• Journal of Life Science
• /
• v.26 no.10
• /
• pp.1121-1129
• /
• 2016

High-molecular-weight glutenin subunits (HMW-GSs) are extremely important determinants of the functional properties of wheat dough. Transgenic rice plants containing a wheat TaGlu-Ax1 gene encoding a HMG-GS were produced from the Korean wheat cultivar ‘Jokyeong’ and used to enhance the bread-making quality of rice dough using the Agrobacterium-mediated co-transformation method. Two expression cassettes with separate DNA fragments containing only TaGlu-Ax1 and hygromycin phosphotransferase II (HPTII) resistance genes were introduced separately into the Agrobacterium tumefaciens EHA105 strain for co-infection. Rice calli were infected with each EHA105 strain harboring TaGlu-Ax1 or HPTII at a 3:1 ratio of TaGlu-Ax1 and HPTII. Among 210 hygromycin-resistant T₀ plants, 20 transgenic lines harboring both the TaGlu-Ax1 and HPTII genes in the rice genome were obtained. The integration of the TaGlu-Ax1 gene into the rice genome was reconfirmed by Southern blot analysis. The transcripts and proteins of the wheat TaGlu-Ax1 were stably expressed in rice T₁ seeds. Finally, the marker-free plants harboring only the TaGlu-Ax1 gene were successfully screened in the T₁ generation. There were no morphological differences between the wild-type and marker-free transgenic plants. The quality of only one HMW-GS (TaGlu-Ax1) was unsuitable for bread making using transgenic rice dough. Greater numbers and combinations of HMW and LMW-GSs and gliadins of wheat are required to further improve the processing qualities of rice dough. TaGlu-Ax1 marker-free transgenic plants could provide good materials to make transgenic rice with improved bread-making qualities.

• Plant Biotechnology Reports
• /
• v.4 no.3
• /
• pp.213-222
• /
• 2010

Within their natural habitat, crops are often subjected to drought and heat stress, which suppress crop growth and decrease crop production. Causing overaccumulation of glycinebetaine (GB) has been used to enhance the crop yield under stress. Here, we investigated the response of wheat (Triticum aestivum L.) photosynthesis to drought, heat stress and their combination with a transgenic wheat line (T6) overaccumulating GB and its wild-type (WT) Shi4185. Drought stress (DS) was imposed by controlling irrigation until the relative water content (RWC) of the flag leaves decreased to between 78 and 82%. Heat stress (HS) was applied by exposing wheat plants to $40^{\circ}C$ for 4 h. A combination of drought and heat stress was applied by subjecting the drought-stressed plants to a heat stress as above. The results indicated that all stresses decreased photosynthesis, but the combination of drought and heat stress exacerbated the negative effects on photosynthesis more than exposure to drought or heat stress alone. Drought stress decreased the transpiration rate (Tr), stomatal conductance (Gs) and intercellular $CO_2$ concentration (Ci), while heat stress increased all of these; the deprivation of water was greater under drought stress than heat stress, but heat stress decreased the antioxidant enzyme activity to a greater extent. Overaccumulated GB could alleviate the decrease of photosynthesis caused by all stresses tested. These suggest that GB induces an increase of osmotic adjustments for drought tolerance, while its improvement of the antioxidative defense system including antioxidative enzymes and antioxidants may be more important for heat tolerance.

• Journal of Practical Agriculture & Fisheries Research
• /
• v.25 no.4
• /
• pp.138-147
• /
• 2024

