PLANTAIN BREEDING AT CRBP : STRATEGIES, RESULTS AND OUTLOOK

Kodjo Tomekpé, Erik Auboiron, Pascal Noupadja and Eric Fouré

Centre de Recherches Régionales Sur Bananiers et Plantains (CRBP)
B.P. 832, Njombé
Cameroon.

Keywords: Musa ssp., Mycosphaerella fijiensis, black sigatoka, embryo rescue, hybrid.

Abstract

Plantain breeding started in 1992 at the CRBP aiming to develop black sigatoka resistant
plantain tetraploid hybrids by crossing plantain (AAB) with black sigatoka resistant
diploid bananas (AA). Calcutta 4 and M53 are the most popular diploid parents. Calcutta
4 is non-parthenocarpic and highly resistant to black sigatoka. M53 is a parthenocarpic
hybrid, which was bred by Musa breeders in Jamaica, and shows partial resistant to black
sigatoka. Forty-six triploid plantain cultivars set seed (0 to 183 seeds per bunch) after
hybridization with pollen of diploid bananas. The seed set depends on cultivars, period of
pollination and pollen source. Three-hundred hybrids rescued by embryo culture are
undergoing preliminary evaluation under high pressure of black sigatoka. These hybrids
show distinct ploidy levels: diploids, tetraploids, triploids and heptaploids. About 30% of
hybrids were aneuploids. Eight black sigatoka resistant tetraploid hybrids have already
been selected through two cycles of preliminary evaluation. These hybrids are generally
shorter and more vigorous than their female plantain parents.They exhibit a strong
ratooning and early flowering but their fruit filling time is longer, and produce larger
bunch than plantain in high pressure of black sigatoka. Unlike plantain cultivars, these
hybrids are male- and female-fertile. Four of them are currently under clonal evaluation.

1. Introduction

Plantain (Musa spp., AAB) is a staple food for about 70 million people in the
humid forest of West and Central Africa. It is mainly produced by smallholders and
provides an important source of rural income. The plantain exhibits a wide spectrum of
phenotypic variation as a result of the accumulation of somatic mutations, particularly in
Central and West Africa. This region is therefore considered a secondary center of
diversification. All cultivars are susceptible to black sigatoka (Mycosphaerella fijiensis),
nematodes and weevil borer, but tolerant to race 1 of Panama disease (Fusarium
oxysporum f.sp. cubense). Most clones are tall and very vulnerable to wind. Nematode
damages increase their vulnerability to wind. Since smallholders cannot benefit from
expensive agrochemical control of these constraints and due to ecological limits of such
control, development of resistant cultivars by breeding seems to be an interesting option.
Plantain breeding in Africa received increased attention only about 10 years ago
when black sigatoka spread rapidly into Central and West Africa (Fouré et al., 1984;
Wilson and Buddenhagen, 1986; Swennen and Vuylsteke, 1988, 1989). This disease is
caused by an airbone fungus Mycosphaerella fijiensis Morelet and reduces plantain yield
by 30 to 50 %. Current breeding approaches are based on the traditional scheme of 3x x
2x crosses to produce tetraploids. In this scheme, plantain cultivars are crossed with AA
black sigatoka resistant diploids in order to obtain resistant tetraploids. Many resistant
hybrids were produced through this breeding method (Vuylsteke et al., 1993a, 1993b;
Rowe and Rosales, 1993; Bakry and Horry, 1992). Almost all the hybrids selected from
African plantains were derived from two plantain cultivars and the non-parthenocarpic
wild banana Calcutta 4. Since a large plantain collection is available at CRBP, a breeding
program was initiated in 1992 using a wide range of plantain cultivars plus wild and
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improved AA male parents (Jenny et al.,1994). Hybdrization between distinct female-
fertile plantains and diverse male parents will enhance the variation among the hybrids,
thereby increasing the chance for selection of resistant and adapted hybrids for
smallholders. This article reports results obtained by this breeding approach at CRBP.

2. Materials and methods

2.1. Selection of male parents

Plantain breeding at CRBP depends on M53, a synthetic diploid hybrid developed

by Musa breeders in Jamaica. This diploid hybrid has good agronomic characteristics
(e.g. large bunch, long parthenocarpic fruits and strong ratooning) with pollen fertility and
partial resistance to black sigatoka. Due to its high level of heterozygosity, M53 is
undergoing inbreeding selection to develop improved lines for plantain breeding. A few
individuals from selfing M53 have already been selected and crossed with fertile
plantains. Their progenies have now been planted for preliminary evaluation.
Another M. acuminata banana intensively used as male parent is Calcutta 4. This
wild banana is highly male and female-fertile, highly resistant to black and yellow
sigatoka, and tolerant to the burrowing nematode Radopholus similis. This diploid
accessions shows a strong ratooning but exhibits poor agronomic characteristics such as
small and non parthenocarpic fruits. M. acuminata banksii Banksii Type has been also
tested as male parent. This diploid accession exhibits a great number of hands and high
resistance to black sigatoka. It is generally considered that the M. acuminata genomes of
plantains may originate from the subspecies banksii.

