Optimization of Hairy Root Induction in <Emphasis Type="Italic">Solanum tuberosum</Emphasis>

An efficient protocol for hairy root induction in Solanum tuberosum was established using Agrobacterium rhizogenes. Transformation of Desiree was more efficient than with Shepody or Purple Majesty. Transformation efficiency of Desiree tuber discs (97% transformation efficiency) was higher in comparison to root (67% transformation efficiency), stem (61% transformation efficiency) and leaf (14% transformation efficiency) explants. Bacterial density, culture parameters, explant type, and genotype influenced the transformation efficiency, as well as the growth of hairy roots. Hairy root transformation efficiency in explants treated with acetosyringone was enhanced from 72 to 89% in Purple Majesty tubers and 56 to 80% in roots during co-cultivation. The growth rate of hairy roots from tuber discs was about 5 times greater than that of hairy roots from leaves. Tuber hairy roots were highly resistant to kanamycin in the absence of NPT II. This study describes the efficient generation of hairy roots in different potato tissues and cultivars that can provide a fast method to generate transformed tissue or be used for production of secondary metabolites.     

Maturity-Adjusted Resistance of Potato (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.) Cultivars to Verticillium Wilt Caused by <Emphasis Type="Italic">Verticillium dahliae</Emphasis>

Verticillium wilt is a fungal disease of potato caused by two species of Verticillium, V. dahliae and V. albo atrum. The pathogen infects the vascular tissue of potato plants through roots, interfering with the transport of water and nutrition, and reducing both the yield and quality of tubers. We have evaluated the reaction of 283 potato clones (274 cultivars and nine breeding selections) to inoculation with V. dahliae under greenhouse conditions. A significant linear correlation (r = 0.4, p < 0.0001) was detected between plant maturity and partial resistance to the pathogen, with late maturing clones being generally more resistant. Maturity-adjusted resistance, that takes into consideration both plant maturity and resistance, was calculated from residuals of the linear regression between the two traits. Even after adjusting for maturity, the difference in the resistance of clones was still highly significant, indicating that a substantial part of resistance cannot be explained by the effect of maturity. The highest maturity-adjusted resistance was found in the cv. Navajo, while the most susceptible clone was the cv. Pungo. We hope that the present abundance of data about the resistance and maturity of 283 clones will help potato breeders to develop cultivars with improved resistance to V. dahliae.     

Rooting Characteristics of <Emphasis Type="Italic">Solanum chacoense</Emphasis> and <Emphasis Type="Italic">Solanum tuberosum in Vitro</Emphasis>

The precise level of environmental control in vitro may aid in identifying genetically superior plant germplasm for rooting characteristics (RC) linked to increased foraging for plant nitrogen (N). The objectives of this research were to determine the phenotypic variation in root morphological responses of 49 Solanum chacoense (chc), 30 Solanum tuberosum Group Phureja – Solanum tuberosum Group Stenotomum (phu-stn), and three Solanum tuberosum (tbr) genotypes to 1.0 and 0.5 N rate in vitro for 28 d, and identify genotypes with superior RC. The 0.5 N significantly increased density of root length, surface area, and tips. All RC were significantly greater in chc than in either phu-stn or tbr. Based upon clustering on root length, surface area, and volume, the cluster with the greatest rooting values consisted of eight chc genotypes that may be utilized to initiate a breeding program to improve RC in potato.     

Isolation and Culture of Pollen Tetrad Protoplasts from <Emphasis Type="Italic">Solanum tuberosum</Emphasis>

Pollen protoplasts provide a sexual and haploid system for haploid production, cell fusion and mutation studies used in plant improvement. Due to the multiploidy, heterozygosity, and often self-incompatibility in tetraploid genotypes, haploid potatoes are desirable for breeding schemes via ploidy manipulations. In this study, two tetraploid varieties and two dihaploid lines of potato were used for pollen tetrad protoplast isolation and culture. The meiotic tetrad buds were first pre-treated at 5 °C for 0–12 days, then the tetrads were transferred into enzyme solutions containing different concentrations of snailase (0.5–1.5%), 0.3 M osmolites (sucrose, mannitol, glucose or sorbitol), 1.0% Cellulose, 0.5% Hemicellulase, 0.5% Pectolyase, 0.3% Sucrose, 3 mM 2-(N-Morpholino) ethane sulfonic acid, 1% polyvinyl pyrrolidone, 0.01% casein hydrolysate and K₃ medium compositions. Among the four donor materials, tetraploid cv. Gannongshu No. 3 (‘GNS No.3’) showed the greatest protoplast yield (74.6 ± 2.4%). In this variety, most of the tetrad protoplasts regenerated a cell wall and continued cell divisions were observed when they were inoculated in K₃ basic medium supplemented with (0.5–1.0) mg/L 2,4-D + (0.1–0.5) mg/L KT + 0.4 mg/L 6-BA +800 mg/L glutamine +100 mg/L serine. ‘GNS No.3’ also showed the greatest first division frequency (21.6 ± 1.5%) and sustained division to form multicellular structures. The study findings suggested that cultured tetrad pollen protoplasts could reverse the gametophytic developmental pattern programmed in vivo to a sporophytic pathway leading to multicellular microspore-derived colonies.     

