Scientia Horticulturae 214 (2017) 91–98

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Scientia Horticulturae
journal homepage: www.elsevier.com/locate/scihorti

Review

Genetically modified bananas: To mitigate food security concerns

Siddhesh B. Ghag a , Thumballi R. Ganapathi b,∗
a
UM-DAE Centre for Excellence in Basic Sciences, Kalina, SantaCruz, Mumbai 400 098, India
b
Plant Cell Culture Technology Section, Nuclear Agriculture & Biotechnology Division, Bhabha Atomic Research Centre, Trombay, Mumbai 400 085, India

a r t i c l e i n f o a b s t r a c t

Article history: Banana and plantains are one of the world’s most important food crops and widely consumed by people
Received 6 June 2016 of all age groups. Bananas are a rich source of carbohydrates, important vitamins like vitamin B and C and
Received in revised form 1 November 2016 minerals like potassium and phosphorous. Banana is an economically important cash crop as it fetch large
Accepted 20 November 2016
revenue share in the domestic and international market. However, most of the production is consumed
by the domestic population as it serves as the staple food for them. Bananas are vulnerable to both biotic
Keywords:
and abiotic stress factors which limits their production. Improvement of this crop to enhance the nutrient
Transgenics
quality and better adapt to the changing environmental conditions and to produce new disease resistant
Bananas
Crop improvement
varieties is essential. Genetic engineering of banana is considered a perfect alternative for improvement
Genetic transformation of sterile cultivars or ones which are not amenable to traditional breeding methods Several successful
Abiotic stress attempts have demonstrated the strength of this technology in developing abiotic stress tolerance and
Biotic stress disease resistant transgenic banana varieties. Only few of the Genetically Modified (GM) bananas have
Biofortification qualified for field studies and some are currently undergoing nutritional human trials. GM bananas aim
Fusarium wilt to increase productivity and nutritional value and so could effectively contribute towards food security
Vitamin A in the near future.
© 2016 Elsevier B.V. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91
2. Conventional versus biotechnological banana improvement approaches . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92
3. Modern banana tissue culture and genetic transformation methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92
3.1. Abiotic stress resistant GM bananas . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92
3.2. Biotic stress resistant GM bananas . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
3.3. Biofortified GM bananas . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95
4. Present challenges and future prospects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95
Conflict of interest . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 96
Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 96
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 96

1. Introduction
about 106.7 million tonnes and India is the largest producer with
Banana and plantains the major staple food crops in the devel- a production of about 29.7 million tonnes (FAO, 2013-14). Many
oping countries are widely cultivated in tropical and subtropical cultivars have poor yield but are still in cultivation because of their
regions across the world. The plant considered as a remunerative quality and apart from being eaten raw and cooked, bananas are
cash crop yield satisfactory returns to the growers and consid- also used in making beer, edible dried chips and flour. Though
ered as the developing world’s most important food crop after many varieties are grown in India, only a few are grown com-
rice, wheat and maize. The annual world production of banana is mercially which include Grand Nain, Robusta, Dwarf Cavendish,
Nendran, Rasthali as the major varieties. The yield is more for
Cavendish bananas compared to Rasthali and Nendran. Indian
∗ Corresponding author. states of Maharashtra and Gujarat together contribute 27%, where
E-mail addresses: siddhesh.ghag@cbs.ac.in (S.B. Ghag), trgana@barc.gov.in the conditions for banana cultivation are favorable (http://nhb.
(T.R. Ganapathi). gov.in/report files/banana/BANANA.htm). The other significantly

http://dx.doi.org/10.1016/j.scienta.2016.11.023
0304-4238/© 2016 Elsevier B.V. All rights reserved.
92 S.B. Ghag, T.R. Ganapathi / Scientia Horticulturae 214 (2017) 91–98

