Phenotypic and transcriptomics characterization uncovers genes underlying tuber yield traits and gene expression marker development in potato under aeroponics.

MAIN CONCLUSION: Transcriptome analysis in potato varieties revealed genes associated with tuber yield-related traits and developed gene expression markers. This study aimed to identify genes involved in high tuber yield and its component traits in test potato varieties (Kufri Frysona, Kufri Khyati, and Kufri Mohan) compared to control (Kufri Sutlej). The aeroponic evaluation showed significant differences in yield-related traits in the varieties. Total RNA sequencing was performed using tuber and leaf tissues on the Illumina platform. The high-quality reads (QV > 25) mapping with the reference potato genomes revealed statistically significant (P < 0.05) differentially expressed genes (DEGs) into two categories: up-regulated (> 2 Log2 fold change) and down-regulated (< -2 Log2 fold change). DEGs were characterized by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Collectively, we identified genes participating in sugar metabolism, stress response, transcription factors, phytohormones, kinase proteins, and other genes greatly affecting tuber yield and its related traits. A few selected genes were UDP-glucose glucosyltransferase, glutathion S-transferase, GDSL esterase/lipase, transcription factors (MYB, WRKY, bHLH63, and BURP), phytohormones (auxin-induced protein X10A, and GA20 oxidase), kinase proteins (Kunitz-type tuber invertase inhibitor, BRASSINOSTEROID INSENSITIVE 1-associated receptor kinase 1) and laccase. Based on the selected 17 peptide sequences representing 13 genes, a phylogeny tree and motifs were analyzed. Real time-quantitative polymerase chain reaction (RT-qPCR) analysis was used to validate the RNA-seq results. RT-qPCR based gene expression markers were developed for the genes such as 101 kDa heat shock protein, catechol oxidase B chloroplastic, cysteine protease inhibitor 1, Kunitz-type tuber invertase inhibitor, and laccase to identify high yielding potato genotypes. Thus, our study paved the path for potential genes associated with tuber yield traits in potato under aeroponics.

Tuber quality enhancement via grafting potato onto a wooden goji rootstock through vitalizing multi-pathways.

Grafting is applied in Solanaceae to improve growth and quality traits. However, grafting potato onto a wooden goji rootstock is rare. Our study introduces a novel distant grafting technique to investigate potato scion responses, specifically regarding photosynthetic and tuber nutritional quality. The physiological and transcriptomic findings reveal an increase in photosynthesis ratio and carbon fixation in potato leaves after 45 days of grafting due to the upregulation of pivotal genes (PsbA, PPC1, rbcl, and GAPDH). After 95 days of long-term growth, the leaf redox balance was maintained with intensified chlorophyll synthesis, facilitated by the enrichment of crucial genes (GUN4, CHLH, CHLP, CAO) and several light-harvesting proteins (Lhca and Lhcb) in potato leaves. The tubers of grafted plants showed a 6.5% increase in crude protein, 51% in anthocyanin, and lower carbohydrate content. Goji altered the expression of tubers genes involved in assimilatory sulfate reduction, which subsequently affects cysteine-methionine biosynthesis. Furthermore, the tuber transcriptome shows ABA signaling and transcription factors regulate the expression of key biosynthetic genes involved in inducing the secondary metabolites, such as scopoletin and anthocyanin accumulation, which are primary polyphenols in goji. Our innovative grafting approach offers valuable insights into the interactions between woody and herbaceous plants for developing future strategies to modulate growth efficiency and tuber quality in the face of climate challenges and to meet the demand for nutritious food.

Development of aerial and belowground tubers in potato is governed by photoperiod and epigenetic mechanism.

