• Title/Summary/Keyword: short-hair seed

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Analysis of Seed Hair Formation Related Genes by EST Profiling in Carrot (Daucus carota var. sativa) (EST profiling을 통한 당근(Daucus carota var. sativa)의 종모 형성에 관련된 유전자 분석)

  • Hwang, Eun-Mi;Oh, Gyu-Dong;Shim, Eun-Jo;Jeon, Sang-Jin;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.28 no.6
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    • pp.1039-1050
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    • 2010
  • Carrot is one of the useful crops used abundantly in cooking in Western as well as Asia regions such as China and Korea. However, seed coats have hairs which should be removed to increase germination rate. Furthermore, because of seed hairs, farmers face several additional losses, such as time consumption, manpower, capital and so on, for seed handling. To prevent these problems, study of gene related hair formation using short-hair seed lines is required. We analyzed genes related to hair formation from seed through expressed sequenced tag (EST) profiling, based on the fact that the development of carrot seed hair is related to cellulose synthesis pathway in secondary cell wall synthesis stage. To study the gene expression related to hair formation of the carrot seed, a cDNA library was constructed by using the early maturation stage of the short-hair line (659-1) and hairy seed line (677-14). In short-hair (659-1) and hairy seed (677-14) lines, results from of EST profiling through BLASTX search analysis using the NCBI database showed that 172 and 224 unigenes had significant homology with known protein sequences, whereas 233 and 192 unigenes were not, respectively. All ESTs were grouped into 16 categories according to their putative functions. Twenty nine unigenes among all ESTs were considered to be genes regulating seed hair development from cellulose synthesis pathway during secondary cell wall synthesis stage; in results, 14 unigenes related to seed hair development were found only in hairy seed line.

Development of SNP Molecular Markers Related to Seed-hair Characteristic Based on EST Sequences in Carrot (당근 EST 염기서열을 이용한 종자모 형질 관련 SNP 분자표지 개발)

  • Oh, Gyu-Dong;Shim, Eun-Jo;Jun, Sang-Jin;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.31 no.1
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    • pp.80-88
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    • 2013
  • Carrot (Daucus carota L. var. sativa) is one of the most extensively used vegetable crops in the world and a significant source of nutrient because of its high content of ${\beta}$-carotene, well known as the precursor of vitamin A carotenoid. However, seed-hairs generated and elongated from the epidermal cell of seeds inhibit absorption and germination by various factors such as carotol and so on. Accordingly, mechanical hair removal process is essential before commercialization of carrot seeds. Because of this process, producers will have additional losses such as time consuming, manpower, capital and so on. Furthermore, physical damage of seeds causes irregular germination rate. To overcome such cumbersome weaknesses, new breeding program for developing hairless-seed carrot cultivar has been needed and studies for molecular markers related to seed-hair characteristic is needed for a new breeding program. Therefore, in this study, cDNA libraries from seeds of short-hair seed phenotype CT-SMR 616 OP 659-1 line, hairy-seed phenotype CT-SMR 616 OP 677-14 line and short-hair seed phenotype CT-ATR 615 OP 666-13 line, hairy-seed phenotype CT-ATR 615 OP 671-9 were constructed, respectively. Furthermore, 1,248 ESTs in each line, total 4,992 ESTs were sequenced. As a result, 19 SNP sites and 14 SNP sites in each of 2 combinations were confirmed by analyzing these EST sequences from short-hair and hairy-seed lines. Then we designed SNP primer sets from EST sequences of SNP sites for high resolution melting (HRM) analysis. Designed HRM primers were analyzed using hairy seed phenotype CT-SMR 616 OP 1040 line and short-hair seed phenotype CT-SMR 616 OP 1024, 1025, 1026 lines. One set of HRM primers showed specific difference between the melting curves of hairy and short-hair seed phenotype lines. Based on this result, allele-specific (AS) PCR primers were designed for easier selection between hairy-seed carrot and hairless seed carrot. These results of HRM and AS-PCR are expected to be useful in breeding of hairless seed carrot cultivar as a molecular marker.

