Review of Related Literature: Multispora Fungus

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Chapter II
REVIEW OF RELATED LITERATURE
This chapter presents the literature gathered from previous
researcher’s work, books and articles online. It basically provides the
information and materials in accordance with evaluation of Antifungal
Potential of White Raddish (Raphanus sativus L.) to Saprolegnia
multispora fungus.
Saprolegnia multispora
Phylum: Heterokontophyta
Class: Oomycota
Order: Saprolegniales
Family: Saprolegniaceae
Genus: Saprolegnia
Species: multispora
Saprolegnia multispora is described from water and floating organic
matter taken in the Tille River, in the Burgundian region of France.
The new species is illustrated and compared with other species of the
genus. Distinguishing characteristics of S. multispora are the
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production of smooth-walled oogonia containing many subcentric
oospores which are (1−) 11–70 (−100) per oogonium. The antheridial
branches supplying the oogonia are predominantly diclinous, but at
times these may be monoclinous and androgynous.
Saprolegniosis is a general term referring to a number of species
of fungi that occur in water and infect the epithelium and eggs of a
variety of fish species including striped bass (Figure 11.9). The
principal species are Saprolegnia parasitica, Aphanomyces sp.,
and Achlya sp. Other species of fungi may occasionally occur, but
nearly all are in the family Saprolegniaceae (Alderman, 1982; Hatai,
1989).
Saprolegnia reproduces either sexually or asexually (Alderman,
1982). Sexual reproduction is by fusion of two gametes to form a
thick-walled oospore, whereas asexual reproduction is by means of
zoospores produced in a zoosporangium, which develops at the end of
non-septate cells. Zoospores are motile by means of flagella after
escaping from the zoosporangia. After going through primary and
secondary stages, these reproductive cells attach to dead or injured
tissue to produce filaments (hyphae) which, in turn, form the visible
mycelium (fungal colony). As the hyphae grow from the dead tissue,
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they invade surrounding healthy tissue and produce necrosis. Once
the fungus becomes established it can kill infected fish. The prognosis
of fungal-infected fish is not encouraging because most antifungal
medicants have similar toxicities to fish and fungi.
Saprolegniosis is identified by the presence of large non-septate,
tubular cells in wet mounts from infected tissues. Saprolegnia
infections are generally external and appear anywhere on the body
surface, fins, or gills. They are often accompanied by pathogenic
bacteria or protozoan parasites, which can also be the predisposing
factor to fungal infection. Fungal colonies, which appear as tufts of
cotton on the fish’s body, may be white, grey, or turn brown as the
mycelium traps mud or silt. Saprolegnia usually does not produce
deep lesions in the muscle, but infected fish swim lethargically and
may linger moribund for several days before death. Aphanomyces sp.
are more likely to produce deep, ulcerated lesions in the muscle of
infected fish (Noga, 1992).
Saprolegnia on incubating eggs can develop rapidly and usually
begins with the fungus attacking injured, infertile, or dead eggs and
then invading the healthy eggs nearby, and eventually totally
engulfing all eggs in a hatching unit. The disease may occur at a wide
range of temperatures with infections more often developing in water
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with below optimum temperature for hatching, but any temperature
shock, either up or down, can be a predisposing factor to fungal
infection on eggs or fish.
The oomycote zoospore morphology and molecular phylogeny
(based on nuclear and mitochondrial genes) confirm these organisms
as members of the stramenopiles and a sister clade, the
photosynthetic Ochrophyta. The oomycote zoospore morphology and
sequence of nucleotides in its rRNA gene confirm these organisms as
stramenopiles (“straw bearers”). Oomycotes are variously called water
molds, white rusts, and downy mildews. Some 50 genera and classes
are recognized. Based on both morphology and molecular data, it is
apparent that the majority of the genera fall into two major clades, the
largely plant-pathogenic peronosporans (which also encompasses the
lagenidialians that are mostly holocarpic animal pathogens such as
Lagenidium and Myzocytiopsis) and the largely saprotrophic free-living
saprolegnians (water molds, including familiar genera such as Achlya,
Dictyuchus, and Saprolegnia as members of the leptomitales, such as
