Part Of Cotyledon Research Articles

Biogenic iron oxide nanoparticles enhance callogenesis and regeneration pattern of recalcitrant Cicer arietinum L.

This study is the first report on the biosynthesized iron oxide nanoparticles (IONPs) which mediate in-vitro callus induction and shoot regeneration in economically important recalcitrant chickpea crop (Cicer arietinum L.). Here, we used leaf extract of Cymbopogon jwarancusa for the synthesis of IONPs in order to achieve a better biocompatibility. The bioactive compounds in C. jwarancusa leaf extract served as both reducing and capping agents in the fabrication process of IONPs. Field emission scanning electron microscopy (FE-SEM) revealed rods like surface morphology of IONPs with an average diameter of 50±0.2 nm. Energy-dispersive X-ray spectroscopy (EDS) depicted formation of pure IONPs with 69.84% Fe and 30.16% O2. X-ray diffractometry (XRD) and attenuated total reflectance-fourier transform infrared (ATR-FTIR) validate the crystalline structure, chemical analysis detect the presence of various biomolecular fingerprints in the as synthesized IONPs. UV-visible absorption spectroscopy depicts activity of IONPs under visible light. Thermo-gravimetric analysis (TGA) displayed thermal loss of organic capping around 500°C and confirmed their stabilization. The biosynthesized IONPs revealed promising results in callus induction, shoot regeneration and root induction of chickpea plants. Both chickpea varieties Punjab-Noor 09 and Bittle-98 explants, Embryo axes (EA) and Embryo axes plus adjacent part of cotyledon (EXC) demonstrated dose-dependent response. Among all explants, EXC of Punjab-Noor variety showed the highest callogenesis (96%) and shoot regeneration frequency (88%), while root induction frequency was also increased to 83%. Iron content was quantified in regenerated chickpea varieties through inductively coupled plasma-optical emission spectrometry. The quantity of iron is significantly increased in Punjab-Noor regenerated plants (4.88 mg/g) as compare to control treated plants (2.42 mg/g). We found that IONPs enhance chickpea growth pattern and keep regenerated plantlets infection free by providing an optimum environment for rapid growth and development. Thus, IONPs synthesized through green process can be utilized in tissue culture studies in other important recalcitrant legumes crops.

Temporal and Spatial Expression Analysis of Shoot-Regeneration Regulatory Genes during the Adventitious Shoot Formation in Hypocotyl and Cotyledon Explants of Tomato (CV. Micro-Tom).

Enhancing the competence for plant regeneration in tissue culture studies is an important issue not only for efficient genetic transformation of commercial crops but also for the reproducibility of scientific reports. In this study, we investigated optimization of several tissue culture conditions including plant growth regulators, types and ages of explants, culture densities, and plant position in order to improve the competence of adventitious shoot formation of the tomato (Solanum lycopersicum cv. Micro-Tom). In addition, we examined the differential expression of D-type cyclin (CYCD3-1) and several shoot regeneration regulatory genes from hypocotyl and cotyledon explants of tomato during shoot organogenesis. A treatment of 1 mg L−1 Zeatin and 0.1 mg L−1 Indole-3-acetic acid (IAA) in Murashige and Skoog (MS) medium containing 3% sucrose was optimal for adventitious shoot formation from hypocotyl and cotyledon explants. The younger explants exhibited more shoot formation regardless of explant types. Additionally, those closest to the shoot apical meristem produced more shoots compared to the other regions in the hypocotyl and the cotyledon explants. Gene expression of CYCD3-1, SHOOT MERISTEMLESS (STM), and cytokinin dependent WUSCHEL (WUS) was significantly higher in younger explants than in older ones. Furthermore, an increase in CYCD3-1, STM, and WUS expression was evident at the distal part of hypocotyls and the proximal part of cotyledons compared to other regions. These differential gene expression profiles exhibited good agreement with the results of shoot formation obtained from diverse explants of tomato. These results suggest that temporal and spatial gene expression of shoot regeneration regulatory genes plays an important role in enhancing the competence and the reproducibility of adventitious shoot formation from tomato explants.

Isoflavone profiles and antioxidant properties in different parts of soybean sprout.

