Scientia Horticulturae 164 (2013) 578–588

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

Review

Molecular breeding to improve guava (Psidium guajava L.): Current

status and future prospective
S. Nimisha a , D. Kherwar a , K.M. Ajay b , B. Singh c , K. Usha a,∗
a
Division of Fruits and Horticultural Technology, Indian Agricultural Research Institute, New Delhi 110012, India
b
National Research Centre on Plant Biotechnology, India
c
Centre for Environment Science and Climate Resilient Agriculture, Indian Agricultural Research Institute, New Delhi, India

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

Article history: Guava (Psidium guajava L.) is often referred to as the apple of the tropics. It is a native of tropical Amer-
Received 3 August 2013 ica and has been naturalized in India. Being very hardy, it gives an assured crop even with very little
Received in revised form 10 October 2013 care. The main objectives of guava breeding are aimed at improving both plant and fruit characteris-
Accepted 16 October 2013
tics such as to develop high yielding, high quality dwarf varieties with fruits of uniform shape, good
size, attractive skin and pulp colour, fewer seeds and or soft seeds, resistant to wilt, long storage life,
Keywords:
suitable for table and processing purposes and to evolve wilt resistant and dwarfing rootstocks. Conven-
Guava
tional breeding has helped to a limited extent and it is high time that biotechnological tools are explored
Psidium guajava
Molecular marker
and exploited either alone or in combination with conventional breeding to improve the crop produc-
Breeding tivity and to address challenge of improving fruit quality, and tolerance to a biotic and biotic stresses.
Genomics Success of molecular breeding however, depends largely on available genomic resources which could
be exploited for marker aided selection (MAS) and for genetic transformation of non food traits. Guava
genomic resources are however scarce and inhibit researchers from exploiting biotechnology tools for the
development of improved guava varieties. An effort has been made in this paper to collate and critically
analyse the status of genomic advances in guava and their potential application for improving quality
and productivity of this important fruit crop.
© 2013 Elsevier B.V. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 578
2. Conventional methods for guava improvement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 579
2.1. Selective breeding . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 579
2.2. Heritability pattern and hybrid breeding . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 579
2.3. Polyploidy in guava . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 580
2.4. Molecular approaches for guava improvement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 581
3. Molecular markers for genetic diversity and germplasm discrimination . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 581
4. Molecular markers for fruit/flesh colour . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 582
5. Genetic transformation studies in guava . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 583
6. Genetic mapping . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 583
7. Current status of guava genomic resources . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 584
8. Prospective . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 584
9. Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 585
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 585

1. Introduction

Guava (Psidium guajava L.) is the most valuable cultivated

species of the Myrtaceae family popularly known as “poor man’s
∗ Corresponding author. fruit” or “apple of tropics” (Nakasone and Paull, 1998). It is native
E-mail address: kalidindi.usha3@gmail.com (K. Usha). to tropical America and is presently found distributed in many

0304-4238/$ – see front matter © 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.scienta.2013.10.017
 S. Nimisha et al. / Scientia Horticulturae 164 (2013) 578–588 579