Plants exhibit diverse developmental plasticity and modulate growth responses under various environmental conditions. Potato (Solanum tuberosum), a modified stem and an important food crop, serves as a substantial portion of the world's subsistence food supply. In the past two decades, crucial molecular signals have been identified that govern the tuberization (potato development) mechanism. Interestingly, microRNA156 overexpression in potato provided the first evidence for induction of profuse aerial stolons and tubers from axillary meristems under short-day (SD) photoperiod. A similar phenotype was noticed for overexpression of epigenetic modifiers-MUTICOPY SUPRESSOR OF IRA1 (StMSI1) or ENAHNCER OF ZESTE 2 (StE[z]2), and knockdown of B-CELL-SPECIFIC MOLONEY MURINE LEUKEMIA VIRUS INTEGRATION SITE 1 (StBMI1). This striking phenotype represents a classic example of modulation of plant architecture and developmental plasticity. Differentiation of a stolon to a tuber or a shoot under in vitro or in vivo conditions symbolizes another example of organ-level plasticity and dual fate acquisition in potato. Stolon-to-tuber transition is governed by SD photoperiod, mobile RNAs/proteins, phytohormones, a plethora of small RNAs and their targets. Recent studies show that polycomb group proteins control microRNA156, phytohormone metabolism/transport/signaling and key tuberization genes through histone modifications to govern tuber development. Our comparative analysis of differentially expressed genes between the overexpression lines of StMSI1, StBEL5 (BEL1-LIKE transcription factor [TF]), and POTATO HOMEOBOX 15 TF revealed more than 1,000 common genes, indicative of a mutual gene regulatory network potentially involved in the formation of aerial and belowground tubers. In this review, in addition to key tuberization factors, we highlight the role of photoperiod and epigenetic mechanism that regulates the development of aerial and belowground tubers in potato.

Organ- and Age-Specific Differences of Dioscorea polystachya Compounds Measured by UPLC-QTOF/MS.

Dioscorea polystachya, named Chinese yam, is widely cultivated as a functional food and natural medicine in China. There is currently little information about the chemical characteristics of Dioscorea polystachya in different organs (tuber cortex and tuber flesh) and at various ages. In this study, an ultra-high-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS) was used to profile chemical compounds in Dioscorea polystachya. As a result, thirty-eight compounds were detected in yam tuber cortex and tuber flesh. More compounds were detected in yam tuber cortex than in tuber flesh. Compounds such as dehydroepiandrosterone, allantoin and flavonoids were selected as biomarker candidates. Dehydroepiandrosterone was found more abundant in tuber flesh, while allantoin and flavonoids showed higher levels in tuber cortex. Furthermore, the levels of dioscin, malvalic acid and sucrose differed significantly among age groups and were highest in the tubers at 2 years. While the levels of allantoin, adenosine and glutamine increased with the growing years and were highest at 4 years. Thus, 2-year old Dioscorea polystachya tubers could be harvested to prepare dioscin, malvalic acid and sucrose supplements. The 4-year-old Dioscorea polystachya tubers would be the best choice for obtaining a large amount of allantoin and adenosine in industrial production.

Agronomical traits, phenolic compounds and antioxidant activity in raw and cooked potato tubers growing under saline conditions.

BACKGROUND: Potato yields and tuber compositions are linked to mechanisms adopted by plants to cope with salinity and often can change after cooking. The current study aimed to evaluate the effects of salinity, variety and cooking method in the composition of potato tubers. Three potato varieties (Spunta, Bellini and Alaska) grown under distinct salt levels (T1: 2.2 ms cm-1 EC, T2: 8.5 ms cm-1 EC before electromagnetic treatment and 6.3 ms cm-1 EC after electromagnetic treatment, T3: 8.5 ms cm-1 EC) were studied. Yield and tuber quality attributes (starch, dry matter, specific density and tuber size) were evaluated. Carotenoids, total and individual phenolics determined by high-performance liquid chromatography (HPLC), relative antioxidant capacity (RAC) and ions content were analyzed, in both raw and water boiled tubers. RESULTS: Tuber yield, starch, dry matter, ions and antioxidants were significantly influenced by the salinity level and variety. The least production and the highest antioxidants were obtained under T3. Antioxidants were influenced by cooking method, the interactions treatment × cooking method and variety × cooking method. Individual phenolic compounds exhibited different response to cooking as quercetin, caffeic acid and catechin decreased significantly after boiling. However, coumaric acid increased in Alaska tubers. CONCLUSION: Salinity level, variety and cooking method are important determinants of tuber yield and composition. Electromagnetic water may be useful to enhance potato production and tuber quality in areas suffering from water salinization. © 2020 Society of Chemical Industry.

Postharvest benzothiazole treatment enhances healing in mechanically damaged sweet potato by activating the phenylpropanoid metabolism.

