Effects of Seselin and Coumarin on Growth, Indoleacetic Acid Oxidase, and Peroxidase, with Special Reference to Cucumber (Cucumis sativa L.) Radicles

Seselin, a natural coumarin derivative isolated from citrus roots, inhibited radicle growth in seedlings of cucumber (Cucumis sativa), lettuce (Lactuca sativum), radish (Raphanus sativus), and wheat (Triticum aestivum) grown in the dark. Coumarin similarly inhibited radicle growth of cucumber seedlings. Growth retardation of the cucumber radicles was accompanied by an increased activity of peroxidase and indole-3-acetic acid oxidase. Both compounds antagonized indole-3-acetic acid-induced growth of wheat coleoptiles, whereas coumarin was much less effective than seselin in antagonizing gibberellic acid-induced release of reducing sugars from barley endosperm. It is suggested that seselin plays an important role in the regulation of root growth, and that it is the indole-3-acetic acid oxidase cofactor previously detected in citrus roots.     

Light activation of vindoline biosynthesis does not require cytomorphogenesis in Catharanthus roseus seedlings

Upon illumination, the cotyledons of Catharanthus roseus seedlings readily synthesise vindoline from late biosynthetic intermediates, which accumulate in etiolated seedlings. The cellular localisation of tryptophan decarboxylase (TDC) and desacetoxyvindoline 4-hydroxylase (D4H), which catalyse the first and penultimate reactions of vindoline biosynthesis, was identified by immunocytochemistry in developing seedlings. The expression of TDC was restricted to the upper epidermis of cotyledons, whereas that of D4H was confined to laticifer cells. Light exposure of etiolated seedlings significantly induced D4H enzyme activity without changing the steady-state levels of D4H immunoreactive protein or modifying the cellular distribution of D4H expression in dark-grown seedlings. These results suggest that the early and late stages of vindoline biosynthesis occupy different cellular compartments, even in the early phases of etiolated seedling development. The role of light in activating the late stages of vindoline biosynthesis does not, therefore, seem to be related to the formation of the laticifer and idioblast cell types. It is concluded that light is not required for formation of these cell types, whereas regulatory factors, restricted to idioblasts and laticifers, may respond to light to activate localised expression of the late stages of vindoline biosynthesis.     

STUDIES ON THE GERMINATION OF SEEDS OF THE ROOT PARASITES,ALECTRA VOGELII AND STRIGA GESNERIOIDES. I. ANATOMICAL CHANGES IN THE EMBRYOS

Anatomical changes in the radicle and shoot meristems of embryos of germinating seeds of the obligate root parasites, Alectra vogelii and Striga gesnerioides were studied. When germination of seeds was stimulated by cowpea (Vigna unguiculata) root exudate, growth occurred mainly in the radicular pole of embryos and minimally in the plumular pole, resulting in seedlings with elongated radicles. Maximum radicle elongations of about 3 mm in A. vogelii and 2 mm in S. gesnerioides were recorded during a period of 8 and 11 days, respectively. Analysis of the radicular tip during the course of seed germination revealed that the activity of the meristematic tissue progressively decreased until it completely disappeared. When germinated seeds were cultured on nutrient agar media, the radicle meristem of A. vogelii continued to grow producing a normal root with a root cap. On the other hand, the radicles of cultured S. gesnerioides seeds elongated only slightly before meristematic activity ceased. During continued growth of seedlings of both species on agar media, lateral roots whose tips had typical angiosperm root topography, were initiated from the radicle.     

Developmental Regulation of Enzymes of Indole Alkaloid Biosynthesis in Catharanthus roseus

Developing seedlings of Catharanthus roseus were analyzed for appearance of tryptophan decarboxylase (TDC), strictosidine synthase (SS), N-methyltransferase (NMT) and O-acetyltransferase (DAT) enzyme activities. SS enzyme activity appeared early after germination and was present throughout most of the developmental study. TDC activity was highly regulated and peaked over a 48 hour period achieving a maximum by day of 5 of seedling development. Both TDC and SS were present in all tissues of the seedling. NMT and DAT enzyme activities were induced after TDC and SS had peaked and these activities could only be found in hypocotyls and cotyledons. TDC, SS, and NMT did not require light for induction whereas DAT enzyme activity was increased approximately 10-fold after light treatment of dark grown seedlings.     