Sprouting is one of the processing methods to enhance bioavailability of some nutrients of agricultural products. Soybeans were germinated for 6 days and soybean (SB), soybean sprout (SBS), cotyledon of soybean sprout (CSS), and hypocotyl and root of soybean sprout (HRSS) were collected. Isoflavone profiles and in vitro antioxidant activities of 80% ethanol extracts of samples were analyzed. In addition, oxidative stability of the extract was evaluated in corn oils heated at 100 °C.Thehypocotyl and root part of soybean sprout contained significantly (P<0.05) higher content of isoflavone aglycones than the cotyledon part of soybean sprout. Diphenyl-1-picrylhydrazyl radial and 2,2'-azinobis(3-ethylbenzothiazoline-6-sul-fonic acid cation radical scavenging activities of SBS were 1.6 and 1.3 times higher than those of SB, respectively (P <0.05), whereas those of CSS were 1.1 and 1.8 times higher than those of HRSS, respectively (P<0.05). CSS had higher ORAC and FRAP values than others. High correlations between contents of total flavonoids and antioxidant activities were observed in all in vitro antioxidant assays. SBS, CSS, and HRSS had significantly higher oxidative stability than SB in 100 °C heated corn oil after 9 hr treatment (P <0.05). Generally, 80% ethanol extract of CSS had high in vitro antioxidant properties whereas HRSS had high aglycone forms of isoflavones. Isoflavone aglycones may not be major ingredients providing antioxidant activities of soybean sprout. PRACTICAL APPLICATION: Soybeans are important crops for the preparation of tofu, soymilk, and protein sources. Soybean sprout prepared by germinating soybean and growing for several days is one of important food materials used as daily diet in some countries. In this study, antioxidant activities of different parts of soybean sprouts were evaluated using in vitro methods and bulk oil model systems. In addition, isoflavone profiles of soybean sprouts were determined. Enhanced antioxidant activities of soybean sprouts may help consumers choose healthier foods and extend business areas of the soybean food industry.

In vitro regeneration through organogenesis in Egyptian chickpea

Genetic engineering for improvement of the recalcitrant crop chickpea (Cicer arietinum L.) was largely restricted by the lack of an efficient regeneration system. In vitro regeneration in two Egyptian chickpea varieties, Giza 531 and Giza 4 was achieved by direct organogenesis. A variety of embryo explants and different types and concentrations of growth regulators were investigated for maximum efficiency of shoot and root regeneration. Embryo axes with the adjacent part of cotyledon proved to be the most promising type of explant for shooting and rooting responses. 6-Benzylaminopurine (BAP) and indole-3-butyric acid (IBA) were found to induce the highest percentages of shoot initiation and root formation, respectively. Although the Giza 531 variety produced a better response than the Giza 4 for shoot formation, it displayed lower performance for root induction. It would be rewarding if this optimized regeneration protocol paved the way toward the genetic improvement of the Egyptian chickpea.

Roles of oxides of nitrogen on quality enhancement of soybean sprout during hydroponic production using plasma discharged water recycling technology

This study was performed to assess the effect of plasma-discharged water recycling technology as irrigation water on soybean sprout production. Two different types of irrigation water were used individually for cultivation, including plasma discharged water as a source of oxides of nitrogen and tap water, irrigation water was recycled for every 30 minutes. Plasma discharged irrigation water reduced overall 4.3 log CFU/ml aerobic microbe and 7.0 log CFU/ml of artificially inoculated S. Typhimurium within 5 minutes and 2 minutes, respectively, therefore sprout production occurs in a hygienic environment. Using of plasma-discharged water for cultivation, increases the amount of ascorbate, asparagine, and γ-aminobutyric acid (GABA) significantly (p < 0.05), in the part of cotyledon and hypocotyl of soybean sprout during 1 to 4 days of farming. A NO scavenger, 2-(4-carboxy-phenyl)-4,4,5,5-tetramethylimidazoline-1-oxy-3-oxide (cPTIO), was added in irrigation water to elucidate the roles of the oxides of nitrogen such as NO3−, NO2− generated in plasma discharged water. It was observed that all three nutrients decreased in the cotyledon part, whereas ascorbate and GABA contents increased in the hypocotyl and radicle part of bean sprout for the same duration of farming. The addition of NO scavenger in the irrigation water also reduced growth and overall yield of the soybean sprouts. A recycling water system with plasma-discharged water helped to reduce the amount of water consumption and allowed soybean sprouts growth in a hygienic environment during the hydroponic production.

