Preventing chimerism in potato (Solanum tuberosum L.)

Summary A fast neutron irradiation experiment using two different dose rates has been performed on the potato variety Desiree in order to obtain chimera-free mutants. Both tubers and tuber parts (cross sections) were de-eyed at different moments before and after irradiation. Adventitious sprouts successively developing on the callus in the holes of excised eyes were separated from the treated tubers and planted individually. So called grouped sprouts were separated. Results indicated that (practically) chimera-free mutants can be obtained by de-eying tuber parts shortly before irradiation, by applying relatively high dose-rates and by taking adventitious sprouts which develop not earlier than three months after irradiation.Separation of the grouped sprouts demonstrated that each part originated from a single original cell.This article is dedicated to Dr F. P. Ferwerda who till September 1969 was in charge of potato mutation research at our Institute.     

Evaluation of seedling selection for salinity tolerance in potato (Solanum tuberosum L.)

Summary Salt-tolerance in potato (Solanum tuberosum L.) was selected on the basis of germination and survival of seed sown in trays of perlite suspended in either 75 or 150 mM NaCl. Salinity reduced the germination of seed. Genetic differences in salt-tolerance were apparent with salt reducing germination more in seed collected from cv. Cara than in that collected from cv. Maris Piper. Progeny from the seedling selection were then grown to maturity to produce tubers.The relative tolerance of the parental cultivars and of unselected and selected progeny to long-term exposure to salinity was examined in a pot experiment in which plants were irrigated with either fresh water or 50 mM NaCl solution from one week after plant emergence. In this experiment, salinity significantly reduced leaf conductance, total dry matter production and partitioning of assimilate to tubers. Salinity reduced dry matter production and assimilate partitioning to tubers to a greater extent in Cara than in M. Piper. Progeny selected for short-term salt-tolerance did not exhibit greater long-term salt-tolerance than unselected progeny, and both were more sensitive than M. Piper.These results demonstrate genetic variation in salt-tolerance in potato. However, although there was a correlation between the performance of the parent to long-term salinity and survival of progeny in the seedling selection, there was no correlation between short- and long-term salt-tolerance. This suggests that characters underlying short-term tolerance may contribute to long-term tolerance but do not of themselves confer long-term tolerance. Future progress in selecting for improved salt-tolerance depends on understanding the effects of salinity on the physiological processes underlying growth and carbon partitioning.     

Divergent strategies displayed by potato (Solanum tuberosum L.) cultivars to cope with soil drought

The physiological responses of potato (Solanum tuberosum L) cultivars to soil drought at the tuberization phase and their impact on agronomically important traits were investigated in potted plants under semi‐controlled conditions. Genotype‐dependent responses of potato to water deficiency were evaluated on two pairs (tolerant/sensitive) of Polish cultivars. Each pair of cultivars had a similar genetic background but was extremely different in terms of drought tolerance evaluated on the basis of loss of tuber yield under field conditions. The results clearly indicate different mechanisms of tolerance to water deficiency and the ability to recover from soil drought in two tolerant but genetically unrelated cultivars. When subjected to soil drought, the cultivar Gwiazda had low rates of transpiration and photosynthesis and low levels of stomatal conductance due to hypersensitivity to ABA, but its maximal photochemistry efficiency and PSII performance index were unchanged. Another strategy was displayed by the dehydration‐avoidant cultivar Tajfun, which kept the stomata partially open. Thus, the plants were able to retain a relatively high rate of photosynthesis over transpiration. The parameters measured for cultivar Tajfun for photosynthesis and transpiration were the same after plant rewatering, similar to the control plants. This was not the case for the cultivar Gwiazda. The ability of plants to regenerate after soil drought relief appears to be a good indicator of potato susceptibility to soil drought and allows the yield of potato tubers to be predicted. The physiological traits identified in closely related potato cultivars but differed in their drought tolerance seem to be useful for genetic engineering and breeding programmes.     

Association mapping of quality traits in potato (Solanum tuberosum L.)

In this paper, we describe the assessment of linkage disequilibrium and its decay in a collection of potato cultivars. In addition, we report on a simple regression based association mapping approach and its results to quality traits in potato. We selected 221 tetraploid potato cultivars and progenitor lines, representing the global diversity in potato, with emphasis on genetic variation for agro-morphological and quality traits. Phenotypic data for these agro-morphological and quality traits were obtained from recent trials performed by five breeding companies. The collection was genotyped with 250 AFLP^® markers from five primer combinations. The genetic position of a subset of the markers could be inferred from an ultra dense potato map. Decay of linkage disequilibrium was estimated by calculating the squared correlation between pairs of markers using marker band intensities. Marker-trait associations were investigated by fitting single marker regression models for phenotypic traits on marker band intensities with and without correction for population structure. The paper illustrates the potential of association mapping in tetraploid potato, because existing phenotypic data, a modest number of AFLP markers, and a relatively simple statistical analysis, allowed identifying interesting associations.     

