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Shinyoung Lee

Publications and source records attributed to Shinyoung Lee.

8 recordsLinked to original sources

Molecular genetics using T-DNA in rice.

Now that sequencing of the rice genome is nearly completed, functional analysis of its large number of genes is the next challenge. Because rice is easy to transform, T-DNA has been used successfully to generate insertional mutant lines. Collectively, several laboratories throughout the world have established at least 200,000 T-DNA insertional lines. Some of those carry the GUS or GFP reporters for either gene or enhancer traps. Others are activation tagging lines for gain-of-function mutagenesis when T-DNA is inserted in the intergenic region. A forward genetic approach showed limited success because of somaclonal variations induced during tissue culture. To utilize these resources more efficiently, tagged lines have been produced for reverse genetics approaches. DNA pools of the T-DNA-tagged lines have been prepared for polymerase chain reaction (PCR) screening of insertional mutants in a given gene. Appropriate T-DNA insertion sites are determined by sequencing the region flanking the T-DNA. This information is then used to make databases that are shared with the scientific community. International efforts on seed amplification and maintenance are needed to exploit these valuable materials efficiently.

DNA, Bacterial↗

Type I MADS-box genes have experienced faster birth-and-death evolution than type II MADS-box genes in angiosperms.

Plant MADS-box genes form a large gene family for transcription factors and are involved in various aspects of developmental processes, including flower development. They are known to be subject to birth-and-death evolution, but the detailed features of this mode of evolution remain unclear. To have a deeper insight into the evolutionary pattern of this gene family, we enumerated all available functional and nonfunctional (pseudogene) MADS-box genes from the Arabidopsis and rice genomes. Plant MADS-box genes can be classified into types I and II genes on the basis of phylogenetic analysis. Conducting extensive homology search and phylogenetic analysis, we found 64 presumed functional and 37 nonfunctional type I genes and 43 presumed functional and 4 nonfunctional type II genes in Arabidopsis. We also found 24 presumed functional and 6 nonfunctional type I genes and 47 presumed functional and 1 nonfunctional type II genes in rice. Our phylogenetic analysis indicated there were at least about four to eight type I genes and approximately 15-20 type II genes in the most recent common ancestor of Arabidopsis and rice. It has also been suggested that type I genes have experienced a higher rate of birth-and-death evolution than type II genes in angiosperms. Furthermore, the higher rate of birth-and-death evolution in type I genes appeared partly due to a higher frequency of segmental gene duplication and weaker purifying selection in type I than in type II genes.

Arabidopsis↗

OsEIN2 is a positive component in ethylene signaling in rice.

EIN2 is a central signal transducer in the ethylene-signaling pathway, and a unique membrane-anchored protein. By screening a cDNA library, we have isolated a cDNA clone (OsEIN2) that encodes the rice EIN2 homolog. The full-length ORF clone was obtained by reverse transcriptase-polymerase chain reaction. OsEIN2 shares significant amino acid sequence similarity with Arabidopsis EIN2 (57% similarity and 42% identity). Both the numbers and positions of introns and exons in the OsEIN2 and AtEIN2 coding regions are also conserved. To address whether this structural similarity is indicative of functional conservation of the corresponding proteins, we also generated transgenic lines expressing the antisense construct of OsEIN2. Those plants were stunted and shoot elongation was severely inhibited. Their phenotypes were similar to that found with wild-type rice seedlings that were treated with AgNO3, an ethylene signal inhibitor. In the OsEIN2 antisense plants, the expression levels of two ethylene-responsive genes, SC129 and SC255, were decreased compared with the wild types. These results suggest that OsEIN2 is a positive component of the ethylene-signaling pathway in rice, just as AtEIN2 is in Arabidopsis: Our antisense transgenic plants produced approximately 3.5 times more ethylene than the wild-type plants. Expression analysis of rice ACS and ACO genes showed that the transcript levels of OsACS1 and OsACO1 were elevated in the transgenic plants.

Amino Acid Sequence↗

Functional analyses of the flowering time gene OsMADS50, the putative SUPPRESSOR OF OVEREXPRESSION OF CO 1/AGAMOUS-LIKE 20 (SOC1/AGL20) ortholog in rice.

