Soo-Hyun Lim

Phenotypic Characterization of Transgenic Miscanthus sinensis Plants Overexpressing Arabidopsis Phytochrome B

Phytochromes are dimeric pigment proteins with reversible photochromism between red and far-red light-absorbing forms. They are photoreceptors that regulate various aspects of plant growth and development and have been used for biotechnological applications to improve agricultural performance of crops. Miscanthus species have been suggested as one of the most promising energy crops. In this paper, Arabidopsis phytochrome B (PHYB) gene was introduced into Miscanthus sinensis using Agrobacterium-mediated transformation method that we developed recently, with the herbicide resistance gene (BAR) as a selection marker. After putative transgenic plants were selected using the herbicide resistance assay, genomic integration of the transgene was confirmed by genomic PCR and Southern blot analysis, and transgene expression was validated by Northern blot analysis. Compared to nontransformed control plants, transgenic plants overexpressing PHYB showed phenotypes with increased phytochrome B function, which includes increased chlorophyll content, decreased plant height, and delayed flowering. Therefore, these results suggest that Arabidopsis phytochrome B is functional in M. sinensis and provide a method to develop Miscanthus varieties with enhanced agricultural performance using phytochromes.

Prediction of Seedling Emergence and Early Growth of Eleocharis kuroguwai Ohwi under Evaluated Temperature

Field and pot experiments were conducted to investigate seedling emergence and early growth of Eleocharis kuroguwai panted on different dates. Non-linear regression analyses of observed data against effective accumulated temperature (EAT) with the Gompertz model showed that the Gompertz model works well in describing seedling emergence and early growth of E. kuroguwai regardless of planting date and soil burial depth. EATs required for 50% of the maximum seedling emergence of E. kuroguwai planted at 1, 3 and 5 cm soil burial depth in the pot experiment were estimated to be 54.5, 84.0 and 118.0℃, respectively, and 56.7℃ when planted at 1 cm in the field experiment. EATs required for 50% of the maximum leaf number of E. kuroguwai planted at 1, 3 and 5 cm soil burial depth in the pot experiment were estimated to be 213.3, 249.0 and 291.6℃, respectively, and 239.5℃ when planted at 1 cm in the field experiment. Therefore, models developed in this study thus predicted that if rotary tillage with water is made on 27 May under +2℃ elevated temperature condition, dates for 50% of the maximum seedling emergence, 5 leaf stage and 5 cm plant height of E. kuroguwai buried at 3 cm soil depth were predicted to be 2 June, 10 June and 12 June. These dates are 1 day earlier for the seedling emergence and 3 days earlier for the early growth as compared with current temperature condition, suggesting that earlier application of herbicides is required for effective control of E. kuroguwai.

Complete chloroplast genomes of two Miscanthus species

Abstract The complete chloroplast (cp) genomes of two Miscanthus species, M. sinensis and M. sacchariflorus, were sequenced and investigated for genes, genome size variation, and polymorphisms. There are 170 genes in both cp genomes, consisting of 122 mRNA genes (84 protein-coding genes and 38 hypothetical genes), 40 tRNA genes, and 8 rRNA genes. The cp genome contains two inverted repeat (IR) regions, separated by large single copy (LSC) region and small single copy (SSC) region. Indels were responsible for 40 bp difference in cp genome size in two species. In addition, we established phylogenetic relationship with other monocot cp genomes, and estimated divergence time. The two Miscanthus species clustered together among other C4 monocot species and the divergence time of two Miscanthus species was approximately 0.5 1–0.84 Mya.

Evaluation of maximum potential gene flow from herbicide resistant Brassica napus to its male sterile relatives under open and wind pollination conditions

Pollen-mediated gene flow (PMGF) from genetically modified (GM) Brassica napus to its wild relatives by wind and insects is a major ecological concern in agricultural ecosystems. This study conducted is to estimate maximum potential gene flow and differentiate between wind- and bee-mediated gene flows from herbicide resistant (HR) B. napus to its closely-related male sterile (MS) relatives, B. napus, B. juncea and Raphanus sativus. Various markers, including pods formation in MS plants, herbicide resistance, and SSR markers, were used to identify the hybrids. Our results revealed the following: 1) maximum potential gene flow (a maximum % of the progeny of pollen recipient confirmed hybrid) to MS B. napus ranged from 32.48 to 0.30% and from 14.69 to 0.26% at 2-128 m from HR B. napus under open and wind pollination conditions, respectively, and to MS B. juncea ranged from 21.95 to 0.24% and from 6.16 to 0.16%, respectively; 2) estimates of honeybee-mediated gene flow decreased with increasing distance from HR B. napus and ranged from 17.78 to 0.03% at 2-128 m for MS B. napus and from 15.33 to 0.08% for MS B. juncea; 3) a small-scale donor plots would strongly favour insect over wind pollination; 4) no gene flow occurred from HR B. napus to MS R. sativus. Our approach and findings are helpful in understanding the relative contribution of wind and bees to gene flow and useful for estimating maximum potential gene flow and managing environmental risks associated with gene flow.