An assessment of morphogenic and transformation efficiency in a range of varieties of potato (Solanum tuberosum L.)

Summary To provide a truly genotype-independent transformation system, it is necessary to be able to transform a wide range of potato genotypes. The ability to regenerate shoots in vitro was determined for 34 potato varieties using tuber disc explants. Following a culture regime used extensively in previous studies with the variety Desiree, half of the varieties could be regenerated from tuber discs and half could not. From a sample of varieties that could be regenerated from tuber discs, all but one variety gave transgenic plants. Twelve varieties were evaluated for the capacity to regenerate shoots from leaf and internode explants excised from in vitro grown plants. All of the varieties tested regenerated adventitious shoots. Leaf and internode explants from 5 varieties were subsequently used for transformation, and transgenic plants were produced from two potato varieties that did not give transgenic plants from tuber disc explants. Some varieties could not be transformed by either method, and will require modification of the in vitro regeneration and transformation system to be successful.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - GA₃ gibberellic acid - IAA indole-3-acetic acid     

Genetic analyses for bioactive potential in tetraploid potato (Solanum tuberosum L.)

A total of four traits related to bioactive potential of the tetraploid potato were studied in a North Carolina II cross‐experiment. The objectives of this study were to estimate genetic parameters for target bioactive compounds in a tetraploid potato population, to determine phenotypic and genotypic correlations and to calculate combining ability effects of cultivars and breeding lines included in the crossing scheme. Total monomeric anthocyanins, phenolics, carotenoids and hydrophilic antioxidant capacity were measured by spectrophotometric methods. The sources of variation female (GCA_f), male (GCA_m) and their interaction (SCA) were significant for bioactive compounds and antioxidant capacity. Narrow‐sense heritability estimates (h²) were moderate for all the traits (0.413–0.657). Both phenotypic and genotypic correlations were highly significant for phenolic compounds and antioxidant capacity. As regards combining ability, positive and negative GCA and also SCA effects have been identified for bioactive compounds and antioxidant capacity. These results indicate that hybrid breeding lines can be obtained by choosing parents with significant GCA, whose bioactive potential can also be further enhanced if the crosses have a significant SCA.     

Integration of transgenes into sexual polyploidization schemes for potato (Solanum tuberosum L.)

Most potato transgenic research has focused on development of resistance to pathogens and modification of potato physiology. Many transgenes, particularly those conferring pathogen resistance, could substantially lower potato production costs in developing countries. However, transgenes have not been reported in sexually propagated 4x-2x potato hybrids commonly grown in developing countries. Two transgenes,the Bacillus thuringiensis cry3Aa endotoxin protein gene and the PVY°coat protein gene, were engineered intodiploid and tetraploid potato using Agrobacterium tumefaciens-mediated transformation. Cry3Aa was produced at high levels in several lines while the PVY° coat protein was not expressed. Diplandroid and tetraploid genotypes were crossed to produce transgenic 4x-2x hybrids. Genetic transformation had no discernable effect on fertility ofthe primary transformants, germination of4x-2x seed derived from the transformants and agronomic performance(tuber set, average tuber weight and total tuber yield) of the 4x-2xhybrids. The transgenic 4x-2xhybrids produced non-viable pollen and could only be crossed as female parents. Results suggest that transgenes, such ascry3Aa, could be expressed in 4x-2x hybrids to lower costs of production with no significant effect on plant phenotype. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic characterisation of tetraploid potato (Solanum tuberosum L.) emphasising genetic control of total glycoalkaloid content in the tubers

Two segregating resource populations were constructed, one of 216 clones using the tetraploid clones Cara and LT7 as parents and one of 176 clones using the tetraploid clones NT8 and LT7 as parents. The populations were used, in replicates, to estimate genetic parameters for total glycoalkaloid (TGA) content in the tubers, and for other agronomic and morphological traits such as tuber weight, dormancy and plant height. In both populations, the parental clones differed markedly in TGA-content and the progeny population was normally distributed for this trait after logarithmic transformation. Broad sense heritability estimate of TGA-content was 0.54 in Cara × LT7 and 0.50 in NT8 × LT7 and the trait proved to be inherited in a non-dominant manner. The minimum number of genes contributing to TGA-content was estimated to be between 3 and 7. None of the other traits recorded in this study showed a statistically significant genetic association with TGA-content. This suggests that tuber TGA-content may be genetically modulated without any significant adverse effects on other agronomic traits. The genetic parameters estimated in this study indicate that these populations are suitable for DNA-marker analysis for TGA-content in tubers. This revised version was published online in August 2006 with corrections to the Cover Date.     

