Movement of Metal Nanoparticle and GNP mediated Photothermal Impact in Arabidopsis

구연종 ( Yeonjong Koo ),자넷부라암 ( Janetbraam ) 한국환경농학회 2018 한국환경농학회 학술대회집 Vol.2018 No.-

The basic information to develop nanoparticle as a functional material is discovering nanoparticle movement in plant tissues. Two different methods were carried out to tracking nanoparticle in Arabidopsis. First, Arabidopsis was exposed to CdSe/CdZnS QDs with three different coatings, anionic, cationic, and relatively neutral, and nanoparticle movement was dramatically changed from the particle absorption into the plant root cell to the particle localization in plant leaves. T. ni caterpillars that fed on Arabidopsis exposed to QDs had reduced performance, and QD fluorescence was detected in both T. ni bodies and frass, demonstrating trophic transfer of intact QDs from plants to insects. Second, gold nanoparticle was applied to test the biological responses to photothermal effects of nanoparticles. The uptake of gold nanoparticle through Arabidopsis roots and translocation to leaves are reported through the photoacoustic signal detection. Furthermore, Arabidopsis leaves harboring GNPs and exposed to continuous laser or noncoherent light show elevated temperatures across the leaf surface and induced expression of heat-shock regulated genes. Overall, these results demonstrate that metal based nanoparticles are strong candidate as a substance carrier for manipulating plant physiology.

후숙 조절 유전자 Pectate lyase와 Phytoene Synthase 편집용 CRISPR-Cas9 sgRNA의 유전자 편집 효율 측정

박효선 ( Hyosun Park ),양소희 ( So Hee Yang ),김의연 ( Euyeon Kim ),구연종 ( Yeonjong Koo ) 한국환경농학회 2021 한국환경농학회지 Vol.40 No.3

BACKGROUND: Tomato genome editing using CRISPR-Cas9 is being actively conducted in recent days, and lots of plant researches have been aiming to develop high valued crops by editing target genes without inserting foreign genes. Many researchers have been involved in the manipulation of the crop ripening process because fruit ripening is an important fruit phenotype for increasing fruit shelf life, taste, and texture of crops. This paper intends to evaluate target sgRNA to edit the two ripening-related genes encoding pectate lyase (PL) and phytoene synthase (Psy) with the CRISPR-Cas9 system. METHODS AND RESULTS: The CRISPR-Cas9 expression vector was cloned to target the PL (Solyc03g111690), Psy1 (Solyc03g031860), and Psy2 (Solyc02g081330) genes, which are the ripening genes of tomatoes. Tomatoes injected with Agrobacterium containing the CRISPR-Cas9 expression vector were further cultured for 5 days and used to check gene editing efficiency. As a result of the target gene sequence analysis by the next generation sequencing method, gene editing efficiency was calculated, and the efficient target location was selected for the PL and Psy genes. CONCLUSION: Therefore, this study was aimed to establish target sgRNA data that could have higher efficiency of the CRISPR-Cas9 system to obtain the delayed ripening phenotype of tomato. The developed method and sgRNA information is expected to be utilized in the development of various crops to manage its ripening processes.

CRISPR-Cas9 시스템을 활용한 제초제 저항성 토마토 개발 전략

김의연 ( Euyeon Kim ),박효선 ( Hyosun Park ),양소희 ( Sohee Yang ),구연종 ( Yeonjong Koo ) 한국환경농학회 2020 한국환경농학회 학술대회집 Vol.2020 No.-

The development of herbicide-tolerant plants provided a convenient farming method. The five most popular herbicides are divided into glyphosate,sulfonylureas/imidazolinones, glufosinate, norflurasone and oxyfluorfen depending on the reaction target by their mode of action and these herbicides inhibit 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), acetohydroxyacid synthase (AHAS), glutamine synthetase (GS), Phytoene desaturase (PDS) and protoporphyrinogen oxidase (PPO), respectively. Inhibition of EPSPS and AHAS reduces the biosynthesis of phenolic and branched amino acids, respectively, and inhibition of GS and PDS enhances the production of reactive oxygen species and induces plant necrosis. Inhibition of PPO decreases chlorophyll biosynthesis and inhibits plant photosynthesis. These herbicides induce plant death by interacting with their target proteins and the development of herbicide resistant plants is based on the discovery of mutant proteins insensitive to these herbicides. We reviewed the development of herbicide-tolerant plants and derived target amino acids for the production of herbicide-tolerant proteins using the CRISPR/Cas9 system in tomatoes.

