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한지학 한국식물생명공학회 2006 JOURNAL OF PLANT BIOTECHNOLOGY Vol.33 No.3
According to ISAAA report, the global area of genetically modified (GM) crops increased more than 50 fold during the ten-year period from 1996 to 2005 with a sustained double-digit growth rate of 10%. This biotechnology adoption is one of the highest rates of technology adoption in agriculture history and this phenomenon indicates that the industrial value of the GM crops is highly perspective. In addition, the year 2010, 60% of cereal seeds in the global market would be GM or biotechnology related seeds so that the GM crop regards as the second green revolution that could provide a huge impact to food and agriculture. Nevertheless, there has not been any GM variety ever successfully commercialized in Korea and even none of the GM crops has ever been approved for safety testing by risk assessment. This seems that Korean agriculture industry might be indeed lost in the war of future seed market. However, lots of evidence show that Korean scientists have established advanced technologies and protocols to develop GM crops for last 20 years. Actually there have been many cases of successful transformation of crops that were previously known very difficult in transforming. Therefore, Korean agbiotechnology arena firmly holds an infrastructure for developing GM crops with a superior technology. Then what were the problems? Why has even a single GM crop not been commercialized in Korea? The tardiness shown by business in adopting the GM crop is caused by many factors: academical weakness, poor research funding, short knowledge of risk assessment, public concern, no successful experience, lack of professional leaders on GM variety development, lack of systems toward industrialization and inappropriate target transgenes from the beginning. In order to catch up in the race for the new green industry, each one of us in private sectors alongside academia and national research institutes needs to focus altogether on what can be done best in terms of choosing crops, investing fund and establishing a road map for commercialization of GM crops.
CGMMV-CP 형질전환 수박대목의 CGMMV 내성시험 및 계통확보
한지학,박상미,권정희,임미영,신윤섭,허남한,이장하,류기현 한국식물생명공학회 2007 JOURNAL OF PLANT BIOTECHNOLOGY Vol.34 No.1
Previously developed transgenic watermelon rootstocks (gongdae) inserted by CGMMV-CP were examined to test the virus tolerance levels. In the restricted plastic house, the T3 watermelon rootstock showed tolerance to CGMMV until 70 days after inoculation on the leaves while the non-transformed watermelon rootstock became susceptible at 20 days after inoculation. In the field, tolerance efficiency of transgenic rootstocks maintained up to 40% at 71 days after contamination with CGMMV in the soil while all of the non-transformed rootstocks became susceptible at 37 days with the same condition. In the same field, transgenic rootstocks showed more tolerance to CGMMV than the non-transformed rootstocks as those were inoculated on the leaves, but it showed only 10 days delay before being susceptible. Therefore, transgenic rootstocks have a characteristic of delay effect against CGMMV susceptibility, rather than resistance character. From T3 rootstocks homozygous for the CGMMV-CP, horticulturally favorable individuals were selected for further breeding and a transgenic line was finally obtained at the BC1T5. A material transfer experiment was conducted to find out if the DNA, RNA or expressed protein in the transgenic rootstocks could move to the grafted scion (non-transformed watermelon, Super-Kumcheon). PCR, northern, and western blot analysis were performed and no evidence of transferring of those materials from rootstock to scion was ever found.
한지학,권정희,박상미,임미영,신윤섭 한국식물생명공학회 2007 JOURNAL OF PLANT BIOTECHNOLOGY Vol.34 No.1
The genetic transformation of watermelon by Agrobacterium has been known very difficult and a few successful cases have been reported by obtaining the direct shoot formation. However, since this direct shoot formation is not guaranteed the stable transformation, the stable transformation with reproducibility is required by a different approach such as a callus induced manner. The best conditions for inducing the callus from cotyledon and root explants of watermelon were 2 mg/L zeatin + 0.1 mg/L IAA and 2 mg/L BA + 0.1 mg/L 2,4-D, respectively. The GFP expression in the callus was identified and monitored through fluorescent microscopy after transformation with pmGFP5-ER vector. Paromomycin rather than kanamycin was used for selecting the nptII gene expression because it was more effective to select the watermelon explants. Four different callus types were observed and the solid green callus showed stronger GFP expression. The highest frequency of GFP expression in the callus developed from cotyledon was 9.0% (WM8 inbred line), while the highest frequency from root was 8.3% (WM6 inbred line). The WMV-CP was transformed using the method of GFP transformation and the genetic transformation of WMV-CP was confirmed by PCR and Southern blot analysis. Here we present a system for callus induction of watermelon explant and the callus induced method would facilitate the establishment of stable watermelon transformation.