Lipolytic and peroxidative enzyme expressions in cultured potato tuber cells

Potato cells (cv. Norchip) were cultured from tuber parenchymal tissue and subcultured to dissociate and habituate the despecialized cells. After several subculturings on a minimal nutrient media, this line of cells demonstrated repeatable physical growth profiles for dry weight (DW), fresh weight (FW) and protein. Two enzymes of plant lipid metabolism were investigated, lipolytic acyl hydrolase (LAH) and lipoxygenase (LOX), which respectively liberate and peroxidize fatty acids from lipid in cellular membranes. LAH, measured as p-nitrophenyl palmitate hydrolase, was present in this line of cells in easily detectable amounts (317 units g^-1 DW) albeit much lower than that found in mother tuber (9878 units g^-1 DW). The presence of LAH in this line is significant because LAH isozymes are often described as storage proteins, yet activity per gram fresh weight in these unorganized cells is reasonably constant until culture growth exits the linear phase. However, LOX, the most active free fatty acid metabolizing enzyme in potato tubers (89,800 units g^-1 DW), was not detectable in this line of callus or suspension cultured cells. The absence of LOX activity in this line of cells was verified by a number of assay approaches and was confirmed by activity staining of extracted enzymes separated in polyacrylamide gels. The absence of LOX in these cultured cells is especially important in determining the functions of this lipid peroxidation system and how it may be genetically regulated.Mention of company or trade name does not imply endorsement by the United States Department of Agriculture over others not named.A laboratory cooperatively operated by the Midwest Area, Agricultural Research Service, U.S. Department of Agriculture, The Minnesota Agricultural Experiment Station, the North Dakota Agrcultural Experiment Station, and the Red River Valley Potato Grower's Association.     

Growth and carbon utilization by sprouted propagules of two species of submersed rooted aquatic plants grown in darkness

Hydrilla verticillata (L. f.) Royle tubers from monoecious plants andPotamogeton gramineus L. winter buds were sprouted and allowed to grow in the dark for 120 days. We measured plant length and counted the number of leaves at 2–3 day intervals.Hydrilla grew most rapidly during the first 16–17 days andPotamogeton grew most rapidly during the first 16–25 days. Measurement of propagule carbon content over time indicated that cessation of rapid growth coincided with depletion of tuber carbon by one-half forHydrilla. ForPotamogeton, growth was reduced after 16 to 25 days while the winter bud C half-life was 37 days. Calculations indicated thatHydrilla mobilized 49% andPotamogeton 39% of the initial propagule carbon to support growth. In a second experiment, in which plants were grown in substrate the plants grew taller and produced slightly more leaves per plant.Potamogeton removed from darkness after specified time periods, and allowed to grow for 21 days in a greenhouse recovered from 20–30 days in the dark. Similarly treatedHydrilla plants recovered from up to 80 days in the dark.Potamogeton had mobilized 79% of initial C by the time it was unable to recover from the dark treatment. Combined results for both species indicate that the majority of propagule C was utilized in the first 16 to 30 days following sprouting. In conjunction with an understanding propagule sprouting requirements, this information will be useful in the timing of application for management techniques. The U.S. Government right to retain a non-exclusive, royalty-free licence in and to any copyright is acknowledged. The U.S. Government right to retain a non-exclusive, royalty-free licence in and to any copyright is acknowledged.     

Immunolocalization of extensin and potato tuber lectin in carrot, tomato and potato

Tissue-specific expression of two members of the cell wall hydroxyproline-rich glycoprotein (HRGP) family, extensin and potato tuber lectin, was examined by immunolocalization at the light microscope level in various organs (leaves, stems, roots, fruit, tuber) of carrot ( Daucus carota cv. Thumbelina), tomato ( Lycopersicon esclentum cv. Pixie Hybrid II), and potato ( Solanum tuberosum cv. Kennebec). Extensin was prominently expressed in vascular tissue, particularly xylem and also phloem, although virtually all cells displayed some degree of staining which varied as a function of the tissue, organ, and plant under study. Antibodies against potato tuber lectin (PTL) displayed a localization pattern similar to that observed for extensin; notably PTL did not stain cambium but did stain epithelial cells lining secretory cavities. These distribution patterns are consistent with a role for extensin, and possibly PTL, in providing mechanical support in tissues subjected to compression or torsional stress imparted by vascular growth, or by similar stress brought about by transport of vascular fluids.     

