Monoterpenoids from the fruit of Gardenia jasminoides Ellis (Rubiaceae)

The present phytochemical investigations of Gardenia jasminoides Ellis resulted in the isolation of ten iridoids (1–10) and ten pyronane monoterpenoids (11–20). Among them, compounds 11 and 18 were obtained from this species for the first time. The chemotaxonomic importance of these compounds was also summarized.     

大孔吸附树脂对栀子中环烯醚萜苷类成分的富集行为

采用大孔吸附树脂对栀子中环烯醚萜苷类成分的富集行为进行研究,结果表明,D101大孔吸附树脂柱层析适合分离纯化栀子中环烯醚萜苷类成分,采用80%乙醇洗脱,紫外-可见分光光度法进行定量测定。使用该方法富集后,80%乙醇洗脱液干燥后总固体物中环烯醚萜苷类成分含量可达到70%以上。     

Iridoids from the flowers of Gardenia jasminoides Ellis and their chemotaxonomic significance

Phytochemical investigation of the flowers of Gardenia jasminoides Ellis (Rubiaceae Gardenia) resulted in the isolation and identification of four iridoid aglycones (1–4) and eleven iridoid glycosides (5–15). This is the first report of the occurrence of these compounds in the genus Gardenia: garjasmine (1), dunnisin (2), α-gardiol (3), β-gardiol (4), diffusoside A (6), diffusoside B (7), genameside C (13), and deacetylasperulosidic acid (14). The chemotaxonomic significance and biosynthetic pathways of these iridoid aglycones (1–4) and iridoid glycosides (5–15) are summarized. Iridoid aglycones and iridoid glycosides are considered as important chemotaxonomic markers in Rubiaceae and this work indicates that the iridoid aglycones (1–4) have a limited distribution in the Rubiaceae.     

高效液相色谱法测定山栀子中栀子甙的含量

采用高效液相色谱法测定中药山栀子（Cardenia jasminoides Ellis）中保肝利胆的主要成分栀子甙（geniposide）的含量。分析结果表明,本方法重现性好,平均加样回收率为100．90％,RSD为2．00％。用此方法测定未知样品山栀子中栀子甙的平均含量为3．463g/100g。     

Effects of Light and Sucrose Levels on the Anatomy, Ultrastructure, and Photosynthesis of Gardenia jasminoides Ellis Leaflets Cultured in vitro

This article reports the effect of growing conditions on the anatomical and ultrastructural changes associated with the development of photoautotrophy in Gardenia jasminoides Ellis plantlets during shoot multiplication in vitro. Two photosynthetic photon flux densities (PPFD) (50 and 100 μmol m-2 s-1 PPFD: L50 and L100, respectively) and two sucrose concentrations in the culture medium (5 and 30 g L-1: S5 and S30, respectively) were assayed. An increase in PPFD stimulated the development of photosynthetic tissues and led to higher photosynthesis and dark respiration regardless of the sucrose level assayed. However, the effect of sucrose in the medium depended on the PPFD. For the high-PPFD treatment, a low sucrose concentration in the medium stimulated the development of photosynthetic tissues, whereas the opposite effect of sucrose was observed at low PPFD. This study demonstrated that an increase in light intensity to moderate values such as L100 has a beneficial effect on the development of structural changes associated with photoautotrophy. Such an effect is stimulated by low sucrose (S5) levels in the medium. These modifications of usual growing conditions (such as L50 combined with S30) for micropropagation may prove to be useful in mass propagation of gardenia. However, the use of a low sucrose level in addition to conventional growing PPFD may be counterproductive.     

Rapid in vitro multiplication and ex vitro rooting of Malus zumi (Matsumura) Rehd

Micropropagation system of Malus zumi was optimized by studying the influence of plant growth regulators and culture conditions. The axillary buds were used for mutiplication of in vitro shoot culture on agar Murashige and Skoog (1962) (MS) medium with combination of 1 mg l⁻¹ BAP, 0.5 mg l⁻¹ NAA or 0.5 mg l⁻¹ IAA or 0.5 mg l⁻¹ IBA under 16 h photoperiod. The shoot growth in culture was not significantly affected within a broad range (5.0–7.0) of initial medium pH. The highest shoot (13) was obtained on medium containing 1.0 mg l⁻¹ BAP and 0.5 mg l⁻¹ IAA. Well-developed shoots, 35–50 mm in length, were successfully rooted ex vitro at 86.3% by a 2-h-treatment with aqueous solution containing MS salts and 100 mg l⁻¹ IBA prior to their planting in growing substrate composed of soil and vermiculite (1:1 v/v). The survival rate of transplantation reached 88.0% when transferred to field condition.     

