Successful rescue and field establishment of native banana varieties severely affected by rhizome rot

Journal of Agriculture and Rural Development in the Tropics and SubtropicsVol. 113 No. 2 (2012) 147–154 ISSN: 1612-9830 – journal online: www.jarts.info Successful rescue and field establishment of native banana varieties severely aﬀected by rhizome rot Waman Ajit Arun †, a,∗, Pooja Bohra †, a, Konana Umesha b, Shivapur Channegowda Chandrashekar c, Bangalore Narayanappa Sathyanarayana a, Basawantanahalli Saddappa Sreeramu a aDepartment of Horticulture, University of Agricultural Sciences, GKVK Campus, Bengaluru, India bP.G. Centre, University of Horticultural Sciences (Bagalkot), GKVK Campus, Bengaluru, India cDepartment of Plant Pathology, University of Agricultural Sciences, GKVK Campus, Bengaluru, India Rhizome rot disease caused by Erwinia spp. is emerging as a major problem in banana nurseries and young plantationsworldwide. Management of the disease is possible only in the initial stages of development. Currently no method isavailable for rescuing plant material already infected with this pathogen. A total of 95 Nanjanagud Rasabale and 212Elakki Bale suckers were collected from diﬀerent growing regions of Karnataka, India. During nursery maintenanceof these lines, severe Erwinia infection was noticed. We present a method to rescue infected plants and establish themunder field conditions. Diﬀerences were noticed in infection severity amongst the varieties and their accessions. Fielddata revealed good establishment and growth of most rescued plants under field conditions. The discussed rescueprotocol coupled with good field management practices resulted in 89.19 and 82.59 percent field establishment ofpreviously infected var. Nanjanagud Rasabale and var. Elakki Bale plants, respectively.
Keywords: Ney Poovan, rescue technique, Silk banana, soft rot, tip over Bananas and plantains, staple food crops in many tropical countries, are aﬀected by a variety of pests and diseases which can cause catastrophic yield losses INIBAP: International Network for the Improvement of Ba- (Jones, 2000). Such biotic and abiotic challenges have nana and Plantain, been the major reasons for initiating crop improvement length of youngest open leaf, programmes worldwide (Ploetz, 2004), including the es- Nanjanagud Rasabale, tablishment of International Network for the Improve- width of youngest open leaf ment of Banana and Plantain (INIBAP) in France (Bud-denhagen, 1993). Apart from Fusarium wilt, Sigatokaleaf spot, banana bunchy top and other viruses (Sathi- ∗ Corresponding author amoorthy, 1994), a large number of other diseases have started negatively impacting banana production (Ploetz, Phone: +91 9986 430392Department of Horticulture, University of Agricultural Sciences, 2004). Rot of the rhizome and pseudostem caused by GKVK campus, Bangaluru 560065, India Erwinia spp. were considered to be of little significance † Equal contribution to the present work in the past (Thwaites et al., 2000), but their severity has Waman Ajit Arun et al. / J. Agr. Rural Develop. Trop. Subtrop. 113 - 2 (2012) 147–154 increased in recent years in many growing countries in- istics surveys were undertaken in growing districts of cluding India (Manoranjitham et al., 2010; Singh et al., Karnataka and suckers of putative natural variants were 2010; Thammaiah et al., 2010; Patel et al., 2011).
Rhizome Rot, also known as Soft Rot or Tip Over Elakki Bale syn. Safed Velchi (Ney Poovan group, Disease is commonly observed during the first 3–4 Musa AB), an elite native variety of banana, is popular months after planting under high temperature condi- amongst farmers in the states of Karnataka and Maha- tions, especially during late summer and rainy season. It rashtra, India, as it fetches almost double the price of is caused by the bacteria Erwinia carotovora (Syn. Pec- commercially dominant Cavendish bananas in the do- tobacterium carotovora) and E. chrysanthemi, which mestic market owing to its superior quality fruits with are opportunistic residents of banana soils (Thangavelu, sweet pulp. This otherwise elite variety suﬀers from the 2009). They enter the plant through wounds and spread problem of lodging especially in the high wind areas due across fields through water and infected planting ma- to taller stature of the plants. To assess the natural diver- terial (Ravishankar, 2010). As suggested by the name sity for the plant height and other agronomic character- ‘soft rot', the plants infected with the pathogen show istics, surveys were done in ten districts of Karnataka typical symptoms of rotting in the rhizome, yellowing and desired clones of this variety were collected.
and subsequent drying of leaves, and breaking of pseu- During the period between collection and field plant- dostem at the collar, hence the term ‘tip over disease'.
ing, the suckers were planted in pots containing sand Symptoms of this disease were first reported from and were maintained in the polyhouse.
