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219© Springer International Publishing AG 2017
H. Campos, P.D.S. Caligari, Genetic Improvement of Tropical Crops,
DOI10.1007/978-3-319-59819-2_7
Chapter 7
Bananas andPlantains (Musa spp.)
AllanBrown, RobooniTumuhimbise, DelphineAmah, BrigitteUwimana,
MosesNyine, HassanMduma, DavidTalengera, DeborahKaramura,
JeromeKuriba, andRonySwennen
7.1 Introduction
With a production of 145 million metric tons worldwide (worth 26.5 billion Euro),
banana (Musa spp.) is one of the world’s most important staple food crops and argu-
ably the world’s most popular fruit in terms of international trade (FAO 2014).
Banana and plantains (Musa spp.), collectively referred to here as bananas, are
grown in more than 135 countries and found in most tropical and subtropical regions
around the world. While industrialized nations view banana primarily as a dessert
A. Brown (*) • H. Mduma • R. Swennen
International Institute of Tropical Agriculture, Arusha, Tanzania
e-mail: a.brown@cgiar.org; h.mduma@cgiar.org; r.swennen@cgiar.org
R. Tumuhimbise • J. Kuriba
National Agricultural Research Organization, Kampala, Uganda
e-mail: rtumuhimbise@hotmail.com; jkubiriba2012@gmail.com
D. Amah
International Institute of Tropical Agriculture, Ibadan, Nigeria
e-mail: D.Amah@cgiar.org
B. Uwimana • M. Nyine
International Institute of Tropical Agriculture, Kampala, Uganda
e-mail: B.Uwimana@cgiar.org; m.nyine@cgiar.org
D. Talengera
National Agricultural Research Laboratories, Kampala, Uganda
e-mail: dtalengera@yahoo.com
D. Karamura
Bioversity International, Kampala, Uganda
e-mail: d.karamura@cgiar.org
220
item, many regions of the developing world consider cooking bananas and plantains
as essential staples that contribute signicantly to the caloric intake of low-income
subsistence farmers. Although sensitivity to photoperiod has been noted in certain
cultivars (Fortescue etal. 2011), banana is an almost nonseasonal crop that reliably
provides a carbohydrate source year-round which makes it vitally important to both
nutrition and food security. Propagation by farmers is commonly through suckers or
side shoots originating from lateral buds at the base of the main plant. Multiple
fungal and bacterial pathogens present serious constraints to production of bananas,
as does the occurrence of insects and nematodes (Jones 1999). Viral diseases caused
by banana streak virus (BSV), cucumber mosaic virus (CMV), banana bract mosaic
virus (BBMV), and the emerging banana bunchy top virus (BBTV) are also receiv-
ing increased attention (Kumar etal. 2015). The predominance of these biotic agents
differs from region to region, but most are found throughout the banana production
regions in Asia, Africa, and the Americas and represent common targets for plant
improvement worldwide. As with all crops, abiotic factors associated with climate
change such as drought and heat stress also present considerable challenges to pro-
duction (van Asten etal. 2011; Wairegi etal. 2010), but arguably the single greatest
constraint to genetic improvement is the narrow genetic basis of most cultivated
bananas (Hippolyte etal. 2012) and the physiological and reproductive barriers of
the plant itself (Ssesuliba etal. 2008; Fortescue and Turner 2004; Dumpe and Ortiz
1996). Reproductive barriers limit sexual recombination in banana and hinder plant
improvement. While all of the seed-bearing progenitors of modern banana cultivars
are diploid in nature, those that have been cultivated for consumption are primarily
seedless triploids. Female fertility of triploids has been described, but seed set is
generally extremely limited which complicates breeding efforts and intensives
resources and time required to develop superior varieties with enhanced resistance
to multiple biotic agents and abiotic agents. Banana improvement is further compli-
cated by parthenocarpy, reduced male fertility in some cultivars, low seed viability,
irregular meiotic behavior, long generation times, and diverse genomic congura-
tions (Ortiz 2013, 2015; Ortiz and Swennen 2014). To date, the limited progress that
has been achieved in banana breeding has occurred through crossbreeding
approaches that involve hybridization followed by phenotypic selection among half
sibs and/or full sib progenies.
7.2 Banana Classication
Banana is a monocotyledon herbaceous plant represented by three genera (Musa,
Ensete, and Musella) within the family Musaceae of the order Zingiberales (De
Langhe etal. 2009). The genus Ensete consists of monocarpic, unbranched herba-
ceous plants that rarely produce suckers and are used for food, ber, and ornamental
purposes. They resemble banana, but their oversized, edible corms and wide-
spreading and immensely long, paddle-shaped leaves with crimson midribs make
them very distinctive. Their fruits are similar in appearance to those of banana, but
A. Brown et al.
221
are dry, seedy, and inedible (Deckers etal. 2001). The most recognizable member of
this genus is perhaps the false or Abyssinian banana (E. ventricosum) that plays a
signicant role in Ethiopian agriculture and food security (Tsegaye and Struik
2002).
The genus Musa consists of cultivated triploid cultivars and clones propagated
through vegetative methods with limited genetic variation beyond what could be
expected through somaclonal variation (and perhaps epigenetics) and the diploid
wild progenitors of these cultivars that are capable of sexual recombination. More
than 60 species within four recognized sections of the genus Musa have been
described, but the taxonomy of Musa and the relationship between wild and culti-
vated bananas are far from settled (De Langhe et al. 2009; Janssens etal. 2016).
Almost all diploid species are native to Southeast Asia, from India and Thailand to
New Guinea and Queensland, Australia (Simmonds 1987). Edible bananas, with the
exception of the Fe’i group of the Australimusa section, are derived almost exclu-
sively from two species, Musa acuminata and Musa balbisiana of the section Musa
(Dodds 1945). M. acuminata and M. balbisiana are diploid (2n = 2x = 22) in their
base genomic complements and designated as AA and BB, respectively (Simmonds
and Shepherd 1955). In addition to monospecic cultivars (AA, BB), interspecic
diploid clones (AB) are also recognized. Higher-order combinations of the AA and
BB base genomes arose through chromosome restitution at meiosis, to produce dis-
tinct groupings at the triploid (AAA, AAB, ABB groups) and occasionally tetraploid
levels (Simmonds 1962). Triploids, due to their optimal vigor and seedless charac-
teristic, are the preferred conguration for most consumers throughout the world
(Simmonds 1987). The rare tetraploid cultivars tend to be physically larger but have
relatively small bunches, while most diploid cultivars tend to be weaker plants with
smaller bunches (De Langhe 1986). Edible parthenocarpic diploids are however cul-
tivated in certain regions such as Tanzania (Simmonds 1962) where the Mchare (or
Mshale) diploids are preferred for the unique texture characteristics of the fruit.
Generally, modern classication systems of banana tend to follow Simmonds
and others (Simmonds and Shepherd 1955; Stover and Simmonds 1987) and are
based on ploidy status and the relative contribution of the two genomes. Simmonds
(1962, 1966) suggested that the formal Latin nomenclature should be replaced by a
ploidy-based nomenclature in which the cultivar is referred to by the genus and a
genomic grouping (e.g., Musa, AAA Group, “Gros Michel”). Cultivars are placed
in higher-level groupings based on the number of chromosomes and the species
that contribute to their genetic makeup (AA, BB, AAA, AAB, and ABB) (Karamura
etal. 2012). Simmonds and Shepherd (1955) utilized 15 taxonomic characters spe-
cic to M. balbisiana and M. acuminata to assign cultivars to groups, and this clas-
sication scheme has been periodically updated (Stover and Simmonds 1987).
