• The Plant Pathology Journal
• /
• 제38권4호
• /
• pp.296-303
• /
• 2022

The cucurbit powdery mildew (CPM) caused by different fungal species is a major concern for cucurbit crops around the world. In Argentina CPM constitutes the most common and damaging disease for cucurbits, especially for squash crops (Cucurbita moschata). The present study displays initial insights into the knowledge of the disease in western Argentina, including the determination of the prevalent species causing CPM, as well as the evaluation of the resistance of squash cultivars and breeding lines. Fungal colonies were isolated from samples collected in Mendoza province, Argentina. A field trial was also performed to assess the resistance of five squash accessions, including commercial cultivars and breeding lines. The severity of CPM was analyzed and epidemiological models were built based on empirical data. The morphological determinations and analysis with specific molecular markers confirmed Podosphaera xanthi as the prevalent causal agent of CPM in Mendoza. The results od the field trial showed differences in the resistance trait among the squash accessions. The advanced breeding line BL717/1 showed promising results as source of CPM resistance for the future development of open pollinated resistant cultivars, a crucial tool for an integrative control of the disease.

• Journal of Plant Biotechnology
• /
• 제48권1호
• /
• pp.18-25
• /
• 2021

멕시코로부터 유래한 Solanum demissum은 감자 야생종 중의하나로 감자 역병에 대해 저항성을 가지고 있어 감자 육종에서 중요한 재료로 이용되고 있다. S. demissum의 EBN은 4배 체인 감자와 같은 4로 직접적인 교배로 육종에 활용될 수 있다. 본 연구에서는 NGS 기술에 의해 완성된 S. demissum의 엽록체 전장 유전체(cpDNA)와 이를 다른 Solanum종과의 비교를 통해 개발한 분자마커에 대해 보고하였다. S. demissum의 전체 cpDNA의 크기는 155,558 bp였으며 그 구조는 다른 Solanum종과 매우 유사하였다. S. demissum의 cpDNA와 가지과에 속하는 10개 종의 cpDNA 코딩서열을 이용하여 분석한 계통수에서는 S. demissum이 S. hougasii 및 S. stoloniferum과 거의 동일한 유전체 구성을 보였으며, 다음으로 S. berthaultii 및 S. tuberosum과 유연관계가 가까운 것으로 확인되었다. S. demissum과 다른 7종의 Solanum과의 전체 cpDNA 다중 정렬을 통해 S. demissum 특이적인 두 개의 InDel 영역을 구명하였으며 이를 기반으로 최종적으로 PCR을 기반으로 한 두 개의 S. demissum 특이적 마커를 개발하였다. 본 연구의 결과는 Solanum 종들을 대상으로 한 조금 더 세부적인 진화적 그리고 육종적 측면에서의 연구에 기여를 할 수 있을 것이다.

• Journal of Plant Biotechnology
• /
• 제31권1호
• /
• pp.1-6
• /
• 2004

한약재로 사용되는 방풍류는 절단되어 유통되므로 외부 형태적인 특징만으로 구분하기가 어려워 방풍류로 사용되는 방풍, 식방풍, 석방풍, 갯방풍 등 4종에 대해 PCR에 기초한 RAPD 마커를 이용하여 SCAR 마커를 개발하고자 하였다. RAPD 분석결과 밴드의 패턴은 다양하게 나타났으며 다형성의 밴드 수는 총 215개로 전체 밴드수의 98%였다. RAPD 분석에서 각 방풍류를 구별 할 수 있는 특이적인 밴드를 나타내는 primer는 방풍에서 4개의 primer, 식방풍은 6개의 primer, 석방풍은 4개의 primer, 갯방풍은 6개의 primer를 선발하였고, 그 중 특히 primer 425는 4종의 방풍류의 감별에 유용하였고, 이를 이용하여 SCAR마커로 전환하는데 이용하였다. 특이적인 단편을 클로닝하여 염기서열 분석으로 특이 primer를 제작하고 제작된 primer로 방풍류 시료 16개에 적용하였을 때, 국내의 야생에서 주로 자생하는 석방풍은 215 bp, 그리고 국내에서 가장 많이 재배 또는 생산되는 갯방풍은 177 bp와 300 bp에서 뚜렷하게 나타났다. 따라서 갯방풍과 석방풍의 감별 가능성을 제시할 수 있으며 개발된 SCAR 마커를 이용하여 시중에 유통되고 있는 방풍류 건조약재의 감별에 유용한 마커로 활용될 수 있을 것이다.

