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Development of a Female-associated SCAR Marker in Schisandra nigra Max. (Schisandra nigra Max.에서 암그루에 연관된 SCAR 마커의 개발)

  • Han, Hyo Shim;Jung, Jae Sung
    • Journal of Life Science
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    • v.31 no.6
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    • pp.537-542
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    • 2021
  • Schisandra nigra Max., a dioecious plant native to Jeju Island in Korea, is cultivated on a small scale for fruit production. As fruit-producing female individuals are generally considered to be more valuable than male, early identification of plant sex at the seedling stage is important. In this study, a sequence-characterized amplified region (SCAR) marker associated with a female-specific region in the genome of S. nigra was investigated. Of 120 randomly amplified polymorphic DNA (RAPD) primers, one primer (OPB-03) consistently amplified a 749 bp band in female plants. The female-specific PCR product was isolated and cloned, and the nucleotide sequences were then determined. Southern hybridization performed using the female-specific fragment as a probe produced positive signals only in genomic DNA from the female plants. This result revealed that the 749 bp segment of DNA was present in the genome of female plants but absent in the genome of male plants. A SCAR primer pair was designed based on the RAPD marker to amplify a 436 bp fragment in the genomic DNA of female plants. This primer pair amplified the expected size of DNA fragment in female plants and four monoecious individuals collected from a natural population. The SCAR marker identified in this study can be used to distinguish female-flowering individuals at the seedling stage.

Paleomagnetism, Stratigraphy and Geologic Structure of the Tertiary Pohang and Changgi Basins; K-Ar Ages for the Volcanic Rocks (포항(浦項) 및 장기분지(盆地)에 대한 고지자기(古地磁氣), 층서(層序) 및 구조연구(構造硏究); 화산암류(火山岩類)의 K-Ar 연대(年代))

