This study was conducted to compare changes of total carotenoid contents and antioxidant activities in yellow waxy corns (Zea mays L.) depending on harvest time. The total carotenoid contents of yellow waxy inbred lines and hybrids increased from 19 days and then the highest content of total carotenoid contents was observed at 25 to 27 days after pollination. Thereafter, there was little change in total carotenoid contents. According to analysis total content of carotenoids on 23 days after pollination, which was the period of harvesting waxy corn, KY2 was the highest as 15.4 ㎍/g in inbred lines and KY2/KY39 was the hightest as 11.2 ㎍/g in hybrids, respectively. The total carotenoid contents of each hybrid were higher than mean that of their parent lines except for some hybrids. The total carotenoid contents of KY27/KY37 were significantly higher than those of their parental lines. Antioxidant activity of yellow waxy corn showed a tendency to increase after decrease depending on delaying harvesting time. Antioxidant activities of hybrids were higher than that of inbred lines. As a result of correlation analysis between total carotenoid contents and antioxidant activity, correlation coefficient of inbred lines and hybrids was as low as -0.12 and -0.13, respectively. When the harvest time was delayed, the lightness of yellow waxy corn decreased but the redness and yellowness increased. As a result of correlation analysis between Hunter’s Lab value and total carotenoid contents, correlation coefficient of lightness, redness and yellowness were -0.22, 0.67, 0.53, respectively.

본 연구를 통해 논, 시설재배 밭 토양, 쓰레기장, 하천 및 호수의 퇴적 토양 등 22개소에 서 분리한 총 6종의 광합성세균 중 호기 ․암 배양이 가능한 Rhodobacter sphaeroides PS-24 를 분리하였다. 형태학적 특징으로는 그람음성의 막대모양으로, 운동성이 있었다. 분리균주 의 16S rRNA 염기서열을 분석한 결과 Rhodobacter sphaeroides ATH2.4.1과 99%의 상동성을 나타내었으며, 본 연구에서 Rhodobacter spharoides PS-24로 명명하여 연구를 수행하였다. 선별균주를 modifed Van niel's yeast 배지에서 배양 후 생성된 carotenoid를 추출한 결과 12.03±0.15 mg/L의 함량이 측정되었으며, 반응표면분석법 중 Plackett burman 분석방법과 Box-Behnken 분석방법을 통해 carotenoid 생산에 영향을 미치는 요인을 분석하고 농도를 최 적화하였다. 분석결과 각각의 독립변수 yeast extract –0.4144 (1.23 g/L), Na2CO3 0.8541 (3.71 g/L)와 MgSO4 1.00 (1.00 g/L)의 농도를 선정하였으며, 이를 바탕으로 배지 조성을 최적화 한 결과 yeast extract 1.23 g, sodium acetate 1 g, NH4Cl 1.75 g, NaCl 2.5 g, K2HPO4 2 g, MgSO4 1.0 g, mono-sodium glutamate 7.5 g, Na2CO3 3.71 g, NH4Cl 3.5 g, CaCl2 0.01 g/ liter로 선정하였다. 최적배지를 대상으로 5 L, 50 L, 500 L scale-up을 진행한 결과 최종 carotenoid 는 각각 17.98 mg/L, 18.03 mg/L, 18.11 mg/L로 조사되었다. 최적배지의 경우 modified Van niel's yeast 배지보다 약 1.5배 많은 carotenoid를 생산하였으며, 대량배양을 통한 scale-up 과 정 시 carotenoid의 생산량은 크게 변화하지 않는 것으로 조사되었다. 따라서 본 연구를 바 탕으로 산업적으로 다양하게 사용되고 있는 carotenoid를 생산하는 광합성세균 Rhodobacter spharoides PS-24를 개발하였으며, 본 연구를 바탕으로 유기농축산에 사용이 가능한 기능성 미생물제제를 개발하고자 한다.

