Experiential Learning Programmes

(2022-23)
Production technology for yeast (PTH 403)

Department of Plant pathology (Yeast),

School of Agriculture, Lovely Professional
University, Phagwara – 144111

1
 Preface
Students must evaluate their qualification skill and knowledge more prudently
because skills and capabilities are also important with ranks or grades.
Benjamin Franklin quotes, “tell me and I forget, teach me and I remember,
involve me and I learn”, this advocate that ‘Learning by doing’ and ‘earning
by learning’ is one of the most important pillars of future career development.
Under the new course curriculum of ICAR, New Delhi, Student READY (Rural
Entrepreneurship Awareness Development Yojana) program which is based on
Experiential Learning Programme (ELP) in agricultural education system has
been conceptualized for building skills in project development and execution,
decision-making, team coordination, with end-to-end approach to problem
solving, accounting, quality control and marketing. The biggest benefit of
hands-on training is the opportunity for repeated practice. Project work
component in ELP provides several opportunities to students to learn many
aspects that cannot be taught in a class room or laboratory. In order to provide
such opportunities to the graduates of agricultural science, students project one
of the important components of the Student READY. In this direction, we are
making every efforts by adhering to the concept and aims of ELP units being
implemented in School of Agriculture to impart training on all aspects and to
prepare our students to become entrepreneurs. Experiential learning modules
namely “Production technology for yeast” is commissioned efficiently at
Department of Horticulture, School of Agriculture, Lovely Professional
University. This compilation "An Overview on Experiential Learning
Programmes”, provides an elaborate view of this module.

Dr. Deewakar Baral

Assistant Professor and ELP Coordinator

Department of Plant Pathology,

School of Agriculture,

Lovely Professional University, Phagwara (Punjab) – 144111

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 Index

Sr. no. Description Page no.

1. Introduction 1-6
2. Commercial aspect in food 7-8
industry
3. Potential of the sector 9-10
4. Activities 11
5. Activities chart 12-
6. Photograps
7. Outcomes
8. List of the students
9. conclusions

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Experiential learning Programme - A Novel Educational Tool
The Concept
A novel method of education based on Experiential Learning Programme (ELP)
has been introduced in the agricultural education system through Indian Council
of Agricultural Research. Conceptualized for building skills in project
development and execution, decision-making, individual and team coordination,
approach to problem solving, accounting, quality control, marketing and
resolving conflicts, etc. the programme is designed with end-to-end approach. It
is an essential prerequisite for the award of B.Sc. (Hons) Agriculture degree has
been reorienting graduates of Agriculture for ensuring and assuring
employability and entrepreneur mindset. The word “experiential‟ essentially
means that learning and development are achieved through personally
determined experience and involvement, rather than on received teaching or
training, typically in group, by observation, study of theory or hypothesis, bring
in innovation or some other transfer of skills or knowledge. Experiential
learning is a business curriculum-related endeavor which is interactive.
Experiential Learning is for building (or reinforcing) skills in project
development and execution, decision making, individual and team coordination,
approach to problem solving, accounting, marketing and resolving conflicts, etc.
The programme has end to end approach. Carefully calibrated activities move
participants to explore and discover their own potential. Both activities and
facilitation play a critical role in enhancing team performance.

ELP Objectives

Experiential Learning provides the students an excellent opportunity to develop

analytical and entrepreneurial skills, and knowledge through meaningful hands-
on experience, confidence in their ability to design and execute project work.
The main objectives of ELP are:

• To promote professional skills and knowledge through meaningful hands-on

experience.
• To build confidence and ability to work in project mode.
• To acquire enterprise management capabilities.
• Learning the art and science of production and managerial skills under
protected environment.
• Thus, the basic objective behind this programme is to develop skilled man
power and entrepreneurship mindset.

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 Expected Benefits
Experiential Learning helps the student to develop competence, capability, capacity
building, acquiring skills, expertise, and confidence to start their own enterprise and
turn job creators instead of job seekers. This is step forward for earning while learning
concept.

Experiential learning (EL) is a philosophy and methodologies in which educators

purposefully engage with learners in direct experience and focused reflection in order
to increase knowledge, develop skills, entrepreneurship, and inculcate values. The
word „experiential‟ essentially means that learning and development are achieved
through personal involvement and experience, typically in group, by observation,
listening, study of theory or hypothesis, rather than on received teaching or training.
EL is a business curriculum related endeavor which is interactive.

Experiential learning is a powerful way to address individual growth and potential,

which is commonly a much-neglected approach to teaching and developing people of
all ages. Experiential learning implies growth of a person from the inside, whereas
conventional teaching and training is the transfer of capability into a person from the
outside. The modern Education system based on experiential learning is determined
and controlled by the individual for the purpose of achieving personal development
and growth, whereas traditional system is based on conventional training and teaching
is designed and delivered by an organization for the purpose of developing the
capabilities (usually knowledge and/or skills) of a group of people to achieve a known
measurable standard or qualification.

