Effects of Storage Temperature and Wounding on Cytokinin Levels in Potato Tubers

The level of butanol-soluble cytokinin in potato tubers (Solanumtuberosum L.) decreased greatly after harvest of the tubers,remained low during rest-period and then began to increase priorto the end of the period. In warm-stored tubers (25?C) the levelrose faster than in cold-stored ones (4?C). On the other hand,the level of water-soluble cytokinin, which was high duringthe rest-period, decreased after the rest was broken. Wounding the tubers caused an increase in butanol-soluble cytokinin.The increase in cytokinin within the first 24 hr after woundingwas smaller in warm-stored tubers than in cold-stored ones andwas decreased with increasing storage time of the tubers. Thelevel of water-soluble cytokinin decreased by wounding. Theseresults suggest that water-soluble cytokinin is a storage formand that the level of butanol-soluble cytokinin is regulatedby an interconversion between the two. (Received February 6, 1982; Accepted May 13, 1982)     

Tissue Browning of Potato Tubers Induced by Gamma Irradiation

A considerable browning was observed especially in cortex tissue and along xylem of potato tubers harvested at Sakai in Osaka Prefecture, after irradiation with 10, 20 and 50 krad doses of cobalt-60 gamma rays. This phenomenon was accompanied by the marked increase in polyphenol content and peroxidase activity, and the transient increase in o-diphenol oxidase activity. Total reducing compounds in the tissue were also increased by gamma irradiation.

The browning phenomenon depended on the storage period from the harvest to gamma irradiation treatment. The browning and the transient increase in o-diphenol oxidase activity were completely suppressed in the case of tubers irradiated 3 months after harvest.

There was no significant change in α-amylase activity in all tubers tested.     

Effects of Physiological Changes in Potato Tubers (Solanum tuberosum L.) after Low Temperature Storage on the Level of Acrylamide Formed in Potato Chips

Acrylamide in potato chips made from tubers stored at 2 or 20°C for two weeks after harvest was analyzed by GC-MS. The acrylamide level in the former chips was higher than ten times of that in the latter, which was highly correlated with both glucose and fructose levels in the tubers.     

High-Level Expression of Ice Nuclei in Erwinia herbicola Is Induced by Phosphate Starvation and Low Temperature

In laboratory cultures of ice nucleation-active (Ice⁺) Erwinia herbicola isolates, it has been difficult to achieve high-level expression of ice nuclei, especially nuclei active at temperatures warmer than −5°C (i.e., type 1 ice nuclei). Here we demonstrate that starvation for phosphate and exposure to low temperature triggers expression of ice nuclei in E. herbicola cultures. Starvation for nitrogen, sulfur, or iron was less effective. Under optimal conditions with two different strains, essentially all cells produced ice nuclei active at −10°C or warmer, with an average of 22% containing type 1 ice nuclei within 1 h of a low-temperature shift. These conditions did not greatly enhance the shedding of ice nucleation-active membrane vesicles that are known to be produced by Ice⁺ E. herbicola isolates. These results support the theory that the Ice⁺ phenotype may allow nutrient-limited epiphytes to trigger freezing damage, releasing nutrients from host plants. Received: 2 November 1997 / Accepted: 5 January 1998     

A Note on the Determination of Alcohol in Potato Tubers

Certain samples of potato tubers have been found to containan interfering substance which is estimated as alcohol by themicro-method of Friedemann and Klass (1936) as modified by Saifi(1940).This non-alcohol material can be removed by washing the alcohol-distillatewith purified heptane. Both alcohol and the interfering non-alcohol material were shownto accumulate in potatoes held in nitrogen.     

Surface Attachment of Listeria monocytogenes Is Induced by Sublethal Concentrations of Alcohol at Low Temperatures

Sublethal concentrations of ethanol or isopropanol increased attachment of Listeria monocytogenes at 10, 20, or 30°C; no induction occurred at 37°C. The alcohol induction phenotype was retained in sigB and cesRK mutants; however, the degree of induction was affected. These results suggest that alcohol may contribute to the persistence of L. monocytogenes.     

包埋-脱水法常温和低温保存绿色巴夫藻

用包埋-脱水法在常温和低温下保存绿色巴夫藻(Pavlova virdis),探讨了温度、光(暗)和含水量等因素对存活率的影响.结果表明,通过调节含水量、控制光(暗)条件以及使用甘油保护剂等措施,绿色巴夫藻在常温和低温下都可以保存6个月并保持较高的存活率.其中4℃(暗)保存的最高存活率高达77.6%,而且保存后的藻细胞经过适当的恢复培养后,其生长力可以达到保存前的水平.包埋-脱水法操作简单,无需贵重设备,在藻类种质保存中有广阔的应用前景.     

