• KOREAN JOURNAL OF CROP SCIENCE
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• v.49 no.5
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• pp.434-439
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• 2004

Medicago truncatula is a model plant for molecular genetic studies of legumes and plant-microbe interactions. To accelerate finding of genes that play roles in the early stages of nodulation and stress responses, a trans-genic plant was developed that contains a promoter­reporter fusion. The promoter of rip], a Rhizobium-induced peroxidase gene, was fused to the coding region of $\beta-glucuronidase (GUS)$ gene and inserted into a modified plant transformation vector, pSLJ525YN, in which the bar gene was preserved from the original plasmid but the neomycin phosphotransferase gene was replaced by a polylinker. Transformation of M. truncatula was carried out by vacuum infiltration of young seedlings with Agrobacterium. Despite low survival rates of infiltrated seedlings, three independent transformants were obtained from repeated experiments. Southern blot analyses revealed that 7 of 8 transgenic plants of the T 1 generation contained the bar gene whereas 6 $T_1$ plants contained the GUS gene. These results indicate that vacuum infiltration is an effective method for transformation of M. truncatula. The progeny seeds of the transgenic plants will be useful for mutagenesis and identification of genes that are placed upstream and may influence the expression of rip] in cellular signaling processes including nodulation.

• Journal of Plant Biotechnology
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• v.33 no.4
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• pp.263-270
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• 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.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.45 no.5
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• pp.310-315
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• 2000

Alfalfa (Medicago sativa L.) plants have been reported to contain water-soluble substances that are autotoxic as well as allelopathic. Laboratory experiment through a petri-dish assay with imbibed seeds was conducted to evaluate both autotoxic and allelopathic effects of alfalfa leaf extracts on the germination and early seedling growth of alfalfa, red clover, crested wheatgrass, and Russian wildrye. Alfalfa seed germination was delayed dependent on extract concentration, with no difference in final germination at 72 hours. Root growth of alfalfa was stimulated up to 14% above control at very low concentrations of both leaf and stem extracts of alfalfa and was significantly reduced at extract concentration of more than 0.5g dry tissue/L (${gL}_{-1}$). Leaf extracts were generally more autotoxic for root growth than were stem extracts. Hypocotyl growth was not affected by all the concentrations of both leaf and stem extracts. Root length of legumes was more sensitive to the autotoxic chemicals from leaf extracts than was germination or shoot length. Hypocotyl growth of two legume plants and plant height of two grasses were not influenced by extracts. Seed germination and root growth of legumes were more inhibited by aqueous extracts of alfalfa leaf than were those of grasses. This result indicates autotoxic effect of alfalfa leaf extracts seems to be greater than allelopathic effect.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.46 no.6
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• pp.449-452
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• 2001

Legume seed isoflavones may have a variety of desirable physiological effect on the human health including both the circulatory and skeletal systems. The present study was performed to determine the isoflavone content of leaf and seed as well as to purify and identify the types of isoflavone from leaf extract of hyacinth bean (Lablab purpureus (L.) Sweet). Reverse phase HPLC revealed six different types of isoflavone such as daidzin, genistin, daidzein, genistein, 6"-o-acetyl genistin and 6"-o-acetyl daidzin in aqueous methanol extract from seeds and leaves of the hyacinth bean. Relatively, leaf isoflavone content of hyacinth bean was greater than seed isoflavone content. Using DiAion HP-20 silica gel and sephadex LH-20 chromatography, pure daidzein was identified in the ether layer, whereas genistin was in the EtOAC fraction.

• Proceedings of the Botanical Society of Korea Conference
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• 1996.07a
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• pp.61-74
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• 1996

