952

Review Article

APPLICATION OF TISSUE CULTURE TECHNIQUES IN POTATO

APLICAÇÃO DA CULTURA DE TECIDOS VEGETAIS EM BATATA

Tâmara Prado de MORAIS1; Simone Abreu ASMAR1; Herick Fernando de Jesus SILVA2;
José Magno Queiroz LUZ3; Berildo de MELO3
1. PhD in Plant Science, Instituto de Ciências Agrárias - ICIAG, Universidade Federal de Uberlândia - UFU, Uberlândia, MG, Brazil. 2.
Doctorate student in Plant Science, ICIAG - UFU. 3. Associate professor, ICIAG - UFU. jmagno@ufu.br

ABSTRACT: Potato is the world’s most important non-cereal food crop, and therefore, it is considered one of
the major food sources for humankind. Its conventional propagation is asexual, by using the tuber, which allows the
accumulation and dissemination of pathogens to new cultivation areas. This fact not only impairs the yield of this
solanaceous plant, but also threatens the maintenance of genotypes for commercial or breeding purposes. Due to the
impossibility of using botanical seed, conservation and exchange of germplasm of this species by means of conventional
methods are not feasible. In all potato-producing regions, the demand for high-quality tubers has been paramount to ensure
crops production. Thus, biotechnological techniques based on tissue culture are very important. Plant tissue culture offers
alternative methods of propagation by in vitro techniques that provide production and multiplication of material with high
sanity. Thus, this literature review summarizes the history and current situation of tissue culture techniques applied to
potato crop. Besides clonal multiplication, this biotechnological tool makes available initial indexed material to breeding
programs and certified seed potato, and facilitates the exchange and conservation of germplasm. For all these reasons, the
use of these techniques in potato production chain directly benefits producers by providing high-quality propagules.

KEYWORDS: Solanum tuberosum L. In vitro cultivation. Seed potato. Pathogen elimination.

INTRODUCTION exchange, provide the study of this species and of its

interactions with biotic and abiotic factors, and to
Potato (Solanum tuberosum L.) is an annual produce genetically modified plants and pathogen-
solanaceous plant from the Andes (MENEZES et free seed potatoes. This literature review
al., 1999). Its commercial product, the tuber, is the summarizes some relevant results obtained with
underground stem adapted to work as reserve source tissue culture in potatoes from 1977 to 2018. We
and reproduction mean (FORTES; PEREIRA, apologize in advance for the papers not included in
2003b). It is a culture of outstanding importance this review, due to the limited length of our
worldwide for having socio-economic manuscript, and encourage more publications on
repercussions, and for being one of the major food this subject to contribute to or innovate existing
sources for humankind. Potato is cultivated in about knowledge.
150 countries (BASERA et al., 2018), and is the
most important non-cereal food crop in the world TISSUE CULTURE IN POTATOES
(BAMBERG et al., 2016; NIKITIN et al., 2018).
Last harvest season, Brazil produced 4.3 million Micropropagation
tons of potato, in an area of 803,900 hectares In vitro establishment
(IBGE, 2017). Among plant tissue culture techniques,
On a commercial scale, potato has micropropagation has the greatest practical
vegetative propagation. However, this type of application (reviewed in SHAHZAD et al., 2017).
multiplication can accumulate several systemic Several studies have been reported in the literature,
fungi, bacteria and virus infections that provoke aiming at the optimization of potato
degeneration in the plants, culminating with yield micropropagation protocols (ÁVILA et al., 1994;
and vigor losses (NIKITIN et al., 2018). Thus, the PEREIRA; FORTES, 2000, 2004; DUTRA et al.,
use of virus-free propagative material, with high 2010; VENKATASALAM et al., 2013). However,
phytosanitary, physiological and genetic quality is one of the greatest difficulties in the development of
of great importance to ensure that the plant these protocols is the in vitro establishment of
expresses its maximum yield potential. plants, mainly due to fungal, viral and bacterial
In this scope, plant tissue culture can be contaminations.
applied to potato production chain in order to Fungi and yeasts can be controlled by
efficiently propagate the material of interest, eliminating the contaminated material (LEIFERT;
maintain germplasm banks, facilitate genetic WOODWARD, 1998), or by previously sanitizing

Received: 12/06/17 Biosci. J., Uberlândia, v. 34, n. 4, p. 952-969, July/Aug. 2018

Accepted: 20/12/18
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Application of tissue… MORAIS, T. P. et al.