Apodachlya). The white blister rusts (Albugo) are the earliest
branching representative of the peronosporans, which includes the
important plant-pathogenic orders, the pythiales and the
peronosporales. The placement of the rhipidians (such as the sewage
fungus, Sapromyces) is still uncertain because they have been
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associated with both the main clades, depending on the gene being
analyzed. Recent molecular studies have also revealed that a number
of holocarpic oomycote genera of uncertain affiliation, Euryhasma,
Haptoglossa, and Haliphthoros all diverge before the two main clades.
These genera either are exclusively marine or have marine
representatives and are symbiotrophs of seaweeds, nematodes, and
crustaceans, and indicate the marine origins of this lineage.
Like the related hyphochytrids (Pr-14) and true-fungal chytrids
(Pr-35), oomycotes are either saprobes or symbiotrophs. A number of
the more primitive genera such as Eurychasma, Haptoglossa,
Lagenidium, and Myzocytiopsis are holocarpic, completely occupying
their host cells or body cavities. However, most species feed by
extending fungus threads, or hyphae, into invaded tissues, where they
release digestive enzymes and absorb the resulting nutrients. These
hyphae are usually coenocytic, although they may become plugged in
response to injury or to delimit their reproductive organs. Unlike true
fungi, the cell walls use cellulose to form the underlying microfibrillar
skeleton. Although typically considered to be organisms of freshwater
ponds and streams, oomycotes can be found in marine ecosystems,
are widespread in soil, and are symbiotrophs of a wide range of plant
and animal hosts. A few genera even attack vertebrates including fish
(Saprolegnia parasitica) and mammals (Pythium inosidium). Most
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members of the pythians and peronosporans are plant pathogens,
although one genus, Halophytophthora, plays a key saprotrophic role
in breakdown and recycling of disgarded mangrove leaves.
These species are found in lakes, streams, rivers, swamps and
floodplains, many of which are subject to seasonal drying. The
members of the genus Saprolegnia are ubiquitous oomycetes that
form a group of aquatic fungi-like organisms possessing coenocytic
mycelium and cellulose type cell walls. Septation of the mycelium
occurs to delimit reproductive and assimilative portions of the thallus.
These are mostly saprophytes but some are known to be fish
parasites. Unlike most of the eumycetes, the members of this group
remain diploid throughout their life cycles with meiosis occurring in
the gametangia before fertilization. The asexual reproduction is by
biflagellated zoospores formed in the sporangium. Sexual reproduction
is through “gametangial copulation” in which motile male gametes
(sperms) are absent. These are replaced by male nuclei which are
directly injected from the antheridium into the oospheres present in
the oogonium. The character combination of cellulose cell wall,
biflagellated zoospores, gametangial copulation and modern molecular
and biochemical analyses suggest that the oomycetes are closer to
algae (Phaeophyta and Chrysophyta) and higher plants. At present the
oomycetes are no longer considered to be true fungi and are now
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classified amongst “Stramenopiles”, one of the eukaryotic Kingdoms
which includes water molds and brown algae. The position of the
oomycetes as a unique lineage of stramenopile eukaryotes, unrelated
to true fungi but closely related to heterokont (brown) algae, has been
well established using molecular phylogenies that are based on
ribosomal RNA (rRNA) sequences. Surprisingly, botanists of the mid-
1800s had classified Saprolegniaceous organisms within the algal
groups and Saprolegniaceae (Saprolegnia, Achlya, Pythium) was given
an equal status to the algal family of Oedogoniaceae.
The taxonomy of the genus Saprolegnia is mainly based on the
morphological descriptions and the keys provided by Coker, Sparrow,
Seymour and the monumental (and freely available on the internet)
work of Johnson et al. are indispensable. However the morphological
observations are now being supplemented with molecular
characteristics. The polymerase chain reaction coupled with
restriction fragment length polymorphism analysis (PCR-RFLP) has
become a useful tool in taxonomy and is currently used to identify
different species of oomycetes. Amplification of the ribosomal gene is
used for the genetic identification of many organisms because they
comprise both highly conserved sequences during evolution and
highly variable sequences among species and even within species. The
