• Journal of Ginseng Research
• /
• 제35권1호
• /
• pp.52-59
• /
• 2011

Recently, new ginseng cultivars having superior agricultural traits have been developed in Korea. For newly developed plant cultivars, the identification of distinctiveness is very important factors not only in plant cultivar management but also in breeding programs. Thus, eighty-five inter simple sequence repeat (ISSR) primers were applied to detect polymorphisms among six major Korean ginseng cultivars and two foreign ginsengs. A total of 197 polymorphic bands with an average 5.8 polymorphic bands and 2.9 banding patterns per assay unit across six Korean ginseng cultivars and foreign ginsengs from 236 amplified ISSR loci with an average 6.9 loci per assay unit were generated by 34 out of 85 ISSR primers. Three species of Panax ginseng including the Korean ginseng cultivars, P. quinquefolius, and P. notoginseng, could be readily discriminated using most tested primers. UBC-821, UBC-868, and UBC-878 generated polymorphic bands among the six Korean ginseng cultivars, and could distinguish them from foreign ginsengs. Sequence characterized amplified region (SCAR) marker system was introduced in order to increase the reproducibility of the polymorphism. One SCAR marker, PgI821C650, was successfully converted from the randomly amplified polymorphism by UBC-821. It showed the expected dominant polymorphism among ginseng samples. In addition, the specific polymorphism for Sunwon was generated by treating Taq I restriction enzyme to polymerase chain reaction products of PgI821C650. These results will serve as useful DNA markers for identification of Korean ginseng, especially Sunwon cultivar, seed management, and molecular breeding program supplemented with marker-assisted selection.

• 원예과학기술지
• /
• 제29권6호
• /
• pp.600-609
• /
• 2011

Allium속에 포함된 6종의 122점을 수집하고, 이 종들의 유전적 관계는 ISSR 마커를 이용하여 확인하였다. 형태적 분석은 6개의 양적 형질을 측정하고 1개의 질적 형질은 수치화하였다. SSR 분석은 17개의 primer를 사용하여 총 370개의 다형성 밴드를 얻었다. 형태적 특성 분석은 유전적 거리로 구분할 경우 3개의 그룹으로 분류되었으나, 부분적으로 몇 몇 종들은 분류에 어려움이 있었다. ISSR 결과를 바탕으로 Allium 속의 군집분석은 5개의 그룹으로 분리되었다. 형태적 분석과 SSR분석 간의 상관 관계는 유의성이 매우 낮았다(r = 0.036). 따라서 본 연구에서 개발한 ISSR 마커는 달래와 산달래의 분류와 교배 육종에 유용하게 이용될 수 있을 것이다.

• 생명과학회지
• /
• 제17권1호
• /
• pp.24-30
• /
• 2007

꽃치자(Gardenia jasminoides var. radicans)와 열매치자(C. jasminoides for. grandiflora)는 Gardenia속으로는 우리나라에 두 분류군밖에 없으며 열매치자는 약용, 식용, 꽃치자는 방향제로 쓰인다. 그런데 꽃이 지고 나면 형태적으로 구분이 거의 되지 않는다 ISSR분석으로 이들 종의 유전적 다양도와 집단구조를 실시하였다. 88개의 DNA 분절에서 한 분류군에만 나타나는 특이밴드가 탐지되었다. 한국의 세 야생 집단은 분리되어 있고 패치 분포를 보이지만 재배종에 비해 높은 유전적 다양성을 유지하친 있었다. 열매치자가 꽃치자보다 유전적 다양성이 높았으며 ISSR 마커로 이들 분류군이 잘 분리되었다. 또한 야생 집단이 재배 집단보다 다양성이 약간 높으나 유의성은 없었다. 이는 재배화과정에서 유전적 다양성의 일부 상실이 있었으나 인위적인 채취와 식재로 야생 집단과 재배 집단의 유전적 교류가 존재하였음을 시사한다.

