Querctin from endofungi

1. Isolation and extract of “querctin “from endophtic
fungi
Historical introduction of isolation of “querctin”from plant
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1
-Definition of endophtic fungi
2
-Techniques for isolation and detection of fungal endophytes
3
-Economical and Medical importance of “querctin”
4
-The ability and Strategies of extract “querctin “ from endophtic fungi
5

2. 1- Historical introduction of isolation of “querctin”from plant
Quercetin, a plant pigment is a potent antioxidant flavonoid and more specifically a flavonol,
found mostly in onions, grapes, berries, cherries, broccoli, and citrus fruits. It is a versatile
antioxidant known to possess protective abilities against tissue injury induced by various
drug toxicities.
The flavonoid quercetin was isolated effectively from the leaves of Fenugreek (Trigonella
foenum-graecum) is an annual herb that belongs to the family Leguminosae commonly
grown in India, Pakistan, and some Middle Eastern countries, which has many beneficial
medicinal effects
Both the leaves and seeds of the fenugreek plants were widely consumed as food and
medicine in Indo-Pak subcontinent and also in other countries. Fenugreek is rich in the
source of vitamins, iron, β-carotene, etc
It also has been reported to exhibit pharmacological properties such as antimicrobial,
antiviral, antitumor, anti-inflammatory and antioxidant activity
The fenugreek plant was included in normal diet generally because it has haematinic value

4. 2-Definition of endophtic fungi
Endophytic fungi are any fungi that grow all or part of their life
cycles symptomlessly in the intercellular spaces of living and
apparently healthy host plant tissues, while the inhabited host
tissues remain intact and functional (Petrini, 1991; Wilson, 1995;see Sieber, 2007; Hyde and
Soytong, 2008). However, the associations of endophytic fungi
with their host plants are varied and complex. These associations
are generally a cryptic phenomenon in nature. Endophytic fungi
often contribute to the normal health and development of their
hosts in exchange for a relatively privileged niche. Knowledge of
the extent of fungal endophytism is still relatively new, and we are
only starting to understand endophyte-plant interactions.

5. 3-Techniques for isolation and detection of fungal endophytes
1. Dilution-to-extinction cultivation of endophytic fungi, Recently extinction-to-
dilution has been suggested as an alternative method to fragment plating for
culturing fungal endophyte communities This preliminary study of
dilution-to-extinction was sufficiently successful that the authors advocated further testing
and optimization of extinction culturing in endophyte research. Perhaps the greatest
concern with use of the method is the effect of particle preparation on fungal hyphae. We
have noticed that some fungi, e.g., Neotyphodium endophytes in grass stems and
basidiomycetes in woody tissues are sensitive to the mechanical disruption and not readily
recovered in extinction cultures
2. Scraping method for culms or flowering stems,
Seed stems (culms) may be used for endophyte detection, where the endophyte is present
intercelularly the pith (parenchymatic tissue). Culms are cut into small segments 3-5 cm
long, then each segment is cut longitudinally in half, and central pith is scraped with a
scalpel and transferred to a slide over a drop of aqueous aniline blue dye. Then, observed
under a compound light microscope for the presence of typical hyphae running parallel to
the culm axis (Fig. 5A). However, this technique is limited to mature plants flowering
plants during a certain time of the year. This technique can also be used for tillers and leaf
sheaths (see Clark et al., 1983 for details of this technique).

6. 3.Scraping method for tillers and rhizomes
Individual tillers may be attached to a piece of the crown or base of the plant. The tiller is
cut longitudinally in half and the basal meristematic tissue is scrapped with a scalpel. When
using rhizomes, the rhizome is cut longitudinally in half and the pith is scrapped for
observation. The plant tissue is transferred to a glass slide on a drop aniline blue or rose
Bengal dye, then, observed under a compound light microscope for the presence of typical
hyphae (Fig. 5A). This technique is more appropriate for plant species with big tillers such
as tall fescue. Other techniques for direct visualization involve examination of seeds.
4.The seed squash method is used for observation of endophytes in seed material. Seeds are
pretreated with a 5% solution of sodium hydroxide (NaOH) for 8 hours at room
temperature to digest the seed coat and then rinsed several times with tap water. Individual
seeds are transferred to a glass slide and glumes are discarded using forceps. Seeds are
squashed under a coverslip after adding a drop of aqueous aniline blue or rose Bengal (for
stain particulars see Saha et al., 1984). Slides are examined under a compound light
microscope for the presence of typical non-branching mycelia surrounding the aleurone cell
layer (Fig. 5B). Pretreatment can be shortened to 30 min by repeating 2-3 times warming
up the sodium hydroxide solution until boiling point using a beaker partially covered in the
microwave oven. The seed squash technique is a useful and widely used technique for
fungal endophyte detection not only for fresh material but also for herbarium material and it
gives a reliable estimate of the presence of endophyte (Clark et al., 1983).