ribosomal nuclear DNA consists of transcribed and non-transcribed
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regions. The ITS1 and ITS2 regions (internal transcribed spacer) are
non-conserved and have been amplified with the PCR method using
universal primers ITS1 and ITS4.
During a survey of zoosporic organisms occurring in the
Burgundian region of France, a new species, Saprolegnia
multispora (F-1247), has been isolated from water samples taken in
the river Tille. The new oomycete is closely related to S. diclina and S.
parasitica as far as its morphological and molecular characteristics
are concerned; however, the oomycetes have their own distinctive
features which separates them from other known oomycetes. S.
multispora was isolated five times from water samples taken at
different places in the Tille river. The isolate F-1247 produces
antheridia and oogonia plentifully, hence it is considered as the type
specimen.
Molecular methods such as DNA fingerprinting, random
amplification of polymorphic DNA (RAPD- PCR), and the polymerase
chain reaction coupled with restriction fragment length polymorphism
analysis (PCR-RFLP) have been effectively applied to taxonomic issues
involving oomycetes. Especially, the use of the internal transcribed
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spacer (ITS) regions of the ribosomal RNA genes (rDNA) in
combination with morphological data has greatly promoted the correct
species identification within oomycetes and other organisms. During a
survey of zoosporic organisms occurring in fish, three Saprolegnia
ferax isolates and a new species, Saprolegnia brachydanis, have been
obtained from fungal-infected zebra fish.
Besides, the mature oospores and the androgynous antheridial
branches cannot be observed in S. brachydanis. Some other
Saprolegnia species such as S. salmonis, S. longicaulis, and S.
multispora also have many differences in the oogonia, oospores, and
antheridial branches from the new species in the study. A BLAST
search with the ITS sequence of the rDNA of S. brachydanis gives
close similarity to these species: S. longicaulis (AY270032) 95.04%, S.
polymorpha (AB219394) 94.9%, S. parasitica (AY455776) 94.6%, S.
salmonis (AY647193) 94.5%, S. australis (AB219385) 94.3%, S.
multispora (AY197329) 93.4%, S. diclina (AY455775) 93.2%.
Along with the investigations on morphology, physiology and
pathogenicity, the ITS sequence data provide valuable information on
'new' or undescribed taxa. Together with morphological differences,
the sequence diversity can support the creation of new species (Paul et
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al., 1999;Paul, 2001;Paulitz et al., 2003;Paul & Steciow, 2004). Some
recent works on the genus Saprolegnia have been carried out in
Argentina (Steciow, 1998(Steciow, 2001(Steciow, 2003Steciow &
Elíades, 2002), but all these works were based on morphological
observations.
A BLAST search yielded the nearest identity (96.8%) with a new
species isolated in France: Saprolegnia multispora ( Paul & Steciow,
2004); other identities were with Achlya oligacantha (80.8%), and
Saprolegnia ferax (79.9%).
Saprolegnia variabilis Steciow & Elíades and S. multispora Paul
& Steciow also possess subcentric oospores type I developing within
smooth, and pitted internal-walled oogonia like those of S. bulbosa
(Steciow & Elíades, 2002;Paul & Steciow, 2004). However, the oogonial
dimensions, number of oospores per oogonia, the nature of antheridial
branches and the morphology of zoosporangia are quite different when
compared with the new species.
Antifungal properties
Antifungal properties a medication that limits or prevents the
growth of yeasts and other fungal organisms. It destroys fungi or
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inhibiting their growth Antifungal properties are exhibited by nutmeg
and it is suggested that nutmeg could be an alternative to the use of
synthetic fungicides to some microorganisms. Nutmeg inhibited the
mycelial growth of Aspergillus niger (Ascomycota), Fusarium
sambucinum (Ascomycota), Pythium sulcatum (Oomycota) and
Rhizopusstolonifer (Zygomycota) (Mvuemba et al., 2009). Nutmeg oil is
very effective in controlling the pathogen Phomopsis azadirachtae
which causes die back in neem (Fathima et al., 2009). Mace oil is
inhibitory to the growth of A. parasiticus and F. moniliforme (Juglal et
al., 2002) and prevented the formation of aflatoxins. Ur-Rahman et al.
(2000) reported the antifungal properties of essential oil from nutmeg.
Phenyl pro- panoids and 8-O-4′ neolignans isolated from mace
exhibited antifungal properties (Zacchino et al., 1997, 1998). Three
lignans, namely, erythro-austrobailignan-6 (EA6), meso-