Agrobacterium-mediated transformation (AMT) is a method that allows for the stable integration of DNA fragments into the plant genome. Transgenic plants generated through AMT typically exhibit a lower copy number of the transgene compared to those induced by particle bombardment. Furthermore, AMT offers a straightforward and efficient approach for generating transgenic plants. While the transformation efficiency of wheat is comparatively lower than that of other monocot plants such as Rice (Oryza sativa L.) and Maize (Zea mays L.), the cultivars 'Bobwhites' and 'Fielder' are commonly employed for wheat transformation. To date, there have been no reported instances of successful development of transgenic plants using Korean wheat varieties through AMT. This study aims to assess the transformation efficiency of 43 Korean wheat cultivars using the GUS assay, with the goal of identifying suitable Korean wheat cultivars for AMT. The pCAMBIA1301 vector, carrying the β-glucuronidase (GUS) gene, was incorporated into Agrobacterium strain EH105. Following the inoculation of Agrobacterium into immature embryos, GUS assays were conducted 'Saeol', 'Jopum', and 'Jonong' showed 100% (the number of embryos showing GUS spots/the number of embryos used for AMT) among 43 cultivars. In addition, cultivars with more than 70% were 'Saekeumgang', 'Jojung', 'Tapdong', 'Anbaek', 'Dabun', 'Sugang', 'Keumgang', 'Jeokjung', 'Seodun', 'Joeun', 'Dajung', and 'Baekjung'. It seems that the 15 cultivars above showed the possibility of using AMT. On the other hand, 'Yeonbaek', 'Goso', 'Baekgang', and 'Johan' showed less than 20% and GUS spots were not observed in 'Gru', 'Gobun', 'Milseong', and 'Shinmichal-1'. This study explores transient GUS expression in Korean wheat cultivars seven days after AMT. The observed initial high efficiency of transient transformation suggests the potential for subsequent stable transformation efficiency. Korean wheat cultivars demonstrating elevated transient transformation efficiency could serve as promising candidates for the development of stable transgenic wheat.

• Journal of Life Science
• /
• v.24 no.6
• /
• pp.618-625
• /
• 2014

Rice flour is used in many food products. However, dough made from rice lacks extensibility and elasticity, making it less suitable than wheat for many food products such as bread and noodles. The high-molecular weight glutenin subunits (HMW-GS) of wheat play a crucial role in determining the processing properties of the wheat grain. This paper describes the development of marker-free transgenic rice plants expressing a wheat Glu-Dy10 gene encoding the HMG-GS from the Korean wheat cultivar 'Jokyeong' using Agrobacterium-mediated co-transformation. Two expression cassettes, consisting of separate DNA fragments containing Glu-1Dy10 and hygromycin phosphotransferase II (HPTII) resistance genes, were introduced separately into Agrobacterium tumefaciens EHA105 for co-infection. Each EHA105 strain harboring Glu-1Dy10 or HPTII was infected into rice calli at a 3: 1 ratio of Glu-1Bx7 and HPTII. Among 290 hygromycin-resistant $T_0$ plants, we obtained 29 transgenic lines with both the Glu-1Dy10 and HPTII genes inserted into the rice genome. We reconfirmed the integration of the Glu-1Dy10 gene into the rice genome by Southern blot analysis. Transcripts and proteins of the Glu-1Dy10 in transgenic rice seeds were examined by semi-quantitative RT-PCR and Western blot analysis. The marker-free plants containing only the Glu-1Dy10 gene were successfully screened in the $T_1$ generation.

• Journal of Life Science
• /
• v.23 no.11
• /
• pp.1317-1324
• /
• 2013

High-molecular weight glutenin subunits (HMW-GS) have been shown to play a crucial role in determining the processing properties of the wheat grain. We have produced marker-free transgenic rice plants containing a wheat Glu-1Bx7 gene encoding the HMG-GS from the Korean wheat cultivar 'Jokyeong' using the Agrobacterium-mediated co-transformation method. The Glu-1Bx7-own promoter was inserted into a binary vector for seed-specific expression of the Glu-1Bx7 gene. Two expression cassettes comprised of separate DNA fragments containing only Glu-1Bx7 and hygromycin phosphotransferase II (HPTII) resistance genes were introduced separately to the Agrobacterium tumefaciens EHA105 strain for co-infection. Each EHA105 strain harboring Glu-1Bx7 or HPTII was infected to rice calli at a 3:1 ratio of Glu-1Bx7 and HPTII, respectively. Then, among 216 hygromycin-resistant $T_0$ plants, we obtained 24 transgenic lines with both Glu-1Bx7 and HPTII genes inserted into the rice genome. We reconfirmed integration of the Glu-1Bx7 gene into the rice genome by Southern blot analysis. Transcripts and proteins of the wheat Glu-1Bx7 were stably expressed in the rice $T_1$ seeds. Finally, the marker-free plants harboring only the Glu-1Bx7 gene were successfully screened at the $T_1$ generation.