Development of RAPD-SCAR Molecular Marker Related to Seed-hair Characteristic in Carrot (당근(Daucus carota var. sativa) 종자모 형질 관련 RAPD-SCAR 분자표지 개발)

  • Shim, Eun-Jo;Park, Sung-Kwan;Oh, Gyu-Dong;Jun, Sang-Jin;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.31 no.6
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    • pp.756-763
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    • 2013
  • Mechanical hair removal of carrot seed causes seed injuries and suppresses the germination in carrot cultivation. This study was performed to develop molecular markers for breeding high quality cultivars with short-hair seed. To meet this objective, random amplified polymorphic DNA (RAPD)-sequence characterized amplified region (SCAR) markers specifically linked to seed-hair characteristic were identified using CT-SMR 616 OP 389-1 line with short-haired seed and CT-SMR 616 OP 616-33 line with long-haired seed, bred by self-pollination for 6 years from 2008 to 2013, as parents. After seed hair lengths of these lines were analyzed using microscope, next generations were advanced and compared with the molecular markers polymorphism. From RAPD analysis using fixed lines in 2011, twelve RAPD primers showing polymorphic bands specific between the two lines were identified from 80 random primers. To develop RAPD-SACR marker, SCAR primers were designed based on sequence analysis of these specific RAPD bands and more than three combinations of primers were tested. As a result, it was found that the $SCA2_{1.2}$ amplified single polymorphic band from short-haired seed line. To confirm this result, $SCA2_{1.2}$ marker was retested by applying to the 2012 and 2013 progenies. Finally, it was concluded that the developed $SCA2_{1.2}$ marker distinguished short-haired line from long-haired seed line. Therefore, SCAR marker, $SCA2_{1.2}$ is expected to be utilized for breeding of the short-haired seed cultivars.

Construction of cDNA Library and EST Analysis Related to Seed-hair Characteristics in Carrot (당근 종모 형질 관련 cDNA Library 작성 및 EST 분석)

  • Oh, Gyu-Dong;Shim, Eun-Jo;Jun, Sang-Jin;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.31 no.6
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    • pp.782-789
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    • 2013
  • Carrot (Daucus carota L. var. sativa) is one of the most widely used crops in the world and is nutritionally important crop. However, seed-hair which is generated in epidermal cell of seeds causes the difficulty of the seedling process, because of the seed germination and absorption inhibitions. For these reasons, carrot seeds are commercialized after mechanical hair removal process. However, in this process, various damage and seed loss occur and breeding of hairless-seed carrot cultivar is needed to overcome these various weaknesses and additional seed production costs. In this study, cDNA libraries using 2 combinations, which were composed of short-hair seed CT-ATR 615 OP 666-13 & long-hair seed CT-ATR 615 OP 671-9, and short-hair seed CT-SMR 616 OP 659-1 & long-hair seed CT-SMR 616 OP 677-14, were constructed and EST sequences of each individuals were analyzed to reveal carrot seed-hair characteristics. Firstly, analyzed EST sequences were classified into FunCat functional categories. As a result, significant differences have been identified in metabolism category, protein folding and stabilization, protein binding, C-compound binding category from both of two combinations. Secondly, several candidate EST sequences related to seed trichome differentiation and cellulose biosynthetic process were selected based on GO data of EST sequences. These differences based on FunCat categories and candidate EST obtained by GO data analysis are thought to be involved in the formation of carrot seed hair. Finally, 741 SSR sites and 33 SNP sites were identified from analyzed EST sequences of two combinations. Then we designed SNP and SSR primer sets to develop molecular markers. These molecular markers will be used for classification of carrot cultivars and study seed-hair characteristic.

