Suppression of Phytophthora infestans via exogenous dsRNA-induced gene silencing

Yujin Kweon,Dowhan Lee,Chanseok Shin 한국육종학회 2023 한국육종학회 공동학술발표집 Vol.2023 No.-

Sequence optimization and multiple gene-targeting improve the inhibitory efficacy of exogenous double-stranded RNA against pepper mottle virus in Nicotiana benthamiana

Kweon Yujin,Fang Miao,Shin Sang-Yoon,Lee Dowhan,Kim Kook-Hyung,Shin Chanseok 한국응용생명화학회 2022 Applied Biological Chemistry (Appl Biol Chem) Vol.65 No.6

Double-stranded RNA (dsRNA)-induced RNA interference is a promising agricultural technology for crop protection against various pathogens. Recent advances in this field have enhanced the overall efficiency with which this approach inhibits pathogenic viruses. Our previous study verified that treatment of Nicotiana benthamiana plants with dsRNAs targeting helper component-proteinase (HC-Pro) and nuclear inclusion b (NIb) genes protected the plant from pepper mottle virus (PepMoV) infection. The aim of this study was to improve the inhibitory efficacy of dsRNAs by optimizing the target sequences and their length and by targeting multiple genes via co-treatment of dsRNAs. Each of the two targeting dsRNAs were divided into three shorter compartments and we found that HC-Pro:mid- 1st and NIb:mid-3rd showed significantly superior antiviral potency than the other fragments, including the parent dsRNA. In addition, we confirmed that the co-treatment of two dsRNAs targeting HC-Pro and NIb produced a greater inhibition of PepMoV replication than that obtained from individual dsRNA treatment. Complementing our previous study, this study will provide future directions for designing dsRNAs and enhancing their efficiency in dsRNA-mediated RNA interference technologies. Double-stranded RNA (dsRNA)-induced RNA interference is a promising agricultural technology for crop protection against various pathogens. Recent advances in this field have enhanced the overall efficiency with which this approach inhibits pathogenic viruses. Our previous study verified that treatment of Nicotiana benthamiana plants with dsRNAs targeting helper component-proteinase ( HC-Pro ) and nuclear inclusion b ( NIb ) genes protected the plant from pepper mottle virus (PepMoV) infection. The aim of this study was to improve the inhibitory efficacy of dsRNAs by optimizing the target sequences and their length and by targeting multiple genes via co-treatment of dsRNAs. Each of the two targeting dsRNAs were divided into three shorter compartments and we found that HC-Pro :mid-1st and NIb :mid-3rd showed significantly superior antiviral potency than the other fragments, including the parent dsRNA. In addition, we confirmed that the co-treatment of two dsRNAs targeting HC-Pro and NIb produced a greater inhibition of PepMoV replication than that obtained from individual dsRNA treatment. Complementing our previous study, this study will provide future directions for designing dsRNAs and enhancing their efficiency in dsRNA-mediated RNA interference technologies.

Breeding of multi-viral resistance tomato by CRISPR/Cas9 genome-editing technology

Yujin Kweon,Sang-Yoon Shin,Minsu Park,Minsun Oh,Chanseok Shin 한국육종학회 2023 한국육종학회 공동학술발표집 Vol.2023 No.-

알러지 저감밀 오프리 밀가루의 품질 특성과 제빵 및 제면 적성

문유진 ( Yujin Moon ),조혜진 ( Hyejin Cho ),백나연 ( Nayeon Baek ),샤톈이 ( Tianyi Xia ),권미라 ( Meera Kweon ) 한국식품조리과학회 2021 한국식품조리과학회지 Vol.37 No.6

Purpose: This study aimed to explore the quality characteristics and bread-making and noodle-making properties of low allergy wheat (‘O-free’) flour. Methods: ‘O-free’ flour and three commercial wheat flours (high-H, medium-M, low-L gluten strength) were used. Solvent retention capacity (SRC), sodium dodecyl sulfate (SDS)-sedimentation volume, pasting and gelatinization properties using rapid visco analyzer and differential scanning calorimetry, and dough mixing property using a mixograph of the flour samples were determined. In addition, their bread- and noodle-making performance was evaluated. Results: The SRC values in all four solvents and the SDS-sedimentation volume of ‘O-free’ flour were lower than those of H and M flour and higher than those of L flour. The pasting and gelatinization properties of ‘O-free’ flour starch were similar to those of the other flour samples. The dough mixing property and bread-making performance of the ‘O-free’ flour were inferior to the H and M flour but superior to the L flour. On the other hand, the noodle-making performance of the ‘O-free’ flour was inferior to all three flours. Conclusions: Overall, it was confirmed that ‘O-free’ flour, which lacks omega-gliadin, has inferior gluten protein quality and processing performance compared to H or M flours that are mainly used for making bread and noodles. Performance improvement was necessary by controlling other basic ingredients and processing conditions.

