Plants are often exposed to biotic and abiotic stresses that affect plant growth, development, and productivity. Drought is an important abiotic stress that has a particularly serious impact on plant growth and development. We transformed rice with PsGPD using Agrobacterium-mediated transformation. We generated independent PsGPD-homozygous transgenic rice plants selected as single copy/intergenic lines by the TaqMan copy number assay and by T-DNA flanking sequences. These transgenic rice plants showed improvement of drought tolerance compared to wild-type plants under drought condition. RNA sequencing analysis showed that 2,992 genes were transcriptionally affected by the PsGPD transgene or drought treatment. In total, 145 genes were modulated by the PsGPD transgene before and after drought treatment. Among these candidate genes, 4 were up- and downregulated in all four comparisons. Several genes, including Os04t0576900, Os03t0629800, and Os04t0518400 (OsPAL7), were involved in tetrapyrrole synthesis. Os09t0522200 (DREB1A), an important component in hormone signal transduction, is a transcription factor (TF) gene that plays vital roles in stress responses. We partially characterized the functions of PsGPD in the drought stress response and the role of major TFs in the drought tolerance mechanism. These genes will be useful targets for both future research and the breeding of drought tolerance in rice.

AT-hook proteins are known to co-regulate transcription of genes through the modification of chromatin architecture. In plants, many genes encoding AT-hook proteins have been shown to be associated with increased seed yield or delayed senescence. In this study, we produced transgenic soybean plants overexpressing chromatin architecture-controlling ATPG8 and ATPG10 genes by Agrobacterium-mediated transformation and examined their agronomic traits to identify the yield increase in soybean crop similar to those seen in model plants, Arabidopsis. A total of 16 (3 of pB2GW7.0-ATPG8 and 13 of pCSEN-ATPG10 transformed) transgenic soybean plants were produced and their T₁ seeds were harvested. Healthy and well-grown transgenic lines were selected (lines #1 and #2 from pB2GW7.0-ATPG8, and lines #8 and #9 from pCSEN-ATPG10), and the insertion and transcription level of genes were confirmed by PCR and RT-PCR with expected size. Investigation on agricultural traits confirms the increase in yield, plant height, the number of pods, and total seed weight with statistical significance when compared to wild-type soybean plants. The yield component study suggested that overexpression of ATPG8 and ATPG10 genes conferred positive effect on yield in transgenic soybean.

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  Phetole Mangena
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  Ji Hyeon Song, Gilok Shin, Hye Jeong Kim, Saet Buyl Lee, Ju Yeon Moon, Jae Cheol Jeong, Hong-Kyu Choi, In Ah Kim, Hyeon Jin Song, Cha Young Kim, Young-Soo Chung
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• Increased Production of α-Linolenic Acid in Soybean Seeds by Overexpression of Lesquerella FAD3-1
  Wan Woo Yeom, Hye Jeong Kim, Kyeong-Ryeol Lee, Hyun Suk Cho, Jin-Young Kim, Ho Won Jung, Seon-Woo Oh, Sang Eun Jun, Hyun Uk Kim, Young-Soo Chung
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AT-hook proteins of plant have shown to be involved in growth and development through the modification of chromatin architecture to co-regulate transcription of genes. Recently, many genes encoding AT-hook protein have been identified and their involvement in senescence delay is investigated. In this study, soybean transgenic plants overexpressing chromatin architecture-controlling ATPG7 gene was produced by Agrobacterium-mediated transformation and investigated for the positive effect on the important agronomic traits mainly focusing on yield-related components. A total of 27 transgenic soybean plants were produced from about 400 explants. T₁ seeds were harvested from all transgenic plants. In the analysis of genomic DNAs from soybean transformants, ATPG7 and Bar fragments were amplified as expected, 975 bp and 408 bp in size, respectively. And also exact gene expression was confirmed by reverse transcriptase-PCR (RT-PCR) from transgenic line #6, #7 and #8. In a field evaluation of yield components of ATPG7 transgenic plants (T₃), higher plant height, more of pod number and greater average total seed weight were observed with statistical significance. The results of this study indicate that the introduction of ATPG7 gene in soybean may have the positive effect on yield components.