BACKGROUND: Sweet potato often suffers mechanical damage during harvest, handling, and transportation. Infections, water loss, and quality changes of sweet potato caused by mechanical damage pose great financial losses. Wound healing is an effective method to alleviate such problems. In this study, the effects of postharvest treatment with benzothiazole (BTH) on wound healing of sweet potato was investigated. RESULTS: Postharvest BTH treatment of sweet potatoes promoted lignin accumulation in wounded tissues, and 100 mg L-1 BTH exhibited better effects than 50 mg L-1 or 150 mg L-1 BTH. The biosynthesis of lignin in wounded tissues significantly decreased the weight loss of sweet potatoes. An increase in respiration intensity after BTH treatment was observed. The total phenolic and flavonoid contents and the activity of phenylalanine ammonia-lyase, peroxidase, and polyphenol oxidase were increased in BTH-treated sweet potatoes. This suggests that BTH increases phenylpropanoid metabolism. CONCLUSION: Postharvest 100 mg L-1 BTH treatment could promote wound healing in mechanically damaged sweet potatoes. The activation of the phenylpropanoid metabolism might be the mechanism of action of BTH in wound healing. © 2020 Society of Chemical Industry.

Carbohydrate and bioactive compounds composition of starchy tropical fruits and tubers, in relation to pre and postharvest conditions: A review.

In some tropical countries, people are suffering from both undernourishment and noncommunicable disorders, such as overweight/obesity. Starchy tropical fruits and tubers are of particular interest for their carbohydrate content and for the micronutrients they provide. The present study summarizes the content in carbohydrate, phenolics, carotenoids, and vitamin C, as well as the antioxidant activity of a wide range of tropical fruits and tubers. The energy content of fruits and tubers studied is in the range of 1,200 to 1,800 kJ/100 g of dry weight. They are thus important staple foods and, due to their diversity and seasonality, they can provide energy all year long by alternating the resources in the human diet. Starchy fruit and tuber crops have antiobesity properties as they are bulky, rich in moisture, and contain less than 2% of fat. Noncolored fruit and tubers provide total phenolics at about 20 to 140 mEq/100 g fresh weight. They thus have a high antioxidant capacity, as related to their total phenolic content but also to the presence of carotenoids, such as lutein, mostly in Dioscorea bulbifera and cocoyam. Yellow and orange-fleshed varieties contain more total phenolics and also more provitamin A carotenoids than noncolored fleshed ones. The contents in total phenolic and carotenoid greatly vary with the species and variety. The influence of pre and postharvest conditions on micronutrient content is discussed. Further studies on new processing methods are needed to maximize polyphenols and carotenoids retention in the foods and increase the bioaccessibility of these compounds. PRACTICAL APPLICATION: This paper provides information on the nutritional quality of starchy tropical fruits and tubers. Nutritional quality is studied from the point of view of providing energy and bioactive compounds. The paper aims to promote the use of local resources in tropical areas, which could ultimately limit the adverse effects of food globalization on noncommunicable disorders. It could also lead to tropical countries being less dependent on food imports.

Mobile RNAs and proteins: Prospects in storage organ development of tuber and root crops.

Storage tuber and root crops make up a significant portion of the world's subsistence food supply. Because of their importance in food security, yield enhancement has become a priority. A major focus has been to understand the biology of belowground storage organ development. Considerable insights have been gained studying tuber development in potato. We now know that two mobile signals, a full-length mRNA, StBEL5, and a protein, StSP6A, play pivotal roles in regulating tuber development. Under favorable conditions, these signals move from leaves to a belowground modified stem (stolon) and regulate genes that activate tuberization. Overexpression of StBEL5 or StSP6A increases tuber yield even under non-inductive conditions. The mRNAs of two close homologs of StBEL5, StBEL11 and StBEL29, are also known to be mobile but act as repressors of tuberization. Polypyrimidine tract-binding proteins (PTBs) are RNA-binding proteins that facilitate the movement of these mRNAs. Considering their role in tuberization, it is possible that these mobile signals play a major role in storage root development as well. In this review, we explore the presence of these signals and their relevance in the development and yield potential of several important storage root crops.

Gas chromatography-mass spectrometry metabolomics-based prediction of potato tuber sprouting during long-term storage.

In order to supply potato (Solanum tuberosum L.) tubers for the processed food industry throughout the year, suppliers should provide consistent quality potatoes even after long-term storage. Despite being one of the most important foods, there is no simple way to control tuber quality and, in particular, controlling sprouting. Chemical suppression such as chlorpropham is used to inhibit sprouting, however, the regulatory status of such chemical inhibition differs in each country. Gas chromatography-mass spectrometry-based metabolomics was applied to identify the applicable biomarkers for prediction of potato sprouting during long-term storage. Sprouting was measured in chipping potatoes, and these were also subjected to metabolite profiling to develop a predictive model. The model was based on projections to latent structures (PLS) regression calculated from a metabolome data set obtained before storage and was consistent with actual measured sprouting values. Sucrose, phosphate, and amino acids were selected as valid contributing biomarkers for prediction in a validation field experiment. These biomarkers will contribute to the development of a successful novel method for prediction and control of potato tuber quality during long-term storage.

