Citrus Sinensis OSBECK Research Articles

Preliminary optimization of extraction process for Citrus sinensisOsbeck L. peel essential oil and quality evaluation

The present study performed optimization of the essential oil extraction from Citrus sinensis Osbeck L. peel. The conditions of grinding time of 1 min, extraction time of 150 min, and a water-to-material ratio of 1:3 (w/v) on frozen matter yielded 4.20 (%, g/100 g fresh matter) of extract in semi-pilot scaleby hydrodistillation method. C. sinensis peel EO was subjected to sensory evaluation, heavy metal contents, physical properties including density, pole rotation angles, and refractive index. D-limonene was identified in the synthesized peel extract with the highest content of 98.12 %. Thisprocess may have great potential as an alternative as it allows the recovery of C. sinensis by-products and production of high quality essential oils. ® 2024 Journal of Science and Technology - NTTU

Optimizing yields in sweet orange (Citrus sinensis osbeck) through phenology based nutrition

Optimizing yields in sweet orange (Citrus sinensis osbeck) through phenology based nutrition

Simultaneous qualitative and quantitative analysis of 10 bioactive flavonoids in Aurantii Fructus Immaturus (Zhishi) by ultrahigh-performance liquid chromatography and high-resolution tandem mass spectrometry combined with chemometric methods.

Aurantii Fructus Immaturus (Zhishi in Chinese) is the dried young fruit of Citrus aurantium L. (CA) and its cultivated varieties or Citrus sinensis Osbeck (CS). The content of flavonoids in different varieties of Zhishi may be significantly different. However, there is confusion about the botanical origin of Zhishi, and there is no reliable and systematic method to control Zhishi quality. We aimed to establish an ultrahigh-performance liquid chromatography method coupled with diode-array detection and high-resolution tandem mass spectrometry (UPLC-DAD-HRMS/MS) for the quantitative analysis of 10 flavonoids in Zhishi that could be used for quality control and botanical origin identification. A UPLC-DAD-HRMS/MS method was established for simultaneous identification and quantification of 10 flavonoids. Separation was performed on a Waters Acquity UPLC HSS T3 column (100 mm × 2.1mm, 1.8μm) with 0.1% formic acid and acetonitrile as mobile phase under gradient elution. MS was performed in positive and negative ionisation modes. The flavonoids in 41 batches were isolated and quantified. Zhishi of different botanical origins were identified by chemometrics. The results showed that the established method for the determination of 10 components was reliable and accurate. Chemometrics could be used to distinguish Zhishi of different botanical origins. There were significant differences in the contents of 10 flavonoids in samples of different botanical origins. The quantitative analysis method in this study can be used to accurately determine the content of 10 flavonoids and provide a chemical basis for quality control and botanical origin identification of Zhishi.

Micropropagation studies of sweet orange (citrus sinensisOsbeck) cv. Blood Red

This study was conducted to study in vitro technique, micropropagation of sweet orange (Citrus sinensis Os beck) cv. Blood Red. The impact of plant growth regulators on axillary bud proliferation in Blood Red was studied by using MS basal medium added with BAP (1.0, 3.0, 5.0 and 7.0 mg l−1) and NAA (0.1, 0.5, 1.0 and 1.5 mg l−1) for rooting IBA and NAA (0.1, 0.5, 1.0 and 2.0 mg l−1 each) either alone or in combinations. The maximum percent success, number of shoots, length of shoot and minimum number of days taken to bud break were obtained with BAP (3.0 mg l−1) and NAA (0.5 mg l−1). For rooting, lower concentrations of NAA and IBA up to 0.5 mg l−1 were found better in case of maximum percent success, number of roots, length of roots and minimum number of days taken to root initiation as compared to higher levels.

Transcriptional Profiling of Canker-Resistant Transgenic Sweet Orange (Citrus sinensisOsbeck) Constitutively Overexpressing a Spermidine Synthase Gene

Citrus canker disease caused by Xanthomonas citri subsp. citri (Xcc) is one of the most devastating diseases affecting the citrus industry worldwide. In our previous study, the canker-resistant transgenic sweet orange (Citrus sinensis Osbeck) plants were produced via constitutively overexpressing a spermidine synthase. To unravel the molecular mechanisms underlying Xcc resistance of the transgenic plants, in the present study global transcriptional profiling was compared between untransformed line (WT) and the transgenic line (TG9) by hybridizing with Affymetrix Citrus GeneChip. In total, 666 differentially expressed genes (DEGs) were identified, 448 upregulated, and 218 downregulated. The DEGs were classified into 33 categories after Gene ontology (GO) annotation, in which 68 genes are in response to stimulus and involved in immune system process, 12 genes are related to cell wall, and 13 genes belong to transcription factors. These genes and those related to starch and sucrose metabolism, glutathione metabolism, biosynthesis of phenylpropanoids, and plant hormones were hypothesized to play major roles in the canker resistance of TG9. Semiquantitative RT-PCR analysis showed that the transcript levels of several candidate genes in TG9 were significantly higher than in WT both before and after Xcc inoculation, indicating their potential association with canker disease.

