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Protoplast Fusion of Nicotiana glauca and Solanum tuberosum Using Selectable Marker Genes (표식유전자를 이용한 담배와 감자의 원형질체 융합)

  • Park, Tae-Eun;Chung, Hae-Joun
    • The Journal of Natural Sciences
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    • v.4
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    • pp.103-142
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    • 1991
  • These studies were carried out to select somatic hybrid using selectable marker genes of Nicotiana glauca transformed by NPTII gene and Solanum tuberosum transformed by T- DNA, and to study characteristics of transformant. The results are summarized as follows. 1. Crown gall tumors and hairy roots were formed on potato tuber disc infected by A. tumefaciens Ach5 and A. rhizogenes ATCC15834. These tumors and roots could be grown on the phytohormone free media. 2. Callus formation from hairy root was prompted on the medium containing 2, 4 D 2mg/I with casein hydrolysate lg/l. 3. The survival ratio of crown gall tumor callus derived from potato increased on the medium containing the activated charcoal 0. 5-2. 0mg/I because of the preventions on the other hand, hairy roots were necrosis on the same medium. 4. Callus derived from hairy root were excellently grown for a short time by suspension culture on liquid medium containing 2, 4-D 2mg/I and casein hydrolysate lg/l. 5. The binary vector pGA643 was mobilized from E. coli MC1000 into wild type Agrobacteriurn tumefaciens Ach5, A. tumefaciens $A_4T$ and disarmed A. tuniefaciens LBA4404 using a triparental mating method with E. ccli HB1O1/pRK2013. Transconjugants were obtained on the minimal media containing tetracycline and kanamycin. pGA643 vectors were confirmed by electrophoresis on 0.7% agarose gel. 6. Kanamycin resistant calli were selected on the media supplemented with 2, 4-D 0.5mg/1 and kanamycin $100\mug$/ml after co- cultivating with tobacco stem explants and A. tumefaciens LBA4404/pGA643, and selected calli propagated on the same medium. 7. The multiple shoots were regenerated from kanamycin resistant calli on the MS medium containing BA 2mg/l. 8. Leaf segments of transformed shoot were able to grow vigorusly on the medium supplemented with high concentration of kanamycin $1000\mug$/ml. 9. Kanamycin resistant shoots were rooting and elongated on medium containing kanamycin $100\mug$/ml, but normal shoot were not. 10. For the production of protoplast from potato calli transformed by T-DNA and mesophyll tissue transformed by NPTII gene, the former was isolated in the enzyme mixture of 2.0% celluase Onozuka R-10, 1.0% dricelase, 1.0% macerozyme. and 0.5M mannitol, the latter was isolated in the enzyme mixture 1.0% Celluase Onozuka R-10, 0.3% macerozyme, and 0.7M mannitol. 11. The optimal concentrationn of mannitol in the enzyme mixture for high protoplast yield was 0.8M at both transformed tobacco mesophyll and potato callus. The viabilities of protoplast were shown above 90%, respectively. 12. Both tobacco mesophyll and potato callus protoplasts were fused by using PEG solution. Cell walls were regenerated on hormone free media supplemented with kanamycin after 5 days, and colonies were observed after 4 weeks culture.

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Palaeomagnetism of the Cretaceous Yuchon Group in Kosong Area, Southern Kyongsang Basin (경상분지 남단 고성지역의 백악기 유천층군에 대한 고자기 연구)