Human and monogastric animals can not synthesize 10 out of the 20 amino asids and therefor need to obtain these from their diet. The plant seed is a major source of dietary protein. It is particular important in their study to increase nutritional quality of the seed storage proteins. The low contents of lysine, asparagine and threonenein various cereal seeds and of cystein and methionine. In legume seeds is due to the low proportions of these amino acids in the major storage proteins, we have tried to apply the three strategies; (1) mutagenesis and selection of specific amino acid analogue resistance, (2) cloning and expression study of lysine biosynthesis related gene, (3) transfomation of lysine rich soybean glycinin gene. The 5-methyltryptophan (5MT) resistant cell lines, SAR1, SAR2 and SAR3 were selected from anther derived callus of rice (Oryza sativa L. "Sasanishiki"). Among these selected cell lines, two (SAR1 and SAR3) were able to grow stably at 200 mg/L of 5MT. Analysis of the freed amino acids in callus shows that 5MT resistant cells (SAR3) accumulated free tryptophan at least up to 50 times higher than those that of the higher than of SAS. These results indicated that the 5MT resistant cell lines are useful in studies of amino acid biosynthesis. Tr75, a rice (Oryza sativa L., var. Sasanishiki) mutant resistant to 5MT was segregated from the progenies of its initial mutant line, TR1. The 5MT resistant of TR75 was inherited in the M8 generations as a single dominant nuclear gene. The content of free amino acids in the TR75 homozygous seeds increased approximately 1.5 to 2.0 fold compared to wild-type seeds. Especially, the contents of tryptophan, phenylalanine and aspartic acid were 5.0, 5.3 and 2.7 times higher than those of wild-type seeds, respectively. The content of lysine is significantly low in rice. The lysine is synthesized by a complex pathway that is predominantly regulated by feedback inhibition of several enzymes including asparginase, aspatate kinase, dihydrodipicolinat synthase, etc. For understanding the regulation mechanism of lysine synthesis in rice, we try to clone the lysine biosynthetic metabolism related gene, DHPS and asparaginase, from rice. We have isolated a rice DHPS genomic clone which contains an ORF of 1044 nucleotides (347 amino acids, Mr. 38, 381 daltons), an intron of 587 nucleotides and 5'and 3'-flanking regions by screening of rice genomic DNA library. Deduced amino acid sequence of mature peptide domain of GDHPS clone is highly conserved in monocot and dicot plants whereas that of transit peptide domain is extremely different depending on plant specie. Southern blot analysis indicated that GDHPS is located two copy gene in rice genome. The transcripts of a rice GDHPS were expressed in leaves and roots but not detected in callus tissues. The transcription level of GDHPS is much higher in leaves indicating enormous chloroplast development than roots. Genomic DNA clones for asparaginase genes were screened from the rice genomic library by using plaque hybridization technique. Twelve different genomic clones were isolated from first and second screening, and 8 of 12 clones were analyzed by restriction patterns and identified by Southern Blotting, Restriction enzyme digestion patterns and Southern blot analysis of 8 clones show the different pattern for asparaginase gene. Genomic Southern blot analysis from rice were done. It is estimated that rice has at least 2-3 copy of asparaginase gene. One of 8 positive clones was subcloned into the pBluescript SK(+) vector, and was constructed the physical map. For transformation of lysine rich storage protein into tobacco, soybean glycinin genes are transformed into tobacco. To examine whether glycinin could be stably accumulated in endosperm tissue, the glycinin cDNA was transcriptionally fused to an endosperm-specific promotor of the rice storage protein glutelin gene and then introduced into tobacco genomic via Agrobacterium-mediated transformation. Consequently the glycinin gene was expressed in a seed-and developmentally-specific manner in transgenic tobacco seeds. Glycinin were targeted to vacuole-derived protein bodies in the endosperm tissue and highly accumulated in the matrix region of many transgenic plant (1-4% of total seed proteins). Synthesized glycinin was processed into mature form, and assembled into a hexamer in a similar manner as the glycinin in soybean seed. Modified glycinin, in which 4 contiguous methionine residues were inserted at the variable regions corresponding to the C - teminal regions of the acidic and basic polypeptides, were also found to be accumulated similarly as in the normal glycinin. There was no apparent difference in the expression level, processing and targeting to protein bodies, or accumulation level between normal and modified glycinin. glycinin.

• Korean Journal of Plant Resources
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• v.31 no.3
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• pp.237-253
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• 2018

The adzuki bean (Vigna angularis L.) is a red-grained legume that has a number of essential nutrients and is used in traditional dishes in Asia. Adzuki bean industrial by-products are also a potential low-cost source of some unique bioactive polyphenols. Hence, here, the authors aimed to perform a comparative study of the phytochemical profiles of the leaves and seeds of the adzuki bean and compare their antioxidant, ${\alpha}$-glucosidase inhibition, and tyrosinase inhibition activity. The authors assessed antioxidant activity by DPPH, ABTS, FRAP, PR, TPC, and SOD assays, which showed wide variation, respectively. From the relative antioxidant capacity index results, 10 adzuki bean landraces were selected to compare for phytochemicals and bioactivity using leaf and seed extracts. Antioxidant, ${\alpha}$-glucosidase inhibition, and tyrosinase inhibition activity in the leaf extracts were higher than in the seed extracts, and there were more flavonols and isoflavones in the leaf extracts than in the seed extracts. This study demonstrated that adzuki bean leaf extracts could be a new natural antioxidant or antidiabetic agent and a skin whitener and can also be used in industrial applications.