Annuario della Società Botanica Italiana 1932

A sandy culture experiment was conducted to investigate the effects of exogenous CaCl₂ on the indole alkaloid accumulation in Catharanthus roseus under salt stress. One-month seedlings of C. roseus were treated with the different concentrations of NaCl (0, 50, and 100 mmol l^? 1) and 7.5 mmol l^? 1 CaCl₂. The plant samples were analyzed after 7 days of the treatments. The NaCl-stressed plants showed decrease of fresh and dry weight and increase of malondialdehyde (MDA) content compared to control. Tryptophan decarboxylase (TDC) activity increased significantly under 50 mmol l^? 1 NaCl without CaCl₂ addition, 50 mmol l^? 1 NaCl with 7.5 mmol l^? 1 CaCl₂, and CaCl₂ treatment without NaCl addition. There was a significant increase in peroxidase activity under NaCl stress compared to control. The vindoline, catharanthine, vincristine, and vinblastine contents increased under salt stress (especially with 50 mmol l^? 1 NaCl treatment with or without CaCl₂). Addition of CaCl₂ to NaCl-stressed plants increased biomass, TDC activity, vindoline, and catharanthine contents and lowered MDA and vincirstine contents compared to the plants without CaCl₂. The plants treated with CaCl₂ alone showed higher TDC activity, vindoline, catharanthine, and vinblastine content when compared to control. The results showed that exogenous CaCl₂ could promote the indole alkaloid metabolism under salt stress.     

High levels of tryptamine accumulation in transgenic tobacco expressing tryptophan decarboxylase

A full-length complementary DNA clone encoding tryptophan decarboxylase (TDC; EC 4.1.1.28) from Catharanthus roseus (De Luca V, Marineau C, Brisson N [1989] Proc Natl Acad Sci USA 86: 2582-2586) driven by the CaMV 35S promoter was introduced into tobacco (Nicotiana tabacum) to direct the synthesis of the protoalkaloid tryptamine from endogenous tryptophan. Young, fully expanded leaves of CaMV 35S-TDC transformed plants had from four to 45 times greater TDC activity than did controls. Tryptamine accumulated in transgenic plants to levels that were directly proportional to their TDC specific activity. Despite their increased tryptamine content, the growth and development of the CaMV 35S-TDC plants appeared normal with no significant differences in indole-3-acetic acid levels between high tryptamine and control plants. Plants with the highest TDC activity contained more than 1 milligram of tryptamine per gram fresh weight, a 260-fold increase over controls.     

The effect of phytohormones produced byArthrobacter sp. on the phosphatase activity in plant roots

Phytohormonal activity (auxins, gibberellins, cytokinins) was tested in the supernatant of a culture ofArthrobacter sp. Crude extract of the phytohormonal fraction was used as substrate for the growth of Lactuca sativa seedlings. Treating with bacterial hormones resulted in an increased plant development. Furthermore, a sharp increase of the acid phosphatase activity was observed in the roots.     

盐胁迫对长春花幼苗生长和生物碱含量的影响

以NaCl浓度分别为0、50、100、150、200和250 mmol·L^-1的1/2 Hoagland营养液处理长春花幼苗,7 d后测定其鲜质量、干质量、丙二醛(MDA)和叶绿素含量、色氨酸脱羧酶（TDC）和过氧化物酶（POD）活性等生理指标及文多灵、长春质碱、长春新碱和长春碱等生物碱含量.结果表明：NaCl显著地降低长春花幼苗的鲜质量和干质量,提高MDA含量;叶绿素含量在低盐浓度(50 mmol·L^-1)下与对照相比差异不显著,在高于50 mmol·L^-1时随NaCl浓度的增加而逐渐降低;在NaCl处理下,POD活性与对照相比显著上升;TDC活性在50 mmol·L^-1 NaCl处理下活性最高,而后随盐浓度的增加逐渐降低;文多灵、长春质碱、长春新碱和长春碱含量都是在50 mmol·L^-1NaCl处理下最高,分别为4.61、3.56、1.19和2.95 mg·g^-1,并显著高于对照及其他处理.盐胁迫虽然在一定程度上抑制了长春花幼苗生长,但促进了其生物碱的代谢,提高了生物碱含量;50 mmol·L^-1NaCl处理对长春花吲哚生物碱代谢的促进作用最大.     