  • Lee, Hyun Koo;Moon, Hi-Soo;Min, Kyung Duck;Kim, In-Soo;Yun, Hyesu;Itaya, Tetsumaru
    • Economic and Environmental Geology
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    • v.25 no.3
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    • pp.337-349
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    • 1992
  • The Tertiary basins in Korea have widely been studied by numerous researchers producing individual results in sedimentology, paleontology, stratigraphy, volcanic petrology and structural geology, but interdisciplinary studies, inter-basin analysis and basin-forming process have not been carried out yet. Major work of this study is to elucidate evidences obtained from different parts of a basin as well as different Tertiary basins (Pohang, Changgi, Eoil, Haseo and Ulsan basins) in order to build up the correlation between the basins, and an overall picture of the basin architecture and evolution in Korea. According to the paleontologic evidences the geologic age of the Pohang marine basin is dated to be late Lower Miocence to Middle Miocene, whereas other non-marine basins are older as being either Early Miocene or Oligocene(Lee, 1975, 1978: Bong, 1984: Chun, 1982: Choi et al., 1984: Yun et al., 1990: Yoon, 1982). However, detailed ages of the Tertiary sediments, and their correlations in a basin and between basins are still controversial, since the basins are separated from each other, sedimentary sequence is disturbed and intruded by voncanic rocks, and non-marine sediments are not fossiliferous to be correlated. Therefore, in this work radiometric, magnetostratigraphic, and biostratigraphic data was integrated for the refinement of chronostratigraphy and synopsis of stratigraphy of Tertiary basins of Korea. A total of 21 samples including 10 basaltic, 2 porphyritic, and 9 andesitic rocks from 4 basins were collected for the K-Ar dating of whole rock method. The obtained age can be grouped as follows: $14.8{\pm}0.4{\sim}15.2{\pm}0.4Ma$, $19.9{\pm}0.5{\sim}22.1{\pm}0.7Ma$, $18.0{\pm}1.1{\sim}20.4+0.5Ma$, and $14.6{\pm}0.7{\sim}21.1{\pm}0.5Ma$. Stratigraphically they mostly fall into the range of Lower Miocene to Mid Miocene. The oldest volcanic rock recorded is a basalt (911213-6) with the age of $22.05{\pm}0.67Ma$ near Sangjeong-ri in the Changgi (or Janggi) basin and presumed to be formed in the Early Miocene, when Changgi Conglomerate began to deposit. The youngest one (911214-9) is a basalt of $14.64{\pm}0.66Ma$ in the Haseo basin. This means the intrusive and extrusive rocks are not a product of sudden voncanic activity of short duration as previously accepted but of successive processes lasting relatively long period of 8 or 9 Ma. The radiometric age of the volcanic rocks is not randomly distributed but varies systematically with basins and localities. It becomes generlly younger to the south, namely from the Changgi basin to the Haseo basin. The rocks in the Changgi basin are dated to be from $19.92{\pm}0.47$ to $22.05{\pm}0.67Ma$. With exception of only one locality in the Geumgwangdong they all formed before 20 Ma B.P. The Eoil basalt by Tateiwa in the Eoil basin are dated to be from $20.44{\pm}0.47$ to $18.35{\pm}0.62Ma$ and they are younger than those in the Changgi basin by 2~4 Ma. Specifically, basaltic rocks in the sedimentary and voncanic sequences of the Eoil basin can be well compared to the sequence of associated sedimentary rocks. Generally they become younger to the stratigraphically upper part. Among the basin, the Haseo basin is characterized by the youngest volcanic rocks. The basalt (911214-7) which crops out in Jeongja-ri, Gangdong-myon, Ulsan-gun is $16.22{\pm}0.75Ma$ and the other one (911214-9) in coastal area, Jujon-dong, Ulsan is $14.64{\pm}0.66Ma$ old. The radiometric data are positively collaborated with the results of paleomagnetic study, pull-apart basin model and East Sea spreading theory. Especially, the successively changing age of Eoil basalts are in accordance with successively changing degree of rotation. In detail, following results are discussed. Firstly, the porphyritic rocks previously known as Cretaceous basement (911213-2, 911214-1) show the age of $43.73{\pm}1.05$$49.58{\pm}1.13Ma$(Eocene) confirms the results of Jin et al. (1988). This means sequential volcanic activity from Cretaceous up to Lower Tertiary. Secondly, intrusive andesitic rocks in the Pohang basin, which are dated to be $21.8{\pm}2.8Ma$ (Jin et al., 1988) are found out to be 15 Ma old in coincindence with the age of host strata of 16.5 Ma. Thirdly, The Quaternary basalt (911213-5 and 911213-6) of Tateiwa(1924) is not homogeneous regarding formation age and petrological characteristics. The basalt in the Changgi basin show the age of $19.92{\pm}0.47$ and $22.05{\pm}0.67$ (Miocene). The basalt (911213-8) in Sangjond-ri, which intruded Nultaeri Trachytic Tuff is dated to be $20.55{\pm}0.50Ma$, which means Changgi Group is older than this age. The Yeonil Basalt, which Tateiwa described as Quaternary one shows different age ranging from Lower Miocene to Upper Miocene(cf. Jin et al., 1988: sample no. 93-33: $10.20{\pm}0.30Ma$). Therefore, the Yeonil Quarterary basalt should be revised and divided into different geologic epochs. Fourthly, Yeonil basalt of Tateiwa (1926) in the Eoil basin is correlated to the Yeonil basalt in the Changgi basin. Yoon (1989) intergrated both basalts as Eoil basaltic andesitic volcanic rocks or Eoil basalt (Yoon et al., 1991), and placed uppermost unit of the Changgi Group. As mentioned above the so-called Quarternary basalt in the Eoil basin are not extruded or intruaed simultaneously, but differentiatedly (14 Ma~25 Ma) so that they can not be classified as one unit. Fifthly, the Yongdong-ri formation of the Pomgogri Group is intruded by the Eoil basalt (911214-3) of 18.35~0.62 Ma age. Therefore, the deposition of the Pomgogri Group is completed before this age. Referring petrological characteristics, occurences, paleomagnetic data, and relationship to other Eoil basalts, it is most provable that this basalt is younger than two others. That means the Pomgogri Group is underlain by the Changgi Group. Sixthly, mineral composition of the basalts and andesitic rocks from the 4 basins show different ground mass and phenocryst. In volcanic rocks in the Pohang basin, phenocrysts are pyroxene and a small amount of biotite. Those of the Changgi basin is predominant by Labradorite, in the Eoil by bytownite-anorthite and a small amount pyroxene.

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Genes of Wild Rice (Oryza grandiglumis) Induced by Wounding and Yeast Extract (상처와 효모추출물 처리조건에서 유발되는 야생벼 유전자 스크린)

  • Shin, Sang-Hyun;Im, Hyun-Hee;Lee, Jai-Heon;Kim, Doh-Hoon;Chung, Won-Bok;Kang, Kyung-Ho;Cho, Sung-Ki;Shin, Jeong-Sheop;Chung, Young-Soo
    • Journal of Life Science
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    • v.14 no.4
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    • pp.650-656
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    • 2004
  • Oryza grandiglumis (CCDD, 2n=48), one of the wild rice species, has been known to possess fungal-,bacterial-, and insect-resistance against sheath blight, rice blast, bacterial leaf blight and brown plant hopper (Nilaparvata lugens). To rapidly isolate differentially expressed genes responding to fungal and wounding stress, wounding and yeast extract were treated to O. grandiglumis for 24 hrs. Suppression subtractive hybridization (SSH) method was used to obtain differentially expressed genes from yeast extract and wounding treated plants. Seven hundreds and seventy six clones were obtained by subcloning PCR product, and colony array and screening were carried out using radio-isotope labeled cDNA probes prepared from the wounding and yeast extract treated plants. One hundred and fifteen colonies were confirmed as true positive ones. Average insert size of the clones were ranged from 400 bp to 700 bp and all the inserts were sequenced. To decide the identity of those clones, sequences were analyzed by sequence homology via GenBank database. The homology search result showed that 68 clones were matched to the genes with known function; 16 were related to primary metabolism, 5 to plant retrotransposons, 5 to defense related metallothionein-like genes. In addition to that, others were matched to various genes with known function in amino acid synthesis and processing, membrane transport, and signal transduction, so on. In northern blot analysis, induced expressions of ogwfi-161, ogwfi-646, ogwfi-663, and ogwfi-695 by wounding and yeast extract treatments were confirmed. The result indicates that SSH method is very efficient for rapid screening of differentially expressed genes.

