Grafting micropropagated tea [Camellia sinensis (L.) O. Kuntze] shoots on tea seedlings – a new approach to tea propagation

Tea microshoots excised from well-established multiple shoot cultures grown in vitro and 8-week-old, three- to five-leaved seedlings from a local chinery stock (Banuri-96) and UPASI-9 (from southern India) were selected as scions and root stocks, respectively, for grafting. In addition, 4-month- and 12-month-old seedlings of Banuri-96 were also used as root stocks. Cut ends of root stocks and scions were pretreated with varying concentrations of BAP and NAA for 10 min. A treatment of BAP (5 mg/l) and NAA (5 mg/l) to both scion and stocks in water renewed foliar development at a relatively early stage (40–60 days). The grafted plants were kept in hardening chambers with CO₂-enriched air. No significant difference was observed between autograft (scion and root stock of Banuri clone) and heterograft (scion of the Banuri clone and root stock of UPASI-9). Of the three types (in terms of age) of seedling-raised root stocks employed, grafts on young tea (4-month-old) performed the best (88.33%). Grafts made in early summer established relatively faster and at a high rate of success. The percentage survival of plants transferred to the field was 88.33%. Received: 21 May 1998 / Revision received: 17 December 1998 / Accepted: 15 January 1999     

Effect of soil cadmium application and pH on growth and cadmium accumulation in roots, leaves and fruit of strawberry plants (Fragaria × ananassa Duch.)

Three strawberry (Fragaria × ananassa Duch.) cultivars Rainier, Totem and Selva were grown under greenhouse conditions in a Parkhill sandy loam soil with a background DTPA-extractable Cd concentration of 0.18 mg kg^-1 and a pH of 5.1. Experimental treatments included combinations of 4 Cd applications (0, 15, 30 and 60 mg Cd kg^-1 soil) applied as CdSO₄ and 2 soil pH values 5.1 and 6.8. Both the application of Cd and pH of the soil significantly affected plant growth, yield and Cd accumulation in plant tissue anf fruit. Although roots accumulated the highest concentrations of Cd of all plant parts investigated, increased soil Cd application reduced leaf weight more than root weight. In general, yield of strawberries was decreased by an increase in amount of soil-applied Cd, however the yield response varied among cultivars. At 60 mg Cd kg^-1 soil, yield of Rainier cultivar was reduced to 17.6% of control plants. Over 90% of total Cd taken up by plants grown in Cd-treated soil accumulated in roots, regardless of the Cd level in the soil. Root Cd concentrations ranged from 2.6 mg kg^-1 (control plants) to 505.7 mg kg^-1 (Totem plants grown in soil at highest Cd and a soil pH 5.1) and were directly related to soil Cd concentrations. Cd translocation from roots to leaves and fruit was very limited, resulting in a maximum Cd concentration in root leaf tissue of 10.2 mg kg^-1. Accumulation of Cd in fruit was found to correlate well with leaf Cd, although even at the highest amount of applied Cd, fruit Cd concentration did not exceed 700 g kg^-1 of fresh weight.Contribution no. 951     

Lignans of sesame: Purification methods,biological activities and biosynthesis – A review

Lignans are a group of compounds consisting of dimers of phenyl propane units. They are found in diverse forms distributed in a variety of plants. Sesame lignans in particular are obtained from Sesamum indicum, a highly prized oilseed crop cultivated widely in many countries in the east. The plant is the main source of clinically important antioxidant lignans such as sesamin, sesamolin, sesaminol and sesamol. These lignans exhibit antihypertensive, anticancerous and hypocholesterolemic activities as well especially in humans due to which they have become compounds of tremendous research interest in recent times. Sesamin is synthesized from shikimic acid through phenylpropanoid pathway and metabolised into enterolignans which play a pivotal role in protection against several hormone related diseases. In this paper we present an overview of current status of research on sesame lignans with respect to the analytical methods employed, the biological activities and biosynthesis of sesame lignans.     