Effect of citronella essential oil fumigation on sprout suppression and quality of potato tubers during storage.

Effect of citronella essential oil (CEO) fumigation on sprout suppression and quality of potato tubers during storage was investigated. Potato tubers were treated under conditions of single-phase (30 µL L-1, 0-10 d) and dual-phase (30 µL L-1, 0-10 d; 30 µL L-1, 35-90 d) fumigation. Changes in germination rate, weight loss, starch, reducing sugar, gibberellins (GA3), and α-solanine were measured. The results showed that CEO fumigation could control sprouting and improve the quality of potato tubers during storage compared to the non-treated tubers. CEO treatments inhibited the degradation of starch and the increase of reducing sugar content. The production of gibberellins (GA3) was suppressed, and the levels of α-solanine in the skin and flesh of potato tubers were decreased by CEO fumigation. Dual-phase CEO fumigation had a better effect on sprout suppression than single-phase fumigation, and possesses potential for postharvest application.

An integrated metabolomic strategy for the characterization of the effects of Chinese yam and its three active components on septic cardiomyopathy.

Chinese yam (CY), used as both a traditional Chinese medicine and a nutritious food, is an excellent candidate for treating septic cardiomyopathy (SCM). Adenosine, arbutin and allantoin are the major active components in the aqueous extract of CY. The aim of the present study was to interpret the roles of CY, adenosine, arbutin and allantoin in SCM treatment. Firstly, significant physiological indexes were examined to assess the model and treatment effects of CY, adenosine, arbutin and allantoin. Then, a metabolomic approach was utilized to reveal the metabolic disorders in SCM concerning the intervention of CY/adenosine/arbutin/allantoin. The integrated results demonstrated that adenosine, arbutin and allantoin are responsible for the efficacy of CY on SCM treatment by regulating amino acid, arachidonic acid, sphingolipid, glycerophospholipid and glycol metabolism. Moreover, adenosine and/or arbutin could be used as a substitute for CY in treating SCM, and allantoin efficacy was slightly weaker. This integrated metabolomic approach performed excellently in understanding the herbal function and the roles of its components.

Inhibition of plastid PPase and NTT leads to major changes in starch and tuber formation in potato.

The importance of a plastidial soluble inorganic pyrophosphatase (psPPase) and an ATP/ADP translocator (NTT) for starch composition and tuber formation in potato (Solanum tuberosum) was evaluated by individual and simultaneous down-regulation of the corresponding endogenous genes. Starch and amylose content of the transgenic lines were considerably lower, and granule size substantially smaller, with down-regulation of StpsPPase generating the most pronounced effects. Single-gene down-regulation of either StpsPPase or StNTT resulted in increased tuber numbers per plant and higher fresh weight yield. In contrast, when both genes were inhibited simultaneously, some lines developed only a few, small and distorted tubers. Analysis of metabolites revealed altered amounts of sugar intermediates, and a substantial increase in ADP-glucose content of the StpsPPase lines. Increased amounts of intermediates of vitamin C biosynthesis were also observed. This study suggests that hydrolysis of pyrophosphate (PPi) by action of a psPPase is vital for functional starch accumulation in potato tubers and that no additional mechanism for consuming, hydrolysing, or exporting PPi exists in the studied tissue. Additionally, it demonstrates that functional PPi hydrolysis in combination with efficient ATP import is essential for tuber formation and development.

Influence of altitude and enhanced ultraviolet-B radiation on tuber production, seed viability, leaf pigments and morphology in the wild potato species Solanum kurtzianum Bitter & Wittm collected from an elevational gradient.

