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  • Research ArticleMarch 16, 2023

    0 607 202

    Comparative proteome profiling in the storage root of sweet potato during curing-mediated wound healing

    Ho Yong Shin ·Chang Yoon Ji ·Ho Soo Kim ·Jung-Sung Chung ·Sung Hwan Choi ·Sang-Soo Kwak · Yun-Hee Kim·Jeung Joo Lee

    J Plant Biotechnol (2023) 50:001-010

    https://doi.org/10.5010/JPB.2023.50.001.001

    Abstract

    Abstract : Sweet potato (Ipomoea batatas L. Lam) is an economically important root crop and a valuable source of nutrients, processed foods, animal feeds, and pigment materials. However, during post-harvest storage, storage roots of sweet potatoes are susceptible to decay caused by various microorganisms and diseases. Post-harvest curing is the most effective means of healing wounds and preventing spoilage by microorganisms during storage. In this study, we aimed to identify proteins involved in the molecular mechanisms related to curing and study proteomic changes during the post-curing storage period. For this purpose, changes in protein spots were analyzed through 2D-electrophoresis after treatment at 33°C (curing) and 15°C (control) for three days, followed by a storage period of eight weeks. As a result, we observed 31 differentially expressed protein spots between curing and control groups, among which 15 were identified. Among the identified proteins, the expression level of ‘alpha-amylase (spot 1)’ increased only after the curing treatment, whereas the expression levels of ‘probable aldo-keto reductase 2-like (spot 3)’ and ‘hypothetical protein CHGG_01724 (spot 4)’ increased in both the curing and control groups. However, the expression level of ‘sporamin A (spot 10)’ decreased in both the curing and control treatments. In the control treatment, the expression level of ‘enolase (spot 14)’ increased, but the expression levels of ‘chain A of actinidin-E-64 complex+ (spot 19)’, ‘ascorbate peroxidase (spot 22)’, and several ‘sporamin proteins (spot 20, 21, 23, 24, 27, 29, 30, and 31)’ decreased. These results are expected to help identify proteins related to the curing process in sweet potato storage roots, understand the mechanisms related to disease resistance during post-harvest storage, and derive candidate genes to develop new varieties with improved low-temperature storage capabilities in the future.

  • Research ArticleMarch 27, 2023

    0 1070 219

    Somatic embryogenesis induction in four cassava landraces in East Java, Indonesia

    Slameto ・Indri Fariroh ・Budi Kriswanto ・Didik Pudji Restanto ・Kacung Hariyono

    J Plant Biotechnol (2023) 50:011-018

    https://doi.org/10.5010/JPB.2023.50.002.011

    Abstract

    Abstract : Manihot esculenta Crantz, commonly known as cassava, is a staple aliment that is a significant source of revenue for farmers. The embryogenic callus is crucial in the genetic engineering of various crop species, including cassava. Four cultivar cassava landraces from East Java were assessed for their ability to produce friable embryogenic callus (FEC) for protoplast isolation. In this study, four cassava cultivars; (Kaspro, Kuning, Gajah, and Gendruwo); were used to obtain FEC, which involved the culture of immature leaf lobes (ILLs) and apical buds (ABs) media containing MS supplemented with 33 μM picloram and 2 μ M of CuSO4 (M1) or MS supplemented with 50 μM 2,4-D and 2 μM CuSO4 (M2). The highest FEC induction efficiency ranged from 72% to 57%, and the highest FEC number ranged from 4.7 to 3.7 with AB explants in media containing MS + 33 μM pilocram and 2 μM CuSO4 (M1). On the other hand, the efficiency of somatic embryogenesis induction ranged from 67% to 53%, and the number ranged from 4.4 to 3.4. The efficiencies of FEC induction ranged from 48% to 42%, and the number ranged from 3.1 to 2.6 with AB explants in media containing MS + 50 μM 2,4-D and 2 μM CuSO4 (M2); the efficiency of FEC induction ranged from 56% to 50%, and the value ranged from 3.6 to 2.4 with ILL explants. The FEC induction of the Gendruwo cultivar, which was examined using AB and ILL explants, demonstrated the lowest efficiency. Nevertheless, all four cultivars showed the ability to generate FEC, even though their effectiveness differed depending on the explant genotype and the applied media.