Below the level of group, cultivars are assigned to clusters of subgroups that
are characterized by a representative member. For example, “Cavendish” and
“Gros Michel” are considered separate subgroups under the AAA grouping along
with several mutants and variants derived from these economically important cul-
tivars. The grouping (AAA) also includes all of the economically important East
African Highland cooking bananas. While this classication system may be
7 Bananas andPlantains (Musa s pp.)
222
convenient, it appears to lack hierarchical, biological, or economic signicance,
for example, Mysore, Pome, and Plantain are all recognized subgroup clusters
within the AAB group, but they are utilized for different purposes, and subsequent
results of molecular and morphological diversity studies suggest that they are
genetically distinct and likely have arisen from dissimilar parentage (De Langhe
etal. 2010; Christelová etal. 2017).
Likely, cultivars within the AAA, AAB, and ABB groupings arose from multi-
ple hybridization events followed by subsequent backcrossing to various AA, BB,
and AB progenitors which results in an unequal chromosomal allocation at meiosis
(De Langhe etal. 2010). It has been suggested that this phenomenon could explain
the unequal and nonadditive chromosomal complementation which has been
observed among interspecic hybrids (d’Hont et al. 2000). If indeed cultivars
within groupings arose from multiple hybridization events, it suggests that classi-
cation may be more dependent on specic diploid progenitors than on traditional
groupings based on ploidy. Morphological characteristics and nuclear and cyto-
plasmic molecular markers have been used to differentiate the progenitor M. acu-
minata diploids into several subspecies that correspond to specic geographic
ranges from mainland Asia to the archipelagoes of Indonesia, New Guinea, and the
Philippines (Hippolyte et al. 2012; Carreel et al. 2002; Perrier et al. 2011).
Currently, there are eight recognized diploid AA subspecies that include roughly
from West to East: M. acuminata ssp. burmannica, M. acuminata ssp. siamea, M.
acuminata ssp. truncata, M. acuminata ssp. malaccensis, M. acuminata ssp. zeb-
rina, M. acuminata ssp. microcarpa, M. acuminata ssp. errans, and Musa acumi-
nata ssp. banksii. The subspecies have contributed important diploid parents to
modern breeding programs, but much work remains toward evaluating and pre-
serving germplasm that has not been readily accessed due to logistic or political
reasons in past collecting expeditions.
Considerable efforts have been made over the past few decades to preserve, char-
acterize, and provide access to genetic resources of Musa. Banana germplasm for
use in breeding is distributed through the Biodiversity International Musa
Germplasm Transit Centre which oversees more than 1500 accessions. The center
secures available banana germplasm for long-term conservation and holds the col-
lection in trust for the benet of future generations under the auspices of the Food
and Agriculture Organization of the United Nations. The conserved germplasm is
placed in the Multilateral System of Access and Benet Sharing of the International
Treaty on Plant Genetic Resources for Food and Agriculture. All accessions have
been indexed, conserved invitro (Van den houwe etal. 1995), and most stored under
cryopreservation (Panis etal. 2005). Characterization of germplasm occurs in both
eld trials and at the molecular level. Passport and characterization data is freely
available through the Musa Germplasm Information System (MGIS) (https://www.
crop-diversity.org/mgis/).
A. Brown et al.
223
7.3 Banana Breeding Objectives
The primary objective of most banana breeding programs is the uniform production
of large bunches that meet the regional qualitative and quantitative demands of grow-
ers. These demands include superior fruit quality, high suckering ability, short stat-
ure, and enhanced root systems that provide effective soil anchorage and efcient
uptake of water and minerals. Other agronomic traits such as photosynthetic ef-
ciency and rapid cycling are also important breeding objectives for increased yield.
The relative importance of these objectives varies across geographic regions, among
subgroups of banana, and with the intended nal use of the product. In recent years,
the anticipated and realized threats of pests and diseases have resulted in increased
emphasis placed on identifying and utilizing improved sources of host- plant resis-
tance to pests and diseases, particularly in regard to the Sigatoka complex, multiple
races of Fusarium wilt, bacterial wilt, bunchy top, nematodes, and weevils.
In Uganda, banana breeding focuses largely on the improvement of East African
Highland (cooking) bananas (EAHBs) (AAA). The expected yield and plantation life
of these bananas has signicantly declined, in no small part due to pests (such as
banana weevils and nematodes) and diseases (including black Sigatoka and bacterial
wilt). Some of the key breeding objectives by the National Banana Research Program
of the Uganda National Agricultural Research Organization (NARO), in partnership
with the International Institute of Tropical Agriculture (IITA), have been to identify
and integrate host-plant resistance to the Sigatoka complex, weevils, and nematodes
from wild diploid progenitors into elite EAHB backgrounds. A generalized criterion
for selection of EAHB based on agronomic traits is presented in Table7.1.
Table 7.1 Characteristics of the ideotype of East African Highland cooking bananas
Trait Description
Yield potential >25t/ha/year
Bunch weight >15kg
Plant height <3m
Time of owering 210–270days
Time of bunch maturity 90–120days
Number of hands 8–12/bunch
Number of ngers 100–190/bunch
Fruit nger circumference 10–15cm
Fruit nger length 13–20cm
Suckering ability 75% follower sucker growth at harvest
Root system Vigorous (fast growing, deep, and branched)
Bunch orientation Pendent
Reaction to prevalent diseases Resistant to the black Sigatoka complex and bacterial
wilt
Reaction to prevalent pests Resistant to weevils and nematodes
Reaction to drought stress Resistant/tolerant
7 Bananas andPlantains (Musa s pp.)
224
7.4 Constraints toBanana Breeding
As previously discussed, the greatest constraint to banana genetic improvement is
the limited production of viable seeds due to polyploidy, female sterility, and
other factors affecting seed production in triploid and diploid banana. Female
sterility has been intensied as a consequence of human selection for partheno-
carpy in banana. Simmonds (1962) rst suggested that continuous clonal propaga-
tion of diploids has led to an accumulation of structural chromosomal changes
(translocations, inversions, and other events) that restrict normal meiosis and pol-
len fertility and reduce expected recombination. Specic abnormalities such as
translocations have been noted in the diploids “Pisang Lilin” and “Pisang Jari
Buaya,” but the extent to which this phenomenon occurs throughout Musa is still
not well understood. Adeleke etal. (2004) observed that in general, a higher
incidence of univalent formation was related to low pollen fertility in both diploids
and triploids. Sterility in plantains and EAHB triploid bananas has been associ-
ated with meiotic irregularities and uneven number of chromosomes, as well as to
environmental factors and to inuences of individual genotypes (Swennen and
Vuylsteke 1993; Ssebuliba etal. 2000). Seed yield is inuenced by time of polli-
nation, environmental conditions, genetic variation in female fertility, differences
observed among pollinations made between the basal and distal hand, and variation
associated with the relative contributions of the acuminata and balbisiana
genomes (Simmonds 1962). Sathiamoorthy and Rao (1980) observed increased
seed set with proportional contributions from the balbisiana genome and specu-
lated that the factors for seed sterility have accumulated preferentially in the M.
acuminata genome (Simmonds 1962). The use of embryo rescue has signicantly
improved seed germination with observations of up to 30% increases in viable
embryos (Swennen and Vuylsteke 1993).
A further complication associated with improvement of cultivated bananas is
that the highly heterozygous state of the parents results in extremely variable
progeny that makes predictions of progeny performance on the basis of parental
phenotypes unreliable (Ortiz 2000). The progeny from crosses can include mix-
tures of ploidy levels and sometimes aneuploids. Oselebe etal. (2006) reported
that while progeny of 2×–2× crosses were almost exclusively diploid (99.7%),
those of mixed ploidy crosses tended to include individuals that varied in their
chromosomal compliment. The same study observed that the direction of the
cross impacted results. When the diploid is used as the maternal parent in a 2×–4×
cross, over 96% of progeny are diploids, but when the tetraploid is used as the
maternal parent (4×–2×), the observed progeny is predominately triploid (94%)
with varying degrees of other ploidy levels between the diploid and the pentaploid
observed. While these mixed ploidy progenies provide a mechanism for enhanc-
ing genetic diversity and recombination, they also necessitate the use of early
screening of ploidy levels.