Regulation of Bud Rest in Tubers of Potato, Solanum tuberosum L: VI. Biochemical Changes Induced in Excised Potato Buds by Gibberellic Acid

The rest period of the potato tuber was studied in relation to certain biochemical changes that are induced by gibberellic acid (GA₃). The concentration of reducing sugars in excised plugs with buds treated with 10⁻⁴m GA₃ decreased in the first 4 hours after treatment and then rapidly increased up to 70 hours. The pattern in control buds was similar, but the changes occurred more slowly. The response to GA₃ is temperature-dependent and is not limited to any particular tissue of the tuber. The concentration of reducing sugars in excised buds increased proportionally to the log of the concentration of GA₃ in a range from 10⁻⁸ to 10⁻⁴m. At 10⁻³m, GA₃ slightly inhibited production of reducing sugars. Malonate inhibits the initial decrease and the subsequent increase in reducing sugars in control buds, but not the increase induced by GA₃.     

Effect of Storage in Darkness and in Light on the Content of Membrane Lipids of Potato Tubers

The lipid composition of tubers from the potato varieties Bintje and Desirée was investigated during storage. Storage in total dark ness after harvest gave only small changes in the amounts of triglycerides. monogalactosyl diglycerides and digalactosyl digtycerides. Storage in light resulted in changes in these lipids and in their fatty acid composition. The absolute amount and the relative content of linolenic acid in the galactolipids increased. A simultaneous and equivalent decrease in the percentage of linoleic acid took place without any marked percentage changes in the other major fatty acids. The light induced changes of the lipids, which occur simultaneously with greening of the tuber, are discussed and related to the development of thylakoid membrane systems in the plastids.     

Active RNA Silencing at Low Temperature Indicates Distinct Pathways for Antisense-Mediated Gene-Silencing in Potato

Previously, it was shown that low temperature (≤ 15 °C) inhibits RNA silencing-mediated defence by the control of siRNA generation. In contrast, we have found nine antisense potato lines out of 24 in which RNA silencing was not inhibited at low temperature. In these lines, the extent of endogenous repression varied in leaves and was found to be different in roots and tubers. In order to address the contribution of gene dosage and repetitive structure of the transgene loci to the temperature dependence/independence of antisense-mediated gene-silencing DNA gel blot analysis was performed. Interestingly, none of the studied features correlated with the observed silencing effect. In addition, the insertion of vector backbone sequences into the potato genome did not influence the temperature dependence. RNA-directed DNA methylation was detected in the majority of antisense lines, however, it was also independent of the type of RNA silencing. Thus, it is feasible that chromosomal flanking sequences or the chromatin structure surrounding the insertion determine which silencing pathway is activated.     

Changes in Glycolytic Activity of Lactococcus lactis Induced by Low Temperature

The effects of low-temperature stress on the glycolytic activity of the lactic acid bacterium Lactococcus lactis were studied. The maximal glycolytic activity measured at 30°C increased approximately 2.5-fold following a shift from 30 to 10°C for 4 h in a process that required protein synthesis. Analysis of cold adaptation of strains with genes involved in sugar metabolism disrupted showed that both the phosphoenolpyruvate-dependent sugar phosphotransferase system (PTS) subunit HPr and catabolite control protein A (CcpA) are involved in the increased acidification at low temperatures. In contrast, a strain with the PTS subunit enzyme I disrupted showed increased acidification similar to that in the wild-type strain. This indicates that the PTS is not involved in this response whereas the regulatory function of 46-seryl phosphorylated HPr [HPr(Ser-P)] probably is involved. Protein analysis showed that the production of both HPr and CcpA was induced severalfold (up to two- to threefold) upon exposure to low temperatures. The las operon, which is subject to catabolite activation by the CcpA-HPr(Ser-P) complex, was not induced upon cold shock, and no increased lactate dehydrogenase (LDH) activity was observed. Similarly, the rate-limiting enzyme of the glycolytic pathway under starvation conditions, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), was not induced upon cold shock. This indicates that a factor other than LDH or GAPDH is rate determining for the increased glycolytic activity upon exposure to low temperatures. Based on their cold induction and involvement in cold adaptation of glycolysis, it is proposed that the CcpA-HPr(Ser-P) control circuit regulates this factor(s) and hence couples catabolite repression and cold shock response in a functional and mechanistic way.     

Biphasic Stimulation of Translational Activity Correlates with Induction of Translation Elongation Factor 1 Subunit [alpha] upon Wounding in Potato Tubers

Potato (Solanum tuberosum) tubers exhibit an increase in translational activity in response to mechanical wounding. The response is biphasic, with an initial stimulation apparent within the first 2 h after wounding and a second increase occurring 12 to 24 h after wounding. Increased activity is apparent by measurement of protein synthesis both in vivo and in vitro using a cell-free extract. Accumulation of the translational elongation factor 1 subunit [alpha] (EF-1[alpha]) parallels translational activity. Changes in the steady-state level of EF-1[alpha] mRNA, and expression of a chimeric EF-1[alpha] promoter/[beta]-glucuronidase construct in transgenic potato tubers, indicate that the gene encoding EF-1[alpha] is transcribed during both periods of translational stimulation. These results indicate that stimulation of translational activity is coordinated with increased expression and accumulation of translation factors.     