Genomic prediction for yields,processing and nutritional quality traits in cultivated potato (Solanum tuberosum L.)

Current potato breeding approaches are hampered by several factors including costly seed tubers, tetrasomic inheritance and inbreeding depression. Genomic selection (GS) demonstrated interesting results regardless of the ploidy level, and can be harnessed to circumvent these problems. In this work, three GS models were evaluated using 50,107 informative SilicoDArT markers and 11 traits in two values for cultivation and use (VCU) potato trials. Two key breeding problems modelled included predicting the performance of (i) new and unphenotyped clones (cross‐validation) and (ii) a VCU using another as training set (TS). GS models performed comparably. Cross‐validation accuracy was high for D35, D45, DMW and BVAL, in ascending order. Prediction accuracies of the VCUs were highly correlated, but the best prediction was obtained for the smaller VCU using the bigger as TS. Cross‐validation and VCU prediction accuracies were higher when bigger TSs were used. The findings herein indicate that GS can be attractively integrated in potato breeding, particularly in early clonal generations to predict and select for traits with low heritability which would otherwise require more testing years, environments and resources.     

Induction of fertile flowers in potato (Solanum tuberosum L.) by silver thiosulphate anionic complex

Summary The effect of gibberellic acid containing mixtures, silver thiosulphate and extended photoperiod on flowering induction in 16 non-flowering potato genotypes and on flowering enhancement in 14 normally potato flowering genotypes was studied in sub-tropical plains of India during short-day autumn crop season of 2000–2001 and 2001–2002. Extended photoperiod alone was not successful in induction of flowering. Silver thiosulphate in combination with extended photoperiod effectively induced flowering in 16 potato genotypes studied for flowering induction. Induced flowers of some genotypes were male fertile. Normal berry setting was observed on induced flowers and seeds obtained from such berries germinated normally. Gibberellic acid containing treatments were not very effective in flower induction as they induced some flowering only in few genotypes. In the normally flowering genotypes silver thiosulphate enhanced maximum flowering and duration of flowering to a great extent.     

Breeding for improved drought tolerance in Chickpea (Cicer arietinum L.)

Chickpea (Cicer arietinum L.) is an important legume crop as a protein source across the world. It is mostly grown on arid and marginal lands where it faces drought stress at different growth stages. Drought stress exerts drastic effects on nutrient uptake, hinders the nodule formation and adversely affects yield and yield components. Generally drought at any growth stage and organizational level is responsible for reduction in economic yield. Significant variability in chickpea germplasm is present on the basis of responses to drought stress in the form of drought escape, drought avoidance and drought tolerance; these mechanisms prevent chickpea crop from harmful effects of drought. Improvement in chickpea germplasm against drought stress could be made by using several breeding approaches, that is introduction, hybridization, mutation breeding, marker‐assisted breeding and omic techniques. These breeding approaches, especially marker‐assisted breeding and omics, are further strengthened with the availability of the chickpea genome sequence. This review highlighted the significance, status and advances in different breeding strategies for improvement of drought tolerance in chickpea.     

Analysis of an incomplete diallel cross among three ssp. tuberosum varieties and seven long-day adapted ssp. andigena clones of the potato (Solanum tuberosum L.)

