Development and Identification of Transgenic Rice Lines with Abiotic Stress Tolerance by using a Full-length Overexpressor Gene Hunting System

Abstract

This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. ABSTRACT The latest report on the draft genome of Brassica rapa sequence has been published. To elucidate the functions of these genes and to efficiently search for agriculturally useful genes, a Full-length cDNA Over-eXpressor (FOX) gene hunting system was used. The FOX library from Chinese cabbage was introduced into rice via Agrobacterium-mediated transformation. Approximately 1,150 FOX-rice lines were generated. Genomic PCR analysis indicated that the average length of FL-cDNAs introduced into individual lines was 900~1,200 bp. Basic Local Alignment System Tool (BLAST) analysis of the FL-cDNA genes revealed that 35.5% have unknown function. Most of the randomly selected transgenic rice lines showed overexpression (92%) of these genes relative to the wild-type Gopum. Moreover, 94% of the 850 transgenic rice lines were moderately tolerant (slightly yellow) to cold and 9 lines were tolerant (seedlings were light green). Morphological evaluation of the transgenic rice lines showed minimal phenotypic alteration (12%). Approximately 25.1% and 22% of the plants were significantly ahead in the days to heading and had elevated chlorophyll content, respectively. Other agronomic traits such as filled grains, number of tiller, panicle length, and culm and plant height were relatively less variable among the transgenic lines. These results provide a resource for defining genes that are associated with tolerance in transgenic rice lines.

Overexpression of BrCIPK1 Gene Enhances Abiotic Stress Tolerance by Increasing Proline Biosynthesis in Rice

“…From the large FOX rice lines we developed and screened for abiotic stresses tolerance published earlier (Abdula et al 2013), BrCIPK1 was selected which showed tolerance to multiple abiotic stresses. The BrCIPK1 full-length complementary DNA (cDNA) was ligated into the pSB11 vector.…”

“…Indeed, in spite of having very similar amino sequences, these proteins respond differently to stress and regulate different signaling pathways (Pandey et al 2004). To facilitate the study of agronomically important genes, we developed 1500 FOX rice lines and evaluated them against abiotic stresses (Abdula et al 2013). Of these, BrCIPK1 plants showed consistently enhanced tolerance.…”

Enhancement of Rice Resistance to Bacterial Blight by Overexpressing BrCP3 Gene of Brassica rapa

“…To facilitate overexpression, integration of the transgenes was constructed under the control of CaMV 35S and ubiquitin-1 promoter and NOS terminator (Abdula et al 2013). The whole construct was transformed into japonica rice cv.…”

“…Overexpression of this insert gene was constructed under the control of ubiquitin -1 promoter. The binary vector also carries hygromycin phosphotransferase (HPT) gene as a selectable marker (Abdula et al 2013). …”

Gene expression and SNP identification related to leaf angle traits using a genome-wide association study in rice (Oryza sativaL.)

“…그 밖에 qLTG-3-1 (Fujino et al 2014 (Kump and Javomik 1996;Cho et al 1998). 추 출한 DNA는 λDNA (100 ng/ul)와 함께 1% agarose gel에서 확 인하였고 20 ng/ul로 정량하여 실험에 사용하였다 (Abdula et al 2013). 벼 유전자원의 resequencing은 HiSeq 2500 (Illumina) NGS (Next Generation DNA Sequencer)을 이용하여 short read sequence를 공주대학교 박용진 교수팀에서 생산하였다 .…”