Looking to the practical applicability in a broader sense, the need of Experiential

Learning programme has been strongly realized in the Agricultural education system.
As ELP is learner centered, individually directed with flexible outcomes as compared
to conventional teaching programme which is training-centered, theory focused with
prescribed fixed format and syllabus. ELP based programme eradicates the problems
associated with lack of concentration and slow understanding process among students
while implies on long term interest generation along with engaging students in
learning while actual participation in the task. Thus, Experiential Learning is a major
step forward for High Quality Professional Competence, Practical Work Experience in
Real Life Situation to Graduates, Facilitates producing Job Providers rather than Job
Seekers and Entrepreneurial Orientation. ELP holds great promise and potential for
managerial, technical and entrepreneurial skill development in the field of agriculture.

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As per ICAR, IVth Deans’ recommendation the ELP programme is
offered to the students during the VII semester for a total duration of
24 weeks with a weightage of 0+10 credit hours.
Component and Major Activities Credit Semester Departments
PTH403 – Experiential Learning 0+10 7th Department of plant
(Yeast) pathology, School of
Agriculture

PTH403 – Experiential Learning 0+10 8th Department of plant

(Yeast) pathology, School of
Agriculture

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 Commercial Aspects of Yeast

Yeast plays a crucial role in various commercial applications, primarily in the

food and beverage industry. Here are some of the key commercial aspects
related to yeast:

1. Baking:-
- Yeast is widely used in baking to leaven bread and other baked goods. It
ferments sugars in the dough, producing carbon dioxide, which causes the
dough to rise.

- Commercial bakers often use different types of yeast, such as active dry
yeast or instant yeast, depending on their specific production processes and
requirements.

2. Brewing:-
- Yeast is a key ingredient in the brewing of beer and other fermented
beverages. It converts sugars from malted grains into alcohol and carbon
dioxide during the fermentation process.

- Brewers may use different strains of yeast to achieve specific flavors,

aromas, and characteristics in their beer.

3. Wine Making:-
- In winemaking, yeast is essential for the fermentation of grape juice into
wine. Yeast converts sugars into alcohol and contributes to the flavor profile of
the final product.

- Winemakers may choose different yeast strains to influence the taste and
aroma of the wine.

4. Bioethanol Production:-
- Yeast is used in the production of bioethanol, a renewable fuel. It ferments
sugars derived from crops like corn or sugarcane into ethanol, which can be
used as a fuel additive or as a standalone biofuel.

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5. Nutritional Yeast:-
- Nutritional yeast is a deactivated form of yeast that is rich in nutrients,
including B-vitamins. It is often used as a food supplement and flavor enhancer,
especially in vegan and vegetarian diets.

6. Yeast Extracts:-
- Yeast extracts are used in the food industry to enhance flavors in various
products such as soups, sauces, and snacks. They contain compounds like
glutamic acid that contribute to a savory or umami taste.

7. Pharmaceuticals:-
- Yeast is used in the production of pharmaceuticals, including vaccines and
therapeutic proteins. The ease of genetic manipulation in certain yeast strains
makes them valuable for producing complex proteins for medical purposes.

8. Enzyme Production:-
- Yeast can be engineered to produce specific enzymes used in various
industrial processes, such as the production of biofuels, textiles, and detergents.

9. Research and Development:-

- Yeast, particularly Saccharomyces cerevisiae, is widely used as a model
organism in biological research. Its well-understood genetics and cellular
processes make it valuable for studying fundamental biological mechanisms.

Understanding the commercial aspects of yeast is essential for industries that

rely on its unique biological properties for various applications. Yeast's
versatility makes it a valuable resource in multiple sectors, contributing to the
production of diverse products.

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 Potential of the sector
The yeast sector has significant potential across various industries due to the
versatile nature of yeast and its unique biological properties. Here are some
aspects highlighting the potential of the yeast sector:

1. Growing Demand in the Food Industry:-

- The food industry continues to drive demand for yeast, especially in baking,
brewing, and the production of savory flavor enhancers. As global populations
rise and consumer preferences evolve, the demand for yeast in food applications
is likely to grow.

2. Expanding Beverage Market:-

- With the increasing popularity of craft beers, specialty wines, and other
fermented beverages, the yeast sector is poised to benefit from the expansion of
the beverage market. Brewers and winemakers are exploring different yeast
strains to create unique and diverse flavor profiles.