  • Research ArticleMarch 29, 2023

    0 607 201

    Embryogenesis and plant regeneration of Panax ginseng Meyer via anther culture and ploidy assessment using flow cytometry

    Jung-Woo Lee ・Kyong-Hwan Bang ・Dong-Hwi Kim・Jang-Uk Kim ・Young-Chang Kim・Ick-Hyun Jo

    J Plant Biotechnol (2023) 50:019-026

    https://doi.org/10.5010/JPB.2023.50.003.019

    Abstract

    Abstract : Korean ginseng (Panax ginseng Meyer) is an economically important plant because of it is rich in saponins. It is mainly cultivated in Asia, including Korea and China. Since ginseng requires a long breeding period due to juvenility, homozygote production techniques, such as anther culture, must be urgently established. In the present study, callus induction and embryogenesis through anther culture were observed in P. ginseng. Murashige and Skoog medium was used as the basal medium suitable for callus induction. When the medium was supplemented with 3% sucrose, the callus induction rate was high and the callus size was large. Cold pretreatment did not significantly affect callus induction and embryogenesis. Embryogenesis was the most efficient when the embryo-formation medium was supplemented with 1.0 or 3.0 mg/L 2,4-dichlorophenoxyacetic acid. Cultivar significantly affected anther culture efficiency. Specifically, ‘Cheongseon’ showed the highest embryo-formation efficiency, whereas no embryogenesis occurred in ‘Sunun’. Ploidy assessment revealed the haploid status of the induced calli. Embryos derived from anther culture formed shoots upon transfer to germination medium, although no difference in ploidy was noted between the induced callus and control. Overall, the anther culture conditions established in the present study may contribute to the production of homozygous P. ginseng plants in the future.

  • Research ArticleMarch 29, 2023

    0 547 176
    Abstract

    Abstract : Zizyphus jujube is a plant in the buckthorn family (Rhamnaceae) that has been the subject of research into antibacterial, antifungal and anti-inflammatory properties of its fruit and seed. However, few studies have investigated its leaves. In this study, the anti-inflammatory activity of ZJL (an extract of Z. jujube leaf) was evaluated to verify its potential as an anti-inflammatory agent and SARS-CoV-2 medicine, using nitric oxide (NO) assay, RT-PCR, SDS-PAGE, Western blotting, and UHPLC/TOFHRMS analysis. We found that ZJL suppresed pro-inflammatory mediators such as NO, inducible nitric oxide synthase (iNOS), cyclooxy-genase-2 (COX-2), and tumor necrosis factor α (TNF-α) in lipopolysaccharide (LPS)-induced RAW264.7 cells. ZJL acted by inhibiting NF-KB and MAPK signaling pathway activity. We also confirmed that ZJL contains a phenol compound and flavonoids with anti-inflammatory activity such as trehalose, maleate, epigallocatechin, hyperoside, catechin, 3-O-coumaroylquinic acid, rhoifolin, gossypin, kaempferol 3-neohesperidoside, rutin, myricitrin, guaiaverin, quercitrin, quercetin, ursolic acid, and pheophorbide a. These findings suggest that ZJL may have great potential for the development of anti-inflammatory drugs and vaccines via inhibition of NF‐ĸB and MAPK signaling in LPS-induced RAW264.7 cells.

  • Research ArticleApril 26, 2023

    0 409 81
    Abstract

    Abstract : The tetraploid Solanum hjertingii, a wild tuber-bearing species from Mexico is a relative of potato, S. tuberosum. The species has been identified as a potential source of resistance to blackening for potato breeding. It does not exhibit enzymatic browning nor blackspot which are physiological disorders. However, due to their sexual incompatibility, somatic hybridization between S. hjertingii and S. tuberosum must be used to introduce various traits from this wild species into potato. After somatic hybridization, molecular markers are essential for selecting fusion products. In this study, the chloroplast genome of S. hjertingii was sequenced by next-generation sequencing technology and compared with those of other Solanum species to develop specific markers for S. hjertingii. The chloroplast genome has a total sequence length of 155,545 bp, and its size, gene content, order and orientation are similar to those of the other Solanum species. Phylogenic analysis including 15 other Solanaceae species grouped S. hjertingii with S. demissum, S. hougasii, and S. stoloniferum. After detailed comparisons of the chloroplast genome sequence with eight other Solanum species, we identified one InDel and seven SNPs specific to S. hjertingii. Based on these, five PCR-based markers were developed for discriminating S. hjertingii from other Solanum species. The results obtained in this study will aid in exploring the evolutionary aspects of Solanum species and accelerating breeding using S. hjertingii.

