共查询到20条相似文献,搜索用时 15 毫秒
1.
Transgenic soybean (Glycine max L.) plants derived from electricdischarge particle acceleration experiments exhibited varyingdegrees of chimerism which was followed by the expression ofthe introduced ß-glucuronidase (gus) gene. Degreesof chimerism in transgenic plants were established by determiningexpression of the gus gene observed as blue spots, streaks orsectors in stem and leaf tissues in in vitro grown plantletsand greenhouse plants. Clonal plants were also obtained. Presenceof the gene was confirmed by Southern blot analysis. These studiespermitted the reconstruction of a partial picture for the developmentof the soybean plant. Glycine max L. cv. Williams 82, soybean, transformation, ß-glucuronidase, chimeric plant phenotypes, development 相似文献
2.
I. Fawole 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2001,102(2-3):458-462
A chimeric plant was observed in the F2 generation of a cross between a mutant cultivar, Ife BPC, and a germplasm line, TVu 2, in cowpea, Vigna unguiculata (L.) Walp. The chimeric plant had four lateral branches, one of which was intensely variegated, while the others were mostly
green with few white sectors. F3 progeny from the intensely variegated branch of this plant were all variegated, while seed derived from the mostly green
branches produced only green progeny. In subsequent generations, the descendants of the variegated branch bred true for the
variegated trait, while those of the mostly green branches were also true-breeding for green colour. No pure-green or pure-white
shoots were observed in any of the variegated plants examined in this study. Consequently, no pure-green or pure-white seedlings
were produced from seeds harvested from the variegated plants. The results of reciprocal crosses between variegated and normal
green plants indicate that variegation is inherited in a strictly uniparental maternal fashion. This is the first report of
a cytoplasmically inherited mutation affecting foliage colour in cowpea.
Received: 10 March 2000 / Accepted: 16 May 2000 相似文献
3.
Segregation of transgenes in maize 总被引:23,自引:0,他引:23
T. Michael Spencer James V. O'Brien William G. Start Thomas R. Adams William J. Gordon-Kamm Peggy G. Lemaux 《Plant molecular biology》1992,18(2):201-210
Progeny recovered from backcrossed transgenic maize tissue culture regenerants (R0) were analyzed to determine the segregation, expression, and stability of the introduced genes. Transgenic A188×B73 R0 plants (regenerated from embryogenic suspension culture cells transformed by microprojectile bombardment; see [9]) were pollinated with nontransformed B73 pollen. Inheritance of a selectable marker gene, bar, and a nonselectable marker gene, uidA, was analyzed in progeny (R1) representing four independent transformation events. Activity of the bar gene product, phosphinothricin acetyltransferase (PAT), was assessed in plants comprising the four R1 populations. The number of R1 plants containing PAT activity per total number of R1 plants recovered for each population was 2/7, 19/34, 3/14 and 73/73. Molecular analysis confirmed the segregation of bar in three R1 populations and the lack of segregation in one R1 population. Cosegregation analysis indicated genetic linkage of bar and uidA in all four R1 populations. Analysis of numerous R2 plants derived from crossing transformed R1 plants with nontransformed inbreds revealed 1:1 segregation of PAT activity in three of four lines, including the line that failed to segregate in the R1 generation. Integrated copies of bar in one line appeared to be unstable or poorly transmitted. 相似文献
4.