tropical and subtropical countries (Cobley, 1976; Samson, 1986; Boonprakob, 2005). The floral structure (epigynous flower, with
Morton, 1987). Guava fruit is commercially important in India, abundant incurred stamens of various sizes), pollen viability, long
China, Indonesia, South Africa, Florida, Hawaii, Egypt, Yemen, juvenile period, self-incompatibility, and heterozygous nature limit
Brazil, Mexico, Colombia, West Indies, Cuba, Venezuela, New the scope of conventional breeding methods in guava (Jaiswal and
Zealand, Philippines, Vietnam and Thailand (Wilson, 1980; Yadava, Amin, 1992; Coser et al., 2012; Usman et al., 2013). Molecular tech-
1996; Le et al., 1998; Tate, 2000) and is popular due to its year niques and other emerging biotechnological tools are very helpful
round availability, rich nutritional and medicinal value, and afford- to study the extent of genetic variation among cultivars as well
able price, suitability for transportation, handling and consumer as to find out genetic markers associate with the specific trait like
preference. All the cultivars of Indian guava belong to a single pulp colour, soft seed, wilt resistance etc. This review highlights the
species P. guajava. However, several Psidium species like P. cat- major biotechnological advances made in guava breeding during
tleianum, P. molle, P. acutangulam, P. araca, P. chinensis, P. cujavillis, the past years.
and P. pumilum are available globally and at various research sta-
tions in India. There are more than 160 guava cultivars in India with
2. Conventional methods for guava improvement
huge demand in National and International market. Brazil, Mexico,
Thailand, Peru, Philippines are the major exporting countries of
2.1. Selective breeding
guava fruits and processed products (Varmudy, 2013).
Guava is often marketed as “super fruit”, being rich in vitamins
Guava is an open pollinated crop and seedlings were exten-
‘A’ and ‘C’ with seeds that are rich in omega-3, omega-6 polyun-
sively used to raise new plantations (Hayes, 1957). Hence guava
saturated fatty acids and especially dietary fibre. A single Apple
exhibits high levels of genetic diversity due to continuous cul-
Guava fruit contains over four times the amount of vitamin C as
tivation of heterogeneous seedling trees. Selection from these
that of a single orange i.e. over 200 mg per 100 g serving and also
seedlings can be used to obtain superior strains with respect to
has good level of the dietary minerals, potassium, magnesium, and
fruit yield and quality. Several attempts have been made to select
generally a broad, low-calorie profile of essential nutrients. Guava
superior types from seedling population (Singh, 1959; Phadnis,
contains both carotenoids and polyphenols – the major classes of
1970; Thonte and Chakrawar, 1982; Pathak and Dwivedi, 1988;
antioxidant pigments, giving them relatively high dietary antiox-
Iyer and Subramanyam, 1987; Rajan et al., 1996). Ruehl (1946)
idant value among plant foods. As these pigments produce the
observed variation in guava seedlings at the Subtropical Exper-
fruits’ colour, guavas that are red or orange in colour have more
iment Station, Homestead, Florida, USA. His work resulted in
potential value as antioxidant sources than yellowish-green ones
selection of 3 varieties, namely, Supreme, Red Indian and Ruby. One
(Singh, 2005). Despite the economic and health advantages, there
strain from open-pollinated seedlings of Allahabad Safeda collected
are a number of problems that impact guava productivity and qual-
from Lucknow, India was selected and released as Lucknow-49,
ity. Guava wilt, anthracnose, canker and several root pathogens
which became very popular for its spreading growth habit, glo-
cause severe losses (Mishra, 2005). An additional difficulty in guava
bose and large fruits having white flesh, high T.S.S. (12%), sugar:acid
breeding is the variation in fruit characteristics from one year to
ratio and yield (Cheema et al., 1954). Allahabad Surkha is a seedling
another, caused by environmental factors (Chezhiyan, 1988). There
selection from Allahabad, bearing large, uniformly pink fruits with
is also evidence of variation between fruits on the same tree and,
deep pink flesh (Nand et al., 1991). In Brazil, Canizares (1981) stud-
to a lesser extent, among trees of the same genotype (Thaipong
ied over 3000 seedlings by open-pollination and obtained about
and Boonprakob, 2005). This forces the breeder to test the same
60 promising clones. Five plants had fruits weighing more than
cultivars for many years before final characterization and use in
400 g each. Selection of superior seedlings resulted in the develop-
breeding programs.
ment of a number of varieties such as ‘Paluma’, the main Brazilian
The major problems facing the guava industry in different parts
variety and many others, like ‘Kumagai’, ‘Pedro Sato’, ‘Cortibel’.
of the world are severity of wilt disease, high seed content of diploid
In South Africa, Rensburg and Preez (1985) made 6 Nelspriut
commercial varieties and poor yield with small, irregular and mis-
guava selections and compared them with the standard cv. Fan
shapen fruits of triploid seedless varieties (Negi and Rajan, 2007).
Retief. Fruit weight was highest in selection No. 6 and ascorbic
The source of resistance to guava wilt was reported by Edward
acid content in selection No. 1. At the Indian Institute of Horticul-
and Shankar (1964), but research progress is slow. Hence devel-
tural Research (IIHR), Bangalore, India from 200 open-pollinated
opment of a wilt resistant, high yielding guava cultivar deserves
seedlings of variety Allahabad Safeda collected from Uttar Pradesh,
the first priority in guava breeding. Priority is also given to good
India a seedling selection, Selection-8 was found to be promising,
fruit quality because there is little merit in increasing yield and dis-
which was released as Arka Mridula. Plants of the selection are
ease resistance if not accompanied by quality. The need for longer
dwarf and give higher yield. The fruits are of medium size with
shelf life with improved texture has been felt for long. Other qual-
white pulp, few soft seeds, excellent quality in terms of sugar con-
ity parameters that need consideration are: (a) appearance which
tent and TSS and good shelf-life (Iyer and Subramanyam, 1988).
concerns uniform size and shape of fruits, attractive skin colour,
At Govind Ballabh Pant University of Agriculture and Technology,
(b) edible quality like flavour, fewer and soft seeds and pulp tex-
Pantnagar (Uttarakhand), India, a promising selection Pant Prab-
ture, (c) processing quality like pulp colour and texture, high pectin
hat was released. It is heavy yielder with small soft seeded round
and vitamin C content, and (d) resistant to insect pest like fruit
fruits. At the Central Institute for Subtropical Horticulture (CISH),
fly. Tree shape and size are important secondary characters that
Lucknow, India, 631 open-pollinated seedlings of the red coloured
are required to be combined in a new improved guava cultivar.
guava were evaluated for various characteristics and guava variety
Good branch angles, producing a spreading tree, are important
Lalit was released for commercial cultivation (CISH, 1999). During
to minimise the branch breakage when carrying heavy crops and
last 10–15 years guava improvement work from different parts of
to reduce the labour input on tree shaping, particularly in high
the world resulted in release of more than 150 superior selections.
density planting system. Precocity is a major yield component
and the breeding programme should aim to select early bearing
cultivars with dwarf or semi-dwarf habit and a high fruit: shoot 2.2. Heritability pattern and hybrid breeding
ratio (Morton, 1987; Ray, 2002; Dinesh and Iyer, 2005). Knowledge
on genetic variability and environmental effect help in designing Heritability of a trait, as a proportion of the phenotypic variation
crosses, evaluation methods and breeding strategies (Thaipong and that is attributed to genetic causes, has been a prime indicator and
580 S. Nimisha et al. / Scientia Horticulturae 164 (2013) 578–588
helpful in taking decisions for genetic improvement of economic
traits (Narain, 2010). Prediction of response to artificial selection