Background : Tagetes species which belong to Asteraceae show different characteristics including, bloom size, shape, color, plant size, and leaf shape. The color of Tagetes flowers ranging from white to dark orange is due to accumulation of different carotenoids, pathway intermediates, and amount of the same carotenoid. Methods and Results : The carotenoids were monitored in flower extracts from six cultivars of Tagetes that include three T. erecta cultivars, Discovery Orange (DO), Inca Orange (IO), and Inca Yellow (IY), and three T. patula cultivars, including Durango Bee (DB), Durango Yellow (DY), and Safari Red (SR) using HPLC analysis. It showed considerable differences in carotenoid composition depending on cultivars and types of carotenoids. The highest concentration of violaxanthin which represents orange color in plants was showed in IO, whereas the compound was not detected in DB, and DY. Yellow-colored cultivars such as IY, DB, and DY exhibited low levels of lutein. However, others that indicate orange color, DO, IO, and SR showed high levels of lutein. Also, similar pattern was found in the zeaxanthin measurements. α-carotene was significantly accumulated in SR compared to other cultivars. The highest amount of β-carotene was found in SR, followed by IO, IY, DO, DY and DB. Similarly, the highest and lowest amount of 9-cis-β-carotene was showed in SR and DB, respectively. Interestingly, all cultivars except SR in 13-cis-β-carotene showed the same pattern with β-carotene, but no detection indicated in SR. Conclusion : In this study, we determined the differences in carotenoid yields among six Tagetes cultivars. In total, seven carotenoids that include violaxanthin, lutein, zeaxanthin, α -carotene, β-carotene, 9-cis-β-carotene, and 13-cis-β-carotene were detected. Among them, all of the cultivars accumulated primarily lutein. In addition, contents of each carotenoid varied in these flowers depending on cultivars.

Background : Tagetes species which belong to Asteraceae show different characteristics including, bloom size, shape, color, plant size, and leaf shape. The color of Tagetes flowers ranging from white to dark orange is due to accumulation of different carotenoids, pathway intermediates, and amount of the same carotenoid. Methods and Results : The carotenoids were monitored in flower extracts from six cultivars of Tagetes that include three T. erecta cultivars, Discovery Orange (DO), Inca Orange (IO), and Inca Yellow (IY), and three T. patula cultivars, including Durango Bee (DB), Durango Yellow (DY), and Safari Red (SR) using HPLC analysis. It showed considerable differences in carotenoid composition depending on cultivars and types of carotenoids. The highest concentration of violaxanthin which represents orange color in plants was showed in IO, whereas the compound was not detected in DB, and DY. Yellow-colored cultivars such as IY, DB, and DY exhibited low levels of lutein. However, others that indicate orange color, DO, IO, and SR showed high levels of lutein. Also, similar pattern was found in the zeaxanthin measurements. α-carotene was significantly accumulated in SR compared to other cultivars. The highest amount of β-carotene was found in SR, followed by IO, IY, DO, DY and DB. Similarly, the highest and lowest amount of 9-cis-β-carotene was showed in SR and DB, respectively. Interestingly, all cultivars except SR in 13-cis-β-carotene showed the same pattern with β-carotene, but no detection indicated in SR. Conclusion : In this study, we determined the differences in carotenoid yields among six Tagetes cultivars. In total, seven carotenoids that include violaxanthin, lutein, zeaxanthin, α -carotene, β-carotene, 9-cis-β-carotene, and 13-cis-β-carotene were detected. Among them, all of the cultivars accumulated primarily lutein. In addition, contents of each carotenoid varied in these flowers depending on cultivars.

