Nature of enhanced respiration during sprouting of aged potato seed-tubers

Respiration of 18-month-old Solarium tuberosum L. tubers was about 53% greater than that of 6-month-old tubers during sprouting at 23°C; yet, a significant loss of sprout vigor in the older tubers was apparent. Involvement of alternative oxidase (AO) in the age-induced difference in tuber respiration was assessed. AO was only detected in immunoblots if tissue disks from tubers were pre-incubated for 24 h prior to isolation of submitochondrial membrane particles (SMPs). No AO¹ was detected in SMPs from nonincubated tuber tissue of either age, indicating that it was not contributing to tuber respiration during sprouting as previously thought. Respiratory control and ADP/O ratios indicated that oxidative phosphorylation was fully coupled to electron transport in mitochondria isolated from 6- and 18-month-old tubers. Cytochrome c oxidase (EC 1.9.3.1) activities of intact mitochondria were also not affected by tuber age. The difference in respiration during sprouting was unique to whole tubers, as oxygen consumption by mitochondria from young and oid tubers was equal on a milligram protein basis. Sprouting 18-month-old tubers had 15% more mitochondrial protein per gram fresh weight than did 6-month-old tubers. Older tubers also produced more ATP than younger tubers prior to and during sprouting, through a fully coupled, Cyt-mediated respiratory pathway, reduced sprout vigor notwithstanding. From 5 to 10 days of sprouting, coinciding with development of the age-induced difference in whole-tuber respiration, ATP concentration in 18-month-old tubers increased to become 52% higher than that in 6-month-old tubers. ATP synthase (EC 3.6.1.34), assessed by SDS-PAGE and immunoblots of β- and oligomycin-sensitivity conferring protein-subunits, also increased as a proportion of SMP protein in older tubers during this period. Relative to 6-month-old tubers, the increased respiration and associated oxidative phosphorylation of 18-rnonth-old tubers during sprouting were probably in response to a lower adenylate energy charge (AEC) prior to sprouting (from 0 fo 5 days). From 5 to 10 days of sprouting, AEC of 18-rnonth-old tubers increased to equal that of 6-month-old tubers and the two tuber ages maintained the same AEC for the remainder of the 20-day sprouting interval. Higher respiration and lower AEC of older tubers in storage at 4°C, along with the fact that older tubers respired at a higher rate to achieve the same AEC as younger tubers during sprouting, indicate greater utilization of ATP by older tubers.     

Deoxyuridine triphosphatase expression defines the transition from dormant to sprouting potato tuber buds

Identification of molecular markers defining the end of tuber dormancy prior to visible sprouting is of agronomic interest for potato growers and the potato processing industry. In potato tubers, breakage of dormancy is associated with the reactivation of meristem function. In dormant meristems, cells are arrested in the G₁/G₀ phase of the cell cycle and re-entry into the G₁ phase followed by DNA replication during the S phase enables bud outgrowth. Deoxyuridine triphosphatase (dUTPase) is essential for DNA replication and was therefore tested as a potential marker for meristem reactivation in tuber buds. The corresponding cDNA clone was isolated from potato by PCR. The deduced amino acid sequence showed 94% similarity to the tomato homologue. By employing different potato cultivars, a positive correlation between dUTPase expression and onset of tuber sprouting could be confirmed. Moreover, gene expression analysis of tuber buds during storage time revealed an up-regulation of the dUTPase 1 week before visible sprouting occurred. Further analysis using an in vitro sprout assay supported the assumption that dUTPase is a good molecular marker to define the transition from dormant to active potato tuber meristems.     

