Regeneration of polyploidy from young thallus segments of Kappaphycus alvarezii was optimized for genetic improvement. Kappaphycus thallus segment cultured on sterile sea water supplemented with various combinations of Indole acetic acid, Kinetin and Acardian Marine Plant Extract Powder revealed differential response on callus proliferation and development of new thallus. Presence of Acardian Marine Plant Extract Powder (3 mg/l) in combination with Indole acetic acid and Kinetin (0.01 mg/l each) had induced the longest emerging thallus. Exposure of thallus to colchicine at 0.01% with above combination was optimal to induce high frequency regeneration of polyploidy mostly from the meristematic cells. Anatomical study of colchicine induced polyploidy revealed larger cortical cells with irregular thickening of epidermal layer. Phase contrast and Scanning Electron Microscopic study revealed increase in cell size in cortical region with significantly larger number of spherical shaped carrageenan globules in colchicine induced polyploidy than normal thallus. Single cells isolated using enzymatic treatments from colchicine induced polyploidy, shown chromosome number with a ploidy status of 4n ≈ 40. Whereas in normal thallus, only half the number of chromosomes (2n ≈ 20) were observed. Polyploidy were successfully acclimatized gradually using raft method for further evaluation. This is the first report reveals the induction and regeneration of polyploidy in Kappaphycus. The possible application of this finding in genetic improvement of Kappaphycus is discussed.
Currently global seaweed industries are facing issue with availability of raw material for processing of carrageenan due to low growth of current planting material. Use of biostimulants in seaweed cultivation is recently paid more attention due to their proven biostimulatory effect, of which, Ascophyllum marine plant extract powder (AMPEP) is a well proven biostimulant to improve the growth and quality of Kappaphycus alvarezii biomass. Hence, 500 kg of AMPEP was purchased and studied its impact on the commercial farming of K. alvarezii from April 2018 to January 2022 in India. Vegetative propagule of K. alvarezii were dipped in an AMPEP with concentration range of: 0.025, 0.05, 0.10, 0.15, 0.20 and 0.25 % for 30, 60, 90 and 120 min. Before out-planting on rafts in shallow coastal water and found that K. alvarezii responded well to a 0.1 % solution with dipping time of 60 min. The percentage of average daily growth rate (ADGR%) of AMPEP-treated plant in a 45 d grow out period was 3.50 ± 0.50 % vs a control of 1.75 ± 0.25 % for the summer and pre-monsoon months (p < 0.05) but no statistically significant differences between the treated and control plants were found during the rainy and winter seasons. Treated plants were found with low incidence of epiphytes, and disease as compared to control plants. The general appearance and health of treated Kappaphycus was good with significant differences in the yield and quality of semi-refined carrageenan (SRC) and dry weed quality (p < 0.05). The cost of AMPEP for producing of additional 1 kg of dry Kappaphycus was 0.066USD. Results of the present study is very encouraging and AMPEP can be used for the production of K. alvarezii biomass for industrial and biorefinery processing as it has been witnessed that there was 16.66 % increase in biomass production in 2021in India.