banana growing states in India are Tamil Nadu, Kerala and Kar- envisaged recently with a few publications on disease resistance
nataka. Among numerous varieties, Cavendish bananas constitute and environmental stress tolerance (Paul et al., 2011; Shekhawat
most of the total world export trade of 15 million tones that et al., 2011; Shekhawat et al., 2012; Ghag et al., 2012; Sreedharan
amounts to about US$ 4.8 billion for exporting countries. Banana is et al., 2012). The main advantage being the transfer of useful trait/s
a staple food in some parts of the world and thus a viable crop for to a cultivar which has most of the desirable characters as possible.
biofortification. Banana cultivation is threatened by both biotic and Also the leading commercially grown varieties can be engineered
abiotic factors which limit its production potential. As such banana for multiple stress tolerance, delayed fruit ripening and nutritional
plants are sensitive to low temperatures, drought and salinity caus- quality improvement. Since most of the cultivated bananas are
ing significant losses. Moreover, they are host to major debilitating triploids with very low pollen fertility, apprehensions regarding
diseases such as Fusarium wilt, sigatoka disease, banana bunchy top transgene flow to the wild cultivated bananas are negligible.
disease and Moko disease (Jones, 1999). These diseases can spread
rapidly to an extent of causing epidemics (mostly due to mono-
3. Modern banana tissue culture and genetic
culture practice) and capable to inflict famine conditions in major
transformation methods
banana growing regions. Improvement of banana through genetic
engineering ensures sustainable production.
Conventionally banana is propagated by vegetative suckers. One
Present day high throughput technologies has allowed simul-
of the main limitations with this method is the availability of
taneous investigation of thousands of genes, transcripts, proteins,
uniform age and size suckers in large numbers for a given elite vari-
and metabolites involved in biotic and abiotic stress response.
ety/cultivar. In vitro micropropagation of banana developed for the
Genome and transcriptome sequencing of different banana culti-
last three decades are now considered well established (Vuylsteke
vars have been carried out in recent past which provides valuable
and De Langhe, 1985; Wong, 1986; Vuylsteke, 1989; Arias, 1992).
resource information for banana improvement (D’Hont et al., 2012;
Now, protocols for large scale tissue culture propagation of banana
Li et al., 2013; Bai et al., 2013; Martin et al., 2016; Wu et al.,
using shoot tip cultures have been established (Saraswathi et al.,
2016). The enormous data generated in these sequencing reactions
2016). However a few banana cultivars are recalcitrant to tissue
provide insight in studying novel genes, differential expression pro-
culture and transformation protocols. Further, the protocols estab-
files, comparative genomics study, evolutionary genomics research
lished are also genotype specific. In the last two decades banana
and polymorphisms which can be exploited for banana breeding
biotechnologists have put in considerable efforts in optimizing
and/or transgenic procedures. Transgenic banana offer environ-
parameters for banana tissue culture and regeneration in differ-
mental benefits by reduced spraying of pesticides on disease and
ent cultivars (Ganapathi et al., 2001a, 2001b; Khanna et al., 2004;
pest resistant plants, increased productivity on the available arable
Tripathi et al., 2005). Many of the private commercial tissue culture
land and combating climate change through abiotic stress tolerant
laboratories are producing millions of plantlets and the farmers
plants. This review intends to showcase most of the studies demon-
are reaping the benefits from this development. However, the
strating the potentiality of transgenic banana in tackling production
shoot tip culture method of regeneration is not suitable for genetic