the explants. In this case, potato shoots are Multiplication

disinfested by immersion in 70% alcohol for 15 Several parts of the mother plant can be
seconds, followed by immersion for 15 minutes in used as explant source at the multiplication stage. In
1.5% sodium hypochlorite solution with 0.1% vitro potato regeneration protocols have been
Tween-80, and rinsed three times with distilled and reported using immature zygotic embryos
autoclaved water (PEREIRA; MATTOS; FORTES, (PRETOVA; DEDICOVA, 1992), stem segments
2003). (GARCÍA; MARTÍNEZ, 1995; SANTIAGO et al.,
Viruses can be physically (heat) or 2012; KAUR; REDDY; KUMAR, 2017; ALI et al.,
chemically (by antiviral compounds) eliminated. In 2018; SILVA FILHO et al., 2018), leaves
a study developed by Nascimento et al. (2003), (CEARLEY; BOLYARD, 1997; KAUR; REDDY;
potato microcuttings with positive reaction to Potato KUMAR, 2017), tuber discs (ESNA-ASHARI;
virus Y (PVY) were subjected to thermotherapy VILLIERS, 1998), meristems (BADONI;
(37ºC) and chemotherapy [Ribavirin (RBV), 5- CHAUHAN, 2009), and shoot tips (DUTRA et al.,
Azacitidine (AZA) and 3-Deazauridine (DZD)]. The 2010).
thermotherapy, during 30 and 40 days, eliminated Due to the high susceptibility of potato to
20 and 37.5% of PVY, respectively. Among the diseases, especially to viruses, several studies have
chemotherapeutic agents, RBV presented the best started micropropagation by using meristems and
eradication rates, and obtained 55.5% of healthy shoot tips as explants in order to produce virus-free
plants. These authors also verified that simultaneous seed potato. However, some aspects may
treatments of thermotherapy and chemotherapy compromise the success of this technique, such as
showed higher efficiency in virus elimination, the delay in the differentiation of the meristematic
reaching percentages of 83.3, 70.0, and 50.0 of tissue in the culture medium, the possibility of
healthy plants, with RBV, AZA, and DZD, contamination (FORTES; PEREIRA, 2003a;
respectively. PEREIRA; MATTOS; FORTES, 2003), and explant
For the control of bacterial contamination, survival difficulty due to its small size.
the use of substances antagonistic to these The relationship between explant size,
microorganisms is required. The application of survival rate and pathogen elimination were
antibiotics in the decontamination process may be observed by El-Fiki et al. (1992). In this work,
beneficial since bacteria compete with plants for the seven potato cultivars were evaluated to determine
nutrients of the culture medium, and may excrete the ideal size of the shoot tip and the efficiency in
toxic substances to explants (SCHERWINSKI- PVX eradication (Potato virus X). Higher seedling
PEREIRA, 2010). Antibiotics may be incorporated survival and growth rates were obtained by using
into the culture medium or directly used on larger explants (0.4 mm), although they presented
contaminated explants (SALEHI; KHOSH-KHUI, smaller percentage of healthy plants (21.2) when
1997). In potato tissue culture, Pereira, Mattos and compared with the explants of 0.2 mm (47.4).
Fortes (2003) reported that the antibiotics Despite the proven efficiency of meristem
ampicillin, chloramphenicol, streptomycin and culture in pathogens eradication, the combination of
tetracycline in concentrations varying from 32 to this technique with antiviral treatments to ensure
256 mg L-1 inhibited the growth of endophytic pathogen elimination is often necessary. Wang et al.
bacteria. Besides acting on microorganisms, these (2006) reported higher frequency in the regeneration
antibiotics can also affect explants survival. The of PLRV-free (Potato leafroll virus) and PVY-free
addition of tetracycline and chloramphenicol to the (90 and 93%, respectively) potato seedlings by
culture medium adversely affects the in vitro applying thermotherapy, followed by meristem
development of potato explants; conversely, culture. The in vitro technique itself provided the
ampicillin has no phytotoxic effects on growth and regeneration of only 56% of healthy plants for
multiplication rate, and may be indicated for in vitro PLRV, and of 62% for PVY.
decontamination of this species (PEREIRA; The sanity of the micropropagated material
FORTES, 2003b). is guaranteed by indexing tests. The ELISA test
In vitro established plant material is more (Enzyme-Linked Immunosorbent Assay) has been
readily used as an explant donor than field-grown widely adopted for this purpose, since it is fast and
material (as surface disinfestation treatments are no allows detecting virus persistence in a large number
longer required), once the established material is of regenerated plants (NAIK; BUCKSETH, 2018).
carefully handled under aseptic conditions Molecular techniques based on the use of primers
(FORTES; PEREIRA, 2001; PEREIRA; FORTES, for specific regions of the virus genome, such as
2003a, b, 2004a; GONÇALVES et al., 2009). RT-PCR, are also largely used (LORENZEN et al.,

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Application of tissue… MORAIS, T. P. et al.

2006; 2008; MUÑOZ et al., 2018; NIKITIN et al., quantitative effect was observed (RAVNIKAR;
2018). In relation to the use of polyclonal GOGALA, 1990; RAVNIKAR; VILHAR;
antibodies, the technique involving RT-PCR has the GOGALA, 1992; ZHANG et al., 2006; KUMLAY,
advantage of differing strains and genetic variants, 2016).
distinguishing the types of viral isolates. The Salicylic acid, in concentrations from 30 to
indexation is of fundamental importance in the 120 mg L-1, has been used in association with
production chain of seed potatoes, as it ensures the mannitol (87.6 mM) for the in vitro preservation of
quality of the material obtained by means of potato (FORTES; PEREIRA, 2001). In addition,
meristem culture, reflecting in field yield, which can ethylene is associated with the formation of
be at least twice as high as the yield of plants hyperhydric structures (PARK et al., 2004), and the
derived from tubers (NAGIB et al., 2003). supplementation of the medium with ethylene
The chemical composition and physical synthesis inhibitors, such as silver thiosulfate, is
state of the culture medium may also interfere with often necessary (PERL; AVIV; GALUN, 1988;
potato in vitro multiplication. Regarding the MÖLLERS; ZHANGM; WENZEL, 1992;
chemical composition, the type of carbohydrate SARKAR; KAUSHIK; NAIK, 1999; ZOBAYED;
added to the culture medium affects explants ARMSTRONG; ARMSTRONG, 2001; ROSTAMI;
survival, as verified by Fortes and Pereira (2001). In EHSANPOUR, 2010). Similarly to jasmonic acid,
this work, higher survival rate was obtained when polyamines favor in vitro potato tuberization
the carbon source was sucrose in detriment to the (PROTACIO; FLORES, 1992; FERAY et al., 1993;
use of mannitol. Moreover, several studies mention MADER, 1995), while brassinosteroids have been
that 3% of sucrose is the ideal concentration for the used in the induction of embryogenic calli (MORE
optimal micropropagation of potato, which, in et al., 2001), acclimatization (AGRAMONTE et al.,
addition to promoting greater biomass 1996; PAVLOVNA et al., 2015), and proliferation
accumulation, favors the synthesis of sucrose, (BASERA et al., 2018). Although researchers have
glucose and fructose, which may play an important already taken the first steps for the study of these
role during potato plants acclimatization (WANG; regulators, there are gaps regarding the best
HU, 1982; DONNELLY; COLEMAN; COLEMAN, concentrations and combinations at different stages
2003; BADR; ANGERS; DESJARDINS, 2011; of potato micropropagation.
BANDINELLI et al., 2013). In relation to the consistency, the liquid
Although it has been reported that the use of culture medium is more efficient in potato in vitro
growth regulators in potato meristem culture is multiplication when compared with the semi-solid
dispensable (ROSELL et al., 1987; GOPAL; (PEREIRA; FORTES, 2003a). According to the
MINOCHA; DHALIWAL, 1998), the addition of same authors, higher biomass production is obtained
indole-butyric acid, indole-acetic acid, and when the material is cultivated in liquid medium
naphthalene-acetic acid, even at low concentrations, composed of MS salts at full concentration,
increases and accelerates the production of virus- supplemented with pantothenic acid (5.0 mg L-1),
free seedlings (GHAFFOOR; SHAH; WASEEM, thiamine (1.0 mg L-1), gibberellic acid (0.25 mg L-1),
2003). These results corroborate those observed by and sucrose (20 g L-1) under constant shaking.
Badoni and Chauhan (2009), who reported greater Similar result was observed for the organogenesis of
development and multiplication of potato seedlings potato meristematic apices (PEREIRA; FORTES,
when the culture medium was supplemented with 2004b), in which the liquid culture medium had a
0.01 mg L-1 of naphthalene-acetic acid and 0.25 mg higher multiplication rate when compared with the
L-1 of gibberellic acid. semi-solid medium.
Among the classes of plant regulators that However, a disadvantage of the cultivation
are not widely used in in vitro cultivation, jasmonic in liquid medium is the occurrence of
acid, salicylic acid, ethylene, polyamines and hyperhydricity. Hyperhidric structures were
brassinosteroids deserve attention. It is postulated reported by Rosell et al. (1987) and Park et al.
that supplementation of the culture medium and/or (2004) when studying potato micropropagation.
the pretreatment of potato mother plants with Nevertheless, the authors observed that the
jasmonic acid promotes in vitro tuberization and formation of these structures can be avoided by
improves the quality of microtubers (PRUSKI; transferring the culture to solid medium. Media are
ASTATKIE; NOWAK, 2002; SARKAR; generally polymerized with agar or phytagel, though
PANDEY; SHARMA, 2006). Moreover, this many alternative gelling agents, as Lallemantia
growth regulator confers higher rates of potato royleana seeds, are proposed to reduce the costs of
micropropagation, although not always a potato in vitro micropropagation (JABEEN;

