BUGEMA UNIVERSITY

SCHOOL OF NATURAL SCIENCES

DEPARTMENT OF AGRICULTURE SCIENCES

FIELD REPPORT SUBMITTED IN PARTIAL FULFILLMENT FOR A

BACHELOR OF SCIENCE IN AGRICULTURE WITH CROP SCIENCE
AND MANAGEMENT

BGCP 411 / 421: GENERAL PLANT PATHOLOGY

BY:

MAGALA NICODEMUS

17/CSM/BU/R/0010

TO:

MADAM.NAFULA RACHEAL

DECEMBER, 2021

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AFEILD TRIP REPORT ON PLANT PATHOLOGY CARRIED OUT AT NATIONAL
CROPS RESOURCES RESEARCH INSTITUTE (NACRRI), NAMULONGE ON 6TH
BDECEMBER 2021

ACTIVITIES CARRIED OUT DURING THE NAMULONGE RESEARCH FILED TRIP

CARRIED
Introduction

At our arrival at Namulonge research center, we were introduced to different bio

technicians, introduced ourselves.futher more we were introduced to Mr. KANYESIGYE
DALTON, the bioscience laboratory who introduced us the activities we were involved in during
the field trip period at National Crops Resources Research Institute (NaCRRI) Namulonge on 6th
December, 2021

Program of activities we observed and done

During the period at Namulonge research institution, we were taken through the following
activities by Mr.KANYESIGYE DALTON the bio technician

1. Introduction to different laboratories for example,

 Tissue culture lab

 Conservation lab
 Molecular la

2. Diseases detection

 Bacterial infection.
 Viral infection
 Fungal infection

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Disease detection

Before identifying which kind of pathogeny, genetics has to be identified, here different activities
are carried out,

Isolation: isolation is done in order to analyze and identify whether the pathogeny is RNA or DNA
pathogeny (what is it?), cassava mosaic, brown strip or other diseases, for instance

Cassava mosaic disease: it is DNA viral disease

Cassava Brown streak disease: it is RNA viral disease

Note: DNA is stable therefore it is better for analysis than RNA

Quantification: involves quantifying the amount and quality of DNA or RNA

We looked at RNA as being a single stranded

If it deals with,

RNA identification, polymerase chain reaction running is carried out (PCR)

DNA identification, BCNA is used to run it

Diseases breaking: after identifying the disease protein, to avoid it multiplying more and
affection the whole plant or other plants, the MRNA is broken down or cut off in order to stop
protein synthesis, how?, translocation is cut off completely . here, RNA 1 Technology is used to
break down MRNA to DNA

LABARATORY TOOLS

Some of the tools used during the experiments are:

 Water bath incubator  Motor

 Dry bath incubator  piston
 Fume hood  Concentrator
 Tissulizor  PCR
 Centrifuge

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Usages

PCR

Before we started the extraction, we were moved around the laboratory, in there, we were
shown the PCR which is used in the automatic identification of viruses, we were told by our
instructor that it can also be used to identify COLONA virus

Tissulizor

It is used in breaking down the cells and crushing the tissue down, it carries only 24 samples
per crushing and it is set for 1 minute

Motor and piston

If the tissulizer is not available, we put the cassava leaf in a motor and crushed it manually
with a piston. This was carried out in a fume hood, during the crushing, we added a little amount
of baffer for easy crushing, and baffer makes the sample stable

Reason for using a fume hood

This is to avoid the irritating effect that may be caused in the laboratory during the
experimentation activities

Note: water is not used during the crushing because it is not safe

Dry incubator and water bath incubators

After the crushing and addition of baffer, the sample was added in the cube, and numbered
well where then were taken for incubation

Procedures

 Water bath incubator: it uses water for heating the extracted tissue in cubes set at 65 0 c.
 Cubes containing the sample are inserted in troughs and covered, temperature is set and
wait for 30 minutes, but shaking is required for every after 10 minutes to avoid settling of
the tissue crushed.