印产毛喉鞘蕊花的引种栽培研究

本文报道印产毛喉鞘蕊花肉质根稳定性状的分离及栽培技术路线．肉质根株提纯到９９．５％;求得了露地最佳效益种植法,最优水平搭配是：３０ｄ、１５株／ｍ２、高垄、覆膜、底肥、整枝、去花穗,产鲜根８７００ｋｇ／ｈｍ２．     

Properties of polyphenol oxidase from tubers of the yam Dioscorea bulbifera

The subunit MW of Dioscorea bulbifera polyphenol oxidase (MW 115 000 ± 2000) determined by SDS-PAGE is ca. 31 000 indicating that the enzyme is an oligomeric protein with four subunits. K_i values of various inhibitors and their modes of inhibition have been determined with catechol and pyrogallol as substrates. p-Nitrophenol, p-cresol, quinoline and resorcinol are competitive inhibitors of catechol binding while only orcinol and p-nitrophenol behave in the same way towards pyrogallol as substrate. From the effect of pH on V_max, groups with pK values ca. 4.7 and 6.8 have been identified to be involved in catalytic activity. The Arrhenius activation energy (E_a) at pH 4.0 is 8.9 kcal/mol between 40–65°. At pH 7.0, the value is 22.1 kcal/mol between 40 and 60°. The enthalpies (ΔH) at pH 4.0 and pH 7.0 are 2.3 kcal/mol and 32.4 kcal/mol respectively. The results are discussed considering the conformational changes of the enzyme during substrate binding.     

Purification and some properties of polyphenol oxidase from the yam tubers, Dioscorea bulbifera

In a comparison of the polyphenol oxidase activity of various species of yam tubers the greatest enzyme activity was found in D. bulbifera. The enzyme was purified from acetone powder extracts of this plant. Ammonium sulphate fractionation, followed by ion exchange chromatography and gel filtration gave 22-fold purification. The final product gave a single band on polyacrylamide disc gel electrophoresis. The purified enzyme showed activity towards catechol, pyrogallol and dl-β-3,4-dihydroxyphenylalanine (dl-DOPA) and had a MW 115000 ± 2000. It was characterized by response to various inhibitors. β-Mercaptoethanol, dithioerythritol, l-cysteine, sodium metabisulphite and KCN inhibited strongly.     

Activity of phosphorylase in Solanum tuberosum during low temperature storage

Changes in the activity of phosphorylase were measured during storage of potatoes at + 2° when the sugar content rises rapidly and subsequently at + 10° when the accumulated sugar is converted mainly to starch. The observed changes were relatively small and could not be related to any of the components of the phosphorylase system, which was shown to be complex.     

Cadmium loading into potato tubers: the roles of the periderm, xylem and phloem

The mobility of Cd in potato plants (Solanum tuberosum) was examined using both short‐term radioisotopic labelling with ¹⁰⁹Cd and long‐term growth experiments in soil supplemented with Cd, with an emphasis on the pathways through which Cd is taken up by tubers. Split‐pot experiments showed that tubers and their associated stolons and stolon roots contribute only a minor fraction to the overall Cd absorption by the plant. Most of the Cd was absorbed by the basal roots. ¹⁰⁹Cd absorbed from the soil was rapidly exported to other parts of the plant, especially the stem, with significant amounts appearing in the tubers within 30 h. Application of ¹⁰⁹Cd to leaves showed that Cd can be rapidly distributed via the phloem to all tissues. The results suggest that unlike Ca, Cd has high mobility in plants in both xylem and phloem, and that stems may have an important role in transfer between these two pathways.     

彩色马蹄莲的组织培养

以彩色马蹄莲芽眼为外植体,在添加不同激素配比的MS培养基上进行培养.结果表明:(1)较适宜诱导愈伤组织的激素组合是1.5 mg/L Zt+0.1 mg/L NAA、3.0 mg/L 6-BA+0.1 mg/L NAA或0.5 mg/L 2,4-D+0.1 mg/L NAA,出愈率分别为94.12%、100%和86.67%;较适宜诱导不定芽的激素组合是2.0 mg/L 6-BA+0.1 mg/L NAA,芽丛诱导率可达87.3%;芽增殖较适宜的激素组合是2.0 mg/L 6-BA+0.1 mg/L NAA或2.0 mg/L KT+1.0 mg/L NAA,而且在继代培养中,交替使用这两种激素组合,繁殖系数可达4倍以上;生根培养基以1/2 MS+0.3 mg/L NAA+0.2 mg/L IAA较为适宜,生根率达95%以上.(2)彩色马蹄莲小块茎形成的主要影响因素是培养基中的蔗糖浓度,最适宜蔗糖浓度为10%;10 mg/L多效唑和8 mg/L NAA对小块茎的形成有明显的促进作用,而分裂素则表现出较强的抑制作用.