Exosporodiella gen. nov. – a new coelomycetous fungus on leaves of Phoenix from India

A new coelomycetous fungus Exosporodiella phoenixesa gen. et spec, nov., has been described and illustrated from the leaves of Phoenix from India.     

Purification and characterization of a hydroxamic acid glucoside β-glucosidase from wheat (Triticum aestivum L.) seedlings

A beta-glucosidase (EC 3.2.1.21) with a high affinity for cyclic hydroxamic acid beta-D-glucosides was purified from 48-h-old wheat (Triticum aestivum L.) seedlings. The activity occurred transiently at a high level during the non-autotrophic stage of growth, and the nature of the transient occurrence was correlated with that of 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one glucoside (DIMBOA-Glc). The glucosidase had maximum activity at an acidic pH (pH 5.5) and the purified enzyme showed a high affinity for DIMBOA-Glc, Vmax and Km being 4100 nkat/mg protein and 0.27 mM, respectively. It also hydrolyzed p-nitrophenol beta-glycosides, as well as flavone and isoflavone glucosides, but to a lesser extent. The results indicated that the primary natural substrate for the glucosidase is DIMBOA-Glc and that the enzyme is involved in defense against pathogens and herbivores in non-autotrophic wheat. The glucosidase was found to be present as oligomeric forms with a molecular mass of 260-300 kDa comprising 60- and 58-kDa monomers. The N-terminal 12-amino-acid sequences of the two monomers were identical (Gly-Thr-Pro-(Ser?)-Lys-Pro-Ala-Glu-Pro-Ile-Gly-Pro), and showed no similarity to those of other plant glucosidases. Polyacrylamide gel electrophoresis under nondenaturing condition indicated the existence of at least eight isozymes. Three cultivars of Triticum aestivum had the same zone of glucosidase activity on zymograms, but the activity zones of the Triticum species, T. aestivum L., T. spelta L. and T. turgidum L., had different mobilities.     

Wedtrilosides A and B,two new diterpenoid glycosides from the leaves of Wedelia trilobata (L.) Hitchc. with α-amylase and α-glucosidase inhibitory activities

In the ongoing research to find new diabetes constituents from the genus Wedelia, the chemical constituent of Wedelia trilobata leaves, a Vietnamese medicinal plant species used to treat type 2 diabetes mellitus, was selected for detailed investigation. From a methanolic extract, two new ent-kaurane diterpenoids, wedtrilosides A and B (1 and 2), along with five known metabolites (3–7), were isolated from W. trilobata. The chemical structures of (1–7) were assigned via spectroscopic techniques (IR, 1D, 2D NMR and HR-QTOF-MS data) and chemical methods. The isolates were evaluated for α-amylase and α-glucosidase inhibitory activities compared to the clinical drug acarbose. Among them, compounds 4, 6, and 7 showed the most potent against α-glucosidase enzyme with IC₅₀ values of 27.54 ± 1.12, 173.78 ± 2.37, and 190.40 ± 2.01 μg/mL. While moderate inhibitory effect against α-amylase was observed with compounds 6 and 7 (with IC₅₀ = 181.97 ± 2.62 and 52.08 ± 0.56 μg/mL, respectively). The results suggested that the antidiabetic properties from the leaves of W. trilobata are not simply a result of each isolated compound, but are due to other factors such as the accessibility of polyphenolic groups to α-amylase and α-glucosidase activities.     

Molecular cloning and biochemical characterization of α- and β-tubulin from potato plants (Solanum tuberosum L.)