Involvement of Ethylene in Potato Microtuber Dormancy

Potato (Solanum tuberosum L.) single-node explants undergoing in vitro tuberization produced detectable amounts of ethylene throughout tuber development, and the resulting microtubers were completely dormant (endodormant) for at least 12 to 15 weeks. The rate of ethylene production by tuberizing explants was highest during the initial 2 weeks of in vitro culture and declined thereafter. Continuous exposure of developing microtubers to the noncompetitive ethylene antagonist AgNO₃ via the culture medium resulted in a dose-dependent increase in precocious sprouting. The effect of AgNO₃ on the premature loss of microtuber endodormancy was observed after 3 weeks of culture. Similarly, continuous exposure of developing microtubers to the competitive ethylene antagonist 2,5-norbornadiene (NBD) at concentrations of 2 mL/L (gas phase) or greater also resulted in a dose-dependent increase in premature sprouting. Exogenous ethylene reversed this response and inhibited the precocious sprouting of NBD-treated microtubers. NBD treatment was effective only when it was begun within 7 d of the start of in vitro explant culture. These results indicate that endogenous ethylene is essential for the full expression of potato microtuber endodormancy, and that its involvement may be restricted to the initial period of endodormancy development.     

The inhibition of potato sprout growth by light.

When potato seed tubers (Solanum tuberosum cv. Pentland Javelin) were stored in darkness or diffuse daylight at 9°C and transferred at intervals to conditions suitable for sprouting, their capacity for sprout growth was unaffected by the presence or absence of light during previous storage. When similar tubers were stored at 10°C, 18°C or 25°C, sprout growth commenced earliest at 25°C, but the date was unaffected by fluorescent light. It was concluded that light did not affect the length of the dormant period, but only the rate of sprout elongation after that period had ceased. When tubers with growing sprouts at 10°C or 18°C were transferred from darkness into fluorescent light, sprout growth virtually ceased. Transfer from light into darkness resulted in immediate sprout growth, at a rate comparable with tubers stored continuously in the dark. Tubers of three Peruvian cultivars, stored in farm-scale diffuse-daylight stores, grew progressively shorter sprouts with increasing daily exposure to light from 30 min to 12 h. Storage of cv. Wilja under 21 Wm^-2 (total) of white fluorescent light for 10 h per day maintained the sprouts at the same length as ten times this light intensity for 1 h per day. In a subsequent experiment with cv. Bintje the 10 h, low-intensity light regime gave slightly shorter sprouts. It appeared that the total light energy falling on the tubers was the dominant factor controlling sprout growth.     

Age of potato seed-tubers influences protein synthesis during sprouting

The effect of seed-tuber age on the ability of tuber tissue to synthesize protein during sprouting was examined. As seed-tuber age advanced from 4 to 32 months (at 4°C, 95% relative humidity), soluble protein concentration of tubers decreased linearly, with a concomitant increase in free amino acid concentration. The age-induced loss of tuber protein may thus be due to increased proteolysis, decreased protein synthesis, or both. Five- and 17-month-old seed-tubers were compared for their ability to incorporate radiolabeled amino acids into soluble protein at equivalent stages of sprout development. Tuber respiration was profiled through each sprouting stage to characterize the physiological status of the seed-tubers prior to incorporation studies. Five-month-old seed-tubers maintained a constant rate of respiration during sprouting. In contrast, respiration of 17-month-old tubers increased as sprout dry matter increased, resulting in a 2- to 3-fold greater respiratory rate from the older tubers, relative to the younger tubers, at similar stages of sprout development. Prior to sprouting, the rate of incorporation of amino acids into trichloroacetic acid-precipitable protein of tissue from 5-month-old tubers was 2. 9-fold higher than that from 17-month-old tubers. More importantly, protein-synthetic capacity of tissue from younger tubers increased about 1. 7-fold during sprout development. Despite the higher respiratory activity and faster total sprout dry matter accumulation from older seed-tubers, protein synthesis remained at a low and constant level through all stages of sprouting. Protein-synthetic capacity thus declines with advancing tuber age, and this may contribute to reduced growth potential during the latter stages of establishment by affecting the ability of seed-tubers to synthesize enzymes involved in mobilization and translocation of tuber reserves to developing plants.     