Studies on Endogenous Bacteria in Potato Tubers Infected by Phytophthora erythroseptica Pethybr

Infection of potato tubers by the soft-rotting fungi Phytophthoraerythroseptica and Pythium debaryanum resulted in multiplicationof endogenous tuber bacteria. This effect was not evident afterinfection by dryrotting fungi. Some of the bacteria isolatedproduced pectolytic and hemicellulolytic enzymes in vitro andwere themselves capable under certain conditions of degradingtuber tissue.     

Stress-Induced Translational Control in Potato Tubers May Be Mediated by Polysome-Associated Proteins

Potato tubers exhibit distinct responses to wounding and hypoxia that include selective translation of stress-induced mRNAs. Newly synthesized wound-response mRNAs are bound to polysomes, whereas preexisting mRNAs are displaced and degraded. mRNAs that are induced and translated during hypoxic conditions are bound to ribosomes as expected. However, preexisting wound-response mRNAs whose translation is inhibited during hypoxia remain bound to polysomes, indicating that there are at least two distinct mechanisms by which translation is regulated in response to stress conditions. A 32-kD phosphoprotein is associated with polyribosomes from wounded tubers. This protein remains polysome bound as long as wound-response mRNAs are present, even during hypoxia when these mRNAs are no longer translated. However, association of the 32-kD protein with polysomes is not elicited by hypoxic stress alone. The kinase that phosphorylates this protein is active only for the first 24 hr after wounding and is not active during periods of hypoxia. This protein may mediate recognition of the wound-response mRNAs by ribosomes.     

Light-Induced Chloroplast [alpha]-Amylase in Pearl Millet (Pennisetum americanum)

In pearl millet (Pennisetum americanum) seedlings light induces the appearance of a leaf [alpha]-amylase isozyme. The leaf [alpha]-amylase isozyme was present in enriched amounts in isolated chloroplast but it could not be detected in isolated etioplasts. The chloroplast [alpha]-amylase was present in both mesophyll and bundle-sheath chloroplasts. Preliminary characterization indicated that molecular properties of chloroplast [alpha]-amylase were like those of a typical [alpha]-amylase. The plastidic [alpha]-amylase had a molecular mass of 46 kD, pH optimum of 6.2, required Ca2+ for activity and thermostability, but lost activity in the presence of ethylenediaminetetracetate. Plastidic [alpha]-amylase activity after sodium dodecyl sulfate-polyacrylamide gel electrophoresis could be renatured in situ by Triton X-100. Western blot analysis demonstrated that this protein was antigenically similar to a maize seed [alpha]-amylase. In vivo [35S]methionine labeling of bundle-sheath strands isolated from light-grown leaves followed by immunoprecipitation revealed that bundlesheath strands synthesized plastidic [alpha]-amylase de novo.     

A Note on the Presence of {beta}-Amylase in Moulds

Evidence is given for the presence, in the mould Mucor Rouxianusand in several Rhisopus species, of an amylolytic enzyme producingglucose directly from starch. This glucosidase probably possessesa wide specificity range and attacks maltose as well. It doesnot destroy the iodine reaction of starch, but as a rule itis accompanied by another amylase which does destroy thb reaction.Of the mould amylases investigated, Mucor-amylase containedthe highest proportion of the glucosidase.     

Death of Cerebellar Granule Neurons Induced by Piperine Is Distinct from that Induced by Low Potassium Medium

We compared neurotoxicity of piperine and low K⁺on cultured cerebellar granule neurons. As considered from lactate dehydrogenase release and 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide reduction, both piperine and shifting from high K⁺(25 mM) to low K⁺(5.4 mM) were cytotoxic to cerebellar granule neurons. Protein synthesis inhibitors, cycloheximide and anisomycin, and an endonuclease inhibitor, aurintricarboxylic acid, were protective against low K⁺-induced neuronal death whereas they were ineffective against that induced by piperine. D--tocopherol, trolox, and a spin trap 3,3,5,5-tetramethyl-l-pyrroline-l-oxide were protective against piperine neurotoxicity whereas they had no effect on that induced by low K⁺. These results suggest that piperine and low K⁺may exert neurotoxic effects on cerebellar granule neurons through different mechanisms. Death of cerebellar granule neurons induced by piperine may be mediated by non-apoptotic mechanisms and may involve membrane lipid peroxidation and/or free radical generation.