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  Dry bath incubator: it uses only steam which heat the material, shaking is required to avoid
settling of the materials
 After the crushing, incubation is done either using the dry or water bath incubator
 The cubes are used for carrying the samples which are 1.5mls
 The cubes were first numbered according to the type of plant, variety, A=1, B= 2and
sample given name for example, (1, 2, and 3)
 The sample was crushed mixed with buffer
 After the crushing, the crushed sample was poured in tubes and taken for incubation for
30-65 minutes

During the incubation, the sample has to be shaken for ten minutes in order to have a uniform
sample

Centrifuge

 After the incubation process, the sample was taken in the centrifuge, here there was
separation of chlorophyll, proteins, acids (nucleic acid
 After separating and getting the two phases that is to say, clear up and not clear down due
to heaviness of the tissues they settled at the bottom of the tube
 The clear, upper sample is extracted using an extractor and poured into another tube.
 Isopropanol acetyl trim ethyl ammonium bromide (CTAB)

NOTE: it is the main component for the baffer

Isopropanol is added in order to precipitate to get a nucleic acid and in order to get a palate

PATHOLOGY SECTION (LEGUME SECTION)

 After the bioscience laboratory, we went to the pathology laboratory where identification
of bean diseases are carried out

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  We were taken through disease identification by Madam JUSTINE who had a sweet voice,
it was wahoo, I was hungry but it stopped immediately after hearing her sweet voice (by
the way)

ACTIVITIES CARRIED OUT

Listing fungal diseases symptoms

Listing viral diseases
Listing bacterial diseases

Fungal disease signs:

 Leaf rust (common leaf rust in corn)

 Stem rust (wheat stem rust)
 Sclerotinia (white mold)
 Powdery mildew

Fungal disease symptoms:

 Birds-eye spot on berries (anthracnose)

 Damping off of seedlings (phytophthora)
 Leaf spot (septoria brown spot)
 Chlorosis (yellowing of leaves)

Bacterial disease signs (difficult to observe, but can include):

 Bacterial ooze
 Water-soaked lesions
 Bacterial streaming in water from a cut stem

Bacterial disease symptoms:

 Leaf spot with yellow halo

 Fruit spot

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  Canker
 Crown gall
 Shepard’s crook stem ends on woody plants

Viral disease symptoms:

 Mosaic leaf pattern

 Crinkled leaves
 Yellowed leaves
 Plant stunting
Here we were told to search more from internet
Were looked at some diseases that attach beans
Root rot disease
Southern blight (sclerotium Rolfsii)
Fusurium root rot (fusiurum oxyosporum solania

Identification of diseases using alight microscope

Identification of disease using a bean leaf as specimen

Procedures

We Connected the light microscope to a power source

Turn the revolving nosepiece so the lowest objective lens is in position.
Mounted the specimen onto the stage. But before doing so, we looked for a leaf with clear
signs and symptoms
We used the metal clips to keep your slide in place. Make sure the specimen is positioned
in the center, right under the lowest objective lens.
Look into the eyepiece and slowly rotated the coarse adjustment knob to bring the specimen
to focus. Seeing that the slide was not touching the lens.
We adjusted the condenser for the maximum amount of light.

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 Now we slowly rotated the fine adjustment knob until we obtained a clearer image of our
specimen.
Now we examined our specimen.
We applied agel at the specimen, where we sow tree like threads with heads symbolizing
the presence of the disease

Culturing the disease

We concluded by studding how to grow the disease, where by a spore is picked from the
plant and placed in a growth media, which contain the food which the disease eat

Food is specific for every spore, they are placed in petrel dishes, white in color

The specimen is placed in cool temperatures in fridge since it favors their growth

We were all asked to observe the specimen in the microscope

Conclusion

The field trip was successful; the research center had good facilities that enabled me
participate practically in all the activities carried out; the supervisors/ trainers of the different
sections who taken us through were not selfish with their knowledge, ideas and experience. They
were willing to teach and converse with us and they took us through all I was meant to learn.

Am glad to have passed visited Namulonge research institute and I promise to use the knowledge
learnt where applicable.

Challenges I faced

The challenges I encountered during my trip included;

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  It was difficult to keep time as for launch
 Dusty roads
 Hunger

Recommendation

 To Namulonge research institute

I recommend Namulonge research institute to;

 Improve on infrastructure such as roads, renovation of old laboratories

 Buy new over coat for laboratory technicians
 Buy uniforms to workers depending on their section for smartness and identification.
 Build a simple restaurant where to get food
 Provide food to visitors like us

 To Bugema University:

I recommend Bugema University to;

 Put more emphasis on practical work

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Reference

https://www.canr.msu.edu/news/signs_and_symptoms_of_plant_disease_is_it_fungal_viral_or_b
acterial