Variations in resistance against Phthorimaea operculella in wild potato tubers

Tuber resistance can contribute to current management strategies against the potato tuber moth, Phthorimaea operculella Zeller (Lepidoptera: Gelechiidae), in field and stored potatoes. Wild potatoes represent a potential source of novel resistance traits against the moth. We assessed resistance in three wild potato species, Solanum multiinterruptum Bitt., Solanum sparsipilum (Bitt.) Juz. & Buk., and Solanum wittmackii Bitt. against neonate and developing tuber moth larvae. All three species had high levels of resistance but accessions of S. sparsipilum and S. wittmackii were significantly more resistant. Resistance in S. multiinterruptum was generally concentrated in the tuber periderm, whereas in S. sparsipilum and S. wittmackii resistance was mainly cortex‐based. Unidentified cortex‐resistance factors in all three species reduced survival and increased larval and pupal development times, but had no apparent effects on the pupal weights of survivors. A high proportion of larvae abandoned or died within tubers of S. wittmackii, which has particularly high levels of unidentified cortex‐based defenses. Resistance decreased in S. multiinterruptum and S. sparsipilum as tubers sprouted but was more stable in S. wittmackii. Periderm‐based resistance was more stable than cortex‐based resistance in S. multiinterruptum during sprouting. In contrast, cortex‐based resistance was stable in tubers of S. wittmackii as these sprouted, and resistance may have increased on some older sprouting tubers. Solanum multiinterruptum and S. sparsipilum are proposed as potential sources of resistance against the potato tuber moth.     

Interaction between day length and phytohormones in the control of potato tuberization in the in vitro culture

We studied the interaction of the day length, cytokinins, and gibberellins in the control of tuberization in potato (Solanum tuberosum L, cv. Desire) plants and derived transgenic plants with the inserted PHYB gene from Arabidopsis encoding the synthesis of phytochrome B apoprotein and put under the control of the 35S CaMV promoter. Plantlets were cultured in vitro on hormone-free MS medium containing 5% sucrose and kinetin (1 mg/l) or/and GA (0.5 and 1.0 mg/l), at long day (LD, a 16-h photoperiod), short day (SD, a 10-h photoperiod), or continuous darkness conditions. The content of cytokinins (Ck, zeatin, and zeatin riboside) in various plant organs was determined by the immunoenzyme method, and GA activity was measured in bioassay with dwarf pea. Potato plant transformation with the PHYB gene enhanced substantially tuber initiation inhibition by LD. Kinetin addition to culture medium enhanced tuberization and reduced Ck content in aboveground shoots and Ck redistribution in the favor of underground organs. GA addition to the culture medium suppressed tuberization and induced Ck accumulation in aboveground organs. We concluded that Ck role in tuberization depends on their predominant localization in above- or underground potato organs. The involvement of Ck and GA in the competitive relations between growing tubers and shoots is considered.     

The Interaction of Gibberellins and Photoperiod in the Control of Potato Tuberization

Solanum tuberosum ssp. andigena plants require a short-day (SD) photoperiod for tuber formation, a process that is also affected by gibberellins (GAs). Grafting experiments have confirmed that the photoperiod is perceived in the leaves. Tuber formation, however, usually takes place in the underground stolons. In this review, photoperiod-dependent tuberization has been divided into five chronological events: SD photoperiod perception, short-term adaptive responses to SD conditions, generation and transport of tuber-inducing signal(s), tuber formation, and long-term adaptive responses to tuber growth. Within this frame of study, the interaction of GAs and photoperiod is revised. Similar to the flowering process in Arabidopsis, we suggest the existence of two independent pathways that control tuber formation: a photoperiod-dependent pathway and a GA-dependent pathway. Nevertheless, photoperiod-dependent tuber formation requires the action of GAs at specific stages to orchestrate this complex process of development.     

Influence of Temperature and Photoperiod on Flowering and Tuberous Root Formation of Pachyrrhizus tuberosus

P. tuberosus, a native species of the Amazon region, was cultivatedunder different thermal regimes and photoperiods in an attemptto relate these stimuli to flower initiation and tuberous rootformation. It is shown that P. tuberosus is an intermediate-dayplant (flowers only under photoperiods above 9 h and below 16h). With regard to tuberization P. tuberosus may be considereda short-day plant: tuberization occurs only in photoperiodsbelow 16 h. High thermal regimes 30/25 C (13 h day/11 h night)delay and reduce flowering and completely inhibit tuberous rootformation. Thermal regimes of 25/20 C and 20/15 C (13 h day/11h night) were the most suitable for flowering and tuberization. Pachyrrhizus tuberosus, yam-bean, flowering, tuberization     

