• The Plant Pathology Journal
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• v.32 no.2
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• pp.112-122
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• 2016

Virus-induced gene silencing (VIGS) is an effective tool for the study of soybean gene function. Successful VIGS depends on the interaction between virus spread and plant growth, which can be influenced by environmental conditions. Recently, we developed a new VIGS system derived from the Soybean yellow common mosaic virus (SYCMV). Here, we investigated several environmental and developmental factors to improve the efficiency of a SYCMV-based VIGS system to optimize the functional analysis of the soybean. Following SYCMV: Glycine max-phytoene desaturase (GmPDS) infiltration, we investigated the effect of photoperiod, inoculation time, concentration of Agrobacterium inoculm, and growth temperature on VIGS efficiency. In addition, the relative expression of GmPDS between non-silenced and silenced plants was measured by qRT-PCR. We found that gene silencing efficiency was highest at a photoperiod of 16/8 h (light/dark) at a growth temperature of approximately $27^{\circ}C$ following syringe infiltration to unrolled unifoliolate leaves in cotyledon stage with a final SYCMV:GmPDS optimal density $(OD)_{600}$ of 2.0. Using this optimized protocol, we achieved high efficiency of GmPDS-silencing in various soybean germplasms including cultivated and wild soybeans. We also confirmed that VIGS occurred in the entire plant, including the root, stem, leaves, and flowers, and could transmit GmPDS to other soybean germplasms via mechanical inoculation. This optimized protocol using a SYCMV-based VIGS system in the soybean should provide a fast and effective method to elucidate gene functions and for use in large-scale screening experiments.

• The Bulletin of The Korean Astronomical Society
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• v.16
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• pp.20.1-20.1
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• 1991

• The Plant Pathology Journal
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• v.21 no.2
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• pp.158-163
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• 2005

Virus-induced gene silencing (VIGS) is a recently developed gene transcript suppression technique for characterizing the function of plant genes. However, efficient VIGS has only been studied in a few plant species. In order to extend the application of VIGS, we examined whether a VIGS vector based on TRV would produce recognizable phenotypes in soybean. Here, we report that VIGS using the Tobacco rattle virus (TRV) viral vector can be used in several soybean cultivars employing various agro-inoculation methods including leaf infiltration, spray inoculation, and agrodrench. cDNA fragments of the soybean phytoene desaturase(PDS) was inserted into TRV RNA-2 vector. By agrodrench, we successfully silenced the expression of PDS encoding gene in soybean. The silenced phenotype of PDS was invariably obvious 3 weeks after inoculation with the TRV-based vector. Real-time RT-PCR analyses showed that the endogenous level of GmPDS transcripts was dramatically reduced in the silenced leaf tissues. These observations confirm that the silenced phenotype is closely correlated with the pattern of tissue expression. The TRV-based VIGS using agrodrench can be applied to functional genomics in a soybean plants to study genes involved in a wide range of biological processes. To our knowledge, this is the first high frequency VIGS method in soybean plants.

• Journal of Technologic Dentistry
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• v.36 no.3
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• pp.141-147
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• 2014

Purpose: Dental casting #Gr I (Co-25Cr-5Mo-3Sn-1Mn-1Si), #Gr II (Co-25Cr-5Mo-5Cu-1Mn -1Si) and #Gr III (Co-25Cr-5Mo-3Sn-5Cu-1Mn-1Si) master alloys of granule type were manufactured the same as manufacturing processes for dental casting Ni-Cr and Co-Cr-Mo based alloys of ingot type. These alloys were analyzed melting processes with heating time of high frequency induction centrifugal casting machine using infrared thermal image analyzer. Methods: These alloys were manufactured such as; alloy design, the first master alloy manufatured using vacuum arc casting machine, melting metal setting in crucible, melting in VIM, pouring in the mold of bar type, cutting the gate and runner bar and polishing. These alloys were put about 30g/charge in the ceramic crucible of high frequency induction centrifugal casting machine and heat, Infrared thermal image analyzer indicated alloys in the crucible were set and operated. Results: The melting temperatures of these alloys measuring infrared thermal image analyzer were decreased in comparison with remanium$^{(R)}$ GM 800+, vera PDI$^{TM}$, Biosil$^{(R)}$ f, WISIL$^{(R)}$ M type V, Ticonium 2000 alloys of ingot type and vera PDS$^{TM}$(Aabadent, USA), Regalloy alloys of shot type. Conclusion: Co-Cr-Mo based alloy in addition to Sn(#Gr I alloy) were decreased the melting temperature with heating time of high frequency induction centrifugal casting machine using infrared thermal image analyzer.