Cloning, cSNP Identification, and Genotyping of Pig Complement Factor B(CFB) Gene Located on the SLA Class III Region (SLA Class III 영역의 돼지 Complement Factor B(CFB) 유전자의 Cloning, cSNP 동정 및 유전자형 분석)

  • Kim, Jae-Hwan;Lim, Hyun-Tae;Seo, Bo-Yeong;Zhong, Tao;Yoo, Chae-Kyoung;Jung, Eun-Ji;Jeon, Jin-Tae
    • Journal of Animal Science and Technology
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    • v.50 no.6
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    • pp.753-762
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    • 2008
  • The primers for RT-PCR and RACE-PCR were designed by aligning the pig genomic sequence and the human complement factor B(CFB) coding sequence(CDS) from the GenBank. Each PCR product was amplified in pig cDNA and sequencing was carried out. The CDS length of pig CFB gene was determined to be 2298 bp. In addition, the pig CDS was more longer than human and mouse orthologs because of insertion and deletion. The identities of porcine nucleotide sequences with those of human and mice were 84% and 80%, and the identities of amino acids were 79% to 77%, respectively. Three complement control protein(CCP) domains, one Von Willebrand factor A(VWFA) domain and a serine protease domain, that are revealed typically in mammals, were found in the pig CFB gene. Based on the CDSs determined, the primers were designed in intron regions for amplification of entire length of exons. In amplification and direct sequencing with genomic DNAs of six pig breeds, three cSNPs(coding single nucleotide polymorphisms) were identified and verified as missense mutations. Using the Multiplex-ARMS method, we genotyped and verified the mutations identified from direct sequencing. To demonstrate recrudescence, we performed both direct sequencing and Multiplex-ARMS with two randomly selected DNA samples. The genotype of each sample exhibited the same results using both methods. Therefore, three cSNPs were identified from pig CFB gene and that can be used for haplotype analysis of the swine leukocyte antigen(SLA) class III region. Moreover, the results indicate that the Multiplex-ARMS method should be powerful for genotyping of genes in the SLA region.

Identification of a Potexvirus in Korean Garlic Plants (한국 마늘 Potexvirus의 cDNA 유전자 분리 및 분포에 관한 연구)

  • Song, Jong-Tae;Choi, Jin-Nam;Song, Sang-Ik;Lee, Jong-Seob;Choi, Yang-Do
    • Applied Biological Chemistry
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    • v.38 no.1
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    • pp.55-62
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    • 1995
  • To understand the molecular structure of Korean garlic viruses, cDNA cloning of virus genomic RNA was attempted. Virus particles were isolated from virus-infected garlic leaves and a cDNA library was constructed from garlic virus RNA. One of these clones, S81, selected by random sequencing has been identified as a member of potexvirus group other than potyvirus and carlavirus. The clone is 873 bp long contains most of the coat protein (CP) coding region and 3'-noncoding region including poly(A) tail. A putative polyadenylation signal sequence (AAUAAA) and the hexanucleotide motif (ACUUAA), a replicational cis-acting element conserved in the 3'-noncoding region of potexvirus RNAs are noticed. The clone S81 shows about 30-40% identity in both nucleotide and amino acid sequences with CPs of potexviruses. The genome size of the virus was analysed to be 7.46 knt by Northern blot analysis, which was longer than those of other potexviruses. The open reading frame encoding CP was expressed as a fusion protein (S81CP) in Escherichia coli and the recombinant protein was purified by immobilized metal binding affinity chromatography. Polyclonal antibody was raised against S81CP in rabbit to examine the occurrence of garlic potexvirus in Korean garlic plants by immunoblot analysis. Two virus protein bands of Mr 27,000 and 29,000 from garlic leaf extract of various cultivars reacted with the antibody. It was shown that Mr 27,000 band might not be a degradation product of Mr 29,000 band, suggesting that two types of potexvirus different in size of coat protein could exist in Korean garlic plants.

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