Transgenic Potato Research Articles

An Assay for Express Screening of Potato Transformants by GFP Fluorescence

An express assay for screening of potato transformants by their GFP fluorescence intensities is developed. In comparison to the widely used methods of transgenic plant screening by PCR, Real-Time RTPCR or Northern-blotting, the GFP fluorescence assay needs no expensive reagents and takes less time. This approach may also be used for nondestructive screening of the T0 transgenic regenerants which can be further grown and used. To prove this assay reliability, the expression of the hGFP gene in the leaves of transgenic potato (cv. Skoroplodny) plants, determined by its mRNA accumulation, was compared to GFP fluorescence intensity in the micro-samples of aseptic plant leaves. The strong correlation between the results of these two methods is the evidence of positive dependence of GFP fluorescence intensity on the target mRNA content.

Lateral Root Development in Potato Is Mediated by Stu-mi164 Regulation of NAC Transcription Factor.

The NAC designation is derived from petunia (Petunia hybrida) gene NO APICAL MERISTEM (NAM) and Arabidopsis genes ATAF1/ATAF2 and CUP-SHAPED COTYLEDON2 (CUC2), which belongs to the family of plant-specific transcription factors (TFs), and plays important role in plant development processes, such as response to biotic and abiotic stress, and hormone signaling. MicroRNAs (miRNAs) are a class of small, non-coding endogenous RNAs which play versatile and significant role in plant stress response and development via negatively affecting gene expression at a post-transcriptional level. Here, we showed that Stu-mi164 had a complementary sequence in the CDS sequence of potato NAC TFs, and that NAC expression exhibited significant differences under osmotic stress. We measured expression levels of the Stu-mi164 target gene StNAC262 between control and PEG-treated plants using real-time PCR, and the results demonstrated that they had inverse relationship. We suggested that Stu-miR164 might drive overexpression of NAC gene under osmotic stress in potato. To confirm the regulation of NAC TFs by Stu-mi164, we developed transgenic plants, using Agrobacterium tumefaciens–mediated transformation, of the potato cultivars “Gannongshu 2” and “Kexin 3” overexpressing the Stu-mi164 or the TF StNAC262. Real-time PCR analysis of transgenic potato plants under osmotic (PEG) stress, showed that potato plants overexpressing Stu-mi164 had reduced expression of StNAC262 and their osmotic resistance decreased. Furthermore, these plants had low number of lateral roots although the same length as the control. Our findings support the regulatory role of Stu-miRNAs in controlling plant response to osmotic stress via StNAC262.

Highly Efficient Agrobacterium-Mediated Transformation of Potato (Solanum tuberosum) and Production of Transgenic Microtubers.

The following method enables the rapid production of transgenic potato plants and microtubers for gene validation and expression, or promoter studies. The method is highly efficient, with reproducible transformation efficiencies of at least 50% to 60% with potato cv. Desiree, and can produce transgenic microtubers within 6 months of initiation of the experiment. Microtubers are produced in the absence of hormones, giving an in vitro gene testing system broadly analogous to the natural state. © 2018 by John Wiley & Sons, Inc.

Development of Real-time PCR Assays for the Detection of the pin II Terminator (tpinII) Used in GM Constructs and Its Donor Organism, Potato (Solanum tuberosum)

Two singleplex TaqMan methods were developed for the detection of potato targets: one for the detection of the tpinII terminator, which is an emerging terminator used in GM constructs, and one for the detection of the endogenous StLS gene of potato. Performance criteria such as specificity and sensitivity were successfully tested for the two methods, taking into account the recommendations of international guidelines. The presence of the StLS target was checked in 16 potato cultivars. The StLS target is present at low copy number and can be used for quantitation purposes, as demonstrated on transgenic potatoes in this paper. The StLS target is an excellent candidate to replace the presently recommended endogenous target based on the UGP gene, which shows several disadvantages due to its high copy number and lack of specificity. The research also indicates that DNA can easily be extracted from different parts of potato tubers with a classical cetyltrimethylammonium bromide method.