Summary The main aim of the experiment was to study the behaviour of seven long-day adapted clones of Solanum tuberosum ssp. andigena (A) and three varieties of S. tuberosum ssp. tuberosum (T) in an incomplete diallel cross. The four groups of crosses, TxT, TxA, AxT and AxA, held 2, 12, 20 and 21 populations, respectively. Eleven of the intersubspecific crosses were exact reciprocals. Thirty random clones per population were grown as second clonal generation in an incomplete block design with four replications in plots of two plants per clone in 1980, the parents included in many plots. Data were recorded on ten characters.The results showed that group TxA gave the highest tuber yield and AxT the second highest. The F-values indicated highly significant genotypic effects for all characters in all populations. The mean genotypic coefficients of variation (GCV), were the highest for group AxA for all characters. The ANOVA, based on population means showed significant mean squares for general combining abilities (GCA) and for differences of reciprocalls for all characters; the specific combining ability (SCA) mean squares were significant only for date of emergence, plant height and haulm type. Almost all GCA effects of the parents were significant positive or negative. Several populations showed also significant SCA effects. The correlations between the GCA effects and the parental values were of about the same magnitude as the heritabilities; those between the population means and the midparental values were slightly lower.Compared to the midparental values, most populations showed significant positive heterosis for date of emergence, haulm type, number of tubers and under water weight; all populations had significant negative heterosis for mean tuber weight and nearly all for plant height and general impression. All populations from groups TxT and AxA had significant negative heterosis for tuber yield and so had 13 out of the 20 populations from group AxT, while seven of the 12 from group TxA showed significant positive and four significant negative heterosis. The average heterosis for tuber yield of group TxT was –11%, of TxA 3.4%, of AxT–9% and of AxA–18%. The highest mean tuber yields were obtained from crosses with a very high yielding A-parent as female, while the heterosis was negative. Though significant differences between the 11 reciprocals occurred for all characters, consistent differences occurred only for tuber yield. Averaged over the 11 crosses the TxA populations outyielded their exact reciprocals by 10.7%. For the highest tuber yields in A-T crosses, therefore, the tuberosum cytoplasm seems to be preferable. By contrast for a high male fertility the andigena cytoplasm is preferable.     

Identification of epistatic interaction affecting glycoalkaloid content in tubers of tetraploid potato (Solanum tuberosum L.)

Glycoalkaloids are secondary metabolites characterised by an undesirable taste, and known to be toxic when consumed in large quantities. Their content in potato tubers should therefore be selected against and DNA markers can significantly facilitate such a process. This study was designed to describe the genetic control of the total content of glycoalkaloids (TGA) in the potato tuber and to enhance the development of DNA markers. A segregating population was constructed from tetraploid potatoclones, which were characterised by divergent TGA-content. The population had a high-TGA parent, LT7, and a low-TGA parent, NT8. We used 342 Inter-Simple Sequence Repeat (ISSR) single primers or two such primer combinations and 36 Cleaved Amplified Polymorphic Sequence (CAPS)primer pairs, digested with various restriction enzymes, to amplify random PCR products. Parental clones of both populations were used to identify polymorphic PCR products for further linkage analyses with TGA-content. In these analyses, one single ISSR marker was found to be significantly associated with TGA-content. A multiple regression analysis was also carried out using a ‘stepwise’ procedure. In this regression analysis TGA-content was the dependent variable whereas the polymorphic PCR products and all possible two-way interactions among them were the independent variables. The resulting best model consisted of an interaction between two loci in addition to a single locus effect. This interaction suggests that the expression of TGA-content is partially modulated by two interacting loci. This revised version was published online in July 2006 with corrections to the Cover Date.     

Assessment of genetic and phenotypic variation among intraspecific somatic hybrids of potato, Solanum tuberosum L.

Intraspecific somatic hybrids have been produced by protoplast fusion in eight combinations involving 10 dihaploids (2n= 2x= 24) in an attempt to provide new material for potato breeding. Cytological analysis revealed extensive variation in chromosome number, such as aneuploid, aneusomatic and mixoploid hybrids. Most of the hybrids represented the expected chromosome number of 48; however, the frequency of aneuploids reached 50% in some combinations. Some hybrids carried structurally rearranged chromosomes and exhibited a high frequency of aberrant anaphases. Isozyme and random amplified polymorphic DNA (RAPD) patterns of the hybrids from the same fusion combination were uniform. In the field, somatic hybrids showed wide phenotypic variation in 20 morphological characters. There was a significant correlation between certain leaf characters and the ploidy level, which may be used to distinguish the tetraploid hybrids from hexaploids and octoploids. Tuber yield and flowering intensity were highest in tetraploid hybrids (2n= 4x= 48). Eighteen of the 73 hybrids reached higher yields than the standard variety ‘Adretta’. Floral development and fertility were restored in hybrids derived from fusions between non-flowering or sterile dihaploids.     