2.2. Pollination, seed collection, embryo rescue and production of hybrid plants

Sixty-nine plantain cultivars including 63 French plus 6 False Horn and French
Horn plantains were hand-pollinated in the morning (7 am to 10 am). Most cultivars were
pollinated one to two times while eight fertile clones were intensively hybridized.
Bunches were harvested at maturity and seeds were collected. Embryos from viable seeds
were extracted after seed sterilization and in vitro rescue techniques (Jenny et al., 1994).
After transfer of rooted plantlets in the nursery and in pots, hybrids were planted in the
field for preliminary evaluation under high pressure of black sigatoka.

2.3. Determination of black sigatoka response

Host responses to black sigatoka were recorded in the field according to the
scoring method developed by Fouré (Fouré et al., 1984, 1990). In the preliminary
assessment, the youngest leaf spotted (YLS) from the first open leaf until flowering was
recorded in one or two plants per hybrid. Plant host responses were considered as
susceptible (low YLS), partially resistant (increasing YLS) and highly resistant.

3. Results and discusion

3.1. Female fertility of plantains

Among the 69 plantain cultivars screened for female fertility, 46 set seeds after
hand-pollination. The environment at Nyombé (southwest Cameroon) appears to be
favorable for cross-breeding a wide range of plantains. Only one False Horn plantain
produced seeds, while the others were French plantains, which produced up to 183 seeds
per bunch. Seed set depended on clone, period of pollination and pollen source (Table 1).
French Clair and French Sombre showed the highest fertility. The 183 seeds were
produced after pollinating French Clair with M53. The fertile False Horn Plantain N°17
was hybridized two times and produced 51 seeds and nine hybrids of which four were
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aneuploids, four diploids and one triploid. Eight giant and medium-sized cultivars
selected for their capability to produce interesting hybrids are being hybridized.

3.2. Male fertility

Calcutta 4 was generally more fertile than M53 except with French Clair that was
particularly successfull to produce seeds and hybrids when pollinated with M53 (Table 1).
Except M53H50-048 and M53H50-50, the male parents derived from M53 selfing
exhibited low fertility. A few hybrids, including tetraploids, were obtained from these
selfed progenies. M. acuminata banksii Banksii Type showed very low fertility.

3.3. Ploidy level of plantain hybrids

The ploidy level was determined by flow cytometry. Plantain progenies were
divided into 70% euploids and 30% aneuploids. Among the euploids, diploids and
tetraploids were 50% and 40%, respectively, whereas triploids and heptaploids accounted
for 10%. French Clair seems to be the most efficient cultivar for tetraploid hybrids
production. The high proportion of tetraploid hybrids shows that gamete restitution is
relatively frequent in plantain and supports early results obtained at the International
Institute of Tropical Agriculture in Nigeria (Vuylsteke et al., 1993).

3.4. Preliminary description of tetraploid plantain hybrids

3.4.1. Resistance to black sigatoka

Preliminary evaluation of black sigatoka response showed that almost all the
tetraploid hybrids are resistant to black sigatoka. The tetraploid hybrids derived from the
highly resistant Calcutta 4 exhibited partial resistance to black sigatoka confirming the
report of Vuylsteke et al. (1993). None of these hybrids exhibited the high resistance of
Calcutta 4, which indicates a complex genetic control ensuing from interactions involved
in the expression of high resistance to black sigatoka.
Except a few highly resistant and susceptible hybrids, all the tetraploid hybrids
from M53 are partially resistant to black sigatoka. The occurrence of highly resistants
hybrids derived from the partially resistant M53 was not surprising. This hybrid is highly
heterozygote and has in its pedigree the highly resistant M. acuminata ssp. banksii
accessions Samoa Type and Paka. In addition, 60% highly resistant and 40% partial
resistant selfed progenies from M53 were observed, which suggest that M53 has genes for
both partial and high resistance to black sigatoka.