EST Profiling for Seed-hair Characteristic and Development of EST-SSR and SNP Markers in Carrot (당근 종모 형질 관련 EST profiling과 이를 이용한 EST-SSR 및 SNP 마커 개발)

  • Oh, Gyu-Dong;Hwang, Eun-Mi;Shim, Eun-Jo;Jeon, Sang-Jin;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.28 no.6
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    • pp.1025-1038
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    • 2010
  • Carrot ($Daucus$ $carota$ L. var. $sativa$) is one of the most widely used crops in the world. Moreover it is an important crop because of its high content of ${\beta}$-carotene, well-known as the precursor of vitamin A carotenoid. However, seed-hair which is generated in epidermal cell of seeds inhibits absorption and germination. For that reason, carrot seeds are commercialized after mechanical hair removal process. To overcome such cumbersome weaknesses, new breeding program for developing hairless-seed carrot cultivar has been needed. Therefore, in this study, cDNA libraries from seeds of short-hair seed phenotype CT-ATR615 OP 666-13line and hairy seed CT-ATR615 OP-CK1-9 line were constructed and expression patterns related to generation of seed-hair were analyzed by comparison of EST sequences. Differential EST sequence results between two lines were classified into FunCat functional categories based on the results of BlastX search. Higher expression quantities belonging to metabolic category were shown on short-hair seed line than hairy-seed one. Differential expression quantities between those two lines in the protein folding and stabilization, subcellular localization categories were supposed to contribute variously on the generation of seed-hair. We confirmed 50 and 59 SSR sites, and 2 SNP sites by analyzing EST sequences in two lines; thereafter, we designed SNP and SSR primer sets from these EST sequence information as a molecular marker. These markers are thought to be used in research of molecular markers for classification of carrot family and related to various traits, as well as seed-hair characteristic.

Characteristic and Application Under the Sericulture of Subtropical Zones Mulberry Adapted Itself to the Field Cultivation (노지재배(露地栽培)에 적응(適應)한 아열대산(亞熱帶産) 뽕나무의 특성(特性)과 양잠(養蠶)에서의 응용(應用))

  • Seok Young-Seek;Park Sang-Jo;An Sin-Hun;Han Sang-Mi;Yeo Joo-Hong;Han Myung-Sae
    • Journal of Sericultural and Entomological Science
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    • v.47 no.2
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    • pp.68-77
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    • 2005
  • A characteristic of subtropical zones region MK-T2 compares with an gaeryangppong, and the 9-10 schedule the times when a leaf blooms to are fast, and ratio that a branch edge by the colds becomes lean showed 5.7%, and a growth of the new branch which went out delivers 67.2 cm, mulberry loaves of the new branch which went out, and 18.6, a form of a leaf is the 1.10 that length of a leaf grew more a bit than width of a leaf up. Thickness of a leaf is $228.2{\mu}m$, and an area is more similar than gaeryangppong as $225.6cm^2$. in plant taxonomy, the hair whom the style exists short with 0.7 mm, and go to the pistil head inside so as to be rare is distributed, and belong to Dolichostylae Pubescentes. The new branch cutting which executed without remedy processes was independent of a thickness of a case branch, and the form and 100% root was said, and an gaeryangppong compared with the fact that 10% root went out of 15 mm ideal, and was excellent very, and looked, a root went out a root the soil and water, all showed a characteristic to go out at central of a branch bases at 45% ratio. Length was 24.6 mm, and were water rate 78.8%, and mulberry of MK-T2 was carrying together sweetness and acidity to pH 4.7 while, besides, arrival was 19.21 Brix%. A larva period and pupa ratio, cocoon thickness ratio are almost similar to gaeryangppong, or weight of one cocoon, cocoon thickness, 20,002 cocoon quantity shows some results to drop, and be soft of a leaf, and feed value certifications are comparatively top-ranking. As a result of having analyzed amino acid of the 3rd day of 5th silkworm larva which bred to MK-T2, a collation absorbing an gaeryangppong went, and looked, but compared with a collation in case of tests to eat MK-T2, and looked, and the lie collations were not detected a difference at Leu, but MK-T2 tests were detected mutual almost similar amino acid creation. medical efficacy of the 3rd day of 5th silkworm larva ethanol extract which bred to MK-T2 and black results, histologic a case did not appear at HE dyeing about the kidney organization which extracted form the rats which ate a silkworm ethanol extract and dyeing all chemical organization immunity, and one step protein revelation became lower with almost unidentified levels.