In vitro tuberization of potato clones from different maturity groups

Summary In vitro tuberization on shoot cultures of early, mid-season, late and very late potatoes was compared. Shoots were grown at 12, 16, or 20 h photoperiods; tuberization was then induced at 0, 8 or 16 h light. In the dark, shoots from early plants initially grown at 16 h consistently set tubers earlier than the other types, whereas the very late line tuberized later and produced significantly fewer tubers. Tuber setting of mid-season plants could not be distinguished from the late type. Tuberization of the very late line was significantly hastened by shortening the photoperiod from 20 h to 12 h during the shoot growth period. Light during tuber induction delayed tuberization. This system may be useful to screen callus-derived plants for maturity, and may also be suitable for in vitro study of the photoperiodic control of tuberization.     

QTL analysis of potato tuber dormancy

The potential loss of chemical sprout inhibitors because of public concern over the use of pesticides underscores the desirability of breeding for long dormancy of potato (Solanum tuberosum L.) tubers. Quantitative trait locus (QTL) analyses were performed in reciprocal backcrosses between S. tuberosum and S. berthaultii toward defining the complexity of dormancy. S. berthaultii is a wild Bolivian species characterized by a short-day requirement for tuberization, long tuber dormancy, and resistance to several insect pests. RFLP alleles segregating from the recurrent parents as well as from the interspecific hybrid were monitored in two segregating progenies. We detected QTLs on nine chromosomes that affected tuber dormancy, either alone or through epistatic interactions. Alleles from the wild parent promoted dormancy, with the largest effect at a QTL on chromosome 2. Long dormancy appeared to be recessive in the backcross to S. berthaultii (BCB). In BCB the additive effects of dormancy QTLs accounted for 48% of the measured phenotypic variance, and adding epistatic effects to the model explained only 4% more. In contrast, additive effects explained only 16% of the variance in the backcross to S. tuberosum (BCT), and an additional 24% was explained by the inclusion of epistatic effects. In BCB variation at all QTLs detected was associated with RFLP alleles segregating from the hybrid parent; in BCT all QTLs except for two found through epistasis were detected through RFLP alleles segregating from the recurrent parent. At least three dormancy QTLs mapped to markers previously found to be associated with tuberization in these crosses.Paper number 55 of the Department of Fruit and Vegetable Science, Cornell University     

Growth rate of potato tubers and endogenous contents of indolylacetic acid and abscisic acid

Potato plants ( Solanum tuberosum L. ev. Ostara) were grown in water culture and the growth rate of individual tubers was measured daily or at two day intervals. Tubers of different growth rate and/or different age (days after tuberization) were harvested and analysed for indolylacetic acid (IAA) and abscisic acid (ABA). Within individual tubers the IAA content decreases from the apical to the basal part of the tuber. Tuber age and corresponding fresh weight are negatively correlated with the endogenous IAA content. If, however, individual tubers of comparable age but different growth rates are compared, a significant positive correlation between growth rate and IAA content is revealed, while ABA showed a significant negative correlation with growth rate. Removal of all fast-growing tubers from individual plants causes an increase in the growth rate of the remaining tubers within 3–4 days. This coincides with a particularly steep increase in IAA content. The data support the idea that endogenous IAA content may be one factor responsible for controlling the growth rate ("sink-activity ") of individual tubers.     

Gibberellin A1 metabolism contributes to the control of photoperiod-mediated tuberization in potato

Some potato species require a short-day (SD) photoperiod for tuberization, a process that is negatively affected by gibberellins (GAs). Here we report the isolation of StGA3ox2, a gene encoding a GA 3-oxidase, whose expression is increased in the aerial parts and is repressed in the stolons after transfer of photoperiod-dependent potato plants to SD conditions. Over-expression of StGA3ox2 under control of constitutive or leaf-specific promoters results in taller plants which, in contrast to StGA20ox1 over-expressers previously reported, tuberize earlier under SD conditions than the controls. By contrast, StGA3ox2 tuber-specific over-expression results in non-elongated plants with slightly delayed tuber induction. Together, our experiments support that StGA3ox2 expression and gibberellin metabolism significantly contribute to the tuberization time in strictly photoperiod-dependent potato plants.     

