• Journal of Plant Biotechnology
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• v.33 no.1
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• pp.19-25
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• 2006

With the use of Agrobacterium and gene-gun, cold regulated gene (BN115) has been injected in Chrysanthemum leaf disc and transgenic plants have been produced successfully on the selection media containing phytohormone. To determine the presence of the transferred cold regulated gene (BN115) in the transgenic Chrysanthemum, PCR-amplification indicated the presence of that gene. Real-Time PCR for confirmation of the putative transgenic plants was established. The copy number of cold regulated gene (BN115) is extrapolated on the basis of a standard curve. Serial dilutions of known number of gene copies were in triplicates. In this diagram, PCR cycles are plotted against the fluorescence intensity. The cycle at which the fluorescence reaches a threshold cycle is inversely proportional to the starting amount of target DNA.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.40 no.1
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• pp.69-76
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• 1995

Cold acclimation involves changes in gene expression. BN28 and BN115 are two genes which are regulated by cold temperature and assumed having roles in cold acclimation. The objectives of this experiment was to explore the expression of BN28 and BN115 under field conditions. Six winter cultivars were planted at three different dates during the fall. The expression of the genes was determined by northern blot analysis of total RNA taken from leaves 15 to 30 day-intervals after planting. The expression of the two genes was detected within 15 days after planting well before onset of freezing tolerance in plants. This suggestes either their expression was a prerequisite of the freezing tolerance or their expression was regulated by other environmental factors as well as temperature. Two genes showed a different expression pattern suggesting they had a different regulatory system. Although timecourse increase in expression of the cold-regulated genes was matched with increase in freezing tolerance, the difference of expression in cultivar level at specific times of measurement was not correlated with freezing tolerance at the moment.

• Korean Journal of Plant Tissue Culture
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• v.27 no.1
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• pp.7-12
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• 2000

Explants of lettuce (Lactuca sativa L.) were co-cultivated with Agrobacterium tumifacience GV 3101 strain containing nptII gene and cold regulated gene (BN115) from Brassica napus for transformation. Multiple shoots were obtained from the explants in the selection medium (MS basal medium supplemented with 100 mg/L kanamycin, 500 mg/L carbenicillin, 0.1 mg/L NAA, 0.5 mg/L kinetin) after 3 to 4 weeks of co-culture. The putative transgenic shoots were transferred to rooting medium (1/2 MS basal medium supplemented with 100 mg/L kanamycin and 250 mg/L carbenicillin). The selected shoots were tested with PCR analysis using nptll, BN115 primers whether cold-regulated gene was introduced to genome of the plants. The vir G primers were particularly used to check contamination of Agrobacterium during PCR analysis. The nptII and BN115 primers produced the specific PCR bands in the putative transgenic lines but the vir G primers did not. These results confirmed that the PCR products were not the result of contamination with Agrobacterium. Additionally the Southern analysis of the PCR products and RT-PCR analysis proved that the cold-regulated gene was successfully integrated and transcribed in the putative transgenic lettuce plants.

• Journal of Plant Biotechnology
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• v.35 no.1
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• pp.55-61
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• 2008

Previous studies on genetic transformation of chrysanthemum using cold regulated gene (BN115) have been conducted and the PCR and Real-Time PCR based method to determine the presence of the transferred cold regulated gene in the chrysanthemum was established. To check whether over-expression of BN115 gene in transgenic chrysanthemum will enhance their tolerance to cold stress, the transgenic chrysanthemum were grown under low temperature condition and several cold signalling including growth characteristics, stoma size and shape, SPAD value and ion leakage test were investigated. The transgenic chrysanthemum in the low temperature growth chamber grow much faster in term of the height, number and size of the leaves than those of wild-type plants and damage of transgenic plant caused by the low temperature was much less than that of wild-type plants. The stoma type and size of transgenic plant leaves grown at $5^{\circ}C$ were much similar to of wild-type plant cultured on $25^{\circ}C$ It has been found that SPAD value of transgenic plants was much higher than those of wild-type, but the EC density being lower under low temperature condition.