and the existence of wide genetic variability could be exploited for
developing guava hybrids through hand pollination (Zipori et al.,
2007). Preliminary observations showed that if there is no self-
incompatibility, which sometimes occurs (Chezhiyan, 1988; Singla
and Dhaliwal, 2003), fruit-setting can take place before anthesis,
as the stigma is already receptive (Chezhiyan, 1988; Nalini et al.,
1973; Pereira and Sao Jose, 1987). Rajan et al. (2005) reported vari-
etal cross incompatibility since neither fruit nor seed set could
be obtained. Germplasm conservation and guava improvement
programs through controlled artificial pollination are in practice
at India, Cuba, Colombia, Venezuela, USA, Malaysia and Nigeria.
Considerable research efforts have been made to estimate the her-
itability pattern in guava. It has been observed that commercially
important traits such as yield, fruit size, certain types of disease
resistance and quality characteristics (vitamin C, acidity, pectin
content etc.) are often low in the heritability category. None of
these characters are determined solely by major genes, although
basic genes, subject to the modifying effects of polygene, have
been identified for some quality characters such as skin colour and
acidity (Mohan Jain and Priyadarshan, 2009). Red pulp colour is
dominant to white pulp colour and is governed monogenic ally
(Soubihe and Gurgel, 1962; Subramanyam and Iyer, 1982; Rajan
et al., 2005). Dinesh and Vasugi (2010) studied inheritance pattern
using ‘Purple guava’ and ‘Arka Mridula’ as parents and observed
that hybrids segregate in a ratio of 3:1 for green leaf types and pur-
ple leaf types. It was also observed that the attractive pulp colour
and high yields of ‘Beaumont’ can be transferred to other white
sweet cultivars. Many cultivated red fleshed varieties were found
to be heterozygous for this character. A linkage was found between
red flesh colour and bold seed size (Rajan et al., 2005). Similarly
bold seeds were found to be dominant over soft seeds and this
character was also found to be governed monogenic ally. Obovoid
shape of fruit is dominant over round (oblate) and pyriform. Dinesh
and Yadav (1998) studied the F1 progenies of four crosses among
‘Apple Colour’ and three other guava varieties. They found that
genotypic variance was less than phenotypic variance for all the five
characters analysed (fruit weight, length, volume, width and TSS).
The coefficient of variation also followed the same trend, imply-
ing greater manifestation of these characters. The low genotypic
coefficient of variation indicated low degree of genetic variability
present in half-sib progenies. The higher phenotypic coefficients
of variation imply the greater manifestation of these characters.
The coefficients of variation indicated only the variability in dif-
ferent characters and did not indicate the heritable portion. The
heritability in narrow sense was observed to be moderately high in
fruit length (44.45%) and TSS (42.88%). Heritability was least in fruit
width (31.68%). Thus selection can be practiced to improve the yield
characters since these traits are controlled by additive effects. The
fruit weight had positive correlation with fruit volume, fruit length
and width. However, negative correlation was observed between
TSS and other four characters. The genotypic correlation was higher
than phenotypic correlation for all the characters except TSS. This
can be attributed to the relative stability of the genotypes. This hap-
pens not only when genes governing the traits are similar but when
environmental factors pertaining to it also have similar effects. Co-
heritability estimates were moderately high for most of the pairs
of characters. The TSS goes down with the selection of big sized
fruits. However, selection of medium sized fruits would not bring
down the TSS. Most economically significant fruit traits show quan-
titative variation which is controlled by a combination of genetic
and environmental factors. The portion of genetic and environ-
mental variances for fruit weight (FW), flesh thickness (FLT), flesh
weight (FLW), fruit firmness (FF), seed cavity weight (SCW), total
soluble solids (TSS), titratable acidity (TA), juice acidity (pH) and
ascorbic acid (AA) were estimated with eight genotypes by
Thaipong and Boonprakob (2005). A high proportion of genotype
variance was found with FW, FLT, FLW, SCW and AA indicating that
genetic improvement for these traits through breeding and selec-
tion was achievable. In another study, the length of fruit showed
high values of heritability in narrow sense, while the other traits
presented low and medium values (Pérez et al., 2012). Despite
several problems in guava breeding a few successful reports are
available (Ribeiro and Pommer, 2004; Thaipong and Boonprakob,
2005; Dinesh and Iyer, 2005; Patel et al., 2007; Pommer and
Murakami, 2008; Rodriguez-Medina et al., 2010). Guava improve-
ment work resulted in the development of hybrids such as ‘Safed
Jam’, ‘Kohir Safeda’ and ‘Arka Amulya’ in India, ‘Século XXI’ in Brazil,
was developed through inter varietal hybridization. Inter specific
hybridization between Psidium molle and P. guajava has yielded
hybrids which are resistant to wilt and are grafting compatible.
Although diploid commercial cultivars of guava have resulted in
production of quality fruit, yet high seed content restricts its con-
sumption and spread worldwide (Fatta Del Bosco et al., 1992).
Thus developing fruits with few and soft seeds have attracted the
attention of breeders and researchers globally and is an important
priority thrust area of the guava breeding program (Raza et al.,
2003; Negi and Rajan, 2007; Dinesh and Reddy, 2001; Dinesh and
Iyer, 2005; Rajan et al., 2005).
2.3. Polyploidy in guava
Seedless type is an important commercial attribute and ‘seeded’
and ‘seedless’ guava types have been recognized. However, those
commercially recognized as ‘seedless’ are not completely seedless
and types ranging from ‘seedless’ to ‘less-seeded’ can be identified.
The seedless types are nevertheless found to be pollen fertile. Cyto-
logical examination of the ‘seeded’, ‘less-seeded’ or ‘seedless’ types
has shown that differences in chromosome number as well as gene
dissimilarities contribute to this feature. The ‘highly seeded’ and
‘less-seeded’ types are diploids while the ‘highly-seedless’ types
are triploids (Raman et al., 1969). Among polyploidy, triploids are
of utmost importance as they produce seedless fruits (Raza, 2001).
Breeders have been investigating methods that allow sexual recom-
bination that result in progeny with nearly seedless fruit. Basic
chromosome number of P. guajava is X = 11 (haploid). A natural
triploid with somatic chromosome number of 2n = 33 was reported
by Kumar and Ranade (1952). The same chromosome number was
reported by Raman et al. (1971) in a triploid seedless variety of P.
guajava. Chromosome set of seedless guava: ‘Indonesian Seedless’
and ‘Bangkok Apple’ is triploid (2n = 33). Another seedless cultivar,
‘Sa1ithong’, has chromosome number equal to 2n = 22 (Srisuwan,
2003). A seedless type at Poona, India, was found to be a triploid
with 33 chromosomes. Java’s Seedless Red Guava, a cross between
red Jakarta guavas x white Thai is triploid. Seedless varieties of
guava have been found to be auto triploids with 33 chromosomes
(Kumar and Ranade, 1952; Majumder and Singh, 1964; Naithani
and Srivastava, 1966; Iyer and Subramanyam, 1971). Tetraploidy
in guava has been induced with colchicines treatment (Janaki
Ammal, 1951; Ram Kumar, 1975). In order to evolve a variety
with less seeds and high yield, crosses were made between seed-
less triploid and seeded diploid variety Allahabad Safeda at the
Indian Agricultural Research Institute, New Delhi. Out of the 73
F1 hybrid seedlings raised, 26 were diploids, 9 trisomics (2n + 1),
5 double trisomics (2n + 1 + 1) and 14 tetrasomics (2n + 2). They
showed distinct variation in tree growth habit and leaf and fruit
characters. Three tetrasomic plants had dwarf habit and normal
shape and size of fruits with less number of seeds (Majumder
and Mukherjee, 1972a,b; Mukherjee, 1977). Mohammad (1974)
reported obtaining 30 trisomics, 2 double trisomics, 1 tetrasomic
and 2 higher aneuploids in progenies of open-pollinated triploid