Ufuk Celikkol Akcay M. Mahmoudian H. Kamci M. Yucel H. A. Oktem 《Plant cell reports》2009,28(3):407-417
A simple and reproducible Agrobacterium-mediated transformation protocol for a recalcitrant legume plant, lentil (Lens culinaris M.) is reported. Application of wounding treatments and efficiencies of three Agrobacterium tumefaciens strains, EHA105, C58C1, and KYRT1 were compared for T-DNA delivery into lentil cotyledonary node tissues. KYRT1 was found
to be on average 2.8-fold more efficient than both EHA105 and C58C1 for producing transient β-glucuronidase (GUS) gene (gus) expression on cotyledonary petioles. Wounding of the explants, use of an optimized transformation protocol with the application
of acetosyringone and vacuum infiltration treatments in addition to the application of a gradually intensifying selection
regime played significant roles in enhancing transformation frequency. Lentil explants were transformed by inoculation with
Agrobacterium tumefaciens strain, KYRT1 harboring a binary vector pTJK136 that carried neomycin phosphotransferase gene (npt-II) and an intron containing gusA gene on its T-DNA region. GUS-positive shoots were micrografted on lentil rootstocks. Transgenic lentil plants were produced
with an overall transformation frequency of 2.3%. The presence of the transgene in the lentil genome was confirmed by GUS
assay, PCR, RT-PCR and Southern hybridization. The transgenic shoots grafted on rootstocks were successfully transferred to
soil and grown to maturity in the greenhouse. GUS activity was detected in vegetative and reproductive organs of T0, T1, T2 and T3 plants. PCR assays of T1, T2 and T3 progenies confirmed the stable transmission of the transgene to the next generations. 相似文献
5.
Effects of overproduction of tobacco MnSOD in maize chloroplasts on foliar tolerance to cold and oxidative stress 总被引:15,自引:1,他引:15
Van Breusegem F; Slooten L; Stassart J; Botterman J; Moens T; Van Montagu M; Inze D 《Journal of experimental botany》1999,50(330):71-78
Transgenic maize plants have been generated by particle gun bombardment
that overproduce a Nicotiana plumbaginifolia L.
manganese superoxide dismutase (MnSOD). To target this mitochondrial enzyme
into chloroplasts, the mature MnSOD-coding sequence was fused to a
chloroplast transit peptide from a pea ribulose-1,5-bisphosphate
carboxylase gene, whereas expression of the chimeric gene was driven by the
CaMB 35S promoter. Transgenic MnSOD activity contributed to 20% of the
total SOD activity. The presence of transgenic MnSOD had clear effects on
foliar tolerance to chilling and oxidative stress. The results suggest that
overproduction of MnSOD in the chloroplasts increases the antioxidant
capacity of the leaves. 相似文献
6.
Finger millet plants conferring resistance to leaf blast disease have been developed by inserting a rice chitinase (chi11) gene through Agrobacterium-mediated transformation. Plasmid pHyg-Chi.11 harbouring the rice chitinase gene under the control of maize ubiquitin promoter
was introduced into finger millet using Agrobacterium strain LBA4404 (pSB1). Transformed plants were selected and regenerated on hygromycin-supplemented medium. Transient expression
of transgene was confirmed by GUS histochemical staining. The incorporation of rice chitinase gene in R0 and R1 progenies was confirmed by PCR and Southern blot analyses. Expression of chitinase gene in finger millet was confirmed by
Western blot analysis with a barley chitinase antibody. A leaf blast assay was also performed by challenging the transgenic
plants with spores of Pyricularia grisea. The frequency of transient expression was 16.3% to 19.3%. Stable frequency was 3.5% to 3.9%. Southern blot analysis confirmed
the integration of 3.1 kb chitinase gene. Western blot analysis detected the presence of 35 kDa chitinase enzyme. Chitinase
activity ranged from 19.4 to 24.8. In segregation analysis, the transgenic R1 lines produced three resistant and one sensitive for hygromycin, confirming the normal Mendelian pattern of transgene segregation.
Transgenic plants showed high level of resistance to leaf blast disease compared to control plants. This is the first study
reporting the introduction of rice chitinase gene into finger millet for leaf blast resistance. 相似文献
7.