• The Plant Pathology Journal
• /
• 제34권4호
• /
• pp.308-315
• /
• 2018

Root-knot nematodes 'Meloidogyne spp' are the most destructive group of plant parasitic nematodes causeing serious losses in vegetables crops and this damages worsened when crops grown under greenhouses conditions. In this sutdy, the distribution and characterization of root-knot nematode species collected from the Souss region of Morocco where vegetables crops intensively cultivated were determined by using both morphological and molecular tools. Out of the 110 samples collected from different greenhouses 91 (81.7%) were found to be infested with root-knot nematodes. Thirty-seven populations of root-knot nematodes were morphologically identified based on perineal patterns as well as molecularlly using species-specific primers. The obtained results indicated that Meloidogyne javanica and M. incognita were identified in 86.4% and 13.5% of the total populations, respectively. The lowest incidence of root-knot nematodes (64%) was found in Toussous, whereas the highest frequencies of 100% and 90% were detected in Taddart and Biogra, respectively. As the majority of the samples have been infested with Meloidogyne species; this indicates that there is an urgent need to provide farmers with a proper control strategy.

• Journal of Ecology and Environment
• /
• 제43권3호
• /
• pp.320-331
• /
• 2019

Background: Dioecious plant species having both male and female plants have been investigated regarding sex-related characteristics such as sex ratio, sex-differential resource requirements, and spatial segregation of the sexes. Habitat loss and fragmentation are major threats to the survival of plant populations, but dioecious species are particularly more prone to such habitat degradation than non-dioecious species because of their dimorphic sexual system. We examined the sex-related demographics of two Ilex cornuta populations being different regarding land use history. Methods: During 2016-2017, we examined I. cornuta trees with a basal diameter ${\geq}1.5cm$ in the Yongsu-ri population (YS population) and the Gotjawal Provincial Park population (GP population). Plant sex (male, female, or unsexed) was identified. The tree size (basal diameter and height of the main stem), clonal production (the ramet numbers per genet), and vitality for each clone were measured. The associations between population, sex, tree size, clonal production, and vitality were examined using ANOVAs and contingency table analyses. Finally, point pattern analyses using O-ring statistics were conducted to assess spatial patterns. Results: Upon excluding unsexed trees, the YS population with 74 trees was significantly male-biased (0.66), while the GP population with only 26 trees had a 1:1 sex ratio. In both populations, males and females did not differ in tree size. Although the mean number of ramets differed significantly between populations, females tended to produce more ramets than males. The proportion of weak trees was significantly higher in the YS than in the GP population. Neither population showed evidence of spatial segregation of the sexes. Conclusions: The two populations of dioecious I. cornuta are characterized by the small number of trees and relatively high frequencies of non-reproductive trees. Both indicate that these populations are quite susceptible to environmental and genetic stochasticity. On the other hand, the differences between populations in sex ratio, clonal production, and vitality suggest that conservation efforts for I. cornuta need to be population-specific. In order to help recover and enable this vulnerable species to persist, it is necessary to find ways to enhance their sexual reproduction and simultaneously reduce habitat disturbances due to anthropogenic activities.

• 한국자원식물학회지
• /
• 제26권3호
• /
• pp.397-402
• /
• 2013

Morus Folium (Sang-yeop in Korean) is one of the most important Oriental medicinal plants. In Korea, both M. alba and M. cathayana are regarded as the botanical sources for Morus Folium. In order to discriminate M. alba and M. cathayana from their adulterant, M. tricuspidata, mitochondrial NADH dehydrogenase subunit 7 (nad7) intron 2 region was targeted for molecular analysis with universal primers. DNA polymorphisms, including SNP sites, insertions, and deletions, were detected among these three species sequencing data. Based on these DNA polymorphisms, specific primers were designed for the three species respectively. Multiplex PCR was conducted for molecular authentication of M. alba, M. cathayana, and M. tricuspidata with specific primers. The present results indicate that it is possible to identify Morus Folium from its adulterant using mitochondrial nad7 intron 2 region. The established multiplex-PCR system was proved to be effective for identification of Morus Folium. The results indicate that mitochondrial introns can be used for inter-specific polymorphic study, and the described method can be applied for molecular identification of medicinal materials.

• Journal of Plant Biotechnology
• /
• 제2권2호
• /
• pp.55-60
• /
• 2000

Experiments initiated 30 years ago to obtain selectable markers have led to a series of studies of Trp biosynthesis and anthranilate synthase (AS) the control enzyme using largely plant tissue cultures since they have experimental properties that can be readily exploited. Enzymological and compound feeding studies provided evidence that AS is the control point in the Trp biosynthesis branch and that altering the AS feedback control by the selection of mutants resistant to the Trp analog 5-methyl-tryptophan (5MT) can lead to the overproduction of this important amino acid. Plants regenerated from these Trp overproducing lines of most species also had high free Trp levels but Nicotiana tabaum (tobacco) plants expressed the feedback altered AS only in cultured cells and not in the regenerated plants. further tests by transient and stable expression of the cloned promoter for the naturally occurring tobacco feedback-insensitive AS, denoted ASA2, confirmed the tissue culture specific nature of the expression control. The 5MT caused by the expression of a feedback-insensitive AS from tobacco has been used to select protoplast fusion hybrids with several species since the resistance is expressed dominantly. Recently the ASA2 gene has been used successfully as a selectable marker to select transformed Astragalus sinicus and Glycine max hairy roots induced by Agrobactetium rhizogenes. These results show that the ASA2y-subunit can interact with the y-subunit of another species to form active feedback-insensitive enzyme that may be useful for selecting transformed cells. Plastid DNA transformation of tobacco has also effectively expressed ASA2 in the compartment in which Trp biosynthesis is localized in the cell.