Cloning, expression, purification and physical and kinetic characterization of the phosphoenolpyruvate carboxylase from orange ( Citrus sinensis osbeck var. Valencia) fruit juice sacs

Phosphoenolpyruvate (PEP) carboxylase (PEPCase) from orange fruit juice sacs has been cloned and heterogously expressed in high yield. The purified recombinant enzyme displays properties typical of plant PEPCase, including activation by sugar phosphates and inhibition by malate and citrate. Malate inhibition is weak in the physiological pH range, and the enzyme is also poorly affected by Glu and Asp, known inhibitors of C 3 plants PEPCases. However, it is strongly inhibited by citrate. Orange fruit PEPCase phosphorylation by mammalian protein kinase A decreased inhibition by malate. The enzyme presents an unusual high molecular mass in the absence of PEP, while in its presence it displays a more common tetrameric arrangement. The overall properties of the enzyme suggest that it is suited for organic acid synthesis and NADH reoxidation in the mature fruit. The present study provides the first analysis of a recombinant fruit PEPCase.

Analysis of Microsatellites in Citrus Unigenes

Simple sequence repeats (SSRs) were investigated in the unigene sequences from expressed sequence tags (EST) of sweet orange ( Citrus sinensis osbeck), trifoliate orange ( Poncirus trifoliata Raf.) and other citrus species and cultivars. A total of 37 802 citrus unigene sequences corresponding to 23.29 Mb were searched, resulting in the identification of 8 218 SSRs. Among them there were 4 913 (59.8%) mono-, 1 419 (17.3%) di-, 1 709 (20.8%) tri-, 114 (1.39%) tetra-, 23 (0.28%) penta- and 40 (0.49%) hexa-nucleotide SSRs. The estimated frequency of SSRs was approximately 1/2.8 kb, which could be extrapolated to 1 SSR-containing unigene in 4.6 unigenes. The maximum length of the SSR ranged from 40 to 105 bp depending on the repeating numbers of the motif in the SSR. The overall average length of SSRs was 20.9 bp. The frequencies of different SSR types (di-, tri-, tetra-, and penta-nucleotide repeats) were very similar between sweet orange and trifoliate orange. The mononucelotide repeats appeared to be the most abundant SSRs within sweet orange and trifoliate orange, followed by trimeric repeats. The adenine rich repeats such as A/T, AG, AT, AAG, AAAT, AAAG, AAAT, AAAAG, AAAAT etc. were predominant in each type of SSRs (mono-, di-, tri-, tetra-, and penta-), whereas the C/G, CG, CCG repeats were less abundant. Twenty-five primer pairs flanking EST-SSR loci were designed to detect the possible polymorphism of six citrus cultivars including sweet orange and trifoliate orange. The PCR result with all these 25 primer pairs revealed the existence of polymorphism within six citrus cultivars confirming that citrus EST database could be efficiently exploited for the development of gene-derived SSR markers.

Agrobacterium-mediated transformation and regeneration of Citrus spp. from suspension cells.

Agrobaclerium-mediated transformation of suspension cells of Citrus and regeneration of transgenic plants have been achieved using a binary vector and a cointegrative vector systems. Three embryo callus lines of 'Washington navel' orange (Citrus sinensis OSBECK), 'Ohta' ponkan and 'Kara' mandarin (C. reticulata BLANCO), and a pollen embyoid callus of 'Trovita' orange (C. sinelesis) were cocultlvated with two strains of A. tumefaciens, A415 or GV3010, for 3, 5 and 7 days. The former strain harbors a binary vector pTRA415 containing a neomycin phosphotransferase II (npt-II) gene. The later strain harbors a cointegrative vector pGV3850 : pMPK110-1lpt containing a hygromycin phosphotransferase (hpt) gene. Callus colonies were formed in calli of 'Washington navel' orange and 'Trovita' orange on selection media containing kanamycin or hygromycin. A relatively high frequency of transformation was observed with 3-day-cocultivation but a lower frequency was observed with 7-day-cocultivation. Embryoids were formed from kanamycin or hygromycin resistant calli and developed into plantlets. Transformation was confirmed on calli and a plantlet by Southern hybrydization.