Honduras in 1949, and its widespread distribution cessions were from diverse fields (mainly from poorly is made evident by reports from diﬀerent regions maintained orchards as main objective was screening for and countries: Israel (Volcani & Zutra, 1967), Cen- Fusarium resistant lines) even after appropriate main- tral America (Wardlaw, 1972), Jamaica (Shillingford, tenance, the plants got severely infected with rhizome 1974), Papua New Guinea (Tomlinson et al., 1987) and Though diﬃcult, the disease is curable in the India (Singh, 1990). Incidents of Rhizome Rot have early stages of infection, if proper control measures are become alarming in India in the recent past and for adopted. However, the survival of the plant becomes this reason it is the only bacterial disease against which less likely if the infection is severe and no technique is breeding objectives for crop improvement in banana currently available to rescue the plants which are heavily were set (Sathiamoorthy, 1994). The occurrence of Rhi- infected with this pathogen. Eﬀorts were made to save zome Rot has been reported from the states of Kerala the suckers from the pathogen and this paper presents (Singh, 1990), Gujarat (Singh, 1990; Patel et al., 2011), the method used for rescuing heavily infected banana Tamil Nadu (Manoranjitham et al., 2010), Bihar (Singh suckers and establishing them successfully in the field.
et al., 2010) and Karnataka (Thammaiah et al., 2010).
We hope this report will be of use to researchers dealing Apart from aﬀecting banana fields, soft rot has been with the same pathogen in their germplasm nurseries, reported as a major limiting factor in weaning of mi- so that they may avoid losing their valuable germplasm cropropagated plantlets (Thangavelu, 2009). The ex- and to farmers who commonly face this problem during tent of deaths due to soft rot in secondary hardening has the initial banana field establishment phase. Similarly, been reported to be around 2–5 % (Thomas et al., 2011).
it may be a help to nurserymen, who deal with such Furthermore, E. carotovora is known to contaminate in infections when attempting to multiply banana through vitro cultures (Leifert et al., 1994), causing big losses to commercial tissue culture units.
Nanjanagud Rasabale (Musa AAB, Silk banana) is Materials and methods
the choicest variety of the state of Karnataka, India. It isan ecotype which has been given the status of Geograph- The present investigation was carried out in a natu- ical Indication Crop by the Government of India owing rally ventilated polyhouse and the experimental fields of to its appealing aroma, attractive bright yellow colour the Department of Horticulture, University of Agricul- and delicious pulp (http://ipindia.nic.in/girindia/). How- tural Sciences, Bengaluru, India. The experimental field ever, this ecotype is highly susceptible to Panama Dis- is located in the eastern dry zone of Karnataka (12°58' ease or Fusarium Wilt caused by the fungus Fusarium N; 77°35' E) at an altitude of 930 meters above mean oxysporum f.sp. cubense and as a result its production area has been drastically reduced from about 500 ha in1960 to but a few hectares presently (Venkatachalam Germplasm collection and nursery management et al., 2006). In order to assess the diversity for Panama Exploration was carried out from January 25 th, 2011 disease resistance, yield and other agronomic character- to March 25th, 2011 in eleven districts of the state of Waman Ajit Arun et al. / J. Agr. Rural Develop. Trop. Subtrop. 113 - 2 (2012) 147–154 Karnataka. In case of Nanjanagud Rasabale (NR), ex- of 2 × 2 m and filled with one kg vermicompost, 15 kg tensive field surveys were carried out in the Mysore and farm yard manure and top soil. As a precautionary mea- Chamarajanagar districts of the state. Clones showing sure, holes were sterilized with one litre 0.2 % MEMC various degree of resistance to Panama Disease were solution, one day prior to transplanting. To provide pro- selected and the maximum possible numbers of suck- tection against nematodes, 50 g of Tatafuran (carbofu- ers were procured from the selected plants. For collec- ran 3G) was added to and mixed into the soil in the tion of Elakki Bale (EB), dwarf stature, resistance to planting holes before planting. Plants were irrigated Panama Disease and good agronomic parameters were just after planting and subsequent operations were per- the selection criteria and collection was done in ten formed following the package of practices developed by districts: Bengaluru (Urban), Bengaluru (Rural), Chi- the National Research Centre for Banana, Trichy, India tradurga, Davanagere, Hassan, Kolar, Mandya, Mysore, Ramanagara and Tumkur.