constraints and ensure food security and sustainability.
engineering as there is a problem of chimeras (May et al., 1995).
Alternatively, ECS cultures circumvent this problem as single cells
transformed are regenerated into plantlets (Fig. 1). Various explants
2. Conventional versus biotechnological banana
like immature male/female flower buds, scalps and shoot tip cul-
improvement approaches
tures have been used to initiate ECS in a few of the commercial
cultivars (Escalant and Teisson, 1989; Escalant et al., 1994; Dhed’a
Despite their importance banana and plantains are one of the
et al., 1991; Ganapathi et al., 1999, 2001b; Grapin et al., 2000; Ghosh
least genetically improved crops, and production is based on the
et al., 2009). However difficulties in obtaining ECS from several elite
cultivars derived from few collections which are domesticated
varieties have limited the scope for improvement and conservation
from ages and undergone numerous complex hybridization events
of these important landraces. In vitro development of embryogenic
(Heslop-Harrison and Schwarzacher, 2007). Banana breeding has
callus and ECS requires appropriate and precise cocktail of hor-
been considered difficult due to their long life cycle, triploidy
mones which one needs to optimize for each variety separately.
and parthenocarpic fruit development in most of the edible culti-
Agrobacterium-mediated genetic transformation is the method
vars. Conventional breeding is complex, time consuming and their
of choice used to transform banana cells (May et al., 1995;
results are often not satisfying. However few institutes such as FHIA
Ganapathi et al., 2001b; Khanna et al., 2004; Tripathi et al., 2005;
(Honduras), EMBRAPA (Brazil), CARBAP (Cameroon) NRCB (India)
Huang et al., 2007; Ghosh et al., 2009). However, few research
and IITA (Africa) are breeding banana varieties to obtain triploids
groups employed biolistic/microprojectile bombardment method
and tetraploids with improved characteristics (Jacome et al., 2002;
to deliver foreign gene into banana tissues. Becker et al. (2000)
Bakry et al., 2009). Somaclonal variations induced by repeated
and Matsumoto et al. (2000) bombarded ECS cultures of Cavendish
micropropagation-selection cycles have also been attempted in
banana (Musa spp. AAA group) cv ‘Grand Nain’ or ‘Maçã’ banana
banana to develop resistance to Fusarium wilt (Hwang and Ko,
(Musa sp. AAB group, Silk) respectively to develop stable transfor-
2004). Genetic engineering of bananas acquire special impor-
mants.
tance as useful characters can be introduced relatively rapidly into
elite cultivars without compromising their fundamental genetic
makeup. Establishment of a cell suspension culture based protocol 3.1. Abiotic stress resistant GM bananas
for genetic transformation of banana (Côte et al., 1996; Ganapathi
et al., 2001b; Strosse, 2003; Strosse et al., 2006; Khanna et al., Climate change challenges the optimum productivity of crops
2004, 2007; Chong-Pérez et al., 2012) opened up opportunities in tropical regions. Banana is an evergreen and shallow rooted crop
for banana improvement using genetic engineering techniques. A and requires large quantity of water throughout its life cycle. Due to
high throughput regeneration and transformation for a few cul- climate change there is an increase in temperature which critically
tivars has also been developed recently by Tripathi et al. (2015). disturbs plantations (Gornall et al., 2010). Further, higher temper-
Embryogenic cell suspensions (ECS) and Agrobacterium tumefa- ature causes increased demand for water for cultivation leading to
ciens mediated transformation are well established for few banana droughts. Although warmer temperatures will favor increased area
cultivars and hence improvement using this technique has been under cultivation for banana, the corresponding rise in drought
 S.B. Ghag, T.R. Ganapathi / Scientia Horticulturae 214 (2017) 91–98 93