A. Brown et al.
225
7.5 Breeding Strategies
Plant breeding provides one of the highest returns of investment in agricultural
research, and while banana production has beneted from these investments, few
banana and plantain cultivars acceptable to farmers and consumers were produced
prior to the 1980s (Roux 2001). Triploid bananas are preferred by growers as they
commonly display the most advantageous combination of fruit and vegetative char-
acters (De Langhe 1986; Stover and Simmonds 1987), but it was generally assumed
that these triploid cultivars (such as “Cavendish”) were effectively sterile (Stover
and Buddenhagen 1986). Persistent efforts, however, from multiple breeding pro-
grams including IITA and the Honduran Foundation for Agricultural Research
(FHIA) have demonstrated that viable embryos could be produced from what were
previously considered recalcitrant triploids by making hybridizations with selected
pollen of diploid banana (Aguilar-Moran 2013). FHIA successfully produced 40
viable tetraploid embryos utilizing “Cavendish” as a female, although this effort
required an almost Herculean task that included the pollination of over 20,000
bunches. By the end of the twentieth century, efforts to improve banana focused
primarily on the use of improved diploid and synthesized tetraploid gene pools to
develop secondary triploids of bananas and plantains (Fig.7.1).
Under this breeding scheme, the identication of improved diploids that provide
donor traits of interest and cultivated triploids with superior quality characteristics
takes on a vital role in synthesizing superior secondary triploids. Conventionally, it
was assumed that when crosses were made between male diploids and female trip-
loids to obtain tetraploid progeny, all three sets of maternal chromosomes were trans-
Generatio
n
0
X
3x
0
2x
0
Generatio
n
1
(Pri
mary hybrids)4x
1
X2x
1
Improved diploi
d
Generation
2
(Seconda
ry triploid) 3x
2
Fig. 7.1 The scheme of the banana breeding process whereby initial crosses are carried out
between triploid landrace (2n=3x) and diploid (2n=2x)
7 Bananas andPlantains (Musa s pp.)
226
ferred intact to the tetraploid offspring with recombination only occurring as a result
of the contribution of diploid male parent (Dodds 1943). Vuylsteke etal. (1993),
however, noted that tetraploid progeny from such crosses displayed variation in dis-
ease resistance, morphological traits, and growth and yield parameters that were
inconsistent with this hypothesis. It was further suggested that segregation and recom-
bination during modied megasporogenesis leading to the formation of 2n eggs in the
triploid parent perhaps better explained the observed results. With the advent of new
genomic resources and tools, this phenomenon needs to be further investigated in
order to better understand the extent of sexual recombination in banana breeding.
This breeding strategy has been adopted by multiple programs including FHIA
in Honduras which has produced a generation of acceptable tetraploids that are still
currently being used in breeding (Rowe and Rosales 1993). An example includes
FHIA 21, a tetraploid derived from AVP-67 French plantain that is still being uti-
lized in plantain improvement. IITA has also utilized this approach successfully to
introgress alleles for resistance/tolerance to key pest and diseases in high-yielding
hybrids derived from preferred plantain cultivars (Ortiz etal. 1995; Tenkouano and
Swennen 2004; Vuylsteke etal. 1993). Plantain hybrid releases include PITA 14 and
PITA 17 (primary tetraploids) and more recently PITA 21, PITA 23, and PITA 24
(secondary triploids) all derived from three seed-fertile triploid French plantains
Obino L’ewai, Bobby Tannap, and Mbi Egome. Their common attributes include
BLS resistance/tolerance and good bunch characteristics (Tenkouano etal. 2011)
and early suckering (Vuylsteke etal. 1993). These IITA hybrids have currently been
distributed to ten countries in Africa and three countries in Central and South
America for evaluation and adoption.
At NARO, tetraploids (AAAA) were synthesized from EAHBs (AAA) by cross-
ing to the wild-seeded, fertile male parent Calcutta 4 (AA) that is used by many
programs as a source of resistance to multiple pests and diseases. A number of these
tetraploids developed such as 365K-1, 1201K-1, 917K-2, 660K-1, 1438K-1, and
222K-1 25 were fundamental in the development of 27 NARITA triploid banana
hybrids by NARO and IITA (Tushemereirwe etal. 2015). These banana hybrids
were selected from early evaluation trials based largely on resistance to black
Sigatoka and bunch size and subsequently advanced to the preliminary yield trials
in Uganda. NARITAs are currently under evaluation for agronomic, sensory, pest,
and disease resistance traits in multi-environment trials in Uganda and Tanzania.
An alternative breeding scheme has been suggested by Vakili (1968) and involves
the polyploidization of diploid hybrids or cultivars through the use of colchicine or
oryzalin to obtain tetraploids for crossing with 2× lines to generate triploids. This
approach is currently being pursued by multiple breeding programs and shows con-
siderable promise (Bakry etal. 2009). According to Tenkouano etal. (2011), the two
schemes conform to differences in breeding philosophies. The former can be viewed
as evolutionary breeding as it attempts to mimic the developmental pathway of
Musa by crossing female triploid landraces to diploid accessions of M. acuminata
or M. balbisiana, while the latter can be considered reconstitutive breeding as it
utilizes the most likely diploid ancestors or relatives of triploid landraces for chro-
mosome doubling to create improved triploids.
A. Brown et al.
227
7.5.1 Development ofImproved Diploids
Regardless of the breeding scheme, the narrow genetic variability and limited
fertility among cultivated triploid bananas make diploid bananas vital to genetic
improvement. A number of fertile improved diploids with varying degrees of dis-
ease resistance have been released by IITA and FHIA (Tenkouano etal. 2003; Rowe
and Rosales 1993). Diploid improvement has almost exclusively been through the
use of M. acuminata cultivars such as “Calcutta 4” (M. acuminata), a source of
resistance to the Sigatoka complex, yellow Sigatoka, fusarium wilt, banana weevil,
and burrowing nematodes (Ortiz 2015). Decades of breeding utilizing this material
have resulted in the production of improved diploid lines which combine disease/
pest resistance, short stature, and interesting bunch characteristics (Tenkouano etal.
2003; Krishnamoorthy and Kumar 2005). Developing further improved diploids
that possess multiple sources of resistance, while preserving the quality characteris-
tics of preferred triploid would greatly increase the efciency of breeding efforts
that are hindered by the constraints previously described.
M. balbisiana diploid cultivars have made limited contributions to breeding due to
the presence of endogenous banana streak virus (eBSV) sequences originating from
the B genome that are activated under the appropriate conditions (Iskra- Caruana etal.
2014). Progenies of interspecic acuminata/balbisiana hybridizations have often
been associated with the occurrence of banana streak disease, and this has resulted in
the underutilization of the B genome which could be an important source of drought
tolerance and resistance alleles not found in M. acuminata (Bakry et al. 2009).
Recently, Noumbissie etal. (2016) reported segregation of the eBSV sequences in the
progeny of crosses between the tetraploid hybrid CRBP 39 (+eBSV) and the AA
male parent Pahang (−eBSV), from which resulted triploid eBSV-free offspring.
Umber etal. (2016) also documented the successful creation of diploids free of eBSV
alleles from M. balbisiana diploids suggesting that recombination between M. acu-
minata and M. balbisiana can be accomplished for improvement of both cultivated
banana and plantain without concern of introducing banana leaf streak.
7.5.2 Breeding Methodology and Evaluation ofHybrids
Production of viable seed through hybridization is critical for the success of breed-
ing and is dependent on residual fertility of triploid cultivars. Practical aspects of
articial hybridization have been described by Tenkouano et al. (2011).