 S. Nimisha et al. / Scientia Horticulturae 164 (2013) 578–588
and triploid × diploid crosses. Reduction in growth and the size
of leaf distinguished aneuploids from diploids. Aneuploids, par-
ticularly trisomics, had promising qualities and may be useful in
developing plants with reduced seediness and possibly in providing
dwarfing rootstocks. Sharma (1982) identified a promising dwarf
rootstock aneuploid No. 82 (named later as Srijan) through selec-
tion made out of 48 different aneuploid seedlings at IARI, New Delhi.
The aneuploid No. 82 is a tetrasomic and induced substantial dwarf-
ing of Allahabad Safeda in terms of plant height, plant spread and
tree volume (Sharma et al., 1992). A cross between diploid Allah-
abad Safeda and seedless triploid was named as Arka Amulya and
was released from IIHR, Bangalore, India. Fruits are medium in size
with white flesh, soft seeds, high TSS and with good keeping quality.
2.4. Molecular approaches for guava improvement
Guava breeding through conventional approaches is a long
term and cumbersome process that relies on the arbitrary
rearrangement of existing genes between two closely related
parent plants. The combination of morpho-agronomic characters
and DNA molecular markers constituted a novel tool of great
utility to characterize guava germplasm, estimate diversity level
and parentage relationship among accessions, and also to recom-
mend genotypes with conservation and breeding potential. Clonal
identifications are traditionally based on various morphological
characters; however, morphological characters may not be reli-
able attribute to discriminate between the closely related guava
genotypes (Chandra et al., 2005b). Being a cross-pollinated species,
substantial variability exists in seedling populations grown in dif-
ferent regions (Srivastava, 2005). High heterozygosis could be a
merit if stringent selection is made and selected plants could be
used as parents for future breeding programs. Most of the cultivars
grown on a commercial scale are seedling selections from the well-
known parent cultivars (Jaiswal and Amin, 1992). A close genetic
relationship among cultivars, somatic mutations, and changes due
to environmental alterations can create problems in correct iden-
tification of germplasm. Genetic engineering has now opened
up new possibilities by allowing the transfer of individual genes
within or across species and compliments the classical breeding
approaches. Molecular markers are the main working tools for
genomics surveys. With the development and availability of an
array of molecular markers and dense molecular genetics maps
in crop plants, marker assisted selection has become possible for
traits both governed by major genes as well as quantitative trait
loci (QTLs) (Babu et al., 2004). DNA markers are the predomi-
nant types of genetic markers for marker assisted breeding (MAB).
Each type of markers has advantages and disadvantages for spe-
cific purposes. In recent years, different molecular markers (RAPD,
RFLP, AFLP, SSRs, ISSR, and VNTRS) have been employed for the
investigation of cultivar origin and taxonomic relationship of sev-
eral plant species (Schlotterer, 2004; Schulman, 2007; Bernardo,
2008; Arif et al., 2011; Abdel-Mawgood, 2012). These molecular
techniques have been widely used to monitor differences in DNA
sequence in and among species. They also allow the creation of
new sources of genetic variation by introducing new and desirable
traits from wild varieties into elite lines. While restriction fragment
length polymorphism (RFLP) markers have been the basis for most
genetic work in crop plants, amplified fragment length polymor-
phism (AFLPs) and simple sequence repeats (SSRs) are currently the
most popular techniques used due to ease in detection and automa-
tion. The adoption of the new marker system, single-nucleotide
polymorphisms (SNPs), is now highly preferred, with the increas-
ing amount of sequence information, and the determination of gene
function due to genomic research. Relatively speaking, SSRs have
most of the desirable features and thus are the current marker of
choice for many crops. SNPs require more detailed knowledge of
581
the specific, single nucleotide DNA changes responsible for genetic
variation among individuals. However, more and more SNPs have
become available in many species, and thus they are also consid-
ered an important type for marker-assisted breeding (Ribaut et al.,
2010; Correia et al., 2011; Faria et al., 2011). Some of the impor-
tant achievements made in guava through molecular approaches
are presented in Table 1.
3. Molecular markers for genetic diversity and germplasm
discrimination
Molecular approaches are useful for characterizing the genetic
diversity at cultivars or species level, for identifying genes of com-
mercial interest and improvement through genetic transformation
technology (Morand et al., 2002; Zeid et al., 2003). Molecular mark-
ers have been used to answer questions related to the management
of genetic variation, identity, and relationship in breeding and pro-
duction populations. It can be used from any tissue at any time
during the plant growth, and thus expedite the process of variety
identification and breeding, and help in overcoming the limitations
of traditional methods (Azofeifa-Delgado, 2006). Markers have
been successfully used to estimate out crossing rates, study parent-
age, and outside pollen contamination in seed orchards, parameters
that have also been valuable to provide general guidelines for risk
assessment of gene flow from plantation to natural stands (Rao
et al., 2008; Barbour et al., 2010). Detection of genetic variation is
also important for micro propagation and in vitro germplasm con-
servation to eliminate undesirable somaclonal variations. Based on
the morphological characters it is not always possible to discrim-
inate between closely related guava genotypes although several
morphological markers like fruit colour, leaf shape and size but
they may not be useful to discriminate between the very closely
related guava genotypes. Keeping above in view the efficacy of
both molecular and morphological parameters were tested by Sax-
ena and co workers (2007) to discriminate between individuals in
half sib population of P. guajava consisting of 6 half-sib progeny
(CISH-G-1, G-2, G-3, G-4, G-5 and G-6), Allahabad Safeda and 2
Psidium species. They used PCR based Random Amplified Poly-
morphic DNA (RAPD) and directed amplification of mini satellite
DNA (DAMD) markers to study the genetic diversity and related-
ness among 22 guava accessions comprising commercial cultivars,
breeding lines, and unimproved cultivars. Similarity between pairs
of cultivars calculated by DAMD analysis were in the range of
0.22–0.95, with the maximum similarity (0.95) between CISH-G-
4 and CISH-G-5 and least similarity was between P. acutangula
and G-6 (0.22). The clustering revealed that most of the cultivars
originated from the Indo-Gangetic plains and grouped together
while cultivars which are of exotic formed separate group (Bajpai
et al., 2008). In guava, recently, a few reports have been made on
assessment of genetic diversity using RAPD markers (Dahiya et al.,
2002; Prakash et al., 2002; Chen et al., 2007; Pessanha et al., 2011).
Sanabria et al. (2006) characterized 53 accessions of P. guajava
by random amplified microsatellites which were classified into 4
groups by discriminate analysis. SSRs, also known as microsatellite
markers have been widely utilized in plant genomic studies, and are
reported to be more variable than RFLPs and RAPDs. Microsatel-
lite markers to study genetic diversity in guava were developed
using a genomic library enriched for (GA)n and (GT)n dinucleo-
tide repeats and 23 nuclear SSR loci were chosen to assess the
diversity in three guava species. All the SSR loci were found to be
polymorphic after screening for diversity in different cultivars, and
across taxa amplification tests showed potential transferability of
most SSR markers in three other Psidium species (Risterucci et al.,
2005). In single primer amplification reaction method (SPAR) both
RAPD and inter-simple sequence repeat (ISSR) primers were used to
582 S. Nimisha et al. / Scientia Horticulturae 164 (2013) 578–588