Fig. 1. Genetic transformation of banana embryogenic cell suspension cultures developed from immature male flower buds. (A) Freshly harvested male flower buds (B)
Immature male flower whorls cultured on medium for embryogenic callus (C) Embryogenic cell suspension cultures developed from embryogenic callus (D) Embryogenic
cells observed under high power of microscope showing dense cytoplasm (E) Embryos developed from the cells on the embryo induction medium (F) Germination of embryos
into plantlets (G) Multiple shoot formation from germinating somatic embryos for clonal propagation (H) Each shoot rooted separately on root induction medium (I) Growing
the plants up to maturity in the experimental field (J) Full grown banana plant showing fruit bunch and male flower bud which can be further utilized for multiplication.

Table 1
List of abiotic stress tolerance genes identified in banana.

Target gene Transgenic Source of gene Stress tolerance References

plant/Cultivar

Musa-DHN-1 Musa spp. Rasthali Musa spp. Karibale Mothan Drought, salt Shekhawat et al. (2011)
MusaSAP1 Musa spp. Rasthali Musa spp. Karibale Mothan Drought, salt, oxidative stress Sreedharan et al. (2012)
MusaPIP1;2 Musa spp. Rasthali Musa spp. Karibale Mothan Cold, salt, drought Sreedharan et al. (2013)
MusaWRKY71 Musa spp. Rasthali Musa spp. Karibale Mothan Salt, oxidative stress Shekhawat and Ganapathi (2013)
MusabZIP53 Musa spp. Rasthali Musa spp. Karibale Mothan Cold, drought, salt Shekhawat and Ganapathi (2014)
MaPIP1;1 Arabidopsis Musa acuminata L. Salt, drought Xu et al. (2014)
AhSIPR10 M. acuminata cv. Matti Arachis hypogaea Salt, drought Rustagi et al. (2015)
MusaPIP2;6 Musa spp. Rasthali Musa spp. Karibale Mothan Salt Sreedharan et al. (2015)
MpMYBS3 Musa spp. cv. Brazil Musa spp. cv. Lingchuan Dajiao Cold Dou et al. (2016)
MusaNAC042 Musa spp. Rasthali Musa spp. Karibale Mothan Salt, drought Tak et al. (2016)