• Plant Breeding and Biotechnology
• /
• 제2권3호
• /
• pp.224-230
• /
• 2014

Amplified fragment length polymorphism (AFLP) is a molecular marker technique based on DNA and is extremely useful in detection of high polymorphism between closely related genotypes like Korean wheat cultivars. Six sequence characterized amplified regions (SCARs) have been developed from inter simple sequence repeat (ISSR) analysis which enabled the identification and differentiation of 13 Korean wheat cultivars from the other cultivars. We used six combinations of primer sets in our AFLP analysis for developing additional cultivar-specific markers in Korean wheat. Fifty-eight of the AFLP bands were isolated from EA-ACG/MA-CAC, EA-AGC/MA-CTG and EA-AGG/MA-CTA primer combinations. Of which 40 bands were selected to design SCAR primer pairs for Korean wheat cultivar identification. Three of 58 amplified primer pairs, KWSM006, KWSM007 and JkSP, enabled wheat cultivar identification. Consequently, 23 of 32 Korean wheat cultivars were classified by eight SCAR marker sets.

• 식물조직배양학회지
• /
• 제28권3호
• /
• pp.129-134
• /
• 2001

Winter bud explants from 15 individual angelica tree (Aralia elata) were cultured in vitro to find out optimal conditions for somatic embryo induction as well as plant regeneration. Calli are induced and grown on MS medium supplemented with 1.0 mg/L 2,4-D for 4 weeks and subcultured on a half-strength MS medium without phytohormones to induce somatic embryos. Inter-simple sequence repeat (I-SSR) markers were analyzed with total DNAs extracted from the trees. Genotype effects on somatic embryo induction were examined by cluster analysis. Callus induction rate varied from 58.5 to 100% among the genotypes. Somatic embryo induction rate also greatly varied from 0 to 100% among the genotypes. There was a significant difference in somatic embryo induction rate even among the individual trees that showed close genetic relationships each other. This suggested that somatic embryo induction rate in Aralia elata be influenced by a few major specific genes rather than whole genomic similarity among individual trees. Four individuals of Ulneong-7, Cheju-1, Shingu and China, which are recalcitrant to somatic embryo induction, turned out to have a close genetic relationship, suggesting that both physiological and genetic factors affect somatic embryo induction. The results suggest that genotype selection be the most important factor to achieve an efficient propagation, although cultural optimization through medium and explant manipulation may also play crucial roles in somatic embryogensis as well as plant regeneration of these species.

• Plant Biotechnology Reports
• /
• 제2권4호
• /
• pp.239-243
• /
• 2008

Simmondsia chinensis (Link) Schneider, a multipurpose and monogeneric dioecious shrub from arid zones, has emerged as a cash crop all over the globe. Its seed propagation poses severe problems due to its male-biased population: the male:female ratio is 5:1. Investigations have been carried out to generate a sex-specific Inter-simple sequence repeat (ISSR) marker for the early detection of male and female plants. Of the 42 primers analysed with a bulk sample of pooled male DNA and a bulk sample of pooled female DNA, only one primer, UBC-807, produced a unique ~1,200 base-pair fragment in the male DNA. To validate this observation, this primer was re-tested with individual male and female samples from eight cultivars. A similar unique ~1,200 bp fragment was present in the male individuals of all eight cultivars and completely absent in the female individuals tested. This is the first report of the use of ISSR markers to ascertain sex in physiologically mature S. chinensis plants.

• Journal of Microbiology and Biotechnology
• /
• 제20권4호
• /
• pp.683-692
• /
• 2010

Morphologically, nine different slow-growing protoclones were screened from regenerated protoplasts of heterokaryotic Agaricus bisporus. As such, the present study is the first report on differentiating homo- and heterokaryotic protoclones using random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers. Among 80 primers tested, the seven ISSR and seven RAPD primers selected for the analysis generated a total of 94 ISSR and 52 RAPD fragments, respectively. The ISSR fingerprinting also detected more polymorphic loci (38.29%) than the RAPD fingerprinting (34.61%). A principal coordinate analysis (PCA) was employed to evaluate the resolving power of the markers as regards differentiating protoclones. As a result, the mean polymorphism information content (PIC) for each marker system (i.e., 0.787 for RAPD and 0.916 for ISSR) suggested that ISSR is more effective for determining polymorphisms. The dendrograms constructed using RAPD, ISSR, and an integrated RAPD and ISSR marker system were highly correlated with one another as revealed by a high Mantel correlation (r= 0.98). The pairwise similarity index values also ranged from 0.64 to 0.95 (RAPD), 0.67 to 0.98 (ISSR), and 0.67 to 0.98 (RAPD and ISSR), whereas the mean similarity index values of 0.82, 0.81, and 0.84 were obtained for the RAPD, ISSR, and combined data, respectively. As there was a good correspondence between the RAPD and ISSR similarity matrices, ISSR would appear to be an effective alternative to RAPD in the genetic diversity assessment and accurate differentiation of homo- and heterokaryotic protoclones of A. bisporus.