Table 1
Molecular marker approaches for characterization and improvement in guava.

Molecular marker Research area Reference

DAMD Efficacy of both molecular and morphological parameters was tested to discriminate between Saxena et al. (2007)
individuals in half sib population of Psidium guajava.
Studied the genetic diversity and relatedness among 22 guava accessions. Bajpai et al. (2008)

RAPD Determined the genetic relationship in 13 north Indian cultivars of guava. Dahiya et al. (2002)
Analysed molecular diversity of 41 different genotypes of guava. Prakash et al. (2002)
Determined the phylogenetic relationship in 18 cultivars of Taiwan Chen et al. (2007)
Examined the diversity among twenty-two Psidium guajava L. cultivars. Sharma et al. (2007)
Identified molecular markers associated with high quercetin accumulation in the leaves of guava. Feria-Romero et al. (2009)
Studied cluster analysis in P. guajava samples Ahmed et al. (2011)
Analysed the genetic diversity between 20 guava accessions. Pessanha et al. (2011)

AFLP Morphological and genetic diversity was analysed in 48 Mexican guava germplasm. Hernandez-Delgado et al. (2007)
Studied genetic diversity of Mexican guava germplasm. Sánchez-Teyer et al. (2010)
Genetic similarity among accessions of guava and Brazilian guava araçazeiros. Correa et al. (2011)

ISSR Developed molecular marker for pulp colour in guava. Nadgav (2010)
Assessed genetic variability among eleven Psidium sp. Mani et al. (2011)
Assessed the clonal fidelity of micro-propagated guava. Liu and Yang (2012)

SSR Constructed (GA)n and (GT)n microsatellite-enriched library. Risterucci et al. (2005)
Assessed genetic variability in the collection. Valdés-Infante et al. (2007)
Transferred guava-derived SSR primers to other Myrtaceae species. Briceno et al. (2010)
Characterized diversity of guava germplasm in the United States. Viji et al. (2010)
Developed the guava microsatellite (SSR) markers using the SAT software. Risterucci et al. (2010)
Studied genetic diversity of the natural population of P. guajava in Venezuela. Aranguren et al. (2010)
Characterized guava cultivars and generated barcodes. Marker mPg-CIR09 was found to be the most Kanupriya et al. (2011)
suitable primer combination for guava accession characterization.
Characterized 69 cultivars of guava for higher nutritional content. Santos et al. (2011)
Characterized the genetic diversity among 28 P. guajava genotypes. Coser et al. (2012)
Discriminated the wild guava tree Nogueira et al. (2012)
Studied the genetic distance among guava genotypes. Noia et al. (2012)
Studied the genetic homogeneity of guava plants derived from somatic embryogenesis. Rai et al. (2012)
Studied the genetic diversity of wild guava and commercial cultivars from southern Espírito Santo. José et al. (2012)
Evaluated the genetic diversity of 66 wild guavas of six localities in the south of the state of Espírito Angélica et al. (2012)
Santo and in Caparaó, Minas Gerais, Brazil, by morphological descriptors and microsatellites.
Transferred simple sequence repeat SSR markers developed in guava (Psidium guajava L.) to four Rai et al. (2013)
Myrtaceae species.