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KAUSAR; SHAH, 2017). Besides the consistency, for most of the materials, the formation of
the concentration of salts, the type and the dose of microtubers under short photoperiod conditions (10
cytokinins present in the medium also affect the h photoperiod with irradiance of 12 µmol m–2 s–1)
formation of hyperhidric structures (PALMA et al., and low temperature (20 ± 2°C during the day and
2011). 18 ± 2°C at night). In relation to the source and dose
Important aspects concerning incubation of of carbon in the culture medium, 6% of sucrose and
micropropagated potato plants are addressed in 4% of maltose were considered suitable for the
some research. Those range from light sources potato in vitro tuberization, for cultivars Agria and
(ROCHA; OLIVEIRA; SCIVITTARO, 2015), Justine, respectively (ALTINDAL; KARADOGAN,
growth room ventilation (VENKATASALAM et al., 2010). Thus, genotype x environment interactions
2015), and culture vessels (SHUKLA et al., 2017) indicate the importance of developing specific
until changes in plantlets profiles (GUPTA; protocols for each potato cultivar in order to
PATTANAYAK, 2017; LOYOLA-VARGAS; maximize micropropagation, resulting in the
OCHOA-ALEJO, 2018). availability of sufficient and high quality material
for farmers.
Tuberization
The transition between in vitro Production of indexed seed potato
multiplication and the establishment of this In Brazil, the official production process of
solanaceous in the field is known as pathogen-free seed potatoes begins with the in vitro
microtuberization (NISTOR et al., 2010). In this production of plants from the shoot tip culture. The
case, microtubers have been used as an alternative methodology for shoot-tip propagation depends, as
method for national and international distribution of in any micropropagation process as previously
potato germplasm and as pathogen-free starting described, on asepsis, on the genotype of the plant
material for seed multiplication programs (MUÑOZ, material, on the explant itself, and on the
1996). These small tubers have some advantages in composition of the culture medium in which the
relation to in vitro seedlings, such as: easy material will be inoculated.
distribution and handling of the material due to its Shoot tips are isolated and inoculated in
reduced size, greater resistance to transport medium containing more cytokinin than auxin for
conditions, and possibility of dark storage under low differentiation into aerial parts. Subsequently, these
temperatures for longer periods than that of multiplicated shoots are transferred to the medium
micropropagated seedlings (TOVAR et al., 1985; containing only cytokinin (or not). A shoot tip can
McCOWN; JOYCE, 1991). In addition, microtubers generate from 500 to more than 1000 plants,
can be used as an explant source in studies on depending on the cultivar or even on the shoot tip.
genetic transformation (SNYDER; BELKNAP, Bandinelli et al. (2013) reported that the 50%
1993). reduction in the MS salts concentration during
Several protocols to induce in vitro multiplication increases the ex vitro survival rate in
tuberization have already been developed, such as Asterix, Macaca, and SMINIA793101-3 potato
that of Wang and Hu (1982), which is adopted by clones. At the establishment and multiplication
the International Potato Center (Centro stages, plants are kept in a growth chamber, with
Internacional de la Papa - CIP) (TOVAR et al., light (14-16 h photoperiod) and temperature (25-
1985; DODDS; SILVA-RODRIGUEZ; TOVAR, 27ºC) control (PEREIRA; FORTES, 2004b).
1992). This protocol consists of adding 6- Seedlings can be transferred to a medium richer in
benzylaminopurine, chlorocholine chloride and sucrose for in vitro tuberization. However, this
sucrose to the liquid culture medium. Tuberization procedure is not widely adopted. Instead,
can also be achieved in bioreactor systems, even tuberization in a tray or plate with sterile substrate
resulting in greater yields and less hyperhydricity under anti-aphid conditions has been more
rates compared to the liquid culture (RAHMAN et frequently used. Bandinelli et al. (2013) recommend
al., 2015). The in vitro microtuber, subjected to the mineral substrate (coarse sand), as it favors the
proper stimuli, can further be directly seeded in acclimatization of the plants, increasing the survival
open field (SHIN et al., 2018). rate in the field. However, according to Daniels,
Microtubers ideal growing conditions Pereira and Fortes (2000), the production of seed
(photoperiod, temperature, and nutrient source and tubers in solid matrix presents low productive
dose) may vary according to the genotype under efficiency, since three to five tubers per plant are
study. In this context, Gopal, Minocha and Dhaliwal produced.
(1998) evaluated 22 potato genotypes and verified,