conditions will severely affect yield and productivity. Moreover,
warmer temperature may probably escalate incidence of diseases
such as Fusarium or bacterial wilt. Banana cultivation is also
severely affected by increased salinity and cold temperatures as
temperatures below 12 ◦ C induce chilling damages (Murata, 1969;
Jiang et al., 2004). To develop varieties which can sustain these abi-
otic stress factors, genes involved in abiotic stress tolerance and
underlying mechanism of stress response and tolerance in banana
needs to be studied in depth. As most edible banana cultivars are
parthenocarpic, developing transgenic bananas with the capacity
to grow vigorously under unfavorable climatic and environmen-
tal conditions will be the most ideal approach. No sooner the
plant experience an abiotic stress, it induces a cascade of signaling
events involving transcription factors and downstream effectors
culminating into a tolerance response against the stress agent.
Transcription factors like WRKY (Shekhawat and Ganapathi, 2013),
bZIP (Shekhawat and Ganapathi, 2014), SAP1 (Sreedharan et al.,
2012) and ABA-, stress-, and ripening-induced (ASR) protein (Miao
et al., 2014) have been characterized and shown to be up-regulated
during stress condition. When these transcription factors were
overexpressed in transgenic plants, these plants were capable of
tolerating and surviving under stress condition (Table 1). However,
constitutive overexpression of these transcription factors such as
bZIP53 (Shekhawat and Ganapathi, 2014) may result in growth
abnormality in the transgenic plants as they also play significant
role during growth and development. Identification and overex-
pression of a crucial effector protein involved in stress tolerance
can be an ideal choice. The dehydrin gene MusaDHN-1 identified
94 S.B. Ghag, T.R. Ganapathi / Scientia Horticulturae 214 (2017) 91–98
from banana cv. Karibale Monthan when expressed constitutively in
transgenic banana plants conferred enhanced tolerance to drought
and salt stress conditions without showing any phenotypic abnor-
mality in the plants (Shekhawat et al., 2011). Aquaporins were
shown to be involved in cold, salt and drought stress tolerance
in banana however, their exact role in these stresses needs more
profound research. Genome-wide analysis identified 47 aquaporin
genes in banana categorized into four groups namely the plasma
membrane intrinsic proteins (PIPs), tonoplast intrinsic proteins
(TIPs), nodulin 26-like intrinsic proteins (NIPs), and small basic
intrinsic proteins (SIPs) (Hu et al., 2013). Constitutive overex-
pression of PIPs in transgenic banana plants imparted increased
tolerance to cold, salt and drought stress conditions (Sreedharan
et al., 2013, 2015; Xu et al., 2014). All these studies have been
done under controlled environment of enclosed greenhouse and
even though the results are promising at this level they might not
transfer under natural, uncontrolled field conditions. Therefore it
is essential to test these plants under field conditions to prove its
worthiness.
3.2. Biotic stress resistant GM bananas
Pest and diseases are among the most important reasons for
reduced banana productivity and yield worldwide. Recently, fungal
and viral diseases became the foremost objectives of biotechnolog-
ical strategies to improve this crop. Fusarium wilt disease caused
by Fusarium oxysproum f. sp. cubense (Foc) is the most devastat-
ing fungal disease of banana as it is difficult to control by physical
or chemical methods. Since it’s a soil born pathogen it can be eas-
ily transmitted throughout and between farms. Additionally, the
chlamydospores can remain dormant in the soil for more than 30
years. Furthermore, Foc tropical race 4 (TR4) is of major concern
presently because it threatens the Cavendish industry which pre-
dominates current market. Most of the edible varieties grown in
the majority of the banana growing regions are susceptible to this
recently evolved most virulent strain, the TR4. TR4 destroyed the
Cavendish banana industry in Taiwan (Hwang and Ko, 2004), and
is now reported in Southeast Asia (Molina et al., 2009), China (Li
et al., 2013, 2014), Indonesia (Hermanto et al., 2009) and Northern
Australia (Ploetz, 2006). Also infestation was recently suspected
in Jordan (Ploetz et al., 2015), Pakistan, Lebanon (Ordonez et al.,
2015, 2016), Mozambique (Butler, 2013), Oman (Dita and Martínez-
de la Parte, 2014), and Northern Queensland, Australia (Ordonez
et al., 2015; ABGC, 2015) which are recognized as important banana
growing regions. Development of new banana cultivars through
genetic engineering with durable disease resistance will have sig-
nificant impact in managing this threatening disease (Table 2).
Over-expression of antimicrobial peptides belonging to plant or
animal origin have provided considerable resistance in trans-
genic banana plants (Chakrabarti et al., 2003; Ghag et al., 2012,
2014b; Mohandas et al., 2013). Most potent floral defensins when
ectopically expressed in transgenic banana plants were capable of