Effect of vitamins on sprouting of purple nutsedge tubers

Vitamins of the B group and vitamin C were applied to purple nutsedge (Cyperus rotundus L.) tubers to study their effect on the sprouting rate, initiation and establishment of sprouts, growth of plantlets and development of orthotropic rhizomes in comparison with the corresponding effects of kinetin. Ascorbic acid up to 100 mg I^?1 hastened sprouting, whereas vitamins of the B group and kinetin retarded sprouting; 100% tubers sprouted in all treatments within 10 days. Unlike kinetin, none of the vitamins resulted in the establishment of more than one sprout per tuber. Riboflavin and pyridoxine promoted root and shoot growth of the plantlets, whereas kinetin produced short, thick shoots and inhibited root growth, with increasing concentration. Ascorbic acid was only second to kinetin in the induction of orthotropic rhizomes, but the former resulted in an increase in rhizome length.     

Heat treatments for the control of sprouting of potatoes during storage

Potatoes were given hot water dip and vapour heat treatments at temperatures ranging from 50 to 80°C and 60 to 70°C respectively for various durations to control sprouting. Hot water treatment at its effective temperatures and durations resulted in undesirable physical damage to tubers. On the other hand, vapour heat treatment at 60°C with 95 ° 5 r.h. for 60 min suppressed sprouting, when given to tubers at the sprout emergence stage, without causing any deleterious side effects. The treatment, when repeated after 3 wk further reduced sprout yield during storage at ambient conditions (22–35°C, 50–80 r.h.). Comparative studies on the efficacy of vapour heat treatment and manual de-sprouting showed that vapour heat was superior to manual de-sprouting.     

Coiled-sprout in the potato: the effect of various storage and planting conditions

The incidence of coiled-sprout was determined in Scotch and local-grown Arran Pilot and Duke of York seed tubers which had been stored at 10°, 4° and 15° C. and in a farm store with no temperature control. All four tuber types were planted in the field and, in addition, the two types of Duke of York were planted in Perlite at 7°, 10° and 15° C. In the field, and when maintained at 7° and 10° C, the percentage of sprouts coiling and the intensity of coiling was greater in tubers stored at 10° and 15° C. than at 4° C. There was no coiling when the Duke of York tubers were planted at 15° C. In a further experiment tubers were stored at 20° C. in the light and dark and samples were planted monthly for 3 months at temperatures of 7°, 10° and 15° C. During the following 3-month period only light-stored were planted because of the excessive amount of tip-death in the tubers stored in the dark. There was very little coiling in the dark-stored tubers. In the first two plantings of the light-stored tubers there was virtually no coiling of those planted at 15° C. There was some, however, at 7° and 10° C. In subsequent plantings there was more coiling and no effect of planting temperature. Attempts to isolate Verticiculum nubilum from sprouts were successful in only a small percentage of attempts and it was not possible to demonstrate any difference between its distribution on coiled and normal sprouts. It was not possible to induce coiling by infection of sprouts with spores of V. nubilum. Over a wide range of sprout sizes the amount of coiling was a function of sprout size at planting. However, the parts which coiled were those in the apical bud of the sprout at the time of planting and hence contributed only a small amount to the total sprout size. It is likely, therefore, that the correlation between coiling and sprout size reflects the changing metabolism of the elongating regions of sprouts with their increase in length, these regions developing in such a way as to produce a greater tendency to coiling. The internal reactions concerned in these changes, however, are not known.     

Genotypic influence on in vitro induction, dormancy length, advancing age and agronomical performance of potato microtubers (Solanum tuberosum L.)

Microtubers of 13 cultivars, largely grown in Italy and other European countries, were induced. They were stored in the dark at 3°C for different periods (28, 56, 84 and 105 days), prior to being transferred to 20°C for between 4 and 17 weeks. Following removal to room temperature, sprouting was recorded and dormancy duration quantified. Dormancy decreased from 28.1 to 19.9, 11.1 and 7.8 days with reduced time of storage. Cvs Arsy, Nicola and Jaerla took consistently more time for dormancy release. The dormancy duration was linearly and inversely correlated with the length of storage. After sprouting, tubers were held at 20°C for various intervals and a range of physiological ages (0, 368, 720 and 1008 degree days) were accumulated. The field comparison of microtubers evidenced a plant growth response and tuber yield/plant affected by the cultivar and physiological age. In early cultivars (Jaerla), a better performance was shown by younger tubers; the opposite trend was noted in Alpha (a later cultivar) with an increase in stems/plant, tubers/plant and tuber yield/plant for tubers with greater physiological age. Like conventional seed tubers, microtubers showed differences in optimum physiological age associated with cultivar earliness. This study has provided some indications on how to enhance emergence and haulm development of plants from microtubers.     