  • Kang, Hee-Cheol;Kim, In-Soo
    • Journal of the Korean earth science society
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    • v.21 no.6
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    • pp.663-674
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    • 2000
  • A total of 165 independently oriented core samples were collected from 19 Cretaceous Yuchon Group sites in Kosong area, the southernmost part of the Miryang subbasin of the Kyongsang Basin in southern Korea. Stepwise AF and thermal cleaning revealed antipodal ChRM from 95 samples from 14 sites. Mean ChRM direction is d=26.0$^{\circ}$, i=49.4$^{\circ}$ (${\alpha}_{95}$=8.2$^{\circ}$, k=24.5, n= 14) before bedding correction and d=28.1$^{\circ}$, i=54.2$^{\circ}$ (${\alpha}_{95}$=4.8$^{\circ}$, k=70.6, n= 14) after bedding correction. A 2.88-fold increase of the precession parameter k by bedding correction indicates pre-folding age of the ChRM with 99% confidence level. Palaeomagnetic pole position calculated from the mean ChRM is 67.0$^{\circ}$N, 210.6$^{\circ}$E (dp=4.7$^{\circ}$, dm=6.7$^{\circ}$), which is significantly different neither from the poles of other part of the Kyongsang Basin nor those of Eurasia including SCB and NCB. This suggests stable relative position of the study area with regard to other parts of the Kyongsang Basin as well as to Eurasia continent since Cretaceous. Three ploarity reversals in the Kosong Formation in addition to the coexistence of normal and reversed polarities in the overlying Andesites and Welded Tuff suggest, in reference to the worldwide geomagnetic polarity time scale, an Albian to Maastrichtian (polarity chron 32r-31r) age of the Yuchon Group of the study area. An alleged hypothesis of stratigraphical correspondence between the Kosong Formation in the study area and the Tadaepo Formation in Pusan area is, however, not tenable: Not only because the latter shows a short reverse polarity only in its lowest part of the sequence but also because the Andesites overlying it is wholly normally magnetized, in contrast to the frequent reverals in the case of both the Kosong Formation and Andesites above it.

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Studies on the Inheritance of Agronomic Characteristics in Upland Cotton Varieties (Gossypium hirsutum L.) in Korea (육지면품종의 유용형질의 유전에 관한 연구)

  • Bang-Myung Kae
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.21 no.2
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    • pp.281-313
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    • 1976
  • To obtain fundamental informations on cotton breeding efficiences for Korea, individual genetic relationships and interrelationships between the agronomic characteristics of Upland cotton were investigated. These experiments were couducted at the Mokpo Branch Station $(34^{\circ}48'N, $ $126^{\circ}23'E$ and altitude of 10m above sea level) from 1969 through 1972. Heterosis, combining ability, dominance and recessive gene action, genetic variance, and phenotypic and genotypic correlation were investigated by $F_1'S$ from an 11-parent partial diallel cross and the segregating $F_2$ and $F_3$ populations of the cross Paymaster times Heujueusseo Trice. The following points resulted from this study, 1. Heteroses for number of bolls per plant and lint yield were significant at 27, 84% and 37.26%, respectively. No other character had significant heteroses. 2. The GCA estimates for all studied characteristics were higher than the SCA estimates. Varieties with high GCA effects were Suwon 1 for earliness, Paymaster and Arijona for high lint percent, and Arijona for long fiber, etc, 3. SCA estimates for lint yield varied widely in crosses with Mokpo 4, Mokpo 6 and Heujueusseo Trice. Those crosses with the highest SCA effects were combinations with large characteristics differences, Example of these crosses are Mokpo 4 times Acala 1517W, Mokpo 4 times D. P. L. and Heujueusseo Trice aud Paymaster. 4. Early-maturing varieties were completely dominant to late-maturing varieties in some combinations while other crosses gave intermediate phenotypes. These results suggest additive genetic action by multi-genes. Heujueusseo Trice, Mokpo 6, and Suwon 1 showed highest degree of dominance for earliness. 5. There were no significant trends for inheritance of weight of boll and 100 seeds weight. 6. Long staple was partially to completely dominant to short staple. Though there were single gene ratios the rate of dominance decreased in the $F_2$ and $F_3$ populations in the cross between the long staple variety Paymaster and the short staple variety Heujueusseo Trice. Diallel cross $F_1$ hybrids showed complicated allelic gene action for staple length. Various dominance degree were shown by varieties. 7. Number of bolls per plant indicated strong over-dominance and small non-allelic additive gene action. 8. Lint Yield was characterized by over-dominance and by multiple non-allelic-gene action. High-yielding varieties were dominant to low-yielding ones. However, the low-yielding variety Heujueusseo Trice showed over-dominance, indicating different reactions according to the varieties and combinations. 9. Broad sense heritability for days to flowering was 34-39% while narrow sense heritability was 11%. Large variations of individual plants caused by Korean climatic conditions cause this situation. Heritability estimates for weight of boll was 30% for broad sense and 22% for narrow sense. 10. Heritability estimates for staple length and lint percent were very high suggesting strong selection effects. 11. Narrow sense heritability estimates for number of bolls per plant was 30% in the diallel cross $F_1$ hybrids and 36% in the $F_2$ population of the special cross. Broad sense heritability was estimated at 67% suggesting that. 12. Heritability estimates for lint yield was low due to high over-dominance in the diallel cross $F_1$ hybrids. Heritability estimates for yield was low in the $F_1$ hybrids but high in the $F_2$ and $F_3$ populations. 13. Phenotypic and genotypic correlations between lint percent and days to flowering and between staple length and days to flowering were high in the $F_1, $ $F_2$ and $F_3$ populations. Late-maturing varieties and individuals had long staple and high lint percent in general. As the correlation between days to flowering and lint yield was extremely low, the two traits were considered independent of each other. Days to flowering and number of bolls per plant were negatively correlated in the $F_3$ population, indicating early-maturing individual plants with many bolls may be readily selected. 14. Phenotypic and genotypic correlations between lint percent and staple length were high in $F_1, $ $F_2$ and $F_3$ populations. Accordingly, long staple varieties were high in lint percent. It was recognized that lint yield and lint percent were positively correlated in the diallel cross $F_1$ hybrids, and lint percent and staple length were positively correlated in the $F_2$ population, indicating that lint percent and staple length affect lint yield. 15. Lint yield was significantly and positively phenotypically correlated with number of bolls per plant in $F_1, $ $F_2$ and $F_3$ populations. A high genotypic correlation was also noted indicating a close genetic relationship. The selection efficiencies for a high-yielding variety can be increased when individual plants with many bolls are selected in later generations. The selection efficiencies for good fiber quality can be enhanced when individuals with long staple and high lint percent are selected in early generations.