Background : Ixeris genus has been used in traditional medicines as stomachics, sedatives, and diuretics. Ixeris dentata var albiflora is a kind of perennial herbaceous plant and one of the plants of the genus Ixeris (Asteraceae). It is well-known for edible wild vegetable in Korea, China, Japan, and Mongolia. Specially, Korean has its root and young leaf with appetizing vegetable due to bitter taste. Methods and Results : We isolated 8 genes that are involved in carotenoid biosynthesis using the Illumina/Solexa HiSeq2000 platform. In this study, a full-length cDNA clone encoding phytoene synthase (IdPSY), phytoene desaturase (IdPDS), ξ-carotene desaturase (IdZDS), lycopene β-cyclase (IdLCYB), and zeaxanthin epoxidase (IdZEP) and partial-length cDNA clones encoding lycopene ε-cyclase (IdLCYE), ε-ring carotene hydroxylase (IdCHXE), and β-ring carotene hydroxylase (IdCHXB2) were identified in I. dentata. The theoretical molecular weight (MW) and isoelectric point values of 8 genes were investigated. Sequence analyses revealed that these proteins shared high identity and conserved domains with their orthologous genes. IdPSY, IdPDS, IdZDS, IdLCYB, IdCHXB2, and IdZEP were constitutively expressed in the roots, stems, leaves, and flowers of I. dentata. Conclusion : Our study on the biosynthesis of carotenoids in I. dentata will provide basic data for elucidating the contribution of carotenoids to the considerable medicinal properties of I. dentata.

영양고추시험장에서 분양 받은 고추 유전자원 523종에 대한 카로티노이드의 정성 및 정량 분석과 ASTA color 지수를 분석하였다. 개별 카로티노이드는 capsanthin, lutein, zeaxanthin, β-cryptoxanthin, α-carotene, β-carotene 등으로 동정하였다. 총 카로티노이드 함량은 0.5-84.8 mg/100g DW으로 나타났으며 ASTA color 지수는 0.7-233.1로 나타났다. 개별 카로티노이드의 평균갑과 총 카로티노이드 함량에 대한 비율은 β-carotene 5.0 mg/100g DW (24.8%) > capsanthin 5.0 mg/100g DW (24.7%) > zeaxanthin 4.1 mg/100g DW (20.4%) > β-cryptoxanthin 3.0 mg/100g DW (14.8%) > lutein 2.7 mg/100g DW (13.3%) > α-carotene 0.4 mg/100g DW (2.0%) 순으로 분석되었다. 카로티노이드 중 capsanthin의 함량이 ASTA color 지수와 높은 상관관계를 보였으며 (R2 = 0.723) capsanthin이 23.2 mg/100g DW으로 고함유된 자원 IT223599를 선발하였다. 또한 카로티노이드 크로마토그램의 패턴에 따라 고추유전자원을 7가지 그룹으로 구분하였으며 이는 고 색소 함유 품종 육종 등 다양한 산업 소재로 활용 될 수 있다.

Carotenoid isomerase (CRTISO) catalyzes the isomerization of prolycopene to all-trans-lycopene in the carotenoid biosynthetic pathway. We isolated two full-length cDNA gene, CuCRTISO and CuCRTISO-like, from Citrus unshiu. To confirm whether these two genes have the function of the carotenoid isomerase, The full-length cDNA of CuCRTISO and CuCRTISO-like gene, respectively, were fused with 35S promoter and NOS terminal region and then transformed into tomato CRTISO mutant, Tangerine, which shows orange fruit due to lack of carotenoid isomerase activity. The mature fruit color of the transgenic line expressing CuCRTISO gene changed from orange to red, which was similar to the fruit color of the tomato “Money Maker”. We also carried out HPLC analysis to detect all-trans lycopene, which is produced from prolycopene by carotenoid isomerase. In the transgenic line expressing CuCRTISO the all trans lycopene was detected from mature fruit but in the tangerine mutant several prolycopenes were detected from it. On the other hand, the transgenic line expression of CuCRTISO gene retained the orange-color fruit at the mature stage as Tangerine mutant. These studies indicate that the CuCRTISO gene has a function of carotenoid isomerase and also plays a role of it in other plant species, and that the CuCRTISO-like gene might be not enough to produce the all trans lycopene or has a another unknown function(s).