Epicotyl morphophysiological dormancy in seeds of Lilium polyphyllum (Liliaceae)

Dormancy-breaking and seed germination studies in genus Lilium reveal that the majority of Lilium spp. studied have an underdeveloped embryo at maturity, which grows inside the seed before the radicle emerges. Additionally, the embryo, radicle or cotyledon has a physiological component of dormancy; thus, Lilium seeds have morphophysiological dormancy (MPD). A previous study suggested that seeds of Lilium polyphyllum have MPD but the study did not investigate the development of the embryo, which is one of the main criteria to determine MPD in seeds. To test this hypothesis, we investigated embryo growth and emergence of radicles and epicotyls in seeds over a range of temperatures. At maturity, seeds had underdeveloped embryos which developed fully at warm temperature within 6 weeks. Immediately after embryo growth, radicles also emerged at warm temperatures. However, epicotyls failed to emerge soon after radicle emergence. Epicotyls emerged from >90% seeds with an emerged radicle only after they were subjected to 2 weeks of cold moist stratification. The overall temperature requirements for dormancy-breaking and seed germination indicate a non-deep simple epicotyl MPD in L. polyphyllum.     

Tryptophan decarboxylase from transformed roots of Catharanthus roseus

Tryptophan decarboxylase (TDC, EC 4.1.1.28) from Catharanthus roseus hairy roots was purified 80-fold. Antibodies against TDC were obtained and they recognized only one protein of 55 kDa in crude extracts from hairy root cultures. Elicitation of transformed root cultures with macerozyme yielded a marked increase in TDC activity, which was accompanied by a similar increase in the amount of immunoreactive TDC protein. These results suggest that the alkaloid accumulation, produced by elicitation, requires the synthesis of new TDC polypeptide in C. roseus root cultures and establishes important differences in the regulatory control of this enzyme in root cultures compared to developing seedlings, where the posttranslational regulation apparently plays a major role.     

The Influence of Auxin and Ethylene on Chromatin-directed Ribonucleic Acid Synthesis in Soybean Hypocotyl

Soybean seedlings treated with ethylene exhibited small increases in ribonucleic acid content in the elongating section of the hypocotyl. Chromatin isolated from the elongating section of ethylene-treated seedlings showed a 35 to 60% increase in the capacity for RNA synthesis. The ethylene-induced response was saturated at 1 microliter/liter of ethylene and was fully expressed after 3 hours. Auxin caused marked accumulation of RNA and DNA in the elongating and basal tissue of the hypocotyl. Chromatin isolated from these auxin-treated tissues showed an 8- to 10- fold increase in RNA synthetic capacity as measured in vitro. Ethylene added with auxin reduced the auxin enhancement of nucleic acid synthesis in the elongating and basal tissues. Both ethylene and auxin treatment of the seedlings inhibited nucleic acid accumulation and chromatin activity in the apical tissue. Ethylene did not appear to mediate the auxin effects on nucleic acid synthesis in soybean hypocotyl with the possible exception of inhibition in the apical tissue.     

The HPLC profile of proteins of thermoprotection-acquired wheat (Triticum aestivum L.) seedlings

A pre-treatment of 40 °Cprovided thermoprotection to wheat seedlings against 43 °C, which was otherwise a lethal temperature. Due to temperature pretreatment, the rate of protein synthesis at 45 °C increased in both plumules and radicles. The HPLC profile of plumule and radicle proteins of thermoprotection-acquired seedlings was different from the plumules and radicles of non-treated seedlings.     