resisting the pathogen (Ghag et al., 2012). Although these defense
peptides imparted resistance, they may possibly also target ben-
eficial fungi associated with the banana roots. Considering this,
we have applied the RNA interference technology to generate Foc
specific siRNA in transgenic banana plants capable of targeting cog-
nate Foc transcript thereby preventing its invasion (Ghag et al.,
2014a). Boosting the plant defense response by over-expressing
proteins involved in unfolded protein response, chaperone activ-
ity and secondary metabolites is another approach which have
imparted durable resistance to this pathogen (Vishnevetsky et al.,
2011; Ghag et al., 2014c). These reports demonstrate the potential-
ity of genetic engineering technology in developing transgenic or
cisgenic banana plants resistant to Foc. Although the above studies
are confined up to greenhouse, field trials are necessary to affirma-
tively establish its importance under natural conditions.
Another fungal disease which has immense impact on the
banana production world over is the Sigatoka disease, also
known as leaf streak or spot disease. Black Sigatoka caused
by Mycosphaerella fijiensis and yellow Sigatoka caused by
Mycosphaerella musicola results in reduced green canopy, prema-
ture ripening and fruit defects. Chemical pesticides single or in
combination are often used to control this disease. However the
tendencies of the pathogen to develop resistance to these chemi-
cals and harmful effects on the environmental and banana workers
have become a great concern (Ma and Michailides, 2005; Avenot
and Michailides, 2010; Wesseling et al., 2001). Kovács et al. (2013)
expressed rice chitinases in transgenic banana plants and showed
that the necrotic leaf area was decreased by 73–94% compared to
the untransformed susceptible controls. As strongly speculated that
Mycosphaerella has a very short biotrophic phase of lifestyle dur-
ing infection (Friesen, 2016), an RNA interference strategy can be
designed to develop resistance to Sigatoka disease by construct-
ing ihpRNA vector using Mycosphaerella gene sequence, analogous
to the study carried out by our group to develop Fusarium wilt
resistance described in the preceding paragraph.
Banana bacterial wilt (BBW) caused by Xanthomonas campestris
pv. musacearum is predominant in the chief banana growing
regions in Africa where banana is a staple food and the disease poses
one of the greatest threats to banana production. Transgenic banana
plants expressing plant ferredoxin-like amphipathic protein (PFLP)
or hypersensitive response-assisting protein (HRAP) from sweet
pepper demonstrated resistance to BBW infection (Tripathi et al.,
2014b). The International Institute of Tropical Agriculture (IITA)
and the National Agricultural Research Organization (NARO) of
Uganda, along with the African Agricultural Technology Foundation
(AATF) carried out confined field trials of these transgenic plants
and showed that 20% of the hrap expressing lines and 16% of the
pflp expressing lines were fully resistant and retained the resistance
in the ratoon crop (Tripathi et al., 2014c).
Viral diseases caused by banana bunchy top virus (BBTV),
banana streak virus (BSV) and banana bract mosaic virus (BBMV)
cause significant yield reduction. Genes encoding for antiviral pro-
teins, antiviral systemic signal inducers, and viral genome specific
ribozymes have been engineered to confer viral resistance in other
plants (Gadani et al., 1990; Mandadi and Scholthof, 2013). Con-
versely, RNA interference has become the best technology to impart
antiviral defense in plants (Agius et al., 2012; Ibrahim and Aragão,
2015). Transgenic plants capable for producing viral specific dsRNA
which are processed into small interfering RNA, cleaves off the cog-
nate viral transcripts (such as replicase, coat protein, movement
protein) during infection thereby preventing replication and pro-
liferation of viruses. RNAi constructs designed to silence the BBTV
replicase gene were completely resistant to BBTV infection under
glasshouse conditions (Cheah et al., 2009; Shekhawat et al., 2012;
Hu et al., 2014).
Nematode infestation in the banana fields is a serious threat
to productivity. Burrowing nematode, Radophilus similis root-knot
nematode Meloidogyne spp. and Helicotylenchus multicinctus are
commonly associated with banana roots. These diseases results in
the destruction of root tissue, leaving plants with little to no sup-
port and decreases the capacity of water uptake and translocate
nutrients. Presently, nematicides are used as a control measure but
these are strong and harmful chemicals. In this regard transgenic
approach can be utilized as an effective strategy to boost plants
defense system or produce toxic proteins or specific proteinase
inhibitors capable of killing these nematodes. Nevertheless, host
induced gene silencing can be successfully applied to deliver resis-
tance by targeting nematode specific genes required for growth
and development (Fairbairn et al., 2007). The success of trans-
 S.B. Ghag, T.R. Ganapathi / Scientia Horticulturae 214 (2017) 91–98 95