Hybridizations are made through manual pollinations in the early hours of the
morning when pollen availability is not a limiting factor. It can take up to several
months to obtain seeds from a desired cross, and production of seeds is generally
poor and has been reported in the range of 0.3–21.7 seeds per bunch (Swennen and
Vuylsteke 1993) (Fig.7.2). Seeds obtained from crosses also germinate poorly, and
it is standard practice by most programs to recover hybrids through invitro culture
(Bakry 2008) (Fig.7.3).
7 Bananas andPlantains (Musa s pp.)
228
Bunches are harvested prior to physiological maturity, generally when the rst
signs of yellow color are observed in the distal ngers. Bunches are left to ripen in
protected sheds, and seeds are extracted and surface-sterilized for embryo culture to
avoid seed/embryo desiccation. Embryos are extracted from dissected seeds under
aseptic conditions and cultured on articial culture media (Bakry etal. 2009; Uma
et al. 2011). Longitudinal excisions are made on the seed to expose the embryo
Fig. 7.2 Male banana owers open in the evening and are ready for pollination early in the
morning
Fig. 7.3 Bract is pulled back and pollen applied directly to the receptive female owers
A. Brown et al.
229
beneath the micropyle. The embryo is placed on sterile culture medium and incubated
in the dark for germination. Embryos typically germinate between 5 and 20days after
which time they are transferred to an environment with appropriate light and dark
cycles for shoot and root development. Well-developed seedlings can then be cloned
to replicate one to four rooted plantlets or transferred to a screenhouse for condition-
ing prior to eld evaluations. More research on pollen production, pollen tube growth,
and embryo viability is required to better understand issues associated with poor seed
production and to optimize conditions that will lead to better seed yield (Uma etal.
2011). In particular, detailed knowledge of oral biology and seed development is
crucial for recovery of progeny from crosses (Fortescue and Turner 2011).
In the eld, new hybrids are subjected to early evaluation trials (EETs) with lim-
ited replication (one to ve plants). EETs are observed generally for two cycles
during which a few simply inherited traits such as bunch size and orientation,
Sigatoka resistance, seed production, and ploidy level are evaluated. Plants that
show promise in EETs are further evaluated in preliminary yield trials (PYTs)
where replicated clones of selected hybrids are evaluated over two cycles. PYTs
involve more detailed evaluation for additional, complex traits such as yield and
disease resistance. Finally, superior-performing plants from PYTs are cloned in sig-
nicant numbers to allow for multi-locational evaluation trials (MET) that often
include direct input from farmers (Tenkouano etal. 2011). In theory, this process
can take a minimum of 7years to produce a superior banana hybrid, although in
practice this time frame is often exceeded.
7.6 Applied Biotechnology
7.6.1 Molecular-Assisted Breeding
Due to the breeding constraints previously discussed, the use of molecular markers
holds considerable promise in improving the efciency of banana breeding but is
currently not routinely used in most breeding programs. Efforts toward the develop-
ment and use of molecular markers have been greatly facilitated by the recent
release and renement of the draft genomic sequence of the double haploid M.
acuminata cultivar “Pahang” (A genome) (D’Hont etal. 2012; Martin etal. 2016)
and a draft sequence of M. balbisiana “Pisang Klutuk Wulung” (B genome, Davey
etal. 2013). Multiple transcriptome data sets have also been published (Li etal.
2013; Wang etal. 2012), and of these publications, over 45,000 expressed sequence
tags, and 34,000 annotated genes associated with Musa are currently available
through NCBI-EST database. These genomic resources have contributed to the
availability of multiple classes of markers summarized in Tables 7.1 and 7.2.
Molecular markers provide genetic “landmarks” for tagging important traits in plant
breeding, conducting linkage analysis and estimates of genetic diversity, facilitating
gene introgression through marker-assisted studies, providing validation of taxon-
omy and cultivar identication, and estimating evolutionary and speciation events.
7 Bananas andPlantains (Musa s pp.)
230
Table 7.2 A summary of molecular markers utilized in banana research and their research applications
Marker application Marker type Reference
Molecular systematics Isozymes, SSR, DArTs, RFLP and ITS Perrier et al. (2011), Christelová et al. (2017), and Simmonds (1966)
Genetic diversity studies RAPD, SSR, AFLP and MSAP Karamura et al. (2016), Kitavi et al. (2016), Nyine and Pillay (2011), Opara
et al. (2010) Wang et al. (2007), Noyer et al. (2005), Creste et al. (2004), Ude
et al. (2003), Ude et al. (2002), Pillay et al. (2001), Crouch et al. (2000), and
Crouch et al. (1999)
Genome characterization RAPD, RFLP, ITS, dCAP and IRAP
Cultivar identication and
pedigree tracking
Isozymes, RFLP, SSR, RAPD,
EST-SSR and ISSR
Mbanjo et al. (2012), Hippolyte et al. (2012), and Raboin et al. (2005)
Linkage analysis Isozyme, RAPD, RFLP, AFLP, SSR
AS-PCR and DArTs
Mbanjo et al. (2012), Hippolyte et al. (2010) and Fauré et al. (1993)
Genome-wide association
studies and marker-assisted
selection
Isozymes, dCAP and SNP Sardos et al. (2016), Umber et al. (2016), and Noumbissié et al. (2016)
A. Brown et al.
231
7.6.1.1 Association ofMolecular Marker withImportant Genes
The tagging of signicant genes contributing to traits of interest with genic or
linked markers allows for screening of plant germplasm at the earliest stages of
development. The association of these markers with important traits can come
through classical linkage or association studies or through candidate gene
approaches that leverage recently available genomic resources. An example of the
candidate gene approach is provided by Emediato et al. (2009) who amplied
homologues of black leaf streak disease resistance genes in Musa through the use
of degenerate primers based on genes from other crops. The study successfully
amplied sequence differences between the diploid M. acuminata cultivars
“Calcutta 4” (resistant) and “Pisang Berlin” (susceptible). This work followed the
earlier identication of 50 distinct tagged nucleotide binding site-leucine-rich
repeat (NBS-LRR) resistance gene analogs in cultivar “Calcutta 4” by Miller etal.
(2008). Wang et al. (2012) used pooled DNA from Fusarium oxysporum f.sp.
cubensis (Foc TR4)-resistant and susceptible cultivars to identify randomly ampli-
ed polymorphic DNA (RAPD) markers that could distinguish between resistant
and susceptible cultivars. Two RAPD markers were converted to sequence charac-
terized amplied region (SCAR) markers which could be amplied in Foc TR4-
resistant banana genotypes (“Williams 8818-1” and Goldnger), but not in ve
tested susceptible banana cultivars. Work on this continues at the national banana
program in Brazil (EMBRAPA) and shows great promise in providing an early
screen for resistance to Foc TR4 (Silva etal. 2016).
As previously discussed, endogenous banana streak virus (eBSV) limits the
extensive use of the B genome in banana breeding, but the tagging of this
sequence has opened possibilities of greater utilization in the future. Lheureux
etal. (2003) mapped the eBSV sequence using amplied fragment length poly-
morphism (AFLP) markers, and Noumbissié etal. (2016) used simple sequence
repeat (SSR) markers and eBSV-specic PCR markers to identify hybrids lack-
ing the eBSV sequence. Umber etal. (2016) successfully identied infectious
and noninfectious BSV alleles using derived cleaved amplied polymorphic
sequences (dCAPS). These studies suggest that these markers can be used early
in the breeding process as diagnostic markers for eBSV-free B genome hybrids
that will greatly enhance breeding efforts. While the progress shown by these
early efforts is promising, markers associated with traits of economic importance
need to be validated in broader germplasm pools over multiple years to ensure
that they will prove to be reliable and stable and that genotypic predictions at the
early stages of screening will be highly correlated with plant phenotypes at full
maturity under eld conditions.