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Studies on the Anther Culture of Prunus mume S. et Z. and the Other Three Species (Prunus mume S. et Z. 외(外) 삼종(三種)의 약배양(葯培養)에 관(關)한 연구(硏究))

  • Kim, Jai Saing
    • Journal of Korean Society of Forest Science
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    • v.31 no.1
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    • pp.1-7
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    • 1976
  • When haploid plant would be appeared by the anther culture, the large quantity of young plant multiplied maternal inheritance and the same pure line rapidly in the short length of time, which will be effected to cut down much expences efforts and time for the production of seeds or seedlings. Therefore, the development of the technique for this would be much profited in the country industry. In the late of a few years studies were early attempted in this field, but up this time there were a few success of plants only and none of perennial plant. In this status of the country condition required earnestly for the development of the green industry, this researcher attempted to culture the anther of late uninucleate microspore or early binucleate microspore of the Prunus mume and other three psecies, economic trees estimated specially economic, on the place of Modified Murashige and Skoog's medium supliment with Kinetine, 2.4-D, and N.A.A for inducing haploid plants. The obtained results were as follows: 1. 2,000 anthers were cultured and there were shown that 2N callus in Prunus mume had 82 as 4.1%, 2N callus in Prunus tomentosa 15 as 0.8%, 2 N callus in Prunus salisina 75 as 4% 2. N callus had shown 40 as 2% from Prunus armeniaca var. ansu only, and the other trees showed all 2N callus. 3. Callus had appeared in every tree but 2N callus appeared was all filaments and there showed from only connective tissue N callus appeared was all from anther locule inside. 4. Then Prunus armeniaca var. ansu only was not callus of somatic anther tissue origin, but as there was callus origined from microspore which was changed in to swollen microspores or polynucleate microspores, it was certained to need haploid plant.

