THE STATUS OF WILT ( pv musacearum) IN WESTERN 1 J.N. Mbaka, 2 M. Mwangi, 2 V.G. Nakato, 3J.Auma and 4B. Odero

1Kenya Agricultural research Institute (KARI) National Horticultural Research Centre, Thika P.O. Box 220-01000, Thika, Kenya Tel. +254 67 24332 E-Mail [email protected] 2 International Institute for Tropical Agriculture (IITA) P.O Box 7878 Kampala, Tel +256 41 223445, 221009 Fax: +256 41 223494, 220217 Email: [email protected]:[email protected] 3 Rural Energy and Food Security Organisation (REFSO) P.O.Box 342, Busia, Kenya 4Catholic Relief Services (CRS) P.O. Box 49675-00100, GPO, Nairobi, Kenya Email: [email protected] Tel: 0733 401401 Mode of presentation in the KARI Conference 2008: ORAL Author for correspondence Jesca Njeri Mbaka National Horticultural Research Centre P.O. Box 220-0100, Thika, Kenya Email: [email protected] Tel: + 254 722 88 24 22

Mbaka et al Status of banana Xanthomonas wilt2 in Kenya-

ABSTRACT

Banana Xanthomonas Wilt (BXW), caused by the bacterium Xanthomonas campesris pv

musacearum was reported in a single farm in Mukono district of Uganda in 2001. By

early 2006 it was confirmed in 33 districts of Uganda, parts of Democratic Republic of

Congo, and . Disease spread between plants is mainly by insect

pollinators and contaminated farm tools. The main mode of long distance spread is

banana trade (, leaves and planting material). Insect transmitted infections lead to

wilting and rotting of the banana male , pre-mature ripening and rotting of fingers

that become unpalatable. Infections from contaminated tools and infected suckers start as

wilting of leaves that eventually dry. Infected plants die within two weeks. Yield losses

can be up to 100%. Due to its rapid spread, BXW poses a great threat to banana

production in the East and Central Africa region. Understanding of the disease status and

establishment of farmer practices that may pre-dispose to BXW are crucial to

preparedness for effective disease management. The objectives of this study therefore

were to establish, the status of BXW and the existing banana production practices in

banana orchards in Western Kenya. A survey was conducted in September, 2006 in 14

districts located in Western and Nyanza Provinces. A questionnaire was administered by

face to face interview with the farmers to capture data on banana production practices.

Wilted bananas were observed in Teso, Bungoma and Busia districts in western Kenya.

Symptoms were similar to those previously reported in Uganda. Pathogenicity tests

confirmed the causal organism as Xanthomonas campestris pv mucasearum. The

outbreak in western Kenya is significant because this region produces over 60% of

bananas in Kenya, and therefore could have serious consequences on food and income

2 Mbaka et al Status of banana Xanthomonas wilt3 in Kenya-

security of small-scale banana farmers. Technologies developed and validated in Uganda

can be used to stop disease spread.

Key words: bacterial, banana, control, disease

INTRODUCTION

Banana (Musa spp. L) is the fourth most important global food crop after rice, wheat and in terms of gross value of production (FAOSTAT, 2003). About 20 million people in the East and Central Africa region depend on banana for food. A wide range of genetic diversity of bananas is found in different areas of Kenya. Factors such as local tastes, eating habits, market demand and environmental conditions tend to influence their distribution (Nguthi, 1998). Diffusion, dissemination and adoption of the tissue culture technology aimed to address banana production constraints (pests, diseases and lack of adequate clean planting materials) has led to increased yields, higher demand for planting material and commercialization of tissue culture laboratories (Kahangi, 2003). By end of year 2006, Kenya had 82,518 hectares under bananas from this 505,258 metric tones valued at nearly 4 billion Kenya shillings were produced (MoA, 2006).

However, the recent spread of banana Xanthomonas wilt (BXW), since its report in

Uganda in 2001 (Tushemereirwe et al., 2003) is a big threat to banana production in The disease is caused by the bacterium Xanthomonas campestris pv. musacearum (Xcm) that was for long considered a problem of enset ( (Welw) Cheesman), in

Ethiopia (Thwaites et al., 2000). Enset is a Musa species mainly grown in for its importance for extraction of industrial starch from its corm. The first outbreaks outside

Ethiopia were reported in a single farm in Mukono district of Uganda in October, 2001

3 Mbaka et al Status of banana Xanthomonas wilt4 in Kenya-

(Tushemereirwe et al., 2003). By October 2003, the disease was confirmed in 10 more districts and is in 33 districts by early 2006 (Tushemereirwe and Kubiriba, 2006). Outside

Uganda, the disease was first reported in North Kivu province of the Democratic

Republic of Congo (DRC) in 2003 (Ndungo et al., 2004, 2006), Rwanda in 2004 and

Tanzania in 2005 (Mgenzi et al., 2006).

