Resistance of potatoes transgenic for a cry1Ac9 gene, to Phthorimaea operculella (Lepidoptera: Gelechiidae) over field seasons and between plant organs

Stable performance of insect‐resistant transgenic plants across field seasons and between plant organs damaged by the insect pest is critical for management of this resistance in the field. To evaluate this, potato (Solanum tuberosum) lines transgenic for a cry1Ac9 gene with resistance to potato tuber moth (Phthorimaea operculella) were established in the field during the southern hemisphere summers of 1997/98, 1998/99 and 1999/00 as small field plots, each of 10 plants. Replicate plots of the non‐transgenic parent cultivars (at least one for every three independently derived transgenic lines) were planted randomly throughout the trials. Field‐grown foliage was challenged with larvae in the laboratory and a growth index (GI) was calculated for recovered larvae from each transgenic and non‐transgenic potato line. Larval growth on young and mature leaves, and on newly harvested or stored tubers was also measured in the laboratory. Foliage from the transgenic lines inhibited larval growth in all seasons tested. For both control and transgenic lines, larvae had slightly lower GIs when reared on mature leaves compared with young leaves, although the correlation between mean GI for young and mature transgenic leaves was high (r = 0.97). The correlation between the mean GIs of larvae on newly harvested tubers and on those stored for 5 months was also high (r = 1.0). However, the GIs of larvae on newly harvested transgenic tubers were larger than on transgenic tubers stored for 5 months. The relative growth indices (RGI = mean GI/number days before final weighing) of larvae reared on newly harvested tubers from transgenic lines were generally higher than those from young transgenic foliage, while the RGIs of larvae reared on non‐transgenic tubers were slightly lower than those fed non‐transgenic foliage. The correlation between mean RGIs of larvae fed tubers or foliage was 0.62. The transgenic potato lines exhibited stable resistance to larvae across field seasons, between affected plant organs, and between plant organs of different ages.     

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.     

An integrated view of the hormonal regulation of tuber formation in potato (Solanum tuberosum)

The hormonal regulation of the consecutive steps in the formation of tubers on the potato plant ( Solanum tuberosum L.) is described and discussed. An integrated view of the complex regulation of the initiation and growth of stolons and tubers is presented, with special emphasis on the commonly observed lack of synchronization of the various steps in tuber formation within a plant.     

A transgenic study on affecting potato tuber yield by expressing the rice sucrose transporter genes OsSUT5Z and OsSUT2M

In many plants, sucrose transporters are essential for both sucrose exports from sources and imports into sinks, indicating a function in assimilate partitioning. To investigate whether sucrose transporters can improve the yield of starch plant, potato plants (Solanum tuberosum L. cv. Désirée) were transformed with cDNAs of the rice sucrose transporter genes OsSUT5Z and OsSUT2M under the control of a tuber-specific, class-I patatin promoter. Compared to the controls, the average fructose content of OsSUT5Z transgenic tubers significantly increased. However, the content of the sugars and starch in the OsSUT2M transgenic potato tubers showed no obvious difference. Correspondingly, the average tuber yield, average number of tubers per plant and average weight of single tuber showed no significant difference in OsSUT2M transgenic tubers with controls. In the OsSUT5Z transgenic lines, the average tuber yield per plant was 1.9-fold higher than the controls, and the average number of tubers per plant increased by more than 10 tubers on average, whereas the average weight of a single tuber did not increase significantly. These results suggested that the average number of tubers per plant showed more contribution than the average weight of a single tuber to the tuber yield per plant.     

Tuber Formation and Growth of in vitro Cultivated Transgenic Potato Plants Overproducing Phytochrome B

We studied the effect of the ectopic expression of the Arabidopsis PHYB gene, which encodes the phytochrome B (phyB) apoprotein, under the control of cauliflower mosaic virus 35S promoter on the photoperiodic response of tuberization and growth of potato (Solanum tuberosum L., cv. Désirée) transformed lines. Stem cuttings of transformed and control plants were cultured on Murashige and Skoog nutrient medium containing 5 or 8% sucrose in the phytotron chambers at 20°C under conditions of a long day (16 h), a short day (10 h), or in darkness. We showed that the overexpression of the PHYB gene enhanced the inhibitory effect of the long day on tuberization. In addition, tuber initiation in these transformed plants occurred at a higher sucrose concentration. The insertion of the PHYB gene decreased plant and tuber weights and shortened stems and internodes. Thus, we demonstrated the complex result of the PHYB gene insertion: it affected the photoperiodic response of tuberization, the control of tuber initiation by sucrose, and the growth of potato vegetative organs.     