Enhanced Fusarium oxysporum f. sp. tuberosi Resistance in Transgenic Potato Expressing a Rice GLP Superoxide Dismutase Gene

Germin like proteins (GLPs) are a large group of related and ubiquitous plant proteins which are considered to be involved in different processes important for plant development and defense. Multiple functional copies of this gene family have been reported in a number of species (wheat, barley, rice, soybean mosses and liverwort), and their role is being evaluated by gene regulation studies and transgenic approaches. To analyze the role of a rice (Oryza sativa) root expressed germin like protein1 OsRGLP1, for its antifungal activity, transgenic potato plants were developed. These transgenic potato plants were molecularly characterized and biologically assessed after inoculation with Fusarium oxysporum f. sp. tuberosi. Functional analysis showed high accumulation of H2O2, increased Superoxide Dismutase (SOD) activity and no oxalate oxidase activity (OxO) in transgenics in comparison to nontransformed control. This increased SOD activity, resistance to heat and sensitivity to H2O2 suggest it is a Fe-like SOD. OsRGLP1 expression in potato plants exhibited enhanced resistance in comparison to nontransformed wild type plants suggesting its role in providing protection against Fusarium oxysporum f. sp. tuberosi through elevated SOD level. Overall, results suggest that OsRGLP1 is a candidate for the engineering of potato plants with increased fungal tolerance however, the greater height and tuber number was observed. This phenotype associated with the resistance needs to be evaluated to determine if this is a positive or negative feature.

Orange: a target gene for regulating carotenoid homeostasis and increasing plant tolerance to environmental stress in marginal lands.

Carotenoids play essential roles in various light-harvesting processes in plants and help protect the photosynthetic machinery from photo-oxidative damage. Orange genes, which play a role in carotenoid accumulation, have recently been isolated from several plant species, and their functions have been intensively investigated. The Orange gene (IbOr) of sweet potato [Ipomoea batatas (L.) Lam] helps maintain carotenoid homeostasis to improve plant tolerance to environmental stress. IbOr, a protein with strong holdase chaperone activity, directly interacts with phytoene synthase, a key enzyme involved in carotenoid biosynthesis, in plants under stress conditions, resulting in increased carotenoid accumulation and abiotic stress tolerance. In addition, IbOr interacts with the oxygen-evolving enhancer protein 2-1, a member of a protein complex in photosystem II that is denatured under heat stress. Transgenic sweet potato plants overexpressing IbOr showed enhanced tolerance to high temperatures (47 °C). These findings indicate that IbOr protects plants from environmental stress not only by controlling carotenoid biosynthesis, but also by directly stabilizing photosystem II. In this review, we discuss the functions of IbOr and Or proteins in other plant species and their possible biotechnological applications for molecular breeding for sustainable development on marginal lands.

PAMP-responsive ATL gene StRFP1 and its orthologue NbATL60 positively regulate Phytophthora infestans resistance in potato and Nicotiana benthamiana

Ubiquitination is a post-translational modification that plays a crucial role during the regulation of plant immune signalling. The plant ATL family consists of a large number of putative RING type ubiquitin ligases. We show that potato ATL family gene StRFP1 and its orthologue NbATL60 from N. benthamiana both respond to Phytophthora infestans culture filtrate (CF) and flg22 induction. StRFP1 positively regulates immunity against P. infestans in potato. Ectopic transient expression of StRFP1 or expression of NbATL60 in N. benthamiana also enhances late blight resistance. By contrast, silencing NbATL60 in N. benthamiana reduces late blight resistance and leads to plant growth inhibition. Both StRFP1 and NbATL60 localize to the plasma membrane and intracellular puncta and possess E3 Ligase activity in vitro. Furthermore we demonstrate that the RING finger domain mutants of StRFP1 and NbATL60 lost E3 ligase activity and fail to suppress P. infestans colonization in N. benthamiana, indicating that E3 ligase activity is critical for StRFP1 and NbATL60 to regulate immunity. Overexpression or RNA interference of StRFP1 in transgenic potato led to increased or decreased expression of PTI maker genes (WRKY7, WRKY8, ACRE31 and Pti5) respectively. Similarly silencing of NbATL60 in N. benthamiana decreases expression of these PTI marker genes. Moreover, VIGS of NbATL60 in N. benthamiana did not compromise P. infestans PAMP INF1 or R2/Avr2, R3a/AVR3a, Rx/Cp and Pto/AvrPto triggered cell death. These results indicate that ATL genes StRFP1 and NbATL60 contribute to basal immunity (PTI) in Solanaceous plants.