Further Studies on the Relationship between Growth Regulators and Tuberization of Dahlias

Abscisic acid (ABA) and gibberellin (GA) treatments, applied to budless leaf cuttings of Dahlia under long day conditions, did not increase the total weight of the cuttings but did affect the region where photosynthates accumulate. GA treatment inhibited the growth of tubers and roots, and enhanced thickening at the petiole base. ABA did not affect the weight of roots or petiole, but promoted the growth of tuberous roots. Anatomical examination of the tuberization revealed that it is caused by inter-fascicular cambial activity producing parenchyma cells inward. No anatomical difference could be observed between the GA-induced thickening at the petiole base and the natural or ABA-induced thickening of the roots. In single node cuttings, taken from various sites along the branch, tuberization ability increased with proximity to the base of the branch, where axillary buds are more inhibited. Short-day conditions which promote tuberization, increased the endogenous level of ABA-like inhibitors in intact plants. External conditions and growth substances which inhibit growth also inhibited tuberization. Benzyladenine treatments did not affect tuberization. It is suggested that in addition to their respective indirect effects via the enhancement or suppression of top growth, GA and ABA directly control tuberization. ABA levels which increased under short-day conditions seem to produce a sink in the roots. GA probably inhibits tuberization by interfering with the trans-location of photosynthates to the root area.     

Phosphate transport in potato cuttings: Effect of gibberellic acid and abscisic acid

Apical cuttings of Solanum tuberosum L. cv. Sirtema were used al different stages of development to study long-distance transport of phosphate. The effects of two hormones, gibberellic acid (GA₃) and abscisic acid (ABA), on this process were also investigated. Before tuberization, phosphate (³²P) supplied to a single leaf was transported preferentially in the young and growing parts of the plant: apical bud, young leaves and roots. After tuberization, the tuber became the principal site of phosphate accumulation. GA³ treatment (10⁻⁴ M) of the tuber as well as of the leaves led to reduced transport of ³²P into the tuber. By contrast, treatment of the tuber with ABA (10⁻⁴M) did not change the ³²P distribution within the plant, while foliar spray with ABA greatly increased the transport into the tuber. The opposite effects of the two hormones on phosphate accumulation by tubers are discussed with regard to their opposite effects on the tuberization process.     

Endogenous inhibitors and seasonal changes in abscisic acid in Dioscorea floribunda Mart. & Gal.

In the tubers of medicinal yam (Dioscorea floribunda Mart. & Gal.) abscisic acid (ABA) content was high in all the parts during the dormant condition in the winter, but it decreased to a very low level in the actively growing plants. ABA content of the entire tuber was negatively correlated with temperature and photoperiod. Growth inhibitors including batatasin-I and phthalic acid were identified in the dormant tuber.CIMAP Communication No. 673.     

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.     

Photoperiodic control of growth, development and phytohormone balance in Solanum tuberosum

Some lines of Solanum tuberosum ssp. andigena are strictly photoperiodic, forming tubers only in short days and flowers only in long days. We used this advantageous phenomenon to study phytohormone involvement in the development of the plants, mainly that of tuber formation. Plants grown for 2 months under short days (SD) of 14 h darkness, night break (1 h white light in the middle of the 14‐h dark period) and continuous light (LD) were compared. Short day‐grown plants formed tubers, while plants in LD flowered. Night break prevented tuber formation, but caused flowering, although it was weaker than in LD. Plants grown under night break displayed many growth characteristics intermediate between SD and LD. Under LD and night break regimes ABA levels in all organs were about one‐fourth of those under SD. An opposite trend was found for gibberellin content: it was very low in SD‐grown plants and 4‐10 times higher under both other conditions with the exception of roots and stolons in night break. Thus, the ratio of ABA/GA, known to be important for tuber formation, was high in SD and low in night break and LD. The level of free indoleacetic acid (IAA) was high in LD‐ and night break‐grown plants and it was much lower in SD‐grown plants, including tubers. Cytokinin (CK) levels were also high in LD‐ and night break‐grown plants. In SD, leaves had slightly decreased and stems and roots + stolons had more pronouncedly decreased cytokinin levels. The CK/IAA ratio was thus higher in SD in stems and roots + stolons; it was also high in tubers and no difference was found in leaves. These results indicate that the effect of photoperiod on tuber formation and development may be mediated by photoperiod‐induced changes in hormone levels.     

