Propagation and tuberization of potato bud clusters from bioreactor culture

Summary Single node stem segments fromin vitro potato shoots cultured in liquid medium in the presence of ancymidol (23.4 μM) developed into bud clusters in either shaken flasks or bioreactor cultures. Buds on the clusters developed tubers after subculture to a tuber induction medium with 23.2 μM kinetin, 19.5 μM ancymidol, and 6-8% sucrose. The number of tubers per cluster and their size were higher in agar induction medium on top of which a second layer of liquid medium was added, than in liquid shake or bioreactor cultures. The highest increase in tuber size (i.e., 720 mg fresh weight after 7 weeks), was obtained in agar cultures flushed twice with liquid tuber induction medium. The potential of bioreactor cultures for potato bud proliferation and enhanced tuber development in double layer agar-liquid cultures is discussed.     

The control of bud dormancy in potato tubers

Potato (Solanum tuberosum L.) tuber buds normally remain dormant through the growing season until several weeks after harvest. In the cultivar Majestic, this innate dormancy persisted for 9 to 12 weeks in storage at 10° C, but only 3 to 4 weeks when the tubers were stored at 2° C. At certain stages, supplying cytokinins to tubers with innately dormant buds induced sprout growth within 2 d. The growth rate was comparable to that of buds whose innate dormancy had been lost naturally. Cytokinin-treatment did not accelerate the rates of cell division and cell expansion in buds whose innate dormancy had already broken naturally. Gibberellic acid did not induce sprout growth in buds with innate dormancy. We conclude that cytokinins may well be the primary factor in the switch from innate dormancy to the non-dormant state in potato tuber buds, but probably do not control the subsequent sprout growth.Abbreviations tio ⁶ade 6-(4-hydroxy-3-methylbut-trans-2-enyl amino)purine, zeatin - tio⁶ado 6-(4-hydroxy-3-methylbut-trans-2-enyl amino)-9--D-ribofuranosyl purine, zeatin riboside     

Relationships between temperature, development and survival of different life stages of the tomato fruit fly, Neoceratitis cyanescens

The development and survival of female Neoceratitis cyanescens (Bezzi) (Diptera: Tephritidae) from egg to complete ovarian maturation were studied in the laboratory at five different constant temperatures: 15, 20, 25, 30, and 35 °C. The aim of this study was to get information on the influence of temperature on pre-mature stages, as a prerequisite to optimise rearing procedures and to understand temporal and geographical patterns of fruit fly occurrence. The developmental rate of the different life stages increased linearly with increasing temperatures up to 30 °C. The fastest development of pre-mature stages was recorded at 30 °C (22±1 days) and the slowest at 15 °C (98±3 days). The day-degrees requirements (K) to complete total development were 432.6 day-degrees. Lower temperature thresholds were 11.4, 11.9, 10.0, and 11.1 °C for egg, larval, pupal stages and ovarian maturation, respectively. The number of adults obtained from an initial batch of 100 eggs reached a maximum (64) at 25 °C. At 35 °C, no adults emerged. Larval developmental time was significantly shorter in green tomato fruits than in potato tubers at 15, 20, and 25 °C. Mortality rate of larvae was higher in green tomato fruits than in potato tubers at 25 and 30 °C.     

Influence of explant,temperature and different culture vessels on <Emphasis Type="Italic">in vitro </Emphasis>culture for germplasm maintenance of four mint accessions

Two experiments were conducted with four accessions of mint (Mentha spp.) on MS medium for their in vitro performance. In the first experiment apical and nodal explants were cultured at both 20 °C and 25 °C. Data was recorded at second and at fifth week. Both apical and nodal explants of mint showed better leaf production at 25 °C than 20 °C. Nodal explants of mint cultured at 25 °C in both cultivation periods exhibited the highest number of leaves. In the second experiment apical explants were cultured in four different culture vessels viz., industrial glass jar (IG), magenta vessel (MV), Erlenmeyer flask (EF) and culture tube with 1(CT1) and 2(CT2) explants at 25 °C for 6 weeks. The highest weight loss from the media, evapo-transpiration and fresh weight gain were recorded in IG and next in MV. The lowest weight loss from the media and fresh weight gain both were found in CT2. However the lowest evapo-transpiration was noted in EF. The highest numbers of leaves were recorded from MV. Without explants, depletion of medium and increase of headspace were both higher in IG than in the other vessels. Overall, Magenta vessel GA 7 showed the best in vitro performance.     

