• Title/Summary/Keyword: Plant transformation

Search Result 816, Processing Time 0.027 seconds

Agrobacterium-mediated transformation of Lycopersicon esculentum (cv. MicroTom) with two pathogen-induced hot pepper transcription factors

  • Seong, Eun-Soo;Oh, Sang-Keun;Eunsook Chung;Park, Doil
    • Proceedings of the Korean Society of Plant Pathology Conference
    • /
    • 2003.10a
    • /
    • pp.79.1-79
    • /
    • 2003
  • Two pathogen-induced hot pepper transcription factors (CaNACl and CapIfl) were introduced into‘MicroTom’tomato by Agrobacterium tumefaciens-mediated transformation. We used to nptII containing kanamycin resistance gene as a selection marker. Both transformed and non-transformed plants were transferred to pot after rooting test in vitro. To approximate the levels of caNACl transcript in leaves of wild-type and transgenic plants, RNA blots were hybridized with double-stranded full-length CaNACl probe at moderate stringency, Although the relative signal strength for hybridization fluctuated among the samples on different blots, transgenic plant lines N-1, N-2 and N-3 consistently displayed increased levels of CaNACl transcript relative to other transgenic lines and wild-type plants. Of all the transgenic lines examined, line N-7 had the least amount of CaNACl transcript. Role of these transcription factors in pathogen defense will be examined by overexpression in tomato.

  • PDF

Biolistic transformation of Moroccan durum wheat varieties by using mature embryo-derived calli

  • Senhaji, Chaimae;Gaboun, Fatima;Abdelwahd, Rabha;Diria, Ghizlane;Udupa, Sripada;Douira, Allal;Iraqi, Driss
    • Journal of Plant Biotechnology
    • /
    • v.48 no.4
    • /
    • pp.246-254
    • /
    • 2021
  • Environmental stresses are estimated to have reduced global crop yields of wheat by 5.5%. However, traditional approaches for the transfer of resistance to these stresses in wheat plants have yielded limited results. In this regard, genetic transformation has undoubtedly opened up new avenues to overcome crop losses due to various abiotic stresses. Particle bombardment has been successfully employed for obtaining transgenic wheat. However, most of these procedures employ immature embryos, which are not available throughout the year. Therefore, the present investigation utilized mature seeds as the starting material and used the calli raised from three Moroccan durum wheat varieties as the target tissue for genetic transformation by the biolistic approach. The pANIC-5E plasmid containing the SINA gene for drought and salinity tolerance was used for genetic transformation. To enhance the regeneration capacity and transformation efficiency of the tested genotypes, the study compared the effect of copper supplementation in the induction medium (up to 5 μM) with the standard MS medium. The results show that the genotypes displayed different sensitivities to CuSO4, indicating that the transformation efficiency was highly genotype-dependent. The integration of transgenes in the T0 transformants was demonstrated by polymerase chain reaction (PCR) analysis of the obtained resistant plantlets with primers specific to the SINA gene. Among the three genotypes studied, 'Isly' showed the highest efficiency of 9.75%, followed by 'Amria' with 1.25% and 'Chaoui' with 1%.

Construction of Binary Vectors for the Rice Transformation Using a Rice Actin Promoter and Replication Origin of pTi12 Isolated from Agrobacterium tumefaciens KU12 (Agrobactrium tumefaciens KU12로부터 분리한 pli12의 Replication Origin과 벼의 Actin 유전자 프로모터를 이용한 벼의 Binary Vector 제조)

  • Sim, Woong-Seop
    • Journal of Plant Biology
    • /
    • v.38 no.4
    • /
    • pp.365-371
    • /
    • 1995
  • Binary vectors, pBI-ActR1, pBI-ActF1 and pBSH-ActR1, were constructed using pGA642, the replication origin of pTi12 and the rice actin promoter. The sizes of pBI-ActR1, pBI-ActF1 and pBSH-ActR1 were 12.9 kb, 13.2 kb and 11.95 kb, respectively. These vectors containing a rice actin promoter followed by a GUS structural gene could induce stronly the expression of GUS gene in transformed rice cells. Rice explants from 3-4 day old seedlings after germinatin were cocultured with A. tumefaceins harboring pBI-ActR1, pBI-ActF1 or pBSH-ActR1, and then GUS expression in the explants was assayed. Transformation of rice explants by these binary vectors was tissue-specific, such that the meristematic regions of shoot apex, root and hypocotyl were transformed by these binary vectors.

  • PDF

Chloroplast Genetic Transformation in Higher Plants: An Encounter Between Prokaryote and Eukaryote (고등식물의 엽록체 형질전환: 원핵생물과 진맥생물의 조우)

  • Chung, Hwa-Jee;Suh, Young-Bae;Jeong, Won-Joong;Min, Sung-Ran;Liu, Jang-R.
    • Journal of Plant Biotechnology
    • /
    • v.33 no.3
    • /
    • pp.185-194
    • /
    • 2006
  • Chloroplasts are believed to be descended from certain cyanobacteria, which were taken up by phagocytosis into a host cell and lived there in a symbiotic relationship. In contrast to the current static concept on the chloroplast genome, its dynamism has been recently demonstrated: the chloroplast genome is active in intramolecular homolgous recombination, producing subgenomic circles when it obtains homolgous sequences via genetic transformation. Chloroplast tranformation in higher plants provides many advantages over nuclear transformation that include higher expression levels of transgenes, polycistronic expression of transgenes, and maternal transmission of transgenes. Tobacco has been used as a model for chloroplast genetic transformation. However, it is recently possible to transform the chloroplasts of other major food and economic crops including rice, soybean, and cotton. Chloroplast-transformed crops will be able to replace bioreactors using microorganisms for production of value-added proteins in future.

