• Journal of Plant Biotechnology
• /
• 제40권1호
• /
• pp.9-17
• /
• 2013

Ran is a small GTP-binding protein that binds and subsequently hydrolyzes GTP. The functions of Ran in nuclear transport and mitotic progression are well conserved in plants and animals. In animal cells, stress treatments cause Ran relocalization and slowing of nuclear transport, but the role of Ran proteins in plant cells exposed to stress is still unclear. We have therefore compared Ran genes from three EST libraries construed from different cell types of sweetpotato and the distribution pattern of Ran ESTs differed according to cell type. We further characterized two IbRan genes. IbRan1 is a specific EST to the suspension cells and leaf libraries, and IbRan2 is specific EST to the root library. IbRan1 showed 94.6 % identity with IbRan2 at the amino acid level, but the C-terminal region of IbRan1 differed from that of IbRan2. These two genes showed tissue-specific differential regulation in wounded tissues. Chilling stress induced a similar expression pattern in both IbRan genes in the leaves and petioles, but they were differently regulated in the roots. Hydrogen peroxide treatment highly stimulated IbRan2 mRNA expression in the leaves and petioles, but had no significant effect on IbRan1 gene expression. These results showed that the transcription of these two IbRan genes responds differentially to abiotic stresses and that they are subjected to tissue-specific regulation. Plant Ran-type small G-proteins are a multigenic family, and the characterization of each Ran genes under various environmental stresses will contribute toward our understanding of the distinctive function of each plant Ran isoform.

• The Plant Pathology Journal
• /
• 제37권2호
• /
• pp.99-114
• /
• 2021

Tan spot of wheat, caused by Pyrenophora tritici-repentis (Ptr), results in a yield loss through chlorosis and necrosis of healthy leaf tissue. The major objective of this study was to compare gene expression in resistant and susceptible wheat cultivars after infection with Ptr ToxA-producing race 2 and direct infiltration with Ptr ToxA proteins. Greenhouse experiments included exposure of the wheat cultivars to pathogen inoculum or direct infiltration of leaf tissue with Ptr-ToxA protein isolate. Samples from the experiments were subjected to RNA sequencing. Results showed that ToxA RNA sequences were first detected in samples collected eight hours after treatments indicating that upon Ptr contact with wheat tissue, Ptr started expressing ToxA. The resistant wheat cultivar, in response to Ptr inoculum, expressed genes associated with plant resistance responses that were not expressed in the susceptible cultivar; genes of interest included five chitinases, eight transporters, five pathogen-detecting receptors, and multiple classes of signaling factors. Resistant and susceptible wheat cultivars therefore differed in their response in the expression of genes that encode chitinases, transporters, wall-associated kinases, permeases, and wound-induced proteins, among others. Plants exposed to Ptr inoculum expressed transcription factors, kinases, receptors, and peroxidases, which are not expressed as highly in the control samples or samples infiltrated with ToxA. Several of the differentially expressed genes between cultivars were found in the Ptr resistance QTLs on chromosomes 1A, 2D, 3B, and 5A. Future studies should elucidate the specific roles these genes play in the wheat response to Ptr.

• 한국식물생명공학회:학술대회논문집
• /
• 한국식물생명공학회 2002년도 추계학술대회
• /
• pp.33-40
• /
• 2002

We, Tong Yang Moolsan Co. Ltd. (TYM) set up the mass-production system for virus-free seed garlic via tissue culture technique. TYM's tissue culture technique is called as 'Multiple shoot propagation technique'. This technique can lead mass propagation of genetically homogeneous seed garlic in a short period because of its highly proliferation rate of in vitro shoots ($15^{10}$ /year). TYM researchers applied the technique to some selected garlic cultivars with superior characteristics and carried out field test of productivity in the inside and outside of the country for several years. According to the yearly results of field test with virus-free seed garlic, we ascertained that virus-free seed garlic can produce the highly yield increase (max. above $50\%$) and also can enhance the product quality. Consequently, we estimated that TYM's seed garlic will contribute to farmers with increase of income and can elevate the national position of garlic market in the world for its competitive power of technical and production cost.