dihydroguaiaretic acid (MDA) and nectandrin-B (NB), which were
isolated from the methanol extract of nutmeg seed exhibited
antifungal properties. The antifungal property of these lignins against
Alternaria alternate, Colletotrichum coccodes, C. gloeosporioides,
Magnaporthe grisea, Agrobacterium tumefaciens, Acidovorax konjaci
and Burkholderia glumae were studied in vitro and in vivo. The fungi
were sensitive to the three lignins and showed varied antimicrobial
activity. In in vivo studies it was reported that the various lignans
suppressed the development of rice blast, wheat leaf rust, barley
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powdery mildew, tomato late blight and rice sheath blight (Cho et al.,
2007).
Peptides with antibacterial and antifungal properties are
present in skin secretions of nonmammalian vertebrates that include
Agnatha (hagfish and lampreys), Elasmobranchii (sharks and rays),
Teleostei (bony fish), and Amphibia and constitute a component of the
organism’s system of innate immunity. Host-defense peptides produce
cell death either by disrupting the integrity of the cell membrane or by
translocating across the membrane without producing lysis and
binding to intracellular targets. In the first category, there has been
little or no evolutionary pressure to conserve amino-acid
sequences that define antimicrobial activity but, in the great majority
of cases, the peptides are cationic, contain a relatively high proportion
of hydrophobic amino acids, and one or more domains with the ability
to adopt an amphipathic α-helical conformation on binding to the cell
membrane. The second category, of particular importance in the host-
defense systems of teleost fish, includes strongly cationic peptides
derived from the proteolytic cleavage of histones, particularly histones
H1, 2A, and 2B, and ribosomal proteins.
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The first description of antifungal properties in azole
compounds was in 1944 when researchers studying biotin deficiency
in animals noted that benzimidazole, which is structurally similar to
biotin, had activity against yeasts. This finding was not followed up at
the time, but later groups routinely screened other azoles for
antifungal activity. Early agents to be developed in the group,
including clotrimazole, ketoconazole, and miconazole, are generally
now confined to topical use because of unfavorable safety profiles.
Further refinements and chemical modification of these
compounds resulted in the triazole group, which is characterized by a
five-member azole ring. Triazoles such as fluconazole, itraconazole,
voriconazole, and posaconazole are widely used for prophylaxis and
treatment of invasive fungal disease (IFD). The azoles share a common
mode of action: inhibition of the fungal cytochrome P450 enzyme
(CYP450) lanosterol 14-α-demethylase, which facilitates conversion of
lanosterol to ergosterol, a component of the fungal, but not the
human, cell membrane. Disruption of ergosterol biosynthesis causes
damage to the fungal cell membrane, accumulation of methylsterols,
and either cell lysis leading to fungal cell death or inhibition of fungal
cell growth, depending on the azole and fungal species.
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Radish (Raphanus sativus L.)
Kingdom: Plantae
Class: Magnoliopsida
Order: Brassicales
Family: Brassicaceae
Genus: Raphanus
Species: sativus L.
Radish is chock full of dietary fibre, which helps with digestion issues.
If you have enough mooli salad every day, your bowel movement will be
smooth. What's more, you won't have any constipation with enough intake of
radishes. While there is no cure for the common cold, and the cough that comes
with it during winters, radishes can actually help you fight these ailments. This
root veggie has anti-congestive properties, which help clear the mucus from
your throat and respiratory tract. So, instead of reaching for a decongestant
that'll make you sleepy, indulge in some mooli. Full of vitamin A, C, E, B6,
potassium, and other minerals, radishes can give your whole body an immunity
boost. Mooli is also high on antioxidants and anthocyanins, which means that
it's quite good for your heart as well. Of course, you have to have this veggie
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regularly to gain benefits in the long term. Rich in potassium, radishes can help
keep blood pressure under control by maintaining the sodium-potassium
balance in the body. This anti-hypertensive property is the reason why
everybody should have enough mooli in winters, when high blood pressure
issues get worse, if not taken care of. Apart from all the vitamins in radishes,