Response of potato grown under non-inductive condition paclobutrazol: shoot growth, chlorophyll content, net photosynthesis, assimilate partitioning, tuber yield, quality, and dormancy

The effect of foliar and soil applied paclobutrazol on potato were examined under non-inductive condition in a greenhouse. Single stemmed plants of the cultivar BP1 were grown at 35(±2)/20(±2) °C day/night temperatures, relative humidity of 58%, and a 16 h photoperiod. Twenty-eight days after transplanting paclobutrazol was applied as a foliar spray or soil drench at rates of 0, 45.0, 67.5, and 90.0 mg active ingredient paclobutrazol per plant. Regardless of the method of application paclobutrazol increased chlorophyll a and b contents of the leaf tissue, delayed physiological maturity, and increased tuber fresh mass, dry matter content, specific gravity, dormancy period of the tubers. Paclobutrazol reduced the number of tubers per plant. A significant interaction between rates and methods of paclobutrazol application were observed with respect to plant height and tuber crude protein content. Foliar application gave a higher rate of net photosynthesis than the soil drench. Paclobutrazol significantly reduced total leaf area and increased assimilate partitioning to the tubers. The study clearly showed that paclobutrazol is effective to suppress excessive vegetative growth, favor assimilation to the tubers, increase tuber yield, improve tuber quality and extend tuber dormancy of potato grown in high temperatures and long photoperiods.     

The relation between light and temperature effects in the induction and release of dormancy in the aerial tuber of Begonia evansiana

The effect of light quality on the induction and release ofbud dormancy in aerial tubers of Begonia evansiana ANDR. andthe relation in controlling the mechanism of dormancy betweenlight and temperature were investigated using aerial tubersin different dormant states; immature tubers which were sproutedby irradiation and mature ones which were sprouted only afterchilling. Blue or far-red light induced photo-sprouting in immature tubersat higher temperatures and also promoted release of dormancyin mature tubers caused by chilling (2–5°C). Blueor far-red light, however, failed to break mature tuber dormancyat room temperature. The induction of dormancy in immature tubers was advanced atlow temperatures (15–17°C) which were inoperativefor photo-sprouting. Lower temperatures showed conflicting dualeffects of inducing and breaking dormancy. Red light inhibitednot only photo-sprouting in immature tubers but also dormancyrelease in mature tubers at low temperatures. These light- and temperature-dependent reactions are independentlyinvolved in dormancy regulation, the former is overcome by thelatter with dormancy development. (Received December 3, 1968; )     

Temperature and the biology and predation of Ilione albiseta (Diptera: Sciomyzidae) — Potential biological control agent of liver fluke

The effect of temperature on predation by Ilione albiseta (Diptera: Sciomyzidae) on Lymnaea peregra was investigated at 14°, 17°, 20°, 23° and 26°C. The mean dry weight of snail tissue (Lymnaea peregra) attacked and consumed per day by first and second instar I. albiseta larvae was highest at 20°C while for third instar and total larval duration period it was greatest at 23°C. The mean number of snails killed per day during the third instar was also highest at 23°C. The total amount of snail tissue consumed by I. albiseta larvae increased significantly from first to second instar and from second to third instar at each constant temperature. Mean survival period of unfed first instar larvae decreased from 28.4 days at 14°C to 11 days at 26°C and the mean length of the second instar cephalopharyngeal skeleton decreased with increasing temperatures. As temperature increased the rate of consumption of oxygen (dissolved in water) by first and third instar larvae rose.     