• Advances in environmental research
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• v.3 no.3
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• pp.207-216
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• 2014

A study was conducted to determine the phytotoxic effect of mercury on seed germination and seedling growth of an important arid legume tree Albizia lebbeck. The seeds germination and seedling growth performance of A. lebbeck responded differently to mercuric chloride treatment (1 mM, 3 mM, 5 mM and 7 mM) as compared to control. Seed germination of A. lebbeck was significantly (p < 0.05) affected by mercury treatment at 1 mM. Root growth of A. lebbeck was not significantly affected by mercury treatment at 1 mM, and 3 mM. Shoot and root length of A. lebbeck were significantly (p < 0.05) affected by 5 mM concentration of mercury treatment. Increase in concentration of mercury treatment at 5 mM and 7 mM significantly (p < 0.05) reduced seedling dry weight of A. lebbeck. The treatment of mercury at 1 mM decreased high percentage of seed germination (22%), seedling length (10%), root length (21.85%) and seedling dry weight (9%). Highest decrease in seed germination (51%), seedling (34%), root length (48%) and seedling dry weight (41%) of A. lebbeck occurred at 7 mM mercury treatment. A. lebbeck showed high percentage of tolerance (78.14%) to mercury at 1 mM. However, 7 mM concentration of mercury produced lowest percentage of tolerance (51.65%) in A. lebbeck. The seed germination potential and seedling vigor index (SVI) clearly decreased with the higher level of mercury. Plantation of A. lebbeck in mercury-polluted area will help in reducing the burden of mercury pollution. A. lebbeck can serve better in coordinating in land management programs in metal contaminated areas. The identification of the toxic concentration of metals and tolerance indices of A. lebbeck would also be helpful for the establishment of air quality standard.

• Asian-Australasian Journal of Animal Sciences
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• v.12 no.8
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• pp.1205-1214
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• 1999

Whole lupinus albus seeds were pressure toasted at temperatures of 100, 118 and $136^{\circ}C$ for 3, 7, 15 and 30 min to study rumen degradation and post-rumen digestion and to determine optimal heating conditions for the Dutch dairy feed industry. In sacco nylon bag and mobile bag techniques were employed for rumen and intestine incubations to determine ruminal degradation characteristics and intestinal digestion of crude protein (CP) in 4 lactation rumen cannulated and 4 lactating intestinal cannulated Dutch dairy cows fed 47% hay and 53% concentrate according to Dutch dairy requirements. Measured rumen degradation characteristics were soluble fraction (S), undegradable fraction (U), potentially degradable fraction (D), lag time (T0) and rate of degradation (Kd) of insoluble but degradable fraction. Percentage bypass feed protein (BCP), ruminal microbial protein synthesized based on available nitrogen (N_MP) and that based on available energy (E_MP), true protein supplied to the small intestine (TPSI), truly absorbed BCP (ABCP), absorbed microbial protein (AVP) in the small intestine, endogenous protein losses in the digestion (ENDP), true digested protein in the small intestine (TAP or DVE in Dutch) and degraded protein balance (PDB or OEB in Dutch) were totally evaluated using the new Dutch DVE/OEB System. Pressure toasting decreased (p<0.001) rumen degradability of CP. It reduced S (p<0.05) and Kd (p=0.06), increased D (p<0.05) and U (p<0.01) but did not alter T0 (p>0.05), thus resulting in dramatically increased BCP (p<0.001) with increasing time and temperature from 73.7 (raw) up to 182.5 g/kg DM ($136^{\circ}C/15min$). Although rumen microbial protein synthesized based on available energy (E_MP) was reduced, true protein (microbial and bypass feed protein) supplied to the small intestine (TPSI) was increased (p<0.001) from 153.1 (raw) to 247.6 g/kg DM ($136^{\circ}C/15min$). Due to digestibility of BCP in the intestine not changing (p>0.05) average 87.8%, the absorbed BCP increased (p<0.001) from 62.3 (raw) to 153.7 g/kg DM ($136^{\circ}C/15min$). Therefore DVE value of true digested protein in the small intestine was significantly increased (p<0.001) from 118.9 (raw) to 197.0 g/kg DM ($136^{\circ}C/15min$) and OEB value of degraded protein balance was significantly reduced (p<0.001) from 147.2 (raw) to 63.1 g/kg DM ($136^{\circ}C/15min$). It was concluded that pressure toasting was effective in shifting degradation of CP of lupinus albus from the rumen to small intestine without changing intestinal digestion. Further studies are required on the degradation and digestion of individual amino acids and on the damaging effects of processing on amino acids, especially the first limiting amino acids.