Stress-associated factors increase after desiccation of germinated seeds of <Emphasis Type="Italic">Tabebuia impetiginosa</Emphasis> Mart.

Improved re-establishment of desiccation tolerance was studied in germinated seeds of Tabebuia impetiginosa Mart. by exposing to a polyethylene glycol solution prior to desiccation. The effects of different osmotic potentials and drying rates were studied. In addition, temporary temperature stress and exogenous abscisic acid were applied to evaluate their effect on desiccation tolerance of the protruded radicle. An osmotic potential of −1.7 MPa at 5°C followed by slow drying was most effective in the re-establishment of desiccation tolerance in protruded radicles with a length up to 3 mm. An osmotic potential of −1.4 or −2.0 MPa was less effective. Fast drying completely prevented the re-induction of desiccation tolerance. Cold shock or heat shock prior to osmotic treatment as well as abscisic acid added to the osmotic solution improved desiccation tolerance of protruded radicles. Surprisingly, survival of the germinated seed did not depend on re-establishment of desiccation tolerance in the protruded radicle. Even after the protruded radicle became necrotic and died, the production of adventitious roots from the hypocotyls allowed for survival and the development of high quality seedlings. Thus, T. impetiginosa appeared to be well adapted to the seasonally dry biome in which the species thrives via mechanisms that offer protection against desiccation in the young seedling stage.     

Analysis of In Vivo Protein Synthesis and Histological Studies of Haustorial Formation in Root Cultures of Witch weed (Striga asiatica L Kuntze)

We recently described an in vitro approach that uses root culturesto study haustorial formation in Striga asiatica. Previous studieshave used haustoria formed on intact radicles of Striga seedlings.In vitro cultured roots formed haustoria that appeared morphologicallysimilar to those formed by Striga radicles, but were 5–10-foldlarger. In this study, we provide biochemical and histologicalevidence to support further the similarity of root culture haustoriato haustoria formed on radicles of seedlings. We examined invivo protein synthesis during haustorial development on rootcultures and radicles by 2-D PAGE. Four proteins increased inabundance in both root cultures and radicles after 6 h of haustorialinduction. All four proteins appeared transiently in root culturesand radicles, being more abundant at 6 h, and less abundantafter 24 h of haustorial induction. Only three of the four haustorial-specificproteins were more abundant in root cultures after 2 h of haustorialinduction; all four had decreased in abundance after 12 h ofhaustorial induction. Using light microscopic analysis we comparedthe ontogeny of root culture haustoria to that of haustoriaon radicles. These studies revealed that root culture haustoriaundergo developmental changes similar to those reported forradicle haustoria such as early expansion of cortical cells,the emergence of haustorial hairs from epidermal cells, andthe development of densely staining cells at the haustorialapex. In addition, these changes occurred within a similar time-frameand sequence in root culture and radicle haustoria. Finally,root culture haustoria were found to be capable of attachingto sorghum host roots. Key words: Striga asiatica L., Kuntze, haustoria, root cultures, proteins, histology, 2D-PAGE     

Casbene: An anti-fungal diterpene produced in cell-free extracts of Ricinus communis seedlings

The capacity of cell-free extracts of 2·5-day-old castor bean seedlings for synthesis of casbene from mevalonic acid were compared for seedings which had been germinated under sterile conditions and seedlings which were intentionally exposed to fungal cultures. Extracts from seedlings exposed to cultures of Rhizopus stolonifer, Aspergillus niger or Fusarium moniliforme produced much higher levels of casbene than extracts from sterile controls; the initial rates of casbene synthesis were 20–40 times higher in the extracts of seedlings that had been exposed to fungus. Although some variation in the capacity for synthesis of other diterpene hydrocarbons from mevalonic acid was seen in the two types of extracts, no consistent or striking stimulation in the synthesis of any of these was noted under these conditions of exposure of the seedlings to fungi. The potato-dectrose agar used as a fungal growth medium did not itself evoke the increase in casbene synthesis. Intact mycelia and cell-free extracts of mycelia of Rhizopus stolonifer gave no indication of diterpene biosynthesis from mevalonic acid. Purified casbene at concentrations of 10 μg ml⁻¹ or greater retarded the development of A. niger on potato-dextrose medium. Casbene was also found to inhibit the endogenous and gibberellic acid-stimulated growth of leaf sheaths of the dwarf-5 mutant of Zea mays and of the growth of the K-12 strain of Escherichia coli on glucose-minerals medium. It is suggested that casbene may serve the castor bean plant as a phytoalexin.     

