ISSN-0974-1550

ENVIS NEWSLETTER

MICROORGANISMS AND ENVIRONMENT MANGEMENT (Sponsored by Ministry of Environment, Forest & Climate Change, Government of India)

VOLUME 15 ISSUE 3 Jul. - Sep. 2017

ENVIS CENTRE Department of Zoology

University of Madras, Guindy Campus, Chennai - 600 025

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Cover page : Honey mushroom fungus Armillaria sp. The largest organism in the world, when

measured by area.

ISSN - 0974 - 1550

Volume 15 | Issue 3 | Jul. - Sep. 2017

EDITORS

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(ENVIS Coordinator)

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(Scientist - D)

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ENVIS Newsletter Vol.15/ Issue 3/ Jul. - Sep. 2017

1

CONTENTS Page

No. From the Coordinator’s Desk!

Dear Readers,

Greetings!

Search for new compounds in biotechnological and

biomedical sectors has increased. Even alternate sources for

the existing compounds are in demand as the current

production of some compounds is insufficient. Of the various

sources explored, the eukaryotic microorganisms fungi, offer

promising solutions. The fungi in the environment are utilized

mainly for bioremediation, bio fertilizers, food crops

development etc. The bioactive compounds of fungi are used

in treatment of infectious disease, cancer and antibiotic

development etc. The diversity of fungi in ecosystem is vast

and to understand their interaction with other microorganisms,

plants and animals is essential for innovative and effective

developments.

In this context, the present issue contains a research

article that explains about stimulation and production of taxol

from a fungus with other interesting topics such as alternative

antimicrobial compounds from wastewater, how microbes may

influence our behavior, use of fungi for assessing radioactivity

of environment and many more. Hope this issue would bring

an awareness on the importance of the fungi and their

associated studies.

Dr. C. Arulvasu

Kindly send your feedback @

www.envismadrasuniv.org/send_feedback.php

For further details, visit our website.

www.dzumenvis.nic.in: www.envismadrasuniv.org

SCIENTIFIC ARTICLE

Stimulation of taxol production by

Pestalotiopsis breviseta (sacc.) Steyaert using

biotic and abiotic elicitors Kathiravan*. G, Arulvasu. C, Meenashree. B and

Manickamoorthi. N

2

RESEARCH REPORTS

Alternative antimicrobial compounds could

come from wastewater

6

Fungi that evolved to eat wood offer new

biomass conversion tool

7

How microbes may influence our behavior 8

ONLINE REPORTS

Storming the cellular barricades to fight

fungi

9

Hunt is over for one of the top 50 most-

wanted fungi

9

NEWS

Fungi are key players of the deep biosphere 11

Fungi can be used as biomonitors for

assessing radioactivity in our environment

11

ABSTRACT OF RECENT

PUBLICATIONS

12

E - RESOURCES

EVENTS

SCIENTIFIC ARTICLE

Stimulation of taxol production by Pestalotiopsis breviseta (sacc.) Steyaert using

biotic and abiotic elicitors Kathiravan. G1*, Arulvasu. C2, Meenashree. B3 and Manickamoorthi. N3

1Department of Botany, Ramakrishna Mission Vivekanandha College, Chennai. 2 Department of Zoology, University of Madras, Chennai. 3Department of Biotechnology, Vels University,

VISTAS, Chennai

Keywords:

Fungal Taxol

Elicitors

Anticancer Drug

Biosynthesis

2

ABSTRACT

Coelomycetous fungi Pesalotiopsis breviseta (Sacc.) Steyaert was screened for the production of taxol an

anticancer drug. Taxol production was confirmed by Ultra Violet (UV) spectroscopy, Infra-Red (IR) spectroscopy,

High Performance Liquid Chromatography (HPLC) and Liquid Chromatography Mass Spectrum (LC - MS)

analysis. The compound produced by the fungus with and without elicitors was compared with authentic taxol,

identified and confirmed as taxol. Minimum of 150 µg/L taxol was produced with biotic fungal elicitor, while 160

µg/L was produced without any elicitors. Maximum of 250 µg/L taxol was produced with abiotic elicitor, copper

sulphate (CuSO4). The tested fungus P. breviseta, has a great potential for commercial exploitation in the future, for

better taxol production.

Introduction

Taxol is the commercial trade name used by Bristal-

Myers Squibb for the compound, paclitaxel. It has poor

water solubility and a complex chemical structure. Taxol

along with a chemically similar analogue, Taxotere

(Docetaxel) (Guenard et al., 1993) has unique anti-tumour

mechanism of action. The drugs are believed to block cell

cycle progression during mitosis by binding to and

stabilizing microtubules (Schiff et al., 1979, Nicolaou et al.,

1994). During cell division, taxol interferes with the

development of the microtubules needed for cell duplication

thus inhibiting the faster growing tumor cells. This is

different from other anti-cancer agents that work by

interfering with the DNA of tumor cells.

Clinical development of taxol progressed slowly

because of the extremely small amount of drug obtained

from the crude bark extract of the tree Pacific yew. Taxol

was approved for the treatment of ovarian cancer by the

Food and Drug Administration (FDA) in 1992 and

subsequently in the same year was used for treatment of

breast cancer. Since then clinical use of taxol had increased

and its use has been extended for treatment of lung cancer,

squamous cancers of the head and neck and various other

cancers (Eisenhaver and Vermorken, 1998). Naturally

Pacific yew grows slow but under intensive nursery culture,

can grow quite quickly. The tree lives 200-300 years with

some species 400 years or more and Taxol is found in small

quantities (0.001 to 0.01% of the dry bark weight) and the

content can vary in them. It is generally

considered to take 3 to 10 trees per patient. Based on the

year 1992 data from the US Forest Service, 36000 trees are

required to provide 327200 kg of bark (about 9 kg/tree)

from which only 24 kg of taxol can be extracted (about 0.66

g/tree). Hence there is an urgent need to search for the

alternative sources for this promising anticancer drug.

