Research Article

Inducing autotetraploids in cassava using oryzalin and colchicine and their in vitro morphophysiological effects

Abstract

Polyploid induction has been used for plant breeding to produce bigger and more robust plants than diploid types. The present study aimed to develop a methodology for in vitro induction of polyploidy in cassava. Apical and lateral microcuttings from the BRS Formosa variety were treated with six oryzalin concentrations for 24 and 48 h. The same methodology was used for colchicine with different concentrations. After 45 days of cultivation and an additional 45 days of subculture, the viability of the explants was assessed and plant acclimatization was performed. Ploidy was determined using flow cytometry. Oryzalin dose and exposure negatively affected cassava explant growth and development compared to untreated explants. Furthermore, apical and lateral explants responded differently to the treatments, showing a diversity in antimitotic sensitivity and effect that is tissue-type specific. In contrast, the doses of 1.25 to 6.25 mM colchicine resulted in high mortality of cassava explants. Therefore, the type of antimitotic affects the morphophysiological behavior of cassava plants in vitro, although apical explants have higher viability and regenerative capacity compared to lateral explants. In addition, the lateral explants have lower mixoploid rates compared to apical explants. Of the 310 plants generated by oryzalin treatments, 277 were diploid, 31 were mixoploid, and 2 were tetraploid. Exposure to oryzalin led to low rate of tetraploids and colchicine caused phytotoxic reactions and death of the explants. The tetraploids were multiplied in vitro to evaluate their yield in the field as well as their behavior against abiotic and biotic stress.

Polyploid induction has been used for plant breeding to produce bigger and more robust plants than diploid types. The present study aimed to develop a methodology for in vitro induction of polyploidy in cassava. Apical and lateral microcuttings from the BRS Formosa variety were treated with six oryzalin concentrations for 24 and 48 h. The same methodology was used for colchicine with different concentrations. After 45 days of cultivation and an additional 45 days of subculture, the viability of the explants was assessed and plant acclimatization was performed. Ploidy was determined using flow cytometry. Oryzalin dose and exposure negatively affected cassava explant growth and development compared to untreated explants. Furthermore, apical and lateral explants responded differently to the treatments, showing a diversity in antimitotic sensitivity and effect that is tissue-type specific. In contrast, the doses of 1.25 to 6.25 mM colchicine resulted in high mortality of cassava explants. Therefore, the type of antimitotic affects the morphophysiological behavior of cassava plants in vitro, although apical explants have higher viability and regenerative capacity compared to lateral explants. In addition, the lateral explants have lower mixoploid rates compared to apical explants. Of the 310 plants generated by oryzalin treatments, 277 were diploid, 31 were mixoploid, and 2 were tetraploid. Exposure to oryzalin led to low rate of tetraploids and colchicine caused phytotoxic reactions and death of the explants. The tetraploids were multiplied in vitro to evaluate their yield in the field as well as their behavior against abiotic and biotic stress.