Effect of temperature on development, growth and feeding of Coccinella septempunctata and Hippodamia convergens reared on the tobacco aphid, Myzus persicae nicotianae

Preimaginal development, mortality, aphid consumption rate, and size and weight upon reaching the adult stage of the aphidophagous coccinellids Hippodamia convergens Guérin-Méneville and Coccinella septempunctata L. collected from Karditsa, central Greece, were examined at four constant temperatures (14, 17, 20 and 23 °C) and L16:D8. The coccinellids fed on the tobacco aphid, Myzus persicae nicotianae Blackman. Egg, larval and pupal mortality was highest at 14 °C reaching 85.0, 73.8 and 29.4% in H. convergens and 49.3, 75.4 and 58.8% in C. septempunctata, respectively. Total preimaginal development ranged from 57.2 to 70.4 days at 14 °C, and to 16.9 and 22.1 days at 23 °C in H. convergens and C. septempunctata, respectively. Heavier and larger adults of H. convergens were obtained at 17 and 20 ° C. In C. septempunctata temperature did not affect adult weight while the lowest size was observed at 14 and 17 °C. Day-degrees requirements for preimaginal development in H. corvengens were 212.9 above a developmental threshold of 11.0 °C. The corresponding values for C. septempunctata were 281.5 and 10.7 °C. In H. convergens total and daily aphid consumption ranged from 46.8 aphids at 14 °C to 85.0 aphids at 23 °C and from 1.5 aphids at 14 °C to 9.2 aphids at 23 °C, respectively. The corresponding values for C. septempunctata were 112.0 and 2.7 at 14 °C and 157.7 and 12.4 at 23 °C. The results show the high potential of both predators as biological control agents against the tobacco aphid. The knowledge obtained could be essential for their appropriate use and for the improvement of mass rearing systems.     

Effect of soil temperature on nutrient allocation and mycorrhizas in Scots pine seedlings

We studied the effect of soil temperature on nutrient allocation and mycorrhizal development in seedlings of Scots pine (Pinus sylvestris L.) during the first 9 weeks of the growing season. One-year-old seedlings were grown in Carex-peat from a drained and forested peatland at soil temperatures of 5, 9, 13 and 17 °C under controlled environmental conditions. Fourteen seedlings from each temperature treatment were harvested at intervals of three weeks and the current and previous year's parts of the roots, stems and needles were separated. Mineral nutrient and Al contents in all plant parts were determined and the tips and mycorrhizas of the new roots were counted. Microbial biomass C and N in the growth medium were determined at the end of the experiment. None of the elements studied, except Fe, were taken up from the soil by the seedlings during the first three weeks. Thereafter, the contents of all the elements increased at all soil temperatures except 5 °C. Element concentrations in needles, stems and roots increased with soil temperature. Higher soil temperature greatly increased the number of root tips and mycorrhizas, and the numbers of mycorrhizas increased more than did the length of new roots. Cenococcum geophilum was relatively more abundant at lower soil temperatures (5 and 9 °C) than at higher ones (13 and 17 °C). A trend was observed for decreased microbial biomass C and N in the peat soil at higher soil temperatures at the end of the experiment.     

Effects of storage conditions on sprouting of microtubers of yam (Dioscorea cayenensis–D. rotundata complex)

The control of field tuber dormancy in the yam (Dioscorea cayenensis–D. rotundata complex) is poorly understood. Although studies have examined single environmental factors and chemical treatments that might prolong tuber dormancy and storage, only a few were focused on further tuber sprouting. The present study concerns microtubers obtained by in vitro culture. When microtubers were harvested (after 9 months of culture) and directly transferred on a new medium without hormones, the tubers rapidly sprouted in in vitro conditions. No dormancy was observed in this case. Harvested microtubers were also stored dry in jars in sterile conditions during 2 to 18 weeks before in vitro sprouting. In this case, microtubers stored during 18 weeks sprouted more rapidly than those stored 8 weeks. A constant “dormancy-like period” (storage duration + sprouting delay) was observed, between 20 and 28 weeks respectively for the more rapid and the slower microtubers. The size of the tubers used for the storage had great influence on further sprouting. The larger they were, the better they sprouted. Light during storage had no effect on the sprouting delay while a temperature of 25 °C permit a quicker sprouting than 18 °C. The medium used to obtain microtubers could also have an effect on sprouting rate.     

