• 제목/요약/키워드: Bar gene

검색결과 102건 처리시간 0.039초

Production of transgenic Alstroemeria plants containing virus resistance genes via particle bombardment

  • Kim, Jong Bo
    • Journal of Plant Biotechnology
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    • 제47권2호
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    • pp.164-171
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    • 2020
  • Transgenic Alstroemeria plants resistant to Alstroemeria mosaic virus (AlMV) were generated through RNA-mediated resistance. To this end, the friable embryogenic callus (FEC) of Alstroemeria was induced from the leaf axil tissue and transformed with a DNA fragment containing the coat protein gene and 3'-nontranslated region of AlMV through an improved particle bombardment system. The bar gene was used as a selection marker. More than 300 independent transgenic FEC lines were obtained. Among these, 155 lines resistant to phosphinothricin (PPT) were selected under low stringent conditions. After increasing the stringency of PPT selection, 44 transgenic lines remained, and 710 somatic embryos from these lines germinated and developed into shoots. These transgenic shoots were then transferred to the greenhouse and challenged with AlMV. In total, 25 of the 44 lines showed some degree of resistance. PCR analysis confirmed the presence of the viral sequence. Virus resistance was observed at various levels. Establishment of an efficient transformation system for Alstroemeria will allow inserting transgenes into this plant to confer resistance to viral and fungal pathogens. Accordingly, this is the first report on the production of a transgenic virus-resistant Alstroemeria and lays the foundation for alternative management of viral diseases in this plant.

Organogenesis and Production of Some Transgenic Legume Plants by Agrobacterium tumefaciens-mediated Herbicide Resistance Gene Transformation

  • Kantayos, Vipada;Lee, Hyo-Yeon;Bae, Chang-Hyu
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2018년도 추계학술대회
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    • pp.52-52
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    • 2018
  • Development of herbicide resistant transgenic legume plants through Agrobacterium-mediated transformation has been worked in many previous studied. Plant regeneration after infection is the important step to obtain successful transgenic plants. Many attempts try to find the optimum media condition for plant regeneration after infection. However, the transformation efficiency of legume plants is still low. In this study, regeneration of some Korean legume species including two soybean cultivars (Dawon and Pungsan) and pea have been done with organogenesis which is used various kind of explants such as cotyledonary-nodes in soybean and bud-containing tissue in pea. We developed the optimum media condition for plant regeneration regulators under Agrobacterium-mediated transformation using different kind and various concentration of plant growth. As the results, B5 medium containing 2 mg/L of 6-benzylaminopurine was selected in this study for the optimum plant regeneration media. The segments were inoculated with Agrobacterium suspension harbored an IG2 vector containing bar gene which confers resistance to phosphinotricin (PPT) in 3, 5 and 7 days. The transformation efficiency was achieved in Dawon 3.03 % and pea 1.46 % with co-cultivation period of 7 days which is showed a high number of GUS positive expression period.

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Study about gene flow and stability assessment in GM rice (Oryza sativa L.)

  • Kim, Jun-Hoi;Kim, Kyung-Min
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
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    • pp.136-136
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    • 2017
  • In agriculture, the rice is the one of important things. Many farmers and scientists have long tried to increase the yield of rice. So many technologies have been developed these days. One type of technology has given rise to a host of concerns and questions, namely Genetically Modified Organisms (GMOs). The increasing cultivation of GM crops has raised a wide range of concerns with respect to food safety, environmental effects and socio-economic issues and now commercially planted on about 100 million hectares in some 22 developed and developing countries. The scientific evidence concerning the environmental and health impacts of GMOs is still emerging, but so far there is no conclusive information on the definitive negative impacts of GMOs on health or the environment. Nevertheless, public perceptions about GMOs in food and agriculture are divided with a tendency toward avoiding GM food and products in many developed and developing countries. Also Korea is one of that country and is not allow the GMOs now. So I studied whether these GMOs are actually dangerous for environment and whether there are differences in cultivar characteristics such as germination test with TTC tetrazolium, germination test in frozen soil and gene-flow test with glufosinate and strip-bar test. With these experiments, we evaluated the agricultural safety of GM rice and to identify and assess environmental risks.