The plant cell culture of Taxus species appeared to be

promising to obtain taxol and related taxane compounds.

Ketchum and Gibson (1996) investigated this using free and

immobilized cells of Taxus cuspidata using perfusion

culture reactors and achieved continuous taxol production at

a rate of 0.3 mg/g DCW (dry cell weight) per day for 40

days.

The presence of various microorganisms from the bark

of the yew tree was found to be capable of producing taxol.

The fungi Taxomyces andreanae (Stierle et al., 1993)

isolated from the inner bark of a yew tree growing in North-

western Montana was able to produce taxol and other

taxanes in denovo fashion when grown in semi-synthetic

liquid media but the yield was very low (24-50 ng/L).

Pestalotiopsis microspora, a coelomycetous endophyte in

Taxus brevefolia was observed to produce taxol (Stroebel et

al., 1996) in considerable amount more than the Taxus sp.

There are atleast 12 distinct enzymatic reactions involved in

taxol biosynthesis (Croteau et al., 1995; Floss and Mocek,

1995). The genetic manipulation of fungi could be achieved

more easily than that of plants and it may be possible to

improve the production significantly with the help of

genetic engineering. Keeping this in mind there is an urgent *Corresponding author.

E-Mail address: kathir14journals@gmail.com

3

Thin layer chromatography (TLC):

Chromatography was done on 0.25 mm Merck precoated

silica gel plates. The plates were developed successively in each

of the following solvent systems as follows; A - Chloroform/

Methanol 7:1 v/v, B - Chloroform/ Acetonitrile 7:3 v/v, C - Ethyl

acetate/2-Propanol 95:5 v/v, D- Methylene chloride/ Tetra hydro

furan 6:2 v/v and E- Methylene chloride/ Methanol/

Dimethylformamide 90:9:1 v/v/v. Taxol was detected with 1%

w/v vanillin/sulphuric acid (H2SO4) reagent after gentle heating

(Cardellina, 1991). It appeared as a bluish spot fading to dark

grey after 24 hours.

Ultra Violet (UV) spectroscopy

The sample containing taxol was analysed

spectroscopically. After chromatography, the area of plate

containing putative taxol was carefully removed by scrapping

off the silica at the appropriate Rf and exhaustively eluted with

methanol. The UV spectral analysis of fungal sample was

superimposed on authentic taxol at 273 nm.

High Pressure Liquid Chromatography (HPLC)

Isolated samples were analysed by HPLC (Shimadzu 9A

model) using a reverse phase C18 column with a UV detector.

Sample (20 µl) was injected each time and detected at 232 nm.

The mobile phase was methanol/acetonitrile/water (25:35:40

v/v) at flow rate of 1.0 ml min-1. Taxol was quantified by

comparing the peak area of the samples with that of the standard

using the formula,

Taxol Content = Standard concentration × Total area of the sample

Total area of the standard

Infra-Red (IR) spectroscopy

The IR spectra of the compound were recorded on

Shimadzu FT IR 8000 series instrument. The purified taxol was

ground with spectra grade potassium bromide (KBr) (1:10)

pressed in to pellets under vacuum using spectra lab Pelletiser

and compared with authentic Taxol. The IR spectrum was

recorded in the region 4000 – 500 cm-1nm.

Liquid Chromatography - Mass Spectrometry

The sample from Pesalotiopsis breviseta was subjected to

electron spray mass spectroscopic analysis for the confirmation

of presence of taxol. The sample was dissolved in

methanol:water:acetic acid (50:50:1 v/v). It was injected with a

spray of 2 µL and spray voltage of 50 V by the loop injection

method.

need to search for the alternative sources for this

promising anticancer drug. The present study deals with

the stimulation of selected endophytic fungi Pesalotiopsis

breviseta for the production of taxol.

Materials and methods

Experimental designing

Biotic and abiotic elicitors were used for enhancement

of taxol production from the fungus. The taxol produced

without any elicitors by the fungus served as control. One

week after the inoculation of fungi in M1D broth media,

the abiotic elicitor copper sulphate (CuSO4) 50 mg/L was

added. Later after two weeks another dose of the abiotic

elicitor of same concentration was elicitated. For biotic,

fungal elicitors was used. Initially the fungi were

inoculated in M1D broth media without sucrose for a week

after which it was elicitated using 50 mg/L of fungal

carbohydrate.

Extraction of taxol

The extraction procedure was followed by the

method of Strobel et al. (1996). The fungus was grown

in 2 litre Erlenmeyer flasks containing 500 ml of M1D

medium as described in experimental setup and

supplemented with 1g soytone L-1 (Pinkerton and

Strobel, 1976). After incubation for 21 days at 26 ± 1°C

the culture filtrate was obtained by passing through

four-layered cheesecloth. To avoid fatty acid

contamination 0.25 g of sodium carbonate (NaCO3) was

added to the filtrate and extracted with equal volumes of

solvent dichloromethane twice. The organic phase was

collected and evaporated to dryness under reduced

pressure at 35ºC. The dry solid residue was re-dissolved

in methanol and loaded on a 1.5 X 30 cm column of

silica gel (Baker 40 micron). Elution of the column was

performed in a stepwise manner starting with 70 mL of

100% methylene chloride followed by mixtures of

organic solvents methylene chloride:ethyl acetate at

different proportions 20:1, 10:1, 6:1, 3:1 and 1:1 v/v.

Fractions were subjected to TLC and those having same

mobility as the authentic taxol were combined and

evaporated to dryness. The compound obtained was

analysed further for the presence of taxol in the fungal

samples.