시중 국산밀 및 외국밀 통밀가루의 입자크기와 품질 특성 비교

문유진 ( Yujin Moon ),샤톈이 ( Tianyi Xia ),조혜진 ( Hyejin Cho ),권미라 ( Meera Kweon ) 한국식품조리과학회(구 한국조리과학회) 2021 한국식품조리과학회지 Vol.37 No.1

Purpose: The particle size and quality characteristics of commercial whole wheat flours made of Korean domestic wheat and foreign wheat were compared. Methods: Two groups of whole wheat flours [1) eight different whole wheat flours manufactured with Korean domestic wheat by three Korean millers, and 2) eight different flours with foreign wheat by one Korean miller and five foreign millers] were used in the study. Whole wheat flour was analyzed by particle size, color, ash and damaged starch contents, solvent retention capacity (SRC), SDS sedimentation, and RVA pasting property. Results: The particle size, color, and pasting viscosities of whole wheat flours were significantly different between the two groups. In contrast, the ash and damaged starch contents, water and sodium carbonate SRC, and SDS sedimentation values were not significantly different between the groups. However, all measured parameters were significantly different between the products within each group. In particular, the particle size correlated negatively with the color L* and positively with a*. Conclusion: The wheat origin, such as Korean or foreign wheat, and millers, including milling methods, significantly affect the quality of the whole wheat flours. The particle size was likely a significant factor related to the quality variation of commercial whole wheat flours, which needs specifying and controlling for maintaining the quality of whole wheat flour.

요인설계법을 활용하여 자색밀기울 혼합 국산밀가루로 제조한 빵의 주요 배합비 및 공정 요인 모색

문유진 ( Yujin Moon ),서예리 ( Yeri Seo ),김경훈 ( Kyeong Hoon Kim ),권미라 ( Meera Kweon ) 한국식품조리과학회(구 한국조리과학회) 2021 한국식품조리과학회지 Vol.37 No.2

Purpose: This study aimed to identify significant factors for the processing conditions of bread formulated with flour-bran blends with purple-colored wheat (Ariheuk) bran to improve bread quality. Methods: The solvent retention capacity (SRC) and SDS sedimentation volume of the flour-bran blends at different blending ratios were analyzed. Bread baking with the blends using a factorial design was conducted, and the bread quality was measured. Water amount, mixing time, and fermentation time were chosen as factors, and the two levels of each factor were as follows: 55 and 75 g water; 1.5 and 3.5 min mixing time; 50 and 70 min fermentation time. Results: The water and sodium carbonate SRC values of the flour-bran blends increased significantly with increasing bran blending ratio (10, 20, and 30%), and the SDS sedimentation volume of the blends decreased significantly. The effects on moisture loss during baking were the most significant with water amount, followed by fermentation time. The effects on bread volume and height were most significant with mixing time. The favored formula and processing conditions for bread prepared with the flour-bran blend at 30% bran, which had the highest volume and height, were 75 g water, 3.5 min mixing time, and 60 min fermentation time. Conclusion: Overall, purple-colored wheat bran can be blended for producing bread with improved quality by controlling the basic formula and processing conditions.

시판 국산밀가루의 품질특성과 건면 제조적성

문유진(Yujin Moon),전수정(Soojeong Jeon),홍예은(Ye Eun Hong),쟈오위샤(Yuxia Zhao),김경훈(Kyeong Hoon Kim),권미라(Meera Kweon) 한국식품조리과학회 2018 한국식품조리과학회지 Vol.34 No.4

Purpose: This study assessed the quality of commercial Korean domestic wheat flours and their dry noodle-making performance. Methods: Six commercial Korean domestic wheat flours produced from different millers and a commercial all-purpose flour as a control were analyzed for moisture, ash and protein contents, solvent retention capacity (SRC), mixograph, and noodle making performance of dry noodles. Results: The moisture content of the commercial Korean domestic wheat flours varied according to the millers. The ash content of them was significantly higher than that of commercial all-purpose flour. The SRC results showed all Korean domestic flours had much lower lactic acid SRC values and gluten performance index (GPI) than the control. Mixograph data showed that one of the commercial Korean domestic flours (flour D) showed a very narrow bandwidth due to the higher damaged starch and arabinoxylan contribution based on high sodium carbonate and sucrose SRC values. Dry noodles made with commercial Korean domestic wheat flours produced significantly higher turbidity of cooked water and weight gain of cooked noodle than that with the control flour. In addition, the texture of cooked noodles made with commercial Korean domestic wheat flours was less viscoelastic, as illustrated by the less springiness, cohesiveness, gumminess and chewiness, compared to the control. Conclusion: Overall, the dry noodle-making performance of commercial Korean domestic wheat flours was inferior to that of commercial all-purpose flour due to the higher ash content and lower GPI. Lowering milling extraction and application of enzymes for reducing water absorption capacity of flour might improve the dry noodle-making performance.