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Soil-free (or non-solid) technologies have obtained from S1 seed potatoes, or from higher
gained attention and replaced the conventional categories (MAPA, 2012).
production system of seed tubers. Factor et al. In addition to the tuber, the shoot can also
(2007) reported that these technologies prevent the be used as propagative material in the production of
contact of seed tubers with soil phytopathogens, basic and certified seed potatoes (MOREIRA,
facilitates cleaning after the production cycle, 2008), reducing costs with the production or
enables staggered harvesting, standardizes the acquisition of seed potatoes (SILVA et al., 2011;
tubers, and guarantees high phytosanitary quality VIRMOND; KAWAKAMI; DIAS, 2011). The
and higher multiplication rate. Soil-free Normative Instruction No. 32 of November 20,
technologies include hydroponics, nutrient film 2012 establishes that certified seedlings in the form
technique, deep flow technique, and aeroponics. of sprouts can only be produced using basic seed
Among them, aeroponic systems have emerged as a potatoes (MAPA, 2012). In production of seedlings,
possibility in many developing countries, such as minitubers and basic seed potatoes, their indexation
Brazil (FACTOR et al., 2007), Peru (OTAZÚ, by the ELISA test is mandatory, in order to confirm
2010), China (WANG et al., 2017), and Uganda the absence of viruses.
(KAKUHENZIRE et al., 2017). Briefly, it consists
of spraying a nutrient solution to the suspended Somatic embryogenesis
plant roots and draining it back to the reservoir Several studies have reported the formation
tanks until the next cycle of misting. of potato somatic embryos in solid culture medium
After cultivation, seed potatoes are stored at from tuber discs (BRAGDØ-AAS, 1977; NASSAR
ambient conditions for 10 to 14 days for peel et al., 2011), nodal segments (GARCÍA;
suberification and further stored for six to eight MARTÍNEZ, 1995; SEABROOK; DOUGLASS;
months, at 3 to 4ºC, for dormancy overcome and for TAI, 2001; VARGAS; GARCIA; OROPEZA, 2005;
the sprouting of the largest number of eyes. After VARGAS et al., 2008; SHARMA et al., 2007,
this period, tubers are able to produce seed potatoes 2008b), and leaf tissues (JAYASREE et al., 2001;
in the field, in isolated areas with low incidence of KAUR; REDDY; KUMAR, 2018). The formation
insect vectors, meeting certification standards of these structures in liquid medium was first
(DUTRA et al., 2010). described by Fiegert, Mix-Wagner and Vorlop
The production and commercialization of (2000).
seed potatoes within the Brazilian territory follow Growth regulators content and ratio are key
rules that aim at ensuring the genetic identity and factors on callus induction and somatic embryos
health of this plant material, which are determined initiation and development. The molecular and
by the Ministry of Livestock and Food Supply biochemical mechanisms behind these phenomena
(MAPA). According to the Normative Instruction however are yet to be elucidated. For some
No. 32 of November 20, 2012, of MAPA, seed information, consult Nassar, Kubow and Donnelly
potatoes should be produced in the following (2015). At the first stage, auxins are required to
categories: genetic seed potatoes; basic seed potato induce callusing but must be removed during
(G0, G1, G2 and G3); certified seed potato of first embryo initiation due to their suppressive effect.
generation (C1); certified seed potato of second The most frequent used is the 2,4-
generation (C2); S1 and S2. The basic seed potato dichlorophenoxyacetic acid (2,4-D) (BORDALLO
G0 should be obtained from the genetic seed potato et al., 2004; VARGAS; GARCIA; OROPEZA,
or from the basic potato seedling, cultivated in 2005; VARGAS et al., 2008; SHARMA; MILLAM,
protected environment. The basic seed potato G1 2004; SHARMA et al., 2007, 2008a, b; NASSAR et
should be obtained from the basic seed potato G0; al., 2011; SABETI; ZARGHAMI; EBRAHIM,
the basic seed potato G2 should be obtained from 2013). Embryo formation is later induced by a range
the basic seed potato G1; and basic seed potato G3 of growth regulators, including zeatin and
should be obtained from the basic seed potato G2. gibberellin (GA3) (FIEGERT; MIX-WAGNER;
Certified seed potatoes of first generation (C1) VORLOP, 2000; JAYASREE et al., 2001;
should be obtained from basic or certified seed SEABROOK; DOUGLASS, 2001; SEABROOK;
potatoes, whereas certified seed potatoes C2 can be DOUGLASS; TAI, 2001; VARGAS; GARCIA;
obtained from certified seed potatoes C1, basic seed OROPEZA, 2005; VARGAS et al., 2008;
potatoes, or from certified seedlings. S1 seed SHARMA; MILLAM, 2004; SHARMA et al.,
potatoes should be obtained from certified seed 2007; SHARMA; SARKAR; PANDEY, 2010;
potatoes C2 or above; and S2 seed potato should be NASSAR et al., 2011).