7 Bananas andPlantains (Musa s pp.)
232
7.6.1.2 Linkage, Association Mapping, andGenomic Selection
Genetic linkage maps provide opportunities for gene identication and a mechanism
for understanding the inheritance pattern of both qualitative and quantitative traits.
Mapping requires appropriate plant populations of known structure derived from
parents that differ signicantly in traits of interest, a set of markers segregating in
the given population that provides substantial coverage of all chromosomes, and the
careful collection of phenotypic data from multiple years and preferably locations.
Linkage mapping has not gained signicant practical application in banana breed-
ing. This could be attributed in part to limitations inherent in marker technologies
and analysis (Foolad 2007; Pillay etal. 2012), to previously described chromosomal
abnormalities in banana that inhibit recombination and to contribute ambiguous
assignment of marker location. To date, principally F1 and F2 diploid populations
have been utilized due to difculties associated with developing double haploid or
recombinant inbred lines in banana.
The rst genetic mapping population in banana was reported in 1993 (Fauré
etal. 1993) and consisted of 92 F2 progeny (AA) derived from an F1 hybrid (SFB5)
of the cross “SF265” (CIRAD-IRFA II.04.20.004.020) × “banksii” (CIRAD-
IRFA II.04.01.004.001). Seventy-seven loci consisting of RAPDs, isozymes, and
restriction fragment length polymorphisms (RFLPs) were mapped onto 15 linkage
groups spanning 606cM. Segregation distortion was associated with 36% of the
mapped loci and was biased toward the “banksii” parent. Hippolyte etal. (2010)
published a more saturated map using an F1 diploid “AA” population created from
a cross between “Borneo” and “Pisang Lilin.” The map was constructed using 426
markers (SSR and DArT). Separate maps were constructed for markers that segre-
gated from each of the heterozygous parents, and a synthetic map was constructed
that spanned 11 linkage groups and represented 1197cM.Three regions of this
synthetic map were inconsistent between the two parents and were attributed by
the authors to structural rearrangements. Subsequent mapping projects have also
noted such incongruities, and while these suggestions are supported by cytogenetic
evidence such as multivalent pairing (Shepherd 1999), much work needs to be
done to verify this hypothesis and determine the extent that such phenomenon
occurs across Musa spp. Mbanjo etal. (2012) produced the most recent map utiliz-
ing an F1 population consisting of crosses between 6142-1 × 8075-7 and 6142-
1-S × 8075-7. Two maternal (6142-1 and 6142-1-S) and one paternal (8075-7)
maps were generated using diversity array technology (DArT), SSR, and allele-
specic PCR (AS-PCR) markers. As with other maps, considerable (41%)
segregation distortion was observed at marker loci.
Association mapping has been proposed as an alternative to conventional linkage
mapping. In this strategy, a panel of genotypes from unrelated population (or a
population with known genetic substructure) is utilized to identify associations
between molecular markers that are in linkage disequilibrium with genetic loci
affecting phenotypes. Molecular markers that are distributed throughout the genome
such as single nucleotide polymorphisms (SNP) are preferred for genome-wide
association studies (GWAS). Sardos et al. (2016) demonstrated the technique
by assembling a GWAS panel of 104 AA accessions using 5544 SNP markers
A. Brown et al.
233
derived from genotyping by sequencing (GBS) and publicly available phenotypic
data on parthenocarpy. The study identied 13 genomic regions associated with
parthenocarpy, and multiple candidate genes in these regions corresponded with
putative growth regulators and genes associated with gametophyte development and
female sterility in other plant species.
Genomic selection (GS) is a form of marker-assisted selection that utilizes high-
density molecular markers such as SNPs to provide coverage of the whole genome,
ensuring that all quantitative trait loci (QTL) are in linkage disequilibrium with at
least one marker (Hayes and Goddard 2010). GS estimates the genomic breeding
value of individual genotypes in a large segregating population utilizing one of sev-
eral GS models (Meuwissen etal. 2001). GS is less concerned with the identica-
tion of individual QTL as it is with developing appropriate models to enhance
selection efciency. As the cost of generating marker data becomes increasingly
more affordable, GS has become an attractive alternative to many breeding pro-
grams (Lorenz etal. 2011; Crossa et al. 2010). Currently, efforts are underway to
evaluate GS as a strategy to improve banana by generating appropriate breeding
models for the improvement of EAHB (Nyine etal. 2016).
7.6.1.3 Estimating Genetic Diversity andEvolutionary Events
Estimates of genetic diversity determine to a large extent the potential of plant
improvement that can be anticipated and can also provide guidance to breeders as to
the appropriate parents to use in breeding schemes. Estimates based on phenotypic or
morphological characters have long been used in banana (Karamura 1998), but often
these estimates can be biased by environmental inuences as well as the sometimes
complimentary and polygenic nature of underlying genetic factors. Molecular mark-
ers avoid these issues as they are highly heritable and have supplemented or replaced
the usage of such measurements in most plant species where they are available. In
banana, several classes of molecular markers have been used to estimate diversity
among populations of varying size representing regional collections and breeding
program germplasm. These include RFLPs (Jarret et al. 1993; Bhat et al. 1995),
RAPDs (Bhat etal. 1995; Crouch etal. 2000; Pillay et al. 2001; Ude et al. 2003;
Nyine and Pillay 2011), AFLPs (Ude etal. 2002, 2003; Noyer etal. 2005; Wang etal.
2007; Opara etal. 2010), SSRs (Kaemmer etal. 1997; Crouch etal. 1999; Tenkouano
etal. 1999; Noyer etal. 2005; Creste etal. 2004; Hippolyte etal. 2012; Kitavi etal.
2016; Karamura etal. 2016), sequence-related amplied polymorphisms (SRAPs)
(Wei etal. 2011; Valdez-Ojeda etal. 2014), DArT (Risterucci etal. 2009), and meth-
ylation-sensitive amplication polymorphism (MASP) (Noyer etal. 2005).
Estimates of genetic diversity generated from these studies vary with the class
and number of markers used and with the genotypes selected for inclusion in any
given study, but a few generalized observations can be made: (1) Diversity esti-
mates based on phenotypic measurements are often poorly correlated with molecu-
lar estimates (Crouch et al. 2000); (2) despite considerable phenotypic or
morphological variation among regional Musa landraces, they tend to have limited
genetic variation when assayed with molecular markers. For example, East African
7 Bananas andPlantains (Musa s pp.)
234
Highland bananas (EAHBs) have been classied into ve clades (clone sets) based
on their end use and morphological distinctiveness (Karamura 1998). Studies
focusing on EAHB using both RAPDs and SSR markers, however, found limited
evidence to support the signicant variation either within or between these clades
(Pillay etal. 2001; Kitavi etal. 2016; Karamura etal. 2016). This led to the sugges-
tion that EAHB arose from a single hybridization event that has subsequently been
acted on by a series of somatic mutations and inuenced by natural and directed
selection leading to many distinct cultivars. Presumably, the nite numbers of
markers used are unable to distinguish among the clades. Utilizing different classes
of molecular markers can sometimes reveal variation in populations where it has
not been previously noted. In plantain landraces of West Africa, RAPD, SSR, and
amplied fragment length polymorphism (AFLP) markers displayed few polymor-
phisms (Crouch etal. 2000). However, when HpaII and MspI methylation-sensitive
amplied markers were used, polymorphism (MSAP) proles revealed three clus-
ters (Noyer etal. 2005) and a genetically distinct subset of plantains from Cameroon
(Ude etal. 2003).
Molecular markers have played important roles in determining the evolutionary
history of cultivated banana and establishing links to diploid progenitors. Whether
the breeder utilizes an evolutionary or reconstitutive approach to banana improve-
ment (discussed in a previous section) plays a vital role in effectively combining
novel resistance traits with quality characteristics desired by growers. Perrier etal.
