Polyploid induction is of utmost importance in horticultural plants for the development of new varieties with desirable morphological and physiological traits. Polyploidy may occur naturally due to the formation of unreduced gametes or can be artificially induced by doubling the number of chromosomes in somatic cells. In this experiment, a protocol for in vitro polyploid induction of highbush blueberry (Vaccinium corymbosum L.) leaf tissues was studied by using different concentrations of colchicine and oryzalin. Oryzalin was found to be highly toxic to this species, while the adventitious shoot organogenesis media enriched with 25 and 250 µM colchicine was able to induce polyploidization, with significant differences among the treatments used. Higher concentrations of both antimitotic agents led to the browning and death of the leaf tissues. The polyploids obtained showed several morphological differences when compared with the diploid shoots. Flow cytometry analysis was used to confirm the ploidy level of the regenerated shoots, demonstrating that a total of 15 tetraploids and 34 mixoploids were obtained. The stomatal sizes (length and width) of the tetraploids were larger than those of the diploids, but a reduced stomatal density was observed as compared to the controls. These shoots will be acclimatized and grown until they reach the reproductive phase in order to test their potential appeal as new varieties or their use for breeding and genetic improvement.
Fluorescent lights are typically used as light source in indoor horticultural production, including micropropagation. However, light emitting diodes (LEDs) have been recently used for plant growth under controlled environment. Major advantage of LEDs is wavelength specificity, that allows to adequately adjust the spectra according to plant needs. The possibility of using LED as primary light source for the micropropagation of Pyrus communis L. has been investigated in this work. It was proceeded to the optimization of a protocol of micropropagation using a LED lamp as the primary light source inside of the growth chamber with specific wavelengths, to reduce energy consumption and increase the quality of the micro propagated plants. Explants were maintained in a growth chamber and exposed to three different continuous spectrum LED lamps (AP67, NS1, G2) as a primary light source and fluorescent lamps (control) for 4 weeks. At the end of four weeks period, it was proceeded to the morphometric and biochemical analysis. Shoot and leaf growths were more influenced by LED lamps as compared to fluorescent lamps (control) in both cultivars. The results showed that the shoots of William and San Giovanni cultivars showed significant differences in morphological and physiological traits, as well as in chlorophyll, carotenoid, and MDA contents. Highest number of neo-formed shoots and nodes were observed in the plantlets of cv William under AP67 LED followed by NS1 and G2 LED lights respectively as compared to the white light (control), whereas same trend was observed in cv San Giovanni under AP67 LED, but it showed higher shoots and node numbers under control LED lamps as compared to both NS1 and G2. The photosynthetic pigments were significantly decreased in leaves of both cultivars when grown under LEDs as compared to the control fluorescent lamps. Moreover, the AP67 LED light had also significant effects on the protein and MDA contents in the leaves of both cultivars in comparison to all other treatments. This work underlines the importance of the modulation of light sources in relation with different species and varieties, allowing optimizing the proliferation phase and the efficiency of Pyrus communis L. micropropagation protocol. Moreover, this protocol can be improved with further studies to examine the response of the plantlets to the ex-vitro acclimatization because the possibility of using LED light for the micropropagation of pear can be considered as a valuable alternative for its ecologically sustainable production.
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