Age-specific bioassays and fecundity of Phthorimaea operculella (Lepidoptera: Gelechiidae) reared on cry1Ac -transgenic potato plants

Variation in the susceptibility of lepidopterous pest larvae of different ages to transgenic crops and the potential for survivors to reproduce could have important consequences for the development of resistance in such pests. Experiments were undertaken in the laboratory to determine if larvae of the potato tuber moth, Phthorimaea operculella, of different ages (0 (< 1 day old), 3, 5, 7 days) varied in their susceptibility to cry1Ac9–transgenic potato (Solanum tuberosum) foliage grown in the glasshouse or field. The survival and fecundity of larvae reared on transgenic tubers was also determined in the laboratory. There were no apparent differences in susceptibility of larvae of different ages to transgenic foliage. Larvae fed glasshouse or field‐grown non‐transgenic foliage had significantly larger relative growth indices and more larvae pupated, than those fed transgenic foliage, regardless of larval age. Eggs from a laboratory colony were placed on transgenic or non‐transgenic tubers to measure survival and fecundity. Between 6% and 15% of eggs placed on transgenic tubers developed into pupae for three of the four transgenic potato lines tested. On one transgenic line, only six adults emerged from 1300 eggs. In contrast, between 71% and 97% of the eggs placed on non‐transgenic tubers developed into pupae. Male and female pupae from transgenic lines weighed less than those from non‐transgenic lines. The fecundity of females from two of four transgenic lines was lower than from the non‐transgenic parent cultivar. Although larvae of different ages did not exhibit any overall age‐dependent pattern of increasing or decreasing susceptibility to transgenic foliage of glasshouse or field‐grown plants, the ability of larvae to survive and reproduce on transgenic tubers suggests this pest has the ability to evolve resistance to the transgenic plants used in the present study.     

Field evaluation of transgenic potato plants expressing an antisense granule-bound starch synthase gene: increase of the antisense effect during tuber growth

Transgenic plants of a tetraploid potato cultivar were obtained in which the amylose content of tuber starch was reduced via antisense RNA-mediated inhibition of the expression of the gene encoding granule-bound starch synthase (GBSS). GBSS is one of the key enzymes in the biosynthesis of starch and catalyses the formation of amylose. The antisense GBSS genes, based on the full-length GBSS cDNA driven by the 35S CaMV promoter or the potato GBSS promoter, were introduced into the potato genome by Agrobacterium tumefaciens-mediated transformation. Expression of each of these genes resulted in the complete inhibition of GBSS gene expression, and thus in the production of amylose-free tuber starch, in mature field-grown plants originating from rooted in vitro plantlets of 4 out of 66 transgenic clones. Clones in which the GBSS gene expression was incompletely inhibited showed an increase of the extent of inhibition during tuber growth. This is likely to be due to the increase of starch granule size during tuber growth and the specific distribution pattern of starch components in granules of clones with reduced GBSS activity. Expression of the antisense GBSS gene from the GBSS promoter resulted in a higher stability of inhibition in tubers of field-grown plants as compared to expression from the 35S CaMV promoter. Field analysis of the transgenic clones indicated that inhibition of GBSS gene expression could be achieved without significantly affecting the starch and sugar content of transgenic tubers, the expression level of other genes involved in starch and tuber metabolism and agronomic characteristics such as yield and dry matter content.     

Interrelation Between Growth Rate of Individual Potato (Solanum tuberosum L.) Tubers and Their Cell Number

In potato plants fast and slow growing tubers develop on thesame plant. A hypothetical causality between tuber growth rateand tuber cell number was investigated by determining the tubercell number with the aid of an automatic counting procedure.Our data show a close correlation between tuber size and cellnumber over the whole range of tuber volumes considered (3–28cm³). If the influence of tuber size on cell number is eliminatedby means of a partial correlation analysis, the cell numberof the entire tuber is not significantly correlated with itsgrowth rate. An exclusive consideration of the smaller cells(10–30 µm) in the apical tuber region, where thecell division rate in potato tubers is highest, reveals a loosebut significant partial correlation to tuber growth rate (r= 0.383, P < 0.05). The growth rate of the slow growing tubers of any potato plantmay be enhanced by removing the fast growing tubers. In thefirst few days this enhanced growth rate is not due to a stimulationof cell division rate, but rather due to cell expansion. Potato, Solanum tuberosum L., tuber growth rate, tuber cell number     

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.     