Impact of nitrogen supply on leaf water relations and physiological traits in a set of potato (Solanum tuberosum L.) cultivars under drought stress

Improved adaptation of potato to limited water availability is needed for stable yields under drought. The maintenance of the cell water status and protection of cellular components against dehydration are important for drought tolerance, and the N status of plants affects the regulation of various respective metabolic processes. A 2‐year pot trial with 17 potato cultivars was conducted under a rain‐out shelter including two water regimes and two N‐levels to investigate genotypic differences concerning osmotic adjustment (OA) and relevant biochemical traits in relation to nitrogen (N) supply. Drought stress resulted in a rapid decrease in the leaf osmotic potential. The N, protein and proline contents increased under drought, while the N protein/N_Kjeldahl ratio decreased. Initially, total soluble sugars increased at both N‐levels but dropped back to the control level at high N‐availability under prolonged drought while remaining high in N‐deficient plants. Results indicate that potatoes have only a limited capacity of active OA and that increasing sugar and proline concentrations are rather associated with the protection of cellular components. High N supply promoted the N protein/N_Kjeldahl ratio at short‐term drought and enhanced proline accumulation. Significant genotypic differences were observed for all investigated traits.     

Somaclonal variation for resistance to Verticillium dahliae in potato (Solanum tuberosum L.) plants regenerated from callus

Summary Plant tissue culture is recognized as an important tool to generate useful genetic variability for crop improvement. Regenerated plants from callus induced from stem explants of Solanum tuberosum cv Désirée were assessed by in vitro selection, for resistance to Verticillium dahliae. This fungus is the causal agent of Verticillium wilt, a serious vascular wilt disease both in crops and wild species.The rate of in vitro multiplication by single node cuttings was used as a parameter of screening in two selection cycles with different concentrations of V. dahliae filtrate. One resistant clone was selected and then evaluated by inoculation in the growth chamber. Induced damage, and morphological traits (dry weight, leaf area and tuber production) were estimated. The selected clone was comparable to the resistant control, cv Kondor.The results suggest that genetic variation induced in tissue culture cound be utilized to generate disease resistance.     

Optimising potato breeding. I. The genotypic,environmental and genotype-environment coefficients of variation for tuber yield and other traits in potato (Solanum tuberosum L.) under different experimental conditions

Summary Coefficients of variation for genotypic differences, genotype-location, genotype-year and genotype-location-year interaction were computed from reported data of potato trials in order to use them in a decision support system. The obtained coefficients of variation not only varied considerably between traits, but also between trials within traits. It is argued that only parameter values representative for the pertaining breeding situation should be used for optimising a selection programme.     

A comparative assessment of DNA fingerprinting techniques (RAPD, ISSR, AFLP and SSR) in tetraploid potato (Solanum tuberosum L.) germplasm

Several DNA marker systems and associated techniques are available today for fingerprinting plant germplasm but information on their relative usefulness in particular crops is limited. The study investigated PCR based DNA fingerprinting in a set of 39 potato cultivars using RAPDs (20 primers), ISSRs (6 primers), AFLPs (2 primers) and SSRs (5 primer pairs). Results show that each of the four techniques can on their own, individually identify each cultivar, but that techniques differ in the mean number of profiles generated per primer (or primer pair) per cultivar, referred to as Genotype Index (GI). The order of merit based on this criterium and in this material was AFLPs (GI = 1.0), a multi-locus SSR (GI = 0.77),RAPDs (GI = 0.53), ISSRs (GI = 0.47) and single locus SSRs (GI = 0.36). Problems in relating banding patterns to individual loci and alleles for polyploid genomes, using these techniques as they are currently employed, are also discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Molecular breeding for resistance to Phytophthora infestans (Mont.) de Bary in potato (Solanum tuberosum L.): a perspective of cisgenesis

Late blight caused by Phytophthora infestans is one of the most devastating diseases in potato cultivation and is mostly controlled by the application of chemicals. However, introduction of combinations of resistance ( R ) genes conferring broad-spectrum resistance from wild Solanum species into cultivated potatoes is considered the most practical and promising approach to achieve durable resistance. This can be realized via classical breeding or genetic modification (GM). Because classical breeding is very time-consuming and is often hampered by linkage drag, a GM approach seems logic in this heterozygous and vegetatively propagated crop. During the last decades, many R genes have been identified in several wild Solanum species. Some have been cloned and more will follow. When these genes are derived from species crossable with cultivated potato (so-called cisgenes), application in resistance breeding using a GM approach is similar to an introgression breeding approach, in that the exploited genes are indigenous to the crop. Pending deregulation or derogation of cisgenesis, the use of cisgenic R genes would be an ideal strategy to accomplish durable resistance in potato.