3. Biofuel Production:-
- As the world seeks sustainable energy sources, the demand for biofuels is
expected to rise. Yeast plays a crucial role in bioethanol production, and
advancements in yeast engineering may further enhance its efficiency in
converting biomass into renewable fuels.

4. Biotechnology Applications:-
- Yeast is widely used in biotechnological processes for the production of
enzymes, pharmaceuticals, and other valuable compounds. Continued
advancements in genetic engineering and synthetic biology are likely to expand
the range of applications for engineered yeast strains.

5. Health and Nutrition Products:-

- Nutritional yeast, known for its high vitamin B content, is gaining popularity
as a health and nutrition supplement. The demand for such products may
increase as consumers become more conscious of their dietary choices.

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6. Research and Development:-
- Yeast, particularly Saccharomyces cerevisiae, remains a crucial model
organism in biological research. Ongoing research and developments in genetics
and synthetic biology may lead to the discovery of new applications and
improvements in industrial processes involving yeast.

7. Pharmaceutical Production:-

- Yeast's role in pharmaceuticals, particularly in the production of vaccines

and therapeutic proteins, is likely to expand. The ease of genetic manipulation
in yeast makes it a valuable tool for the efficient production of complex
molecules.

8. Emerging Markets:-
- As emerging markets continue to develop and consumer preferences evolve,
there may be increased opportunities for yeast-related products in regions where
traditional fermentation-based foods and beverages become more popular.

9. Sustainability Focus:-
- The yeast sector is well-positioned to align with the growing emphasis on
sustainability. Yeast is a key player in sustainable practices, especially in the
context of biofuel production and other environmentally friendly processes.

10. Innovation and Technology Adoption:-

- Advances in fermentation technology, genetic engineering, and bio
processing are likely to drive innovation in the yeast sector. Companies that
embrace new technologies and methods may gain a competitive edge in the
market.

The potential of the yeast sector lies in its adaptability, versatility, and the
ongoing advancements in biotechnology. As industries continue to seek
sustainable and efficient solutions, yeast is expected to play a crucial role in
meeting these demands across various applications.

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 Activity Chart
The ELP programme “Propagation of Quality Planting Material in
Forestry Species”, was initiated under the aegis of Department of
Plant Pathology, School of Agriculture, Lovely Professional
University, Phagwara (Punjab). Unit was established during 2022-23
for production technology of yeast. The students work out bio control
of yeast Colletotrichum musae. We also do the purification of yeast
and also checking the biocontrol of Colletotrichum musae.
We also do the preparation of water agar media and inoculation of
yeast in sense of understanding its use as bio control agent.
The major activities are listed below:-
 Isolation of different strains of yeast (Method = Leaf disk
technique).
 Formulation for culture molasses urea
 Preparation of PDA (Potato Dextrose Agar) media and
inoculation of pathogen for maintaining culture.
 Preparation of PDAY media and inoculation of yeast and
pathogen in sense of understanding its use as bio control agent.
 Preparation of water agar media and inoculation of yeast in
sense of understanding its use as bio control agent (With banana
peels).

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 Activity Chart

ELP: Production technology for yeast (PTH - 403)

Coordinator: Dr. Deewakar Baral (UID)

Activity Schedule of ELP: Production technology for yeast

WEEK ACTIVITY
1st week Isolation of different strains of yeast by
leaf disk technique in YePDA media
2nd week Liquid formulation of molasses urea
3rd week  Liquid formulation of molasses
urea
 YePDA in petri-dishes for isolation
of yeast through leaf disk technique
4th week  Preparation of YePDB( Yeast
Extract Peptone Dextrose Broth)
 Streaking of yeast in YePDA media
 Mixing of grain
powders(Inoculated with yeast)
with YePDB
 Final purification of yeast in zig-
zag manner
5th week  Slant preparation
 Bio control of yeast Colletotrichum
musae(In water agar media)
6th week Preparation of water agar media and
inoculation of yeast in sense of
understanding its use as bio control
agent(Bio control of yeast Colletotrichum
musae)
7th week  Preparation of PDA (Potato
Dextrose Agar) media and
inoculation of yeast in sense of
understanding its use as bio control
agent.
 Transferring of colletotrichum to
new PDA media for getting pure
culture of Colletotrichum for future
use.
 Isolation of different strains of
yeast: - (Method = Leaf disk
technique).
 Preparing culture of
Colletotrichum for future use
 Growth of yeast from YePDB
media.

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8th week
 Purification of yeast through
dilution technique
 Isolation of pathogens
9th week
 Transfer and maintaining
culture of pathogen.