Climate change could lead to an upward shift in plant distribution, exposing populations to higher levels of ultraviolet (UV)-B radiation. In the framework of an in situ strategy for conserving potato wild relatives, we evaluated the effect of high UV-B levels on natural population of Solanum kurtzianum. The hypothesis is that plants from naturally higher altitudes are more adapted to increased UV-B radiation. Two populations from low and high altitudes were field supplemented using UV-B-lamps (+UV-B) or excluded from it with plastic filters. Additionally, to assess in which extent the plant responses to these artificial experimental conditions are reproducible in natural conditions, three genotypes were cultivated in two mountain experimental gardens (EG) at different elevations. +UV-B treatment induced changes in leaf morphology and increases in phenolic compounds in both populations, indicating plant adaptation, since chlorophylls and reproductive structures were not negatively affected. These results indicate that this environmental factor may not limit the displacement of populations towards sites with higher UV-B levels. Meanwhile, in higher-altitude EG a tubers yield reduction, mainly through a decreased tuber number and a bigger accumulation of phenolic compounds than in +UV-B treatment were observed, suggesting that UV-B is not the only factor involved in plants adaptation to high altitude environments.

Concurrent Overexpression of Arabidopsis thaliana Cystathionine γ-Synthase and Silencing of Endogenous Methionine γ-Lyase Enhance Tuber Methionine Content in Solanum tuberosum.

Potatoes (Solanum tuberosum) are deficient in methionine, an essential amino acid in human and animal diets. Higher methionine levels increase the nutritional quality and promote the typically pleasant aroma associated with baked and fried potatoes. Several attempts have been made to elevate tuber methionine levels by genetic engineering of methionine biosynthesis and catabolism. Overexpressing Arabidopsis thaliana cystathionine γ-synthase (AtCGS) in S. tuberosum up-regulates a rate-limiting step of methionine biosynthesis and increases tuber methionine levels. Alternatively, silencing S. tuberosum methionine γ-lyase (StMGL), which causes decreased degradation of methionine into 2-ketobutyrate, also increases methionine levels. Concurrently enhancing biosynthesis and reducing degradation were predicted to provide further increases in tuber methionine content. Here we report that S. tuberosum cv. Désirée plants with AtCGS overexpression and StMGL silenced by RNA interference are morphologically normal and accumulate higher free methionine levels than either single-transgenic line.

Asada-Halliwell pathway maintains redox status in Dioscorea alata tuber which helps in germination.

Reactive Oxygen Species (ROS) are important regulatory molecules governing physiological processes. In the present study a biochemical and proteome level comparison of two contrasting growth stages of Dioscorea alata tuber namely germinating and mature tuber was performed in order to understand the tuber physiology and biochemistry. Existence of all the component enzymes [APx (ascorbate peroxidase), GR (glutathione reductase), DHAR (dehydroascorbate reductase), MDHAR (mono-dehydroascorbate reductase)] and major products [ascorbate (ASC) and glutathione (GSH)] of the cycle showed an operational Asada-Halliwell cycle in the tuber. A 2.65 fold increase in ASC content & a 3.8 fold increase in GR activity fortified the redox milieu during germination. In contrast a 5 fold higher H2O2 content (due to 3.08 fold lower APx activity) and accumulation of reactive nitrogen species (RNS) such as nitric oxide (NO, 2.4-fold) and S-nitrosothiol (SNO, 2.08 fold) contributed to overall oxidative conditions in the mature tuber. The carbonic anhydrase (CA, 7.5 fold), DHAR (5.31 fold) and MDHAR (7 fold) activities were higher in the germinating tuber in comparison with the mature tuber. GSNO negatively regulated the CA (3.6 & 3.95 fold), MDHAR (7.5 & 1.5 fold) and APx (2.3 & 1.81 fold) while another NO donor, CysNO negatively regulated the DHAR (2.24 & 1.32 fold) activity in the mature and germinating stages respectively indicating again that the lesser inhibition by NO (via nitrosylation) may be because of overall reducing environment in the germinating tuber. Increased SNO leading to S-nitrosylation of dioscorin was confirmed by Biotin switch assay. This is the first report showing dioscorin nitrosylation. The present analysis showed differential redox regulation and also suggests the physiological relevance of CA, DHAR, MDHAR, APx & GR in tuber germination for the first time. These enzymes may be used as potential markers of tuber germination in future.

Transcript profiling reveals that cysteine protease inhibitors are up-regulated in tuber sprouts after extended darkness.