Activity and isoforms of peroxidases, lignin and anatomy, during adventitious rooting in cuttings of Ebenus cretica L

Adventitious rooting of Ebenus cretica cuttings was studied in order to examine a) the rooting ability of different genotypes in relation to electrophoretic patterns of peroxidases. b) the activity and electrophoretic patterns of soluble and wall ionically bound peroxidases, the lignin content and anatomical changes in the control and IBA treated cuttings of and genotypes in the course of adventitious root formation. In addition, a fraction of soluble cationic peroxidases was separated by gel filtration chromatography from the total soluble peroxidases of a genotype. No rooting occurred in cuttings without IBA-treatment. In both genotypes, electrophoretic patterns of soluble anionic peroxidases revealed two common peroxidase isoforms, while a fast-migrating anionic peroxidase isoform (A3) appeared only in genotypes. Both genotypes showed similar patterns of soluble, as well as wall ionically bound cationic peroxidase isoforms. The number of isoforms was unchanged during the rooting process (induction, initiation and expression phase) but an increase in peroxidase activity (initiation phase) followed by decrease has been found in IBA-treated cuttings. During initiation phase the lignin content was almost similar to that on day 0 in genotype while it was reduced at by about 50% in genotype at the respective time. Microscopic observations revealed anatomical differences between genotypes. According to this study, the and genotypes display differences in anatomy, lignin content, activity of soluble peroxidases and the electrophoretic patterns of soluble anionic peroxidase isoforms. The A3-anionic peroxidase isoform could be used as biochemical marker to distinguish and genotypes of E. cretica and seems to be correlated to lignin synthesis in rooting process.     

The gibberellin synthesis inhibitors,ancymidol and flurprimidol,promote in vitro rooting of white pine microshoots

Summary Ancymidol and flurprimidol were tested for their ability to induce in vitro rooting on axillary proliferated shoots of white pine (Pinus strobus L.). Shoots were treated for 30 days (pulse) with growth regulators, then subcultured to 0.5X medium for conifer morphogenesis without growth regulators. A pulse treatment containing 5 M ancymidol and 0.54 M naphthaleneacetic acid resulted in 43% rooted shoots, whereas a pulse treatment with 0.54 M naphthaleneacetic acid alone resulted in 7% root formation. Flurprimidol also stimulated rooting of white pine shoots, but was less effective than ancymidol. No detrimental effects on shoot growth were observed with the gibberellin synthesis inhibitors at the 5 M concentration used. Some rooted shoots were successfully acclimatized to the greenhouse.Abbreviations ancymidol -cyclopropyl-(4-methoxyphenyl)-5-pyrimidinemethanol - flurprimidol -(1-methylethyl--[4-trifluromethoxy)phenyl]-5-pyridinemethanol - GA gibberellin - MCM medium for conifer morphogenesis - NAA 1-naphthaleneacetic acid     

Direct rooting and hardening of tea microshoots in the field

Micropropagation of tea (Camellia sinensis (L.) O. Kuntze) has been widely attempted but commercial exploitation of this method is limited by heavy losses during the hardening procedures. In the present study, optimization of time of harvesting (spring and early summer) of microshoots, shoot size, soil pH (4.0–6.4), plant growth regulator treatment (IBA; 500 mg l^-1, 30 min) CO₂ (9.09/10×10⁻⁵ mol l^-1 to 10.22/10×10^-5 mol l^-1 and 20/11×10⁻⁵ mol l^-1 to 80/13×10⁻⁷ mol l^-1) enrichment and light (15 μ mol m^-2 s^-1) conditions in specially designed hardening chambers, made a significant impact on the percent of success for hardening. Following the standardized procedure, up to 71.6% root induction and 73% survival could be achieved. Successful field transfer was also accomplished. This revised version was published online in June 2006 with corrections to the Cover Date.     