(2011) detail the available molecular, archaeology, genetic, and linguistic evidence
for this important aspect of breeding. Of particular interest to the dessert banana
industry has been the observation that East African diploid bananas appear to have
played an important evolutionary role in the development of “Cavendish” and “Gros
Michel,” the most widely used cultivars that have dominated the banana export
industry over the past century (Raboin etal. 2005; Risterucci etal. 2009).
7.7 Conclusions
While progress has been made toward genetic improvement since the rst formal
programs were established almost a hundred years ago, in some aspects the breed-
ing of banana is still in its infancy when compared to the improvement of other
important staple crops. In no small part, this is due to the physical and reproductive
constraints of the plant itself, but there is room for optimism as these constraints
appear to be neither absolute nor prohibitive. Molecular markers, the availability of
additional genomic resources, and ongoing studies elucidating the oral and repro-
ductive biology of banana hold great promise for the next hundred years of banana
improvement.
Genetic engineering has not been discussed in this chapter, but the early work in
this arena also suggests that it also has the potential to make an important contribu-
tion to Musa improvement through the introduction of genetic factors not found
within cultivated or wild Musa germplasm (Tripathi etal. 2012).
A. Brown et al.
235
Further work is needed on predicting the performance and combining ability of
male and female parents in Musa improvement. Tenkouano etal. (2012) reported
the signicance of additive genetic effects on expression of bunch weight, fruit
lling time, fruit length, plant height, and number of leaves and nonadditive effects
for suckering behavior and fruit circumference in 3× hybrids obtained from plan-
tain derived 4×–2× crosses. They further suggested that maternal general combin-
ing ability (GCA) accounted for the additive genetic variation for plant height and
number of leaves, while paternal GCA effects accounted for fruit lling time,
bunch weight, and fruit length. On the other hand, specic combining ability
(SCA) effects were observed for all traits, except fruit lling time, suggesting that
additional genetic gain could be achieved through recombinative heterosis for such
traits. They concluded that increased bunch weight and faster cycling are inherited
from the 2× male parent, while plant height, number of leaves, and suckering
behavior are inherited from the 4× female parent which should guide parental
selection for 4×–2× crossbreeding. More of this information is needed to ef-
ciently guide the decision- making efforts of breeders and allow them to allocate
limited resources.
Finally, in the popular press, there has been considerable alarm in recent years as
to the future of banana in the face of an evolving pathogen (Foc TR4) that threatens
the production of much of the world’s dessert banana production. In some ways, the
economic damage that this pathogen will likely cause can be viewed as self-inicted
in nature. The export industry has demonstrated an overdependence on monoculture
and complacency in regard to breeding that has signicantly contributed to creating
an environment conducive to the selection and spread of novel pathogenic races.
This is a lesson that should have been learned more than a half century before when
a similar threat was encountered by a different race of the same pathogen. Banana
breeding efforts to improve the industry standard “Gros Michel” were curtailed or
sidelined when a suitable replacement (“Cavendish”) was selected from existing
stock. Hopefully, the current crises will provide an impetus and serve as a reminder
to all that proactive breeding programs are the most efcient and cost-effective
frontline defense against current and evolving threats to production.
Acknowledgments This research was undertaken with the support of the Belgium Government,
IITA, the Bill and Melinda Gates Foundation, the CGIAR Research Program on Roots, Tubers and
Banana (RTB), USAID and HarvestPlus, part of the CGIAR Research Program on Agriculture for
Nutrition and Health (A4NH).
References
Adeleke MT, Pillay M, Okoli BE (2004) The relationships between meiotic irregularities and fer-
tility in diploid and triploid Musa L.Cytologia 69:387–393
Aguilar Morán JF (2013) Improvement of Cavendish banana cultivars through conventional breed-
ing. Acta Hortic 986:205–208
van Asten PJA, Fermont AM, Taulya G (2011) Drought is a major yield loss factor for rainfed East
African highland banana. Agric Water Manag 98:541–552
7 Bananas andPlantains (Musa s pp.)
236
Bakry F (2008) Zygotic embryo rescue in bananas. Fruits 63:111–115
Bakry F, Carreel F, Jenny C, Horry JP (2009) Genetic improvement of banana. In: Jain SM,
Priyadarshan PM (eds) Breeding plantation tree crops: tropical species. Springer, NewYork,
pp3–50
Bhat KV, Jarret RL, Rana RS (1995) DNA proling of banana and plantain cultivars using random
amplied polymorphic DNA (RAPD) and restriction fragment length polymorphism (RFLP)
markers. Electrophoresis 16:1736–1745
Carreel F, De Leon DG, Lagoda P etal (2002) Ascertaining maternal and paternal lineage within
Musa by chloroplast and mitochondrial DNA RFLP analyses. Genome 45:679–692
Christelová P, De Langhe E, Hřibová E et al (2017) Molecular and cytological characterization of
the global Musa germplasm collection provides insights into the treasure of banana diversity.
Biodivers Conserv 26:801–824
Creste S, Neto AT, Vencovsky R etal (2004) Genetic diversity of Musa diploid and triploid acces-
sions from the Brazilian banana breeding program estimated by microsatellite markers. Genet
Resour Crop Evol 51:723–733
Crossa J, de los Campos G, Pérez P etal (2010) Prediction of genetic values of quantitative traits
in plant breeding using pedigree and molecular markers. Genetics 186:713–724
Crouch JH, Crouch HK, Tenkouano A etal (1999) VNTR-based diversity analysis of 2x and 4x
full-sib Musa hybrids. Electron JBiotechnol 2:130–139
Crouch HK, Crouch JH, Madsen S etal (2000) Comparative analysis of phenotypic and genotypic
diversity among plantain landraces (Musa spp., AAB group). Theor Appl Genet 101:1056–1065
D’Hont A, Paget-Goy A, Escoute J et al (2000) The interspecic genome structure of culti-
vated banana, Musa spp. revealed by genomic DNA in situ hybridization. Theor Appl Genet
100:177–183
D’Hont A, Denoeud F, Aury JM etal (2012) The banana (Musa acuminata) genome and the evolu-
tion of monocotyledonous plants. Nature 488:213–217
Davey MW, Gudimella R, Harikrishna JA etal (2013) A draft Musa balbisiana genome sequence
for molecular genetics in polyploid, inter-and intra-specic Musa hybrids. BMC Genomics
14:683
De Langhe E (1986) Towards an international strategy for genetic improvement in the genus Musa.
In: Persley GJ, De Langhe EA (eds) Banana and plantain breeding strategies. Proceedings of an
International Workshop, Cairns, Australia. 1–17 October, 1986. INIBAP, Montpellier, p19–23
De Langhe E, Vrydaghs L, De Maret P etal (2009) Why bananas matter: an introduction to the
history of banana domestication. Ethnobot Res Appl 7:165–177
De Langhe E, Hribova E, Carpentier S etal (2010) Did backcrossing contribute to the origin of
hybrid edible bananas? Ann Bot 106:849–857
Deckers J, Tessera M, Alemu K, Abate T, Swennen R (2001) Ensete. In: Raemaekers RH (ed) Crop
production in tropical Africa. DGIC, Brussels, pp587–591
Dodds KS (1943) The genetic system of banana varieties in relation to banana breeding. Emp
JExp Agric 11:89–98
Dodds KS (1945) Genetical and cytological studies of Musa. VII.Certain aspects of polyploidy.
JGenet 46:161–179
Dumpe BB, Ortiz R (1996) Apparent male fertility in Musa germplasm. HortSci 31:1019–1022
Emediato FL, Nunes FA, Teixeira CC, Passos MA, Bertioli DJ, Pappas GJ, Miller RN (2009)
Characterization of resistance gene analogs in Musa acuminata cultivars contrasting in
resistance to biotic stresses. In: Shu QY (ed) Induced plant mutations in the genomics era.