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Jayasree et al. (2001) established an dihaploid potato genotypes. The protoplasts were
efficient protocol to induce somatic embryos and isolated from S. tuberosum mesophilic cells and
regenerate potato seedlings, cv. Jyothi, from leaf from the wild diploid species S. brevidens, and
segments. Initially, leaves are grown in MS medium fused together at high pH and high calcium
supplemented with 2,4-D and benzyladenine (BA). concentrations. Putative hybrids were then selected
The embryogenic calli, obtained from the borders of by the phenotype from regenerated shoots, and of
the leaf segments, are transferred to MS medium these, 11 somatic hybrids were identified in function
containing zeatin and BA, resulting in high of their isoenzymatic patterns and morphological
formation of somatic embryos. Embryos mature in traits.
the same induction medium, and are transferred to By means of somatic hybridization, several
MS medium without growth regulators for seedling resistance genes from wild species have already
regeneration when they reach the cotyledonary been introduced into potato, including those that
stage. confer tolerance to herbicide (BINDING et al.,
Seabrook and Douglass (2001) also reported 1982), resistance to leafroll virus (AUSTIN; BAER;
the in vitro induction of somatic embryos of 18 HELGESON, 1985), tolerance to frost (ROKKA et
tetraploid potato cultivars from several explants al., 1998), tolerance to late blight disease (SHI et al.,
(nodal segments, leaves, microtubers, and roots). 2006), and resistance to insects (CHENG;
These authors observed differences in induction SAUNDERS; SINDEN, 1995; BUTLER et al.,
capacity depending on the genotype of the evaluated 2011; DIAZ-MONTANO et al., 2014). Specific
materials, varying from 2.2 to 44.5 somatic reviews of somatic hybridization in potato can be
embryos. found in Pandey et al. (2010), Cho and Park (2014)
This technique, when applied to potato crop, and Rokka (2015).
is a potentially effective method in the production of
synthetic seeds. Synseeds produce virus-free, true- In vitro preservation and cryopreservation
to-type materials that are easy to handle and The maintenance of in vitro collections has
transport (POND; CAMERON, 2003; BORDALLO been considered an alternative method to the
et al., 2004; SHARMA et al., 2007). However, there conservation of germplasm, especially for
has been relatively little progress on this application vegetative propagated species. In the case of potato,
due to problems of somaclonal variation and the minimum in vitro growth occurs by the control
requirement of bioreactors for large-scale of incubation conditions, such as low temperatures,
production of somatic embryos followed by their low light intensity, and mixed photoperiod
encapsulation and field planting (FIEGERT; MIX- (WESTCOTT, 1981a; GOPAL; CHAUHAN, 2010),
WAGNER; VORLOP, 2000; NYENDE et al., 2002; or by the addition of substances to the culture
SCHAFER-MENUHR; MIX-WAGNER; medium, especially growth retardants and osmotic
VORLOP, 2003, BAMBERG et al., 2016). Thus, stress inducers (WESTCOTT, 1981b; FORTES;
the development of reliable systems of somatic PEREIRA, 2001; GOPAL; CHAUHAN, 2010).
embryogenesis for this solanaceous plant is still Sarkar and Naik (1998) studied the effect of
incipient (SHARMA; MILLAM, 2004), and few simultaneous manipulation of incubation conditions
studies are found in the literature regarding the and culture medium on the in vitro preservation of
occurrence and performance of somaclones in four potato genotypes. The combination of 40 g L-1
greenhouse (SEABROOK; DOUGLASS, 2001) and sucrose, 20 g L-1 mannitol, and 16 h photoperiod
in field (NASSAR, 2009; NASSAR et al., 2014) and provided better conservation of potato microplants
the genetic mechanisms that regulate this process for up to 30 months without the need for
(SHARMA et al., 2008a, b). subcultivation. Alternatively to mannitol, ancimidol
- a growth retardant - can be used (SARKAR;
Protoplast culture CHAKRABARTI; NAIK, 2001). At CIP, more than
Protoplast technology has focused 4,000 potato accessions are kept in culture medium
especially on the generation of interspecific hybrids with 40 g L-1 sorbitol and under low temperature (6-
between wild and cultivated species that cannot be 8ºC) (NIINO; ARIZAGA, 2015), as similarly
produced via conventional hybridization (DAVEY observed at the US Potato Genebank (30 g L-1
et al., 2005). In potato culture, wild Solanum species sorbitol, 8-10ºC) (BAMBERG et al., 2016). In the
are commonly used to introduce desirable traits to case of liquid culture medium, González-Benito,
commercial genotypes. Ramírez and Aranda (2004) reported that
In 1987, Fish, Karp and Jones developed a supplementation of MS medium with 2% sucrose
protoplast culture technique applicable to several

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and incubation at 10ºC allows the maintenance of genotypes the cryopreserved shoot tips presented a
potato seedlings for about 2-3 years. significant decrease in regeneration.
The choice of the conservation method to be
used should consider the occurrence of somaclonal CONCLUSIONS
variation, which may also be associated with the
nature of the plant material and to the substances Plant tissue culture techniques have been
employed (WITHERS; ENGELMANN, 1998). This widely used as an alternative to solve the lack of
genetic variation needs to be evaluated at the end of propagating material of potatoes with high
the whole process in order to detect possible threats phytosanitary quality. Among these techniques,
to the genetic stability of plant material. As an micropropagation stands out, whose clonal
example, Harding (1994) detected hypermethylation multiplication protocols have already been well
of potato microplants’ DNA maintained for long established, and whose process is considered the
periods under osmotic stress promoted by the use of main route of production of pathogen-free seed
mannitol. Conversely, Sarkar, Chakrabarti and Naik potatoes.
(2001) detected no genetic instability after 16 The use of somatic embryos has long been
months of conservation by means of Random considered promising alternative to conventional
Amplified Polymorphic DNA (RAPD) markers, potato propagules; however, little has been achieved
when evaluating the effects of ancimidol on the in the development of synthetic seeds for this
conservation of potato microplants. solanaceous plant. Thus, aspects concerning the
Long-period in vitro preservation may not application of somatic embryogenesis in this species
guarantee the genetic stability of the still require further studies. In addition, more studies
micropropagated material. Thus, cryopreservation is should address the viability of potato genetic
a more reliable alternative. In addition to long-term transformation via culture and protoplast fusion.
storage of potato germplasm, cryotherapy can In addition to large-scale clonal production
simultaneously be applied as an alternative method of indexed plants, in vitro techniques can be
for virus elimination and production of indexed employed for medium-term preservation and safe
plants (WANG et al., 2006; KACZMARCZYK; exchange of potato germplasm. Thus,
ROKKA; KELLER, 2011). cryopreservation can be considered an additional
The first report on the use of this technique tool for genetic material conservation.
in potatoes was in the late 1970s (BAJAJ, 1977), Finally, the use of the products generated by
and from that work, different methods have been scientific research by the different segments in the
developed using slow or ultrafast freezing potato production chain is directly dependent on
(TOWILL, 1984; BENSON; HARDING; SMITH, regular investments in the sector, and on the
1989). In 1995, CIP started to test a constant technical training of the professionals
cryopreservation method by vitrification. The involved in research. Only then will potato be
process consisted of dehydrating potato meristems assisted with technology capable of guaranteeing
in concentrated sugar solutions and cryoprotectors highly productive crops.
and rapidly freezing them using liquid nitrogen
(GOLMIRZAIE; PANTA, 2000). Other techniques ACKNOWLEDGEMENTS
of vitrification have already been tested for several
potato cultivars, such as the encapsulation- This research was supported by the
vitrification method (HIRAI; SAKAI, 1999; HIRAI, Coordenação de Aperfeiçoamento de Pessoal de
2011), the droplet-vitrification (KIM et al., 2006; Nível Superior – CAPES, Conselho Nacional de
YOON et al., 2006), and the V-cryo-plate Desenvolvimento Científico e Tecnológico – CNPq,
(YAMAMOTO et al., 2013). and Fundação de Amparo à Pesquisa do Estado de
The viability of potato shoot tips maintained Minas Gerais – FAPEMIG. The first author would
for long periods in liquid nitrogen has also been like to thank CAPES and Programa de Pós-
reported (MIX-WAGNER; SHUMACHER; Graduação em Agronomia (PPGA-UFU) for the
CROSS, 2003). These authors compared the financial support.
regeneration of 51 cultivars in long-term (3 to 8
years) and short-term preservation. Only for three

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Application of tissue… MORAIS, T. P. et al.