Collected suckers were brought to the polyhouse and Recording observations and statistical analyses washed with water to remove the adhering soil parti-cles. Dried leaf sheaths and roots of the suckers were Survival and field establishment of rescued plants was removed using a sharp knife prior to soaking in 0.2 % calculated based on the number of plantlets that sur- Bavistin solution for 20 minutes. Treated suckers were vived the first four months after transplantation. No then planted in polypropylene pots containing steril- Erwinia disease was observed in the field thereafter.
ized sand. Pots were maintained in polyhouse before Parameters such as plant height (cm), collar diameter planting in the field. After one month, a plant was ob- (mm), number of suckers, number of leaves, length served to have bacterial rot symptoms which were con- (cm) (LYOL) and width (cm) (WYOL) of youngest open firmed after its examination and within the period of one leaf were recorded at two months intervals: two and week, the number of infected plants increased. Soon af- four months of planting, as this is the most crucial pe- ter the incidence was noticed, the plants were drenched riod in plantlet establishment. Infection resulted in un- with a solution containing copper oxychloride (0.3 %, equal numbers of replications in most of the accessions; Blitox, Tata Rallis, Mumbai, India) plus 600 mg L−1 hence the mean of the available plants was taken into Krocin AG (streptomycin sulfate 9 % plus tetracycline consideration for comparing the growth performances.
hydrochloride 1 %, Krishi Rasayan, India) and the treat- Paired t-test was done using the Web Agri Stat Pack- ment was repeated at weekly intervals for three weeks, age (WASP, v. 2.0, ICAR Research Complex for Goa) as the spread increased in intensity resulting in the loss to compare the growth parameters between establish- of many accessions.
ment (two months after planting) and post establishmentphase (four months after planting).
Rescue method As the severity of the disease was high and initial control measures were not eﬀective, plants were giventreatments of methoxy ethyl mercuric chloride (MEMC,Bagalol, United Phosphorus Limited, India). For treat- Development of disease in nursery ment, the infected tissue was scooped out until healthy Suckers of all accessions in both varieties started white rhizome tissue was revealed. Care was taken that sprouting 10–15 days after planting in pots (Fig. 1a no rotten portion remained on the rhizome surface. Such and 1b) and good growth was noted within one month shaved rhizomes were treated with 0.2 % MEMC solu- (Fig. 1c and 1d). The symptoms of rhizome rot started tion for 5 minutes and were planted in new pots con- about four weeks after planting in pots with yellowish taining a sterilized mixture of red soil and coir pith in brown blighting from the leaf margins, progressing to- equal proportions. The suckers had little or no roots and wards the midrib. The aﬀected portion started drying the size was reduced to a piece of about 2.5 cm thick- up and almost all mature leaves showed similar symp- ness of healthy tissue. Plants were watered judiciously toms. During this initial phase, the rhizome appeared to as and when required. To promote root formation and have a hard surface but showed typical dark brown rot- plant growth, plants were sprayed twice with mono am- ting when excavated from sand and cut crosswise (Fig.
monium phosphate (0.2 %) at weekly interval.