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• Efficient Isolation and Gene Transfer of Protoplast in Korean Soybean (Glycine Max (L.) Merr.) Cultivars
  Chuloh Cho, Dool-Yi Kim, Man-Soo Choi, Mina Jin, Mi-Suk Seo
  Korean Journal of Breeding Science.2021; 53(3): 230.     CrossRef
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  Wan Woo Yeom, Hye Jeong Kim, Kyeong-Ryeol Lee, Hyun Suk Cho, Jin-Young Kim, Ho Won Jung, Seon-Woo Oh, Sang Eun Jun, Hyun Uk Kim, Young-Soo Chung
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Plants have adapted to environmental challenges by expressing many plant genes in response to the stresses. Among those genes, CCCH zinc finger proteins are involved in abiotic and biotic stresses. Transgenic soybean plants overexpressing AtSZF2 were produced to investigate that its ectopic overexpression enhanced salt stress tolerance by Agrobacterium-mediated transformation using half-seed explants. Sixteen transgenic lines were chosen to analyze for T-DNA insertion and transcription levels, and most of them were confirmed as positive. In further analysis with Southern blot, stable transformation event and copy number were confirmed. Following high salinity stress on the detached leaf and whole plant of two transgenic lines (#4 and #6) revealed that the ectopic expression of AtSZF2 was correlated with stress tolerance in phenotype, ion leakage and chlorophyll content with statistical significance. In another test with 20% PEG treatment, similar tolerance of transgenic plants was observed with lower ion leakage and higher chlorophyll content, indicating that the damage of cell membrane was prevented in transgenic plants. Finally, expression of various abiotic stress-responding genes was detected by reverse transcriptase and quantitative real-time PCR analysis with the transgenic plants. It could be proposed that introduction of AtSZF2 resulted in the modulation of ABA/stress responsive gene expression in transgenic soybean plants and make them tolerant against salt stress. Considering soybean as a salt-sensitive crop and importance of salt stress tolerance in specific farming region, the introduction of AtSZF2 may provide an approach for crop improvement in soybean breeding.

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An Agrobacterium-mediated transformation system was developed for the ornamental cherry species Prunus incisa. This system uses both an antibiotic resistance gene (NPTII) and a visible selectable marker, the green fluorescent protein (GFP), to select plants. Cells from leaf and root explants were transformed with a NPTII/GFP fusion gene, and selected visually using fluorescence microscopy. Transformed cells were then induced to undergo embryogenesis and reselected by growing on media containing kanamycin. The presence of the GFP/NPTII fusion gene in all parts of transgenic plants grown in the greenhouse for one year was confirmed by PCR and Southern blot analysis. This transformation and selection system will be useful in future work to introduce genes for pathogen resistance and ornamental traits into flowering cherry germplasm.

This study aimed to establish a positive selection system for the genetic transformation of cauliflower using the phosphomannose isomerase (PMI) gene. PMI catalyzes the reversible interconversion of mannose 6-phosphate and fructose 6-phosphate. Thus, only plant cells transformed with the PMI gene can survive on synthetic medium containing mannose as a carbon source. Cotyledon explants from 6-day-old seedlings were infected with Agrobacterium tumefaciens strain LBA4404 harboring binary vector pNWB-JMT containing the jasmonic acid carboxyl methyltransferase (JMT) gene with full codon modification. After co-cultivation with Agrobacterium, positive selection was conducted on MS medium supplemented with 2 mg/L of BAP, 1 mg/L of NAA, 0.6% (w/v) mannose, and 2% (w/v) sucrose. Over 40 putative transgenic plants were obtained in the repetitive transformation experiments. The overall transformation efficiency was 1.2% in the mannose-based selection. Polymerase chain reaction along with Southern and Northern blotting analyses were used to confirm and characterize the transgenic plants containing the integrated JMT gene. After soil acclimatization, transgenic plants were successfully grown to maturity in a greenhouse. T₁ seeds were successfully collected from these transgenic plants. This is the first successful report of cabbage transformation using a mannose-based selection system. Therefore, the positive selection system established in this study could be applied as an alternative tool for the rapid selection of elite lines for the purposes of breeding and reduction of antibiotic use in order to improve human health and environmental safety.

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• Development of an efficient transformation system in eggplant (Solanum melongena L.) using the phosphomannose isomerase (PMI) gene as the selectable marker
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Rice (Oryza sativa L.), as a cereal grain, is the most widely consumed staple food for a large part of the world’s human population especially in Asia. Three indica rice (Oryza sativa L.) cultivars namely BRRI dhan29, BRRI dhan56 and BRRI dhan57 were used in this study to investigate the regeneration ability in rice. Regeneration potential was found to be highest (80.00%) for BRRI dhan29 and lowest (70.00%) for BRRI dhan57. Consequently, the highly regenerating indica rice cultivar BRRI dhan29 was used for genetic transformation. Embryogenic calli induced after 20 days were used for genetic transformation in the experiment. Agrobacterium strain LBA4404 was transformed with pBI121 binary vector which contains kanamycin resistance gene as a selectable marker gene and GUS as a reporter gene. Calli infected with this strain were analyzed by using 5-bromo 4-chloro 3-indolyl- D glucuronide (X-gluc) as a substrate. Expression of gene was determined by using the transformed shoots and roots. Stable integration and expression of GUS gene were also confirmed by using PCR analysis. The frequency of transformation in terms of transient GUS assay was found to be 35.0 ± 2 S.E. This study will provide valuable information for genetic transformation in cereal crops.

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