RESUMO: A batata é a cultura não-cereal mais importante do mundo e, portanto, uma das principais fontes de
alimento para a humanidade. Sua multiplicação convencional é assexuada utilizando o próprio tubérculo, o que permite o
acúmulo e a difusão de patógenos para novas áreas de cultivo, comprometendo a produtividade desta solanácea e
ameaçando a manutenção de genótipos de interesse comercial ou para fins de melhoramento. Devido à inviabilidade de
utilização das sementes botânicas, a conservação e o intercâmbio de germoplasma dessa espécie por meio de métodos
convencionais torna-se inviável. Em todas as regiões produtoras de batata, a demanda por tubérculos de alta qualidade tem
sido primordial para garantir a produção das lavouras. Dessa forma, técnicas biotecnológicas baseadas na cultura de
tecidos são de suma importância. A cultura de tecidos vegetais oferece métodos alternativos de propagação através das
técnicas in vitro que proporcionam a produção e multiplicação de material com alta sanidade. Dessa maneira, esta revisão
visa sumarizar o histórico e panorama atual das aplicações da cultura de tecidos em batata. Além da multiplicação clonal,
essa ferramenta biotecnológica fornece material inicial indexado para programas de melhoramento e de produção
certificada de batata-semente e facilita o intercâmbio e a conservação de germoplasma. Por tudo isso, o emprego destas
técnicas na cadeia produtiva da batata proporciona benefícios diretos aos produtores, uma vez que fornece material
propagativo com elevada qualidade genética e fitossanitária.

PALAVRAS-CHAVE: Solanum tuberosum L. Cultivo in vitro. Batata-semente. Erradicação de

patógenos.

REFERENCES

AGRAMONTE, D.; JIMÉNEZ, F.; PÉREZ, M.; GUTIÉRREZ, O.; RAMIREZ, D.; NUÑEZ, M. Empleo de
sustancias bioestimuladoras (Biobras-6 y Biobras-16) en la fase de adaptación de vitroplantas de papa
(Solanum tuberosum L.). La Habana: INCA, 1996. 162p.

ALI, S.; KHAN, N.; NOUROZ, F.; ERUM, S.; NASIM, W.; SHAHID, M. A. In vitro effects of GA3 on
morphogenesis of CIP potato explants and acclimatization of plantlets in field. In Vitro Cellular and
Developmental Biology Plant, v. 54, n. 1, p. 104-111, 2018. https://doi.org/10.1007/s11627-017-9874-x

ALTINDAL, D.; KARADOGAN, T. The effect of carbon sources on in vitro microtuberization of potato
(Solanum tuberosum L.). Turkish Journal of Field Crops, v. 15, n. 1, p. 7-11, 2010.

AUSTIN, S.; BAER, M. A.; HELGESON, J. P. Transfer of resistance to Potato leaf roll virus from Solanum
brevidens into Solanum tuberosum by somatic fusion. Plant Science, v. 39, p. 75-82, 1985.
https://doi.org/10.1016/0168-9452(85)90195-5

ÁVILA, A.; PEREYRA, S. M.; COLLINO, D. J.; ARGÜELLO, J. A. Effects of nitrogen source on growth and
morphogenesis of three micropropagated potato cultivars. Potato Research, v. 37, n. 2, p. 161-168, 1994.
https://doi.org/10.1007/BF02358717

BADONI, A.; CHAUHAN, J. S. Effect of growth regulators on meristem-tip development and in vitro
multiplication of potato cultivar ‘kufri himalini’. Nature and Science, v. 7, n. 9, p. 31-34, 2009.

BADR, A.; ANGERS, P.; DESJARDINS, Y. Metabolic proofing of photoautotrophic and photomixotrophic
potato plantles (Solanum tuberosum) provides new insights into acclimatization. Plant Cell, Tissue and Organ
Culture, v. 107, p. 13-24, 2011. https://doi.org/10.1007/s11240-011-9951-5

BAJAJ, Y. P. S. Initiation of shoots and callus from potato-tuber sprouts and axillary buds frozen at -196ºC.
Crop Improvement, v. 4, p. 48-53, 1977.

BAMBERG, J. B.; MARTIN, M. W.; ABAD, J.; JENDEREK, M. M., TANNER, J.; DONNELLY, D. J.;
NASSAR, A. M. K.; VEILLEUX, R. E.; NOVY, R. G. In vitro technology at the US Potato Genebank. In
Vitro Cellular and Developmental Biology Plant, v. 52, n. 3, p. 213-225, 2016.
https://doi.org/10.1007/s11627-016-9753-x

Biosci. J., Uberlândia, v. 34, n. 4, p. 952-969, July/Aug. 2018

 960
Application of tissue… MORAIS, T. P. et al.

BANDINELLI, M. G.; BISOGNIN, D. A.; GNOCATO, F. S.; MAMBRIN, R. B.; SAUSEN, D.; NICOLOSO,
F. T. Concentração dos sais e da sacarose do meio MS na multiplicação in vitro e na aclimatização de batata.
Horticultura Brasileira, v. 31, n. 2, p. 242-247, 2013. https://doi.org/10.1590/S0102-05362013000200011

BASERA, M.; CHANDRA, A.; KUMAR, V. A.; KUMAR, A. Effect of brassinosteroids on in vitro
proliferation and vegetative growth of potato. The Pharma Innovation Journal, v. 7, n. 4, p. 4-9, 2018.

BENSON, E. E.; HARDING, K.; SMITH, H. Variation in recovery of cryopreserved shoot tips of Solanum
tuberosum exposed to different pre- and post-freeze light regimes. CryoLetters, v. 10, p. 323-344, 1989.