Few studies have investigated microtubules from plants that host pathogenic fungi. Considerable efforts are underway to find an antimitotic agent against plant pathogens like Phytophthora infestans. However, screening the effects of antifungal agents on plant tubulin in vivo or using purified native microtubule in vitro is a time consuming process. A recombinant, correctly folded, microtubule-like structure forming tubulin could accelerate research in this area. In this study, we cloned full length cDNAs isolated from potato leaves using reverse-transcribed polymerase chain reaction (RT-PCR). Solanum tuberosum (Stub) α-tubulin and β-tubulin were predicted to encode 449 and 451 amino acid long proteins with molecular masses of 57 kDa and 60 kDa, respectively. Average yields of α- and β-tubulin were 2.0–3.5 mg l^?1 and 1.3–3.0 mg l^?1 of culture, respectively. The amino acids, His6, Glu198, and Phe170 involved in benomyl sensitivity were conserved in Stub tubulin. The dimerization of tubulin monomers was confirmed by western blot analysis. When combined under appropriate conditions, these recombinant α- and β-tubulins were capable of polymerizing into microtubules. Accessibility of cysteine residues of tubulin revealed that important ligand binding sites were folded correctly. This recombinant tubulin could serve as a control of phytotoxicity of selected antimitotic fungicide compounds during in vitro screening experiments.     

Chemical constituents with ɑ,β-unsaturated carbonyl group from the whole plants of Erigeron breviscapus (Vant.) Hand.-Mazz. (Asteraceae)

Phytochemical characterization of the whole plants of Erigeron breviscapus (Vant.) Hand.-Mazz. Led to the isolation of two indole derivatives (1–2), seven monoterpenoids (3–9), two sesquiterpenoids (10–11), and four pyrone derivatives (12–15). The structures of these compounds were examined with spectroscopic methods, and also by comparing with previously reported spectroscopic data. Among them, ten compounds (1–2, 6–11, 14–15) are reported from Erigeron breviscapus for the first time. In addition, three compounds (2, 8–9) are isolated for the first time from Asteraceae family. On the basis of chemical research, we also described the chemotaxonomic significance of these compounds.     

Isolation, purification, and structure of components from acidic polysaccharides of Pleurotus ostreatus (Fr.) Quél.

Isolation of an antitumor component from polysaccharide fraction A5 of some Basidiomyces was achieved by column chromatography on Sephadex G-200. A detection method based on the specific rotatory characteristics of the polysaccharide was applied to estimate components in effluent fractions from the chromatography, and it was confirmed that a series of eluates having similar specific rotation was made up of homogeneous polysaccharide. Three components (H51, H52, and H53) were isolated, in chromatographically pure state, from fraction A5. Component H51 consisted of a skeleton of beta-(1 leads to 3)-linked glucose residues, probably having branches of galactose and mannose residues, and also containing acidic sugars. Component H53 had a main structure similarly consisting of beta-(1 leads to 3)-linked glucose residues and a larger proportion of acidic sugar than H51. Component H52 was a heteropolysaccharide made up of alpha-linked galactose and mannose residues. Components H51 and H53 had a higher and a lower molecular weight, respectively, than H52. The only antitumor-active component was H51.     

Purification and Properties of α-Glucosidase and Glucoamylase from Lentinus edodes (Berk.) Sing.

An α-glucosidase and a glucoamylase have been isolated from fruit bodies of Lentinus edodes (Berk.) Sing., by a procedure including fractionation with ammonium sulfate, DEAE-cellulose column chromatography, and preparative gel electrofocusing. Both of them were homogeneous on gel electrofocusing and ultracentrifugation. The molecular weight of α-glucosidase and glucoamylase was 51,000 and 55,000, respectively. The α-glucosidase hydrolyzed maltose, maltotriose, phenyl α-maltoside, amylose, and soluble starch, but did not act on sucrose. The glucoamylase hydrolyzed maltose, maltotriose, phenyl α-maltoside, soluble starch, amylose, amylopectin, and glycogen, glucose being the sole product formed in the digests of these substrates. Both enzymes hydrolyzed phenyl a-maltoside into glucose and phenyl α-glucoside. The glucoamylase hydrolyzed soluble starch, amylose, amylopectin, and glycogen, converting them almost completely into glucose. It was found that β-glucose was liberated from amylose by the action of glucoamylase, while α-glucose was produced by the α-glucosidase.

Maltotriose was the main α-glucosyltransfer product formed from maltose by the α-glucosidase.     