10th week
 Purification of yeast
 Evaluation of biocontrol
activity of yeast against
pathogens.
11th week  Cell count of yeast grown in
molasses urea
 Dry formulation of yeast
(yeast-rice cake)
12th week  Comparison of yeast growth
in different liquid media
13th week  Biocontrol activities of yeast
14th week  Biocontrol activity of yeast on
Colletotrichum gloesporioides
15th week  Biocontrol activity of yeast on
Sarocladium oryzae(Sheath
rot), Helmenthosporium
oryzae(Brown spot) ,
Ustilaginoidea virens(False
smut)
16th week  Biocontrol activity of yeast on
Colletotrichum musae

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 Summarized Activity Chart
Week -- WEEK 1 WEEK 2 WEEK 3 WEEK 4 WEEK 5
Month---
1ST month 1. Isolation of 1. Liquid 1. Liquid 1. Preparation of 1.Slant preparation
different strains formulation of formulation of YePDB (Yeast Bio control of yeast
of yeast by leaf molasses urea. molasses urea. Extract Peptone Colletotrichum musae (In
disk technique 2. YePDA in Dextrose Broth). water agar media).
in YePDA petri-dishes 2. Streaking of
media. for isolation of yeast in YePDA
yeast through media.
leaf disk 3. Mixing of grain
technique. powders
(Inoculated with
yeast) with
YePDB.
4.Final
purification of
yeast in zig-zag
manner.
2ND month 1. Preparation 1. Preparation of
1. Purification 1. Transfer and 1.Purification of yeast
of water agar PDA (Potato
media and Dextrose Agar) of yeast maintaining 2. Evaluation of
inoculation of media and through culture of biocontrol activity of
yeast in sense of inoculation of dilution pathogen. yeast against pathogens.
understanding yeast in sense of technique.
its use as bio understanding its 2. Isolation of
control use as bio control pathogens.
agent(Bio agent.
control of yeast 2. Transferring of
Colletotrichum colletotrichum to
musae). new PDA media for
getting pure culture
of Colletotrichum
for future use.
3. Isolation of
different strains
of yeast: -
(Method = Leaf
disk technique).
4.Preparing
culture of
Colletotrichum
for future use
5. Growth of
yeast from
YePDB media.
3RD month 1. Cell count of 1. Comparison of 1. Biocontrol 1. Biocontrol 1. Biocontrol activity of
yeast grown in yeast growth in activities of activity of yeast yeast on Sarocladium
molasses urea. different liquid yeast. on Colletotrichum oryzae (Sheath rot),
2. Dry media. gloesporioides. Helmenthosporium oryzae
formulation of (Brown spot),
yeast (yeast-rice Ustilaginoidea virens
cake). (False smut).
2. Biocontrol activity of
yeast on Colletotrichum
musae.

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 Module Photographs
WEEK ACTIVITY
1st week Isolation of different strains of yeast by
leaf disk technique in YePDA media
2nd week Liquid formulation of molasses urea
3rd week  Liquid formulation of molasses
urea
 YePDA in petri-dishes for isolation
of yeast through leaf disk technique
4th week  Preparation of YePDB( Yeast
Extract Peptone Dextrose Broth)
 Streaking of yeast in YePDA media
 Mixing of grain
powders(Inoculated with yeast)
with YePDB
 Final purification of yeast in zig-
zag manner

Glimpses of the activities done by the student:-

15
16
 5th week  Slant preparation
 Bio control of yeast Colletotrichum
musae(In water agar media)
6th week Preparation of water agar media and
inoculation of yeast in sense of
understanding its use as bio control
agent(Bio control of yeast Colletotrichum
musae)
7th week  Preparation of PDA (Potato
Dextrose Agar) media and
inoculation of yeast in sense of
understanding its use as bio control
agent.
 Transferring of colletotrichum to
new PDA media for getting pure
culture of Colletotrichum for future
use.
 Isolation of different strains of
yeast: - (Method = Leaf disk
technique).
 Preparing culture of
Colletotrichum for future use
 Growth of yeast from YePDB
media.

8th week
 Purification of yeast through
dilution technique
 Isolation of pathogens

Glimpses of the activities done by the student:-

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18
 Week Activity
th
8 week
 Purification of yeast through dilution technique
 Isolation of pathogens

9st week
 Transfer and maintaining culture of pathogen.

10nd week
 Purification of yeast
 Evaluation of biocontrol activity of yeast against pathogens.
rd
11 week  Cell count of yeast grown in molasses urea
 Dry formulation of yeast (yeast-rice cake)
12th week  Comparison of yeast growth in different liquid media

13st week  Biocontrol activities of yeast

14nd week
 Biocontrol activity of yeast on Colletotrichum gloesporioides

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Week: 8

Activity 1: Preparation of media (water agar and Yeast extract peptone dextrose
agar media)

Description: water agar media were used for the isolation of pathogen which
will provide platform for the growth of the pathogen by utilizing the nutrient of
its own host. While YEPDA (Yeast extract peptone dextrose agar media is
specific for the yeast to growth.