Prediction of Seedling Emergence of Humulus japonicus

This experiment was conducted to investigate seedling emergence of Humulus japonicus in Seoul and Suwon and to predict its seedling emergence in various locations in Korea. Non-linear regression analysis to fit Gompertz model to accumulated seedling emergence of H. japonicus was performed to describe its seedling emergence. Parameter estimates from the non-linear regression and estimated effective accumulated temperature required for its seedling emergence were further used to predict seedling emergence of H. japonicus in 8 major geographical regions of Korea. Seedling emergence of H. japonicus versus effective accumulated temperature was well described by Gompertz model in both Seoul and Suwon. Effective accumulated temperatures required for the first seedling emergence and 50% of the maximum seedling emergence were estimated to be and , respectively. Therefore, the dates for the first seedling emergence of H. japonicus were predicted to be 21 and 30 March in Daegu and Daejeon, while 4 and 6 April in Suwon and Chuncheon, respectively. The dates reaching 50% of maximum seedling emergence were also predicted to be 25 March in Daegu, while 3, 8 and 9 April in Daejeon, Suwon and Chuncheon, respectively.

Prediction of Seedling Emergence and Early Growth of Monochoria vaginalis and Scirpus juncoides under Elevated Temperature

This experiment was conducted to investigate seedling emergence and early growth of Monochoria vaginalis and Scirpus juncoides in the controlled-environment chamber maintained at different temperatures. Non-linear regression analyses of observed data against effective accumulated temperature (EAT) with the Gompertz and logistic models showed that the Gompertz and logistic models worked well in describing seedling emergence and early growth of both weed species, respectively, regardless of temperature. EATs required for 50% of the maximum seedling emergence and the maximum leaf number of M. vaginalis were estimated to be 69.3 and 131℃, respectively, while those of S. juncoides were 94.8 and 137℃, respectively. Models developed in this study thus were used to predict seedling emergence and early growth under elevated temperature condition. If rotary tillage with water is made on 27 May under +3℃ elevated temperature condition, dates for 50% of the maximum seedling emergence and 4 leaf stage were predicted to be 1 June and 15 June for M. vaginalis and 3 June and 14 June for S. juncoides, respectively. As compared with current temperature, these dates are 1-2 days earlier for the seedling emergence and 3 days earlier for the early growth, suggesting that earlier application of herbicides is required for effective control of M. vaginalis and S. juncoides under elevated temperature condition in the future.

Assessment of potential environmental risks of transgene flow in smallholder farming systems in Asia: Brassica napus as a case study in Korea

The cultivation of genetically modified (GM) crops has raised many questions regarding their environmental risks, particularly about their ecological impact on non-target organisms, such as their closely-related relative species. Although evaluations of transgene flow from GM crops to their conventional crops has been conducted under large-scale farming system worldwide, in particular in North America and Australia, few studies have been conducted under smallholder farming systems in Asia with diverse crops in co-existence. A two-year field study was conducted to assess the potential environmental risks of gene flow from glufosinate-ammonium resistant (GR) Brassica napus to its conventional relatives, B. napus, B. juncea, and Raphanus sativus under simulated smallholder field conditions in Korea. Herbicide resistance and simple sequence repeat (SSR) markers were used to identify the hybrids. Hybridization frequency of B. napus × GR B. napus was 2.33% at a 2 m distance, which decreased to 0.007% at 75 m. For B. juncea, it was 0.076% at 2 m and decreased to 0.025% at 16 m. No gene flow was observed to R. sativus. The log-logistic model described hybridization frequency with increasing distance from GR B. napus to B. napus and B. juncea and predicted that the effective isolation distances for 0.01% gene flow from GR B. napus to B. napus and B. juncea were 122.5 and 23.7 m, respectively. Results suggest that long-distance gene flow from GR B. napus to B. napus and B. juncea is unlikely, but gene flow can potentially occur between adjacent fields where the smallholder farming systems exist.