FAO, Rome, pp443–445
FAO (2014) Banana market review and banana statistics 2012–2013, Rep. I3627E/1/01.14. FAO,
Rome. http://www.fao.org/docrep/019/i3627e/i3627e.pdf
Fauré S, Noyer JL, Horry JP et al (1993) A molecular marker-based linkage map of diploid
bananas (Musa acuminata). Theor Appl Genet 87:517–526
Foolad MR (2007) Genome mapping and molecular breeding of tomato. Int JPlant Genomics
2007:64358
A. Brown et al.
237
Fortescue JA, Turner DW (2004) Pollen fertility in Musa: viability in cultivars grown in southern
Australia. Aust JAgric Res 55:1085–1091
Fortescue JA, Turner DW (2011) Reproductive biology. In: Pillay M, Tenkouano A (eds) Banana
breeding: constraints and progress. CRC Press, Boca Raton, pp305–331
Fortescue JA, Turner DW, Romero R (2011) Romero evidence that banana (Musa spp.), a tropi-
cal monocotyledon, has a facultative long-day response to photoperiod. Funct Plant Biol
38:867–878
Hayes B, Goddard M (2010) Genome-wide association and genomic selection in animal breeding.
Genome 53:876–883
Hippolyte I, Bakry F, Seguin M etal (2010) A saturated SSR/DArT linkage map of Musa acumi-
nata addressing genome rearrangements among bananas. BMC Plant Biol 10:65
Hippolyte I, Jenny C, Gardes L etal (2012) Foundation characteristics of edible Musa triploids
revealed from allelic distribution of SSR markers. Ann Bot 109:937–951
Iskra-Caruana M, Chabannes M, Duroy PO etal (2014) A possible scenario for the evolution of
banana streak virus in banana. Virus Res 186:155–162
Janssens SB, Vandelook F, De Langhe E etal (2016) Evolutionary dynamics and biogeography of
Musaceae reveal a correlation between the diversication of the banana family and the geologi-
cal and climatic history of Southeast Asia. New Phytol 210:1453–1465
Jarret RL, Vuylsteke DR, Gawel NJ et al (1993) Detecting genetic diversity in diploid bananas
using PCR and primers from a highly repetitive DNA sequence. Euphytica 68:69–76
Jones DR (ed) (1999) Diseases of banana, abaca’ and enset. CABI, Wallingford
Kaemmer D, Fischer D, Jarret RL etal (1997) Molecular breeding in the genus Musa: a strong case
for STMS marker technology. Euphytica 96:49–63
Karamura DA (1998) Numerical taxonomic studies of the East African highland (Musa AAA East
Africa) in Uganda. Dissertation, University of Reading
Karamura DA, Karamura EB, Tinzaara W (2012) In: Karamura DA, Karamura EB, Tinzaara W
(eds) The current classication and naming of the East African highland bananas (Musa AAA)
based on Morphological Characteristics in book: Banana cultivar Names, Synonyms and their
Usage. Bioversity International, East Africa, pp 6–23
Karamura D, Kitavi M, Nyine M etal (2016) Genotyping the local banana landrace groups of East
Africa. Acta Hortic 1114:67–74
Kitavi M, Downing T, Lorenzen Jetal (2016) The triploid East African Highland Banana (EAHB)
genepool is genetically uniform arising from a single ancestral clone that underwent population
expansion by vegetative propagation. Theor Appl Genet 129:547–561
Krishnamoorthy V, Kumar N (2005) Preliminary evaluation of diploid banana hybrids for yield
potential, male fertility and reaction to Radopholus similis. Plant Gen Res Newsl 141:39–43
Kumar LP, Selvarajan R, Iskra-Caruana M, Chabannes M, Hanna R (2015) Biology, etiology, and
control of virus diseases of banana and plantain. In: Loebenstein G, Katis NI (eds) Advances in
virus research, vol 91. Academic, Burlington, pp229–269
Lheureux F, Carreel F, Jenny C et al (2003) Identication of genetic markers linked to banana
streak disease expression in inter-specic Musa hybrids. Theor Appl Genet 106:594–598
Li C, Shao J, Wang Y etal (2013) Analysis of banana transcriptome and global gene expression
proles in banana roots in response to infection by race 1 and tropical race 4 of Fusarium oxy-
sporum f. sp. Cubense. BMC Genomics 14:851
Lorenz AJ, Chao S, Franco G etal (2011) Chap. 2: Genomic selection in plant breeding: knowl-
edge and prospects. Adv Agron 110:77–123
Martin G, Baurens FC, Droc G etal (2016) Improvement of the banana “Musa acuminata” refer-
ence sequence using NGS data and semi-automated bioinformatics methods. BMC Genomics
17:1–12
Mbanjo EGN, Tchoumbougnang F, Mouelle AS etal (2012) Development of expressed sequence
tags-simple sequence repeats (EST-SSRs) for Musa and their applicability in authentication of
a Musa breeding population. Afric JBiotechnol 11:13546–13559
Meuwissen TH, Hayes BJ, Goddard ME (2001) Prediction of total genetic value using genome-
wide dense marker maps. Genetics 157:1819–1829
7 Bananas andPlantains (Musa s pp.)
238
Miller RNG, Bertioli DJ, Baurens FC etal (2008) Analysis of non-TIR NBS-LRR resistance gene
analogs in Musa acuminata Colla: isolation, RFLP marker development, and physical map-
ping. BMC Plant Biol 8:15
Noumbissié GB, Chabannes M, Bakry F etal (2016) Chromosome segregation in an allotetraploid
banana hybrid (AAAB) suggests a translocation between the A and B genomes and results in
eBSV-free offsprings. Mol Breed 36:1–14
Noyer JL, Causse S, Tomekpe K etal (2005) A new image of plantain diversity assessed by SSR,
AFLP and MSAP markers. Genetica 124:61–69
Nyine M, Pillay M (2011) The effect of banana breeding on the diversity of East African Highland
banana (Musa, AAA). Acta Hortic 897:225–229
Nyine M, Uwimana B, Swennen R, Batte M, Brown A, Hřibová E, Doležel J(2016) Genomic
breeding approaches for East African Bananas. In: Abstracts of the plant and animal genome
conference XXIV January 08–13, San Diego, CA
Opara UL, Jacobson D, Al-Saady NA (2010) Analysis of genetic diversity in banana cultivars
(Musa cvs.) from the South of Oman using AFLP markers and classication by phylogenetic,
hierarchical clustering and principal component analyses. JZhejiang Univ Sci B 11:332–341
Ortiz R (2000) Understanding the Musa genome: an update. Acta Hortic 54:157–168
Ortiz R (2013) Conventional banana and plantain breeding. Acta Hortic 986:77–194
Ortiz R (2015) Plant breeding in the omics era. Springer, NewYork
Ortiz R, Swennen R (2014) From crossbreeding to biotechnology-facilitated improvement of
banana and plantain. Biotechnol Adv 32:158–169
Ortiz R, Ferris RSB, Vuylsteke DR (1995) Banana and plantain breeding. In: Gowen S (ed)
Bananas and plantains. Springer, NewYork, pp110–146
Oselebe HO, Tenkuoano A, Pillay M etal (2006) Ploidy and genome segregation in Musa breeding
populations assessed by ow cytometry and randomly amplied polymorphic DNA markers.
