ANNEX 23 - MYCOSPHAERELLA SPECIES TRAINING at THE NELSON MANDELA-AFRICAN INSTITUTE OF SCIENCE AND TECHNOLOGY

Report on the training on isolating Mycosphaerella species from banana leaves to the East Africa laboratory banana team

Amos Alakonya, IITA Plant Pathologist, Onne, Nigeria

The training and analyses were held at the Nelson Mandela-African institute of Science and Technology (NM-AIST), Arusha from 26th September to 1st October 2016 and were conducted by Amos Alakonya Banana Plant Pathologist IITA-Onne. The trainees were Janet Njeri (PhD Research Fellow), Kennedy Jomanga (IITA-Arusha) and Evans Were (IITA Kawanda).

Objective of the trainingThe objective of this training was to provide hands on experience in isolation of Mycosphaerella spp. from banana leaf samples. Further, it also served as an avenue to identify gaps that need attention within the Sigatoka project

The following were the activities covered/achieved during the training.

Identification and sampling of Sigatoka infected leafs. Glassware, media and laboratory accessory sterilization. Media preparation. Sample processing. Pre-culture leaf examination under microscope. Culture of infected leaf samples. Microscopic examination and identification of Mycospaerella spp. germinating/viable ascospores

and conidia. Aseptic picking, culture and incubation of Mycospaerella spp. germinating/viable ascospores and

conidia. Photographic laboratory data capture during Mycospaerella spp. isolation.

Figure 1: During the training session the participant L-R Evans Were (IITA Kawanda), Janet Njeri (PhD Student) and Kennedy Jomanga (IITA-Arusha) being guided by Amos Alakonya. A) Amos guides the participants on how to locate lesions on banana leaves with intact pseudothecia under the stereomicroscope for isolation Mycosphaerella spp. B) each participant clearly identified such lesions and C) the lesions were clearly marked out and carefully excised from the leaf samples.

Results of the training

Ascospores and conidia discharged onto water agar Petri plates were observed under the stereo

microscope. Some of these structures were seen germinating. Each participant was able to observe these

structures and practiced their picking using a needle and scalpel. The participants were also taught on how

to distinguish similar structures from other fungi especially the epiphytes or saprophytes such as

Alternaria alternata.

Figure 2: Conidia and ascospores discharged onto water agar and Petri plates observed under the stereo microscope. A) A conidium germinating (shown by black arrow) and B) ascospores (shown by white arrows)

Participant feedback after the trainingAlthough the training was intense due to the limited time, the participants had time to ask all the necessary questions relating to the isolation procedure. The three participants confidently agreed that the training was very necessary and had met their expectations. They had gained the fundamental techniques and confidence for isolating Mycosphaerella spp. a procedure they had previously struggled with. I can confidently conclude that the participants will be able to undertake all the isolation procedures of Mycosphaerella spp. hereafter and therefore the objective of the training was attained.

Challenges and recommended actionsThe IITA pathology unit at NM-AIST lacks a +4°C fridge, which is indispensible for storage of V-8 culture media and unused agar plates. Microscope had low magnification, a better microscope is recommended with at least X50 objective magnification is recommended. Acquiring the equipment can speed up the isolation work.

Trip was concluded with a visit to Mr. Jomanga’s family house where we enjoyed good Tanzania food.

We left Arusha for Kenya and Uganda on 1st October 2016.