Efficient transformation of Actinidia arguta by reducing the strength of basal salts in the medium to alleviate callus browning

  • Han, Meili;Gleave, Andrew P.;Wang, Tianchi
    • Plant Biotechnology Reports
    • /
    • v.4 no.2
    • /
    • pp.129-138
    • /
    • 2010
  • An efficient transformation system for high-throughput functional genomic studies of kiwifruit has been developed to overcome the problem of necrosis in Actinidia arguta explants. The system uses Agrobacterium tumefaciens strain EHA105 harbouring the binary vector pART27-10 to inoculate leaf strips. The vector contains neomycin phosphotransferase (nptII) and ${\beta}$-glucuronidase (GUS) (uidA) genes. A range of light intensities and different strengths of Murashige and Skoog (MS) basal salt media was used to overcome the problem of browning and/or necrosis of explants and calli. Callus browning was significantly reduced, resulting in regenerated adventitious shoots when the MS basal salt concentration in the culture medium was reduced to half-strength at low light intensity ($3.4\;{\mu}mol\;m^{-2}\;s^{-1}$) conditions. Inoculated leaf strips produced putative transformed shoots of Actinidia arguta on half-MS basal salt medium supplemented with 3.0 $mg\;l^{-1}$ zeatin, 0.5 $mg\;l^{-1}$ 6-benzyladenine, 0.05 $mg\;l^{-1}$ naphthalene acetic acid, 150 $mg\;l^{-1}$ kanamycin and 300 $mg\;l^{-1}$ $Timentin^{(R)}$. All regenerated plantlets were deemed putativ transgenic by histochemical GUS assay and polymerase chain-reaction analysis.

Genetic Transformation of Populus nigro x maximowiczii Using Agrobacterium tumefaciens Harboring Antisense OMT Gene

  • Ahn, In-Suk;Park, Young-Goo;Shin, Dong-Ill;Woo, Sahng-Young;Park, Hee-Sung;Park, Jang-Won;Sul, Ill-Whan
    • Journal of Plant Biotechnology
    • /
    • v.3 no.3
    • /
    • pp.135-140
    • /
    • 2001
  • An Agrobacterium tumefaciens LBA4404 (harboring antisense OMT gene)-mediated transformation method has been developed for poplar (P.nigra x maximowiczii) using prolonging co-cultivation time. Explants on LT (longterm) were induced transgenic calli one month earlier than those from ST (short-term) co-cultivation and remained healthier on LT than ST. With this approach, LT method reduced time to produce transgenic calli. Shoots were successfully regenerated from transgenic calli on SIM (Shoot Induction Medium) and rooted well on the basal medium spontaneously. The presence of antisense OMT gene was verified both by PCR and Southern analysis. Each transgenic poplar was phenotypically indishtinguishable when compared with controls for their growth pattern, leaf morphologies and xylem coloration.

  • PDF

Tobacco Transformation Using Expansin Genes (Expansin 유전자를 이용한 담배의 형질전환)

  • Choi Dongsu;Kim Ho-Bang;Kim Jeong-Hoe;Shin Ju-Sik;Seok Yeong-Seon;Chung Chan-Moon;Lee Yi
    • Journal of the Korean Society of Tobacco Science
    • /
    • v.27 no.2
    • /
    • pp.153-162
    • /
    • 2005
  • Tobacco (Nicotiana tabacum L.) cells were transformed with rice expansin genes, OsEXPA4, OsEXPB3, OsEXPB4, and OsEXPB6, to elucidate the function of the genes in tobacco cells. The transformation increased the mass of the callus by $36\%-65 \%$, and the cell length by $12\%-28\%$. The cell width was decreased by $3\%$ for OsEXPB3, not changed for OsEXPB4, increased by $25\%\;and\;20\%$ for OsEXPA4 and OsEXPB6, respectively. From database search, seven expansin genes were found and six of them belong to EXPA group and one of them belongs to EXPB group. EXLA and EXLB were not found. All tobacco expansin genes were evenly distributed in the phylogenetic tree of rice and Arabidopsis expansin genes.

Development of Basta Resistant Tobacco Using Artificial Phosphinothricin Acetyltransferase Gene (인공합성 Phosphinothricin Acetyltransferase 유전자에 의한 Basta 내성 연초식물체의 개발)

  • 양덕춘
    • Korean Journal of Plant Resources
    • /
    • v.11 no.2
    • /
    • pp.188-194
    • /
    • 1998
  • This experiment was conducted to introduce phosphinothricin acetyl -transferase(PAT) gene, resistant to basta and non-selective herbidide, into tobacco(Nicotiana tabacum cv.BY4). For shoot formation,tobacco leaf disks were placed on the MS medium supplemented with 2.0mg/L BA and 0.1mg/L NAA. In this medium condition, tobacco leaf disces were cocultivated with A. tumefaciens MP90 containing NPT IIand PAT resistant to kanamycin and Basta, respectively. Shoots were obtained in the medium containing antibiotics, and those were transferred to rooting medium supplemented with 0.1mg/L NAA and antibiotics. The plants obtaining roots were transplanted into soil. Phenotype of transgenic tobacco plant was mostly as normal plant. However, about 5% was abnormal plant, which did not set seeds. PCR analysis and southern blot were performed to determine transformation. As the results, it was confirmed that PAT gene was stably integrated into tobacco genome.When herbicide, basta, was sprayed to the plants confirmed by PCR, the transgenic plants showed normal growth, whereas normal plants died. Therefore, the result of this experiment show that tobacco transformation for the resistance to basta, non-selective herbicide, was successful because PAT gene was stably integrated into tobacco.

  • PDF