BINDING, H.; JAIN, S. M.; FINGERINGER, J.; MORDHORST, G.; NEHLS, R.; GRESSEL, J. Somatic
hybridization of an atrazine resistant biotype of Solanum nigrum with Solanum tuberosum. Theoretical and
Applied Genetics, v. 63, n. 3, p. 273-277, 1982. https://doi.org/10.1007/BF00304007

BORDALLO, P. N.; SILVA, D. H.; MARIA, J.; CRUZ, C. D.; FONTES, E. P. Somaclonal variation on in
vitro callus culture potato cultivars. Horticultura Brasileira, v. 22, p. 300-304, 2004.
https://doi.org/10.1590/S0102-05362004000200027

BRAGDØ-AAS, M. Regeneration of plants from callus of potato tubers. Acta Horticulturae, v. 78, p. 133-
138, 1977. https://doi.org/10.17660/ActaHortic.1977.78.16

BUTLER, C. D.; GONZALEZ, B.; MANJUNATH, K. L.; LEE, R. F.; NOVY, R. G.; MILLER, J. C.;
TRUMBLE, J. T. Behavioral responses of adult potato psyllid, Bactericera cockerelli (Hemiptera: Triozidae),
to potato germplasm and transmission of Candidatus Liberibacter psyllaurous. Crop Protection, v. 30, p.
1233-1238, 2011. https://doi.org/10.1016/j.cropro.2011.05.006

CEARLEY, T. A.; BOLYARD, M. G. Regeneration of Solanum tuberosum cv. Katahdin from leaf explants in
vitro. American Journal of Potato Research, v.74, n.2, p.125-129, 1997. https://doi.org/10.1007/BF02851558

CHENG, J. J.; SAUNDERS, A.; SINDEN, S. L. Colorado potato beetle resistant somatic hybrid potato plants
produced via protoplast electrofusion. In Vitro Cellular and Developmental Biology Plant, v. 31, n. 2, p. 90-
95, 1995. https://doi.org/10.1007/BF02632243

CHO, K. S.; PARK, T. H. Potato breeding via protoplast fusion. Journal of Plant Biotechnology, v. 41, p. 65-
72, 2014 (in Korean with English abstract). https://doi.org/10.5010/JPB.2014.41.2.65

DANIELS, J.; PEREIRA, A. S.; FORTES, G. R. L. Verticalização da produção de batata-semente por

produtores de agricultura familiar no Rio Grande do Sul. Pelotas: Embrapa Clima Temperado, 2000. 4p.

DAVEY, M. R.; ANTHONY, P.; POWER, J. B.; LOWE, K. C. Plant protoplasts: status and biotechnological
perspectives. Biotechnology Advances, v. 23, n. 2, p. 131-171, 2005.
https://doi.org/10.1016/j.biotechadv.2004.09.008

DIAZ-MONTANO, J.; VINDIOLA, B. G.; DREW, N.; NOVY, R. G.; MILLER, J. C.; TRUMBLE, J. T.
Resistance of selected potato genotypes to the potato psyllid (Hemiptera: Triozidae). American Journal of
Potato Research, v. 91, p. 363-367, 2014. https://doi.org/10.1007/s12230-013-9356-6

DODDS, J. H.; SILVA-RODRIGUEZ, D.; TOVAR, P. Micropropagation of potato (Solanum tuberosum L.).
In: BAJAJ, Y. P. S. (Ed.). Biotechnology in Agriculture and Forestry. High-Tech and Micropropagation III.
New York, Heidelberg, Berlin: Springer, 1992. v. 19, p. 91-205. https://doi.org/10.1007/978-3-662-07770-2_6

DONNELLY, D. J.; COLEMAN, W. K.; COLEMAN, S. E. Potato microtuber production and performance: a
review. American Journal of Potato Research, v. 80, n. 2, p. 103-115, 2003.
https://doi.org/10.1007/BF02870209

Biosci. J., Uberlândia, v. 34, n. 4, p. 952-969, July/Aug. 2018

 961
Application of tissue… MORAIS, T. P. et al.

DUTRA, L. F.; MAYER, K. C. A.; SILVA, N. D. G.; NINO, A. F. P.; SILVA, F. O. X.; VIEIRA, F. C. B.
Protocolo de Micropropagação de Plantas. I - Batata. Pelotas: Embrapa Clima Temperado, 2010. 22p.

EL-FIKI, A. I. I.; EL DIN, T. M. N.; MAHDY, A. M. M.; ALI, A. S. Elimination of Potato virus X and
comparable of microtuber productivity of some infected potatoes in vitro. Annals of Agricultural Science, v.
30, n. 1, p. 195-208, 1992.

ESNA-ASHARI, M.; VILLIERS, T. A. Plant regeneration from tuber disc of potato (Solanum tuberosum L.)
using 6-benzylaminopurine (6-BAP). Potato Research, v. 41, n. 4, p. 371-382, 1998.
https://doi.org/10.1007/BF02358969

FACTOR, T. L.; ARAÚJO, J. A. C.; KAWAKAMI, F. P. C.; IUNCK, V. Produção de minitubérculos básicos
de batata em três sistemas hidropônicos. Horticultura Brasileira, v. 25, n. 1, p. 82-87, 2007.
https://doi.org/10.1590/S0102-05362007000100016

FERAY, A.; HOURMANT, A.; BRUN, A.; PENOT, M. Effect of polyamines on morphogenesis of in vitro
potato plants (Solanum tuberosum cv. Bintje). Comptes Rendus de L'Académie des Sciences, v. 316, n. 12, p.
1446-1451, 1993.

FIEGERT, A. K.; MIX-WAGNER, G.; VORLOP, K. D. Regeneration of Solanum tuberosum L. cv. Tomensa:
induction of somatic embryogenesis in liquid culture for the production of “artificial seed”. Landbauforsch
Völkenrode, v. 50, n. 3-4, p. 199-202, 2000.

FISH, N.; KARP, A.; JONES, M. G. K. Improved isolation of dihaploid Solanum tuberosum protoplasts and
the production of somatic hybrids between dihaploid S. tuberosum and S. brevidens. In Vitro Cellular and
Developmental Biology Plant, v. 23, n. 8, p. 575-580, 1987. https://doi.org/10.1007/BF02620975

FORTES, G. R. L.; PEREIRA, J. E. S. Batata-semente pré-básica: Cultura de Tecidos. In: PEREIRA, A. S.;
DANIELS, J. (Eds). O cultivo da batata na região sul do Brasil. Brasília: Embrapa Informação Tecnológica,
2003a. p. 421-433.