Predictive Markers for Cold-Induced Sweetening Resistance in Cold Stored Potatoes (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.)

An approach has been developed to screen a large number of potato clones for cold induced sweetening (CIS) resistance in breeding programs. Two key enzymes responsible for reducing sugar accumulation during cold storage were identified. Clones with the A-II isozymes of UDP-glucose pyrophosphorylase coupled with low activity of vacuolar acid invertase enzyme had increased resistance to CIS by forming less suc, which is subsequently hydrolyzed to the undesirable reducing sugars, glc and fru. Six named cultivars and 192 genetically diverse clones from various breeding programs in USA were analyzed over two years for the two key enzymes and sugar concentration in cold stored tubers. The predictability for CIS resistance during cold storage was 94% both years. Clones classified as class A accumulated low concentration of reducing sugar glc during cold storage. It is suggested that these two predictor enzymes can be used for screening parents and selections in potato breeding program.     

Detection of Glycoalkaloids and Chlorophyll in Potatoes (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.) by Hyperspectral Imaging

The purpose of the study was to investigate the use of hyperspectral imaging (HSI) to detect and quantify chlorophyll (Chl) and total glycoalkaloid concentrations (TGA) in potatoes. To create a set of tubers with different concentrations of Chl and TGA, potatoes of four varieties were wounded or treated with red, blue, red/blue, UV-a, UV-b or UV-c light. HSI analyses were performed with a reflection based setup implemented in an industrial potato sorting machine. After hyperspectral analyses, the peel was sampled, and concentrations of Chl and total TGA were determined. Results showed that the HSI system predicted the concentrations of Chl with a relatively high degree of accuracy, and a prediction R² value of 0.92. Prediction of TGA concentrations performed to a much lesser extent, and the overall prediction R² value was only 0.21. Moderate soil covers only affected the prediction powers to a minor degree.     

Ameliorative Effects of Hydrogen Peroxide,Ascorbate and Dehydroascorbate in <Emphasis Type="Italic">Solanum Tuberosum</Emphasis> Infected by Phytoplasma

Phytoplasmal infections cause loss in yield, quality and germination of tubers. Hydrogen peroxide and antioxidants such as ascorbic acid and dehydroascorbate are implicated in signaling against stress. The effects of these chemicals on minituber yield, sprouting and starch content were evaluated in plants testing positive for phytoplasma. Without chemical treatment, positive plants showed significant reductions in leaf pigments, tuber weights and starch contents, compared to uninfected controls, and had more minitubers though fewer sprouted. Hydrogen peroxide and antioxidant treatments of positive plants significantly reduced the number of minitubers, while enhancing their weights and starch contents, and increased the percentage of sprouting minitubers, while leaf pigment content also increased. This research demonstrates potential benefits of hydrogen peroxide and antioxidants in enhancing the yield and quality of tubers not destined for seed in phytoplasma positive plants.     

<Emphasis Type="Italic">In vitro</Emphasis> and <Emphasis Type="Italic">In vivo</Emphasis> Antioxidant Properties of Extracts from <Emphasis Type="Italic">Coptis chinensis</Emphasis> Inflorescence

The inflorescence of cultivated Coptis chinensis has been valued for tea production for many years in China. The antioxidant activities of C. chinensis inflorescence extracts prepared by various solvents were investigated by using several established in vitro systems: 2,2′-azinobis(3-ethylbenzthiazoline-6-sulphonic acid (ABTS), α,α-diphenyl-β-picrylhydrazyl (DPPH) and superoxide radical scavenging assays, reducing power assay, and ferrothiocyanate (FTC) and thiobarbituric acid (TBA) assays. The results showed that the 70% ethanol extract (EE) had the strongest antioxidant activity in vitro among the various extracts. Based on the in vitro results, EE was used to evaluate the antioxidant activity of C. chinensis inflorescence in vivo. The liver and kidney of intoxicated animals showed a significant decrease in superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) levels, while the malondialdehyde (MDA) level showed a significant increase. These changes were significantly reversed after treatment with EE and the standard vitamin E. Thus, the C. chinensis inflorescence may be a valuable natural source that can be applicable to food industries.     