distinguish the genetic variability and some of the primers showed
100% polymorphism, while average polymorphism in both marker
systems was 77 and 81.6% respectively. Dendrograms revealed two
main clusters, separating the genus, Feijoa sellowiana and Psidium
sp. with a genetic distance of 0.86 (Mani et al., 2011). Valdés-Infante
et al. (2007) and Viji et al. (2010) utilized microsatellite markers for
guava accession identification and germplasm characterization and
allelic variation was observed in each of the cultivars irrespective
of geographic origin. Coser et al. (2012) characterized the genome
and genetic diversity among 28 P. guajava genotypes by morpho-
logical, karyotypical, nuclear 2C-value and use of SSR markers. From
the 26 SSR loci used, 70 alleles were identified, varying from one
to five per locus, with an average of 2.7 alleles per locus. Among
these SSR, 24 were polymorphic and thus used for assembly of
dissimilarity matrix. Moreover, genetic divergence among culti-
vated and plant nursery genotypes could be observed, which was
in accordance with their single origins. Nogueira et al. (2012) eval-
uated the genetic diversity of 66 wild guavas of six localities in the
south of the state of Espírito Santo and in Caparaó, Minas Gerais,
Brazil, by morphological descriptors and microsatellites. Genetic
diversity was observed between and within the localities regard-
less the type of trait studied, indicating that existing variability can
be exploited in guava breeding and in conservation programs of
the culture. Noia et al. (2012) studied the genetic distance among
guava genotypes (P. guajava L.) through plants collected at differ-
ent altitudes, by microsatellite markers. They found that the wild
genotypes are of potential use in guava breeding to increase the
options of genotypes grown commercially and demonstrated the
cross-genera transferability of 23 SSR primer pairs developed for
guava (P. guajava L.) to four new targets, two species of eucalyp-
tus (Eucalyptus citriodora, Eucalyptus camaldulensis), bottlebrush
(Callistemon lanceolatus) and clove (Syzygium aromaticum), belong-
ing to the family Myrtaceae and subfamily Myrtoideae. The high
level of cross-genera transferability of guava SSRs may be applica-
ble for the analysis of intra- and inter specific genetic diversity of
target species, especially in E. citriodora, C. lanceolatus and S. aro-
maticum, for which till date no information about the EST-derived
as well as the genomic SSR is available. Hernandez-Delgado et al.
(2007) and Sánchez-Teyer et al. (2010) studied the AFLP analysis of
genetic relationship among guava cultivars grown in different parts
of Mexico. The AFLP- and SSR-based dendrogram clusters analysed
guava accessions into 2 main groups with at least 5 different sub
clusters without specific separation based on the region of origin.
The results show that Mexican guava is diverse and genetic variabil-
ity could be used for conservation, management, and development
of new varieties. Correa et al. (2011) analysed 88 accessions, 64 of
guava and 24 of Brazilian guava, collected in ten Brazilian States,
adopting for the cluster dendrogram UPGMA, considering the simi-
larity matrix of Jaccard’s coefficient of 149 polymorphic AFLP bands
from 16 combinations of primers EcoRI and MseI. Two major groups
were identified: one formed by accessions of guava and other with
accession to Brazilian guava, including some accessions of guava.
4. Molecular markers for fruit/flesh colour
Apart from characterization and assessment of chemical diver-
sity, Feria-Romero et al. (2009) used RAPD amplification method
to identify molecular markers associated with high quercetin
accumulation in the leaves of guava trees, selected from four
different Mexican agronomic regions. Nadgav (2010) tried to asso-
ciate SPAR markers with the pink colour of guava and found four
primers (FP1/RP1, FP1/RP2, FP2/RP2 and FP3/RP1), out of nine,
 S. Nimisha et al. / Scientia Horticulturae 164 (2013) 578–588
yielding polymorphic bands which proved promising for further
detailed analysis. Sixty-nine Psidium accessions were characterized
to develop new cultivars that relate with high nutritional compo-
nents (Santos et al., 2011). Kanupriya et al. (2011) characterized
nine guava cultivars using 23 microsatellite markers and generated
the barcodes, derived from the allelic variation of the microsatel-
lite loci that clearly differentiated the cultivars under study in two
groups viz: pink flesh varieties and white flesh varieties. The poly-
morphic information contents of the markers ranged from 0.340 to
0.900 with a mean of 0.749.
5. Genetic transformation studies in guava
Genetic engineering has been considered a promising produc-
tion alternative since it shortens the breeding period. For this
approach to gain wide acceptance an efficient micro-propagation
and regeneration procedure to produce large numbers of rooted
plants from unique plants is a prerequisite (Liu and Yang, 2011).
Genetic transformation opens the opportunity for genetic manip-
ulation of plants at cellular level and provides the means for
modifying single horticultural traits without significantly altering
other aspects of the phenotype (Singh et al., 2004; Krishna and
Singh, 2007). The main target of gene transfer techniques is to
produce improved varieties through the incorporation of horticul-
tural important genes into existing cultivars (Singh et al., 2004).
Fruit trees are considered to be recalcitrant material for genetic
transformation studies and the main impediment for genetic trans-
formation is the regeneration of transformed plantlets. Choice of
explants having competence for transformation and regeneration
is a crucial factor. Hence, efficient tissue culture techniques become
the base for genetic transformation studies (Giri et al., 2004).
Genetic transformation studies aimed at improving morphological
and growth characteristics of the plants are however hindered by
low transformation efficiencies and genotype dependence of pro-
tocols. Guava regeneration studies are an essential complement
of transformation studies. The successful regeneration of geneti-
cally transformed plants has been achieved in several tropical fruit
plant species (Gomez-Lim and Litz, 2004; Giri et al., 2004; Singh
et al., 2004; Touchell et al., 2008). But genetic transformation sys-
tem for guava is still not well developed (Rai et al., 2010). An
engineered Agrobacterium tumefaciens strain LBA 4404 harbouring
binary vector pBI121 having selectable markers (nptII and GUS)
with CaMV 35S promoter gene has been used for transformation of
guava (Biswas et al., 2007). Preliminary work on genetic transfor-
mation of guava with cold hardiness genes (CBF1, CBF2 and CBF3)
was successfully demonstrated by Biswas et al. (2005, 2007). How-
ever, complete regeneration of transformed plants could not be
achieved. The development of recombinant DNA technology has
not only extremely impacted our understanding of gene struc-
tures, functions, and regulations, but also greatly facilitated gene
cloning, characterization, and their expression into target species.
Globally, attempts to engineer fruit crops are intended mainly to
incorporate traits which include delayed ripening, virus resistance
and improved processing and nutritional quality of the crops. For
instance, insertion of genes controlling ethylene biosynthesis could
be helpful in increasing shelf life of guava fruits. Transformation
of genes encoding hydrolytic enzymes such as chitinase and glu-
canase which can degrade fungal cell wall could also be beneficial in
development of wilt resistant plant of guava (Chandra et al., 2005b).
Guava fruit was identified as a particularly rich source of
hydroperoxide lyase (HPL) activity. The enzyme HPL is widely
distributed in plants and is involved in the biosynthesis of volatile
aldehydes and alcohols that are important constituents in the char-
acteristic flavour of fruits, vegetables, and leaves. HPL catalyses the
cleavage of 13- and 9-hydroperoxides of linoleic and linolenic acid
583
into volatile C6- or C9-aldehydes and C12- or C9-oxoacids, respec-
tively (Kim and Grosch, 1981). The C6 and C9 volatile compounds
have a commercial value in the production of natural flavour in
the food industry, and are potentially important in plant defence
against pathogens (Croft et al., 1993). The HPL enzyme purified
from guava fruits was cloned by polymerase chain reaction (PCR)
using 30 and 50 rapid amplification of cDNA ends (Tijet et al.,
2000). The sequence shows approximately 60–70% identity to
the known 13-hydroperoxide lyases. The guava enzyme shows a
high similarity to known HPL from other plant species. Prior work
with the guava HPL confirmed that the enzyme shares homology
with cytochrome P450, placing it in the cytochrome P45074B
subfamily as CYP74B5. One guava HPL gene was sub-cloned into
a plant transformation vector, and a transformation protocol was
performed in order to analyse the effects of HPL gene silencing
in the guava transformants (Valecillos and Fermin, 2008). Such
efforts will ultimately provide the most rapid advances in guava.
Transgenic trait modification, once successfully proven, should be
well adapted to the clonal propagation system used in industrial
Eucalyptus forests. Besides the potential economic impact of a
genotype independent transformation system, an efficient trans-
genic technology for Eucalyptus would represent a fundamental
step to advance functional genomics studies. To mitigate the
biological limitation for the study of wood formation using mutant
phenotypes, in vitro wood formation systems have been employed
to introduce transgenes transiently or stably into growing Eucalyp-
tus wood-producing tissue (Spokevicius et al., 2005) and recently
used it to show that ␤-tubulin determines cellulose microfib-
ril orientation during xylogenesis in E. globulus (Spokevicius
et al., 2007). The tree biotechnology company ArborGen recently
started work on genetically modified freeze tolerant Eucalyp-