they also have phosphorous and zinc. These, when combined, can help get rid
of dryness, acne and rashes. What's more, radishes also have a high water
content, and keep the body naturally hydrated. Have enough mooli, and you'll
have glowing skin all winter (Health Magazine, 2017).
A basic 9-kD protein was purified from seeds of radish
(Raphanus sativus L.). The 43 amino-terminal amino acids show
extensive sequence identity with nonspecific lipid transfer proteins
from other plant species. The radish seed nonspecific lipid transfer
protein-like protein inhibits the growth of several fungi in vitro.
Related Foreign Study
Study 1
Antifungal activity of radish, Raphanus sativus, Family brassicaceae
(Cruciferae) by Marie P. Nakamura
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ABSTRACT
Medicinal plants and herb plants contain substances known to
modern and ancient civilization for their healing properties. They are
important sources of effective and affordable drugs. One of these
medicinal plants includes radish (Raphanus sativus) which gives
medical and economic importance. The antifungal property of
Raphanus sativus will be a great interest in the field of research
because it can be useful for researchers on drug industries to develop
new antifungal drugs that is safe and effective for public use. Of
particular interest in the field of Medical Technology is the importance
of antifungal susceptibility testing on the threshold of each fungal
species in the different antifungal drugs.
The experimental method were employed in the study. The data
were measured using the chi-square to determine the significant
difference in the amount of extract yield according to solvent used.
Statistical analysis showed that difference in the value of three solvent
used in the soxhlet method and reflux distillation method was not
statistically significant. The study showed that yield of extract using
method was significantly higher than reflux distillation method.
Soxhlet was found to be more efficient extraction process. The study
showed that radish exhibited antifungal activity against four fungal
strains. The most superior extract used in inhibiting the fungi was the
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ethanol extract. The minimum inhibitory concentration of the extracts
had a range of 10-20 mm against fungal test organisms.
There were four possible bioactive agents present in the extracts
based on the results in the Thin Layer Chromatography. It is probable
that these agents are made up of steroids, alkaloids, isothiocyanate,
glucosides and anthocyanin.
The study concluded that the radish extracts exhibited
antifungal activity against four fungal starins. Ethanol was found to
be the best solvent used in obtaining the maximum yield of bioactive
substances in the extract. The possible bioactive agents present in the
different extract may vary in their chemical structures.
It is recommended that further study should be done on the
production of low-cost antifungal ointment and systemic drugs using
radish extract. Optimization experiments should be made on the
extraction and purification of the radish extract. Comparative in-depth
study on the antifungal activity of radish should be performed.
Beating a Candida overgrowth is about much more than simply
taking supplements – you need to choose the right diet too. Antifungal
foods can play an important role in fighting off fungal pathogens like
Candida, and I’ve created a list of the best anti-Candida foods that
you can use just for this purpose.
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Foods like garlic, rutabaga and coconut oil are natural
antifungals that you probably already know. But today I’m going to
add another antifungal food to the list. Delicious in salads or stir-fries,
this crunchy root vegetable packs a powerful punch when it comes to
fighting off fungal pathogens. I’m talking, of course, about the humble
radish.
Radish as An Antifungal; There has been an increasing amount
of research into radish as an antifungal, particularly over the last 10
years or so. This is a result of the deepening crisis in emergency
wards, where fungal pathogens like Candida can have life-threatening
implications for those with depleted immune systems. The universe of
antifungal drugs is much smaller than that of antibiotics, so
researchers are constantly looking for promising new antifungal
compounds.
Some of this research has focused on a plant defensin known as
RsAFP2, which is found in radishes. Defensins are simply proteins
found in animals and plants that have antimicrobial qualities. Plants
are regularly threatened by fungal pathogens, just like we are, and the