Cytotoxic constituents from the leaves of Jerusalem artichoke (Helianthus tuberosus L.) and their structure–activity relationships

Bioassay-directed phytochemical study of the Jerusalem artichoke (Helianthus tuberosus L.) leaves led to the isolation of a new sesquiterpene lactone of 3-Hydroxy-8β-tigloyloxy-1,10-dehydroariglovin (1), ten known sesquiterpene lactones (2–11) and two known flavones (12–13). Their chemical structures were elucidated on the basis of NMR (1D and 2D) and mass spectroscopic analysis. The cytotoxic activities of those compounds were subsequently tested against the MCF-7, A549 and HeLa cancer cells lines. The results indicated that sesquiterpene lactones 1–11 exhibited consistent cytotoxicity against all three cancer cell lines, while flavones 12 and 13 showed selective inhibitory activity against HeLa cell lines. Among them, compound 3 exhibited significant growth inhibitory activity against all three cell lines. The IC₅₀ values of compound 3 against MCF-7, A549 and HeLa were 1.97 ± 0.04, 7.79 ± 0.44, 9.87 ± 0.20 μg/ml, respectively. In addition, some structure–activity relationships of these sesquiterpene lactones for cytotoxicity were explored and summarized in this study.     

α-1,4-glucan phosphorylase forms from leaves of spinach (Spinacia oleracea L.)

Peptide patterns and immunological properties of the cytoplasmic and chloroplastic -1,4-glucan phosphorylase (EC 2.4.1.1) from spinach leaves have been studied and were compared with those of phosphorylases from other sources. The two spinach leaf phosphorylases were immunologically different; a limited cross-reactivity was observed only at high antigen or antibody concentrations. Peptide mapping of the two enzymes resulted in complex patterns composed of more than 20 fragments; but no peptide was electrophoretically identical in both proteins. Approximately 13 to 15 of the fragments exhibited antigeneity but no cross-reactivity of any peptide was observed. Therefore, the two compartment-specific phosphorylase forms from spinach leaves represent isoenzymes possessing different primary structures. Peptide patterns of potato tuber and rabbit muscle phosphorylase were different from those of the two spinach leaf enzymes. Although the potato tuber phosphorylase resides in the plastidic compartment and is kinetically closely related to the chloroplastic spinach enzyme, it reacted more strongly with the anti-cytoplasmic-phosphorylase immunoglobulin G. Similar results were obtained with rabbit muscle phosphorylase. These observations support the assumption that the chloroplast-specific phosphorylase isoenzyme has a higher structural diversity than does the cytoplasmic counterpart.Abbreviations EDTA ethylenediaminetetraacetic acid - Hepes 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid - PEG polyethylene glycol (approx. MW 8000) I=Schächtele and Steup 1986     

The Ethnobotany ofPluchea carolinensis (Jacq.) G. Don (Asteraceae) in the Botánicas of Miami, Florida

2006 Botánicas, or herbal shops that cater to ethnic populations, are common in culturally diverse southern Florida. One of the most frequently found plants in these botánicas isPluchea carolinensis. This small shrub in the Asteraceae is a common species in disturbed sites in the region, especially in back mangrove habitats. The binomialP. symphytifolia has been misapplied toP. carolinensis and is considered a synonym forNeurolaena lobata. Both species have similar uses and common names.Pluchea carolinensis is called salvia in Latino botánicas and la choige in Haitian botánicas in Miami. Interviewees at 17 herbal shops provided information on the uses of the plant. The most frequently mentioned ailments treated withP. carolinensis were sore throats and catarrh. The plant also is employed to alleviate headaches, stomach ailments, flatulence, high blood pressure, rash, toothaches, and it is considered to be an antiseptic, anticoagulant, and anti-inflammatory. Teas and compresses were the most common mode of preparation. The medicinal uses ofP. carolinensis in southern Florida are similar to those reported elsewhere in the Caribbean and similar to the uses of otherPluchea species. The plant also plays a role in limpiezas or spiritual cleansings and is associated with the Santería deity Babalú Ayé. This paper was Contribution 101 in Florida International University’s “Tropical Biology Publication” series.     

