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Application of Spectrophotometric Fingerprint in Cluster Analysis for Starch Origin Determination

Nikola Sakač1*orcid tiny, Maja Karnaš2orcid tiny, Jasminka Dobša3orcid tiny, Marija Jozanović4*orcid tiny, Vlatka Gvozdić4orcid tiny, Elvira Kovač-Andrić4orcid tiny, Marija Kraševac Sakač4orcid tiny and Bojan Šarkanj5orcid tiny

1University of Zagreb, Faculty of Geotechnical Engineering, Hallerova 7, HR-42000 Varaždin, Croatia

2Josip Juraj Strossmayer University of Osijek, Faculty of Agriculture, Vladimira Preloga 1, 31000 Osijek, Croatia

3University of Zagreb, Faculty of Organization and Informatics, Pavlinska 2, 42000 Varaždin, Croatia

4Josip Juraj Strossmayer University of Osijek, Department of Chemistry, Cara Hadrijana 8/A, 31000 Osijek, Croatia

5University North, Dr Žarko Dolinar Square 1, 43000 Koprivnica, Croatia

Article history:

Received: 7 February 2019

Accepted: 10 March 2020

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Key words:

starch botanical origin, starch-triiodide complex, cluster analysis


The botanical origin of starch is of importance in industrial applications and food processingbecause it may influence the properties of the final product. Current microscopicmethods are time-consuming. Starch consists of an origin-dependent amylose/amylopectinratio. Triiodide ions bind characteristically to the amylose and amylopectin dependingon the botanical origin of the starch. The absorbance of the starch-triiodide complex wasmeasured for wheat, potato, corn, rye, barley, rice, tapioca and unknown origin starch,and within the different cultivars. Each starch sample had specific parameters: starch-triiodidecomplex peak wavelength maximum (λmax/nm), maximum absorbance change atλmaxA) and λmax shift towards the unknown origin starch sample values. The visible absorptionspectra (500-800 nm) for each starch sample were used as a unique fingerprint,and then elaborated by cluster analysis. The cluster analysis managed to distinguish dataof two clusters, a cereal type cluster and a potato/tapioca/rice starch cluster. The cerealsubclusters extensively distinguished wheat/barley/rye starches from corn starches. Datafor cultivars were mostly in good agreement within the same subclaster. The proposedmethod that combines cluster analysis and visible absorbance data for starch-triiodidecomplex was able to distinguish starch of different botanical origins and cultivars withinthe same species. This method is simpler and more convenient than standard time-consumingmethods.

*Corresponding author: +385915830336

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Development of Efficient, Reproducible and Stable Agrobacterium-Mediated Genetic Transformation of Five Potato Cultivars

Allah Bakhsh*orcid tiny

Department of Agricultural Genetic Engineering, Faculty of Agricultural Sciences and Technologies, Nigde Omer Halisdemir University, 51240 Nigde, Turkey

Article history:

Received: 25 December 2018

Accepted: 6 March 2020

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Key words:

potato genetic transformation, genotypes, plant growth regulators, cost-effective protocol


The developments in transformation technology have enabled the scientists to incorporate, mutate or substitute gene(s) leading to a particular trait; advancing it to a point whereonly few technical limitations remain. Genotype dependency and explant types are importantfactors affecting transformation efficiency in potato. In the present study, a rapid, reproducibleand stable Agrobacterium-mediated transformation procedure in potato wasdeveloped by a combination of different plant growth regulators. Leaf discs and internodalexplants of five cultivars of potato, i.e. Lady Olympia, Granola, Agria, Désirée and Innovatorwere infected with Agrobacterium tumefaciens strain LBA4404 containing pBIN19 expressionvector with β-glucuronidase gusA gene under the control of 35S CaMV promoter. Kanamycinwas used as plant selectable marker for screening of primary transformants at concentration of 100 mg/L. Both explants responded positively; internode being more suitable explant forbetter transformation efficiency. Based on GUS histochemical assay, the transformation efficiencywas 22, 20, 18.6, 15 and 10 % using the internodal explant, and 15, 12, 17, 8 and 6 %using leaf discs as explant in Lady Olympia, Granola, Agria, Désirée and Innovator respectively.Furthermore, PCR assays confirmed the presence of gusA and nptII genes in regeneratedplants. The molecular analysis in succeeding progeny showed proper integration andexpression of both genes. The results suggest Lady Olympia as the best cultivar for futuretransformation procedures. Overall, the short duration, rapidity and reproducibility makethis protocol suitable for wider application of transgenic potato plants.

*Corresponding author:

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