antifungals that they use to protect themselves often work for us too.
According to a Belgian research team in 2009, “antifungal
protein 2 (RsAFP2) from radish induces apoptosis [cell death] … in the
human pathogen Candida Albicans.” Another research team in 2012
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discovered the reason for this – RsAFP2 interacts with the Candida
Albicans cell wall, damaging it and effectively destroying the cell.
It has also been shown that this particular antifungal protein
acts to prevent Candida’s transition from its yeast form to its more
virulent fungal form. This is a very useful property, and one of the
reasons that Candida albicans is just a difficult opponent. When
Candida morphs into its fungal form it grows hyphae which enable it
to spread along your digestive tract much more rapidly. Any
compound that prevents it from doing so will help to slow down the
spread of a Candida overgrowth.
Separately, researchers have also managed to identify two
different antifungal proteins in radish seeds. The seeds need these
antifungals to protect themselves during the time before germination,
and during the early part of their growth when they are particularly
vulnerable to microbial infection. Research is continuing on both the
antifungal protein RsAFP2 and those antifungals found in the radish
seeds. Here are a few of the most recent studies:
In Vitro Activity of the Antifungal Plant Defensin RsAFP2
against Candida Isolates and Its In Vivo Efficacy in Prophylactic
Murine Models of Candidiasis (Tavares et al, 2008)
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The antifungal plant defensin RsAFP2 from radish induces
apoptosis in a metacaspase independent way in Candida Albicans
(Aerts et al, 2009)
The plant defensin RsAFP2 induces cell wall stress, septin
mislocalization and accumulation of ceramides in Candida Albicans
(Thevissen et al, 2012)
Radish seeds have previously been shown to contain two
homologous, 5-kD cysteine-rich proteins designated Raphanus
sativus-antifungal protein 1 (Rs-AFP1) and Rs-AFP2, both of which
exhibit potent antifungal activity in vitro. We now demonstrate that
these proteins are located in the cell wall and occur predominantly in
the outer cell layers lining different seed organs. Moreover, Rs-AFPs
are preferentially released during seed germination after disruption of
the seed coat. The amount of released proteins is sufficient to create a
microenvironment around the seed in which fungal growth is
suppressed. Both the cDNAs and the intron-containing genomic
regions encoding the Rs-AFP preproteins were cloned. Transcripts
(0.55 kb) hybridizing with an Rs-AFP1 cDNA-derived probe were
present in near-mature and mature seeds. Such transcripts as well as
the corresponding proteins were barely detectable in healthy
uninfected leaves but accumulated systemically at high levels after
localized fungal infection. The induced leaf proteins (designated Rs-
AFP3 and Rs-AFP4) were purified and shown to be homologous to
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seed Rs-AFPs and to exert similar antifungal activity in vitro. A
chimeric Rs-AFP2 gene under the control of the constitutive
cauliflower mosaic virus 35S promoter conferred enhanced resistance
to the foliar pathogen Alternaria longipes in transgenic tobacco. The
term "plant defensins" is proposed to denote these defense-related
proteins (Terras FR, et al. Plant Cell. 1995).
Study 1
SCREENING IN-VITRO ANTIFUNGAL ACTIVITY OF RAPHANUS
SATIVUS L. VAR. CAUDATUS by Afshan Siddiq and Ishrat Younus,
2015.
ABSTRACT
The aim of present investigation was to evaluate antifungal
activity of ethanolic extract of Raphanus sativus L. var. caudatus. The
antifungal activity using agar disk diffusion method was determined
against six fungal strains; Aspergillus niger, Trichphyton rubrum,
Microsporum canis, Fusarium lini, Candida glabrata and Candida
albicans. Four different concentrations (50, 100, 200 and 400μg/ml)
of Raphanus caudatus extract and standard drug (Miconazole) were
tested against fungi and zone of growth inhibition were measured.
Raphanus caudatus exhibited remarkable antifungal activity (p <
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0.05) against all fungi at all tested concentrations except Fusarium
lini which was inhibited only at higher concentrations (400 and
800μg/ml). It is noted that with increase in concentration, the zones
of growth inhibition were also increased. Thus Raphanus caudatus
could be a lead for development of antifungal agents in future.
Phytochemical Screening
According to Rueras (2017), cited from Ding (2017),
Phytochemicals is defined as non-nutritive plant chemicals that have