Many sources of infection are known or suspected for BXW including standing infected plants, plant residues, contaminated tools, contaminated and water, traded products

(fruits, leaves and planting materials). Observations at advancing disease fronts in

Uganda suggest that transmission to the male bud is the primary means of spread by pollinators (Eden-Green, 2004). Banana become most vulnerable to BXW infection at flowering stage since the pathogen can be transmitted between plants by insects that visit flowers for pollen and . Infection through the flower leads to rotting and total destruction of fruits, as well as wilting of the entire foliage. All banana cultivars are susceptible but some cultivars escape insect transmitted disease due to their floral morphology (persistent ). The most susceptible is Pisang Awak

(Tushemereirwe et al., 2003). Banana Xanthomonas wilt containment depends on two key actions: prompt removal of sources of inoculum and reducing and eliminating opportunities for spread. Establishment of diseases status and identification of pre - disposing factors are the initial steps in tailoring responses for management of the particular disease. The objective of this study therefore was to ascertain the status of

BXW and factors pre-disposing bananas to the disease in Western Kenya region. It was hoped that effective disease management strategies developed and evaluated in Uganda

4 Mbaka et al Status of banana Xanthomonas wilt5 in Kenya- would be applied in Kenya to stop disease spread to new areas and to reduce levels of severity in any affected areas.

MATERIALS AND METHODS

Disease surveys

A banana disease survey was carried out in Western Kenya region in August and

September 2006. The region was targeted due to its close proximity to Uganda, the banana trade and unrestricted movement of banana materials across the porous boundaries. The survey team was composed of two plant pathologists, one from Kenya

Agricultural Research Institute and one from the International Institute for Tropical

Agriculture, Kampala, Uganda and a horticulturalist from Rural Energy and Food

Security Organization, Busia, Kenya. The target areas were selected districts in Western and Nyanza provinces. A questionnaire was designed to capture data on banana cultivars grown, disease incidence and prevalence, soil types, banana production practices, banana marketing systems and utilization. The exact locations of the survey areas and sample collection sites were marked by use of a Geographic Positioning Satellite (GPS) instrument Magnum ®. The survey team visited the District Agricultural Officers

(DAOS) of the respective districts for desk top information on banana production. In every district, two main banana growing divisions were selected. A stratified random sampling was used to select 15 farmers from each division. A stratum was taken to be a farmer with at least 30 banana mats in production. The questionnaire was administered through face to face interview with the farmers. Thirty mats in each farm were scored for diseases and pests by the enumerators. Clear pictures of symptoms (diseases, pests and

5 Mbaka et al Status of banana Xanthomonas wilt6 in Kenya- physiological problems) of banana laminated and filed were used as visual aids for field diagnostics. A total of 365 farms were visited and respective farmers interviewed.

Isolation of Xcm on synthetic growth medium

Banana samples (pseudostem and fruits) suspected to have Xcm were put in polythene paper bags, sealed and put in a cold box. In the laboratory, the samples were thoroughly cleaned under running tap water and surface sterilized by dipping in 5 % sodium hypochlorite for 2 minutes, rinsed in distilled water and blotter dried. Isolation of Xcm was done on Yeast Peptone Glucose Agar (YPGA). The medium was poured onto 90 mm

Petri dishes and allowed to cool overnight. The following day, were picked from the infected samples with a sharp needle and plated on the medium. The plates were incubated at 25 0C for 72 hours. To obtain pure colonies, isolation was done on

Cephalexin Cellebiose Agar (CCA) according to Mwangi et al. (2006).

Pathogenicity tests

To determine if the isolates suspected to be Xcm were pathogenic to a susceptible banana cultivar, pathogenicity testing was done on three-month old tissue-cultured banana plantlets of cultivar Pisang Awak. The pathogen was introduced to the plant by injecting a 1 ml cell suspension (2 x 108 cells/ml) into the stem. The control was plantlets injected with a similar amount of sterile distilled water. The plants were kept under a shade net for three weeks. Disease symptoms were recorded every three days. In the second week, the first wilt was observed and re-isolation of the pathogen done as above. Bacterial colonies were sent to the Central Science Laboratory (CSL), United Kingdom (UK) for molecular characterization.

6 Mbaka et al Status of banana Xanthomonas wilt7 in Kenya-

RESULTS

The main banana cultivars grown in Western Kenya are shown in Figure 1.

Banana cultivars grown in Western Kenya

Cavedish Uganda green Apple banana Pisang Awak Boki boki Other EAHBs

Fig. 1 The major banana cultivars grown in Western Kenya (Source: Survey data 2006).