Werner Kamberger 《Archives of microbiology》1979,121(1):83-90
Acidic exopolysaccharides and O-antigen containing lipopolysaccharides were isolated from Rhizobium japonicum, R. leguminosarum, R. lupini, R. meliloti, R. phaseoli, cowpea Rhizobium sp. and a non-nodulating soil bacterium. Lectins from seeds of soybean (Glycine max), garden pea (Pisum sativum), lentil (Lens culinaris), alfalfa (Medicago sativa), field bean (Phaseolus vulgaris), jackbean (Canavalia ensiformis) and from wheat germ were tested for their capacity to precipitate rhizobial exopolysaccharides and lipopolysaccharides in the Ouchterlony double diffusion test. Soybean lectin precipitated exclusively with the exopolysaccharide of R. japonicum, whereas the lectins from pea and lentil precipitated exopolysaccharides from all the fast growing strains of Rhizobium. Host range specific interactions between lipopolysaccharides and lectins were observed in the pea/lentil-R. leguminosarum and in the alfalfa-R. meliloti systems. Concanavalin A precipitated the exopolysaccharides of all fast growing strains of Rhizobium, the exopolysaccharide of the cowpea strain and several lipopolysaccharides of different Rhizobium species and thus did not show any correlation between polysaccharide binding and symbiotic specificity. Non-leguminous wheat germ agglutinin did not precipitate any of the rhizobial polysaccharides tested and the lipopolysaccharide of the soil bacterium did not precipitate with any of the lectins examined.Abbreviations Con A
Concanavalin A
- CPC
cetylpyridinium chioride
- EPS
exopolysaccharide
- FITC
fluorescein isothiocyanate
- KDO
2-keto-3-deoxyoctonic acid
- LPS
lipopolysaccharide
- PBS
phosphate-buffered saline
- PS
polysaccharide 相似文献
8.
A new plant expression vector (pBSbtCry1Ac-GNA) containing two insect resistant genes, a synthetic chimeric gene SbtCry1Ac encoding the insecticidal protein CrylAc and a gene GNA encoding snowdrop lectin (Galanthus nivalis agglutinin) was constructed. Transgenic tobacco plants containing these two genes were obtained through Agrobacterium-mediated transformation of tobacco leaf discs. Results from PCR detection and genomic DNA Southern blot analysis indicated
that both SbtCrylAc gene and GNA gene were integrated into the genome of these plants. Results of Western blot analysis indicated that these two proteins
were expressed in the analyzed plants. Bioassays of Myzus persicae and Helicoverpa assulta on detached leaves of transformed tobacco plants were carried out. The average aphid inhibition rate of these plants tested
at 12 d post-infestation was 71.9 %. The average H. assulta mortality of these plants tested at 6 d post-infestation was up to 89.8 %. The kanamycin resistance of the T1 progeny of these transgenic plants was analyzed and a typical 3:1 segregation was observed. 相似文献
9.
Novel insect resistance in Brassica napus developed by transformation of chitinase and scorpion toxin genes 总被引:5,自引:0,他引:5
Transgenic plants with introduced pest-resistant gene offer an efficient alternative insect control. The novel insect-resistant gene combination, chitinase(chi) and BmkIT(Bmk), containing an insect-specific chitinase gene and a scorpion insect toxin gene was introduced into Brassica napus cultivar via Agrobacterium-mediated transformation. Fifty-seven regenerated plantlets with kanamycin-resistance were obtained. Transgenic plants were verified by Southern blot analysis. Enzyme-linked immunosorbent assay (ELISA) and bioassay of artificial inoculation with diamondback moth (Plutella maculipenis) (DBM) larvae indicated that some of the transgenic plants were high-level expression for both chitinase and scorpion toxin proteins and performed high resistance against the tested pest infestation. The genetic analysis of T1 progeny confirmed that the inheritance of introduced genes followed the Mendelian rules. 相似文献
10.
Kitisri Sukhapinda Rosa Spivey Robert B. Simpson Elias A. Shahin 《Molecular & general genetics : MGG》1987,206(3):491-497
Summary We transformed tomato (Lycopersicon esculentum L.) by using Agrobacterium rhizogenes containing two independent plasmids: the wild-type Ri-plasmid, and the vector plasmid, pARC8. The T-DNA of the vector plasmid contained a marker gene (Nos/Kan) encoding neomycin phosphotransferase which conferred resistance to kanamycin in transformed plant cells. Transgenic plants (R
0) with normal phenotype were regenerated from transformed organogenic calli by the punctured cotyledon transformation method. Southern blot analysis of the DNA from these transgenic plants showed that one or two copies of the vector plasmid T-DNA, but none of the Ri-plamid T-DNA, were integrated into the plant genome. Different transgenic plants derived from the same callus clone showed an identical DNA banding pattern, indicating the non-chimeric origin of these plants. We also transformed tomato by using A. tumefaciens strain LBA4404 containing a disarmed Ti-plasmid (pAL4404), and a vector plasmid (pARC8). Transgenic plants derived via A. tumefaciens transformation, like those via A. rhizogenes, contained one to two copies of the integrated vector T-DNA. The kanamycin resistance trait in the progeny (R
1) of most transgenic plants segregated at a ratio of 3:1, suggesting that the vector T-DNAs were integrated at a single site on a tomato chromosome. In some cases, the expression of the marker gene (Nos/Kan) seemed to be suppressed or lost in the progeny. 相似文献
11.