Step 1: Weighing

For water agar preparation, weigh 20g of Agar-Agar by using weighing balance
and dissolved in 1l of distilled water, similarly 20g of peptone 20g of dextrose
and 20g of agar was used in 1l of distilled water for preparation of YEPDA.

Step 2: Boiling

Above said media were pour in conical flask and boiled by stirring using hot
plate.

Step 3: Sterilization

Conical flasks were covered using cotton plug and sterilized in autoclave at 15
psi for at least 15 minutes.

Activity 2: Isolation of yeast by leaf disc technique.

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Description:
Yeasts are present in every phyllo sphere of the plant which can be utilized for
the isolation yeast. Same phyllo sphere may carry different species of yeast
which can be used advantageously for controlling different diseases of fruits
and vegetables.

Step1: Collection of tender leaves

Collected tender leaves from any plant in early morning or probably from
polyhouse.

Step2: Pour YEPDA media in sterilized petri-dish. Pour media in sterilized

petri dish and spread it uniformly in the petri-dish.

Step3: Inoculation of leaves

Cut all the leaves in small pieces and transfer to petri-dish after washing and
drying.

Step4: Incubation

Covered the petri-dish with paraffin tape and brown paper and then incubated
for 24 hours.

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Week 9:-

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Activity 1: Purification of yeast through dilution:

Step 1:First sterilized the conical flask and petri-dishes in autoclave at 15psi for
15 minutes.

Step 2:Collected green tender leaves

Step 3:Washed the leaves with sterilized distilled water

Step 4:Then chop the leaves in small pieces.

Step 5:Then fill the sterilized conical flask with 50 ml of distilled water and put
the chopped leaves in the conical flask.

Step 6:Shake the conical flask well with the help of a vortex shaker

Step 7:

 Preparation of YePDA (Yeast Extract Peptone Dextrose Agar) media-----

In a sterilized conical flask made 200 ml of YePDA by mixing 4.5 gm
yeast extract powder (60%) and 1.0 gm of agar-agar type I powder in 200
ml of distilled water, and then mix them well with a glass rod stirrer.
 Then cotton plug the cotton plug tightly and then wrap the mouth of
conical flask with a silver foil (In this case no need to tie the mouth with
thread) or cover the mouth with the brown paper and then tie it with the
fine white thread, then sterilize it in autoclave at 15psi pressure for 15
minutes.
 Add some pinches of streptomycin sulphate in the YePDA media which
works as an antibacterial agent, then pour the media in the Petri-dishes
and let them to solidify in the laminar air flow.

Step 8:Took a 1ml micropipette and with the help of it took 1ml from the
diluted media of leaves and distilled water and then transferred it to every petri-
dishes containing YePDA media.

Step 9:After some time wrap each Petri-dishes with the paraffin tapes.

Step 10:Then wrap the petri-dishes well with brown paper and tie it with fine
white thread.

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Step 11:Kept the petri dishes in incubator (26 °C to 30 °C) for better growth of
yeast.

Activity 2: Isolation of pathogens:

Step 1:First collected diseased sample of banana showing symptoms of
anthracnose, which is caused by a pathogen named Colletotrichum musae.

Step 2:Sterilized the conical flasks and petri-dishes in autoclave at 15psi

pressure for 15 minutes.

Step 3:

 Made water agar media of 100 ml in a conical flask by adding 2.5gm of

agar-agar type I powder in 100 ml of distilled water and stirred them well
with a glass rod stirrer. [water agar media were used for the isolation of
pathogen which will provide platform for the growth of the pathogen by
utilizing the nutrient of its own host].
 Cotton plugs the conical flask tightly and then wrap its mouth with a
silver foil, then autoclave it at 15psi pressure for 15 minutes, then pour
100 ml distilled water in another sterilized conical flask and then cotton
plug it and wrap it with a silver foil and also autoclave it at 15psi pressure
for 15 minutes.
 Add few pinches of streptomycin sulphate in the water media which acts
as an antibacterial agent.
 Put the water agar media in the petri-dishes and let them to solidify in the
laminar air flow.

Step 4:Took banana with typical black spots or having anthracnose symptoms
and with the help of a sterilized scalpel cut those black spots and put those in a
petri-dish containing distilled water (Poured the autoclaved distilled water),
after washing those pieces sterilize them partially in 70% ethanol (By mixing 3
ml distilled water with 7 ml of ethanol in a petri-dish) for few seconds. After
that dry them on a blotting paper.