Non-GMO 고아밀로스 밀가루의 전분관련 특성 연구

문유진 ( Yujin Moon ),권미라 ( Meera Kweon ) 한국식품조리과학회 2021 한국식품조리과학회지 Vol.37 No.6

Purpose: This study aimed to assess the effects of the amylose content of wheat on flour quality characteristics with a focus on starch-related properties. Methods: Three varieties of wheat flour (W, N, and H flour) milled from waxy, normal, and non-GMO high amylose wheat, respectively, were used for analyzing the amylose and total starch contents, pasting and gelatinization properties using rapid visco analyzer and differential scanning calorimetry, and the crystallinity using an X-ray diffractometer. In addition, the in vitro starch digestibility of the flour samples was measured. Results: Compared to the N and W flour, the pasting viscosity of the H flour little increased by suppressed swelling of starch granules. H flour gelatinized at a higher temperature with a smaller enthalpy than the N and W flour. The crystallinity pattern in H flour was observed as B-type, compared to the N and W flours as A-types. The rapidly digestible starch and slowly digestible starch contents of the H flour were significantly lower than those of the N and W flour, but the resistant starch content was significantly higher. Conclusions: Overall results confirmed that the H flour could be successfully used as an ingredient for resistant starch. Besides, H flour can be used without dilution of gluten in bread or noodle making, which would be advantageous compared to commercially available resistant starch ingredients.

Development of multi-target dsRNAs targeting PcNLP gene family to suppress Phytophthora capsici infection in Nicotiana benthamiana

Park Minsu,Kweon Yujin,Eom Jihyun,Oh Minsun,Shin Chanseok 한국응용생명화학회 2023 Applied Biological Chemistry (Appl Biol Chem) Vol.66 No.-

Phytophthora capsici, which causes diseases in solanaceous crops, secretes necrosis and ethylene-inducing peptide 1-like proteins (NLPs) that induce plant defense responses and leaf necrosis. In this study, we used RNA interference (RNAi) technique, a proven strategy for crop protection and gene regulation in plants, to suppress P. capsici infection through the inhibition of PcNLPs. In the RNAi mechanism, Dicer processes double-stranded RNA (dsRNA) into smaller entities known as small interfering RNAs (siRNAs). These siRNAs subsequently integrate into the RNA-induced silencing complex to form sequence-specific base pairing with complementary regions of the target mRNA. This interaction effectively initiates the degradation process of the target mRNA. We designed and synthesized dsRNAs targeting the “AIMY” and “GHRHDWE” conserved motifs of PcNLP gene family, which are predicted to be key elements for the expression of NLPs and pathogen infection. After infiltration of dsRNAs targeting the motifs and inoculation with P. capsici, we confirmed a significant suppression of P. capsici infection and downregulation of the PcNLP gene family. These findings imply that the dsRNA-mediated RNAi technique holds potential for mitigating a wide range of pathogens, while simultaneously suppressing the expression of a particular gene family using dsRNA targeting functional conserved motifs in the gene family.

Suppression of Phytophthora capsici using double-stranded RNAs targeting NLP effector genes in Nicotiana benthamiana

Park Minsu,Kweon Yujin,Lee Dowhan,Shin Chanseok 한국응용생명화학회 2023 Applied Biological Chemistry (Appl Biol Chem) Vol.66 No.-

RNA interference (RNAi) is a gene regulatory mechanism that involves the interaction of small interfering RNAs (siRNAs) and RNA-induced silencing complex (RISC). Dicer cleaves exogenous double-stranded RNA (dsRNA) into siRNAs, which get incorporated into RISC and bind to complementary sequences on the target mRNA to induce its degradation. In this study, we adopted RNAi technology using dsRNAs to suppress Phytophthora capsici, which causes diseases in solanaceous crops, including pepper. We designed and synthesized dsRNAs targeting the P. capsici effector genes PcNLP2 and PcNLP6, respectively. These genes encode necrosis and ethylene-inducing peptide 1-like proteins in P. capsici, which are known to promote oomycete infection. Nicotiana benthamiana leaves were first infiltrated with dsRNAs and inoculated with P. capsici 2 days later. We confirmed significant suppression of P. capsici and PcNLP2, PcNLP6 expression in dsRNA-treated leaves. In addition, we found that downregulation of PcNLP2 and PcNLP6 distinctly affected the expression of some defense-related genes. These results suggest that dsRNA mediated RNAi technology can be used to suppress various pathogens, and may contribute toward crop protection.