protective or disease preventive properties. She also stated that
phytochemicals is the natural chemical compounds found in plants
that make up its color. In addition, Cadapan and Alinsugay (2018)
stated that phytochemicals are non-essential nutrients, meaning, the
human body or sustaining life does not require them. Moreover,
Rueras (2017) also stated in her recent study that Phytochemical
Screening is the extraction, screening and identidfication of medically
active substance found in plants. She then added that there are some
bioactive substances that can be derived from plants such as
flavanoids, alkaloids, carotenoids, tannin, antioxidants and phenolic
compounds.
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Alkaloids
According to Cadapan and Alinssugay (2018), alkaloids are
defined as basic alka-like nitrogen containing organic constituents
that occur mainly in plants. They also added that nitrogen in alkaloid
molecule is derived from amino acids, often with heterocyclically bond
nitrogen. Rueras (2017) also stated that alkaloid-producing tissue
cultures provide an alternative for exploiting slow growing or protected
plant. She then added that alkaloids can be utilized for biosynthetic
studies and for commercial producton of valuable alkaloids.
Anthocyanosides
Cadapan and Alinsugay (2018) states that antocyanosides are
specific chemical elements in plants that control pigmentation known
as super-antioxidants. They also added that anthocyanosides are
highly prized as health boosting substances because of their
antioxidant properties.
Flavanoids
Robertson (2014) states that flavanoids are group of plant
metabolites thought to provide health benefits through cell signalling
pathways and antioxidant effects. He also stated that these molecules
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are found in a variety of fruits and vegetables. Rueras (2017) also
stated that flavanoids are important for growth and development in
plant, attraction of pollinator animals, nitrogen-fixation in leguminous
plants, and for protection against damage by herbivores, microbes,
UV and reactive oxygen species, Furthermore,
flavanoids are associated with skin protection, brain function, blood
sugar and blood pressure regulation, in addition to antioxidant and
anti-inflamatory activity.
Glycosides
Rueras (2017) cited from Kumar (2016), asserted that glycosides
are natural substances that contain carbohydrates where glycosidic
part of the molecule is connected with organic radiacal that is not
sugar.
Phenolic Compounds
Rueras (2017), cited from Cheynier (2013) stated that phenolic
compounds are a large class of plant secondary metabolites, showing
a diversity of structures, from rather simple structures. She also
added that phenolic compounds are important for the quality of plant
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based foods for they are responsible for the color of red fruits, juices
and wines.
Saponins
Rueras (2017) cited from Nwokocha (2017), defined saponins as
a naturally occuring surface active glycosides produced by plants,
lower marine animals, and some bacteria. She then added that
saponins are phytochemical that produce foam when dissolved in
water. Furthermore, Cadapan and Alinsugay (2018) stated that
saponins consist of a polycyclic aglycones attached to anyone or more
sugar side chains. They also added that it has a bitter taste and some
are toxic known as sapotoxin.
Tannins
Cadapan and Alinsugay (2018) asserted that tannin, also called
tannic acid, is a group of pale-yelow to light brown amorphous
substances in the form of powder, flakes, or a spongy mass, widely
distributed in plants. They also added that tannin solution are acid
and have an astringent taste. Rueras (2017), cited from Constabel
amd Barbehemm (2011), mentioned that tannins are most abundant
secondary metabolites made by plants, commonly ranging from 5% to
10% dry weight of tree leaves. She also added that tannins have no
30
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effect on protein digestion in insect herbivores contatry to early
theories. Moreover,Rueras asserted that tannin structure have
important effect in biochemical activity.
31
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CONCEPTUAL FRAMEWORK
Evaluation of Antifungal Potential of White Radish
(Raphanus sativus L.) to Saprolegnia multispora
Fungus
Independent Variable Dependent Variable
Antifungal Potential of
Saprolegnia multispora
White Radish
Fungus
(Raphanus sativus L.)
Figure 1. This figure shows the independent and dependent variable.
The independent variable is Antifungal Potential of White Radish
(Raphanus sativus L. and the dependent variable Saprolegnia
multispora Fungus.
32