Adventitious regeneration in styrian oil pumpkin in relation to the endoreduplication pattern and induced tetraploidy on fusaric acid-supplemented media

We attempted to improve the adventitious regeneration protocol in styrian oil pumpkin for subsequent use in somaclonal variation studies. The endoreduplication status of tissues might play an important role, so endopolyploidy was analyzed by flow cytometry in different organs and major differences were revealed. Endoreduplication was minimal in leaves but in other organs—particularly in the hypocotyl and epicotyl—levels up to 64C were found. The basal part of cotyledons has been reported to be the most responsive for adventitious regeneration and, using image cytometry, we identified it as the least endoreduplicated section, with a cycle value of 1.29 compared to 1.78 and 1.80 in the central and distal parts. Basal cotyledonary explants were subjected to various media combinations, revealing Murashige and Skoog (Physiol Plant 15:473–497, 1962, doi:10.1111/j.1399-3054.1962.tb08052.x) medium supplemented with 1 mg L−1N6-benzylaminopurine (BA), 0.25 mg L−1p-aminobenzoic acid (PABA) and 5 mg L−1 fusaric acid (FA) to be optimal (73.3 % regenerated explants with 1.8 shoots per regenerating explants). FA was added to media as a possible selective agent for somaclonal variation inducing increased tolerance to Fusarium but was surprisingly found to stimulate regeneration at a low concentration (5 mg L−1) and to induce genome doubling at medium concentrations (10 and 20 mg L−1).

Two ways of plant regeneration from immature cotyledons of pea

Two different systems of plant regeneration via organogenesis and embryogenesis from immature cotyledons of pea (&lt;i&gt;Pisum sativum&lt;/i&gt; L.) were developed. The first system was direct multiple shoot regeneration from the proximal to the embryo axis, injured part of cotyledon. The ability of six Polish cultivars to shoot formation was very high. The percent of regenerating cotyledons was from 73 to 86 and mean number of shoots from 3.5 to 9.9 after seven weeks of culture. This multiplification could be prolonged for next several months. The second system was somatic embryogenesis, initiating from the same part of cotyledon simultaneously with slowly proliferating callus. Only three out of six cultivars formed embryoids. The differences of ability to embryo formation ranged between 43% of responding explants for Heiga cultivar to only 6% for Cud Ameryki. The mean number of embryoids was from 4.2 to 2.3.

In vitro Plant Regeneration System for Two Middle East Cultivars of Chickpea (Cicer arietinum L.)

This study surveyed a rapid, efficient and reproducible protocol for in vitro shoot regeneration and rooting of two Middle East chickpea cultivars (Balila and Wady). Organogenic ability of various explants (decapitated embryo axes, fragments of embryo axes, fragments of embryo axes with basal part of cotyledons, entire and half of cotyledons) was tested on MS media containing different concentrations and nature of cytokinins (BAP and zeatin) or compound with cytokinin-like activity (TDZ). The highest level of shoot regeneration was achieved in Balila genotype using embryo axes fragments including the basal part of cotyledon on medium with 0.25 mg/L BAP. Higher BAP concentrations than 0.25 mg/L for Balila and 0.5 mg/L for Wady caused a decrease of shoot regeneration. On MS media supplemented with zeatin (1, 3 and 5 mg/L) and thidiazuron (1, 3 and 5 mg/L), the highest level of shoot differentiation was obtained from calli derived from fragments of embryo axes. Organogenic ability of calli from Wady genotype was enhanced by increasing zeatin and TDZ concentrations, up to 5 mg/L. Conversely, a progressive decrease of shoot frequency for Balila and root development for both genotypes, were observed with increasing concentrations of BAP, zeatin and TDZ.