Effects of Oxygen and Respiratory Inhibitors on Induction and Release of Dormancy in Aerial Tubers of Begonia evansiana

Effects of O₂ and some respiratory inhibitors on the induction and release of bud dormancy were examined with the aerial tubers of different ages of Begonia evansiana Andr. Oxygen was needed not only for tuber sprouting but also during the chilling process at 2 to 5 C to break tuber dormancy. If the mature tubers were exposed to blue light during the chilling period, their dormancy was strikingly released even by the chilling given under an O₂ concentration as low as 3%. Blue light pretreatment promoted photo-sprouting of immature tubers only when given under lower O₂ concentrations. On the other hand, red light became effective in inducing dormancy in the immature tubers and in prolonging dormancy in the mature tubers as O₂ tension was increased. This was also the case with the induction of dormancy in the immature tubers by exposing them to a lower temperature (17 C) in the dark. The development of dormancy was suppressed by 2,4-dinitrophenol, p-nitrophenol, and sodium azide.     

Stimulation of potato (Solanum tuberosum L.) tuberization by semicontinuous liquid medium surface level control

A procedure for the mass propagation of potato (Solanum tuberosum L.) tubers using a laboratory scale jar fermentor is described. Tuberization was clearly suppressed when the shoots were completely submerged in the liquid medium. Tubers were mainly formed at the medium surface. When shoots were cultured by the semi-continuous medium surface level control method, in which the medium surface level was raised or lowered throughout the culture period, tubers were induced and developed in every area in the jar fermentor. Tubers propagated by this method contained about 18 % (w/w) dry matter, slightly less than in field grown tubers, but most of the tubers weighing more than 0.2 g(FW) sprouted under room condition, without any acclimatization, during 3 month after the culture.     

The control of bud dormancy in potato tubers. Measurement of the seasonal pattern of changing concentrations of zeatin-cytokinins

A radioimmunoassay, combined with high-performance liquid chromatography, has been used to analyse the zeatin-type cytokinins of potato (Solanum tuberosum L. cv. Majestic) tubers and tuber buds throughout growth and storage. During tuber growth, zeatin riboside was the predominant cytokinin detected in all tissues. Immediately after harvest, the total cytokinin concentration fell dramatically in the storage tissue, largely as a consequence of the disappearance of zeatin riboside. During storage, levels of cytokinins in the storage tissue remained relatively constant, but increased in the tuber buds. In the buds of tubers stored at 2°C there was a 20-to 50-fold increase in total cytokinin over six weeks, coinciding with the natural break of innate dormancy. At 10°C the rise in the level of bud cytokinins was slower, correlating with the longer duration of innate dormancy. Injecting unlabelled cytokinins into tubers in amounts known to induce sprouting gave rise to increases in cytokinin concentrations in the buds of the same order as the increase associated with the natural break of dormancy. Metabolism of injected cytokinins was greater in non-dormant than in dormant tubers. The roles of cytokinin concentration and the sensitivity of the buds to cytokinin in the control of dormancy are discussed.Abbreviations CK cytokinin - FW fresh weight - HPLC high-performance liquid chromatography - RIA radioimmunoassay - tio⁶ade 6-(4-hydroxy-3-methylbut-trans-2-enylamino)-purine=zeatin - tio⁶adeglc⁹ 6-(4-hydroxy-3-methylbut-trans-2-enylamino)-9--D-glucopyranosyl purine=zeatin-9-glucoside - tio⁶ado 6-(4-hydroxy-3-methylbut-trans-2-enylamino)-9--D-ribofuranosyl purine=zeatin riboside - tio⁶ado-[³H]-diol a radioactive derivative of zeatin riboside, synthesised by periodate-oxidation followed by [³H]NaBH₄-reduction - tio⁶AMP 6-(4-hydroxy-3-methylbut-trans-2-enylamino)-9--D-5-phosphoribofuranosyl purine=zeatin riboside 5-monophosphate - t(ioglc⁴)⁶ade 6-(4-O--D-glucopyranosyl-3-methylbut-trans-2-enylamino)-purine=zeatin-O-glucoside     

Separate and interactive effects of competition and herbivory on the growth, expansion, and tuber formation of Hydrilla verticillata