1e). As the disease progressed, the rotting extended tothe whole rhizome except for a bit of healthy tissue near Field planting and aftercare base of the plant. The rotten portion looked like deli- After 45 days of treatment, plants were transplanted cate black threads and was moist (Fig. 1f). During this to the field. Field was prepared by ploughing twice and phase, the pseudostems broke at the collar region and levelling. Pits of 60×60×60 cm were dug at the spacing were unrecoverable.

Waman Ajit Arun et al. / J. Agr. Rural Develop. Trop. Subtrop. 113 - 2 (2012) 147–154 Fig. 1: Development of disease in nursery: (a & b) sprouting of NR and EB suckers, respectively; (c & d) growth of NR and EB
suckers; (e) rotting of the rhizome due to Erwinia sp.; (f) advance stage of disease development.
Waman Ajit Arun et al. / J. Agr. Rural Develop. Trop. Subtrop. 113 - 2 (2012) 147–154 Eﬃciency of rescue methodology increased significantly, whereas number of functional Data on the number of plants that died due to Rhi- leaves on the plant and the width of youngest opened zome Rot at each stage is presented in Table 1. Mor- leaf remained non-significant (Table 2). Seven acces- tality was divided into post infection loss due to Er- sions (NR 8, NR 12, NR 14, NR 18, NR 19, NR 21 winia in polyhouse and post treatment (rescue) loss un- and NR 24) showed excellent establishment and growth der field conditions. Diﬀerent accessions of NR showed during post establishment phase.
various degrees of susceptibility to rhizome rot. Out In case of EB, highly significant diﬀerences were of a total of 95 suckers from 27 diﬀerent accessions, recorded for parameters such as plant height and col- 22.11 % (21 suckers) died in the nursery, wiping out lar diameter, number of leaves, and length and width all the replications of accession NR 1. About 14 acces- of youngest opened leaf. Two to four fold increases sions showed tolerance to rhizome rot: no deaths were in plant height was observed amongst the accessions reported from these lines, even after infection. Thir- between establishment and post- establishment phases teen of these lines maintained tolerance and recovered (Table 3). However, apparent variability was observed on transplanting in the field with 100 % establishment.
among the accessions. Collar diameter also followed the The overall post treatment survival rate under field con- same trend and significantly improved in two months.
ditions was 89.11 %.
There were no significant diﬀerences in number of suck-ers, though. The reason might be the smaller size of the Table 1: Sucker losses due to rhizome rot under diﬀerent con-
chopped rhizome in which only a few lateral buds re- ditions and field establishment of the rescued plants. mained. Many accessions did not produce any suckerswithin the first four months after planting. This might be due to genetic diﬀerences between them. Cumulative count of leaves improved in a highly significant manner where the range of functional leaves increased from 6– No. of suckers at collection 10.7 to 8–14 leaves/plant. Length of youngest opened Suckers lost in nursery due to Erwinia leaf also increased over time, which might be due to in- creasing petiole length of the succeeding leaves. Due No. of suckers planted in the field to normal root growth, plant metabolism also improvedand hence, leaf length as well as width increased in this No. of plants in field 120 DAP formative phase of plant growth.
Field establishment (%) The losses of EB in nursery stage was 59.43 %, result- ing in the complete loss of thirty one accessions. Suck- The name soft rot derived from the characteristic soft ers of eleven accessions, though infected, survived the decay of the fleshy tissues of the rhizome of infected ba- nursery stage. However, one of these lines was killed nanas. Banana, like most tuberous and bulbous plants, oﬀ by the pathogen under field conditions, probably due has higher carbohydrates in the rhizome tissues and to the systemic presence of the pathogen and the shock these storage organ tissues have cells in semi dormant of transplantation. The rhizome rot survival rate of EB condition. When such tissues get infected with Erwinia, was 82.59 %.
they become soft, watery or slimy in consistency andwith the progress of the rot, water exudes from the af- Post-rescue field establishment fected region (Walker, 2004). These symptoms were In order to study the eﬀect of the rescue protocol on also noticed during the course of present investigation.