Functional analysis of StDWF4 gene in response to salt stress in potato

The DWARF4 (DWF4) gene encodes a C-22 hydroxylase which is pivotal for brassinosteroids (BRs) biosynthesis. In this research, aimed to understand the molecular mechanism of DWF4 on regulation of potatoes tolerance to salt stress, DWF4 was cloned from potato, named as StDWF4. Its 1476 bp open reading frame encodes a protein of 491 amino acids. The StDWF4-overexpressing (OE) and interference-expressing (RNAi) transgenic potato plants were acquired using Agrobacterium-mediated transformation, respectively. Tissue specific analysis using Quantitative real-time polymerase chain reaction (qRT-PCR) demonstrated that the StDWF4 gene expressed in the leaves, stems and roots of the transgenic and un-transgenic (NT) plants, with specially increased (StDWF4-OE)/reduced (StDWF4-RNAi) expression in the roots. The content of malondialdehyde (MDA) in StDWF4-OE potato plants was lower than that of NT, and proline content was higher than that of NT. MDA and proline content in StDWF4-OE and NT under salt-stress was significantly higher than that of the control and was increased at different sampling times. The content of soluble protein, soluble sugar and the activities of superoxide dismutase (SOD), peroxidase (POD) and ascorbate peroxidase (APX) was higher in the StDWF4-OE plantlets at varied salt treatment time than in the NT potatoes. Reduction of H2O2 content in the StDWF4-OE plants was observed. All above plant physiology indicators in the StDWF4-RNAi potatoes showed opposite variation trends. The results proved that the overexpressing of StDWF4 in potato plantlets can enhance the salt resistance by alleviating the negative effects of salt-stress. However, its interference expression in potato plants depresses the salt resistance. The results lay the groundwork for intensive study of BRs regulation in potato growth and development, and will help us to reveal the molecular mechanisms of how the BRs signaling regulate potato salt tolerance.

Nicotiana benthamiana Matrix Metalloprotease 1 (NMMP1) gene confers disease resistance to Phytophthora infestans in tobacco and potato plants

We previously isolated Nicotiana benthamiana matrix metalloprotease 1 (NMMP1) from tobacco leaves. The NMMP1 gene encodes a highly conserved, Zn-containing catalytic protease domain that functions as a factor in the plant’s defense against bacterial pathogens. Expression of NMMP1 was strongly induced during interactions between tobacco and one of its pathogens, Phytophthora infestans. To elucidate the role of the NMMP1 in defense of N. benthamiana against fungal pathogens, we performed gain-of-function and loss-of-function studies. NMMP1-overexpressing plants had stronger resistance responses against P. infestans infections than control plants, while silencing of NMMP1 resulted in greater susceptibility of the plants to the pathogen. This greater susceptibility correlated with fewer NMMP1 transcripts than the non-silenced control. We also examined cell death as a measure of disease. The amount of cell death induced by the necrosis-inducing P. infestans protein 1, PiNPP1, was dependent on NMMP1 in N. benthamiana. Potato plants overexpressing NMMP1 also had enhanced disease resistance against P. infestans. RT-PCR analysis of these transgenic potato plants revealed constitutive up-regulation of the potato defense gene NbPR5. NMMP1-overexpressing potato plants were taller and produced heavier tubers than control plants. We suggest a role for NMMP1in pathogen defense and development.

Immune analysis of cry1Ab-genetically modified potato by in-silico analysis and animal model.

The safety of feeding transgenic potato (Solanum tuberosum), resistant to the potato tuber moth, to Wistar rats was examined from an immunological perspective. The genetically modified potato (GMP) was harbouring cry1Ab and nptII genes as target and selectable marker genes, respectively. In-silico analysis reconfirmed that Cry1Ab and NPTII protein sequences have no significant homology to known toxins or known allergens. The Wistar rats were fed a diet containing 20% GMP or its parental control, non-GMP (NGMP), for 90days. The consumption of GMP food did not affect the growth rate, food intake, food efficiency, and general health status of the rats. There were no significant differences in the serum concentrations of immunoglobulin A (IgA), IgG, IgM, interleukin 6 (IL-6), and IFN-γ between GMP and NGMP-fed rats. Based on such data, it is concluded that the transgenic potato had no adverse effect on immunity functions of Wistar rats.

The plasma membrane H+-ATPase gene family in Solanum tuberosum L. Role of PHA1 in tuberization.