Sugar accumulation in chemically debudded potato tubers during cold storage

Potato tuber buds may be excised by immersion of the tubers in a mixture of EtOH-Me₂CO (1:1) for 4 hr. This enabled the study of the effect of tuber aging (at 17°) on the starch-to-sugar conversion during storage at 4°, in the absence of complications due to sprouting. Sugar accumulation during a two-week period of storage at 4° decreased with increasing time of prior storage at 17°.     

Control of potato tuber dormancy and sprouting by expression of sense and antisense genes of pyrophosphatase in potato

Inorganic pyrophosphate (PPi) is an enzyme involved in sugar metabolism in potato tubers. In our previous study, we isolated an inorganic pyrophosphatase (PPase) gene from potato and obtained the transgenic potato plants transformed with the sense and antisense PPase genes respectively. In the present experiment, the physiological indexes, tuber dormancy, and sprouting characteristics of the transgenic potatoes were analyzed and evaluated. The result showed that the PPase activity and the inorganic phosphate content of tubers were lower in the antisense transgenic plant lines but were higher in the sense transgenic plant lines, compared with wild-type tubers. Soluble sugars, such as glucose, fructose and sucrose increased in transgenic plants that had overexpression of the sense PPase gene, but decreased in the antisense transgenic plant lines, compared with wild-type tubers. Tuber sprouting time of the antisense transgenic plants were delayed for 2 and 3 weeks and reached the 100 % sprouting rate only after 14 and 16 weeks storage compared with the wild-type when tubers are stored under 25 and 4 °C, respectively. In contrast, tuber sprouting time of the sense transgenic plants was earlier by approximately 2 weeks than that of wild-type tubers under these storage temperatures.     

The inhibition of potato sprout growth by light.

The effect of light intensity on sprout growth in seed potato tubers (Solanum tuberosum) was examined using diffuse daylight in Peru and diffuse artificial light at Glasgow. Mean temperatures below 20 °C produced strong sprout growth that was inhibited by both daylight and artificial light, at visible irradiances above 0.01 Wm^-2. The percentage inhibition of sprout growth increased linearly with the logarithm of the irradiance, 50% inhibition being at 0.04 - 0.1 Wm^-2 provided that the temperature was suitable for substantial sprout growth in the absence of light. Cultivar and temperature had very little effect on the 50% inhibition point. At high irradiances growth inhibition was up to 95%, but the sprout length was never reduced to zero; short, robust green sprouts remained. Sprout numbers were increased by daylight, but not by artificial light. Diffuse daylight also reduced the total weight loss from seed tubers during a storage season of 180 days. At mean temperatures above 20 °C., sprout growth in the absence of light was much reduced and the effect of light on sprout elongation was less obvious.     

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     

In vitro micro-tuber initiation and dormancy in yam

Dormancy is a mechanism that regulates the timing of sprouting (germination) of affected plant parts as well as ensures that the food quality of edible parts is maintained in storage until the following growing season. In yam, however, little is known about the control of tuber initiation or tuber dormancy. The objective of this study was to determine the effects of selected plant growth regulators (PGRs) on tuber initiation and dormancy, using an in vitro system. In two replicated experiments, 2-chloroethylphosphonic acid (ethephon, an ethylene source), abscisic acid (ABA) and gibberellin (GA₃) – and their inhibitors silver nitrate, fluridone and 2-chloroethyl-trimethylammonium chloride, respectively – were added at two concentrations to the culture medium prior to explant culture. Dates of micro-tuber initiation and sprouting (end of dormancy) and tuber number were recorded. In the control (no PGR) in Experiment 1, micro-tubers were initiated at the base of the stem after 176 days and sprouted 235 days later, that is 411 days after culturing. Most PGR treatments had only small effects (±30 days) on the duration of dormancy and the time of micro-tuber initiation. However, in GA₃ micro-tuber initiation occurred after 76 days, about 100 days earlier than in the control, whereas fluridone affected the position of micro-tubers and duration of dormancy. With fluridone treatments, tubers were found at the base of the stem (normal position) and on lower and upper nodes. Lower node tubers sprouted within 225 days of culturing compared with about 420 days after culturing at other nodal positions and in other PGR treatments. These data suggest an important role for ABA and gibberellic acid in yam micro-tuber initiation and the induction of dormancy.     