Nitrogen and Phosphorus Use Efficiency in Improved Potato (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.) Cultivars in Southern Ethiopia

Use of nutrient efficient crop species or cultivars within species in combination with improved crop production practices offer the best option for meeting the future food requirements of growing world populations. Efficiency improvement has a key role for nutrient use and it improves both productivity of yield and minimizes risks of losses that potentially harm the environment. To improve nutrient use efficiency (NUE), one strategy is to select cultivars of crops with high NUE. Both nitrogen (N) and phosphorus (P) are among essential macronutrients commonly applied to agricultural crops to optimize yields. Evaluation of potato cultivars with high N and P efficiency is essential for sustainable production of the crop. In Ethiopia, however, little research has been done to elucidate better NUE of potato crop until recently. This study was, thus, designed to evaluate selected potato cultivars (Belete, Gudene, Jalene, Marachere and one local check) for NP use efficiency under low levels of the nutrients in Wolaita Sodo University, Ethiopia during 2014 and 2015. Factorial combination of 3 levels of each N (0, 55.5 and 111 kg N ha^?1 ) and P (0, 19.5 and 39 kg P ha^?1 ) were assigned to the main plots and the 5 cultivars to subplots of split-plot design in three replications. Efficiency indices such as agronomic efficiency, physiological efficiency and recovery efficiency were computed for combined NP nutrients. Results showed that agronomic nitrogen use efficiency, apparent phosphorus use efficiency and phosphorus use efficiencies were significantly influenced by the interaction effect of rates of NP nutrition and variety. Application of NP more than 55.5:19.5 kg ha^?1 will not beneficial due to their less or equal use and recovery efficiencies to 55.5:19.5 kg NP ha^?1. Belete was the most NP efficient variety followed by Jalene and Gudene in all efficiency indices. Therefore, Belete, Jalene and Gudene return better for NP at 55.5:19.5 kg ha^?1 for economical use of fertilizes while obtaining satisfactory yield. It can be concluded that yield response variation of the varieties was related to their differences in NP uptake and use efficiencies; and the improved potato varieties were superior to local check in using NP nutrients efficiently.     

Comparative Study of Lipophilic and Hydrophilic Antioxidants from <Emphasis Type="Italic">In vivo</Emphasis> and <Emphasis Type="Italic">In vitro</Emphasis> Grown <Emphasis Type="Italic">Coriandrum sativum</Emphasis>

Coriander is commonly used for medicinal purposes, food applications, cosmetics and perfumes. Herein, the production of antioxidants in vegetative parts (leaves and stems) of in vivo and in vitro grown samples was compared. In vitro samples were clone A- with notorious purple pigmentation in stems and leaves and clone B- green. Seeds were also studied as they are used to obtain in vivo and in vitro vegetative parts. Lipophilic (tocopherols, carotenoids and chlorophylls) and hydrophilic (sugars, ascorbic acid, phenolics, flavonols and anthocyanins) compounds were quantified. The antioxidant activity was evaluated by radical scavenging activity, reducing power and lipid peroxidation inhibition. The in vivo sample showed the highest antioxidant activity mainly due to its highest levels of hydrophilic compounds. Otherwise, in vitro samples, mainly clone A, gave the highest concentration in lipophilic compounds but a different profile when compared to the in vivo sample. Clones A and B revealed a lack of β-carotene, β- and δ-tocopherols, a decrease in α-tocopherol, and an increase in γ-tocopherol and clorophylls in comparison to the in vivo sample. In vitro culture might be useful to explore the plants potentialities for industrial applications, controlling environmental conditions to produce higher amounts of some bioactive products.     

Complete Genome Analysis of a PVY<Superscript>N-Wi</Superscript> Recombinant Isolate from <Emphasis Type="Italic">Solanum tuberosum</Emphasis> in China

The complete sequence of CF_YL21, a Potato virus Y (PVY) isolate from Solanum tuberosum in China, was determined to be 9718 nucleotides in length, excluding the 3′-terminal poly(A) tail. The viral genome had a single open reading frame of 9186 nucleotides encoding a polyprotein of 3061 amino acids. The polyprotein was predicted to be cleaved into ten functional proteins by three viral proteases. Sequence analyses indicated that CF_YL21 shared 97% nucleotide identity with Wilga5 (PVY^N-Wi), and two putative recombination signals were detected in the P1 and HC-Pro/P3 regions. Phylogenetic analyses, Bayesian Tip-association Significance (BaTS) test, and multiplex RT-PCR assay confirmed that the isolate had the similar molecular and genomic structure with PVY^N-Wi, a PVY strain formed by recombination between PVY^N and PVY^O. To our knowledge, this is the first report of the complete sequence structure of PVY^N-Wi strain from potato in China.     