Induction of shoot buds, multiplication and plantlet formation in seedling explants of bell pepper (Capsicum annuum L.) cv. Bryza in vitro

In vitro shoot bud induction and multiplication as well as plantlets formation from different parts of 21-d old seedlings (shoot tip, cotyledonary node, distal part of cotyledon, acropetal section of hypocotyl) of &lt;em&gt;Capsicum annuum&lt;/em&gt; L., cv. Bryza were compared. During 4 weeks of primary explant culture on initiation media, first shoot bud primordia appeared; they reminded leaf primordia and subsequently some of them underwent enlargement, some developed into leaves and leaf-like structures (mainly on cotyledon explants). The highest number of shoot bud primordia was noted on cotyledonary node explants, but they were smaller than those on the remaining types of the explants. The best response of shoot regeneration showed cotyledon explants on which most of shoot buds were formed in each from four treated passages. From shoot buds on elongation media after 4 weeks of culture rooted rosettes of leaves were achieved, and the extension of the culture time to eight weeks without subculture caused that the rosettes developed into plantlets. Throughout four successive passages plantlets were obtained from cotyledon and shoot tip explants.

Characterization of human placental glycosaminoglycans and regional binding to VAR2CSA in malaria infected erythrocytes

Placental malaria is a serious problem in sub-Saharan Africa. Young women are particular susceptible to contracting this form of malaria during their first or second pregnancy despite previously acquired immunity from past infections. Placental malaria is caused by Plasmodium falciparum parasites expressing VAR2CSA on the erythrocyte surface. This protein adheres to a low-sulfated chondroitin sulfate-A found in placental tissue causing great harm to both mother and developing fetus. In rare cases, the localization of infected erythrocytes to the placenta can even result in the vertical transmission of malaria. In an effort to better understand this infection, chondroitin sulfate was isolated from the cotyledon part of the placenta, which should be accessible for parasite adhesion, as well as two non-accessible parts of the placenta to serve as controls. The placental chondroitin sulfate structures and their VAR2CSA binding were characterized. All portions of human placenta contained sufficient amounts of the appropriate low-sulfated chondroitin sulfate-A to display high-affinity binding to a recombinant truncated VAR2CSA construct, as determined using surface plasmon resonance. The cotyledon is the only placental tissue accessible to parasites in the bloodstream, suggesting it is the primary receptor for parasite infected red blood cells.

동부나물의 부위별 생육, 폴리페놀 및 항산화성 차이

새싹재배기에서 7일간 재배된 동부 새싹나물의 부위별 생육, 폴리페놀 함량, 플라보노이드 함량, 항산화성 및 항산화효소 활성 차이를 검토하였다. Folin-Denis방법에 따른 총 페놀 함량은 동부나물 자엽의 메탄올 추출물(48.8 mg <TEX>$kg^{-1}$</TEX>)이 가장 높았으며, 그 다음이 뿌리(30.8 mg <TEX>$kg^{-1}$</TEX>), 하배축(22.2 mg <TEX>$kg^{-1}$</TEX>) 순으로 나타났다(p < 0.05). 한편, 총 플라보노이드 함량은 총 페놀 함량과 같은 경향을 보였으나 더 낮은 함량이 검출되었다. DPPH 라디컬 소거능은 추출물 농도가 증가할수록 높은 활성을 보였으며 동부나물 자엽의 추출물(82.5%)에서 가장 높았으며, 그 다음이 뿌리(52.6%), 하배축(35.0%) 순으로 나타났다(p < 0.05). 항산화효소 활성은 APX와 CAT 활성은 하배축에서 가장 높았고 POX와 SOD 활성은 뿌리가 가장 높은 활성을 보였다. 이런 경향은 재배 후 5일과 7일째 동부나물에서 같게 나타났다. 따라서 상관분석에 따르면 총 페놀 함량이 총 플라보노이드 함량보다 항산화성과 항산화효소 활성에 더 높은 관련성이 있는 것으로 나타났고 그 생리활성물질 함량과 그 활성 정도는 부위별로 다르게 나타났다. An experiment was conducted to determine the content of phenolics and flavonoids, antioxidant activity and antioxidant enzyme status for the extracts from 5 and 7-day old sprouts (DOS) of cowpea (Vigna unguiculata L. Walp). Total phenolics [mg ferulic acid equivalents (FAE) <TEX>$kg^{-1}$</TEX> DW] content was highest in cotyledon extracts (48.8 mg <TEX>$kg^{-1}$</TEX>), followed by roots (30.8 mg <TEX>$kg^{-1}$</TEX>) and hypocotyl (22.2 mg <TEX>$kg^{-1}$</TEX>) extracts (p < 0.05) from 5 DOS. The result of total flavonoid level [mg rutin equivalents <TEX>$kg^{-1}$</TEX> DW] had same tendency to the results of total phenolics, showing lower amount ranges. The antioxidant activity of the methanol extracts from all the plant dose-dependently increased. DPPH (1,1-diphenyl-2-picryl hydrazyl radical) free radical scavenging activity was higher in cotyledon extracts (82.5%) than in root (52.6%) or hypocotyl parts (35.0%) from 5 DOS. Among antioxidant enzymes, APX and CAT activities were highest in cotyledon part and POX and SOD activities in root part of 5 and 7 DOS. The results showed that total phenolics content (<TEX>$r^2$</TEX> = 0.1516~0.9911) were more highly correlated with antioxidant activity than total flavonoids level (<TEX>$r^2$</TEX> = 0.0113~0.9442), and that the level and activity of physiological-active substances were different depending on plant part of the sprout.