• Asian-Australasian Journal of Animal Sciences
• /
• v.16 no.12
• /
• pp.1732-1737
• /
• 2003

An experiment was conducted to investigate the effect of feeding transgenic cottonseed (Bt.) vis-a-vis non-transgenic (non-Bt.) cottonseed on blood biochemical constituents in lactating Murrah buffaloes. Twenty Murrah buffaloes in mid-lactation were divided into 2 groups of 10 each. Animals of group I were fed with 39.5% non-transgenic cottonseed in concentrate mixture while the same percentage of transgenic (Bt.) cottonseed was included in the concentrate mixture fed to the animals of group II. Animals of both groups were fed with concentrate mixture to support their milk production requirements. Each buffalo was also offered 20 kg mixed green fodder (oats and berseem) and wheat straw ad libitum. The experimental feeding trial lasted for 35 days. There was no significant difference in the dry matter intake between the two groups of buffaloes. All the buffaloes gained body weight, however, the differences were non significant. Total erythrocyte count, hemoglobin content and packed cell volume were $9.27{\pm}0.70${\times}10^6/{\mu}l$, $13.01{\pm}0.60gdl$ and $34.87{\pm}1.47%$, respectively in group I with the corresponding figures of $8.88{\pm}0.33$, $12.99{\pm}0.52$ and $31.08{\pm}1.52$ in group II. The values of total erythrocyte count, haemoglobin content and packed cell volume did not differ significantly between the two groups of buffaloes. The concentration of plasma glucose, serum total proteins, albumin, globulin, triglycerides and high density lipoprotein were non significantly higher in buffaloes fed non-transgenic cottonseed than in buffaloes fed transgenic cottonseed. The cholesterol concentration was significantly (p<0.01) higher in buffaloes of group I ($136.84{\pm}8.40mg/dl$) than in buffaloes of group II ($105.20{\pm}1.85mg/dl$). The serum alkaline phosphotase, glutamic-oxaloacetate transaminase and glutamic-pyruate transaminase activities did not differ significantly between two groups of buffaloes. However, serum glutamic-pyruate transaminase activity was considerably high in buffaloes fed nontransgenic cottonseed as compared to buffaloes fed transgenic cottonseed. Bt. proteins in serum samples of animals of group II were not detected after 35 days of feeding trial. It was concluded that transgenic cottonseed and non-transgenic cottonseed have similar nutritional value without any adverse effects on health status of buffaloes as assessed from haematobiochemical constituents.

• Journal of Plant Biotechnology
• /
• v.48 no.4
• /
• pp.246-254
• /
• 2021

Environmental stresses are estimated to have reduced global crop yields of wheat by 5.5%. However, traditional approaches for the transfer of resistance to these stresses in wheat plants have yielded limited results. In this regard, genetic transformation has undoubtedly opened up new avenues to overcome crop losses due to various abiotic stresses. Particle bombardment has been successfully employed for obtaining transgenic wheat. However, most of these procedures employ immature embryos, which are not available throughout the year. Therefore, the present investigation utilized mature seeds as the starting material and used the calli raised from three Moroccan durum wheat varieties as the target tissue for genetic transformation by the biolistic approach. The pANIC-5E plasmid containing the SINA gene for drought and salinity tolerance was used for genetic transformation. To enhance the regeneration capacity and transformation efficiency of the tested genotypes, the study compared the effect of copper supplementation in the induction medium (up to 5 μM) with the standard MS medium. The results show that the genotypes displayed different sensitivities to CuSO₄, indicating that the transformation efficiency was highly genotype-dependent. The integration of transgenes in the T₀ transformants was demonstrated by polymerase chain reaction (PCR) analysis of the obtained resistant plantlets with primers specific to the SINA gene. Among the three genotypes studied, 'Isly' showed the highest efficiency of 9.75%, followed by 'Amria' with 1.25% and 'Chaoui' with 1%.