Step 5: With the help of a sterilized L-shaped loop took those pieces and
transferred them on the solidified water agar media in petri-dishes.

Step 6: Then wrap the petri-dishes with the paraffin tapes and then wrap them
well with a blotting paper and tie it with a fine white thread.

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Step 7:Kept them in incubator for 24 to 48 hrs.

Step 8:After taking out of incubator white filamentous thread like structure was
observed.

Step 9:Removed the black spot pieces from the petri-dishes with the help of
sterilized needle.

Step 10:Again wrap those petri-dishes well and store them in incubator for
better growth of pathogen.

Step 11:After few days white mass of pathogen had been seen grow on every
petri-dish (i.e., Colletotrichum).

WEEK 10

Activity 1: Purification of yeast through dilution:

Step 1: First sterilized the conical flask and petri-dishes in autoclave at 15psi
for 15 minutes.

Step 2: Collected green tender leaves

Step 3: Washed the leaves with sterilized distilled water

Step 4: Then chop the leaves in small pieces.

Step 5: Then fill the sterilized conical flask with 50 ml of distilled water and
put the chopped leaves in the conical flask.

Step 6: Shake the conical flask well with the help of a vortex shaker

Step 7:

 Preparation of YePDA (Yeast Extract Peptone Dextrose Agar) media-----

In a sterilized conical flask made 200 ml of YePDA by mixing 4.5 gm
yeast extract powder (60%) and 1.0 gm of agar-agar type I powder in 200
ml of distilled water, and then mix them well with a glass rod stirrer.
 Then cotton plug the cotton plug tightly and then wrap the mouth of
conical flask with a silver foil (In this case no need to tie the mouth with
thread) or cover the mouth with the brown paper and then tie it with the

25
 fine white thread, then sterilize it in autoclave at 15psi pressure for 15
minutes.
 Add some pinches of streptomycin sulphate in the YePDA media which
works as an antibacterial agent, then pour the media in the Petri-dishes
and let them to solidify in the laminar air flow.

Step 8:Took a 1ml micropipette and with the help of it took 1ml from the
diluted media of leaves and distilled water and then transferred it to every petri-
dishes containing YePDA media.

Step 9:After some time wrap each Petri-dishes with the paraffin tapes.

Step 10:Then wrap the petri-dishes well with brown paper and tie it with fine
white thread.

Step 11:Kept the petri dishes in incubator (26 °C to 30 °C) for better growth of
yeast.

WEEK 11:-
Activity1: Viable cell count-
Description: By using viable cell count we were able to get the number of
viable cell of yeasts per ml of media.

Step 1: Sterilization

Petri dishes were sterilized in the Autoclave at 15 psi for 15 mins.

Step 2: Preparation of YEPDA Media and its sterilization

For this take a conical flask and add 200ml distilled water, 4.5gm yeast
extract powder and 0.5gm agar-agar type 1 powder and stir it well and keep
it in autoclave for sterilization. Also add Streptomycin sulphate in it to kill
any bacteria if present.

Steps 3:Collection of tender plant leaves

Tender plant leaves primarily from green house areas and shadow areas are
collected.

26
Step 4:In the laminar airflow pour media in the Petri dish and spread it evenly
and let it cool.

STEPS 5: Cut the leaves into small pieces and wash them in the distil water and
dry them using blotting paper.

Step 6:Put the pieces in the media in petri dish and wrap it with the paraffin
take covered with brown paper and keep in the incubator for 6hrs and 12hrs.

Step 7:In the time stated above remove the leaves. Yeast as minute creamy
white dots will appear. Again, keep it in incubator for proper growth of the
yeast.

Step 8: Now take new petri dishes sterilize them again and pour YEPDA media
in it and leave for solidifying.

Step 9: Meanwhile take test tube filled with 5ml distilled water.

Step 10: Transfer the yeast from the culture to the test tube using sterilized loop
and mix it well using vortex mixer.

Step 11: Now using sterilized loop take sample from the test tube and make a
streak in straight line on YEPDA media in petri dish and keep it in incubator
after covering with brow paper.

Step 12: As the growth of yeasts appear on straight line again prepare YEPDA
media, autoclave the petri dishes and in laminar air flow pour the YEPDA
media leave it to solidify.

Step 13: By sterilized loop take the sample of yeast growing in straight line and
mix it in 5ml distilled water and mix using vortex mixer to get an even
composition.

Step 14: Again, using a sterilized loop take the sample from test tube and form
a streak in zig zag style on the media and incubate it.

Step 15: After the growth has appeared in zig zag pattern again prepare YEPDA
media and sterilize it along with test tubes and in laminar airflow pour the
media in test tubes and leave for solidifying in slating position to increase the
surface area.

