Background : Cudrania tricuspidata Bureau is a widely used medicinal perennial woody plant. Obtaining information about the genetic diversity of plant populations is highly important for conservation and germplasm utilization. In this study, we developed single nucleotide polymorphism (SNP) markers derived from chloroplast genomic sequences to identify distinct Korean-specific ecotypes of C. tricuspidata via amplification refractory mutation system (ARMS)-PCR analyses. We performed molecular authentication of twelve C. tricuspidata ecotypes from different regions using DNA sequences in the chloroplast TrnL-F intergenic region. Methods and Results : SNPs were identified based on the results of nucleotide sequence for the intergenic region of TrnL-TrnF gene (chloroplast). Molecular markers were designed for those SNPs with additional mutations on the second base from SNPs for amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). HRM pattern analyses were performed using the Mx3005P QPCR System (Agilent Technologies, CA, USA). Conclusion : We collected 12 individual lines of C. tricuspidata from various region in South Korea and China. Based on the nucleotide sequence in the trnL-trnF intergenic region of these lines, six SNPs and a deletion of 12 bps were identified and 12 individual lines were able to be grouped in one Korean ecotype and two different ecotypes of chinese lines, chinese line 1 and 2. The SNP markers developed in this study are useful for rapidly identifying these specific C. tricuspidata ecotypes collected from different regions.
Background : The P. ginseng breeding line G07006, was selected for salt tolerance through salinity screening of mature leaves at the NIHHS of the RDA in 2014-2016. However, it is difficult to maintain a genetically stable breeding line of cross-pollinating crop in the field. Therefore molecular marker required to identify and maintain breeding line G07006. Methods and Results : DNA was extracted following the CTAB DNA extraction protocol (Doyle and Doyle, 1987) with modifications. A pair-end (PE) library was constructed and sequenced using an Illumina MiSeq platform by Lab Genomics, Inc. (Seongnam, Korea). Approximately 4.0 Gb of sequencing data were obtained, and de novo assembled by a CLC genome assembler(v. beta 4.6, CLC Inc., Rarhus, Denmark). The complete chloroplast(CP) genome size is 156,356 bp, including two inverted repeats (IRs) of 52,060 bp, separated by the large single-copy (LSC 86,174 bp) and small single-copy (SSC 18,122 bp) regions. This CP genome encodes 114 unigenes (80 protein-coding genes, four rRNA genes, and 30 tRNA genes), in which 18 are duplicated in the IR regions. Conclusion : This complete chloroplast DNA sequence will provide conducive to discriminate line G070006 (salt-tolerant) and further enhancing genetic improvement program of this important medical plant.
1. 본 연구에서는 완두유래의 세포질성 PsAPX1 유전자를 대상으로 산화스트레스 유도성 프로모터를 연결하여 엽록체에 targeting 되는 과발현 운반체를 제작하고 벼에 도입한 결과 형질전환체에서 도입유전자 수가 1~3 copy인 것으로 나타나, 적은 수의 유전자가 안정적으로 도입되었음을 확인하였다. 2. 염, 오존, 자외선, 한발과 같은 다양한 환경스트레스 조건에서 내성이 증진된 우수 계통을 선발하기 위하여 작성된 형질전환 벼 계통들을 대상으로 생물
염색의 농담에 따른 엽록소의 함량과 수량과의 관계를 밝히고자 연녹색잎 표지인자를 가진 Taichung 65(pgl)와 녹색엽인 수원345호 및 두 계통간의 교잡 BC1 F2 에서 농업형질과 엽록체의 특성을 밝히고자 수행한 결과를 요약하면 다음과 같다. 1. 연녹색 및 계통인 T.65(pgl)와 수원345호의 조합 F2 에서 연녹색 잎 개체군은 T.65(pgl)와 차이가 없었으나 녹색잎 개체군의 평균 개체수량은 수원345호에 비하여 월등히 높게 나타났다. 2. T.65(pgl)의 엽록소 함량은 수원345호에 비하여 적었으며, 감소되는 유형은 비슷하였으나 Ca/Cb 비율은 출수후 15일부터 60일까지 오히려 높게 나타났다. 3. 출수 후 T.65(pgl)의 지엽은 수원345호에 비하여 낮은 SPAD 값을 보였지만 2엽, 3엽은 모두 유의한 차이가 없었다. 반면 F2 에서 연녹색잎 개체군의 지엽, 2엽, 3엽에서의 SPAD 값은 녹색잎 개체군에 비하여 모두 낮게 나타났다. 4. 출수후 20일에 지엽의 엽록체를 투과전자현미경으로 검경한 결과 T.65(pgl)에서 osmium granule의 수가 많은 것으로 나타났다.