This study examined the interaction and main-effect impacts of herbivory by the leaf-mining fly Hydrellia pakistanae and plant competition from Vallisneria americana on the growth, expansion and tuber formation of Hydrilla verticillata in a 2 × 2 factorial design experiment. The study was conducted in 14,000-L tanks, over two growing seasons. Each tank represented a single experimental unit and contained 32 1-L pots. At the beginning of the experiment half of these were planted with H. verticillata while the other half were either left empty or planted with V. americana (the competitor). H. pakistanae fly larvae (the herbivore) were added to tanks as appropriate. No significant interactions were identified between herbivory and competition on any parameter of H. verticillata growth analyzed (i.e., total tank biomass accumulation, total number of rooting stems, total tuber number, total tuber mass, and tuber size), indicating that the factors were operating independently and neither antagonism nor synergism was occurring. Both competition and herbivory impacted the growth of H. verticillata. H. verticillata plants grown in the presence of V. americana developed less total biomass, had fewer total basal stems, had fewer tubers and less tuber mass per tank, and produced significantly smaller tubers relative to control plants. Herbivory also suppressed H. verticillata biomass accumulation and tended to suppress the number and total mass of tubers produced in each tank. Both factors showed 30–40% reduction of total H. verticillata biomass, although the mechanism of impact was different. Competition suppressed expansion of H. verticillata into adjoining pots but had little impact on its growth in pots where it was originally planted. Herbivory resulted in a general suppression of growth of H. verticillata in all pots. Although herbivory significantly impacted H. verticillata biomass, it did not result in competitive release for V. americana under the current experimental conditions. We conclude that management activities that promote competition or herbivory will impact the growth and expansion of H. verticillata. Furthermore, since these factors operated independently, the combined use of both factors should be beneficial for suppression of H. verticillata dominance.     

Novel bioreactor technology for mass propagation of potato microtubers

Potato (Solanum tuberosum L. ssp. tuberosum) microtubers were produced in vitro with Liquid Lab™ Rocker system. A thin-layer liquid culture was applied together with a regular pitch in autoclavable simple plastic vessels. All cultures were carried out at room temperature without contamination problems. Each cultivar tested (Asterix, Timo, Van Gogh, Velox) formed microtubers in the Liquid Lab system. The mean number of microtubers per vessel (50 explants) varied between 30 (cv. Asterix in 8 weeks tuber induction) and 75 microtubers (cv. Velox in 11 weeks tuber induction). Majority (63%) of the microtubers was sufficient by size and weight (above 200 mg) for further storage at dormancy (4°C). The cv. Velox yielded the highest number of microtubers with cultivation capacity. As a result of prolonged microtuber induction of 2–3 weeks, more microtubers with competence for cultivation were obtained per cultivar, except for cv. Van Gogh. Still, the mean weight of Van Gogh microtubers was significantly higher after prolonged microtuber induction (0.67 g) than after short induction (0.51 g). In conclusion, Liquid Lab™ Rocker system is a novel, efficient and rapid system for mass propagation of potato.     

Tuber induction and initiation during production and early field growth of transplants from in vitro-derived potato plants

In transplants from in vitro‐derived plantlets from very early potato (Solanum tuberosum) cultivars, a lower degree of tuber induction at the time of field planting is thought to increase tuber production. Leaf‐bud cuttings were used to assess the progress to tuber induction in in vitro‐derived potato plantlets during the transplant production phase and after subsequent transplanting into the field. Induction and initiation of tubers on the same plants were assessed to study the effects of the duration of transplant production and conditions during transplant production for cv. Gloria (very early) and cv. Bintje (mid‐early). In vitro‐produced plantlets were not induced by the time of planting but rapidly progressed to the induced state thereafter. The progress in induction with time and the change in percentage of plants showing tubers fitted typical sigmoid curves. Plantlets achieved 50% induction ca 15 days after planting into in vivo conditions, and 50% tuber initiation usually occurred 10 days later. Shorter transplant production periods reduced the degree of induction of the transplants at field planting. Transplant production for more than 2 weeks was required to allow conditions during that period to affect induction or initiation. Long‐term non‐inducing conditions delayed the progress to tuber induction in cv. Gloria and delayed tuber initiation in both cultivars. Cv. Gloria showed no faster progress to induction than cv. Bintje but initiated tubers earlier. The results suggest that the relation between progress to induction and tuber initiation is cultivar dependent and that leaf‐bud cuttings can be used successfully in very young in vitro‐derived plants for assessing the progress to tuber induction.     