Transformation of Soybean (Glycine max) by Infecting Germinating Seeds with Agrobacterium tumefaciens 总被引:9,自引:1,他引:8
下载免费PDF全文
![点击此处可从《Plant physiology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
The transfer of genetic material into soybean tissue was accomplished by using an avirulent strain of Agrobacterium tumefaciens which contained the binary vector pGA482. The method used for transformation requires no tissue culture steps as it involves the inoculation of the plumule, cotyledonary node, and adjacent cotyledon tissues of germinating seeds. The identification of neomycin phosphotransferase (NPT) II enzyme activity in the tissues of 16 (R0) soybean plants indicated that the plant expressible Nos-NPT II gene, contained within the T-DNA region from pGA482, had been transferred at least into somatic tissues. Putative transformed R0 soybean plants were advanced to produce R1 plants which were also assayed for the presence of the transferred Nos-NPT II gene. The combined results of these assays indicated that about 0.7% of the surviving inoculated seeds yielded transformed tissues in the R0 plant, and that about 1/10 of these plants yielded transformed R1 plants. The presence of the Nos-NPT II gene in DNAs isolated from both R0 and R1 plant was demonstrated by using genomic blot hybridization and polymerase chain reaction methods. Integration of this gene into the soybean genome was demonstrated for three R1 soybean plants. 相似文献
12.
Zhang J. Van Toai T. Huynh L. Preiszner J. 《Molecular breeding : new strategies in plant improvement》2000,6(2):135-144
Flooding is one of the most serious environmental stresses that affect plant growth and productivity. Flooding causes premature senescence which results in leaf chlorosis, necrosis, defoliation, cessation of growth and reduced yield. This study was conducted to determine the effects of autoregulated cytokinin production on the flooding tolerance of Arabidopsis thaliana plants. A chimeric gene containing the senescence-specific SAG12 promoter and the ipt gene coding for isopentenyl transferase, a rate-limiting enzyme in the cytokinin biosynthesis pathway, was constructed. The chimeric gene was introduced into Arabidopsis plants by Agrobacterium-mediated vacuum infiltration. Four transgenic lines were chosen for flooding tolerance determinations. DNA hybridization analysis and PCR confirmed that all four of the transgenic lines carried the ipt gene. The segregation of kanamycin resistance in the T2 generation indicated 1 to 3 integration events. GUS expression and RT-PCR of the ipt gene confirmed the senescence-specificity of the SAG12 promoter. Morphologically, the transgenic lines appeared healthy and normal. Transgenic plants began to flower at the same time as wild-type plants, but the period from flowering to senescence was lengthened by 7 to 12 days. Tolerance of the transgenic plants to waterlogging and complete submergence was assayed in three independent experiments. All four transgenic lines were consistently more tolerant to flooding than wild-type plants. The results indicated that endogenously produced cytokinin can regulate senescence caused by flooding stress, thereby, increasing plant tolerance to flooding. This study provides a novel mechanism to improve flooding tolerance in plants. 相似文献
13.
Kalbina I Wallin A Lindh I Engström P Andersson S Strid K 《Protein expression and purification》2011,80(2):194-202
The major outer membrane protein (MOMP) of Chlamydia trachomatis is a highly antigenic and hydrophobic transmembrane protein. Our attempts to express the full-length protein in a soluble form in Escherichia coli and in transgenic plants failed. A chimeric gene construct of C. trachomatis serovar E MOMP was designed in order to increase solubility of the MOMP protein but with retained antigenicity. The designed construct was successfully expressed in E. coli, in Arabidopsis thaliana, and in Daucus carota. The chimeric MOMP expressed in and purified from E. coli was used as antigen for production of antibodies in rabbits. The anti-chimeric MOMP antibodies recognized the corresponding protein in both E. coli and in transgenic plants, as well as in inactivated C. trachomatis elementary bodies. Transgenic Arabidopsis and carrots were characterized for the number of MOMP chimeric genetic inserts and for protein expression. Stable integration of the transgene and the corresponding protein expression were demonstrated in Arabidopsis plants over at least six generations. Transgenic carrots showed a high level of expression of the chimeric MOMP – up to 3% of TSP. 相似文献
14.