4

Results and Discussion

In the present study the test fungus Pestalotiopsis

breviseta a Coelomycete, was tested for production and

enhancement of taxol by biotic and abiotic elicitors in a

semi-synthetic medium M1D, for its growth. The taxol

was extracted using dichloromethane and the solvent

was removed by evaporation under vacuum, the

resulting residue was re-dissolved in methanol and

treated as the sample containing taxol to further studies

for its confirmation.

Thin layer chromatography (TLC) was developed

on a 0.25 mm (10 x 20 cm) silica gel plate in different

solvent system with authentic taxol (Sigma, Cat. No. T-

7402) as standard. The sample showed identical Rf and

UV characteristics with the standard and reacted

positively with Vanillin/H2SO4 spray reagent, yielding a

blue spot which turned grey after 12 - 24 h. After

chromatography the taxol was eluted with methanol.

The UV spectral analysis of samples were

superimposed on that of authentic taxol with two

maxima at 273 nm and 235 nm which showed that the

purified sample might to be a taxol (Fig.1a-d).

amount of taxol produced by the fungus was quantified by

comparing the peak area of the samples (Fig. 2a-d) with that of

the taxol standard using the methods and formula described

earlier. The effect of biotic and abiotic elicitors on the fungus

Pestalotiopsis breviseta for taxol production was quantified.

The maximum taxol production was observed in the medium

containing chemical elicitor CuSO4 (250 μg/l) and minimal of

150 μg/L of taxol in medium containing fungal elicitor. The

cultures grown without any elicitors i.e. control produced 160

µg/L. Thus further convincing evidence for the identity of taxol

was obtained by High Pressure Liquid Chromatography.

Fig. 1: Ultra Violet (UV) Absorption of standard taxol

against taxol isolated from the fungus under different

conditions (control, biotic and abiotic elicitors)

The sample containing taxol was analysed by HPLC

using a reverse phase C18 column with a UV detector for the

quantification using the authentic taxol as standard. The

Fig. 2: High Pressure Liquid Chromatography (HPLC) analysis

of standard taxol against taxol isolated from the fungus under

different conditions (control, biotic and abiotic elicitors)

The presence of taxol was further confirmed by using

Infra-Red analysis of the compound isolated from the fungus.

The appearances of bands convincingly illustrated the identical

feature of the taxol isolated from the fungus without any

elicitor (Control) (Fig. 3b) and taxol isolated from the fungus

treated with abiotic elicitor (Fig. 3c) with the authentic taxol

used as standard (Fig. 3a). Infra-Red analysis showed that the

presence of alcoholic O - group in the sample is evident by its

OH stretching vibration at 3448 cm-1. The aliphatic CH -

stretch at 2931 cm-1, the C=O stretch positioned 1724 cm-1

and the amide C=O stretch is shifted to lower value at 1652 cm-

1. The intense peak

at 1247.16 cm-1 is due to COO stretch. The alkyl C-O stretch of

ester is observed at 1072 cm-1. The peak at 707 cm-1 is due to

aromatic C, H bond. In extracted sample though the intensity of

the bands are very much diminished in the fingerprinting region,

appearance of overtone 2362 cm-1 convincingly illustrates the

identical nature of the extracted sample with authentic taxol. The

taxol isolated from fungus treated with biotic elicitor was not

subjected to further analysis as its yield was very low

comparatively.

Fig. 3a&b: (a) IR spectrum of Authentic Taxol and (b) taxol

produced by Pestalotiopsis breviseta without elicitors (Control).

Fig. 3c: IR spectrum of Taxol extracted from Pestalotiopsis

breviseta using abiotic elicitor.

The fungal compound produced an identical LC mass

spectrum when compared with the standard. Characteristically,

authentic taxol yielded both an (M+H)+ peak at 855 and an

(M+Na)+ peak at 856 (Fig. 4a) and by comparison, fungal taxol

produced in the presence of abiotic elicitor also had these

characteristics peak (Fig. 4b). Based on these results it was

confirmed that the compound produced by the fungus

Pestalotiopsis breviseta was taxol.

Fig. 4a: Liquid Chromatography-Mass Spectrometry (LC-MS)

analysis of standard taxol

Fig. 4b: Liquid Chromatography-Mass Spectrometry (LC-MS)

analysis of taxol isolated from Pestalotiopsis breviseta treated

with abiotic elicitor.

The amount of taxol produced by the fungus could be

increased by improving culture techniques, addition of various

elicitors, application of genetic engineering and gene

expression studies. This would lead to improved production of

taxol and overrule its production from plant source.

References

Cardellina, J. H. (1991). HPLC separation of taxol and

cephalomannine. J. Liq. Chromatogr. 14 (4): 659 - 665.

Croteau R, Hefner J, Hezari M , Lewis NG: Flores, H. E. and

Gustine. D. I. (Eds.) (1995). American society of plant

physiology. Phytochemicals and Health.

Eisenhauer, E. A. and Vermorken, J. B. (1998). The taxoids.

Drugs. 55 (1): 5 - 30.

Floss, H. G. and Mocek, U. (1995). Biosynthesis of taxol. In

Taxol: science and applications. CRC Press, Boca Raton,

Florida. pp. 191-208.

Guenard, D., Gueritte-Voegelein, F. and Potier, P. (1993).

Taxol and taxotere: discovery, chemistry, and structure-

activity relationships. Acc. Chem. Res. 26 (4): 160 - 167.

Ketchum, R. E. B. and Gibson, D. M. (1996). Paclitaxel

production in suspension cell cultures of Taxus. Plant Cell,

Tissue Organ Cult. 46 (1): 9 - 16. 5

Nicolaou, K. C., Yang, Z., Liu, J. J., Ueno, H., Nantermet, P. G.,

Guy, R. K. and Sorensen, E. J. (1994). Total synthesis of

taxol. Nature. 367 (6464): 630 - 634.