Agrobacterium을 이용한 토마토 떡잎에서 CRISPR-Cas9 시스템의 임시발현 시 토마토 떡잎 발달 단계에 따른 유전자교정 효율 변화

김의연 ( Euyeon Kim ),양소희 ( So Hee Yang ),박효선 ( Hyosun Park ),구연종 ( Yeonjong Koo ) 한국환경농학회 2021 한국환경농학회지 Vol.40 No.3

BACKGROUND: Before generating transgenic plant using the CRISPR-Cas9 system, the efficiency test of sgRNAs is recommended to reduce the time and effort for plant transformation and regeneration process. The efficiency of the sgRNA can be measured through the transient expression of sgRNA and Cas9 gene in tomato cotyledon; however, we found that the calculated efficiency showed a large variation. It is necessary to increase the precision of the experiment to obtain reliable sgRNA efficiency data from transient expression. METHODS AND RESULTS: The cotyledon of 11^th, 15^th, 19^th, and 23^rd-day-old tomato (Solanum lycopersicum cv. Micro-Tom) were used for expressing CRISPR-Cas9 transiently. The agrobacterium harboring sgRNA for targeting ALS2 gene of tomato was injected through the stomata of leaf adaxial side and the genomic DNA was extracted in 5 days after injection. The target gene edition was identified by amplifying DNA fragment of target region and analyzing with Illumina sequencing method. The target gene editing efficiency was calculated by counting base deletion and insertion events from total target sequence read. CONCLUSION: The CRISPR-Cas9 editing efficiency varied with tomato cotyledon age. The highest efficiency was observed at the 19-day-old cotyledons. Both the median and mean were the highest at this stage and the sample variability was also minimized. We found that the transgene of CRISPR-Cas9 system was strongly correlated with plant leaf development and suggested the optimum cotyledon leaf age for Agrobacterium-mediated transfection in tomato.

돈분 액비의 아산화질소 발생 저감 효과 검정

이평호 ( Pyeong Ho Lee ),백지현 ( Ji Hyeon Baek ),구연종 ( Yeonjong Koo ) 한국환경농학회 2023 한국환경농학회지 Vol.42 No.4

This study focused on nitrous oxide, a major greenhouse gas produced in agricultural settings through bacterial nitrogen oxidation in aerobic soil. Nitrogen fertilizer in farmland is identified as a primary source of nitrous oxide. The importance of reducing excess nitrogen in soil to mitigate nitrous oxide production is well-known. The study investigated the use of liquefied pig manure as an alternative to urea fertilizer in conventional agriculture. Results showed a more than two-fold reduction in nitrous oxide emissions in pepper cultivation areas with liquefied pig manure compared to that with urea fertilizer. The population of Nitrosospira, a nitrous oxide-producing bacterium, decreased by over 10% with liquefied pig manure. Additionally, nirK and nosZ, which are related to the denitrification process, significantly increased in the urea fertilizer group, whereas levels in the liquefied pig manure group resembled those with no nitrogen treatment. In conclusion, the experiment confirmed that liquefied pig manure can serve as an eco-friendly nitrogen fertilizer, significantly reducing nitrous oxide production, a major contributor to the atmospheric greenhouse effect.