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Volume Rendering System of e-Science Electron Microscopy using Grid (Gird를 이용한 e-사이언스 전자현미경 볼륨 랜더링 시스템)

  • Jeong, Won-Gu;Jeong, Jong-Man;Lee, Ho;Choe, Sang-Su;Ahn, Young-heon;Hur, Man-Hoi;Kim, Jay;Kim, Eunsung;Jung, Im Y.;Yeom, Heon Y.;Cho, Kum Won;Kweon, Hee-Seok
    • Proceedings of the Korea Contents Association Conference
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    • 2007.11a
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    • pp.560-564
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    • 2007
  • Korea Basic Science Institute(KBSI) has three general electron microscopes including High Voltage Electron Microscope(HVEM) which is the only one in Korea. Observed images through an electron microscope are what they are tilted by each step and saved, offering the more better circumstances for observers, a reconstruction to 3D could be a essential process. In this process, a warping method decreases distortions maximumly of avoided parts of a camera's focus. All these image treatment processes and 3D reconstruction processes are based on an accompaniment of a highly efficient computer, a number of Grid Node Personal computers share this process in a short time and dispose of it. Grid Node Personal computers' purpose is to make an owner can share different each other and various computing resources efficiently and also Grid Node Personal computers is applying to solve problems like a role scheduling needed for a constructing system, a resource management, a security, a capacity measurement, a condition monitoring and so on. Grid Node Personal computers accomplish roles of a highly efficient computer that general individuals felt hard to use, moreover, a image treatment using the warping method becomes a foundation for reconstructing to more closer shape with an real object of observation. Construction of the electron microscope volume 랜더링 system based on Grid Node Personal computer through the warping process can offer more convenient and speedy experiment circumstances to observers, and makes them meet with experiment outcome that is similar to real shapes and is easy to understand.

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Dark Fermentative Hydrogen Production using the Wastewater Generated from Food Waste Recycling Facilities (혐기 발효 공정을 통한 음식물류 폐기물 탈리액으로부터 수소 생산)

  • Kim, Dong-Hoon;Lee, Mo-Kwon;Lim, So-Young;Kim, Mi-Sun
    • Transactions of the Korean hydrogen and new energy society
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    • v.22 no.3
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    • pp.326-332
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    • 2011
  • The authors examined the effects of operating parameters on the $H_2$ production by dark fermentation of the wastewater generated from food waste recycling facilities, in short "food waste wastewater (FWW)". Central composite design based response surface methodology was applied to analyze the effect of initial pH (5.5-8.5) and substrate concentration (2-20 g Carbo. COD/L) on $H_2$ production. The experiment was conducted under mesophilic ($35^{\circ}C$) condition and a heat-treated ($90^{\circ}C$ for 20min)anaerobic digester sludge was used as a seeding source. Although there was a little difference in carbohydrate removal, $H_2$ yield was largely affected by the experimental conditions, from 0.38 to 1.77 mol $H_2$/mol $hexose_{added}$. By applying regression analysis, $H_2$ yield was well fitted based on the coded value to a second order polynomial equation (p = 0.0243): Y = $1.78-0.17X_1+0.30X_2+0.37X_1X_2-0.29X_1{^2}-0.35X_2{^2}$, where $X_1$, $X_2$, and Y are pH, substrate concentration (g Carbo. COD/L), and hydrogen yield (mol $H_2$/mol $hexose_{added}$), respectively. The 2-D response surface clearly showed a high inter-dependency between initial pH and substrate concentration, and the role of these two factors was to control the pH during fermentation. According to the statistical optimization, the optimum condition of initial pH and substrate concentration were 7.0 and 13.4 g Carbo. COD/L, respectively, under which predicted $H_2$ yield was 1.84 mol $H_2$/mol $hexose_{added}$. Microbial analysis using 16S rRNA PCR-DGGE showed that $Clostridium$ sp. such as $Clostridium$ $perfringens$, $Clostridium$ $sticklandii$, and $Clostridium$ $bifermentans$ were main $H_2$-producers.

