해충저항성 Bt벼와 낙동벼의 미꾸리(Misgurnus anguillicaudatus) 와 잉어(Cyprinus carpio)에 대한 급성독성시험을 실시한 결과 48시간 및 96시간-LC50은 1,000mg/L 이상으로 나타났다. 48시간 및 96시간 무영향농도(NOEC)는 1,000mg/L이었다. 급성독성 시험기간 중 해충저항성 Bt벼와 낙동벼간의 pH, DO, 수온, 체중 및 전장에 대한 유의적인 결과는 나타나지 않았다.

본 연구의 목적은 방화곤충에 의해 유전자변형 작물로부터 비변형작물로의 유전자 이동이 일어날 수 있는가를 조사하기 위한 프로토콜을 개발하는 것이다. 이를 위해 제초제저항성 형질전환 콩과 재배콩인 태광콩, 그리고 G. soja 시험구에서 발생하는 곤충상을 sweeping법으로 조사하였으며, 이들 시험구에서 화분매개 가능성을 가진 방화벌의 시간대별 출현빈도 및 부위별 화분 부착율을 조사하였다. 또한 채집한 서양종꿀벌로부터 부착된 화분을 이용, 핵산을 추출하고 PCR 증폭 가능성 여부를 분석하였다. 그 결과, 유전자변형 콩과 비변형 콩 사이의 곤충상에는 시기별로 유의성 있는 차이는 없는 것으로 조사되었다. 개화기 콩 꽃에서 서양종꿀벌의 출현빈도는 정오에 가장 높은 것으로 나타났다. 이들이 수확한 화분으로부터 핵산을 추출하여 PCR 을 수행하였을 때 lectin 유전자가 증폭된 것으로 보아 이들 화분은 콩으로부터 유래한 것임을 확인하였다.

Natural and artificially induced mutants have provided valuable resources for plant genetic studies and crop improvement. Some variations induced in the process of plant transformation have often been observed in regenerated plants. In this study, we investigated the insertion number of transgene and the flanking sequences of T-DNA in tall-induced line BP23, which was unexpectedly gained in the process of transformation of insect-resistant rice with cryBP1 gene, and also analyzed the whole-genome sequencing by using the NGS technologies to gain a better understanding of the sequence and structural changes between tall line or natural cultivar and rice reference. than others, was confirmed with two copies of foreign gene insertion, which was inserted in one genomic site facing each other between the position 2,430,152~2,430,151 of rice chromosome 12 without any deletion of genomic sequences. Sequencing analysis also revealed that 18bp-unknown sequences were added in the 5′ insertion site of T-DNA. This position in rice genome was confirmed with none of expressed gene sites. By the NGS analysis, we detected 86560 SNPs and 1091/1472 large insertion/deletion (indel) sites (100bp) between BP23 and rice reference, and 84743 SNPs and 1094/1451 large indels between natural cultivar Nagdong and rice reference. The possible mechanisms for the gene mutation, the developmental and tissue expression of the taller height in BP23 line may need to be scrutinized a few more.

Four transgenic rice lines harboring insect-resistant gene cry3A showed ideal field performances characterized by high considerable resistance to rice water weevil (Lissorhoptrus oryzophilus Kuschel). In this study, we estimated the insert number of foreign genes, and analyzed the flanking sequences of T-DNA in rice genome. As a result, The T-DNA of Btt12R 3-1-1-1 line was inserted in exon region of rice chromosome 10 and Btt12R 6-1-1-1 line was inserted in two copies of foreign gene. Btt12R 9-1-1-1 line was analyzed at only left border flanking sequence. The T-DNA of Btt12R 13-1-1-1 line was inserted one copy of foreign gene between position 24,516,607~24,516,636 of rice chromosome 5 and 30bp known genomic sequences were deleted. The Btt12R 13-1-1-1 line confirmed to be inserted in intergenic region having not any expressed gene and no any deletion/addition of T-DNA sequence. From these results, we demonstrated that the molecular data of rice water weevil resistant Bt rice could be acceptable to conduct the biosafety and environment risk assessment for GM crop commercialization

The rice water weevil (RWW), Lissorhoptrus oryzophilus are major pests of aquatic rice plant in Korea as well as throughout the country. Larvae of RWW sucking the nourishment on roots, causes a stunted root system and reduces grain yields. To prevent these damages, we constructed various plant expression vectors, which were harbored by insecticidal genes, cryBP1 and cryIIIa, and fused with the actin promoter and/or the modified RCg2 root-preferential promoters for expressing the insect-toxic genes in leaves and roots. A cryBP1 was cloned from Bacillus popilliae, producing crystal toxin against Japanese beetle, and CryIIIa was modified from the δ-endotoxin gene of Bacillus thuringiensis ssp. tenebrionis, encoding the coleoptera-specific toxin. The vectors containing the insecticidal genes were transferred into Oryza sativa japonica cultivar, Nakdong, by Agrobacterium -mediated transformation method. Several independent transgenic lines were selected by Southern blotting and Western blotting, confirming that cryBP1 and cryIIIa genes were stably integrated into the plant genomes and were expressed in transgenic plants. Upon insect bioassay using RWW, the mortality of insect larvae on cryBP1 and cryIIIa transgenic rice lines recorded up to 41% and 34%, respectively. These results suggested that the transgenic lines can be used to develop Coleoptera-resistant cultivars and could be valuable for later application in crop breeding for insect resistance.

Over 7 individual rice (Oryza sativa L.) plants per a line were sowed and sampled by pooled sampling method for genomic DNA extraction. The 5,400 flanking sequence tags (FSTs) were analysed by adaptor PCR and direct sequencing. FST analysis showed that the intragenic FSTs, the intergenic FSTs, and the original insertional sequences including hot spot covered 48.1% (2,597), 25.6% (1,383), and 25% (1,350), respectively. The 2,597 intragenic FSTs were used for genotyping to determine whether they are heterozygous or homozygous, and 1,393 core lines were selected. Among them, 422 knockout genes were distributed on chromosome 3, while 56 - 157 intragenic FSTs scattered on other chromosomes. Among 1,393 FSTs, known genes such as transcription factor covered 59.4% (827), while unknown genes such as expressed protein covered 40.6% (566). RT-PCR indicated that some core lines had no expression or decreased expression level in their knockout genes. It means that core lines are very useful knockout lines for functional genomic studies.

1. Ds 삽입변이체 3,000계통으로부터 제초제 저항성 1,874계통을 선발하고 농업적 주요 특성으로서 출수일수, 간장, 수수, 수장, 엽장 등 5가지 형질에 대하여 조사한 바, 조사된 5가지 형질에 대하여 원품종인 동진벼에 비하여 매우 다양한 변이폭을 보여주었다. 2. 농업적 유용성과 관련된 수장이 길고, 조기출수, 수수가 많은 변이체 뿐만 아니라 형태학적 변이를 보이는 twin seedling, dwarf, early heading, strip albino, liguleless 등 변이체가 다수 발견됨으로서 육종적 이용 및 유전자 기능해석을 위한 유용한 집단으로서 유용성을 보여주었다. 3. 서던분석 결과, 벼 게놈상에서 Ds는 평균 2 copy로 전이되었으며 조직부위별로 GUS의 발현을 조사한 결과 잎, 뿌리 및 화기관등에서 약 3.9%가 발현되었다. 이 삽입변이체에서 나타난 다양한 변이형질의 주요 농업적 특성과 GUS 발현의 재현성을 위해 다음 세대의 전개를 통한 후대분석이 필요하다.