Step 16: From growth of yeast in zig zag pattern pick the sample using
sterilized loop and mix it in 5ml distil water using vortex mixer and continue

27
with making its streaking in zig zag pattern on slated media solidified in a test
tube and incubate it.

Step 17: As the growth appear now the time is for viable cell count of yeasts.

Step 18: For viable cell count take six petri-dishes and 12 test-tube which was
previously sterilized and divide each petri-dishes in 6 sections equally with the
help of a marker.

Step 19: Pour 9 ml distilled water in each test tubes and sterilize the test tubes
in autoclave.

Step 20: From yeast culture in slanting test tubes take the yeast sample by
sterilized loop and transfer it to the test tubes containing 9ml distilled water and
mix with vortex mixer to get a diluted solution of yeast.

Step 21: Repeat this process with the rest of the test tubes.

Step 22: Take the set off each six test-tubes for serial dilution process and for
that purpose took 1 ml micropipette and then with the help of it take 1ml from
the 1st test tube and then transfer it to 2nd test-tube and repeat this process till the
6th test tube.

Step 23: with the help of 1microliter micropipette take the diluted yeast from
the 6th test tube and then pour 6 to 8 drops in the marked 6 th position of first
three petri-dishes. Then repeat the same process till the first test tube.

Step 24: After all these functions wrap the petri-dishes with the paraffin tapes
and then cover them with the butter paper and then tie them with fine white
thread.

Step 25: Keep these petri-dishes in incubator under standard condition of 15atm
pressure for 12 to 24 hours.

Step 26: After few days petri-dishes were taken out and cell counting was done
using digital colony counter.

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29
Week12:-

Activity1: Cell count of yeast grown in molasses urea

Step1: Serial Dilution of yeast grown in molasses urea

Transfer yeast grown in molasses urea with the help of 1 ml micropipette to

glass tube containing 9 ml distilled water and shake it well on vortex mixer for
even distribution of cells.

Take 1 ml from that glass tube and transfer it to another tube containing 9 ml
distilled water and repeat this procedure up to 6 times.

30
Step 2: Marking of petri-dish

Take sterilized petri-dish and divide it in 6 compartments and mark 1 to 6 on

them with marker.

Step3: Pouring media in petri-dish.

Fill YEPDA in it and place for solidification.

Step4: Transferring of yeast cells from dilution to media

After solidification, take 10 micro litres from first glass tube with the help of
micropipette and drop 6 drops in particular compartment and repeat same with
all 6-glass tube.

1) Do this procedure for all solutions (T1 to T11).

Activity 2: Dry formulation of yeast:-

1) Dry formulation of yeast (yeast-rice cake):

Step 1: Grinding of rice floor
Step2: Sterilization
Then sterilize it within autoclave at standard conditions that are 15 psi at
least for 15 minutes.
Step3: Kneading of rice flour
Mix it with molasses to form a dough shaped cake.

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 Step4: Inoculation of yeast
Then take 1 ml Yeast containing YEPDB with the help of micropipette
and inoculate yeast cake.

Step5: Fermentation

Allow it to ferment for 3 days in incubator.

Step6: Cell count

After fermentation perform cell count for yeast cake by taking 1 gm yeast
cake from it and add in glass tube containing 10 ml distilled water and
serial dilute it up to 6 time after mixing well it on vortex mixer.

Week 13:-
Activity1 :- Preparation of different media:-

 Step 1:- First prepared 1000 ml molasses urea by adding 15gm yeast
extract powder, 20gm molasses and 5-6 grains urea ball to 1000 ml of
distilled water.
 Step 2:- Then prepared YEPDB media by adding 2gm peptone, 2gm
dextrose, 1gm yeast extract powder in 100 ml distilled water.
 Step 3:- Next, Inoculate 1 ml of yeast from Yeast culture YEPDB in
every bottle.
 Step 4:- Incubated it for 4 days.

Activity2 :- Inoculation of yeast:-

For inoculation of yeast used leaf disk technique.

 Step 1: collect tender green leaves within 6 hours (preferred leaves from
green house), and sterilized conical flask and petri dishes in autoclave at
15psi pressure for at least 15min.
 Step 2: Preparation of YePDA (Yeast Extract Peptone Dextrose Agar)
media----- In a sterilized conical flask made 200 ml of YePDA by
mixing 4.5 gm yeast extract powder (60%) and 1.0 gm of agar-agar type
I powder in 200 ml of distilled water, and then mix them well with a
glass rod stirre.