Development of photoautotrophy and photoinhibition of Gardenia jasminoides plantlets during micropropagation

This paper reports on the fast fluorescence responses of Gardenia jasminoides Ellis plantlets, at two successive stages (shoot multiplication and root induction) of culture in vitro. We test whether plantlets in vitro suffer photoinhibition during culture and whether the degree of photoautotrophy of these mixotrophic plantlets has any effect on the extent of photoinhibitory impairment. In this regard the effects of different sucrose levels in the medium and PPFD during growth on the development of photoautotrophy and the extent of photoinhibition were evaluated. Plantlets were grown under low, intermediate, and high (50, 100, and 300 mol m^-2 s^-1) PPFD, and at 3 different sucrose concentrations (0.5, 1.5, and 3.0%, w/v) in the medium, during shoot multiplication. During root induction the same growth conditions were assayed except for the high PPFD. The development of photoautotrophy was assessed via the difference between the stable carbon isotope composition of sucrose used as heterotrophic carbon source and that of leaflets grown in vitro. Plantlets from root induction showed more developed photoautotrophy than those from shoot multiplication. For both stages the low-sucrose medium stimulated the photoautotrophy of plantlets in vitro. In addition, intermediate PPFD induced photoautotrophy during shoot multiplication. For plantlets of both culture stages at the lowest PPFD no photoinhibition occurred irrespective of the sucrose concentration in media. However, during the shoot multiplication stage chlorophyll fluorescence measurements showed a decrease in F _v /F _m and in t _1/2 as growing PPFD increased, indicating photoinhibitory damage. The decline of F _v /F _m was caused mostly by an increase in F _o , indicating the inactivation of PSII reaction centers. However plantlets growing under low sucrose showed reduced susceptibility to photoinhibition. During root induction, only plantlets cultured with high sucrose showed a decrease in F _v /F _m as PPFD increased, although t _1/2 remained unchanged. In this case, the decline of F _v /F _m was mostly due to a decrease in F _m , which indicates increased photoprotection rather than occurrence of photodamage. Therefore, growth in low-sucrose media had a protective effect on the resistance of PSII to light stress. In addition, plantlets were more resistant to photoinhibition during root induction than during shoot multiplication. Results suggest that increased photoautotrophy of plantlets reduces susceptibility to photoinhibition during gardenia culture in vitro.Abbreviations AP apparent photosynthesis - Chl total chlorophyll content - Chl a/b chlorophyll a-to-b ratio - Chl/Car total chlorophyll-to-carotenoids ratio - ¹³C ratio of ¹³C/¹²C relative to PeeDee belemnite standard - F _m maximum chlorophyll fluorescence - F _o fluorescence emission when all reaction centres are open and the photochemical quenching is minimal - F _v variable chlorophyll fluorescence (F _m -F _o ) - F _v /F _m the ratio of variable to maximum chlorophyll fluorescence, indicator photochemical efficiency of PSII - MS medium Murashige and Skoog (1962) medium - PPFD photosynthetic photon flux density - Rd dark respiration, t _1/2 the half-time of the increase from F _o to F _m - IAA indole butyric acid     

Micropropagation and ex vitro rooting of pistachio (Pistacia vera L.)

Plant Cell, Tissue and Organ Culture (PCTOC) - An effective pistachio (Pistacia vera L.) micropropagation system was developed involving rapid axillary bud proliferation and ex vitro rooting. The...     

In vitro and ex vitro rooting of Siratia grosvenorii, a traditional medicinal plant

Since a decade, the large-scale commercial production of Siratia grosvenorii plantlets is being practiced through in vitro culture of its microcuttings, but it has some drawbacks such as handling of plantlets, low transplant-survival rate, development of massive callus, low yield after transplantation, etc. An experiment has been conducted to improve the prevailing technique as well as to develop a new ex vitro technique to overcome these drawbacks. Several concentrations of naphthalene acetic acid (NAA) (0–4.0 mg/l) have been tried with the MS (Murashige and Skoog in Physiol Plant 15:473–479, 1962) basal medium containing 3% (w/v) sucrose and 4.0 g/l agar, out of which 0.1 mg/l NAA was found best in terms of smaller diameter of callus and maximum rooting and transplant survival rate. Further, use of perlite instead of agar medium also showed possibilities for future research on commercial-scale plantlet production. Ex vitro rooting technique was found superior to the in vitro one as plantlets developed through this method had lateral roots without any callus at the base of microcuttings, just like the natural root system and of course with higher root length, rooting rates, and transplant survival rate compared to the in vitro developed plantlets. Further, this technique is economical in terms of labor and time saving and gives rise to vigorous plants which ultimately bring higher yields and profits.     