  • Research ArticleApril 26, 2023

    0 777 128
    Abstract

    Abstract : The diploid Solanum cardiophyllum, a wild tuberbearing species from Mexico is one of the relatives to potato, S. tuberosum. It has been identified as a source of resistance to crucial pathogens and insects such as Phytophthora infestans, Potato virus Y, Colorado potato beetle, etc. and is widely used for potato breeding. However, the sexual hybridization between S. cardiophyllum and S. tuberosum is limited due to their incompatibility. Therefore, somatic hybridization can introduce beneficial traits from this wild species into the potato. After somatic hybridization, selecting fusion products using molecular markers is essential. In the current study, the chloroplast genome of S. cardiophyllum was sequenced by next-generation sequencing technology and compared with those of other Solanum species to develop S. cardiophyllum-specific markers. The total length of the S. cardiophyllum chloroplast genome was 155,570 bp and its size, gene content, order and orientation were similar to those of the other Solanum species. Phylogenic analysis with 32 other Solanaceae species revealed that S. cardiophyllum was expectedly grouped with other Solanum species and most closely located with S. bulbocastanum. Through detailed comparisons of the chloroplast genome sequences of eight Solanum species, we identified 13 SNPs specific to S. cardiophyllum. Further, four SNP-specific PCR markers were developed for discriminating S. cardiophyllum from other Solanum species. The results obtained in this study would help to explore the evolutionary aspects of Solanum species and accelerate breeding using S. cardiophyllum.

  • Research ArticleApril 28, 2023

    0 736 118
    Abstract

    Abstract : Transgenic lilies have been obtained using Agrobacterium tumefaciens (AGL1) with the plant scale explants, followed by DL-phosphinothricin (PPT) selection. In this study, scales of lily plants cv. “red flame” were transformed with the pCAMBIA3301 vector containing the gus gene as a reporter and the blpR gene as a selectable marker, as well as a gene of interest showing herbicide tolerance, both driven by the CaMV 35S promoter. Using a 20-minute infection time and a 5-day cultivation period, factors that optimized and demonstrated a high transformation efficiency were achieved. With these conditions, approximately 22-27% efficiency was observed for Agrobacterium-mediated transformation in lilies. After transformation with Agrobacterium, scales of lilies were transferred to MS medium without selective agents for 2 weeks. They were then placed on selection MS medium containing 5 mg/L PPT for a month of further selection and then cultured for another 4-8 weeks with a 4-week subculture regime on the same selection medium. PPT-resistant scales with shoots were successfully rooted and regenerated into plantlets after transferring into hormone-free MS medium. Also, most survived putatively transformed plantlets indicated the presence of the blpR gene by PCR analysis and showed a blue color indicating expression of the gus gene. In conclusion, when 100 scales of lily cv. “red flame” are transformed with Agrobacterium, approximately 22-27 transgenic plantlets can be produced following an optimized protocol. Therefore, this protocol can contribute to the lily breeding program in the future.

  • Research ArticleApril 28, 2023

    0 1047 149
    Abstract

    Abstract : This study includes the transformation of genes such as ORE7, the increase of gene expression, and the use of the bar gene as a selectable marker that shows herbicide resistance with Agrobacterium tumefaciens using hypocotyls from the oilseed rape “Youngsan” cultivar. To establish an Agrobacterium transformation system for the production of oilseed rape with a high-yield trait, infection time and co-cultivation period with Agrobacterium were tested. Therefore, when hypocotyls from the oilseed rape “Youngsan” cultivar were infected with Agrobacterium for 20 min and co-cultivated for 3 days, approximately 32-36 putatively transformed hypocotyls with shoots including roots survived from 100 inoculated hypocotyls after 4 weeks of transformation on a selection medium containing 20 mg/L of phosphinothricin (PPT) as a selectable agent. Additionally, a PCR assay was performed to confirm the insertion of target genes and showed the presence of the ORE7 gene as a high-yielding trait and the bar gene as a selectable marker. Treatment with 0.5% (v/v) Basta solution as a selectable agent for 6 days with leaves from transformed oilseed rape expressed the bar gene. Therefore, this study can contribute to the development of special oilseed rapes containing agriculturally useful traits such as herbicide resistance, drought tolerance, high yielding traits, and high oleic acid content.