Carotenoid isomerase (CRTISO) catalyzes the isomerization of prolycopene to all-trans-lycopene in the carotenoid biosynthetic pathway. We isolated a full-length promoter region of CuCRTISO from Citrus unshiu. We determined if the promoter encoded organ-specific or developmental-specific expression, and identified possible cis-acting promoter elements. The full-length promoter and two truncated versions were fused to the β-glucuronidase (GUS) gene and transformed into Arabidopsis thaliana. Transgenic lines expressing the full-length promoter (pCiso-Prom1) and truncated promoters (pCiso-Prom2 and pCiso-Prom3) showed the same developmental and organ-specific activity. GUS expression was detected in the cotyledon and root at 5 and 10 days after germination, mature leaf, and anther. The CuCRTISO promoter contained several cis-acting elements involved in hormonal and environmental stress. Drought stress or abscisic acid treatment did not induce GUS expression in any transgenic lines. Heat stress induced GUS expression in the pCiso-Prom1 line; this promoter construct contains the heat-stress responsive element (HSE). Ethylene and cold-stress treatments induced GUS expression only in the pCiso-Prom3 line, although all transgenic lines contained the same cis-acting ethylene and low-temperature response elements. which could indicate the existence of unknown repressor element(s) in the CuCRTISO promoter. These studies indicate that CuCRTISO promoter activity is regulated in a developmental and organ-specific manner that responds to heat, cold, and ethylene. These results provide new insights into the role of cis-acting element(s) in CuCRTISO promoter activity. (This research was supported by the Basic Science Research Program of the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2010-0007627 and 2009-0094059), and by Golden Seed Project, Ministry of Agriculture, Food and Rural Affairs (MAFRA), Ministry of Oceans and Fisheries (MOF), Rural Development Administration (RDA) and Korea Forest Service (KFS), Republic of Korea)

Carotenoids of squash play an important role in human health by acting as sources of provitamin A or as protective antioxidants. Among the 60 accessions of squash germplasm, fluorescent yellow and yellow types of flesh color got the highest count, followed by the orange, whitish yellow and greenish yellow. The redness and yellowness values of the flesh powder ranged from -2.45 to 86.09 and from 13.77 to 39.80, respectively. While the lightness and the total color difference values of flesh color varied from 67.64 to 86.09 and from 19.77 to 51.79, respectively. Colorimetric values of redness and yellowness showed positive correlation, and the correlation coefficient (r) was as high as 0.7386. The five accessions represented each flesh color types, IT195043 (orange), IT136696 (fluorescent yellow), IT186365 (yellow), IT137963 (whitish yellow), and IT180449 (greenish yellow). The total amount of carotenoid contents was in the order of orange color (104.64 mg/100 g), greenish yellow color (70.82 mg/100 g), fluorescent yellow color (32.41 mg/100 g), yellow color (8.73 mg/100 g), and whitish yellow color (4.73 mg/ 100 g). Both lutein and β-carotene were the predominant pigments of carotenoids, and lycopene was only separated and identified in the orange color flesh. According to the results, colorimetric analysis can aid breeders interested in increasing carotenoid content in squash, which could be accurately measured using a simple, reliable, and cost- and labor-efficient method for the evaluation of carotenoid pigments.

비타민A 강화벼의 복수세대에 대한 후대안정성을 Southern blot과 PCR로 분석한 결과, 비타민A 강화벼의 PAC T3～T6 세대에서는 도입된 모든 유전자들이 안정적으로 도입되어 있으며, backbone DNA는 비타민A 강화벼에 삽입되지 않았음을 확인하였다. 선발 마커로 도입된 PAT 단백질의 발현 분석 결과에서도 PAC T3～T6 복수세대에서 생육시기별 부위별로 안정적으로 발현됨을 입증하였으며, 최종 목적 산물인 카로티노이드 분석 결과에서도 모품종인 낙동벼에 비해 비타민A 강화벼에서 β-carotene은 10.6배 함량이 증가되고, zeaxanthin과 α-carotene는 생성되었음을 확인하였다. 이상의 분석기법을 통해 복수세대에서 비타민A 강화벼의 도입 유전자들이 안정적으로 유지되고 목적 단백질들이 안정적으로 발현되고 있음을 확인하였다.