DNA Polymorphisms in SREBF1 and FASN Genes Affect Fatty Acid Composition in Korean Cattle (Hanwoo)

  • Bhuiyan, M.S.A.;Yu, S.L.;Jeon, J.T.;Yoon, D.;Cho, Y.M.;Park, E.W.;Kim, N.K.;Kim, K.S.;Lee, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.6
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    • pp.765-773
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    • 2009
  • Sterol regulatory element binding factor 1 (SREBF1) and fatty acid synthase (FASN) genes play an important role in the biosynthesis of fatty acids and cholesterol, and in lipid metabolism. This study used polymorphisms in the intron 5 of bovine SREBF1 and in the thioesterase (TE) domain of FASN genes to evaluate their associations with beef fatty acid composition. A previously identified 84-bp indel (L: insertion/long type and S: deletion/short type) of the SREBF1 gene in Korean cattle had significant associations with the concentration of stearic (C18:0), linoleic (C18:2) and polyunsaturated fatty acids (PUFA). The stearic acid concentration was 6.30% lower in the SS than the LL genotype (p<0.05), but the linoleic and PUFA contents were 11.06% and 12.20% higher in SS compared to LL (p<0.05). Based on the sequence analysis, five single nucleotide polymorphisms (SNPs) g.17924G>A, g.18043C>T, g.18440G>A, g.18529G>A and g.18663C>T in the TE domain of the FASN gene were identified among the different cattle breeds studied. Among these, only g.17924 G>A and g.18663C>T SNPs were segregating in the Hanwoo population. The g.17924G>A SNP is a non-synonymous mutation (thr2264ala) and was significantly associated with the contents of palmitic (C16:0) and oleic acid (C18:1). The oleic acid concentration was 3.18% and 2.79% higher in Hanwoo with the GG genotype than the AA and AG genotypes, respectively (p<0.05), whereas the GG genotype had 3.8% and 4.01% lower palmitic acid than in those cattle with genotype AA and AG, respectively (p<0.05). Tissue expression data showed that SREBFI and FASN genes were expressed in a variety of tissues though they were expressed preferentially in different muscle tissues. In conclusion, the 84-bp indel of SREBF1 and g.17924G>A SNP of the FASN gene can be used as DNA markers to select Hanwoo breeding stock for fatty acid composition.

Development of Biotelemetry Method by Combining the SSBL Method and the Pinger Synchronizing Method (1) - Design and production of system - (SSBL 방식과 핑거동기 방식을 조합한 바이오텔레메터리 방식의 개발 (1) -시스템의 설계 및 제작 -)

  • 박주삼;고탁창언
    • Journal of the Korean Society of Fisheries and Ocean Technology
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    • v.39 no.3
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    • pp.218-229
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    • 2003
  • A new biotelemetry method that the installation and the treatment of equipment is convenient and the instantaneously detailed location of the fish attached the pinger is able to track comparatively easily was developed. The receiving system in this biotelemetry method was advanced for track the detailed behavior of the fish by the miniature tracking pinger, because it was a burden to fish to add the pinger with the water temperature and the pressure sensor. By combining of the super short base line (SSBL) method to detect the direction of pinger and the pinger synchronizing method to measure the range from receiving transducer to pinger, the three dimensional locations of fish to the receiving transducer is gotten instantaneously. The receiving system is devised to realize the high precision or wide detection range by application of the basic design method for receiving system of biotelemetry developed by the present authors and the hydrophone array configuration. The measurement distance error in the pinger synchronizing method is minimized through the correction of which the deviation of transmission pluse period of pinger is caused by changing water temperature. A prototype system which is able to track the instantaneously detailed location of the fish by the SSBL and pinger synchronizing biotelemetry (SPB) method was produced.