tus trees to provide an economically viable hardwood option
(http://www.arborgen.us/uploads/press-releases/Dear BRS Stake
holder.pdf, accessed on 1/4/2011). Public institutions have usually
played a key research role for the Development of horticultural
crops, and this is also true of guava biotechnology. It is imperative
to increase research support in cases where there is a convincing
public interest, such as development of nutritionally enhanced
food products or when serious diseases threaten guava business
and a biotech-based answer is the most practical alternative for
developing resistant varieties. Unfortunately public institutions
generally have only limited access to the facilitating technolo-
gies and genes, making public–private partnerships scarce but a
striking avenue for development.
6. Genetic mapping
Modern technologies of gene sequencing, microarray experi-
ments, attempts made to understand gene and protein expression
within the cell of an organism and information on molecular
markers have been extremely helpful in identifying regions on
chromosomes (QTL) that bring about variation in a trait, thereby
providing tools that can lead to far more accurate selection pro-
cedures for genetic improvement (Narain, 2010). Advances in
genomics, the study of the DNA structure of genomes, has led
to complete genetic mapping of many fruit species increasing
the possibilities to locate useful genes affecting quantitative traits
(Soto-Cerda and Cloutier, 2012). The combination of advances in
informatics, sequencing technology, and genomics makes it the-
oretically possible to select promising types on the basis of the
genotype instead of the phenotype (Li and Homer, 2010; Treangen
and Salzberg, 2012). This has potential benefits to guava breeding
where the cost of phenotype selection are very high because of
the long time necessary for fruiting and the large amounts of land
required for each seedling. Genome-wide dissection of quantitative
584 S. Nimisha et al. / Scientia Horticulturae 164 (2013) 578–588
traits require the construction of complete genetic linkage maps
with sufficient genomic coverage and large segregating popula-
tions to allow detection of moderate to large effect quantitative trait
loci (QTLs). Some genomic resources have been generated within
the EU-Project “Improvement of guava: Linkage mapping and QTL
analysis as a basis for marker-assisted selection (GUAVAMAP;
FP6-2003-INCO-DEV-2 No. 015111)”. SSR markers for P. guajava
were developed massively. These were used to characterize guava
germplasm collections in different countries at the molecular
level. Genomic cosmid (cos sites + plasmid = cosmid) library was
established in guava (Ritter, 2012). Molecular hybridization was
done with heterologous probes for resistance (RGL sequences) and
homeotic genes (MADX-box and HOMEO-box genes). PCR primers
were designed for these sequences for downstream applications
(Grattapaglia et al., 2012). On the other hand, linkage maps were
constructed in different genetic backgrounds and QTL analyses
for several useful traits were conducted. Individual and combined
parental linkage maps have been constructed in three mapping
populations (Enana x N6, Enana x Suprema Roja and Enana x Belic L-
207) based on AFLP, SSR and Conserved Ortholog Set (COS) markers.
These population maps contain between 500 and 1000 markers and
have lengths of 1500–2200 cM each. Individual linkage groups vary
between 150 and 240 cM in length and contain between 35 and over
100 markers each (Rodríguez et al., 2010). Also an integrated guava
reference linkage map was established based on common mark-
ers map with a very high marker density of over 1700 markers.
In addition QTL analyses have been performed in these popula-
tions. The traits for QTL analysis include plant height, petiole length,
leaf length and width, yield, fruit number and average fruit weight,
fruit length and width, internal and external pulp thickness, seed
number and weight, vitamin C contents, acidity, and total soluble
solids. Over 100 QTLs were detected for the different characters
in the three populations and varied between 2 and 13 QTLs per
trait. Individual QTLs explained between 5 and over 40% of the
total variance. Total variance explained by the sum of all detected
QTLs varied between 20 and over 50% between traits and popula-
tions (Grattapaglia et al., 2012). Some of the QTLs are co-located
or closely linked to SSR markers, which allows an efficient selec-
tion for marker assisted breeding in different genetic background.
Several QTLs for the same or for related characters occurred at the
same location in different maps, allowing in this way to compare co-
locations of QTLs in different genetic backgrounds (Valdés-Infante
et al., 2003, 2012; Rodríguez et al., 2007).
7. Current status of guava genomic resources
Myrtaceae are strong candidates for genomic undertakings
that demand physical manipulation of DNA such as positional
cloning. Moreover, the upcoming availability of a reference genome
sequence for Euclayptus grandis, together with increasingly power-
ful high-throughput sequencing technologies and decreasing cost
will provide exceptional opportunities for whole genome com-
parative and evolutionary studies across genera in the Myrtaceae
(Grattapaglia et al., 2012). The use of advanced automated genome
sequencing technology like second generation sequencing (RNA
and ChIP Sequencing, Illumina, Solid, Roche (454) etc.) and most
recent third generation sequencing technology based on the sin-
gle molecule real time sequencing (SMRT), have helped to produce
thousands or millions of sequence reads concurrently in few hours
or days (Ronaghi et al., 1998; Shendure et al., 2005; Turcatti et al.,
2008; Eid et al., 2009). National Centre for Biotechnology Informa-
tion (NCBI) the world’s biggest database which contains all type
of genomic and proteomic sequence information from microbes
to virus, animals to plants has very little information on guava.
Genomic resources at NCBI are scarce and currently, for Psidium
gujava only 58 nucleotide, 22 GSS (Genomic Survey Sequences),
9 protein sequence, 13 EST (Expressed Sequenced Tags) and 86
CDS (coding DNA sequences) are available and a few more for
other Psidium species. The genome size of guava is ∼400 Mbp
which is distributed on 11 chromosomes (Feuillet et al., 2011).
Comparative availability of guava genomics with other major
fruit crops using NCBI database is given in Table 2. Few other
databases are available which contains only the information related
to medicinal use and general information is present like RAIN-
TREE (http://www.rain-tree.com/guava.htm#. Ud1Du6WaXbs),
guava-USDA Plants Database (http://plants.usda.gov/java/profile?
symbol=PSGU), Feedipedia (http://www.feedipedia.org/node/111)
etc. There is not a single database which contains the sequence
information on guava genomic or transcriptomic sequences. This
scarcity on the guava genomic sequence resources is inhibiting
researchers from exploiting the current biotechnology tools for the
development of improved guava varieties. The continuous decrease
in sequencing costs will stimulate the generation of new genomic
resources in the near future. In this context also the large num-
ber of genomic resources and particularly the upcoming genome
sequence of the related E. grandis will be useful based on poten-
tial synteny between related species (Ritter, 2012). It is highly
desirable that an emphasis is laid on developing robust, functional
and reproducible methods of genetic transformation in guava crop
to efficiently utilize the genomics information for identification
of key genes regulating productivity via the functional genomics
approach.
8. Prospective
Guava crop conventionally propagated through cutting, graft-
ing, stooling or air layering (Thorpe et al., 1991; Chandra et al.,
2004; Mitra and Sanyal, 2004) is limited by several pathogens
(Litz and Jaiswal, 1991). Few and soft seeds preferred by con-
sumers directs the need for aneuploidy breeding, development of
autotetraploids and in vitro genetic manipulation of somatic cells
(Pommer, 2012). Conventional breeding has many difficulties in
guava triploid production due to the reproductive complications
of the plant. To expedite genetic improvement, new approaches
need to be explored and exploited. Emerging bio techniques for
tissue culture and micro propagation of superior guava cultivars
have been reported by several researchers (Amin and Jaiswal, 1988;
Jaiswal and Amin, 1987; Loh and Rao, 1989; Papadatou et al., 1990).
A technique for successful in vitro propagation of guava germplasm
using shoot-tip explants from mature trees has been reported by
Jaiswal and Amin (1987). Rapid multiplications of seedling plants
of guava through in vitro shoot-tip culture and subsequent plant
establishment also have been successful (Papadatou et al., 1990).
Somaclonal variation, which normally occurs in many tissue cul-
tures (Larkin et al., 1985; Evans and Sharp, 1988; Lee and Phillips,
1988), could be useful for selecting guava plants resistant to wilt
disease. An efficient protocol for plant regeneration from callus
culture would also be helpful in selecting plants resistant to dis-
ease or environmental stresses. Organogenesis has been induced
in vitro both from mature tree explants (Amin and Jaiswal, 1987)
and seedling explants of guava (Singh et al., 2002; Shah et al., 2008).
Recovery of plants of haploid origin from anther/pollen culture
of guava could offer advantages in breeding (Jaiswal and Amin,
1992). The advances in biotechnological tools using cell, tissue,
and organ culture techniques have considerable potential for the
improvement of economically important trees within a limited
time frame (Giri et al., 2004; Singh et al., 2004). Considerable efforts
have been made for in vitro regeneration via somatic embryogen-
esis (Chandra et al., 2005a; Biswas et al., 2007; Rai et al., 2007).
Generation of new and promising variability through somaclonal
 S. Nimisha et al. / Scientia Horticulturae 164 (2013) 578–588 585