subsequent establishment of plants in the field, observa- The softening is mainly caused by the action of pectic tions were recorded on diﬀerent growth parameters two enzymes on the storage tissues (Mehrotra & Aggarwal, and four months after planting. Data is presented in Ta- bles 2 and 3. Main objective was to find out the growth Literature review suggests that this disease has not diﬀerences between establishment (Fig. 2) and post es- been recorded in Ney Poovan nor in Nanjanagud Rasa- tablishment phases (Fig. 3). The comparison of these bale, though rhizome rot has previously been reported in phases using paired t - test in NR revealed highly sig- diﬀerent banana cultivars: Gros Michel (Stover, 1959), nificant diﬀerences for parameters such as plant height Nendran (Singh, 1990), Dwarf Cavendish (Singh, 1990; and collar diameter. The number of suckers were also Patel & Shukla, 2010) and Grande Naine (Patel & found to be significantly diﬀerent, but were observed Shukla, 2010) from diﬀerent banana growing regions.
to be less in post establishment phase than in the es- After conducting studies on bacterial corm and rhi- tablishment phase. Length of the youngest opened leaf zome rot in ten diﬀerent banana varieties in Papua New

Waman Ajit Arun et al. / J. Agr. Rural Develop. Trop. Subtrop. 113 - 2 (2012) 147–154 Table 2: Growth performance of the rescued accessions of NR during establishment (60 days after planting, DAP) and post-
establishment (120 DAP) phases.
Plant Height (cm) Collar diameter (mm) Number of suckers Number of leaves ns: non-significant, *: significant at 5 % and **: at 1 % level of significance using paired t test Table 3: Growth performance of the rescued accessions of EB during establishment (60 DAP) and post-establishment (120 DAP)
phases.
Plant Height (cm) Collar diameter (mm) Number of suckers Number of leaves ns: non-significant, *: significant at 5 % and **: at 1 % level of significance using paired t test Fig. 2: View of a recently planted field of rescued suckers (a); close up view of a newly planted sucker (b).
Fig. 3: View of a field of successfully established NR (a) and EB (b) plantlets.
Waman Ajit Arun et al. / J. Agr. Rural Develop. Trop. Subtrop. 113 - 2 (2012) 147–154 Guinea, Tomlinson et al. (1987) suggested that the ge- netic makeup of a plant plays a vital role in deciding itssusceptibility to a pathogen. Varied degrees of incidence Erwinia Rot, though considered as a minor disease, and disease severity have been recorded in diﬀerent vari- can cause considerable damage if not managed prop- eties in previously described reports e.g. 37 % in Dwarf Diversity existed in the accessions as diﬀer- Cavendish, 20-30 % in Grande Naine and 10–15 % in ent lines showed varied levels of infection. The res- Gros Michel. The variable resistance reactions exhib- cue method described was highly eﬀective for rescuing ited by diﬀerent varieties and accessions in the present suckers heavily infected with the pathogen. A proper study may therefore be due to genetic diﬀerences be- package of rescue techniques and appropriate supply of tween the plants.
water, nutrients and growth conditions could result in Better vegetative growth improves the reproductive improved establishment and subsequent growth of res- growth in all the plants, and banana is no exception.
cued plants.
Positive correlation between yield parameters and vege- tative growth parameters, mainly plant height and girth Waman Ajit Arun and Pooja Bohra are thankful to the has been reported in Yangambi km 5 and Datil culti- Department of Science and Technology, Government of vars in Costa Rica (Vargas & Sandoval, 2005). Also, India for providing the financial assistance in the form of the plant height and collar diameter were found to be INSPIRE fellowship. Thanks are also due to the farm- highly correlated with the days for flowering in trials in ers and oﬃcials of the State Department of Horticulture, Nigeria (Blomme et al., 2006).
Government of Karnataka for their help during survey The plants which get properly established in the field and collection.
can absorb nutrients supplied to them eﬀectively andthus can carry out cell division more eﬃciently. Dur-ing transplanting, most of the plants had a few or no roots and thus it was diﬃcult for the plants to absorbany nutrients from the soil. As opined by Blomme et al. Blomme, G., Swennen, R., Ortiz, R. & Tenkouano, A.
(2006), if optimum growing condition along with proper Root system and shoot growth of banana supply of water, nutrients and sunlight is provided, ba- (Musa spp.) in two agro-ecological zones in Nigeria.
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