) tissues, only the roots express the enzyme putrescine N-methyltransferase (PMT; EC 2.1.1.53), which is the first specific precursor of the tropane alkaloids. Moreover, the tropanane
alkaloid levels were the highest in the root (0.9 mg g−1 on a dry weight basis), followed by the stem and then the leaves. We metabolically engineered S. parviflora by introducing the tobacco pmt gene into its genome by a binary vector system that employs disarmed Agrobacterium rhizogenes. The kanamycin-resistant hairy root lines were shown to bear the pmt gene and to overexpress its mRNA and protein product by at least two-fold, as determined by polymerase chain reaction (PCR)
and Northern and Western blottings, respectively. The transgenic lines also showed higher PMT activity and were morphologically
aberrant in terms of slower growth and the production of lateral roots. The overexpression of pmt markedly elevated the scopolamine and hyoscyamine levels in the transgenic lines that showed the highest pmt mRNA and PMT protein levels. Thus, overexpression of the upstream regulator of the tropane alkaloid pathway enhanced the
biosynthesis of the final product. These observations may be useful in establishing root culture systems that generate large
yields of tropane alkaloids.
These authors contributed equally to this paper (co-first authors). 相似文献
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Y. Q. Zhou H. Y. Duan C. E. Zhou J. J. Li F. P. Gu F. Wang Z. Y. Zhang Z. M. Gao 《Russian Journal of Plant Physiology》2009,56(2):224-231
The objective of this research was to establish an efficient system of genetic transformation and plant regeneration from
hairy roots by infecting the leaf sections and stem segments of in vitro Rehmannia glutinosa Libosch. f. hueichingensis Hsiao plantlets. Hairy roots were induced from them after co-culturing with Agrobacterium rhizogenes strain 15834 at a frequency of 32 and 29.4%, respectively. The calluses were induced from hairy roots on half-strength Murashige
and Skoog medium containing 0.2 mg/l kinetin and 3.0 mg/l benzyladenine at a frequency of 100%, from which transgenic shoots
and plantlets were developed. Transgenic plantlets did not have differences in morphology except the shortened internodes
and an increase in adventitious root formation compared to wild-type plants. PCR and Southern-blot analyses confirmed that
rolB gene of TL-DNA was inserted in the genome of transformed hairy roots and plantlets. RT-PCR analysis and opine paper electrophoresis
revealed that rolB gene was expressed in the transformed hairy roots and plantlets. Conclusively, transgenic hairy roots and transgenic plants
of Rehmannia glutinosa Libosch. f. hueichingensis Hsiao were developed for the first time.
This text was submitted by the authors in English.
Published in Russian in Fiziologiya Rastenii, 2009, Vol. 56, No. 2, pp. 247–255. 相似文献
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Meadow fescue (Festuca pratensis Huds.) is an important cool-season forage grass in Europe and Asia. We developed a protocol for producing meadow fescue transgenic
plants mediated by Agrobacterium tumefaciens transformation. Embryogenic calli derived from mature embryos were transformed with A. tumefaciens strain AGL1 carrying the binary vector pDM805, coding for the phosphinothricin acetyltransferase (bar) and β-glucuronidase (uidA) genes. Bialaphos was used as the selective agent throughout all phases of tissue culture. In total, 40 independent transgenic
plants were recovered from 45 bialaphos-resistant callus lines and an average transformation efficiency of 2% was achieved.
The time frame from infection of embryogenic calli with Agrobacterium to transferring the transgenic plants to the greenhouse was 18 weeks. In a study of 11 BASTA-resistant transgenic lines,
the uidA gene was expressed in 82% of the transgenic lines. Southern blot analysis revealed that 82% of the tested lines integrated
one or two copies of the uidA gene.
C. Gao and J. Liu contributed equally to the work. 相似文献
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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