Table 2
Comparative availability of guava genomics to 6 other major fruit crops following NCBI database.

Genome information Guava Wine Grape Papaya Apple Mango Banana Sweet orange
(Psidium guajava) (Vitis vinifera) (Carica papaya) (Malus domestica) (Mangifera indica) (Musa x paradisiaca) (Citrus sinensis)

Nucleotide 86 160,132 4812 5100 673 38,630 1632

Nucleotide EST 0 446,668 77,393 325,020 1665 11,154 214,598
Nucleotide GSS 22 229,272 44,408 76 0 9 3
Protein 31 77,774 1624 2833 271 327 760
Structure 0 15 32 5 0 0 0
Genome 0 1 1 1 1 1 1
Popset 8 36 35 43 12 14 26
SNP 0 470,835 0 7907 0 3 0
GEO Datasets 0 1142 22 65 0 0 193
UniGene 0 22,258 6992 22,493 0 0 15,422
UniSTS 0 976 828 195 18 119 143
Gene 0 31,317 192 61 0 0 142

variant selection, production of androgenic, and gynogenic hap-
loids to achieve homozygous plants with stress-tolerance and free
from diseases through shoot tip culture need to be standardized
(Touchell et al., 2008). Rai et al. (2012) evaluated genetic homo-
geneity of 1-year-old guava (P. guajava L.) plants developed from
in vitro somatic embryogenesis with 6 SSR and 10 ISSR primers
and the monomorphic banding pattern in micro propagated plants.
The mother plant confirms the genetic homogeneity of the in vitro
raised plants and demonstrates the reliability of in vitro propaga-
tion system for guava. Twenty-one ISSR primers were employed to
test 30 progenies of clonal propagated guava plants. The similar-
ity indicators between the progenies and the mother ranged from
0.92 to 1.0. Such a similarity indicated a very low polymorphism.
The low somaclonal variation rate of 1.65% indicated the genetic
stability of micro-propagated guava. Therefore, mircopropagation
is a safe mode for multiplication of true to type plants of guava. This
is also very important for commercial propagation of elite clones
with desirable fruit traits (Liu and Yang, 2012).
9. Conclusion
Guava is nutritionally valuable crop with large potential in
domestic and export market. With current advances in biotech-
nology, strategies to improve the productivity and quality of guava
under challenges of abiotic and biotic stress are the need of the
hour. Genetic advances and tools like MAS could be highly use-
ful for parentage analysis, clonal fingerprinting, genetic diversity
analysis, development of genetic linkage maps, in disease and pest
resistance breeding and in relation to flavour, colour, fragrance,
vitamins, fruit softening, and other traits of interest. Availability of
genomic resource for Psidium and reliable genetic transformational
protocol is another constraint. This hinders our understanding of
variability among Psidium species, linkages, trait linked gene iden-
tification and transfer of genes within or across sources to improve
guava for commercial success. With the advent of robust sequenc-
ing techniques and research focus on developing transformation
efficient protocol for guava, the opportunities to increase the rate of
genetic gain in guava breeding programs and challenges of improv-
ing guava seems highly achievable. Marker assisted selection could
offer significant help but much genetic work must still be pursed
to locate these quality traits.
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