Studies on physiologically active substances in citrus peel. Part IX. Structure and hypotensive effect of flavonoid glycosides in orange (Citrus sinensis OSBECK) peelings.

Studies on physiologically active substances in citrus peel. Part IX. Structure and hypotensive effect of flavonoid glycosides in orange (Citrus sinensis OSBECK) peelings.

Effect of carbohydrates and inhibitors of GA 3 biosynthesis on embryogenenic potential of salt tolerant and non-tolerant callus lines of orange ( Citrus sinensis osbeck)

Embryogenesis in callus lines of Citrus sinensis (orange) selected for tolerance to NaCl stress was compared to an unselected line. Addition of inhibitors of gibberellic acid (GA 3) synthesis, 2.6 × 10 −6M paclobutrazol or 6.2 × 10 −6M daminozide (butanedioic acid mono-(2-2-dimethylhydrazide) to the medium enhanced embryogenesis in the unselected line and two of the three salt tolerant lines, and also overcame embryo browning in the salt tolerant line R13. Sucrose as a carbon source was generally ineffective for embryogenesis. Two percent (w/v) glycerol was more effective than 2% (w/v) galactose, especially in older cultures. Formation of embryos was quicker in the non-selected line. Only a slight improvement in embryogenic potential was noted upon removal of salt tolerant lines for three passages from NaCl containing medium to basal medium without salt.

Effects of sucrose concentration, pH of media, and culture temperature on anther culture of citrus.

Effects of sucrose concentration, pH of media, and culture temperature on anther culture were studied using trifoliate orange (Poncifus trifoliata (L.) RAF.), sour orange (Citrus aurantium L.), and 'Trovita' orange (Citrus sinensis OSBECK). The most effective concentrations of sucrose in forming embryoids were 30g/l for trifoliate orange, 70g/l for sour orange, and 10g/l for 'Trovita' orange. As well, more calli were formed on the media containing more sucrose in all three species. The optimal pH of the media was pH 5 or 6 in forming embryoids or calli for three species. Embryoids effectively appeared at 28°C in trifoliate orange, at 24°C and 28°C in sour orange, and at 24°C in 'Trovita' orange. Calli formed well at 28°C and 24°C in trifoliate orange, at 28°C in sour orange, and at 24°C in 'Trovita' orange. These results suggest that the response of the anthers to the culture conditions differs among close species of citrus.

Acridone alkaloids. VII. Constituents of Citrus sinensis Osbeck var. brasiliensis Tanaka. Isolation and characterization of three new acridone alkaloids, and a new coumarin.

Fractionation and chromatography of the acetone extract of the root bark of Citrus sinensis OSBECK var. brasiliensis TANAKA (Rutaceae) afforded three new acridone alkaloids namely citrusinie-I, -II and citbrasine and a new coumarin, ethylsuberenol. The structures of citrusinine-I, -II, and citbrasine, and ethylasuberenol were characterized as 1a, 2a, 3, and 6, respectively. Citracridone-I (4), suberenol (7), suberosin (8), crenulatin (9), xanthoxyletin (10), xanthyletin (11), nordentatin (12), elemol (13), and p-hydroquinone were also isolated.

果樹の葉内水分不足に関する研究 (第3報)

Relationships between soil factors and leaf water potential were studied during summer for satsuma mandarin (Citrus unshiu MARC.) and maltese blood (Citrus sinensis OSBECK) trees.1. Leaf water potential measured before sunrise (ψψmax) was curvilinearly related to the soil matric potential in the main root zone. When soil matric potential dropped below pF 3.5 to 4.0, the slope of the curve began to increase rapidly.Even under the same atmospheric and soil moisture conditions, leaf water potentials during the day (ψψmin) were always higher for the trees grown in the granite soil than in the rhyolite soil. The difference described above may be due to the differences in soil properties such as hydraulic conductivity and porosity between the two soil types.2. Both soil matric potential and ψψmax for trees at the site with shallow soil profile continued to fall rapidly as days went on, whereas at the site with deeper soil profile the rate of decrease of the both values began to retard after ψψmax reached around -4.5 bars. Since no difference was found in the extent of root depth between the two sites, the difference should be attributed to the supplemental water flow from the subsoil.3. Root system of a tree was divided into two parts; one was held in moist soil (irrigated daily), the other in dry soil (nonirrigated). The moisture of nonirrigated soil decreased as days went on, the values of ψψmax, however, were remained constant according to the proportion of rootlets in moist soil.4. The submerging treatment was carried out by using trees grown in pots. Initially, ψψmin in this treatment was lower than that of control plants irrigated daily. After 5 to 7 days, however, ψψmin and leaf diffusion resistance in the submerged plants became abnormally higher than those of controls.