Of the major banana cultivars grown, Pisang Awak was the most common (in 44% of the farms. Banana production practices among the respondents are shown in Fig. 2.

Banana production practices in Western Kenya

100 90 80 Debudding 70 60 De-suckering 50 Use of own suckers 40 Suckers from neighbours 30 Tissue culture seedlings 20

Percentage of farmers of Percentage 10 0 1 Practices

Fig. 2: Banana Production practices in Western Kenya

(Source Survey data 2006).

Most of the respondents (90%) reported that they never removed the male bud. For Farm tools used in banana orchards were never disinfected. There was unregulated movement

7 Mbaka et al Status of banana Xanthomonas wilt8 in Kenya- of bananas from Uganda as observed at the Malaba border. Ninety five percent of the respodents reported that when establishing new orchards they got suckers from their fields or from neighbors. Tissue-cultured banana seedlings were planted by only five percent of the respondents. Most of the extension field officers and farmers could not differentiate Fusarium wilt from the symptoms of banana Xanthomonas wilt on the pictures. While the team was driving along a tarmac road connecting Malaba and Busia in

Chakol division of Teso district, a banana bunch with symptoms similar to those of BXW was spotted on one banana mat of Ndizi sukari (popularly known in Uganda as Kayinja) cultivar. The symptoms were: shriveled male bud, premature ripening of the fingers.

When cut across, the fingers had brownish colorations in the pulp. The disease was spotted in other fields in the division, Amukura division and Malakisi division of

Bungoma district. The banana cultivars affected were Bokiboki and Ndizi sukari. Yield losses were estimated at 70-90 %.

After 72 hours, bacteria colonies on YPGA and CCA were light yellow, circular, high convex, very mucoid which is characteristic of Xanthomonads. The inoculated plants showed a wilting and yellowing of leaves after three weeks. Seedlings inoculated with sterile distilled water had no wilt symptoms. The pathogen was re-isolated from the wilted plants and confirmed to be Xcm on the basis of growth characteristics. Fatty acid analysis done at the CSL, UK confirmed the pathogen to be Xcm.

DISCUSSION

The high incidence of BXW wilt in western Kenya could be explained by two factors: presence of the susceptible banana cultivars (apple banana, Boki boki and Pisang Awak),

8 Mbaka et al Status of banana Xanthomonas wilt9 in Kenya- and use of own banana suckers. Farmers in Western Kenya not only failed to remove the male bud but believed it can harm the banana. This pre-disposes the bananas to insect transmitted BXW infection. The bacteria can also be transmitted through contaminated tools yet farmers did not disinfect their tools. If the disease gets to areas like in Kisii region where pruning is practiced, one farmer can potentially spread disease to 200 plants per day through contaminated tools (Kubiriba and Tushemereirwe, 2006). Movement of banana material from the neighboring Uganda was unregulated. This could have brought in the initial Xcm inoculum. Failure of the field extension officers to correctly diagnose

BXW could have contributed to disease spread with the initial infection areas serving as sources of infection.

CONCLUSION

The survey established the presence of BXW in Western Kenya, an area currently producing 60 % of Kenya’s bananas. Most of the players in the banana value chain in the country are not aware of the disease and chances are that if no action is taken the disease will spread to other parts of the country. The disease poses a big threat to banana production in the country. This will lead to loss of income and food for the resource poor farmers. Business and employment in the public and private tissue culture laboratories in the country will also be lost.

RECOMMENDATIONS

Since BXW attacks all bananas and knows no boundaries, the starting point for disease management and/ or containment should be a country wide BXW situation analysis. This should be combined or followed by creation of awareness among all stakeholders.

Strategies developed and validated in Uganda should be validated in western Kenya and

9 Mbaka et al Status of banana Xanthomonas 10wilt in Kenya- adopted for disease management. These include destruction (cutting down, uprooting and burying) of infected plants to reduce inoculum in the fields (Kubiriba and

Tushemereirwe, 2006). Infected plants can also be destroyed by injecting them with herbicides like Glyphosate and 2 4 D and Paraquat (Blomme et al., 2006: Mwangi et al.,

2007). Insect transmission should be intercepted by removing the male bud (breaking with a forked stick) immediately the last flower opens (Kubiriba and Tushemereirwe,

2006). Cutting tools are disinfected in sodium hypochlorite (Jik®) and Savlon® (Mwangi et al., 2007) or by flaming.

ACKNOWLEDGEMENT

The authors would like to thank Catholic Relief services for financial support, Director

KARI and Centre Director KARI- Thika for logistical support, the Ministry of

Agriculture extension field officers for collaboration and farmers in Western Kenya for their cooperation during the survey.

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