• Journal of Plant Biology
• /
• 제22권1_2호
• /
• pp.5-14
• /
• 1979

In several leguminous plants such as acasia, arrowroot and bushclover, growth rate and contents of nitrogen, phosphorus and potassium in the tissues and the variation in the culture media were determined. In water cultrue which was free of added nutrients, nitrogen was found to be largely in the form of nitrate(NO3-N). This NO3-N is believed to be the result of nitrification from NH4-N which was apparently released form the plants. From the studies of organ culture with root segments, the amount of nitrogen released and absorbed was found to be proportional to the amount added to the mediuim. Especially, in the N-plot, the amount of nitrogen absorbed by the tissue reached more than 90% of the amount supplied to the medium already in early stage. On the contrary, in the amount free plot, the amount of nitrogen released from the tissue was lower than the minimum level in the N-plot. The amount of total N and P in the cultured tissue was found to be influenced by the amount of nitrogen addedin the medium. However, the amount of K in the tissue was not related to the nitrogen level in the medium, but rather it was influenced by the amount of added potassium. These findings present little difference in the metabolic pattern among the three species plants studied, and suggest that the woody leguminous plants have some common features in tehir metabolic pattern.

• Journal of Plant Biotechnology
• /
• 제37권4호
• /
• pp.357-369
• /
• 2010

Cymbidium is horticulturally important and has been one of the most commercially successful orchid plants as well as cut flowers around the world including Korea. Up to now, a huge number of elite Cymbidium cultivars have been released on the commercial market via cross-hybridization, mutation and polyploidization breeding techniques. To investigate on breeding system in Cymbidium, we inquired the brief history and techniques of breeding and the current status on Cymbidium breeding in Korea. Also, the general propagation process of elite Cymbidium lines via tissue culture should be presented. However, the slow process of conventional breeding and the lack of useful genes in Cymbidium species delays the introduction of new cultivars to the commercial market. To solve these limitations, efficient regeneration and genetic transformation systems should be established in the improvement of Cymbidium breeding program. During the last several decades, some progress has been made in tissue culture and genetic transformation in Cymbidium species. We review the recent status of tissue culture and genetic transformation systems in Cymbidium plants.

• Journal of Plant Biotechnology
• /
• 제39권1호
• /
• pp.13-22
• /
• 2012

Plant regeneration has been optimized increasingly by organogenesis and somatic embryogenesis using a range of explants with tissue culture improvements focusing on factors, such as the age of the explant, genotype, media supplements and $Agrobacterium$ co-cultivation. The production of haploids and doubled haploids using microspores has accelerated the production of homozygous lines in Brassicaceae crop plants. Somatic cell fusion has facilitated the development of interspecific and intergeneric hybrids in sexually incompatible species of $Brassica$. Crop improvement using somaclonal variation has also been achieved. Transformation technologies are being exploited routinely to elucidate the gene function and contribute to the development of novel enhanced crops. The $Agrobacterium$-mediated transformation is the most widely used approach for the introduction of transgenes into Brassicaceae, and $in$ $vitro$ regeneration is a key factor in developing an efficient transformation method in plants. Although many other Brassicaceae are used as model species for improving plant regeneration and transformation systems, this paper focuses on the recent technologies used to regenerate the most important Brassicaceae crop plants.

• Journal of Plant Biotechnology
• /
• 제33권4호
• /
• pp.263-270
• /
• 2006

Pongamia pinnata Pierre is a tree legume, having potential in production of raw material for biodiesel. A protocol for in wk propagation of this plant was standardized using seedling explants. Growth regulators (GR) including gibberellic acid $(GA_3),\;N^6-benzylaminopurine(BA)$, thidiazuron (TDZ), and Adenine sulphate (Ads) were tested for optimum germination of seeds. Removal of seed coat prior to germination, controlled fungal growth partially but enhanced bacterial growth. Antibiotic cefotaxime was ineffective in controlling bacterial contamination. Seedling derived nodal explants and cotyledon nodes with attached cotyledons were excised and cultured for induction of shoots. Optimum sprouting and multiplication of shoot buds were obtained in MS medium supplemented with $8.88{\mu}M$ BA. These buds differentiated and rooted on medium devoid of GR. Optimum growth of Pongamia seedling was obtained in cotton plugged culture vessels. Reculturing of the cotyledon node explants produced more shoots from the same site. This process of removing shoots and reculturing of cotyledon node was followed for eight passages yielding 4 to 8 shoots in each cycle. The shoots (75%) rooted on half strength MS basal medium supplemented with 0.22% charcoal. All plants survived on transfer to soil. This is the first report on in vitro regeneration of Pongamia pinnata. This report demonstrates the possibility of coupling more than one parameter in single experiment to hasten the process of standardization. The process of cycling the nodal explant repeatedly for production of large number of shoots from single meristem may find application in genetic transformation experiments wherein meristems are used for transformation.