• Proceedings of the Korean Society of Crop Science Conference
• /
• 2022.10a
• /
• pp.189-189
• /
• 2022

Soil salinity negatively affects plant growth, productivity, and metabolism. Rice is known to have more sensitive phenotypes than other cereal crops, such as wheat, sorghum, and barley. We characterized the molecular function of rice C3HC4 as a really interesting new gene (RING). Oryza sativa RING finger protein H2-3 (OsRFPH2-3) was highly expressed in 100 mM NaCl. To identify the localization of OsRFPH2-3, we fused vectors that include C-terminal GFP protein (35S;;OsRFPH2-3-GFP). OsRFPH2-3 was expressed in the nucleus in rice protoplasts. An in vitro ubiquitin assay demonstrated that OsRFPH2-3 possessed E3-ubiquitin ligase activity. However, the mutated OsRFPH2-3 were not possessed any E3-ubiquitin ligase activity. Under normal conditions, there is no significant phenotypic difference between transgenic plants and WT plants. However, OsRFPH2-3-overexpressing plants exhibited higher fresh weight and length under saline conditions. Also, transgenic plants maintain higher chlorophyll, proline, and soluble sugar contents and lower H₂O₂ and MDA contents than the wild type; these results support transgenic plants with enhanced salinity tolerance phenotypes.

• Journal of Plant Biotechnology
• /
• v.3 no.2
• /
• pp.67-81
• /
• 2001

Traditionally, genetic variability is generated by an extensive crossing program, which is complemented by strict selection to identify useful new recombinants. Plant biotechnology offers many opportunities for breeders to solve certain breeding problems at the molecular level. The tissue culture methodology and the genetic modification of economically important monocotyledons have undergone a revolution in the last decade. As the production of transgenic plants is a complex procedure, including the uptake of DNA molecules into the cells, the integration of foreign nucleotide sequences into the host genomic DNA and the expression of new genes in a controlled way, and as there are still many unsolved questions, further development is necessary. The methodology opens up the possibility of introducing novel genes that may induce resistance to diseases and abiotic stresses, allow the modification of dough quality and the dietetic quality of proteins, and increase the levels of micronutrients such as iron, zinc, and vitamins. In the present review, the authors would like to summarise the most important advances in wheat transformation.

• Journal of Life Science
• /
• v.22 no.9
• /
• pp.1152-1158
• /
• 2012

Development of transgenic plant increasing crop yield or disease resistance is good way to solve the world food shortage. However, the persistence of marker genes in crops leads to serious public concerns about the safety of transgenic crops. In the present paper, we developed marker-free transgenic rice inserted high molecular-weight glutenin subunit (HMW-GS) gene ($D{\times}5$) from the Korean wheat cultivar 'Jokyeong' using Agrobacterium-mediated co-transformation method. Two expression cassettes comprised of separate DNA fragments containing only the $D{\times}5$ and hygromycin resistance (HPTII) genes were introduced separately into Agrobacterium tumefaciens EHA105 strain for co-infection. Each EHA105 strain harboring $D{\times}5$ or HPTII was infected into rice calli at a 3: 1 ratio of EHA105 with $D{\times}5$ gene and EHA105 with HPTII gene expressing cassette. Then, among 66 hygromycin-resistant transformants, we obtained two transgenic lines inserted with both the $D{\times}5$ and HPTII genes into the rice genome. We reconfirmed integration of the $D{\times}5$ and HPTII genes into the rice genome by Southern blot analysis. Wheat $D{\times}5$ transcripts in $T_1$ rice seeds were examined with semi-quantitative RT-PCR. Finally, the marker-free plants containing only the $D{\times}5$ gene were successfully screened at the $T_1$ generation. These results show that a co-infection system with two expression cassettes could be an efficient strategy to generate marker-free transgenic rice plants.