Laboratory Visit in Institute of Plant Breeding (IPB)

Learning Activity 5 of AGRI 42

TESORO, Gabriel Paul A.; Y2-2L; FIC: Ms. Clare Hazel R. Tabernilla

A. Materials needed for mass production

1. Infected banana plant
2. Healthy banana plant
3. Greenhouse
4. Aphid container
5. Equipment for transferring aphids
6. Cage for safe exposure of the banana plant to inoculants
B. Target pest of the selected Biocon agent
• Target pathogen pest – Banana bunchy top virus (BBTV): Viral Pathogen.
C. Mass rearing procedure of the selected Biocon agent
• The researchers should first know the banana specie that may have resistance against
BBTV.
• This way, the researchers will know what species should be cultivated in resistance to
BBTV.
• More so, Sir Jayvee Mendoza said that the IPB needs help in producing species
of Musa balbisiana through tissue culture. In the meantime, they plant it with seeds.
However, they are currently making protocols for its tissue culture procedure for it to
be grown for further experiments.
D. Procedure in field release of the Biocon agent
• A researcher should know the aspects of the disease triangle. This way, they would
know what procedure to follow.
• Rearing and forming viruliferous Pentalonia nigronervosa (aphids).
o Sir Jayvee rears the aphids in an aphid container.
o He will starve the aphids inside the container for 4 hours to use them immediately
for inoculation because they are hungry.
o He will put the aphids in the infected plant for four hours or overnight for virus
acquisition before transferring them to a healthy banana plant.
• Inoculation of aphids to the plants
 o After forming the viruliferous aphids (virus-carrying aphids), a researcher can
transfer them to a test plant, and a positive control plant using the equipment for
transferring aphids (see Figure 1).
▪ Positive control plants are the plant species that we susceptible to the
disease. Lakatan banana (Musa acuminata) is the positive control plant of
banana species (see Figure 4).
▪ Test plants are those that researchers will test with resistance.
o The positive control plant will be the indicator that the inoculation is successful. If
it does not show any signs or symptoms, the researcher will have to do the
procedures from the start.
• Resistance test of different species of the host to the virus strain
o After inoculation, they will remove the aphids from the plant and isolate them in a
greenhouse to observe the virus strain.
▪ Note: The inoculation is used for the different banana varieties to know
those resistant from not.
o Sir Jayvee will observe the Musa sp. (see Figure 3) if it will be resistant against
BBTV by getting sample from its leaf (see Figure 2).
• Release to the field
o From the greenhouse, they will now put the resistant varieties in the field for trials
in natural conditions.
o It will let the researcher know if the variety is resistant if planted in the field.

Figure 1. Sir Jayvee transferred aphids Figure 2. The spots are the samples that Sir
(inoculants) from the infected plant to the Jayvee got to know if the leaf has the presence
healthy one. of BBTV.
 Figure 3. Musa sp. is observed in the
greenhouse for its resistance test.

Figure 4. Musa acuminata (Lakatan banana)

plant