Effect of a granulovirus on mortality and dispersal of potato tuber worm (Lepidoptera: Gelechiidae) in refrigerated storage warehouse conditions

Potato tuber worm (PTW), Phthorimaea operculella (Zeller), is a world-wide pest of potato. In rustic stores, PTW larvae can infest 100% of stored tubers. Treatment of tubers in rustic stores with the PTW granulovirus (PoGV) has been demonstrated to protect stored tubers. This is the first study to show the effects of PoGV for protection of tubers stored in refrigerated warehouse conditions. Tubers were treated by dipping in aqueous suspensions of PoGV or water. An estimated 0.0819 larval equivalents of virus or 1.88×10⁹ viral occlusion bodies were deposited on each kilogram of tubers. They were held at 16°C for 11 days before lowering the temperature by 0.5°C per day until 10°C was reached. The tubers were stored at this temperature for 53 days. Mean numbers of infested tubers at the end of the assay was affected by both pre-infestation rate and virus treatment. Mean numbers of infested tubers in the control treatment was 3 tubers per chamber higher than in the virus treatment providing strong evidence that PoGV controlled larvae and minimized spread into un-infested tubers. Of the larvae that were retrieved in virus-treated infested tubers, the mean mortality was 87% compared to 37% in controls.     

Elevated CO2 stimulates cells to divide in grass meristems: a differential effect in two natural populations of Dactylis glomerata

In this study, we tested the hypothesis that elevated [CO₂] shortens the cell cycle in meristems of Dactylis glomerata, more in a Portuguese population (38°53′N) than in a Swedish population (63°09′N). In the shoot meristem, the cell cycle shortened to about the same extent (～ 26%) in both populations exposed to the elevated [CO₂] treatment. In the root meristem, the cell cycle shortened by 17% in the Portuguese and by 8% in the Swedish population. However, the proportion of rapidly cycling cells increased in the Portuguese much more than in the Swedish population in both meristems. In the root meristem, there was a 1.86-fold increase in the Portuguese compared with a 1.31-fold increase in the Swedish. In the shoot meristem, the increases were 1.5–3-fold for the Portuguese and 1.2-fold for the Swedish. The data are consistent in showing that a major response to the elevated [CO₂] treatment was an increase in the proportion of cells that were cycling and that this was more marked for the Portuguese population. A more general response to the elevated [CO₂] treatment was a shortening of the cell cycle regardless of population.     

Temperature and Light Requirements for the Sprouting of Chilled and Unchilled Tubers of the Purple Nutsedge, Cyperus rotundus

Mature and immature tubers of purple nutsedge (Cyperus rotundus L.) chilled at 0°C in dry and wet conditions, were sprouted along with fresh, unchilled tubers over a range of temperatures (10°C-45°C) in light and darkness. Fresh immature tubers showed a high sprouting percentage at all temperatures between 20°C and 40°C, while the mature ones did so only at 30°C and 35°C. Chilling of dry tubers stimulated early sprouting and increased the maximum sprouting percentage of both the mature and immature tubers. Dry chilling also lowered the limit of favourable temperatures to 15°C in the case of mature tubers. Chilling of wet tubers had a depressing effect and no sprouting occurred below 30°C. At all temperatures, light apparently favoured the sprouting of both the mature and immature tubers (except mature wet-chilled ones at 35°C and 40°C). Immature tubers showed relatively higher sprouting percentage than the mature ones, both in light and darkness. Alteration of temperature requirements due to dry and wet chilling of the tubers is regarded as significant and functional in relation to the ecology of the species.