Table 2
List of genes used to develop resistance in transgenic banana plants.

Target gene Transgenic plant/Cultivar Source of gene Target pathogen References

Fungal
Magainin Musa spp.cv. Rasthali Frog skin Foc race 1 Chakrabarti et al. (2003)
HL M. spp. cv. Taijiao Human Foc race 4 Pei et al. (2005)
Endo ˇ-1,3-glucanase gene M. spp.cv. Rasthali Soybean Foc race 1 Maziah et al. (2007)
pflp M. acuminata cv. Pei Chiao M. Sweet Pepper Foc race 4 Yip et al. (2011)
acuminate cv. Gros Michel
Bcl-xL, Ced-9 and Bcl-2 M. spp. cv. Lady Finger Animal Foc race 1 Paul et al. (2011)
3 UTR
tlp M. spp. cv. Nangka Rice Foc race 4 Mahdavi et al. (2012)
PhDef1, PhDef2 M. spp. cv. Rasthali Petunia flowers Foc race 1 Ghag et al. (2012)
chit42 M. spp. cv. Furenzhi Trichoderma harzianum Foc race 4 Hu et al. (2013)
Ace-AMP1 M. spp. cv. Rasthali Onion seed Foc race 1 Mohandas et al. (2013)
VEL and FTF1 M. spp. cv. Rasthali Fusarium oxyporum f. sp. Foc race 1 Ghag et al. (2014a)
cubense race 1
Sm-AMP-D1 M. spp. cv. Rasthali Chickweed Foc race 1 Ghag et al. (2014b)
MusaDAD1, MusaBAG1 and M. spp. cv. Rasthali Musa spp. cv. Rasthali Foc race 1 Ghag et al. (2014c)
MusaBI1
ThEn-42 and StSy M. spp. cv. Grand Nain Trichoderma harzianum and Mycosphaerella fijiensis Vishnevetsky et al. (2011)
grape
rcc2, rcg3 M. spp. cv. Gros Michel Rice Mycosphaerella fijiensis Kovács et al. (2013)
Bacterial
Pflp, hrap M. spp. cv. Sukali Ndizi and M. Sweet pepper X. campestris pv. musacearum Tripathi et al. (2014a)
spp. cv. Nakinyika
Xa21 M. spp. cv. Gonja manjaya Rice X. campestris pv. musacearum Tripathi et al. (2014b)
Viral
Rep M. spp. cv. Dwarf Brazilian BBTV BBTV Cheah et al. (2009)
BBTV-G- cp M. spp. cv. Williams BBTV BBTV Ismail et al. (2011)
Rep, ProRep M. spp. cv. Rasthali BBTV BBTV Shekhawat et al. (2012)
Nematode
Cystatin M. spp. Rice Radopholus similis Atkinson et al. (2004)
Cystatin, synthetic peptide M. spp. cv. Gonja manjaya Maize R. similis, Helicotylenchus Roderick et al. (2012)
multicinctus and Meloidogyne
spp

Foc—Fusarium oxysporum f. sp. cubense, BBTV—Banana bunchy top virus.