Potato (Solanum tuberosum L.) tubers are an excellent staple food due to its high nutritional value. When the tuber reaches physiological competence, sprouting proceeds accompanied by changes at mRNA and protein levels. Potato tubers become a source of carbon and energy until sprouts are capable of independent growth. Transcript profiling of sprouts grown under continuous light or dark conditions was performed using the TIGR 10K EST Solanaceae microarray. The profiles analyzed show a core of highly expressed transcripts that are associated to the reactivation of growth. Under light conditions, the photosynthetic machinery was fully activated; the highest up-regulation was observed for the Rubisco activase (RCA), the glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and the Photosystem II 22 kDa protein (CP22) genes, among others. On the other hand, sprouts exposed to continuous darkness elongate longer, and after extended darkness, synthesis of chloroplast components was repressed, the expression of proteases was reduced while genes encoding cysteine protease inhibitors (CPIs) and metallocarboxypeptidase inhibitors (MPIs) were strongly induced. Northern blot and RT-PCR analysis confirmed that MPI levels correlated with the length of the dark period; however, CPI expression was strong only after longer periods of darkness, suggesting a feedback loop (regulation mechanism) in response to dark-induced senescence. Prevention of cysteine protease activity in etiolated sprouts exposed to extended darkness could delay senescence until they emerge to light.

Cereal cystatins delay sprouting and nutrient loss in tubers of potato, Solanum tuberosum.

BACKGROUND: Recent studies have reported agronomically useful ectopic effects for recombinant protease inhibitors expressed in leaves of transgenic plants, including improved tolerance to abiotic stress conditions and partial resistance to necrotrophic pathogens. Here we assessed the effects of these proteins on the post-dormancy sprouting of storage organs, using as a model potato tubers expressing cysteine protease inhibitors of the cystatin protein superfamily. RESULTS: Sprout emergence and distribution, soluble proteins, starch and soluble sugars were monitored in tubers of cereal cystatin-expressing clones stored for several months at 4 °C. Cystatin expression had a strong repressing effect on sprout growth, associated with an apparent loss of apical dominance and an increased number of small buds at the skin surface. Soluble protein content remained high for up to 48 weeks in cystatin-expressing tubers compared to control (untransformed) tubers, likely explained by a significant stabilization of the major storage protein patatin, decreased hydrolysis of the endogenous protease inhibitor multicystatin and low cystatin-sensitive cysteine protease activity in tuber tissue. Starch content decreased after several months in cystatin-expressing tubers but remained higher than in control tubers, unlike sucrose showing a slower accumulation in the transgenics. Plantlet emergence, storage protein processing and height of growing plants showed similar time-course patterns for control and transgenic tubers, except for a systematic delay of 2 or 3 d in the latter group likely due to limited sprout size at sowing. CONCLUSIONS: Our data point overall to the onset of metabolic interference effects for cereal cystatins in sprouting potato tubers. They suggest, in practice, the potential of endogenous cysteine proteases as relevant targets for the development of potato varieties with longer storage capabilities.

Polypyrimidine tract-binding proteins of potato mediate tuberization through an interaction with StBEL5 RNA.

Polypyrimidine tract-binding (PTB) proteins are a family of RNA-binding proteins that function in a wide range of RNA metabolic processes by binding to motifs rich in uracils and cytosines. A PTB protein of pumpkin was identified as the core protein of an RNA-protein complex that trafficks RNA. The biological function of the PTB-RNA complex, however, has not been demonstrated. In potato, six PTB proteins have been identified, and two, designated StPTB1 and StPTB6, are similar to the phloem-mobile pumpkin type. RNA binding assays confirmed the interaction of StPTB1 and StPTB6 with discrete pyrimidine-rich sequences of the 3'-untranslated regions of the phloem-mobile mRNA, StBEL5. The promoter of StPTB1 was active in companion cells of phloem in both stem and petioles. Expression of both types was evident in phloem cells of roots and in stolons during tuber formation. RNA accumulation of both PTB proteins was induced by short days in leaves in correlation with enhanced accumulation of StBEL5 RNA. StPTB suppression lines exhibited reduced tuber yields and decreased StBEL5 RNA accumulation, whereas StPTB overexpression lines displayed an increase in tuber production correlated with the enhanced production in stolons of steady-state levels of StBEL5 transcripts and RNA of key tuber identity genes. In StPTB overexpression lines, both the stability and long-distance transport of StBEL5 transcripts were enhanced, whereas in suppression lines stability and transport decreased. Using a transgenic approach, it is shown that the StPTB family of RNA-binding proteins regulate specific stages of development through an interaction with phloem-mobile transcripts of StBEL5.

Next-generation sequencing (NGS) transcriptomes reveal association of multiple genes and pathways contributing to secondary metabolites accumulation in tuberous roots of Aconitum heterophyllum Wall.

