Introduction.
- Banana is the single most important staple crop in Uganda contributing about 30 percent of total food consumption and 14 percent of total crop value.
- Banana cultivars include: Mpologoma, Musakala, Mbwazirume, Nfuuka, Nakinyika, Nakabululu and kisansa, Kabula and other local varieties.
- Pests and Diseases are among the most important factors in banana production worldwide. They are the reasons for which all of the world’s breeding programs were created and remain a primary focus of all current programs.
- Recently, diseases also became principal targets of biotechnological efforts to improve this crop.
Objectives.
- Micro-propagation of banana genotypes.
- Production of virus free banana seedlings.
- In-vitro and filed screening of banana crops in wilt infested areas.
- Natural selection of banana crops in Banana Bacterial wilt infested areas.
Preparation of the stock solutions
As a prerequisite to the media preparation, stock solutions (MS-A, -B, -C and -D) of known concentration are prepared by weighing required quantities of MS salts utilizing a digital analytical weighing balance and dissolving them in MilliporeTM water.
Micropropagation of Banana through organogenesis.
- Organogenesis is the formation of individual organs such as shoots and roots either through callus formation or emergence of adventitious organs directly from explants.
- The four stages in micropropagation are:
Stage 1: Initiation of sterile culture and establishment
Stage 2: In-vitro callus induction, shoot proliferation and multiplication
Stage 3: In-vitro rooting
Stage 4: Ex vitro establishment
Stage 1: Initiation of sterile culture and establishment
Selecting an explant
- Should have 5 photosynthetically active leaves and inter foliar space must be not less than 5.0 cm, 25-30 active roots at the end of secondary hardening stage.
- Should be free from any visual symptoms of leaf spot, pseudo stem rot and physical deformations.
Preparation of explants
- Wash the suckers thoroughly in tap water, remove roots and leaf sheaths, cut and trim basal portion of the corm to a size of 12*12*15 mm.
- Soak them in sodium hypochlorite (detergent) mixed with tween twenty for 30 minutes and shake
- continuously, Wash with distilled water to remove the detergent particles.
- Transfer them to laminar air flow chamber for further sterilization process.
Stage 2: In-vitro callus induction, shoot proliferation and multiplication
- Transfer of explant to tissue culture medium, Seal properly, label the tubes or bottles with date, name of the variety and the person in charge and transfer to the growth chamber.
Sub-culturing.
- Properly clean off the phenolics and old parts from tissues that successfully grown inside the Laminar air flow, cut the tissues into two (longitudinally), Subculture each new part into fresh growth media with similar composition.
Shoot proliferation
- Transfer the plants to shooting media to initiate shoot development.
- MS medium supplemented with the different concentrations and combinations of Kinetin, BAP for multiple shoot regeneration.
- This stage lasts for two weeks and the sole purpose is for the explants to develop shoots.
Stage 3: In-vitro rooting
- Transfer fully developed shoots to rooting media to initiate root development.
- MS medium supplemented with the different concentrations and combinations of rooting media like NAA, IBA for root regeneration.
Stage 4: Ex vitro establishment
Acclimatization
- Remove the in vitro regenerated plantlets from the culture vessels, clean the roots gently with running tap water to remove the adhering agar, transfer to the green house.