• 식물분류학회지
• /
• 제47권1호
• /
• pp.11-22
• /
• 2017

Prangos fedtschenkoi (Regel et Schmalh.) Korovin (Apiaceae) is an endemic species for mountainous Middle Asia, which is both a rare and useful plant. Organic extractions from this species are being used in pharmaceutics and cosmetology. In recent years, P. fedtschenkoi distribution area has considerably decreased, presumably, resulting from human activities such as agriculture, construction works, overgrazing and collection from wild for pharmaceutic purposes. Six populations were found in Uzbekistan and their genetic divergence and differentiation were studied with 10 inter-simple sequence repeat (ISSR) markers, selected out of 101. Totally 166 amplified ISSR fragments (loci) were revealed, of which 164 were polymorphic. Relatively moderate level of polymorphism was found at population level with polymorphic bands ranging from 27.71% to 47.59%. Mean P = 39.05%, $N_a=1.40$, $N_e=1.25$, S.I. = 0.21, and $H_e=0.14$ were revealed for all loci across six populations. AMOVA showed higher variation among populations (62%) than within them (38%). The Bayesian model determined 5 clusters, or genetic groups. The posteriori distribution of the Theta II estimator detected full model identifying high inbreeding, intensified by low gene flow (Nm = 0.3954). Mantel test confined population 6 as distinct cluster corresponding to geographic remoteness (R = 0.5137, $p{\leq}0.005$). Results were used as the bases for developing conserve measures to restore populations.

• 한국자원식물학회지
• /
• 제28권6호
• /
• pp.697-703
• /
• 2015

Twenty apple germplasm accessions from the Korean Genebank were successfully cryopreserved using two-step freezing to back up genetic resources maintained by field collections. This study examined the morphological and genetic stability of cryopreserved dormant apple buds that were stored in liquid nitrogen, and then rewarmed and regrown. Whole plants were regenerated directly from dormant buds through budding without an intermediary callus phase. The cryopreserved buds produced high levels of shoot formation (76.2-100%), similar to those of noncryopreserved buds (91.3-100%), with no observed differences between cryopreserved and noncryopreserved materials. Three of the twenty cryopreserved apple germplasm accessions were used to assess morphological and genetic stability. No differences in morphological characteristics including shoot length, leaf shape, leaf width/length ratio, and root length were observed between controls (fresh control and noncryopreserved) and cryopreserved plantlets. The genetic stability of regenerants (before and after cryopreservation) was investigated using inter simple sequence repeat (ISSR) markers. The ISSR markers produced 253 bands using four primers, ISSR 810, SSR 835, ISSR 864, and ISSR 899. These markers showed monomorphic banding patterns and revealed no polymorphism between the mother plant and regenerants before and after cryopreservation, suggesting that cryopreservation using two-step freezing does not affect the genetic stability of apple germplasm. These results show that two-step freezing cryopreservation is a practical method for long-term storage of apple germplasms.

• Journal of Plant Biotechnology
• /
• 제47권4호
• /
• pp.298-308
• /
• 2020

Genetic diversity among Thaumatococcus daniellii populations in the southwestern region of Nigeria were assessed using morphometric and molecular markers to determine the population structure and existing genetic relationship for its improvement, conservation and sustainable utilisation. Populations from five locations in each of the six states were used for the study. Morphometric data were collected on folia characters and analysed for variability. Genome DNA was isolated from the plant leaf and amplified by polymerase chain reaction with inter-simple sequence repeat markers (ISSR) to determine the allelic polymorphism, marker effectiveness and genetic relationship of the population. The results showed significant variations in petiole length and leaf dimensions of the populations within and across the states. These morphometric traits are the major parameters that delimit the populations and they correlated significantly at P≤0.05. Analysis of the electrophoregram showed that the ISSR markers are effective for the diversity study. A total of 136 loci were amplified with an average of 7.16 loci per marker, 63.2% of the loci were polymorphic. The Principal Coordinate Analysis revealed that seven factors accounted for 81.6% of the variation and the dendrogram separated the populations into two major groups at a genetic distance of 10 (about 90% similarity) with sub-groups and clusters. Most populations within the state had a high degree of similarity, nonetheless, strong genetic relationship exists among populations from different states. The close relationship between populations across the states suggests a common progenitor, which are likely separated by ecological or geographical isolation mechanisms.