Effect of N6-isopentenyladenine concentration on growth initiation in vitro and rooting of bilberry and lingonberry microshoots

Buds and shoot tips of wild bilberry (Vaccinium myrtillus L.) and lingonberry (V. vitis-idaea L.) plants were cultured on a modified MS medium containing N⁶-isopentenyladenine (2iP), 9.8–78.4 μM, in order to study the effect of the 2iP-concentration on the initiation of growth. The experiment was first performed in the autumn and repeated in the spring to determine the influence of season on growth initiation. To optimise rooting, three different rooting treatments were tested for the bilberry and lingonberry microshoots. Shoots were rooted either in vitro with 0.49 μM IBA (indole-3-butyric acid) or ex vitro, incubating microshoots in 2.07 mM KIBA-solution (potassium salt of IBA) before planting, or microshoots were planted directly on peat without exogenous auxin. The best 2iP concentration for the initiation of the growth for bilberry was 49.2 μM and for lingonberry 24.6 μM. It was observed that increasing the 2iP concentration at the growth initiation stage increased the number of brownish explants both in bilberry and in lingonberry microcultures. Spring was a considerably better time than autumn for the initiation of new growth, for both species. The results of the rooting test showed that the KIBA-treatment before planting on peat increases rooting efficiency in both bilberry and lingonberry. This revised version was published online in June 2006 with corrections to the Cover Date.     

Development of polymorphic microsatellite markers in the medicinal plant Gardenia jasminoides (Rubiaceae)

A total of 12,960 simple sequence repeats (SSR) motifs were identified in the genome of the medicinal plant Gardenia jasminoides using Illumina-based EST sequences. Among the SSRs, mono-nucleotides were the most abundant (56.7%), followed by di- (19%) and tri-nucleotides (16.4%). AG/TC (60.2%), TTC/AAG (25.8%), and TTTC/GAAA (35.9%) repeat motifs were the most abundant of the di-, tri- and tetra-nucleotide motifs, respectively. Subsequently, twenty-five allelic, polymorphic primer pairs were identified and tested in 153 individuals from five natural populations of G. jasminoides. The number of alleles per polymorphic locus (A) ranged from two to eight. Observed and expected heterozygosity varied from 0.095 to 0.857 and 0.182 to 0.832, respectively. The PIC values for each locus ranged from 0.171 to 0.792. These new polymorphic EST-SSR markers will be useful for further genetic studies on this economically important plant.     

Germination and Growth Inhibitors in Fruits of Gardenia jasminoides

Plant germination- and growth-inhibiting activities of the iridoidglycosides (geniposide and genipin gentiobioside) containedin fruits of Gardenia jasminoides Ellis were studied. They inhibitedthe germination of G. jasminoides seeds and watercress seedsand the growth of Chinese cabbage roots. In addition, genipin,which is the aglycone of the glycosides, showed a strong inhibitingeffect on the growth of Chinese cabbage roots. (Received September 10, 1982; Accepted November 6, 1982)     

A high-frequency in vitro multiplication,micromorphological studies and ex vitro rooting of Cadaba fruticosa (L.) Druce (Bahuguni): a multipurpose endangered medicinal shrub

Physiology and Molecular Biology of Plants - An efficient and reproducible in vitro propagation protocol has been established for Cadaba fruticosa (L.) Druce. Surface-sterilized nodal stem segments...     

Isolation and purification of iridoid glycosides from Gardenia jasminoides Ellis by isocratic reversed-phase two-dimensional preparative high-performance liquid chromatography with column switch technology

A two-dimensional column-switching system without sample loop trapping, where two columns were switched directly via a six-port two-position switching valve, was successfully applied for the first time to the isolation and purification of six iridoid glycosides including geniposide, gardenoside, shanzhiside, scandoside methyl ester, deacetyl-asperulosidic acid methyl ester and genipin-1-beta-D-gentiobioside from Gardenia jasminoides Ellis, a plant used in the traditional Chinese medicine. The introduction of the six-port switching valve instead of sample loop assured 100% recovery from the first dimension to the second, and the injection volumes of the second dimension could reach 20 ml. In this mode of operation, the sample size of the two-dimensional approach was more than 1.3 times that of conventional gradient methods with even less solvent consumption. And the simultaneous operations of the two dimensions allowed the cycle time to be less than 19 min, compared with that (90 min) in the gradient elution single-dimension mode of operation. All of the six isolated iridoid glycosides were isolated at high purities of over 99% with approximately 96% recoveries.