JAm Soc Hortic 131:780–786
Panis B, Piette B, Swennen R (2005) Droplet vitrication of apical meristems: a cryopreservation
protocol applicable to all Musaceae. Plant Sci 168:45–55
Perrier X, De Langhe E, Donohue M, Lentfer C, Vrydaghs L, Bakry F, Carreel F, Hippolyte I,
Horry J-P, Jenny C, Lebot V, Risterucci A-M, Tomekpe K, Doutrelepont H, Ball T, Manwaring
J, de Maret P, Denham T (2011) Multidisciplinary perspectives on banana (Musa spp.) domes-
tication. PNAS 108:11311–11318
Pillay M, Ogundiwin E, Nwakanma DC etal (2001) Analysis of genetic diversity and relationships
in East African banana germplasm. Theor Appl Genet 102:965–970
Pillay M, Ude G, Kole C (eds) (2012) Genetics, genomics and breeding of bananas. CRC, Boca
Raton
Raboin LM, Carreel F, Noyer JL etal (2005) Diploid ancestors of triploid export banana culti-
vars: molecular identication of 2n restitution gamete donors and n gamete donors. Mol Breed
16:333–341
Risterucci AM, Hippolyte I, Perrier X etal (2009) Development and assessment of diversity arrays
technology for high-throughput DNA analyses in Musa. Theor Appl Genet 119:1093–1103
Roux NS (2001) Mutation induction in Musa. In: Jain SM, Swennen R (eds) Banana improvement:
cellular, molecular biology, and induced mutations. Science, Eneld
Rowe P, Rosale F (1993) Diploid breeding at FHIA and the development of Goldnger (FHIA-01).
InfoMusa 2:9–11
Sardos J, Rouard M, Hueber Y etal (2016) A genome-wide association study on the seedless phe-
notype in banana (Musa spp.) reveals the potential of a selected panel to detect candidate genes
in a vegetatively propagated crop. PLoS One 11:5
Sathiamoorthy S, Rao VNM (1980) Pollen production in relation to genome and ploidy in banana
clones. Proc Nat Semi Banana Prod. Tech., TNAU, Coimbatore, pp65–66
Shepherd K (1999) Cytogenetics of the genus Musa. International Network for the Improvement
of Banana and Plantain, Montpellier
A. Brown et al.
239
Silva PRO, de Jesus ON, Bragança CAD etal (2016) Development of a thematic collection of
Musa spp accessions using SCAR markers for preventive breeding against Fusarium oxyspo-
rum f. sp cubense tropical race 4. Genet Mol Res 15:5017765
Simmonds NW (1962) The evolution of the bananas. Longmans, London
Simmonds NW (1966) Bananas, 2nd edn. Longmans, London
Simmonds NW (1987) Classication and breeding of bananas. In: Persley G, De Langhe E (eds)
Banana and plantain breeding strategies. Proceedings of an International Workshop held at
Cairns Australia, 13–17 October 1986. Austrailian Centre for International Agricultural
Research, Canberra
Simmonds NW, Shepherd K (1955) The taxonomy and origins of the cultivated bananas. JLinn
Soc Lond Bot 55:302–312
Ssebuliba R, Vuylsteke D, Hartman Jet al (2000) Towards improving highland bananas. Uganda
JAgric Sci 5:36–38
Ssesuliba RN, Tenkouano A, Pillay M (2008) Male fertility and occurrence of 2n gametes in East
African highland bananas (Musa spp.) Euphytica 164:153–162
Stover RH, Buddenhagen IW (1986) Banana breeding: polyploidy, disease resistance and produc-
tivity. Fruits 41:175–191
Stover RH, Simmonds NW (1987) Bananas, Tropical agricultural series, 3rd edn. Longmans,
London
Swennen R, Vuylsteke D (1993) Breeding black Sigatoka resistant plantain with a wild banana.
Trop Agric 70:74–77
Tenkouano A, Swennen R (2004) Progress in breeding and delivering improved plantain and
banana to African farmers. Chron Hortic 44:9–15
Tenkouano A, Crouch JH, Crouch HK et al (1999) Comparison of DNA marker and pedigree-
based methods of genetic analysis of plantain and banana (Musa spp.) clones. I.Estimation of
genetic relationships. Theor Appl Genet 98:62–68
Tenkouano A, Vuylsteke D, Okoro Jetal (2003) Registration of TMB2x5105-1 and TMB2x9128-3
diploid banana hybrids with good combining ability, partial resistance to black Sigatoka and
resistance to nematodes. Hortscience 38:468–472
Tenkouano A, Pillay M, Ortiz R (2011) Breeding techniques. In: Pillay M, Tenkouano A (eds)
Banana breeding: constraints and progress. CRC Press, Boca Raton, pp181–202
Tenkouano A, Ortiz R, Vuylsteke D (2012) Estimating genetic effects in maternal and paternal
half-sibs from tetraploid-diploid crosses in Musa spp. Euphytica 185:295–301
Tripathi JN, Muwonge A, Tripathi L (2012) Efcient regeneration and transformation of plantain
cv. ‘Gonja manjaya’ (Musa spp. AAB) using embryogenic cell suspensions. In Vitro Cell Dev
Biol 48:216–224
Tsegaye A, Struik PC (2002) Analysis of enset (Ensete ventricosum) indigenous production meth-
ods and farm-based biodiversity in major enset-growing regions of southern Ethiopia. Explor
Agric 38:291–315
Tushemereirwe W, Batte M, Nyine M, Tumuhimbise R, Barekye A, Tendo S, Talengera D, Kubiriba
J, Lorenzen J, Swennen R, Uwimana B (2015) Performance of NARITA banana hybrids in the
preliminary yield trial for three cycles in Uganda. Banana Technical Report, 35p. www.musalit.
org/seeMore.php?id=15482
Ude G, Pillay M, Nwakanma D etal (2002) Genetic diversity in Musa acuminata Colla and Musa
balbisiana Colla and some of their natural hybrids using AFLP markers. Theor Appl Genet
104:1246–1252
Ude G, Pillay M, Ogundiwin E etal (2003) Genetic diversity in an African plantain core collection
using AFLP and RAPD markers. Theor Appl Genet 107:248–255
Uma S, Lakshmi S, Saraswathi MS etal (2011) Embryo rescue and plant regeneration in banana
(Musa spp.) plant cell. Tissue Organ Cult 105:105–111
Umber M, Pichaut JP, Farinas B etal (2016) Marker-assisted breeding of Musa balbisiana genitors
devoid of infectious endogenous banana streak virus sequences. Mol Breed 36:1–11
Vakili NG (1968) Responses of Musa acuminata species and edible cultivars to infection by
Mycosphaerella musicola. Trop Agric 45:13–22
7 Bananas andPlantains (Musa s pp.)
240
Valdez-Ojeda R, James-Kay A, Ku-Cauich JR etal (2014) Genetic relationships among a collec-
tion of Musa germplasm by uorescent-labeled SRAP.Tree Genet Genomic 10:465–476
Van den Houwe I, De Smet K, Tezenas de Montcel H etal (1995) Variability in storage potential
of banana shoot cultures under medium term storage conditions. Plant Cell Tissue Organ Cult
42:269–274
Vuylsteke D, Swennen R, Ortiz R (1993) Development and performance of Black Sigatoka-
resistant tetraploid hybrids of plantain (Musa spp., AAB group). Euphytica 65:33–42
Wairegi LWI, van Asten PJA, Tenywa MM etal (2010) Abiotic constraints override biotic con-
straints in East African highland banana systems. Field Crop Res 117:146–153
Wang XL, Chiang TY, Roux N et al (2007) Genetic diversity of wild banana (Musa balbisiana
Colla) in China as revealed by AFLP markers. Genet Resour Crop Evol 54:11251132
Wang W, Hu Y, Sun D etal (2012) Identication and evaluation of two diagnostic markers linked
to Fusarium wilt resistance (race 4) in banana (Musa spp.) Mol Biol Rep 39:451–459
Wei JY, Liu DB, Wei SX etal (2011) Analysis of genetic diversity in banana cultivars (Musa spp.)
using sequence-related amplied polymorphism markers. Acta Hortic 897:263–265
A. Brown et al.