Remobilization of nitrogen from senescing leaves of pigeonpea (Cajanus cajan (L.) Millsp.): genotypic differences across maturity groups?

Remobilization of life nitrogen during the seed filling stage was investigated in relation to patterns of leaf abscission with three pigeonpea genotypes (Cajanus cajan L.) of different maturity duration [extra-short (ESD), short (SD), and medium (MD)].Leaflet abscission (trifoliate leaf) started from the bottom of the plants. The life span of defined leaf layers in the canopy differed among the genotypes and tended to be longer toward the top of the plants. At harvest, the leaf layer close to the pod-bearing top of the plant had a survival rate of 75% and 31% in ESD and SD pigeonpea, respectively, indicating that a large number of leaves in ESD was not entirely exploited for nutrient redistribution to the seed.Net remobilization of nitrogen from leaves during the reproductive stage was obtained from an above-ground plant budget for N and amounted to 35%, 47%, and 37% of the pod's requirement for N in ESD, SD, and MD, respectively. The amount of nitrogen in the defined leaf layers decreased exponentially with time, and the rate of N loss was calculated from the regressions in terms of half-life. For most of the layers half-life was longest in ESD pigeonpea indicating slower abscission and remobilization compared to both other genotypes.The present study compares two pigeonpea hybrids (ESD and SD) with a conventional genotype (MD). The results imply (1) that the efficiency to remobilize leaf nitrogen for seed development is related to the pattern of leaf abscission in pigeonpea, and (2) that SD pigeonpea remobilizes leaf N more efficiently than ESD and MD.     

Quantitative trait loci (QTLs) analysis of palm oil fatty acid composition in an interspecific pseudo-backcross from Elaeis oleifera (H.B.K.) Cortés and oil palm (Elaeis guineensis Jacq.)

We chose an Elaeis interspecific pseudo-backcross of first generation (E. oleifera × E. guineensis) × E. guineensis to identify quantitative trait loci (QTLs) for fatty acid composition of palm oil. A dense microsatellite linkage map of 362 loci spanned 1.485 cM, representing the 16 pairs of homologous chromosomes in the Elaeis genus from which we traced segregating alleles from both E. oleifera and E. guineensis grandparents. The relative linear orders of mapped loci suggested the probable absence of chromosome rearrangements between the E. oleifera and E. guineensis genomes. A total of 19 QTL associated to the palm oil fatty acid composition were evidenced. The QTL positions and the species origin as well as the estimated effects of the QTL marker alleles were in coherence with the knowledge of the oil biosynthesis pathway in plants and with the individual phenotypic correlations between the traits. The mapping of chosen Elaeis key genes related to oleic acid C18:1, using intra-gene SNPs, supported several QTLs underlying notably FATA and SAD enzymes. The high number of hyper-variable SSR loci of known relative linear orders and the QTL information make these resources valuable for such mapping study in other Elaeis breeding materials.     

Purification and enzymatic properties of a peroxidase from leaves of Phytolacca dioica L. (Ombú tree)

A peroxidase (PD-cP; 0.47 mg/100 g leaves) was purified from autumn leaves of Phytolacca dioica L. and characterized. PD-cP was obtained by acid precipitation followed by gel-filtration and cation exchange chromatography. Amino acid composition and N-terminal sequence of PD-cP up to residue 15 were similar to that of Spinacia oleracea (N-terminal pairwise comparison showing four amino acid differences). PD-cP showed a molecular mass of approx. 36 kDa by SDS-PAGE, pH and temperature optima at 3.0 and 50.0°C, respectively and seasonal variation. The Michaelis-Menten constant (K(M)) for H(2)O(2) was 5.27 mM, and the velocity maximum (V(max)) 1.31 nmol min(-1), while the enzyme turnover was 0.148 s(-1). Finally, the presence of Ca(2+) and Mg(2+) enhanced the PD-cP activity, with Mg(2+) 1.4-fold more effective than Ca(2+)     

Coumarins from the stems and leaves of Clausena dunniana H. Lév

Phytochemical study on the stems and leaves of Clausena dunniana H. Lév. led to the isolation and identification of 14 coumarins (1–14). Their chemical structures were determined on the basis of MS, NMR, and further supported by comparison with those reported in the literature. This is the first report that compounds 1–2, 4, 8, and 12–14 are present in the genus Clausena, and all of these compounds were isolated from C. dunniana for the first time. The chemotaxonomic significance of these isolated compounds was discussed.     