genic approach for defense against banana nematodes has also been
demonstrated by using maize or rice cystatin (Atkinson et al., 2004;
Roderick et al., 2012).
Banana weevil (Cosmopolites sordidus) also pose threat to banana
production as they damage root system thereby reducing nutrient
supply, reduced plant vigor and increased susceptibility to other
pest and diseases (Gold et al., 1999). Options for biological control
are restricted to use of pheromone traps, botanicals and cultural
practices which are not satisfactorily effective. Till now there are
no transgenic banana lines generated to resist weevil infestation
however in vitro studies have demonstrated the importance of pro-
tease inhibitors (Montesdeoca et al., 2005; Kiggundu et al., 2010)
and ubiquitin E2 gene (Ocimati et al., 2014) in managing banana
weevil.
3.3. Biofortified GM bananas
Deficiencies of minerals and vitamins affect billions of peo-
ple worldwide, with a sizeable number in developing countries
(Kennedy et al., 2003; Mayer et al., 2008). Deficiencies occurring
due to iron, zinc, iodine and vitamin A are tackled through sup-
plementation and food fortification (Dairo and Ige, 2009). One of
the best ways to supply micronutrients to the poor and needy in
developing nations is to make their staple food more nutritious
by either breeding or genetic engineering. Vitamin A deficiency
(VAD) is a public health problem identified in 118 countries, with
predominance in Southeast Asia and Africa, where diet includes
mostly cereals and tubers (Müller and Krawinkel, 2005). The team
at Queensland University of Technology (QUT), Australia has engi-
neered Cavendish banana with increased amount of ␤-carotene,
a precursor of vitamin A. The primary objective of this project
supported by the Bill and Melinda Gates foundation is to allevi-
ate the problem of infant mortality and blindness occurring due to
VAD in East Africa. The phytoene synthase gene (APsy2a) involved
in the biosynthesis of provitamin A was isolated from a variety
rich in ␤-carotene (Fe’i cv. Asupina) and introduced into Cavendish
banana which showed accumulation of 55 ␮g/g of dry weight ␤-
carotene equivalent in one of the field grown transgenic banana
lines (Mlalazi et al., 2012; Paul et al., 2016). Moreover, the same
group at QUT is working towards increasing micronutrient (such
as iron) content in bananas. Since iron deficiency assumes as yet
another facet of malnutrition, ferritin gene from banana is overex-
pressed in commercial cultivars to accumulate iron in the fruits.
Our research efforts in this direction has also exploited the use
of a soybean ferritin gene to increase iron content in transgenic
banana plants (Kumar et al., 2011). Further research is underway
in many institutions such as QUT (Brisbane, Australia), BARC (Mum-
bai, India) and NRCB (Trichy, India) to identify and stack genes
involved in the absorption, trafficking and accumulation of iron in
banana fruits.
4. Present challenges and future prospects
A major challenge to agricultural biotechnology is to increase
productivity with the available natural resources. The situation is
getting worsened due the dwindling of available water resources
for agriculture and the rate at which cultivable lands turning barren.
Further, it is also becoming difficult for the nations to feed an ever-
increasing population. The future nutrition needs cannot be fulfilled
by the current production rates. At present more than a billion
people are starving and several thousands are malnourished. The
principal objective is to develop food production, primarily of the
staple ones by proficient use of best available and productive tech-
nology. Introduction of appropriate traits for biotic resistance and
96 S.B. Ghag, T.R. Ganapathi / Scientia Horticulturae 214 (2017) 91–98
abiotic tolerance into banana plants will decrease losses occurring
due to pest and diseases and also increase the area under cultivation
thereby effectively contributing to increased food security. Genetic
transformation is considered to be the best strategy for banana
improvement because resistance sources are insufficient among
the genetic reserves available. Since most of the cultivated and
commercially important varieties are triploids and fruit develop
parthenocarpically, the foreign gene introduced may not transfer
to any other plant thereby reducing the environmental risks of gene
contamination. Plant biotechnology is a key player for banana crop
improvement in terms of pest and disease resistance and nutri-
tional quality improvement. GM bananas are more likely to address
the problem of chronic starvation and reduce reliance on the use
of polluting chemical pesticides. It is more importance to address
the food crisis issue than to politicize GM crops. Recently, Nobel
laureate Sir Richard John Roberts rightly said in his talk at the Uni-
versity of Mysore (Mysuru, India) that “To help people in need, we
need more science in politics and less politics in science” (Dec 15,
2015, Deccan Herald). Moreover, lack of public awareness, negative
opinion in the media, opposition by activist groups and political or
business mindsets of select few have always opposed GM crops. It’s
the responsibility of the government and regulatory authorities to
streamline and implement course of action in the areas of biosafety,
regulation and dissemination of GM food to public. Banana being
a staple food of developing nations and largest consumed fruit in
developed countries, transgenic bananas could be instrumental in
tackling malnutrition, poverty alleviation, food security and sus-
tainability.
Conflict of interest
Authors declare that no conflict of interest exists.
Acknowledgements
Authors would like to thank Head, Nuclear Agriculture &
Biotechnology Division, BARC for his support and encouragement
and Dr. P. Suprasanna for critically reading the manuscript.
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