3.4.2. Agronomic performance

Among the 150 preliminary evaluated in the field during two cycles, eight black
sigatoka resistant tetraploid hybrids were selected for their improved agronomic
characteristics. These hybrids were derived from medium and giant-sized plantain
cultivars as females and M53 and Calcutta 4 as male parents. Four of these tetraploid
hybrids are undergoing clonal evaluation under high pressure of black sigatoka. Table 2
list the agronomic characteristicsc of these selected hybrids through two cycles of
preliminary assessment. Ten other tetraploid hybrids proved to be promising after
preliminary observations on the first cycle.
The tetraploid hybrids look very similar to the plantains and exhibit long
parthenocarpic fruits. Moreover, most of them show short plant height, improved
suckering and early flowering. Preliminary data indicate that these hybrids have higher
bunch weight than their fungicide-treated plantain parents. Most of the tetraploid hybrids
derived from pollination with Calcutta 4 do not exhibit its undesirable agronomic traits
such as non-parthenocarpic, small fruits. This result confirms the observations reported by
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Vuylsteke et al. (1993a). However, these hybrids exhibit some weaknesses such as long
fruit filling time, and drooping leaves. Unlike tetraploid hybrids from Calcutta 4, the
leaves of progenies derived from M53 are not drooping. The most important defect of
these hybrids as new cultivars is their high male- and female-fertility that allow the
occurrence of seeded fruits. A preliminary evaluation of the physico-chemical characters
shows that dry matter content of most hybrids is lower than in their plantain parents.
Moreover, eating quality of some hybrids needs to be improved.

3.5. Description of plantain-derived diploid

Plantain-derived diploid hybrids have also been recovered from 3x x 2x crosses. A

high percentage of these diploid hybrids are susceptible to black sigatoka, especially those
derived from M53. The proportion of interesting diploid hybrids was low and they were
mostly derived from M53. These plantain-derived diploid hybrid exhibit a large bunch
and fruit characteristics of plantains (Tomekpé et al., 1995). Almost all the diploid
hybrids from Calcutta 4 look very similar to their male parent and exhibit poor agronomic
characteristics. The difference between diploid and tetraploid hybrids would be partly due
to gene dosage effects (Ortiz, 1995; Tomekpé et al., 1995). The promising plantain-
derived hybrids with resistance to black sigatoka, big bunch and long fruits, would
provide useful genetic variation for plantain improvement.

4. Outlook of plantain breeding at CRBP

CRBP aims to obtain black sigatoka resistant tetraploid hybrids that keep the fruit
characteristics of the triploid plantain parent. The results reported in this article clearly
demonstrate that resistant improved hybrids can be developed in a relatively short period.
The variation observed in the tetraploid hybrids and the selection of plantain-derived
diploid are very promising for further development of new plantain cultivars at CRBP.
The high number of female-fertile plantains makes CRBP an ideal center for plantain
breeding. The improved hybrid M53 has proved to be an efficient male parent for plantain
breeding. Plantains pollinated by M53 produced more viable embryos and promising
tetraploid hybrids with no breakage of leaf petiole. M53 is also an efficient parent to
obtain useful plantain-derived diploid hybrids.
Four promising hybrids selected from a preliminary trial are being clonally tested
to determine their black sigatoka resistance stability and agronomic performance.
Assessment of physico-chemical traits and culinary quality will be undertaken soon and
will be followed by multilocational testing in Cameroon and in other Central and West
African locations to assess adaptability and genotype-by-environment interactions.
Nematode resistance will be among the new breeding targets. Moreover, new
tetraploid hybrids will be obtained to improve dwarfism. ). Priority will be given to a new
breeding strategy at CRBP in which resistant plantain triploid hybrids are developed by
4x x 2x (⇒3x) genotypes. The diploid parents will be wild BB accessions or improved
AA genotypes as per the genome of the tetraploid parents.This approach was developed
by CIRAD-FLHOR in Guadeloupe with very promising results (Horry et al., 1993). To
obtain triploid hybrids with most plantain characteristics, selected plantain-derived
diploid hybrids will be used in this scheme. The main advantage of this long-term
approach is the production of hybrids similar to the available triploid cultivars but with
disease and pest resistances plus potential male sterility, thereby avoiding seeded fruits.
Non-conventional breeding initiated at CRBP in collaboration with CIRAD-
FLHOR and CATIE focuses on two methods : the development of somatic hybridation by
protoplast fusion between AA cooking diploids and B haploids, and genetic
transformation by biolistic. For both purposes, somatic embryogenesis and cell
suspensions from male flowers (Escalant et al., 1993) have already been established in the
tissue culture laboratory of CRBP. Somatic embryogenesis cultures and cell suspensions
will the master-key for the successful application of these methods.
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Acknowledgements

The authors thank the 7th FED and STD3 programs of the European Economic
Community for their financial support. We are grateful to P. Ollitrault for assisting us in
flow cytometry analysis and to J. Tchango Tchango for analysis of dry matter content.

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