Pinkerton, R. and, Strobel, G (1976).: Serinol as an activator of

toxin production in attenuated cultures of H. sacchari. Proc.

Natl. Acad. Sci. USA. 1976; 73: 4007 - 4011.

Schiff, P. B., Fant, J. and Horwitz, S. B. (1979). Promotion of

microtubule assembly in vitro by taxol. Nature. 277 (5698):

665 - 667.

Stierle, A., Strobel, G. and Stierle, D. (1993). Taxol and taxane

production by Taxomyces andreanae, an endophytic fungus of

Pacific yew. SCIENCE-NEW YORK THEN

WASHINGTON, 260: 214 - 214.

Strobel, G., Yang, X., Sears, J., Kramer, R., Sidhu, R. S. and Hess,

W. M. (1996). Taxol from Pestalotiopsis microspora, an

endophytic fungus of Taxus wallachiana. Microbiol. 142 (2):

435 - 440.

RESEARCH REPORTS

Municipal wastewater may become a key ally in the fight

against antibiotic-resistant disease-causing bacteria and fungi, a

new study has found.

According to Dr Thando Ndlovu, a postdoctoral researcher in

the Department of Microbiology at Stellenbosch University,

certain bacteria present in municipal wastewater produce

antimicrobial compounds or biosurfactants that can help to

prevent the growth of antibiotic-resistant microorganisms which

cause serious infections in humans. Ndlovu had recently obtained

his doctorate in Microbiology at Stellenbosch University under the

supervision of Prof. Wesaal Khan from the same department.

He said that the rapid increase in the emergence of antibiotic-

resistant bacteria was a major reason behind his search for new

antimicrobial compounds. As part of his research, Ndlovu

collected wastewater samples and also carried out molecular and

microbiological tests in a laboratory on various biosurfactants-

producing bacteria found in these samples. He isolated two

bacterial strains whose biosurfactants proved effective against

antibiotic-resistant disease-causing bacteria. Biosurfactants are

compounds produced naturally by bacteria, fungi or yeasts and

they have been commercially utilized in shampoos, shower gels,

and household cleaning products. They are also used in food,

agriculture, cosmetic and medical industries as well as in

environmental bioremediation to prevent the spread of

spoilage and disease-causing bacteria.

The biosurfactants produced by two bacteria in his study

prevented the growth of major disease-causing bacteria such

as methicillin-resistant Staphylococcus aureus and

gentamicin-resistant E. coli which can lead to life-threatening

infections in humans. This finding is promising as worldwide

reports on the number of deaths caused by antimicrobial

resistant microorganisms that are becoming increasingly

difficult to treat with currently available drugs.

The discovery of novel antimicrobial compounds is a

priority and biosurfactant compounds could be used to

develop new antibiotics for treatment of various infections

caused by antibiotic resistant bacteria and eventually replace

ineffective antibiotics in future. Ndlovu showed in his study

that municipal wastewater is ideal for the isolation of diverse

biosurfactant-producing bacteria that could be utilized in the

production of such compounds for commercial use.

While numerous studies have reported on the isolation of

biosurfactant-producing bacteria from contaminated soil and

terrestrial environments, the current study indicated that

municipal wastewater could be exploited for the isolation of

diverse biosurfactant-producing bacterial strains.

Biosurfactant-producing bacteria thrive in polluted

environments such as contaminated soil or water. These

bacteria also have the ability to outcompete other bacteria in

the same environment because the biosurfactant compounds

help them to absorb nutrients and to protect them from toxic

materials.

Fig. South African microbiologist, Dr. Thando Ndlovu in his

laboratory in the Department of Microbiology, Stellenbosch

University.

Image credit: Stefan Els

6

Alternative antimicrobial compounds

could come from wastewater

Ndlovu adds that biosurfactant compounds can be used to

reduce the use of synthetic antimicrobial agents for various

purposes such as cleaning and coating agents to prevent the

build-up of disease-causing and spoilage bacteria. As far as

future research is concerned, Ndlovu said that he is now focusing

on the application of biosurfactant compounds with antimicrobial

properties.

Source: www.sciencedaily.com.

Twenty years ago, a microbiologist and his colleagues

discovered a unique system that some microorganisms use to

digest and recycle wood. Three orders of brown rot fungi have

now been identified that can break down biomass, but details of

the mechanism were not known. Using several complementary

research tools, Goodell and colleagues reported new details of

this unexpected mechanism at work, one that surprisingly does

not involve enzymes, the usual accelerators of chemical

reactions. Basidiomycota brown rot fungi use a non-enzymatic,

chelator-mediated biocatalysis method that is very different than

that used by any other microorganism studied. Chelators are

organic compounds that bind metal ions and in this case they also

generate hydroxyl radicals to break down wood and produce

simple building-block chemicals.

Described by collaborators at Oak Ridge National

Laboratory as a paradigm shift in understanding fungal

biocatalysis for biomass conversion the findings appear in the

journal of Biotechnology for Biofuels. Goodell said their research

on fungal bioconversion systems looks at a novel mechanism that

has potential use in bio-refineries to deconstruct woody biomass

for conversion into platform chemicals for biopolymers or energy

products.

Brown rot fungi appear in both the northern and southern

hemispheres and are some of the most common decay fungi in

North America. Because they evolved relatively recently, there

are fewer brown rot species compared to older white rot species.

Goodell said that because of their rich efficiency in degrading

wood particularly in degrading softwoods. Brown rot fungi have

now dominated by recycling approximately 80 percent of the

softwood biomass carbon in the world, found mostly in the great

forests of the northern hemisphere.