Changes in the levels of abscisic acid and its metabolites resulting from chilling of tomato fruits

The 6,6,6-[²H]-analogues of abscisic acid (ABA), phaseic (PA) and dihydrophaseic (DPA) acids were used in GC-MS-SIM determination of free and total alkali hydrolyzable ABA, PA and DPA in the pericarp of tomato (Lycopersicon esculentum L. cv. Pik Red) fruit. Determinations were made on breaker-stage fruit stored 1, 2, 3 or 4 weeks at 2.5°C or at 10°C, and after subsequent ripening for 1 week in darkness at 20°C. Two-fold increases in levels of ABA occurred after storage at low temperatures with a slightly but significantly greater increase in ABA level occurring with 2.5°C storage. These increases in ABA levels were not associated with the appearance of damage symptoms that occurred with storage at the chilling temperature (2.5°C). Differences in ABA metabolism were found resulting from storage at the two temperatures, 2.5 or 10°C. Significantly greater DPA levels were found after 10°C storage than after 2.5°C storage (2 weeks). Levels of ABA ester-conjugates increased with 20°C ripening only after 10°C storage while free ABA levels decreased after both storage temperature conditions. Levels of DPA conjugates also increased only after 20°C ripening following 10°C storage. A longer period of storage resulted in decreases of free DPA levels after 10°C storage but increased DPA levels were found after 2.5°C storage.Abbreviations ABA abscisic acid - PA phaseic acid - DPA dihydrophaseic acid - GC-MS-SIM gas chromatography-mass spectrometry-selected ion monitoring - HPLC high pressure liquid chromatography - fw. fresh weight author for correspondence     

Effects of photoperiod and raised winter temperatures on egg development and timing of oviposition in the willow psyllid Cacopsylla moscovita

Nymphal development of Cacopsylla moscovita (And.) (Homoptera: Psylloidea) takes place only on female catkins of Salix repens L. and close phenological synchrony is crucial because development times of catkins and nymphs are similar. Eggs are laid on catkins as soon as they develop and close synchrony between oviposition and budburst maximizes time available for nymphal development. Sampling adult C. moscovita in the field revealed little synchrony between egg development and budburst, with over 60% of females containing mature eggs four weeks before catkins first appeared. In the laboratory, egg development was influenced by both temperature and photoperiod. At 10°C, egg development occurred approximately one month earlier than at 5°C and two weeks earlier than in the field. Adult survival in the laboratory was substantially reduced at increased temperature, with only 20% of adults surviving longer than two weeks at 10°C, compared with over 95% at 5°C. Body condition (weight corrected for size) of males and females decreased significantly in the field over winter. However, body condition of females levelled off before budburst, coincident with egg development implying that females had resumed feeding. We discuss our results in relation to regulation of phenological synchrony between oviposition and catkin appearance.     

Potato tubers as a biofactory for recombinant antibodies

Potato tubers have been successfully used for high-level production of a recombinant single-chain Fv (ScFv) antibody. Ubiquitous high-level expression was achieved under control of the CaMV 35S promoter through retention of the scFv protein in the endoplasmic reticulum. Recombinant antibodies accumulated up to 2% of total soluble tuber protein. After 1.5 years of tuber storage at 4 °C still half of the amount of scFv present in freshly harvested tubers was detectable. Its specific activity did not decrease during tuber storage. Recombinant protein could be efficiently purified from crude extracts by affinity chromatography.     

Effect of Temperature on Longevity and Productivity in Drosophila Ananassae: Evidence for Adaptive Plasticity and Trade-Off Between Longevity and Productivity

The genetic response of body size to temperature in the laboratory provides an interesting example of phenotypic plasticity. We found that females of Drosophila ananassae reared to adulthood at 18°C showed significant increase in body weight as compared to females reared at 25°C. At a given temperature, early productivity and lifetime productivity were the highest when the rearing and test temperature were the same. The effect of test temperature was highly significant for total productivity and early productivity. The interaction between test temperature and development temperature was also highly significant. Effect of development temperature was not significant. The females reared at 18°C showed greater body weight but their productivity was not significantly higher than smaller females reared at 25°C. Thus, the usually close relationship between size and fecundity is lost when the size change is due to rearing temperature. These findings provide evidence for adaptive plasticity in D. ananassae. We also found a negative correlation (trade-off) between longevity and productivity, the first report of such a trade-off between longevity and productivity in D. ananassae.