등검은말벌과 꿀벌의 장내 세균 군집 비교

김의연 ( Euyeon Kim ),서정원 ( Jeongwon Seo ),양소희 ( So Hee Yang ),김인선 ( In-seon Kim ),구연종 ( Yeonjong Koo ) 한국환경농학회 2018 한국환경농학회지 Vol.37 No.2

한국에서 말벌은 양봉 산업에 큰 영향을 미치는 종이다. 특히 비교적 최근에 전국적으로 번식하는 말벌 종인 등검은 말벌 (Vespa velutina nigrithorax)은 작은 체구에도 불구하고 높은 사냥 능력으로 꿀벌을 공격하여 양봉 업계에 큰 피해를 주고 있다. 그러나 이 새로운 침입 종에 대한 연구는 아직 시작단계에 있으며, 본 연구에서는 등검은말벌의 장내 박테리아 군집을 조사하고 이 결과를 꿀벌의 장내 박테리아 군집과 비교하여 등검은말벌의 방제를 위한 자료로 이용하고자 했다. 아시아 말벌과 꿀벌 장의 박테리아 게놈 DNA를 수집하여 16S rDNA를 증폭시켜 가변 부위인 V3, V4 염기서열을 판독 하였다. 계통별 분석결과 목 (order) 수준에서 Flavobacteriales는 등검은말벌에서 가장 우점인 박테리아였고, Aeromonadales와 Pseudomonadales가 그 뒤를 이었다. Flavobacteriaes는 꿀벌의 박테리아 군집과 비교했을 때 등검은말벌에서 증가하였으며, Aeromonadales와 Pseudomonadales는 우점종이지만 꿀벌에 비해 낮은 점유율을 보이는 등 두 개체 간의 차이를 확인할 수 있었다. 한편 이번 연구를 통해 이전에 동정되지 않은 등검은말벌 장 박테리아의 16S rDNA염기 서열을 분석하였고, 이들 박테리아는 Thalassomonas, Caedobacter, Vampirovibrio, Alkaliphilus 및 Calothrix 속으로 분류되었다. BACKGROUND: The Asian hornet (Vespa velutina nigrithorax), a wasp species, has attacked honey bee populations and affected the beekeeping industry in Korea over the past 15 years. However, little research has been done with this invasive species. In this study, we investigated the intestine bacterial microbiota of Asian hornets and honey bees to design an attractive trap for Asian hornets. METHODS AND RESULTS: Genomic DNAs isolated from the intestine microorganisms of Asian hornets and honey bees were utilized to amplify bacterial 16S rDNA for the comparative sequence analysis. The next generation sequencing analysis identified that the orders Flavobacteriales as the most abundant intestinal microorganisms in Asian hornets, showing a clear difference compared to honey bees in which Aeromonadales are dominant. We also report five newly identified 16S rDNA sequences of Asian hornet intestinal bacteria. According to the sequence blast search, these five bacteria belong to the genera Thalassomonas, Caedobacter, Vampirovibrio, Alkaliphilus and Calothrix. CONCLUSION: While Asian hornets and honey bees show similar intestine bacterial diversity, the relative ratio of bacterial populations is different. providing useful information to design pest control agents specifically targeting Asian hornets.

농업환경에서 메탄 발생 경로와 메타노젠의 다양성

문수빈,백지현,구연종 전남대학교 농업과학기술연구소 2022 농업생명과학연구동향 Vol.60 No.-

Methane is a potent greenhouse gas that is produced naturally through various bioogical processes, includin the decomposition of organic matter. Two common sources of methane production are cow dung and paddy soil. Cow dung is a rich source of organic matter and is widely used as a fertilizer and fuel. Methane production from cow dung occurs in the rumen of cows durin the digestion process, where the bacteria present in the rumen break down the organic matter and produce methane as a by-product. The major methanogens are members of the Archaea domain, specifically the genus Methan-obrevibacter. These methanogens are known to produce methane by breaking down the complex organic matter in the cow's stomach. Paddy soil is the soil used for cultivating paddy rice, which is flooded with water during the growing season. The flooded conditions create anaerobic conditions, which promote the growth of methanogenic bacteria is the soil. These bcteria brea down the organic matter in the soil and produce methane as a by-product, which is released into the atmosphere through the rice plant roots and the soil surface. The major methanogens are also member of the Archaea domain, but they belong to a different genus called Methanosaeta. Methanosaeta are known to produce methane by metabolizing acetate, which is produced by other microorganisms in the soil. In this report, we compare the methane production of two different agricultural methane sources n terms of bacterial diversity and biochemical pathways for methane production.