  • Research ArticleMay 2, 2023

    0 777 129

    Analysis of soyasaponin content and biosynthesis-related gene expression in young pea (Pisum sativum L.) sprouts

    Gang Deok Han ・HanGyeol Lee ・Jae-Hyeok Park ・Young Jae Yun ・Gee Woo Kim・Sangyun Jeong ・ So-Yeon Moon ・Hye-Young Seo ・Young-Cheon Kim・Woo Duck Seo ・Jeong Hwan Lee

    J Plant Biotechnol (2023) 50:070-075

    https://doi.org/10.5010/JPB.2023.50.009.070

    Abstract

    Abstract : In legumes, soyasaponins, one of triterpenoid saponins, are major components of secondary metabolites with a more diverse array of bioactive chemicals. Although the biosynthetic pathway of soyasaponins has been largely studied in soybean, the study on the soyasaponin contents and biosynthesis-related gene expression in pea (Pisum sativum L.) is poorly understood. Here, we found the accumulation of only soyasaponin Bb component in the sprouts of two Korean domestic pea cultivars (Dachung and Sachul). This pattern was consistent with our observation that increased expression of PsUGT73P2 and PsUGT91H4 genes, but not PsCYP72A69, could be responsible for biosynthesis of only soyasaponin Bb in pea by examining their gene expression. However, gradual accumulation of soyasaponin Bb at developmental stages was not consistent with the expression of PsUGT73P2 and PsUGT91H4, suggesting that the changes of their protein activities may affect the accumulation patterns of soyasaponin Bb. We also revealed that the increased expression levels of PsUGT73P2 and PsUGT91H4 during light to dark transition led to increase of soyasaponin Bb contents. Collectively, our results provided a molecular basis of metabolic engineering for enhancing useful soyasaponin Bb metabolites in Korean domestic pea cultivars.

  • Research ArticleMay 12, 2023

    0 540 87

    Differential responses of peroxidases in sweetpotato suspensioncultured cells to cadmium treatment

    Ju Hwan Kim ・Ki Jung Nam ・Kang-Lok Lee ・Yun-Hee Kim

    J Plant Biotechnol (2023) 50:076-081

    https://doi.org/10.5010/JPB.2023.50.010.076

    Abstract

    Abstract : As cultured plant cells can grow in high oxidative stress conditions, they form an excellent system to study antioxidant mechanisms and the mass production of antioxidants. Oxidative stress is a major cause of damage in plants exposed to various types of environmental stress, including heavy metals, such as cadmium (Cd). Heavy metal accumulation can interfere with many cell functions and plant growth. To evaluate the contribution of oxidative stress to Cd-induced toxicity, cultured sweetpotato (Ipomoea batatas) cells were treated with increasing concentrations of Cd (0, 10, 25, and 50 μM) and cultured further. Cell growth was significantly inhibited by 25 and 50 μM of Cd, and the total protein content increased with 50 μM of Cd. Additionally, the activity of peroxidase (POD) and ascorbate peroxidase (APX), antioxidant enzymes that remove hydrogen peroxide (a reactive oxygen species), increased in the cells after treatment with 50 μM of Cd. The expression analysis of POD, APX, and peroxiredoxin (PRX) isolated from sweetpotato cultured cells in a previous study revealed the differential expression of POD in response to Cd. In this study, the expression levels of several acidic POD (swpa2, swpa3, and swpa4) and basal POD (swpb1, swpb2, and swpb3) genes were increased in Cd-treated cultured cells. These results indicate that Cd-mediated oxidative stress is closely linked to improved POD-mediated antioxidant defense capacity in sweetpotato suspension-cultured cells.

JPB
Vol 51. 2024

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