MAIN CONCLUSION: The transcriptomes of Aconitum heterophyllum were assembled and characterized for the first time to decipher molecular components contributing to biosynthesis and accumulation of metabolites in tuberous roots. Aconitum heterophyllum Wall., popularly known as Atis, is a high-value medicinal herb of North-Western Himalayas. No information exists as of today on genetic factors contributing to the biosynthesis of secondary metabolites accumulating in tuberous roots, thereby, limiting genetic interventions towards genetic improvement of A. heterophyllum. Illumina paired-end sequencing followed by de novo assembly yielded 75,548 transcripts for root transcriptome and 39,100 transcripts for shoot transcriptome with minimum length of 200 bp. Biological role analysis of root versus shoot transcriptomes assigned 27,596 and 16,604 root transcripts; 12,340 and 9398 shoot transcripts into gene ontology and clusters of orthologous group, respectively. KEGG pathway mapping assigned 37 and 31 transcripts onto starch-sucrose metabolism while 329 and 341 KEGG orthologies associated with transcripts were found to be involved in biosynthesis of various secondary metabolites for root and shoot transcriptomes, respectively. In silico expression profiling of the mevalonate/2-C-methyl-D-erythritol 4-phosphate (non-mevalonate) pathway genes for aconites biosynthesis revealed 4 genes HMGR (3-hydroxy-3-methylglutaryl-CoA reductase), MVK (mevalonate kinase), MVDD (mevalonate diphosphate decarboxylase) and HDS (1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate synthase) with higher expression in root transcriptome compared to shoot transcriptome suggesting their key role in biosynthesis of aconite alkaloids. Five genes, GMPase (geranyl diphosphate mannose pyrophosphorylase), SHAGGY, RBX1 (RING-box protein 1), SRF receptor kinases and ß-amylase, implicated in tuberous root formation in other plant species showed higher levels of expression in tuberous roots compared to shoots. A total of 15,487 transcription factors belonging to bHLH, MYB, bZIP families and 399 ABC transporters which regulate biosynthesis and accumulation of bioactive compounds were identified in root and shoot transcriptomes. The expression of 5 ABC transporters involved in tuberous root development was validated by quantitative PCR analysis. Network connectivity diagrams were drawn for starch-sucrose metabolism and isoquinoline alkaloid biosynthesis associated with tuberous root growth and secondary metabolism, respectively, in root transcriptome of A. heterophyllum. The current endeavor will be of practical importance in planning a suitable genetic intervention strategy for the improvement of A. heterophyllum.

Optimization of in vitro inhibition of HT-29 colon cancer cell cultures by Solanum tuberosum L. extracts.

Secondary metabolites in potato have been reported to possess bioactive properties, including growth inhibition of cancer cells. Because potatoes are widely consumed globally, potential health benefits may have broad application. Thus we investigated growth inhibition of HT-29 colon cancer cell cultures by extracts from 13 diverse genetic breeding clones. Extracts from three pigmented selections (CO97226-2R/R, CO97216-1P/P, CO04058-3RW/RW) inhibited growth of in vitro HT-29 cell cultures more effectively than other clones tested. While inhibition was highest from pigmented selections and pigmented tuber tissue sectors, not all pigmented breeding lines tested had appreciable inhibitory properties. Thus, inhibition was not uniquely linked to pigmentation. Immature tubers had the highest inhibitory properties, and in most cases mature tubers retained very low inhibition properties. Flowers and skins inhibited strongly at lower extract concentrations. An extract consisting of 7.2 mg mL⁻¹ cell culture medium was the lowest effective concentration. While raw tuber extracts inhibited most effectively, a few clones at higher concentrations retained inhibition after cooking. Heated whole tubers retained higher inhibition than heated aqueous extracts. While all aqueous extracts from the two tuber selections (CO97216-1P/P and CO97226-2R/R) inhibited HT-29 cell cultures, inhibition was significantly enhanced in purple pigmented tubers of CO97216-1P/P prepared cryogenically as liquid nitrogen powders compared to extracts from freeze dried samples. Upregulation of caspase-3 protease activity, indicative of apoptosis, was highest among the most inhibitory clone samples. The unique sectorial red pigment expressing selection (CO04058-3RW/RW) provided a model system that isolated expression in pigmented sectors, and thus eliminated developmental, environmental and genetic confounding.