Tuber production,dormancy and resistance against Phthorimaea operculella (Zeller) in wild potato species

The diversification of resistant potato varieties at a landscape level could slow adaptation by Phthorimaea operculella to potato resistance and promote sustainable crop protection. In this study, we assessed wild potato species as novel sources of foliage and tuber resistance against P. operculella. Tuber resistance was quantified for 136 and foliage resistance for 54 potato accessions representing 14 and nine potato species, respectively. Several accessions were highly resistant to moth damage in tubers and/or foliage. In particular, Solanum chiquidenum and Solanum sandemanii were highly resistant to damage in tubers. Several accessions of Solanum multiinterruptum and a small number of accessions of Solanum bukasovii, Solanum berthaultii, Solanum sparsipilum and Solanum wittmackii also had highly resistant tubers. Larval survival on foliage of S. bukasovii and S. chiquidenum was generally low. New resistance sources are listed, and insect performance on the plants is described with possible resistance mechanisms. The study also examined potential trade‐offs associated with resistance. Tuber resistance was negatively correlated with the number and weight of tubers produced per plant, but positively correlated with the length of dormancy across accessions, indicating that, although long dormancy is not a prerequisite for resistance, species and accessions with extended dormancy will have more resistant tubers. Tuber and foliage resistance were generally positively correlated across all accessions; however, among accessions from within a potato species, there were negative (S. berthaultii), positive (S. chiquidenum) and non‐significant (S. bukasovii) relations. These results indicate that, besides identifying novel resistance sources, an improved understanding of the mechanisms and inherent trade‐offs associated with tuber and foliage resistance will improve the efficiency of potato breeding programmes aimed at enhancing resistance against P. operculella.     

Theobroxide induces tubers in potato (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.) and flower buds in morning glory (<Emphasis Type="Italic">Pharbitis nil</Emphasis>) under non-inductive high temperatures

Induction of some plant organs including tubers and flower buds begins with sensing environmental cues, such as photoperiod and temperature in the leaves. Theobroxide has been shown to induce potato tuberization and flower-bud formation in morning glory under non-inductive photoperiodic conditions, stimulating the activity of lipoxygenase (LOX) and the synthesis of jasmonic acid (JA). In the present study, the ability of theobroxide to overcome the inhibitory effect of unfavorable high temperature on the induction of tubers in potato and flower buds in morning glory was examined. Both tuber induction and flower-bud formation under non-inductive high temperatures were promoted by the application of theobroxide at a high concentration. However, although theobroxide treatment resulted in an increase in fresh weight during potato tuber growth at 30°C, morning glory plants treated with theobroxide at 35°C failed to bloom, implying that theobroxide may assist only in flower-bud formation.     

Temporal dynamics of tuber formation and related processes in a crossing population of potato (Solanum tuberosum)

The chronological relationships between stolon formation, stolon tip swelling, tuber initiation, flowering, senescence, growth and resorption of tubers were studied under field conditions in a diploid population of potato with 238 genotypes, the parental clones and seven tetraploid cultivars. Timing of tuber initiation was not closely related to the timing of stolon formation, flowering and duration of the plant cycle. Tuber initiation very often preceded stolon branching. The number and size distribution of tubers were largely influenced by the degree of stolon branching, the length of the stolon swelling period and tuber resorption. The peak production of stolons and swollen stolon tips largely took place within the flowering period, although in most genotypes, some stolon tip swelling took place until the end of the plant cycle. More information on the general temporal relationships between events related to tuber formation and plant development will contribute to a better understanding of the physiological and genetic basis of the processes leading to the production of harvestable tubers.     

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.     

The three-dimensional distribution of minerals in potato tubers

Background and Aims

The three-dimensional distributions of mineral elements in potato tubers provide insight into their mechanisms of transport and deposition. Many of these minerals are essential to a healthy human diet, and characterizing their distribution within the potato tuber will guide the effective utilization of this staple foodstuff.

Methods

The variation in mineral composition within the tuber was determined in three dimensions, after determining the orientation of the harvested tuber in the soil. The freeze-dried tuber samples were analysed for minerals using inductively coupled plasma-mass spectrometry (ICP-MS). Minerals measured included those of nutritional significance to the plant and to human consumers, such as iron, zinc, copper, calcium, magnesium, manganese, phosphorus, potassium and sulphur.

Key Results

The concentrations of most minerals were higher in the skin than in the flesh of tubers. The potato skin contained about 17 % of total tuber zinc, 34 % of calcium and 55 % of iron. On a fresh weight basis, most minerals were higher in tuber flesh at the stem end than the bud end of the tuber. Potassium, however, displayed a gradient in the opposite direction. The concentrations of phosphorus, copper and calcium decreased from the periphery towards the centre of the tuber.

Conclusions

The distribution of minerals varies greatly within the potato tuber. Low concentrations of some minerals relative to those in leaves may be due to their low mobility in phloem, whereas high concentrations in the skin may reflect direct uptake from the soil across the periderm. In tuber flesh, different minerals show distinct patterns of distribution in the tuber, several being consistent with phloem unloading in the tuber and limited onward movement. These findings have implications both for understanding directed transport of minerals in plants to stem-derived storage organs and for the dietary implications of different food preparation methods for potato tubers.     