This study presents the characterization of the plasma membrane (PM) H+-ATPases in potato, focusing on their role in stolon and tuber development. Seven PM H+-ATPase genes were identified in the Solanum tuberosum genome, designated PHA1-PHA7. PHA genes show distinct expression patterns in different plant tissues and under different stress treatments. Application of PM H+-ATPase inhibitors arrests stolon growth, promotes tuber induction, and reduces tuber size, indicating that PM H+-ATPases are involved in tuberization, acting at different stages of the process. Transgenic potato plants overexpressing PHA1 were generated (PHA1-OE). At early developmental stages, PHA1-OE stolons elongate faster and show longer epidermal cells than wild-type stolons; this accelerated growth is accompanied by higher cell wall invertase activity, lower starch content, and higher expression of the sucrose-H+ symporter gene StSUT1. PHA1-OE stolons display an increased branching phenotype and develop larger tubers. PHA1-OE plants are taller and also present a highly branched phenotype. These results reveal a prominent role for PHA1 in plant growth and development. Regarding tuberization, PHA1 promotes stolon elongation at early stages, and tuber growth later on. PHA1 is involved in the sucrose-starch metabolism in stolons, possibly providing the driving force for sugar transporters to maintain the apoplastic sucrose transport during elongation.

Expression of potato virus S gRNA fragments in transgenic potato plants triggers RNA-interference against related and unrelated viruses

This chapter reviews the discoveries and initial characterizations (1930–1990) of three plant rhabdoviruses, sonchus yellow net virus, potato yellow dwarf virus, and lettuce necrotic yellows virus, that have become model systems for research on this group of enveloped negative-strand RNA plant viruses. We have used our personal perspectives to review the early historical studies of these viruses, the important technologies and tools, such as density gradient centrifugation, that were developed during the research, and to highlight the eminent scientists involved in these discoveries. Early studies on sites of virus replication, virion structure, physicochemical composition, and the use of protoplasts and vector insect cell culture for virus research are discussed, and differences between the nuclear and cytoplasmic lifestyles of plant rhabdoviruses are contrasted. Finally, we briefly summarize the genome organization and more recent developments culminating in the development of a reverse genetics system for plant negative-strand RNA viruses.

Overexpression of Golgi Protein CYP21-4s Improves Crop Productivity in Potato and Rice by Increasing the Abundance of Mannosidic Glycoproteins.

CYP21-4 is a novel Golgi-localized cyclophilin protein involved in oxidative stress tolerance. Here, we generated transgenic plants overexpressing AtCYP21-4 and OsCYP21-4 in potato and rice, respectively. The stems and roots of AtCYP21-4–overexpressing potato plants were longer than those of wild-type (WT) plants, which resulted in heavier tubers. In vitro tuberization in the transgenic potato also resulted in significantly greater tuber number and weight, as well as a shorter time to microtuber formation. Similarly, OsCYP21-4–overexpressing transgenic rice plants had higher biomass and productivity with longer early-stage internodes than the WT and higher seed weight. Immunoblot analysis with CYP21-4 antibody showed that these productivity-enhancing phenotypes were associated with high CYP21-4s protein expression. Anatomically, transgenic potato stems exhibited higher lignin content in xylem cells and thicker leaves. In addition, relative content of mannosidic glycoproteins per unit of total protein was above 20% in transgenic potato tubers and rice grains. Based on these findings, we propose that CYP21-4s are involved in the growth and development of plant vegetative and storage tissues via their effects on glycoprotein abundance or glycan processing in the Golgi apparatus. Thus, increasing CYP21-4s expression in crops could represent an alternative way to increase crop productivity and yield.

R/Avr gene expression study of Rpi-vnt1.1 transgenic potato resistant to the Phytophthora infestans clonal lineage EC-1

The Avr avirulence gene of Phytophthora infestans and R gene of the potato are the genetic components of the gene-for-gene interaction resulting in host plant resistance. This effector-triggered immunity has been recently exploited to generate extreme resistance to late blight in potato by genetic engineering. The choice of the R genes, their number forming a R gene stack, and the pathogen Avr gene diversity will likely determine how long this extreme resistance will last. Here, we report on a comparative study on the Rpi-vnt1.1 gene which originated from Solanum venturii, and was introduced in the potato variety ‘Desiree’ by genetic transformation, and the Avr-vnt1 gene from two isolates of the EC-1 lineage of P. infestans. EC-1 was reported previously as not expressing the Avr-vnt1 gene and, therefore, being virulent on Rpi-vnt1.1 transgenic plants. Unexpectedly, whole-plant resistance assays identified 5 out of 52 transgenic events as resistant to two isolates of P. infestans, POX067 and POX109, belonging to the EC-1 lineage. We demonstrated that in both isolates, the Avr-vnt1.1 gene was expressed at a low level. Expression of the Rpi-vnt1.1 gene was shown to be rapidly increased by two-fold and subsequently to have steady state expression for at least 5 days after the inoculation. The Rpi-vnt1.1 gene in addition to other R genes as a stack in farmers’ preferred varieties will confer extreme resistance to late blight disease and rotations of plants with different R-gene-stack in time is likely to last longer than plants with single R gene.