• The Plant Pathology Journal
• /
• 제36권3호
• /
• pp.204-217
• /
• 2020

In nature, plants are always under the threat of pests and diseases. Pathogenic bacteria are one of the major pathogen types to cause diseases in diverse plants, resulting in negative effects on plant growth and crop yield. Chemical bactericides and antibiotics have been used as major approaches for controlling bacterial plant diseases in the field or greenhouse. However, the appearance of resistant bacteria to common antibiotics and bactericides as well as their potential negative effects on environment and human health demands bacteriologists to develop alternative control agents. Bacteriophages, the viruses that can infect and kill only target bacteria very specifically, have been demonstrated as potential agents, which may have no negative effects on environment and human health. Many bacteriophages have been isolated against diverse plant-pathogenic bacteria, and many studies have shown to efficiently manage the disease development in both controlled and open conditions such as greenhouse and field. Moreover, the specificity of bacteriophages to certain bacterial species has been applied to develop detection tools for the diagnosis of plant-pathogenic bacteria. In this paper, we summarize the promising results from greenhouse or field experiments with bacteriophages to manage diseases caused by plant-pathogenic bacteria. In addition, we summarize the usage of bacteriophages for the specific detection of plant-pathogenic bacteria.

• Journal of Plant Biotechnology
• /
• 제45권1호
• /
• pp.9-16
• /
• 2018

DNA 바코딩 기술은 다양한 약용식물 종들의 기원을 확인하기 위해 폭넓게 이용되고 있는 연구방법이다. 그러나, 시중에 판매되고 있는 유사 식물종을 재료로 사용한 상품이나 혼재되어 있는 상품에서 확인하고자 하는 약용식물을 선별 가능한 실질적인 기술의 개발은 아직 많이 미흡한 실정이다. 최근에는 보다 신속하고 정확도가 높은 기술을 개발하고자 DNA barcoding (Bar) 기술과 high-resolution melting (HRM) curve pattern 분석기술을 혼합한 Bar-HRM 분석기술을 이용한 연구가 진행 중에 있다. 본 리뷰논문에서는 국제적인 시장에서 다양한 기원의 약용식물 판별에 실질적으로 적용 가능한 Bar-HRM 기술의 최근의 발전 과정과 그 이용에 대해서 정리하였다. 다양한 연구들을 통해서 일부 성공적인 결과들이 보고되고 있지만, 제한된 DNA 바코드 및 단일염기다형성(Single Nucleotide Polymorphism, SNP) 등 아직 해결되어야 할 과제들이 많다. 특히, 핵 내 바코드로는 ribosomal DNA의 internal transcribed sequence (ITS)단편 이외에는 보고된 사례가 한건도 없었다. 또한, 약용식물을 끓는 물로 추출하여 가공한 약탕, 잼, 젤리, 쥬스 등의 제품은 DNA 단편이 분해되어 분리가 안 되는 경우에는 DNA바코딩 기술을 적용하기가 곤란한 것으로 알려져 있으나 비교적 짧은 DNA단편이 요구되는 Bar-HRM 분석기술을 이용하여 일부 성공한 보고도 있어 향후 그 응용사례가 증가할 것으로 전망된다.

• Molecules and Cells
• /
• 제26권3호
• /
• pp.250-257
• /
• 2008

Microsatellites or simple sequence repeats (SSR) are widely distributed in eukaryotic genomes and are informative genetic markers. Despite many advantages of SSR markers such as a high degree of allelic polymorphisms, co-dominant inheritance, multi-allelism, and genome-wide coverage in various plant species, they also have shortcomings such as low polymorphic rates between genetically close lines, especially in Capsicum annuum. We developed an alternative technique to SSR by normalizing and alternating anchored primers in random amplified microsatellite polymorphisms (RAMP). This technique, designated reverse random amplified microsatellite polymorphism (rRAMP), allows the detection of nucleotide variation in the 3' region flanking an SSR using normalized anchored and random primer combinations. The reproducibility and frequency of polymorphic loci in rRAMP was vigorously enhanced by translocation of the 5' anchor of repeat sequences to the 3' end position and selective use of moderate arbitrary primers. In our study, the PCR banding pattern of rRAMP was highly dependent on the frequency of repeat motifs and primer combinations with random primers. Linkage analysis showed that rRAMP markers were well scattered on an intra-specific pepper map. Based on these results, we suggest that this technique is useful for studying genetic diversity, molecular fingerprinting, and rapidly constructing molecular maps for diverse plant species.