Review of Related Literature: Multispora Fungus

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Senior High School

POLOMOLOK NATIONAL HIGH SCHOOL

REVIEW OF RELATED LITERATURE

This chapter presents the literature gathered from previous

researcher’s work, books and articles online. It basically provides the

information and materials in accordance with evaluation of Antifungal

Potential of White Raddish (Raphanus sativus L.) to Saprolegnia

Saprolegnia multispora is described from water and floating organic

matter taken in the Tille River, in the Burgundian region of France.

genus. Distinguishing characteristics of S. multispora are the

production of smooth-walled oogonia containing many subcentric

branches supplying the oogonia are predominantly diclinous, but at

times these may be monoclinous and androgynous.

Saprolegniosis is a general term referring to a number of species

variety of fish species including striped bass (Figure 11.9). The

principal species are Saprolegnia parasitica, Aphanomyces sp.,

and Achlya sp. Other species of fungi may occasionally occur, but

nearly all are in the family Saprolegniaceae (Alderman, 1982; Hatai,

Saprolegnia reproduces either sexually or asexually (Alderman,

1982). Sexual reproduction is by fusion of two gametes to form a

thick-walled oospore, whereas asexual reproduction is by means of

zoospores produced in a zoosporangium, which develops at the end of

non-septate cells. Zoospores are motile by means of flagella after

escaping from the zoosporangia. After going through primary and

secondary stages, these reproductive cells attach to dead or injured

tissue to produce filaments (hyphae) which, in turn, form the visible

they invade surrounding healthy tissue and produce necrosis. Once

of fungal-infected fish is not encouraging because most antifungal

medicants have similar toxicities to fish and fungi.

Saprolegniosis is identified by the presence of large non-septate,

tubular cells in wet mounts from infected tissues. Saprolegnia

infections are generally external and appear anywhere on the body

surface, fins, or gills. They are often accompanied by pathogenic

bacteria or protozoan parasites, which can also be the predisposing

factor to fungal infection. Fungal colonies, which appear as tufts of

mycelium traps mud or silt. Saprolegnia usually does not produce

are more likely to produce deep, ulcerated lesions in the muscle of

infected fish (Noga, 1992).

Saprolegnia on incubating eggs can develop rapidly and usually

then invading the healthy eggs nearby, and eventually totally

range of temperatures with infections more often developing in water

with below optimum temperature for hatching, but any temperature

shock, either up or down, can be a predisposing factor to fungal

infection on eggs or fish.

The oomycote zoospore morphology and molecular phylogeny

(based on nuclear and mitochondrial genes) confirm these organisms

as members of the stramenopiles and a sister clade, the

photosynthetic Ochrophyta. The oomycote zoospore morphology and

sequence of nucleotides in its rRNA gene confirm these organisms as

stramenopiles (“straw bearers”). Oomycotes are variously called water

are recognized. Based on both morphology and molecular data, it is

largely plant-pathogenic peronosporans (which also encompasses the

lagenidialians that are mostly holocarpic animal pathogens such as

Lagenidium and Myzocytiopsis) and the largely saprotrophic free-living

saprolegnians (water molds, including familiar genera such as Achlya,

Dictyuchus, and Saprolegnia as members of the leptomitales, such as

Apodachlya). The white blister rusts (Albugo) are the earliest

branching representative of the peronosporans, which includes the

important plant-pathogenic orders, the pythiales and the

peronosporales. The placement of the rhipidians (such as the sewage

fungus, Sapromyces) is still uncertain because they have been

analyzed. Recent molecular studies have also revealed that a number

of holocarpic oomycote genera of uncertain affiliation, Euryhasma,