A. Chaparro-Giraldo R. M. Barata S. M. Chabregas R. A. Azevedo M. C. Silva-Filho 《Plant cell reports》2000,19(10):961-965
Potato tubers were transformed with a chimeric gene made by the fusion of the soybean leghemoglobin encoding gene (lba) with the chloroplastic targeting sequence from Rubisco. This construct was placed under the control of the strong constitutive
35S promoter and the 3′ nontranslated region of Rubisco from pea. Leghemoglobin expression on kanamycin-resistant plants was
monitored by RT-PCR. Furthermore, immunodetection of subcellular fractions of transgenic plants revealed that leghemoglobin
was imported and correctively processed inside the organelle. In addition, analysis of transgenic plants revealed reduced
growth and decreased tuber production compared with the untransformed plants. It is suggested that leghemoglobin expression
in potato chloroplasts interferes with aerobic metabolism, leading to physiological and morphological changes.
Received: 20 December 1999 / Revision received: 28 May 2000 / Accepted: 16 June 2000 相似文献
15.
Gurdip S. Brar Barry A. Cohen Carole L. Vick Grant W. Johnson 《The Plant journal : for cell and molecular biology》1994,5(5):745-753
Transgenic plants of Florunner and Florigiant, two of the most widely cultivated peanut cultivars in the USA, have been developed using the ACCELL® gene delivery method. Shoot meristems of mature embryonic axes were bombarded with gold beads coated with DNA encoding β-glucuronidase (gus), phosphinothricin acetyl transferase (bar), and tomato spotted wilt virus-nucleocapsid protein (tswv-np) genes. Transgenic shoots were identified by screening for GUS activity, and independent transformants were recovered from both cultivars. Molecular analysis of two of these transformants in R0 and R1 generations demonstrated the stable integration of the foreign genes into the plant genome. One transgenic plant had one to two copies of the genes integrated into the genome of its progeny, whereas the other had multiple copies. Gus and bar genes exhibited predictable segregation ratios in the R1 and R2 generations and were genetically linked. Integration of the bar gene conferred resistance to BASTATM, a wide-spectrum herbicide, applied at 500 p.p.m. of active ingredient. Resistance of the transgenic plants to tomato spotted wilt virus is currently being tested under greenhouse conditions. The ACCELL® particle bombardment system is expected to be effective for transformation of a wide variety of commercial peanut cultivars. 相似文献
16.
Generation of reactive oxygen species (ROS) and activities of antioxidant enzymes (catalase, peroxidase, ascorbate peroxidase)
in pea (Pisum sativum L.) and soybean (Glycine max L.) under hypoxia (3–24 h) and high CO2 concentration in medium were studied. In sensitive to hypoxia pea seedlings, hypoxia enhanced markedly production of superoxide
anion-radical, hydroperoxides, and especially hydrogen peroxide. In more tolerant soybean plants, these changes were less
pronounced. During first hours of hypoxia, activity of lipoxygenase in plant cells increased. This allows a suggestion that
this enzyme is involved in the processes of hydroperoxide accumulation in plant tissues under oxygen deficit. In pea and soybean
plants, a correlation between tolerance to hypoxia, the rate of ROS generation, and antioxidant enzyme activities was established.
During the first hours of hypoxia, the catalase activity in soybean plants increased stronger than in sensitive to hypoxia
pea plants. At longer exposure to hypoxia (24 h), peroxidases started to play the higher role in cell defense against hypoxia,
but only in soybean plants. The medium with the higher CO2 content induced higher changes in the processes of ROS accumulation and activities of lipoxygenase and antioxidant enzymes.