A smoke-derived butenolide improves early growth of tomato seedlings

3-Methyl-2H-furo[2,3-c]pyran-2-one is an active compound isolated from plant-derived smoke water. It has a stimulatory role during seed germination similar to that of smoke or aqueous extracts of smoke. The present study was undertaken to gain insight into the physiological events involved in seed germination and seedling development and which are affected by butenolide using tomato (Lycopersicon esculentum Mill.) cultivar “Heinz 1370” seeds. No stimulatory role on the seed germination of tomato was recorded following the use of the butenolide, however, post-germinative growth of tomato seedlings was significantly improved over the control (P ≤ 0.05). The emergence of the radicle and elongation of the hypocotyls and radicles were accelerated in seeds imbibed with butenolide at 10⁻⁷ M. Flow cytometry studies showed that in butenolide-treated seeds the ratio of cells with replicated DNA was increased. Seedling vigour and weight were significantly increased by the butenolide (P ≤ 0.05). An inverse correlation was observed between the weight of cotyledons and the weight of the hypocotyls and radicle during seedling development. This is an indication that the butenolide is implicated in mobilization and utilization of stored reserve materials in developing tomato seedlings.     

Interactive effects of exogenous growth regulators and artificial acid rain on cytokinins and gibberellins in maple seedlings

Vegetation experiments were conducted under greenhouse conditions in which long-term effects of simulated acid rain on the shoots, and short-term effects of acidified root medium on the roots of two maple species (Acer pseudoplatanus L. andAcer negundo L.) were studied. Both alternatives of the acidification stress were studied with respect to exogenous applications of auxins and gibberellins. Changes in seedling growth caused by acid rain were not marked, but changes in the levels of endogenous growth regulators were detected, both the long-term exposure ofAcer pseudoplatanus L. seedling shoots to acid rain and the short-term acidification of root environment resulted in enhanced cytokinin activity and in decreased gibberellin activity. But when A.pseudoplatanus L. seedlings were pretreated with gibberellins, their activity was further increased by increased acidity.Acer negundo L. seedlings responded to the acidification of their root environment with enhanced cytokinin activity only when pretreated with exogenous auxins. The differences in the responses ofA. pseudoplatanus L. and A.negundo L. to the acidification can be explained in terms of their distinct physiological properties, including changes recorded in chlorophyll content. The results obtained show that a pretreatment of maple seedlings with exogenous growth regulators can modify the effects of increased acidity in the environment on their physiological status as characterized by the level of endogenous cytokinins and gibberellins.     

Hormonal Relations in the Phototropic Response IV. Light-Induced Changes of Endogenous Auxins in the Coleoptile

Beside indoleacetic acid (IAA), 3 auxins were found by chromatographic resolution of acidic fractions of Avena and Zea coleoptile tips. One of these auxins, designated P, occurred at levels of activity approaching those of IAA. The other 2 auxins, termed F and M, occurred at lower levels of activity. When the auxins of the excised coleoptile tips were isolated immediately after equilateral or unilateral irradiation with blue light at first positive energies, the ratio of IAA to the other auxins increases. This rise is the result of a decrease in P and F, and probably an increase in IAA. Light did not affect materially the total auxin content. It is suggested that P and F might be associated with the basipetal transport inequalities of IAA in phototropism.

P has been partially characterized. Its R_F on chromatograms developed in ammoniacal isopropanol is about 0.65. It is converted to IAA in vitro by heat. The ultraviolet absorption spectrum of chromatographically resolved P also suggests an indolyl complex. P is not readily transported basipetally, and the slope of its relative concentration-response curve (Avena section test) is lower than that of IAA. P does not appear to be any of the chemically characterized native auxins.