Expression of the BBM gene during somatic embryogenesis of Arabidopsis thaliana

The relationship between somatic embryogenesis (SE) and the expression of the BABY BOOM (BBM) gene was studied in cultured immature zygotic embryos (IZEs) using a transgenic line of Arabidopsis thaliana containing a BBMPro::GUS construct. Results showed spatio-temporal differences in BBM expression in explants during culture. BBM promoter activity was observed in freshly isolated IZEs except distal parts of cotyledons. At the beginning of culture, considerable increase of GUS staining intensity was observed in all parts of explants, which maintained at high level over next few days and coincide with cell divisions. Gradual decrease of GUS distribution in explants was observed at about the 5th day of culture. BBM promoter activity became largely restricted to dividing cells, then to developing somatic embryos, shoot-like structures and callus. In parts of explants not involved in morphogenesis BBM promoter activity was absent or hardly seen. Thus the in vitro expression of BBM coincides with cell proliferation and morphogenesis.

Expression of the BBM gene during somatic embryogenesis of Arabidopsis thaliana

The relationship between somatic embryogenesis (SE) and the expression of the BABY BOOM (BBM) gene was studied in cultured immature zygotic embryos (IZEs) using a transgenic line of Arabidopsis thaliana containing a BBMPro::GUS construct. Results showed spatio-temporal differences in BBM expression in explants during culture. BBM promoter activity was observed in freshly isolated IZEs except distal parts of cotyledons. At the beginning of culture, considerable increase of GUS staining intensity was observed in all parts of explants, which maintained at high level over next few days and coincide with cell divisions. Gradual decrease of GUS distribution in explants was observed at about the 5th day of culture. BBM promoter activity became largely restricted to dividing cells, then to developing somatic embryos, shoot-like structures and callus. In parts of explants not involved in morphogenesis BBM promoter activity was absent or hardly seen. Thus the in vitro expression of BBM coincides with cell proliferation and morphogenesis.

Factors Affecting Regeneration of Tomato Cotyledons

In this research, we employed the tomato cultivar named Zhongshu Si Hao as research material to study the influences on the regeneration of adventitious buds under the different conditions with hormone combinations, various explants, different parts of cotyledons and cotyledon inoculatingmethods; we also screened the best method for seed surface disinfection and the most appropriate medium for the rooting of adventitious buds. The results showed that the best procedures for surface disinfection of tomato seeds would be as follows: firstly, immersing the seeds in 70% ethanol for 30 second, then rinsing them in 20% NaClO with tween-20 for 20 min. The most ideal explant for inducing buds is the middle part of the cotyledon, followed by the stem and the leaf. the best medium formula for cotyledon regeneration is MS+ZT 2 mg/L+IAA 0.1 mg/L, and placing the cotyledon upside down is better to induce the adventitious buds than that placing upside. The best medium for rooting of adventitious buds is 1/2MS+IAA 0.3 mg/L. This research would lay a foundation establish transformation system of tomato in our future research.