Goodell points out that most microorganisms use enzymes

to break down compounds, but enzymes are huge molecules

and physiologically expensive to produce because they contain

so much nitrogen. He noted that scientists used to think that

these fungi would make holes in the cell wall that would let in

the big enzymes a sort of pretreatment model. But now they

have explained how it works. The fungi in their study used a

non-enzymatic, catalytic chelator-mediated Fenton system, a

very simple process that makes use of hydrogen peroxide

which is also generated by the fungal system and iron found in

the environment. He adds that he and his colleagues believe the

brown rot fungi's efficiency comes from their use of the

chelator-mediated Fenton system rather than the use of

enzymes exclusively, as white rot fungi do.

Goodell noted that this group of brown rot fungi generates

hydroxyl radicals at a distance that is away from the fungus so

the radicals won't damage themselves while breaking down

wood. Hydroxyl radicals are very damaging to cells, the most

potent oxidizing agents known in biological systems.

For this work, Goodell and his colleagues including his

collaborator Jody Jellison, now director of the Center for

Agriculture, Food and the Environment at UMass Amherst,

used a suite of investigative methods including Small Angle

Neutron Scattering (SANS), Sum Frequency Generation (SFG)

Spectroscopy, Fourier Transform Infra-Red (FTIR) analysis,

X-Ray Diffraction (XRD), Atomic Force Microscopy (AFM)

and Transmission Electron Microscopy (TEM) to fully

describe the process.

Goodell said that these fungi do produce a limited number

of enzymes but they come into play after the non-enzymatic

action conversion by the fungi using chelators. The chelators

are secondary metabolites, whose function is not easily

followed using omics techniques such as genomics. Using

many advanced techniques they found some very small, low-

molecular-weight compounds working their way into the cell

wall.

Goodell and Jellison relate a process that begins with the

fungi in the lumen - the hollow space found inside plant cells.

Using their hyphae, thread-like growth filaments, the fungi

then mount a biochemical attack on the wood cell components.

They explained that this group of fungi evolved a way to

break down the wood substrate by first diffusing chelators into

the cell wall. The fungus makes the chelator and produces 7

Fungi that evolved to eat wood offer

new biomass conversion tool

hydrogen peroxide from oxygen and together they start to digest

the cell wall into the sugar found in the basic building block of

wood, glucose, which the fungus uses as food. This is how these

fungi are eating the wood.

Source: www.sciencedaily.com.

Stress, anxiety and depression are emotions we all feel at

some point in our lives and some people feel them to a greater

degree than others. Part of the human experience, right?

In a recent study, John Cryan, a researcher at the APC

Microbiome Institute at University College Cork in Ireland and

his colleagues reported a link between the microbiome and fear.

By examining mice with and without gut bacteria, they

discovered that the germ-free mice had blunted fear responses

(Mol. Psychiatr. doi:10.1038/mp.2017.100, 2017). Their findings

may pave the way for the development of novel treatments for

anxiety-related illnesses, including posttraumatic stress disorder.

“It may seem odd that my research focuses on the gut if I‟m

interested in the brain. But when we think of how we express

emotion in language, through sayings like „butterflies in your

tummy‟ and „gut feeling,‟ it isn‟t surprising that they‟re

connected” says John.

Researchers at Kyushu University in Japan were the first to

show in 2004, that bacteria in the gut can influence stress

responses, prompting many subsequent investigations. Yet

despite mounting research scientists remained uncertain about

exactly how the gut microbiome affects the brain. While some

bacteria influence the brain through the vagus nerve other strains

seem to use different pathways. It is known that the population of

the gut microbiome begins in early life, and recent research

suggests that disruptions to its normal development may

influence future physical and mental health (Nat. Commun.

6:7735, 2015).

Researchers are finding that this gut-brain connection could

have clinical implications as influencing the gut microbiome

through diet may serve to ameliorate some psychiatric disorders.

Together with University College Cork colleague Ted Dinan,

Cryan coined the term “psychobiotics” in 2013 to describe live

organisms that when ingested produce health benefits in patients

with psychiatric illness. These include foods containing

probiotics and live strains of gut-friendly bacteria.

While there are many rodent studies linking probiotics and

mental health, UCLA biologist Emeran Mayer and his

colleagues were the first to test them in humans, using

functional magnetic resonance imaging (fMRI) scans to assess

the results. After administering probiotic yogurt to a group of

healthy women twice a day for four weeks, the researchers

found that the women had a reduced brain response to negative

images (Gastroenterology, 144:1394-401, 2013). Having

conducted this study on healthy participants, Mayer is reluctant

to conclude that probiotics can cure mental illnesses such as

anxiety. He said it to be a complex emotion, not just a reflex

behavior like in the mouse. However, Mayer told he is very

supportive of the potential of prebiotics, fiber-rich foods that

promote the growth of beneficial bacteria in the gut.

Researchers at Deakin University in Australia recently

trialed a Mediterranean-style diet, which is predominately

plant-based and fiber-rich in a group of adults with major

depression. They found that one-third of the participants

reported a significant improvement in symptoms after 12

weeks on the diet (BMC Medicine, 15:23, 2017).

Just as activity in the gut seems to affect the brain, mental

stress can lead to intestinal problems. Scientists have

demonstrated this in research on irritable bowel syndrome. For

example, a study by Mayer and colleagues linked early-life

emotional trauma to an increased risk of developing the bowel

disorder (Clin. Gastroenterol. Hepatol. 10:385-90, 2012).

As data on the brain-gut axis accumulates, many scientists

are taking notice. Trinity College Dublin researcher Shane

O‟Mara said that there is a great potential in this area but

cautioned that it is too early to say whether targeting the

microbiome will play a role in psychiatric treatment.

University of Manitoba gastroenterologist Charles Bernstein

also feels the research is promising but believes that we are far

from manipulating the microbiome to treat mental health

disorders. Those spearheading this research are equally aware

of the need for more studies, particularly in human subjects,

but they are hopeful that change lies ahead.