• Molecules and Cells
• /
• 제44권10호
• /
• pp.746-757
• /
• 2021

Plant somatic cells can be reprogrammed into a pluripotent cell mass, called callus, which can be subsequently used for de novo shoot regeneration through a two-step in vitro tissue culture method. MET1-dependent CG methylation has been implicated in plant regeneration in Arabidopsis, because the met1-3 mutant exhibits increased shoot regeneration compared with the wild-type. To understand the role of MET1 in de novo shoot regeneration, we compared the genome-wide DNA methylomes and transcriptomes of wildtype and met1-3 callus and leaf. The CG methylation patterns were largely unchanged during leaf-to-callus transition, suggesting that the altered regeneration phenotype of met1-3 was caused by the constitutively hypomethylated genes, independent of the tissue type. In particular, MET1-dependent CG methylation was observed at the blue light receptor genes, CRYPTOCHROME 1 (CRY1) and CRY2, which reduced their expression. Coexpression network analysis revealed that the CRY1 gene was closely linked to cytokinin signaling genes. Consistently, functional enrichment analysis of differentially expressed genes in met1-3 showed that gene ontology terms related to light and hormone signaling were overrepresented. Overall, our findings indicate that MET1-dependent repression of light and cytokinin signaling influences plant regeneration capacity and shoot identity establishment.

• Journal of Plant Biotechnology
• /
• 제49권3호
• /
• pp.230-239
• /
• 2022

Holostemma annulare (Family Asclepiadaceae) is an invaluable vulnerable medicinal plant; the root tubers are used in Ayurveda medicine and by folk healers to treat various ailments. In this study, Schenk and Hildebrandt medium fortified with the cytokinins 6-benzyl adenine, kinetin, and auxins, including indole 3-butyric acid, indole 3-acetic acid, α-naphthaleneacetic acid, and 2,4-dichlorophenoxyacetic acid, were checked for their efficiency on root tuber induction from different explants. Adventitious root tubers were more successfully induced from in vitro leaf segments and shoots when cultured in Schenk and Hildebrandt medium supplemented with 0.5 mg/l of α-naphthaleneacetic acid. In addition, preliminary phytochemical analysis of in vitro root tubers and identification of different secondary metabolites were conducted. Thin layer chromatography and high performance thin layer chromatography analysis of the crude methanolic extracts of the in vitro root tuber identified the presence of lupeol, a bioactive triterpene. Adventitious root tuber induction offers a novel method for the in vitro production of bioactive metabolites that can be scaled up by bioreactors, thus ensuring the conservation and sustainable utilization of H. annulare. The study warrants further scale-up production and pharmacological investigation that can be extended for pharmaceutical needs.

• Journal of Plant Biotechnology
• /
• 제50권
• /
• pp.108-114
• /
• 2023

In vitro conservation is one of the most effective strategies for rare plant protection, especially for orchid species. To maximize the success rates of in vitro explant establishment (stage I) in conservation programs, the application of tissue culture additives such as Plant Preservative Mixture^TM (PPM^TM) should be emphasized. In this study, we used Dendrobium thyrsiflorum Rchb.f. (1875) seeds and seedlings as a model for the evaluation of PPM^TM's phytotoxicity in the meristematic tissues of epiphytic orchids. PPM^TM had no observable inhibitory effect on protocorm, shoot, or root development when it was supplemented at 0.1%. PPM^TM supplementation caused adverse effects on D. thyrsiflorum explants at concentrations > 0.2%. At high concentrations, young in vitro seedlings showed damage, especially at the root tissue level. Based on this model, supplementation of 0.1-0.2% PPM^TM to culture media was successfully implemented to establish in vitro cultures of other rare orchid species in our conservation program.