GMU:Experimental Microbiology/Haematococcus pluvialis

Haematococcus pluvialis Introduction “Green microalgae” comprise more than 7000 species growing in a variety of habitats. Haematococcus pluvialis (Chlorophyceae, Volvocales) is unicellular fresh water microalga distributed in many habitats worldwide. The freshwater unicellular biflagellate green microalgae H. pluvialis Flotow belongs to the class Chlorophyceae, order Volvocales and family Haematococcaseae (Bold and Wynne, 1985; Eom et al., 2006). It is also known as Haematococcus lacustris or Sphaerella lacustris. Haematococcus was first described by J. Von Flotow in 1844 and later in 1899 Tracy Elliot Hazen extensively presented its biology and life cycle (Hazen, 1899; Leonardi et al., 2011).

Physiology Cellular structure of H. pluvialis is similar to most of other members of volvocalean unicellular green algae. The life cycle of H. pluvialisconsists of four types of distinguishable cellular morphologies: macrozooids (zoospores), microzooids, palmella, and hematocysts (aplanospores) (Hazen, 1899; Elliot, 1934). Macrozooids (zoospores), microzooids, and palmella stages are usually called “green vegetative phase” .Hematocysts (aplanospores) are referred as “red nonmotile astaxanthin accumulated encysted phase” of the H. pluvialis life cycle). Macrozooids (zoospores) are spherical, ellipsoidal, or pear-shaped cells with two flagella of equal length emerging from anterior end, and a cup-shaped chloroplast with numerous, scattered pyrenoids The macrozooid cells are between 8 and 20 μm long with a distinct gelatinous extracellular matrix of variable thickness. Numerous contractile vacuoles are irregularly distributed near the protoplast surface of the cell (Hagen et al., 2002). These flagellated fast-growing vegetative cells predominate under favorable culture conditions in the early vegetative growth stage .

Reproduction Macrozooids may divide into 2–32 daughter cells by mitosis (Wayama et al., 2013) .Under unfavorable environmental or culture conditions, macrozooids start losing flagella, and expand their cell size. They form an amorphous multilayered structure in the inner regions of the extracellular matrix or the primary cell wall as they develop into non-motile “palmella” and become resting vegetative cells (Hagen et al., 2002) .With the continued environmental stress (i.e., nutrient deprivation, high light irradiance, high salinity) and cessation of cell division, “palmella” transform into the asexual “aplanospores” At this stage, cells contain two distinct structures, a thick and rigid trilaminar sheath, and secondary cell wall of acetolysis-resistant material. Such cells become resistant to prevailing extreme environmental conditions (Santos and Mesquita, 1984; Boussiba and Vonshak, 1991). Mature aplanospores; accumulate large amounts of secondary carotenoids, particularly astaxanthin, in lipid droplets deposited in the cytoplasm, which results in a characteristic bright red color of these cells (Hagen et al., 2002). Some H. pluvialis strains are reported to be capable of accumulating astaxanthin without forming aplanospores (Brinda et al., 2004). Once environmental or culture conditions return to optimal, red aplanospores germinate to form flagellated zoospores to initiate a new vegetative growth cycle gametocysts. Sexual reproduction is rarely observed in H. pluvialis, and has been largely replaced by vegetative reproduction (Triki et al., 1997).

In some cases, gametogenesis may occur in aplanospores .Such process requires an exposure to extreme adverse conditions (freezing, desiccation, or nutrient starvation) followed by return to favorable culture conditions. During gametogenesis, aplanospore cells can produce up to 64 gametes which are known as microzooids. The microzooids are smaller in size (< 10 μm) compared to the zoospores (20–50 μm), and exhibit high-speed motility after their release from Habitat H. pluvialis is common in small transient freshwater bodies and widely distributed in many habitats worldwide. It occurs primarily in temporary water bodies like ephemeral rain pools, artificial pools, natural and man-made ponds, and birdbaths (Czygan, 1970; Burchardt et al., 2006). This microalga can be usually found in temperate regions around the world and has been isolated from Europe, Africa, North America, and Himachal Pradeslv India (Pringsheim, 1966; Suseela and Toppo, 2006). It has been also found across diverse environmental and climate conditions: in brackish water on the rocks on the seashore (Chekanov et al., 2014); freshwater basin in the rock filled with melted snow on Blomstrandhalvøya Island (Norway) (Klochkova et al., 2013); dried fountain near Rozhen, Blagoevgrad in Bulgaria Gacheva et al., 2015, freshwater fishpond in Bihor, Romania (Dragos et al., 2010); rooftop surface of a building of KIOST in Seoul Korea (Kim et al., 2015). It is well suited for survival under conditions of expeditious and extreme in light, temperature, and salt concentration that would be deleterious to many other microalgae, due to its ability to encyst (become enclosed by thick membrane) in a rapid manner (Proctor, 1957). Commercial Uses Many species of microalgae have been used as source of nutrient rich food, feed, and health promoting compounds. Among the commercially important microalgae, Haematococcus pluvialis is the richest source of natural astaxanthin which is considered as “super anti-oxidant.” Natural astaxanthin produced by H. pluvialis has significantly greater antioxidant capacity than the synthetic one. Astaxanthin has important applications in the nutraceuticals, cosmetics, food, and aquaculture industries. It is now evident that, astaxanthin can significantly reduce free radicals and oxidative stress and help human body maintain a healthy state. With extraordinary potency and increase in demand, astaxanthin is one of the high-value microalgal products of the future.