“I‟m almost certain that in several years, diet will be

considered one branch of therapy for many mental illnesses,

alongside medication and psychiatric treatments,” says Mayer.

“People with severe mental illness will still need something

very strong, but this is a useful adjunctive,” agrees Cryan. “I

think when we go to our GP in future, we will not only have 8

How microbes may influence our

behavior

blood tests, we will have the microbiome tested.”

“Within five years, I hope to see more clinical trials that

demonstrate the efficacy of prebiotics and probiotics on mental

health disorders,” says University of Chicago microbial ecologist

Jack Gilbert. “There needs to be a revolution in how we deal with

mental illness in our society.”

Source: www.the-scientist.com

ONLINE REPORTS

Yale scientists have developed a new class of small

molecules that attack fungal cell wall of pathogenic fungi and

signaling antibodies to join the process. The discovery offers a

potential new therapeutic approach to treat fungal illnesses that

affect thousands of people each year, including patients whose

immune systems are compromised by organ transplants, cancer

treatment and HIV infections.

The new compounds are called antibody-recruiting

molecules targeting fungi (ARM-Fs). These small molecules

have two main features: a target-binding terminus that latches

onto the fungal cell wall and an antibody-binding terminus that

recognizes and recruits antibodies already present in the human

bloodstream. Using the human immune system as the effector

arm this strategy was found to be incredibly versatile said Yale

chemistry and pharmacology professor David Spiegel and senior

author of the study describing the discovery in the German

science journal Angewandte Chemie. This is the first time they

have shown this strategy to work in treating a fungal disease.

Over the past decade, Spiegel's lab has explored small-

molecule approaches in treating a range of diseases, including

cancer and HIV. Spiegel said that not only are such molecules

effective against drug-resistant strains of diseases but also may

be used in combination with existing treatments.

Fig. The design showing antifungal antibody-recruiting small

molecules (ARM-F) targeting chitin, a fibrous substance in the

cell walls of fungi.

Image credit: Yale University

Source: www.sciencedaily.com

In a step toward bridging the gap between fungal

taxonomy and molecular ecology, scientists have characterized

a sample of mystery fungus collected in North Carolina and

found its home in the fungal tree of life.

"Working estimates tell us that there should be more than

5 million species of fungi," said Cheryl Kuske, a Los Alamos

scientist on the project. They have identified and fully

described 100,000 of them, though new DNA sequencing

capabilities showed that many specimens in their research

collections are uncharacterized. Solving this particular mystery

would reveal the potential value of using environmental

sequencing to guide taxonomic and ecological discovery.

The fungal sample was interesting as it partly represented

a major component of the observed fungal population in a pine

forest and it responded positively to elevated CO2 and nitrogen

amendment treatments that mimic future environmental

conditions yet the sample's exact placement in the taxonomic

order was unknown. The sample was eventually given a new

name, Bifiguratus adelaidae as reported recently in the journal

Mycologia.

Why was this hard? In times when a simple cheek swab

mailed off with a check can produce a human DNA report

listing thousands of ancestors, a sample identification would

seem a simple task. But as an editorial in the journal pointed

out, this sample represented one of many dark matter fungi that

populate unknown regions of the fungal tree of life. We can

detect their DNA in environmental samples, but their culture

has been elusive.

As noted in the journal's editorial, "This placement is

particularly exciting because of the increased understanding of

the mycorrhizal role (ability to form symbiotic, nutrient-

transfer relationships) for this part of the phylogeny and the

fact that very little is known about the species diversity and

distribution in this part of the tree. Bifiguratus adelaidae may

have a symbiotic function in roots, having been detected in 9

Storming the cellular barricades

to fight fungi

Hunt is over for one of the 'top 50

most-wanted fungi'

orchid and chestnut roots, but it is also well-documented in soils

from north temperate zones."

Interestingly, the collected sample had been cultured

successfully in the laboratory only when it was allowed to grow

in the company of a species of bacteria, Methylobacterium,

which antibiotics were unable to kill. Normally the fungal sample

would have been cleared of such contaminants, but this one

resisted their attempts. The authors posit that the symbiotic or

commensal microbes presence may be part of the culturing

success. Understanding complex microbial and fungal

communities is progressing, Kuske noted, by allowing organisms

to grow as defined mixtures.

The Bifiguratus adelaidae was named in honor of Adelaida

Chaverri Polini. Dr. Chaverri was a world-recognized tropical

biologist and role model for women in Latin America in science,

technology, engineering, and mathematics. Her major

contributions were in the study and conservation of tropical

montane forests and treeless alpine grasslands (páramos). As a

biologist, she recognized the importance of mycorrhizal fungi

and contributed to the description of a fungus in the

Acaulosporaceae, a fungus in the same phylum as the species

described in this paper. She supported the creation of Costa

Rica's National Park Service, providing the basis for extensive

conservation of biological diversity. In 2013, she was included in

the Galería de la Mujer, one of the highest honors given to Costa

Rican women for their dedication to defending and improving

women human rights.

Fig. Researchers at Los Alamos National Laboratory and several

other institutions have characterized a sample of mystery fungus

and found its home in the fungal tree of life. The fungus's exact

placement among the taxonomic order was unknown until now.

Los Alamos National Laboratory conducts a wide range of

biological research efforts as part of its national security science

mission, with such research as phylogenetic analysis helping to

determine safe pathogens and disease transmission pathways.