Simultaneous silencing of isoamylases ISA1, ISA2 and ISA3 by multi-target RNAi in potato tubers leads to decreased starch content and an early sprouting phenotype.

Isoamylases hydrolyse (1–6)-alpha-D-glucosidic linkages in starch and are involved in both starch granule formation and starch degradation. In plants, three isoamylase isoforms with distinct functions in starch synthesis (ISA1 and ISA2) and degradation (ISA3) have been described. Here, we created transgenic potato plants with simultaneously decreased expression of all three isoamylases using a chimeric RNAi construct targeting all three isoforms. Constitutive expression of the hairpin RNA using the 35S CaMV promoter resulted in efficient silencing of all three isoforms in leaves, growing tubers, and sprouting tubers. Neither plant growth nor tuber yield was effected in isoamylase-deficient potato lines. Interestingly, starch metabolism was found to be impaired in a tissue-specific manner. While leaf starch content was unaffected, tuber starch was significantly reduced. The reduction in tuber starch content in the transgenic plants was accompanied by a decrease in starch granules size, an increased sucrose content and decreased hexose levels. Despite the effects on granule size, only little changes in chain length composition of soluble and insoluble glucose polymers were detected. The transgenic tubers displayed an early sprouting phenotype that was accompanied by an increased level of sucrose in parenchyma cells below the outgrowing bud. Since high sucrose levels promote sprouting, we propose that the increased number of small starch granules may cause an accelerated turnover of glucan chains and hence a more rapid synthesis of sucrose. This observation links alterations in starch structure/degradation with developmental processes like meristem activation and sprout outgrowth in potato tubers.

Transgenic Potato Plants Overexpressing SOD and APX Exhibit Enhanced Lignification and Starch Biosynthesis with Improved Salt Stress Tolerance

Potato (Solanum tuberosum L.) is a major crop worldwide and its productivity is severely reduced by high soil salinity. The aim of the current study was to gain insights into the response and regulation of the antioxidant system in the transgenic potato, overexpressing two genes viz. thermostable CuZn-superoxide dismutase (PaSOD) gene from polyextremophile high-altitude plant Potentilla atrosanguinea and ascorbate peroxide (RaAPX) gene from Rheum australe under salinity stress. Transgenic lines and wild type (WT) were assessed under 50, 100, and 150 mM concentrations of NaCl treatment at the anatomical, transcriptional and metabolic levels. Although superoxide dismutase (SOD) and ascorbate peroxide (APX) activities in transgenic lines were significantly higher than WT under salt stress, the secondary cell wall lignification was profoundly affected by the modulated antioxidant enzymes activities and the corresponding H2O2 levels in transgenic. Expression profiles showed that the several genes and transcription factors directly involved in lignin biosynthesis were profoundly induced in transgenic lines as compared to WT. In addition, transgenic plants harbored increased starch accumulation, improved growth attributes, and reduced accumulation of reactive oxygen species than WT plants. Together, these results indicated that the expression of SOD and APX genes in transgenic potato may function as a positive set of physiological, anatomical, and molecular adjustments in the H2O2 regulated lignin biosynthesis signaling pathway, enabling the plants to withstand severe saline conditions.

Inhibitory effect of StCYP707A1 gene on tuberization in transgenic potato

Tuber formation of potato is a complex biological process and is regulated by many factors including phytochrome, hormones, transcription factors, RNAs, microRNAs, etc. In our previous study, CYP707A1, an ABA 8′-hydroxylase gene, was down-regulated in the StCOL antisense transgenic potato stolons, but its relation with potato tuberization has not been studied yet. In this study, to investigate the role of this gene in potato tuberization, we cloned a CYP707A1 gene from potato cultivar Desiree and constructed the StCYP707A1 over-expression and anti-sense potato lines by Agrobacterium-mediated transformation. In total, four over-expression transgenic lines and nine anti-sense transgenic lines were confirmed by PCR analysis. Semi-quantitative RT-PCR and qRT-PCR analysis showed that the level of StCYP707A1 transcripts was significantly higher in the over-expression lines (A7 and A9) and lower in the anti-sense lines (F2 and F8), compared to the wild-type control. We further analyzed tuber formation in the transgenic lines and the wild-type control. The result showed that tuber yield per plant and average tuber weight were decreased in A7 and A9 and increased in F2. We also measured the content of ABA and GA3 in transgenic lines. ABA level was reduced in A7 and A9 and increased in F2. Contrariwise, the concentration of GA3 was higher in A7 and A9, and lower in F2 than in wild-type control. These results indicate that StCYP707A1 negatively affects potato tuberization through ABA regulation on gibberellic acid concentration.