Growth and yield of potato plants inoculated with rhizosphere bacteria

Pot and field experiments on small plots were carried out to study the effect of isolates ofPseudomonas fluorescens-putida bacteria, applied to potato tubers, on the growth response of potato plants. Inoculation of tuber pieces with the isolates in pot experiments caused a better growth of young potato plant (111 % increase). The effect depended on the viable count in the bacterial suspension and on growth conditions of the plants. Inoculation of potato seed pieces before planting in field experiments caused a 4 –30 % improvement in plant growth and tuber yield. The complex action of inoculation with rhizobacteria apparently caused changes in the microflora colonizing the roots and stolons, which in turn brought about a better growth and yield.     

Translocation of Assimilates Within and Between Potato Stems

Three aspects of translocation in potato were examined: (i)translocation within stems (ii) translocation between individualstems of a plant (iii) translocation between tubers followinginjection of ¹⁴C sucrose into a single daughter tuber. Assimilatesexported from single leaves of evenly illuminated potato stemsremained confined to the same side of the stem as the sourceleaf in a pattern consistent with the internal arrangement ofvascular bundles in the stem, and tubers borne on stolons verticallybelow the source leaf contained higher concentrations of ¹⁴Cthan those on the opposite side. Consequently ¹⁴C import intothe tubers bore little relationship to tuber growth rates. However,alteration of source/sink relations by pruning stems to a singlesouce leaf resulted in an even distribution of ¹⁴C throughoutthe vascular bundles of the stem and ¹⁴C import into the tubersbore a stronger relationship to tuber growth rates than to thephyllotactic relationship of the tubers with the source leaf. Labelling one stem of a potato plant resulted in little or nomovement of ¹⁴C into tubers on other unlabelled stems. However,removal of the unlabelled stems at ground level induced a significantmovement of ¹⁴C from the labelled stem to the tubers on unlabelledstems, this movement occurring via the mother tuber. Shadingthe unlabelled stems had less effect than stem removal. ¹⁴C sucrose injected into single daughter tubers was translocatedto other tubers on the same stem and also to tubers on a secondstem at the opposite end of the mother tuber. The sucrose wasconverted to starch in these tubers. The results favour the view that each potato stem functionsas an independent unit with potential for assimilate redistributionwithin a stem but with little or no carbon exchange occurringbetween stems, unless under severely altered source/sink patterns. Assimilates, ¹⁴C, autoradiography, potato (Solanum tuberosum L.), tuber growth     

Correlation of cell division and cell expansion to potato microtuber growth in vitro

The sink capacity of plant storage organs influences crop economic yield and relates to the number and volume of their cells. To obtain a better understanding of their contributions to the growth of potato microtubers produced in vitro, the number and volume of the cells in the tuber tissues were measured as tubers grew. Two potato cultivars, E-Potato 1 and Mira were employed and the results showed that cortex, perimedulla and pith tissue contributed for about 30, over 65 and up to 3% to the volume of the mature microtuber, respectively. The number of cells and cell volume increased simultaneously as the microtubers grew and the relationships could be described by a power function, Y = aW ^b. However, the rate of cell division was greater than the rate of cell expansion and the former contributed more than the latter to the increase in tuber size. The rate of cell division was greatest in the cortex and least in the pith, but, because the perimedulla forms the largest part of the tuber, cell division in this tissue was particularly important. The regulation of cell division to improve the production of usable microtubers is discussed.     

StGA2ox1 is induced prior to stolon swelling and controls GA levels during potato tuber development

The formation and growth of a potato ( Solanum tuberosum ) tuber is a complex process regulated by different environmental signals and plant hormones. In particular, the action of gibberellins (GAs) has been implicated in different aspects of potato tuber formation. Here we report on the isolation and functional analysis of a potato GA 2-oxidase gene ( StGA2ox1 ) and its role in tuber formation. StGA2ox1 is upregulated during the early stages of potato tuber development prior to visible swelling and is predominantly expressed in the subapical region of the stolon and growing tuber. 35S-over-expression transformants exhibit a dwarf phenotype, reduced stolon growth and earlier in vitro tuberization. Transgenic plants with reduced expression levels of StGA2ox1 showed normal plant growth, an altered stolon swelling phenotype and delayed in vitro tuberization. Tubers of the StGA2ox1 suppression clones contain increased levels of GA₂₀, indicating altered GA metabolism. We propose a role for StGA2ox1 in early tuber initiation by modifying GA levels in the subapical stolon region at the onset of tuberization, thereby facilitating normal tuber development and growth.