Image credit: Los Alamos National Laboratory

Source: www.sciencedaily.com

10

Peroxisomes identified as 'fighters' in

the battle against bacterial infections

A new addition to the fight against bacteria comes in

the unlikely form of an organelle that previously had no link

to the immune response. University of Alberta researchers

have found that peroxisomes are required for the cells in the

innate immune response against bacteria and fungi. The

discovery was first made in fruit flies. Research Associate

Francesca Di Cara, together with Richard Rachubinski,

Professor and Chair of the Department of Cell Biology and

Andrew Simmonds, Cell Biology Associate Professor,

partnered to create fruit flies that could be used specifically

for studying peroxisomal disorders, which are rare genetic

diseases affecting humans.

Di Cara found that peroxisomes are necessary for

proper functioning of the innate immune system, the body's

first line of defense against microorganisms. The innate

immune system is an ancient system of immunity that

identifies, captures and processes a pathogen and then

presents it to the acquired immune system. The peroxisomes

also communicate to other organs that there is an infection.

The team discovered that when the organelle's basic

function is altered, this communication is lost and the

organism does not fight the bacteria.

"Understanding how the body fights infection has an

impact on human health," says Di Cara. "We have to

understand who the fighters in the organism are before we

can identify what's failing in the battle against bacterial

infections.“

Peroxisomes are chemical factories that process

complex fat molecules into simple forms and modify

reactive oxygen molecules, which together act to signal cells

and tissues to respond appropriately for the changes in their

environment. Along with their collaborator Nancy

Braverman from McGill University, the researchers used a

mouse model to confirm that what they observed in the flies

also occurred in a mammalian system.

"To find organelles like peroxisomes that had no link

whatsoever to fighting bacterial infections was a critical

discovery, it will help expand the roles of what this

important organelle does in innate immunity against

bacterial and fungi and its involvement in viral signaling and

the lethal peroxisome genetic diseases," says Rachubinski.

As the threat of bacterial infections continues to grow, this

discovery can help move our understanding of immunity

forward.

Source: www.sciencedaily.com

research. This new finding proposes that they may be key

players in this globally vast realm. Studies of subterranean life-

forms have implications for early life on our planet and for life

on other planets, where hostile conditions may have inhibited

colonization of the surface.

Source: www.sciencedaily.com

Radioactive contamination is the unwanted presence of

radioactive substances in the environment. Our environment is

contaminated by naturally-occurring and anthropogenic

radionuclides, unstable isotopes of an element that releases

radiation and as it decomposes it becomes more stable which

are present in the air, soil, rain, etc. These radionuclides can be

transferred throughout the food chain until reaching humans

and this could lead for a potential health risk.

Until now, to study the presence of radionuclides in

different products for human consumption and their subsequent

transfer, research has been based fundamentally on foods such

as meats, fish or milk, without considering a foodstuff like

fungi which are well known for accumulating concentrations of

some radionuclides in their fruiting bodies. As a result, the

Environmental Radioactivity Laboratory of the University of

Extremadura (LARUEX) had carried out a study to quantify

radioactive presence in this foodstuff. The author of the study

Javier Guillén explained that the quantification was made using

transfer coefficients that compare the radioactive content in the

receptor compartment of the radioactive contamination (that is

to say in the fungi) to that existing in the transmitter

compartment (which in this case would be the soil).

To conduct this research the authors considered the base

level of radionuclides established in ecosystems with low

radioactive content and then used the software called the

ERICA Tool which allows one to enter the transfer coefficient

from the soil to the organism thus calculating the dose of

radionuclides a non-human organism receives.

From the study, it may be concluded that the estimated

dose rates for fungi are similar to those determined for other

animals (animals and plants) and therefore this species can be

used when assessing for the presence or absence of radioactive

contamination in the soil. As a result, the researcher asserted

that even though it is not strictly necessary to include

11

NEWS

In addition to the life on the surface of the Earth and in

its oceans, ecosystems have evolved deep under the earth

coined by the term “deep biosphere” which stretches several

kilometers down into the bedrock. The conditions are harsh

without oxygen and other favorable conditions present in the

earth surface. The knowledge about ancient life in this deep

environment is extremely scarce and most studies so far have

focused on the prokaryotes. A new study by an international

team of researchers led by Dr. Henrik Drake of the Linnaeus

University and Dr. Magnus Ivarsson of the Swedish Museum

of Natural History sheds light on eukaryotes in this deep

setting. They present the first in situ finding of fungi at great

depth in the bedrock. This ancient life is found at 740 m

below the ground surface. It represents a new piece in the

deep biosphere puzzle.

Henrik Drake, lead author of the study discovered the

fungi in a cavity hidden within a vein in a drill core he was

examining. They were beautiful like mineral crystals and

abundant in mycelium of fungal hyphae. To him it was like

observing a small community frozen in time.

Magnus Ivarsson another author explained more about

the fungi. Their detailed synchrotron-based investigations

clearly proved that it is fungi adapted to anaerobic conditions.

The fungi are partly mineralized and partly organically

preserved, which a rare find is revealing how organisms in

that environment are fossilized and preserved.

High spatial resolution isotope analysis within the

minerals that occur along with the fungi revealed that a

variety of microbial processes had occurred in the caveat,

including methane consumption and sulfate reduction. The

fungi could not be dated precisely but there are proxies

pointing to an age of tens of millions of years.

The study confirms a previously hypothesized

consortium between fungi and sulfate reducing bacteria, a

coupling that has yet been unsupported by direct evidence in

nature. As fungi provide hydrogen gas that fuel prokaryotes,

the findings suggest a re-evaluation of the energy cycling

within the energy-poor deep continental biosphere.

Eukaryotes have been neglected in the deep biosphere

Fungi are key players of the deep

biosphere

Fungi can be used as biomonitors

for assessing radioactivity in our

environment

fungi amongst the existing instruments and frameworks of

assessment, they can be used in ecosystems which may require

them based on criteria such as biodiversity.