Morphological, physiological, cytological and phytochemical studies in diploid and colchicine-induced tetraploid plants of Echinacea purpurea (L.)

Echinacea purpurea (L.) is one of the important medicinal plant species. To obtain the tetraploid plants of Echinacea purpurea with improved medicinal qualities, the root tips of two true leaves seedlings were imbibed in 0.25 % (w/v) colchicine solution for 24, 48, 72, 96 and 168 h. The ploidy level of plants was determined by chromosome counting of root tip cells, and confirmed by flow cytometric analysis. Tetraploid induction occurred in seedlings treated for 24, 48 and 72 h at colchicine solution. The morphological, physiological, cytological, and phytochemical characteristics of diploid and colchicine-induced tetraploid plants were compared. Results indicated that tetraploid plants had considerable larger stomata, pollen grain, seed and flower. Moreover, chloroplast number in guard cells, amount of chlorophyll (a, b, and a + b), carotenoids as well as width and thickness of leaves were increased in tetraploids. However, stomata frequency, leaf index, plant height, and quantum efficiency of photosystem II in tetraploid were lower than diploid plants. High-performance liquid chromatography analysis showed that leaves of the tetraploid plants had more cichoric acid (45 %) and chlorogenic acid (71 %) than diploid plants. It was concluded that morphological and physiological characteristics can be used as useful parameters for preliminary screening of putative tetraploids in this species.     

Micropropagation of ‘Holy Basil’ (Ocimum sanctum Linn.) from young inflorescences of mature plants

In vitro micropropagation of holy basil (Ocimum sanctum L.), an Indian medicinal herb, has been accomplished on Murashige and Skoog (MS) medium utilizing young inflorescence explants. MS supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) or thidiazuron (TDZ) produced only non-morphogenetic callus. Direct multiple shoots differentiated within 2--3 weeks when explants were cultured on MS containing 6-benzyl aminopurine (BAP). Of the various levels of BAP tested, MS + BAP (1.0 mgl^–1) produced the maximum number of shoots. Incorporation of indole-3-acetic acid (IAA) (0.05 mgl^–1) along with BAP (1.0 mgl^–1) in the culture medium showed a marked increase in the number of shoots. About 92% of the in vitro regenerated shoots rooted on MS hormone-free medium within 2--3 weeks of culture and 85% of the micropropagated plantlets could be successfully established in soil, where they grew normally.     

Partial purification and characterization of the soluble DNA polymerase (polymerase-α) from seedlings of Pisum sativum L.

Radish seedlings (Raphanus sativus L. Saxa Treib) were grown in the dark with or without added kinetin (2 mg/l=9.29 M). Low-temperature (77°K) fluorescence emission and absorption spectra of etiolated cotyledons were registered at increasing seedling age before and immediately, 30 s and 30 min after one 1-ms flash. Kinetin was found to induce a higher accumulation of the phototransformable protochlorophyll(ide) P_657–650 in the etiolated cotyledons, especially from day 6 to day 10 after germination. The amount of the P_657–650 protochlorophyll(ide) resynthesized during a 30-min dark period after a 1-ms flash decreased with seedling age. It was smaller in cotyledons from kinetin-treated seedlings at day 6 after germination and at that age only. The ability to perform the Shibata shift decreased with increasing seedling age. In cotyledons from 10- and 13-day-old seedlings, the shift was accomplished to a greater extent when the plants were grown in the presence of kinetin.