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 Step 3: Then cotton plug the cotton plug tightly and then wrap the mouth
of conical flask with a silver foil (In this case no need to tie the mouth
with thread) or cover the mouth with the brown paper and then tie it with
the fine white thread, then sterilize it in autoclave at 15psi pressure for 15
minutes.
Step 4: Add some pinches of streptomycin sulphate in the YePDA media
which works as an antibacterial agent, then pour the media in the Petri-
dishes and let them to solidify in the laminar air flow.
 Step 5: Then, autoclave the YEPDA solution under 15 lbs pressure at
121 degree centigrade for 30 minutes.
 Step 6: Then, cut those leaves in small pieces, and wash them with
distilled water in a separate Petri dish.
 Step 7: With the help of sterilized L shaped loop transfer those leaf
pieces on YEPDA media in Petri-plate.

 Step 9: Then put all those Petri dishes wrapped with butter paper and
placed in BOD incubator for incubation for 12 hours.

 Step 10: After that, remove leaves with in respective 6 and 12 hours
respectively for maximum growth.

 Step 11: After removing leaves black dots can be seen.

 Step 12: After removing leaves again incubate for 12 hours.

 Step 13: Then take one white dot and mix it with 5 ml distilled water
with the help of vortex mixture.

 Step 14: After that take the solution and do streaking on another PDA
solution. (For isolation of single colonies and decreases competition
between different strains of yeast from mixed species or from the same
species).

 Step 15: Continue this streaking in same method until the isolation of
colonies of yeast, and its growth.

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Week 14:-
Activity 3: Comparing the growth in different media:-

 Step 1:- First we prepared 1000 ml of molasses urea by adding 15gm of

yeast extract powder, 20gm molasses and 5-6 grains urea ball to 1000ml
of distilled water.
 Step 2: Then prepared YEPDB media by adding 2gm peptone, 2gm
dextrose, and 1 gm yeast extract powder in 100 ml distilled water.
 Step 3: Then cotton plug the cotton plug tightly and then wrap the mouth
of conical flask with a silver foil (In this case no need to tie the mouth
with thread) or cover the mouth with the brown paper and then tie it with
the fine white thread, then sterilize it in autoclave at 15psi pressure for at
least 15 minutes.
 Step 4: Inoculate 1 ml of yeast with the help of 1ml micropipette from
Yeast culture YEPDB in every bottle.
 Step 5: Incubate it for 4 days.
 Step 6 : It had been observed that growth of the yeast had came first in
molasses urea than YEPDA.

34
 List of Students
S.No. Name Registration no. Contact number

1. Loveneesh 12010316 9053450415

2. Ritabrata Das 12013144 8170073879
3. Anuran Roy 12010742 9889913255
4. Souhardya Adhikari 12228200 7718749996
5. Tsering Dondu Ngangmu 12010608 8257072682
6. Biswajit Mondal 12000670 9800394050
7. Manoj Sorout 12001734 9992254818
8. Mansha Makhija 12001316 8077723595
9. Anshmaan Kaur 12002892 7470482923
10. Chinmoy Goswami 12002539 8967013802
11. Radha Gara 11916499 8905157894
12. Samyajyoti 12018203 9475648706
Bandyopadhyay
13. Priti Rani 12012161 7061634769
14. Sukhamritpaul Singh 12012154 9855027560
15. Sunav Raj Kashyap 12014234 6000574132
16. Gaikodi Abhiram 12010883 9014278130
17. Biswapriya 12003153 9933111811
Roychoudhury
18. Paawan Popli 12019979 9466904879
19. Patan Khadar Basha 12014854 9133274933
20. Shwetank Jha 12001522 6200749343

35
21. Vankayalapati Vyshnavi 12018988
22. Gandham Lakshmi 12019484 9550913467
Chaitrika
23. Abijith Manoj 12009229 7591923132
24. Sarithala Kullayappa 12002956 6304119079

36
 Conclusion
Experiences gained during ELP by students:-
 ELP programme was an exposure programme for us, in which theoretical
knowledge was applied in real field conditions.
 The program was efficient enough to developed entrepreneurship quality
among us.
 The program incorporated team work and coordination among the
students to complete the assigned task.
 The program also helps in creating relationships between the students and
the stakeholders.

Feedback
• This Experiential Learning Programme (ELP) really helped us to know about
real life field problem, farm situation and obstacles that growers face during the
propagation and marketing of produce.

• ELP programme helped us to understand about new techniques to create

venture for selfemployment.

• ELP programme helped us to change the mindset from normal thinking to

innovative thinking & create our own business-oriented skills.

Submitted By:-
1. Loveneesh 12010316 9053450415
2. Ritabrata Das 12013144 8170073879
3. Anuran Roy 12010742 9889913255
4. Souhardya Adhikari 12228200 7718749996

Dr. Deewakar Baral

Coordinator HOD, Horticulture HOS,
School of Agriculture

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