Moreover, in the cases of the fungi analyzed, which are

concentrated in the Mediterranean area it should also highlight

the fact that they do not contain a high dose of radionuclides,

meaning there is no environmental contamination and they are

therefore perfectly suitable for consumption by humans.

Fig. Fungi can be used when assessing the presence or absence

of radioactive contamination in the soil.

Image Credit: Uex

Source: www.sciencedaily.com

01Applied Soil Ecology, 2017

Biochar chemistry defined by 13C-CPMAS NMR

explains opposite effects on soilborne microbes and crop

plants.

GiulianoBonanomi, Francesca Ippolito, Gaspare Cesarano,

Francesco Vinale, Nadia Lombardi, Antonio Crasto, Sheridan

L.Woob, Felice Scala.

Department of Agricultural Sciences, University of Naples

Federico II, Via Università 100, 80055, Portici, NA, Italy.

Numerous recent studies have demonstrated that biochar

may significantly reduce the incidence of plant diseases caused

by airborne and soilborne pathogens, although contrasting

results have also been reported. In this work, we investigated

how biochar affects crop plant and soilborne microbe growth.

Aims of this study were: i) to analyze the chemical changes

occurring in four organic feedstocks (e.g. wood chips, organic

urban waste, Zea mays residues, and Medicago sativa hay)

when pyrolyzed at 300 °C and 550 °C by using 13C NMR

spectroscopy and SEM (Scanning Electron Microscopy); ii) to

assess how biochar affects growth of five bacteria, nine fungi,

and three crop plants; and iii) clarify the relationships

between biochar chemistry and its effect on target species.

As pyrolyzation temperature increased, organic matter

chemistry of all products changed significantly, with a

progressive loss of O-alkyl C, di-O-alkyl C, and methoxyl

and N-alkyl C, coupled with an enrichment in aromatic C

types. Untreated urban waste and Medicago hay severely

inhibited Lepidium, Lactuca and Solanum root growth,

whereas no inhibitory effects were found for the other

feedstocks. However, these phytotoxic effects largely

decreased after pyrolyzation. In contrast to the crop plants,

fungi and bacteria thrive on most of the unprocessed organic

materials but showed reduced growth and development or

complete growth inhibition on biochars obtained at 300 °C

and 550 °C. Soilborne microbes demonstrated remarkably

similar correlation patterns between their growth to the

organic feedstock and biochar chemical components. This

work demonstrates that defining organic matter quality by

13C NMR extends our understanding of the impact of biochar

on crop plants and key components of the soil food-web.

Keywords: Beneficial microbes; C/N and H/C ratios;

13C CPMAS NMR; Disease suppression; Organic

amendment; Phytotoxicity; Rhizoctonia solani; Soilborne

pathogens.

12

Abstract of Recent Publications

NATIONAL

Cental Institute of Freshwater Aquaculture (CIFA)

http://cifa.nic.in

Patent facilitating centre for microorganisms

http://www.pfc.org.in/ach/micro.htm

National Centre for Biological Sciences

https://www.ncbs.res.in

National Collection of Industrial Microorganisms

http://www.ncl-india.org/ncim

INTERNATIONAL

American Society for Microbiology

https://www.asm.org

Center for Microbial Ecology

http://www.cme.msu.edu

International Society for Molecular Plant-Microbe Interactions

https://www.ismpmi.org/Pages/default.aspx

Japan Collection of Microorganisms (JCM)

http://jcm.brc.riken.jp/en/

E - Resources

EVENTS

Conferences / Seminars / Meetings 2017-18

4th International Conference on Microbial Diversity 2017. October 24 - 26, 2017. Venue: Bari, ITALY. Website:

https://www.md2017.it/about/

International Society for Subsurface Microbiology (ISSM) 2017. November 06 - 10, 2017. Venue: Rotorua, New

Zealand. Website: http://www.subsurfacemicrobiology.com

SMi's Pharmaceutical Microbiology East Coast Conference. November 13 - 14, 2017. Venue: Iselin, NJ, USA.

Website: https://www.smi-online.co.uk/pharmaceuticals/northamerica/conference/Pharmaceutical-Microbiology-East-

Coast

International Conference on Fungal & Infectious Diseases. December 04 - 05, 2017. Venue: Dubai, UAE.

Website: https://fungalinfections.conferenceseries.com

Pharmaceutical Microbiology UK. January 22 - 23, 2018. Venue: London, UK. Website: https://www.smi-

online.co.uk/pharmaceuticals/uk/pharmaceutical-microbiology

Pause to read the traffic sign: Regulation of DNA transcription in bacteria

Source: www.Phys.org

One of the central tenets of biology is that information flows from DNA to RNA in order to encode proteins,

which function in the cell. Arguably just as critical as the genetic code is the timing of this information flow. By

producing the right RNA and right proteins at the right time, a cell can effectively strategize its survival and

success. One such regulatory element, the riboswitch, has excited interest as a potential target for antibiotics. After

over 10 years of research, Scientists of Goethe University together with colleagues from other universities have

put together the puzzle pieces of a riboswitch-based regulatory process in the bacterium Bacillus subtilis,

presenting the most extensive model of the timing of riboswitch action to date.

A riboswitch is a short piece of RNA that can fold into different

structures, depending on whether or not a small messenger molecule is

binds to it. In transcriptional riboswitches, these different structures

signal the nearby RNA polymerase to continue producing RNA or to

stop. In their recent publication in ELife, the Schwalbe group and their

collaborators released molecular structures of the xpt-pbuX riboswitch

in the off-position after synthesis and in the on-position upon binding by

the small messenger molecule guanine. They also demonstrated that this

switch to the on-position takes a certain amount of time. This sets a

certain requirement on this regulatory process.

World Ozone Day Celebrations – Workshop on “Caring for all

under the sun” held on September 16, 2017.