• Korean Journal of Plant Resources
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• v.33 no.1
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• pp.50-61
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• 2020

Pigeon pea (Cajanus cajan) is an important legume species that produces seeds that are rich in phenolic compounds and dietary nutrients. The aim of the present study was to evaluate the agricultural characters, phenolic and nutritional contents, and antioxidant capacities of seeds from 82 pigeon pea germplasms cultivated in the Republic of Korea. The accessions exhibited compact (24.39%), semi-spreading (74.39%), or spreading (1.22%) growth habits and determinate (89.02%), indeterminate (8.54%), or semi-determinate (2.44%) flowering patterns. Days to 75% maturity ranged from 30 to 72 d, and yield per plant ranged from 6.00 to 148.60 g. Meanwhile, total phenolic, crude protein, crude fiber, and dietary fiber contents ranged from 16.42 ± 0.62 to 29.67 ± 0.43 mg gallic acid equivalent per g of dried extract, from 16.76 ± 6.74% to 22.61 ± 0.05%, from 4.70 ± 0.24 to 8.63 ± 0.02%, and from 12.98 ± 0.71 to 33.19 ± 1.50%, respectively. In addition, DPPH radical-scavenging capacity ranged from 1.61 ± 0.10 to 16.04 ± 2.30 mg ascorbic acid equivalent per g of dried extract, and Trolox equivalent antioxidant capacity ranged from 3.03 ± 0.86 to 42.24 ± 0.72 mg Trolox equivalent per g of dried extract. Phenolic content was correlated with both DPPH radical-scavenging capacity (r = 0.63) and Trolox equivalent antioxidant capacity (r = 0.29). Nine accessions (IT170290, IT170291, IT170270, IT170276, IT170379, IT170386, IT170388, IT170418, and IT170340) exhibited early maturity, compact and erect growth habits, and above average antioxidant activities and phenolic and protein contents. In addition, accessions IT170290 and IT170291 were especially promising pigeon pea germplasms to grow, owing to various favorable characteristics (e.g., high yield and dietary fiber content). Hence, these accessions could be useful cultivars to the Republic of Korea if considered in future agricultural systems.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.7
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• pp.974-981
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• 2002

Faba beans (vicia faba) (FB) and lupin seeds (Lupinus Albus) (LS) were dry roasted at three temperatures (110, 130, $150^{\circ}C$) for 15, 30 or 45 min to determine the effects of dry roasting on rumen degradation of crude protein and starch free organic matter ($^{PSF}OM$). Rumen degradation characteristics of $^{PSF}OM$ were determined by the nylon bag incubation technique in dairy cows fed 60% hay and 40% concentrate. Measured characteristics of $^{PSF}OM$ were undegradable fraction (U), degradable fraction (D), soluble fraction (S), lag time (T0), and the rate of degradation (Kd). Based on the measured characteristics, rumen availability ($RA^{PSF}OM$) and bypass $^{PSF}OM$ ($B^{PSF}OM$) were calculated. Dry roasting did not have a greater impact on rumen degradation characteristics of $^{PSF}OM$ (p>0.05). S varied from 32.1 (raw) to 30.0, 27.8, 30.8% (LS) and 15.4 (raw) to 14.4, 20.8, 20.9% (FB); D varied from 65.4 (raw) to 66.3, 66.9, 55.9% (LS) and 54.9 (raw) to 55.0, 51.0, 64.7% (FB); U varied from 2.6 (raw) to 7.3, 7.0, 7.7% (LS) and 29.7 (raw) to 30.6, 28.2, 14.4% (FB); Kd varied from 6.0 (raw) to 7.3, 7.0, 7.7% (LS) and 22.4 (raw) to 24.4, 21.1, 7.9% (FB); $B^{PSF}OM$ varied from 35.5 (raw) to 33.8, 36.6, 38.2% (LS) and 41.3 (raw) to 41.5, 39.7, 47.6% (FB) at 110, 130 and $150^{\circ}C$, respectively. Therefore dry roasting did not significantly affect $RA^{PSF}OM$, which were 353.7, 367.9, 349.6, 336.9 (g/kg DM) (LS) and 12.82, 127.0, 133.7, 117.1 (g/kg DM) (FB) at 110, 130 and $150^{\circ}C$, respectively. These results alone with our previously published reports indicate dry roasting had the differently affected pattern of rumen degradation characteristics of various components in LS and FB. It strongly increased bypass crude protein (BCP) and moderately increased starch (BST) with increasing temperature and time but least affected $^{PSF}OM$. Such desirable degradation patterns in dry roasted LS and FB might be beneficial to the high yielding cows which could use more dry roasted $^{PSF}OM$ as an energy source for microbial protein synthesized in the rumen and absorb more amino acids and glucose in the small intestine.