콩나물의 지방산 및 식이섬유 함량 분석

Soy sprout is a traditional food in Korea. It has been a source of protein, lipid and vitamin etc. as cooked and fresh vegetable supplying throughout the year. Soy sprouts of five soybean cultivars were analyzed fatty acid composition and dietary fiber contents. The compositional analysis confirmed that soy sprout contained five major fatty acids( 16:0 palmitic, 18:0 stearic, 18:1 oleic, 18:2 linoleic and 18:3 linolenic). Fatty acid levels in soy sprout were not significant statistically between fresh and boiled soy sprout, the result support a conclusion that boiling treatment could not affect the fatty acid composition in soy sprouts. Fatty acid levels in whole and cotyledon part of soy sprout were not changed during sprout production, whereas the content in hypocotyl increased by approximately 13.2% of total unsaturated fatty acid compared to whole soy sprout. In case of dietary fiber, boiled soy sprouts showed higher contents than in fresh soy sprouts. These changes were associated with an increase in the mean level of insoluble dietary fiber expecially. Fatty acid levels and dietary fiber contents varied with soybean cultivars suggesting chemical composition of soybean seed was important to soy sprout characters.

Stable integration and expression of a plant defensin in tomato confers resistance to fusarium wilt

Plant defensins are small cysteine-rich peptides which belong to a group of pathogenasis related defense mechanism proteins. The proteins inhibit the growth of a broad range of microbes and are highly stable under extreme environmental stresses. Tomato cultivation is affected by fungal disease such as Fusarium wilt. In order to overcome fungal damages, transgenic tomato plants expressing the Medicago sativa defensin gene MsDef1 under the control of the CaMV 35S promoter were developed. The Fusarium-susceptible tomato (Lycobersicum esculentum Mill) cultivar CastleRock was used for transformation to acquire fungal resistance. Hypocotyl with a part of cotyledon (hypocotyledonary) for young tomato seedlings were used as an explant material and transformation was performed using the biolistic delivery system. Bombarded shoots were selected on regeneration medium supplemented with hygromycin and suitable concentrations of BA, zeatin ripozide and AgNO3. Putative transgenic plantlets of T0 were confirmed by PCR analysis using primers specific for the transgene and the transformation frequency obtained was 52.3%. Transformation and transcription of transgenes were confirmed in T1 by PCR, Southern hybridizations, and reverse-transcription PCR (RT-PCR). The copy numbers of integrated transgene into tomato genome ranged between 1-3 copies. Greenhouse bioassay was performed on the transgenic T1 and T2 young seedlings and non-transgenic controls by challenging with a vigorous isolate of the fungal pathogen Fusarium oxysporum f. sp. Lycopersici. The level of fungal infectivity was determined using RT-PCR with tomatinase specific primers. Transgenic lines were more resistant to infection by fusarium than the control plants. These results indicated that overexpressing defensins in transgenic plants confer resistance to fungal pathogens.

Arabidopsis EMBRYOMAKER encoding an AP2 domain transcription factor plays a key role in developmental change from vegetative to embryonic phase

Although several types of plant cells retain the competence to enter into embryonic development without fertilization, the molecular mechanism(s) underlying ectopic embryogenesis is largely unknown. To gain insight into this mechanism, in a previous study we identified 136 ESTs specifically expressed in microspore embryogenesis of Brassica napus. Here, we describe the characterization of the Arabidopsis EMBRYOMAKER (EMK) gene, which is homologous to one of the identified Brassica ESTs (BnGemb-58) and encodes an AP2 domain transcription factor. The AtEMK was expressed in developing and mature embryos, but its rapid disappearance occurred during germination. After germination, the expression of AtEMK was found in the root apical meristem and the distal parts of cotyledons. Although a mutant lacking AtEMK exhibited no distinctive defects in the embryo, ectopic expression of AtEMK induced embryo-like structures from cotyledons. The embryo-like structures contained high concentration of lipids, expressed several embryo-specific genes, and could convert into independent plants, indicating that the structures are somatic embryos. In vitro culture, AtEMK enhanced the efficiency of somatic embryogenesis. Furthermore, ectopic expression of AtEMK caused the formation of trichomes on cotyledons, dedifferentiated several tissues into calli, and retarded root development, demonstrating that AtEMK is harmful for the normal development of plants after germination. From these results, we conclude that the AtEMK is a key player to maintain embryonic identity, and the rapid disappearance of AtEMK expression during germination is essential for the developmental transition between the embryonic and vegetative phases in plants.