Carotenoids are major secondary compounds in Citrus determining the color of fruit and nutritional values. Carotenoids are isoprenoic compounds, and function as color pigments in the flower and fruit to attract pollinators and seed-dispersing animal and chromophore for light harvest and photoprotectant during photosynthesis. In the aim of developing new cultivars with high value using molecular breeding technology, we had performed screening of flesh and peel specific genes by differentially expressed gene screening in Citrus unshiu fruits. From the screening, carotenoid isomerase (CRTISO)1, which converts pro-lycopene to all-trans-lycopene, was identified as peel-specifically expressed gene. In this study, the gene encoding the CRTISO1 was cloned, sequenced, and compared to the CRTISOs in other plant species. Comparison of the cds sequence to other plant species revealed 75% and 78% identity with CRTISO1 of Zea maize and CRTISO2 of Arabidopsis thaliana respectively. We also cloned CRTISO2 from C. unshiu which declines the expression while maturation (Kato et al., 2004), and the gene structure was analyzed. This is the first work reporting the full sequence and gene structure of CRTISOs in C. unshiu, and would give important information in understanding the carotenoid synthesis in the Citrus fruit.

Chinese cabbage (Brassica rapa ssp. pekinensis) is one of the most important vegetables and widely cultivated in Asia countries including Korea and China. Recently, whole genome sequence and full-length cDNA information of this species became available, which are encouraging genetic studies of this species to characterize agricultural important traits. Orange-colored (Or) cultivar of Chinese cabbage has inner leaves in orange, whereas other cultivars generally cultivated have yellow (Ye)- or white-colored inner leaves. In this study, we investigated phenotypes and carotenoid biosynthesis genes related to color variation in the Or cultivar. Firstly we compared the carotenoid content and composition between the Or and Ye cultivars by HPLC analysis. The inner leaves of Or cultivar contained approximately 9-fold high β-carotene content, whereas content of both lutein and violaxanthin was decreased to less than 30%, compared to Ye cultivar. Or cultivar was segregated with ratio of 3:1 in F2 population derived from crossing between Or and Ye inbred lines, indicating that Or phenotype is controlled by single recessive gene. To identify this gene, we investigated the expression of several genes involved in carotenoid biosynthesis by RT-PCR analysis. Among genes tested, two encoding putative carotenoid isomerase (CRTISO) and phytoene desaturase (PDS) were identified to show different expression between Or and Ye cultivars. Through further analysis of genomic DNA regions of these two genes, we could expect that several mutations such as InDel and base-substitution occurred and then affected expression of these genes in Or cultivar. In this presentation, I will introduce more detailed results for Or cultivars.

Suspended and sinking particles were collected during ODP Leg 119 to the Indian Ocean sector of the Antarctic Ocean. Field work was carried out at four sampling sites in Prydz Bay. Two of these sites were located in the Outer Bay, and two in the Inner Bay. At the four locations, a total of ten deployments of a sediment trap array were made. The concentrations of carotenoids both in suspended and sinking particulate matter in Prydz Bay were analyzed using HPLC. Fucoxanthin was the dominant carotenoid pigments both in suspended and sinking particles. The present study also indicates that 19'-hexanoyoxyfucoxanthin-containing prymesiophytes (Phaeocystis spp.) was abundant in the study area. The flux rates of carotenoids were generally highest at 50 m, and approximately double the flux rates at deeper horizons, however, at Inner Bay sites, the mean flux rates of carotenoids were greatest at 200 m, and 3 times greater than that of 50 m. Such anomalous high fluxes at 200 m imply that grazers were locally abundant between 100 m and 200 m at these sites close to land, and this hypothesis is supported by visual evidence of lots of fecal pellets in the 200 m trap. Integrates standing stocks versus sinking pigments data support that particulate material in Prydz Bay was not recycled rapidly.