This permits us to refer CO2, accumulated as a product of respiration in the cells, to low-molecular signal molecules switching on plant adaptation to
hypoxic stress. 相似文献
17.
Uchimiya Hirofumi Fujii Seiichi Huang Jirong Fushimi Takaomi Nishioka Masanori Kim Kyung-Min Yamada Maki Kawai Kurusu Takamitsu Kuchitsu Kazuyuki Tagawa Michito 《Molecular breeding : new strategies in plant improvement》2002,9(1):25-31
We isolated a rice gene (denoted YK1), which showed78 percent amino acid sequence homology to the maize HM1gene. A chimeric gene consisting of a promoter and first intron of maizeubiquitin gene and the cDNA of YK1 was introduced intorice via Agrobacterium mediated transformation. Transgenic riceplants overexpressing this chimeric gene were resistant to rice blast(Magnaporthe grisea) disease, which is one of the mostserious pathogens in rice. Furthermore, the same transgenic plants conferredhigh tolerance to several abiotic stresses such as NaCl, UV-C, submergence, andhydrogen peroxide. 相似文献
18.
Prediction of germ-line transformation events in chimeric Ro transgenic soybean plantlets using tissue-specific expression patterns 总被引:1,自引:1,他引:0
Paul Christou Dennis E. McCabe 《The Plant journal : for cell and molecular biology》1992,2(3):283-290
Chimeric soybean plants derived from electric discharge particle acceleration experiments were used to develop relationships between tissue-specific expression patterns of a marker gene and germ-line transformation events in Ro plants. Using the GUS histochemical assay the likelihood of germ-line transformation was correlated to the localization of enzyme activity in specific tissues of the parent plant. The expression patterns of the 35S-GUS marker gene in specific tissue types in Ro plants and correlation with transmission of the introduced gene to progeny allowed the production of a clear picture of the development of these plants and their germ-line cells. Advances reported in this article resulted in the development of a commercial process for the genetic improvement of this important staple crop which until recently was very recalcitrant to conventional transformation methods. 相似文献
19.
Summary Fifteen isolates of nodule bacteria were isolated from root and stem nodules ofAeschynomene aspera and they were characterized as Rhizobium by well known laboratory tests. All these isolates together with other efficient strains of known rhizobia belonging to different cross-inoculation groups were evaluated for their nodulation abilities onAeschynomene aspera, Cajanus cajan (pigeon pea),Cicer arietinum (chickpea),Pisum sativum (pea),Trifolium repens (clover),Medicago sativa (lucerne),Lens culinaris (lentil),Glycine max (soybean),Vigna sinensis (cowpea),Vigna radiata (mung bean),Vigna mungo (urd bean) andArachis hypogea (peanut). The results demonstrated that Rhizobium fromAeschynomene could form nodules only on its homologous host (Aeschynomene) but not on other legumes tested. Secondly, none of the rhizobia of other cross-inoculation groups could nodulateA. aspera. 相似文献
20.
S. V. Ramu S. Rohini G. Keshavareddy M. Gowri Neelima N. B. Shanmugam A. R. V. Kumar S. K. Sarangi P. Ananda Kumar M. Udayakumar 《Journal of Applied Entomology》2012,136(9):675-687
Pigeon pea is an important legume. Yield losses due to insect pests are enormous in the cultivation of this crop. Expression of cry proteins has led to increased resistance to pests in several crops. We report in this paper, expression of a chimeric cry1AcF (encoding cry1Ac and cry1F domains) gene in transgenic pigeon pea and its resistance towards Helicoverpa armigera. PCR, Southern hybridization, RT‐PCR and Western analysis confirmed stable integration and expression of the cry1AcF gene in pigeon pea transgenics. When screened for efficacy of the transformants for resistance against H. armigera, the transgenics showed not only high mortality of the larva but could also resist the damage caused by the larvae. Analysis for the stable integration, expression and efficacy of the transgenics resulted in the identification of four T3 plants arising from two T1 backgrounds as highly promising. The results demonstrate potentiality of the chimeric cry1AcF gene in developing H. armigera‐resistant pigeon pea. 相似文献