A late-flowering mutant was isolated from rice T-DNA-tagging lines. T-DNA had been integrated into the K-box region of Oryza sativa MADS50 (OsMADS50), which shares 50.6% amino acid identity with the Arabidopsis MADS-box gene SUPPRESSOR OF OVEREXPRESSION OF CO 1/AGAMOUS-LIKE 20 (SOC1/AGL20). While overexpression of OsMADS50 caused extremely early flowering at the callus stage, OsMADS50 RNAi plants exhibited phenotypes of late flowering and an increase in the number of elongated internodes. This confirmed that the phenotypes observed in the knockout (KO) plants are because of the mutation in OsMADS50. RT-PCR analyses of the OsMADS50 KO and ubiquitin (ubi):OsMADS50 plants showed that OsMADS50 is an upstream regulator of OsMADS1, OsMADS14, OsMADS15, OsMADS18, and Hd (Heading date)3a, but works either parallel with or downstream of Hd1 and O. sativa GIGANTEA (OsGI). These results suggest that OsMADS50 is an important flowering activator that controls various floral regulators in rice.

Base Sequence↗

Mental health attitudes among Caucasian-American and Korean counseling students.

Attitudinal differences among 104 Korean and 107 Caucasian-American counseling students was investigated using the Mental Health Values Questionnaire (MHVQ). Caucasian-American students reported significantly greater attitudinal scores on the negative traits, untrustworthiness, and unconventional reality subscales. In contrast, the Korean sample reported higher attitudinal scores on the achievement and affective control subscales. Implications for cross-cultural mental health education and practice are discussed.

Adult↗

The OsFOR1 gene encodes a polygalacturonase-inhibiting protein (PGIP) that regulates floral organ number in rice.

We have isolated a cDNA clone, OsFOR1, from the immature panicles of rice. The OsFOR1 (Oryza sativa floral organ regulator 1) gene encodes a protein that contains a leucine-rich repeat (LRR) domain. This domain comprises 10 tandem repeats of a canonical 24-amino acid LRR sequence. The structure and the number of LRRs for OsFOR1 are similar to those of polygalacturonase-inhibiting proteins (PGIPs) from various other plant species. Moreover, the OsFOR1 recombinant protein, when fused to maltose-binding protein (MBP), shows PGIP activity against the Aspergillus niger polygalacturonase. OsFOR1 is highly expressed in the calli and immature and mature panicles, while detectable at only low levels in seedling roots and mature stems. In situ hybridization experiments showed the transcripts of OsFOR1 are present in young spikelet primordia and in almost all of the young floral organs. Transgenic approaches were used to study in vivo functioning. Antisense expression of OsFOR1 resulted in an increase in the numbers of floral organs, including the stamen, carpel, palea/lemma, stigma, and lodicule. OsFOR1 transcript was not detected in the frizzy panicle mutant, which is defective in its spikelet formation but normal in inflorescence-meristem initiation and maintenance. Therefore, we suggest that OsFOR1 plays a role in the formation and/or maintenance of floral organ primordia.

Amino Acid Sequence↗

Systematic reverse genetic screening of T-DNA tagged genes in rice for functional genomic analyses: MADS-box genes as a test case.

We have generated 47 DNA pools and 235 subpools from 21,049 T-DNA insertion lines of rice. DNA pools of 500-1,000 lines were adequate for screening a T-DNA insertion within a 2-kb region. To examine the efficacy of the DNA pools, we selected MADS-box genes, which play an important role in controlling various aspects of plant development. A total of 34 MIKC-type MADS-box genes have now been identified from rice sequence databases. Our PCR screening for T-DNA insertions within 12 MADS-box genes resulted in the identification of five insertions in four different genes. These DNA pools will be valuable when isolating T-DNA insertional mutants in various rice genes. The DNA pool screening service and the mutant seeds are available upon request to genean@postech.ac.kr.

DNA, Bacterial↗

Characterization of tobacco MADS-box genes involved in floral initiation.

MADS-box genes encode regulatory factors that are involved at various stages in plant development. These genes function not only during early floral meristem identity, but also when the fate of floral organ primordia is determined in a later step. Here, we screened a floral bud cDNA library to isolate a tobacco MADS-box gene, NtMADS4, using the rice MADS-box gene, OsMADS1, as a probe. We previously reported that OsMADS1 plays a critical role in flower development in rice. Ectopic expression of NtMADS4 caused phenotypes of extremely early flowering as well as dwarfism. Plant MADS proteins have a K domain that mediates the formation of dimers. This dimerization appears to be an essential step for a functional protein complex. NtMADS11 was isolated as an interacting partner of NtMADS4 by yeast two-hybrid screening. The latter was included in the AGAMOUS-like 2 (AGL2) family whereas the former was categorized in the SQUAMOSA (SQUA) family. While the transcript of NtMADS4 was detectable only in reproductive organs, that of NtMADS11 was seen in both reproductive and vegetative organs. Expression levels were high for both genes during early developmental stages. Ectopic expression of NtMADS11 and OsMADS14 was able to rescue the floral organ defects seen in the strong ap1-1 mutant. Roles of NtMADS4 and NtMADS11 in the floral initiation are discussed.

Amino Acid Sequence↗