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BLOOD PROTEIN POLYMORPHISMS OF NATIVE AND JUNGLE FOWLS IN INDONESIA

  • Hashiguchi, T.;Nishida, T.;Hayashi, Y.;Maeda, Y.;Mansjoer, S.S.
    • Asian-Australasian Journal of Animal Sciences
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    • 제6권1호
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    • pp.27-35
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    • 1993
  • In an attempt to reveal the interrelationship between fowls of jungle and native origin, their gene constitutions were compared using gene frequencies at the 16 loci controlling blood protein variations. Of the 16 loci analysed by electrophoresis, polymorphism was detected at following seven loci: Es-1, Amy-1, Akp-akp, Akp-2, Alb, Tf and 6-PGD. The other nine loci: Amy-3, Es-D, PGM, PHI, MDH, To, LDH, Hb-1 and Hb-2, were noted to be monomorphic. Genetic distance between pairs of native fowl and jungle fowls was estimated by a numerical taxonomic method. The Indonesian native fowl was genetically close to the Indonesian red jungle fowl, and the grey jungle fowl was genetically similar to the Ceylonese jungle fowl. It was also suggested that the green jungle fowl was genetically remote from the other jungle fowls and from the Indonesian native flow. The proportion of polymorphic loci (Ppoly), the expected average heterozygosity per individual $\bar{H}$, and the effective number of alleles per locus (Ne) were calculated to evaluate the genetic variabilities in the native and jungle fowls. The Indonesian native fowl exhibited slightly higher the proportion of polymorphic loci than the jungle fowls.

Establishment of a Simple and Rapid Gene Delivery System for Cucurbits by Using Engineered Zucchini Yellow Mosaic Virus

  • Kang, Minji;Seo, Jang Kyun;Choi, Hoseong;Choi, Hong Soo;Kim, Kook Hyung
    • The Plant Pathology Journal
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    • 제32권1호
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    • pp.70-76
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    • 2016
  • The infectious full-length cDNA clone of zucchini yellow mosaic virus (ZYMV) isolate PA (pZYMV-PA), which was isolated from pumpkin, was constructed by utilizing viral transcription and processing signals to produce infectious in vivo transcripts. Simple rub-inoculation of plasmid DNAs of pZYMV-PA was successful to cause infection of zucchini plants (Cucurbita pepo L.). We further engineered this infectious cDNA clone of ZYMV as a viral vector for systemic expression of heterologous proteins in cucurbits. We successfully expressed two reporter genes including gfp and bar in zucchini plants by simple rub-inoculation of plasmid DNAs of the ZYMV-based expression constructs. Our method of the ZYMV-based viral vector in association with the simple rub-inoculation provides an easy and rapid approach for introduction and evaluation of heterologous genes in cucurbits.

Overexpression of a Chromatin Architecture-Controlling ATPG7 has Positive Effect on Yield Components in Transgenic Soybean

  • Kim, Hye Jeong;Cho, Hyun Suk;Pak, Jun Hun;Kim, Kook Jin;Lee, Dong Hee;Chung, Young-Soo
    • Plant Breeding and Biotechnology
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    • 제5권3호
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    • pp.237-242
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    • 2017
  • AT-hook proteins of plant have shown to be involved in growth and development through the modification of chromatin architecture to co-regulate transcription of genes. Recently, many genes encoding AT-hook protein have been identified and their involvement in senescence delay is investigated. In this study, soybean transgenic plants overexpressing chromatin architecture-controlling ATPG7 gene was produced by Agrobacterium-mediated transformation and investigated for the positive effect on the important agronomic traits mainly focusing on yield-related components. A total of 27 transgenic soybean plants were produced from about 400 explants. $T_1$ seeds were harvested from all transgenic plants. In the analysis of genomic DNAs from soybean transformants, ATPG7 and Bar fragments were amplified as expected, 975 bp and 408 bp in size, respectively. And also exact gene expression was confirmed by reverse transcriptase-PCR (RT-PCR) from transgenic line #6, #7 and #8. In a field evaluation of yield components of ATPG7 transgenic plants ($T_3$), higher plant height, more of pod number and greater average total seed weight were observed with statistical significance. The results of this study indicate that the introduction of ATPG7 gene in soybean may have the positive effect on yield components.