Use of the growth retardant tetcyclacis for potato tuber formation in vitro

The effects of the plant growth retardant tetcyclacis on in vitro tuber formation in potatoes was studied, using two different approaches: 1. tuber formation in various lines that did not or hardly form tubers under control conditions, and 2. tuber formation by the variety Bintje, which readily forms tubers. The ABA-deficient (droopy) lines of S. phureja hardly formed tubers without the addition of tetcyclacis. In the presence of this growth retardant tuberization was nearly 100%, within three weeks of in vitro culture, even in the absence of cytokinin. A series of somatic hybrids between S. tuberosum and S. brevidens, that did not form tubers in field and pot experiments, were tested. They all formed tubers in vitro in the presence of tetcyclacis. Stoloniferous shoots formed on single-node cuttings from in vitro grown Solanum tuberosum var Bintje plantlets were transferred to media containing a high level of sucrose. In the presence of tetcyclacis, tuber formation started after 4 days, reaching a maximum level of 80% at day 7. Tubers formed in the presence of tetcyclacis, accumulated starch and expressed several tuber-specific genes. These effects were fully antagonized by gibberellic acid. It is concluded that the growth retardant tetcyclacis is a potent tool in the study of tuber formation in potatoes.Abbreviations ABA abscisic acid - BAP benzylaminopurine - GA₃ gibberellic acid - STS silver thiosulphate - TET tetcyclacis     

Flowering of Cultivated Green and SAN 9789-Treated Chenopodium rubrum Plants Exposed to White,Blue, and Red Light

Green plants and plants devoid of photosynthetic pigments were compared with regard to their ability to flower under various growth conditions. Green plants of Chenopodium rubrum L. and plants treated with norflurazon SANDOZ-9789 (SAN) were grown on sucrose-containing media with or without hormones (GA₃, BA, IAA, ABA) under short-day photoperiodic or continuous illumination with white, blue, or red light. Green and SAN-treated albino plants produced flowers only under short-day conditions. The flowering of green plants was independent of the presence of sucrose and hormones in the medium as well as of the light quality. The albino plants produced flowers under white and blue light but did not flower in red light. The addition of GA₃ or BA to the medium induced flowering of albino plants exposed to red light. The functional interaction of photoreceptors in the flowering control is discussed.     

Photoperiod affects the growth and development of yam plantlets obtained by in vitro propagation

The effects of photoperiod on the development of in vitro grown plantlets of yam (Dioscorea alata L.), were investigated. Plantlets were transplanted into pots, acclimatizated until they reached vegetative stages V₁ (3 leaves) or V₂ (8 leaves), and then grown under 12-h or 16-h photoperiod. The formation and development of underground tubers was only induced under 12-h photoperiod. Tuber initiation was not related to the initial vegetative stage of plants, and the tubers were visible at about 18 – 24 d. On the contrary, a 16-h photoperiod inhibited tuber formation and stimulated vine and leaf growth. The total dry matter production and the number of leaves per plant of V₁ stage plants were 50 and 30 % lower respectively, after 44 d under 12-h compared to 16-h photoperiod. These parameters were not influenced by photoperiod in V₂ stage plants. Consequently, the effect of 12-h photoperiod on dry matter of V₁ plants was attributed to a source limitation related to the early initiation of tuberization. The transfer of plants grown under 12-h to 16-h photoperiod stopped tuber growth and starch accumulation. On the other hand, it stimulated the shoots and the roots to grow.Abbreviations

Cytokinin Profiles of AtCKX2-Overexpressing Potato Plants and the Impact of Altered Cytokinin Homeostasis on Tuberization In Vitro

Genes encoding cytokinin oxidase/dehydrogenase (CKX) enzymes have been used lately to study cytokinin homeostasis in a variety of plant species. In this study AtCKX2-overexpressing potato plants were engineered and grown in vitro as a model system to investigate the effects of altered cytokinin levels on tuber formation and tuber size. Protein extracts from shoots and roots of transformed potato plants exhibited higher CKX activity compared to control plants. Total endogenous cytokinin levels were generally not decreased in AtCKX2 overexpressors. However, levels of bioactive cytokinins were markedly lowered, which was accompanied by increased levels of O- and N-glucosides in some transgenic lines. The AtCKX2-overexpressing plants displayed reduced shoot growth but other symptoms of the ??cytokinin deficiency syndrome?? were not recorded. The transgenic plants were able to produce tubers in noninducing conditions. In inducing conditions they developed larger tubers than control. Tubers were also formed on a greater portion of the analyzed AtCKX2 plants, but with a lower number of tubers per plant compared to control. Taken together, our data suggest that cytokinins cannot be regarded simply as positive or negative regulators of tuberization, at least in vitro. Interactions with other plant hormones that play an important role in control of tuberization, such as gibberellins, should be further studied in detail.