Evaluation of Potential Biomarkers for Thioacetamide-induced Hepatotoxicity using siRNA

  • Kang, Jin-Seok;Yum, Young-Na;Han, Eui-Sik;Kim, Joo-Hwan;Lee, Eun-Mi;Ryu, Doug-Young;Kim, Young-Hee;Yang, Sung-Hee;Kim, Seung-Hee;Park, Sue-Nie
    • Biomolecules & Therapeutics
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    • v.16 no.3
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    • pp.197-202
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    • 2008
  • In our previous publication we compared the gene expression profiles on hepatotoxicants exposure to assess the comparability between in vivo and in vitro test systems. We investigated global gene expression from both mouse liver and mouse hepatic cell line treated with thioacetamide (TAA) and identified several common genes. In this study, we selected genes to validate them as potential biomarkers for hepatotoxicity on the relevance of in vitro and in vivo system. Three up-regulated, aquaporin 8 (Aqp8), glutathione peroxidase 1 (Gpx1), succinate-CoA ligase, GDP-forming, alpha subunit (Suclg1) and two down-regulated, DnaJ (Hsp40) homolog subfamily C member 5 (Dnajc5) and tumor protein D52 (Tpd52) genes were tested for their effects in vitro. For characterization of gene function, short interfering RNA (siRNA) for each gene was synthesized and transfected in mouse hepatic cell line, BNL CL.2. Cell viability, mRNA expression level and morphological alterations were investigated. We confirmed siRNA transfection against selected five genes induced down-regulation of respective mRNA expression. siRNA transfection in general decreased cell viability in different degrees and induced morphological changes such as membrane thickening and alterations of intracellular structures. This suggests that these genes could be associated with TAA-induced toxicity. Furthermore, these genes may be used in the investigation of hepatotoxicity for better understanding of its mechanism.

Metabolic Engineering of the Brassinosteroid Biosynthetic Pathways (Brassinosteroid의 대사공학)

  • LEE Mi-Ock;SONG Ki-Hong;LEE Hyun-Kyung;JUNG Ji-Yoon;CHOE Vit-Nary;CHOE Sunghw
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2002.04a
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    • pp.69-75
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    • 2002
  • Sterols play two major roles in plants: a bulk component in biological membranes and precursors of plant steroid hormones. Physiological effects of plant steroids, brassinosteroids (BRs), include cell elongation, cell division, stress tolerance, and senescence acceleration. Arabidopsis mutants that carry genetic defects in BR biosynthesis or its signaling display characteristic phenotypes, such as short robust inflorescences, dark-green round leaves, and sterility. Currently there are more than 100 dwarf mutants representing 7 genetic loci in Arabidopsis. Mutants of 6 loci, dwf1/dim1/cbb1, cpd/dwf3, dwf4, dwf5, det2/dwf6, dwf7 are rescued by exogenous application of BRs, whereas bri1/dwf2 shares phenotypes with the above 6 loci but are resistant to BRs. These suggest that the 6 loci are defective in BR biosynthesis, and the one locus is in BR signaling. Biochemical analyses, such as intermediate feeding tests, examining the levels of endogenous BR, and molecular cloning of the genes revealed that dwf7, dwf5, and dwf1 are defective in the three consecutive steps of sterol biosynthesis, from episterol to campesterol via 5-dehydroepisterol. Similarly, det2/dwf6, dwf4, and cpd/dwf3 were shown to be blocked in $D^4$ reduction, 22a-hydroxylation, and 23 a-hydroxylation, respectively. A signaling mutant bri1/dwf2 carries mutations in a Leucine-rich repeat receptor kinase. Interestingly, the bri1 mutant was shown to accumulate significant amount of BRs, suggesting that signaling and biosynthesis are dynamically coupled in Arabidopsis. Thus It is likely that transgenic plants over-expressing the rate-limiting step enzyme DWF4 as well as blocking its use by BRI1 could dramatically increase the biosynthetic yield of BRs. When applied industrially, BRs will boost new sector of plant biotechnology because of its potential use as a precursor of human steroid hormones, a novel lead compound for cholesterol-lowering effects, and a various application in plant protection.

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