Profiling of anthocyanins in transgenic purple-fleshed sweet potatoes by HPLC-MS/MS.

Anthocyanins in purple-fleshed sweet potato (PSP) are beneficial to human health. The leaf color (Lc) gene is a transcription factor involved in regulating anthocyanin biosynthesis. The anthocyanin profiles of wild-type PSP of Ayamurasaki and its three Lc-transgenic lines were investigated by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). In vitro antioxidant activities of wild-type and Lc-transgenic lines, including reducing power activity, DPPH radical scavenging activity, hydroxyl radical scavenging activity, linoleic acid autoxidation inhibition activity, ABTS free radical scavenging activity and oxygen radical absorbance capacity activity, were measured. The results showed that the total anthocyanin contents increased 1.5-1.9 times in three transgenic lines compared with that in wild-type PSP. Seventeen anthocyanins were found in wild-type PSP, while 19 in Lc-transgenic lines including cyanidin-based, peonidin-based and pelargonidin-based anthocyanins. Three pelargonidin-based anthocyanins were detected in three Lc-transgenic lines. Among them, the relative contents of cyanidin-based and pelargonidin-based anthocyanins increased 1.9-2.0 and 3.4-4.5 times respectively, while peonidin-based anthocyanins decreased 1.8-1.9 times in Lc-transgenic lines, compared with wild-type PSP. PSP from wild-type Ayamurasaki and three Lc-transgenic lines exhibited potent antioxidant activities, whereas there was no distinct difference among them. The transgene Lc significantly increased the content of total anthocyanins and remarkably changed the anthocyanin profiles in Ayamurasaki. Such novel and high content of anthocyanins obtained in the Lc-transgenic lines with potent antioxidant activities may provide unique functional products with potential helpful for human health. © 2017 Society of Chemical Industry.

A soluble starch synthase I gene, IbSSI, alters the content, composition, granule size and structure of starch in transgenic sweet potato

Soluble starch synthase I (SSI) is a key enzyme in the biosynthesis of plant amylopectin. In this study, the gene named IbSSI, was cloned from sweet potato, an important starch crop. A high expression level of IbSSI was detected in the leaves and storage roots of the sweet potato. Its overexpression significantly increased the content and granule size of starch and the proportion of amylopectin by up-regulating starch biosynthetic genes in the transgenic plants compared with wild-type plants (WT) and RNA interference plants. The frequency of chains with degree of polymerization (DP) 5–8 decreased in the amylopectin fraction of starch, whereas the proportion of chains with DP 9–25 increased in the IbSSI-overexpressing plants compared with WT plants. Further analysis demonstrated that IbSSI was responsible for the synthesis of chains with DP ranging from 9 to 17, which represents a different chain length spectrum in vivo from its counterparts in rice and wheat. These findings suggest that the IbSSI gene plays important roles in determining the content, composition, granule size and structure of starch in sweet potato. This gene may be utilized to improve the content and quality of starch in sweet potato and other plants.

Comparative statistical component analysis of transgenic, cyanophycin-producing potatoes in greenhouse and field trials.

Potatoes are a promising system for industrial production of the biopolymer cyanophycin as a second compound in addition to starch. To assess the efficiency in the field, we analysed the stability of the system, specifically its sensitivity to environmental factors. Field and greenhouse trials with transgenic potatoes (two independent events) were carried out for three years. The influence of environmental factors was measured and target compounds in the transgenic plants (cyanophycin, amino acids) were analysed for differences to control plants. Furthermore, non-target parameters (starch content, number, weight and size of tubers) were analysed for equivalence with control plants. The huge amount of data received was handled using modern statistical approaches to model the correlation between influencing environmental factors (year of cultivation, nitrogen fertilization, origin of plants, greenhouse or field cultivation) and key components (starch, amino acids, cyanophycin) and agronomic characteristics. General linear models were used for modelling, and standard effect sizes were applied to compare conventional and genetically modified plants. Altogether, the field trials prove that significant cyanophycin production is possible without reduction of starch content. Non-target compound composition seems to be equivalent under varying environmental conditions. Additionally, a quick test to measure cyanophycin content gives similar results compared to the extensive enzymatic test. This work facilitates the commercial cultivation of cyanophycin potatoes.