SOMACLONAL VARIATION AMONG SOMATIC-EMBRYO DERIVED PLANTS OF OLEA EUROPAEA L “CV. KRONEIKI”

Frequency of somaclonal variation in embryogenic calli and plants of Olea europaea L cv.Kroneiki derived from somatic embryogenesis were examined by randomly amplified polymorphic DNA (RAPD) analysis. Radicle and cotyledon (proximal part) of mature zygotic embryo were cultured in OMc medium supplemented with IBA (5 mgL-1) and 2-ip (0.5 mgL-1). Somatic embryogenesis was induced in calli of radicles (55%) and proximal part of cotyledons (8%) on OM medium without any hormones. After culturing the embryogenic calli on OM medium in the presence of BAP (2 mgL-1), regenerated plants were obtained. DNA sample from the leave of the mother plants, seedlings (germinated from zygotic embryo), plantlets (regenerated from somatic embryo), embryogenic calli, non-embryogenic calli and zygotic embryos were subjected to RAPD analysis. 20 arbitrary decamer primers produced polymorphic amplification products. These primers gave a total of 315 reproducible fragments from which 221 (71%) were useful as polymorphic bands with an average of twelve RAPD marks obtained for each primer. A dandogram, based on the unweighted pair group mean average (UPGMA) method of cluster analysis, were constructed using a similarity matrix derived from the RAPD amplification generated by all primers. The estimation of genetic similarity coefficient based on RAPD band sharing data indicated that regenerated plants were less than 75% similar to mother plants.

콩나물의 재배기간과 부위별 TDF에 결합된 주요 무기질의 변화와 추정 이용율

This study was conducted to determine an estimated value and change in TDF (total dietary fiber) binding major minerals (Ca, P, K Mg) of soybean sprouts depending on cultivation periods of 1, 3 or 5 days, and parts of cotyledon and hypocotyl. The compositions of TDF binding major minerals in soybean sprouts were evaluated by the enzymatic-gravimetric method developed by Prosky and adopted by AOAC. The content of TDF binding P ranged between 915.28 mg and 1037.82 mg per 100 g cotyledon. Average contents of Ca, K, and Mg in cotyledon ranged between 541.67 mg and 634.34 mg, 180.91 mg and 253.98 mg, 231.90 mg and 301.01 mg, respectively, on dry matter basis. The average contents of TDF binding Ca in hypocotyl per 100 g were between 454.36 mg and 540.33 mg, and other major minerals contents were between 149.24 mg and 186.21 mg for P, 164.17 mg and 182.78 mg for K, and 152.53 mg and 161.22 mg for Mg, respectively. The proportional changes of Ca, an estimated value in cotyledon, ranged between 5.58% and 30.98%, 47.18% and 59.10% for P, 95.24% and 95.50% for K, and 58.45% and 68.64% for Mg, respectively, based on dry matter. An estimated value of Ca, P, K, and Mg in hypocotyl ranged between 6.71% and 13.52%, 91.03% and 91.76%, 96.91% and 98.92%, and 61.03% and 66.37%, respectively, on dry matter basis.

In vitro Regeneration of Garden Balsam, Impatiens balsamina Using Cotyledons Derived from Seedlings

Garden balsam, Impatiens balsamina, is one of an important ornamental plant in Malaysia. An effort to develop a reliable tissue culture system for this Impatiens species was carried out for future genetic manipulation of this plant for useful traits. Parts of cotyledons and hypocotyl derived from different age of seedlings, from surface sterilized seeds, were subjected to shoot induction on MS media containing BAP, TDZ or combination of BAP and NAA in different ratios. Roots were developed from the above shoots on full- or halfstrength MS media containing IAA, IBA or NAA of different concentrations. Finally fully-developed plantlets were produced on hormone free MS media or full- or half-strength MS media containing specific hormones. It was observed that the most efficient shoot induction (93%) was obtained from proximal part of cotyledon derived from 7 days old seedlings on MS media supplemented with 1mg/L BAP. Roots were most efficiently produced (92%), in term of percentage and morphology, on half-strength MS media supplemented with 0.1 mg/L IAA. Further regeneration of plantlets, approximately 8-10 cm in height, was achieved after 3 weeks on hormone-free MS media. Finally, full regeneration system of Impatiens balsamina was developed, using MS media supplemented with 1 mg/L BAP for shoot and 0.1 mg/L IAA for roots, within 8 weeks in culture.