Yield Response of Potato (<Emphasis Type="Italic">Solanum tuberosum L</Emphasis>.) Genotypes to Late Blight Caused by <Emphasis Type="Italic">Phytophthora infestans</Emphasis> in Uganda

Potato (Solanum tuberosum L.) is a major food and cash crop, mainly grown by small-scale farmers in the highland regions of Uganda. Potato late blight is one of the major diseases limiting production with potential yield losses over 70%, making host resistance a strong element in integrated disease management. This study was carried out to screen and select high yielding potato genotypes with resistance to late blight in Uganda. Forty-eight genotypes, including advanced clones from the population B3C2 of the International Potato Centre, commercial and farmers’ varieties, were evaluated under two environments for two seasons. Trials were laid out in an 8?×?6 alpha lattice design with three replications. Genotypes showed significant differences in yield and resistance to blight. A higher disease severity was observed in Karengyere (56%). The average RAUDPC (= 100 max) across locations indicated that genotypes 395,077.12 and 392,657.8, with disease severity of 12% and 14%, respectively, were the most resistant. Genotypes Victoria (53%) and NKRN59.124 (48%) were the most susceptible. Mean tuber yield under late blight infection was19.8 t ha^?1. The best yielding genotype across sites was 395,112.32 (35.6 t ha^?1) while 394,905.8 (10.3 t ha^?1), yielded the lowest. The mean marketable tuber weight was 8.9 kg with genotypes 395,112.32 and 395,109.34 having the highest marketable weight of 16.5 kg and 15.6 kg respectively. Correlations between yield and yield related parameters were positive (p ≤?0.001), while those between RAUDPC were negative. The following genotypes, 395,112.32, 391,919.3, 393,220.54. 393,077.54, 396,038.107. 392,657.8, Kinigi, 395,014.17, NKRN59.58, NKRK19.17 and 395,011.2, were identified as promising parents for a late blight resistance breeding program. These exhibited high to medium resistance to late blight disease and high yields.     

Comparative Study on the Chemical Composition,Antioxidant Properties and Hypoglycaemic Activities of Two <Emphasis Type="Italic">Capsicum annuum</Emphasis> L. Cultivars (<Emphasis Type="Italic">Acuminatum</Emphasis> small and <Emphasis Type="Italic">Cerasiferum</Emphasis>)

The present study aimed to evaluate for the first time the phenols, flavonoids, carotenoids, capsaicin and dihydrocapsaicin content and the antioxidant and hypoglycemic properties of Capsicum annuum var. acuminatum small and C. annuum var. cerasiferum air-dried fruits. The ethanol extract of C. annuum var. acuminatum small, characterized by the major content of total poliphenols, flavonoids, carotenoids and capsaicinoids, showed the highest radical scavenging activity (IC₅₀ of 152.9 μg/ml). On the contrary, C. annuum var. cerasiferum showed a significant antioxidant activity evaluated by the β-carotene bleaching test (IC₅₀ of 3.1 μg/ml). The lipophilic fraction of both C. annuum var. acuminatum and C. annuum var. cerasiferum exhibited an interesting and selective inhibitory activity against α-amylase (IC₅₀ of 6.9 and 20.1 μg/ml, respectively).     

<Emphasis Type="Italic">In vitro</Emphasis> Antioxidant Activity of Aged Extracts of some Italian <Emphasis Type="Italic">Allium</Emphasis> Species

Antioxidant activity of fresh Allium sativum L. (garlic) is well known and is mainly due to unstable and irritating organosulphur compounds. Fresh garlic extracted over a prolonged period (up to 20 months) produces odourless aged garlic extract (AGE) containing stable and water soluble organosulphur compounds that prevent oxidative damage by scavenging free radicals. The aim of this study was to investigate the in vitro antioxidant activity of aged (up to 20 months) 15% hydroethanolic extracts of different parts (bulbs, bulblets, flower bulblets, flowers, and leaves) of three Allium spontaneous species which are endemic for Italian flora: Allium neapolitanum Cyr., Allium subhirsutum L., Allium roseum L. and to compare it with the in vitro antioxidant activity of aged 15% hydroethanolic extracts of bulbs and leaves of garlic. The antioxidant potential of aged extracts of all species has been evaluated using two different spectrophotometric assays: 2,2-diphenylpicrylhydrazyl (DPPH) test and the ferric reducing/antioxidant power (FRAP) assay. Furthermore the polyphenol content was determined. The aged extracts obtained from the leaves showed the best antioxidant activity, followed by flowers and then by bulbs in both used tests, while flower bulblets and bulblets exhibited lower results or no activity. The polyphenol content was generally directly correlated with antioxidant/antiradical activity. This study confirms the data obtained in previous researches, the wild-type species of Allium and in particular organs other than bulbs are more active and efficacious than garlic bulb. Surely leaves of these Allium spp. deserve special attention.