GMO 격리포장에서의 유전자변형 들잔디로부터 토착미생물로의 수평유전자전달 평가 (Evaluation of horizontal gene transfer from genetically modified zoysiagrass to the indigenous microorganisms in isolated GMO field)

  • 배태웅;이효연;류기현;이태형;임평옥;윤필용;박신영;류기중;송필순;이용억
    • Journal of Plant Biotechnology
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    • 제34권1호
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    • pp.75-80
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    • 2007
  • The release of genetically modified organisms ($GMO_{s}$) into the environment has the potential risks regarding the possibility of gene transfer from $GMO_{s}$ to natural organisms and this needs to be evaluated. This study was conducted to monitor the possible horizontal gene transfer from herbicide-resistant zoysiagrass (Zoysia japonica Steud.) to indigenous microorganisms. We have first examined the effect of field-released GM zoysiagrass on the microbial flora in the gut of locust (Locusts mlgratoria). The microbial flora was analyzed through determining the 165 rDHA sequences of microorganisms. The comparison of the microbial flora in the gut of locusts that were captured at the field of GM zoysiagrass and of wild-type revealed that there is no noticeable difference between these two groups. This result indicates that the GM zoysiagrass does not have negative impact on microbial flora in the gut of locust. We then investigated whether the horizontal gene transfer occurred from GM zoysiagrass to microbes in soil, rhizosphere and faecal pellets from locusts by utilizing molecular tools such as Southern hybridization and polymerase chain reaction (PCR). When the total DNAs isolated from microbes in GM zoysiagrass and in wild-type zoysiagrass fields were hybridized with probes for bar or hpt gene, no hybridization signal was detected from both field isolates, while the probes were hybridized with DNA from the positive control. Absence of these genes in the FNAs of soil microorganisms as well as microbes in the gut of locust was further confirmed by PCR. Taken together, our data showed that horizontal gene transfer did not occur in this system. These results further indicate that frequencies of transfer of engineered plant DNA to bacteria are likely to be negligible.

Blood Protein Polymorphisms of Native Fowls in Laos

  • Okamoto, S.;Tsunekawa, N.;Kawamoto, Y.;Worawut, R.;Kawabe, K.;Maeda, Y.;Nishida, T.
    • Asian-Australasian Journal of Animal Sciences
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    • 제12권7호
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    • pp.1011-1014
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    • 1999
  • Blood protein polymorphism of fowls in Laos was analyzed by electrophoresis. Blood samples were collected in the area of Viangchan, Louangphrabang and Pakxe. Out of 17 loci, polymorphism was detected at the following seven loci; ES-1, Amy-1, Akp-akp, Akp-2, Alb, Tf and Pas. The other ten loci; Amy-3, LDH, 6-PGD, PGM, PHI, To, MDH, Es-D, Hb-l, Hb-2 were noted to be monomorphic. The proportion of polymorphic loci $(P_{poly})$, the expected average heterozygosity per individual ($\bar{H}$), and the subdivision index $(G_{ST})$ of the native fowl in Laos was $0.412{\pm}0.123$, 0.106 and 0.026, respectively. Genetic distance between native fowls in Laos, Bangladesh, and Nepal was clustered in one group.