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- 저자 : Tripathi Sharanya (검색결과 3 건)
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Sharanya Tripathi,Jeong-Il Kim 전남대학교 농업과학기술연구소 2022 농업생명과학연구동향 Vol.60 No.-
Model plants are necessary and important for advances in various research areas of plant biology. Compared with Arabidopsis thaliana, a dicot model plant that is widely used thus far, monocot model plants are not popular in using plant biology research. In this regard, Brachypodium distachyon is suggested as a new and alternative monocot model plant for economically valuable temperate grasses and cereals such as wheat and barley, and also for bioenergy crops such as switchgrass and miscanthus. One of the major constraints in performing plant research would be the difficulty of genetic transformation and its low frequency. Here, we provide a detailed and improved method for Agrobacterium-mediated genetic transformation of B. distachyon, based on our experimental experience. With hygromycin resistance as a selectable marker, we could obtain approximately 40%−60% of the transformation efficiency using this method, in which the process takes approximately 26 weeks to obtain T1 seeds starting from co-cultivation of immature embryos with Agro- bacterium cells. Here, we also describe about important factors possibly contributing to the genetic transformation of B. distachyon.
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7종의 고추(Capsicum annuum L.) 재분화 조건 비교
김민수,한윤정,Sharanya Tripathi,곽진우,권진경,김정일 한국자원식물학회 2023 한국자원식물학회지 Vol.36 No.5
적 요고추(Capsicum annuum L.)는 전 세계적으로 재배되고 있는채소이자 향신료 작물이다. 고추 열매는 독특한 맛과 향을 지니면서 인체 건강에 중요한 항산화제와 다양한 화합물을 제공하기에 경제적 가치가 높다. 또한 소비자의 기호에 따른 새로운 고추 품종에 대한 수요도 높다. 하지만 고추는 조직 및 기관 분화와 식물 재분화가 어려운 식물이기에 유전자재조합 및 유전자교정과 같은 새로 개발된 기술을 사용하여 요구되는 품종 개발이 어려운 상황이다. 본 연구에서는 서울대학교 Core collection 에서 분양 받은 7계통의 고추를 이용하여 조직 배양 및 재분화조건을 조사하였다. 여기에는 서로 다른 사이토키닌과 옥신 농도로 구성된 여러 가지 배지 조성을 사용하여 캘러스와 부정아유도, 그리고 신초 형성을 관찰하였다. 그 결과, 호르몬 조성에따라 캘러스 유도 및 신초 형성에서 품종별 차이가 있는 것으로확인하였으며, zeatin riboside를 함유한 배지와 유묘의 잎자루부분을 사용한 경우 재분화율이 가장 높게 나타났다. 특히 사용된 7 계통의 고추 중에서 CMV980 계통이 다른 품종에 비해 월등히 높은 재분화 효율(약48%)을 나타냈고, Yuwolcho 계통이 그뒤를 이었다. 따라서 본 연구는 CMV980 및 Yuwolcho 계통을고추 형질전환에 활용될 수 있는 우수한 품종 후보로 제공하며, 이는 향후 유전자교정기술 등을 이용한 다양한 품종 개발에 기여할 수 있을 것이다. Abstract - Pepper (Capsicum annuum L.) is an important vegetable and spice crop that has been cultivated worldwide. Pepper fruits have unique taste and aroma, providing a variety of antioxidants and compounds important for human health, which makes a high economic value. In addition, there is a high demand for new pepper varieties, according to consumer’s preference. However, pepper is a recalcitrant plant for in vitro tissue and organ differentiation and plant regeneration, which makes it difficult to develop demanded varieties using newly developed technologies such as genetic engineering and gene editing. In this study, tissue culture and regeneration conditions were investigated using seven pepper varieties that were obtained from the core-collection of Seoul National University. We observed callus and bud induction and shoot formation using several media composition composed of different cytokinins and auxin concentrations. As a result, it was found that there were differences in callus induction and shoot formation of each variety depending on the hormone composition, and the highest regeneration was shown when the medium containing Zeatin Riboside and the petioles of seedlings were used. In particular, out of seven pepper varieties, CMV980 exhibited a higher regeneration efficiency (approximately 48%) than other varieties, followed by Yuwolcho. Therefore, this study provides CMV980 and Yuwolcho as good candidates that can be used for pepper transformation, which might contribute to the development of various varieties through gene editing technology in the future.
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Baeg Ga-Jin,김성현,최다민,Tripathi Sharanya,Han Yun-Jeong,Kim Jeong-Il 한국식물생명공학회 2021 Plant biotechnology reports Vol.15 No.6
Weeds are important biotic constraints to agricultural production, so herbicides are widely used with agronomic crops as the primary method of weed control. Accordingly, extensive efforts to develop herbicide-resistant (HR) crops have been made using various methods, including clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9)-mediated gene editing. Thus far, most of the HR crops developed using gene-editing methods have adopted site-specific mutations of endogenous genes, but rarely by insertion or deletion (indel) mutations. In this study, we attempted to confer herbicide resistance in plants via indel mutations using the CRISPR/Cas9-mediated editing of 5-oxoprolinase 1 (OXP1) gene as a target in Arabidopsis. OXP1 is an enzyme that is involved in 5-oxoproline metabolism and the glutathione recycling pathway. Here, we generated the indel mutants of Arabidopsis using OXP1 editing (oxp1/CRISPR), and verified a reduced sensitivity to sulfamethoxazole (Smex), a sulfonamide herbicide that suppresses the growth of cells by inhibiting the folic acid biosynthetic pathway. In addition, we successfully obtained OXP1-edited plants by screening the transgenic plants on Smex-containing media, suggesting the possible use of OXP1 as a marker for plant gene editing. Subsequent molecular analyses demonstrated the indel mutations on OXP1 in the Smex-resistant Arabidopsis plants. Moreover, the oxp1/CRISPR plants exhibited tolerance to heavy metals, such as cadmium, and another sulfonamide compound, amisulbrom. Therefore, the present study provides a way to confer resistance to sulfonamide compounds in plants by generating indel mutations.
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