FORTES, G. R. L.; PEREIRA, J. E. S. Classificação e descrição botânica. In: PEREIRA, A. S.; DANIELS, J.
(Eds). O cultivo da batata na região sul do Brasil. Brasília: Embrapa Informação Tecnológica, 2003b. p. 53-
143.

FORTES, G. R. L.; PEREIRA, J. E. S. Preservação in vitro da batata com ácido acetilsalicílico e duas fontes de
carboidrato. Pesquisa Agropecuária Brasileira, v. 36, n. 10, p. 1261-1264, 2001.
https://doi.org/10.1590/S0100-204X2001001000007

GARCÍA, E. D.; MARTÍNEZ, S. Somatic embryogenesis in Solanum tuberosum L. cv. Desiree from stem
nodal sections. Journal of Plant Physiology, v. 145, p. 526-530, 1995. https://doi.org/10.1016/S0176-
1617(11)81782-7

GHAFFOOR, A.; SHAH, G. B.; WASEEM, K. In vitro response of potato (Solanum tuberosum L.) to various
growth regulators. Biotechnology, v. 2, n. 3, p. 191-197, 2003. https://doi.org/10.3923/biotech.2003.191.197

GOLMIRZAIE, A. M.; PANTA, A. Advances in potato cryopreservation at CIP. In: ENGELMANN, F.;
TAKAGI, H. (Eds). Cryopreservation of tropical plant germplasm: current research progress and
application. Tsukuba: JIRCAS/ Rome: IPGRI, 2000. p. 250-254.

GONÇALVES, J. F.; MALDANER, J.; ROSSATO, L. V.; TABALDI, L. A.; SKREBSKY, E. C.; FARIAS, J.
G.; BISOGNIN, D. A.; NICOLOSO, F. T. Crescimento in vitro de plântulas de batata em diferentes doses de
cádmio. Ciência Rural, v. 39, n. 9, p. 2625-2628, 2009. https://doi.org/10.1590/S0103-84782009000900038

Biosci. J., Uberlândia, v. 34, n. 4, p. 952-969, July/Aug. 2018

 962
Application of tissue… MORAIS, T. P. et al.

GONZÁLEZ-BENITO, M. E. G.; RAMÍREZ, I. C.; ARANDA, J. M. L. Review. The use of cryopreservation

for germplasm conservation of vegetatively propagated crops. Spanish Journal of Agricultural Research, v.
2, n. 3, p. 341-351, 2004. https://doi.org/10.5424/sjar/2004023-88

GOPAL, J.; CHAUHAN, N. S. Slow growth in vitro conservation of potato germplasma at low temperature.
Potato Research, v. 53, n. 3, p. 141-149, 2010. https://doi.org/10.1007/s11540-010-9158-x

GOPAL, J.; MINOCHA, J. L.; DHALIWAL, H. S. Microtuberization in potato (Solanum tuberosum L.). Plant
Cell Reports, v. 17, n. 10, p. 794-798, 1998. https://doi.org/10.1007/s002990050485

GRATTAPAGLIA, D.; MACHADO, M. A. Micropropagação. In: TORRES, A. C.; CALDAS, L. S.; BUSO, J.
A. (Eds). Cultura de tecidos e transformação genética de plantas. Brasília: Embrapa-SPI, 1998. v. 1, p. 183-
260.

GUPTA, S. D.; PATTANAYAK, A. K. Intelligent image analysis (IIA) using artificial neural network (ANN)
for non-invasive estimation of chlorophyll content in micropropagated plants of potato. In Vitro Cellular and
Developmental Biology Plant, v. 53, n. 6, p. 520-526, 2017. https://doi.org/10.1007/s11627-017-9825-6

HARDING, K. The methylation status of DNA derived from potato plants recovered from slow growth. Plant
Cell, Tissue and Organ Culture, v. 37, p. 31-38, 1994. https://doi.org/10.1007/BF00048114

HIRAI, D. Gelled droplet vitrification improves recovery of cryopreserved potato germplasm. CryoLetters, v.
32, n. 4, p. 287-296, 2011.

HIRAI, D.; SAKAI, A. Cryopreservation of in vitro-grown meristems of potato (Solanum tuberosum L.) by
encapsulation-vitrification. Potato Research, v. 42, n. 1, p. 153-160, 1999.
https://doi.org/10.1007/BF02358405

IBGE - INSTITUTO BRASILEIRO DE GEOGRAFIA E ESTATÍSTICA. Levantamento sistemático da

produção agrícola: pesquisa mensal de previsão e acompanhamento das safras agrícolas no ano civil. Rio de
Janeiro: IBGE, 2017. 112p.

JABEEN, T.; KAUSAR, R.; SHAH, A. H. Development of low cost protocol for micropropagation of Solanum
tuberosum L. Bangladesh Journal of Botany, v. 46, n. 4, p. 1341-1345, 2017.

JAYASREE, T.; PAVAN, U.; RAMESH, M.; RAO, A. V.; REDDY, J. M.; SADANANDAM, A. Somatic
embryogenesis from leaf cultures of potato. Plant Cell, Tissue and Organ Culture, v. 64, n. 1, p. 13-17, 2001.
https://doi.org/10.1023/A:1010697608689

KACZMARCZYK, A.; ROKKA, V. M.; KELLER, E. R. J. Potato shoot tip cryopreservation. Potato
Research, v. 54, p. 45-79, 2011. https://doi.org/10.1007/s11540-010-9169-7

KAKUHENZIRE, R.; TIBANYENDERA, D.; KASHAIJA, I. N.; LEMAGA, B.; KIMOONE, G.; KESIIME,
V. E.; OTAZU, V.; ORTIZ, O.;BARKER, I. Improving minituber production from tissue-cultured potato
plantlets with aeroponic technology in Uganda. International Journal of Agriculture and Environmental
Research, v. 3, n. 5, p. 3948-3964, 2017.

KAUR, A.; REDDY, M. S.; KUMAR, A. Direct somatic embryogenesis of potato [Solanum tuberosum (L.)]
cultivar ‘Kufri Chipsona 2’. Plant Cell, Tissue and Organ Culture, p. 1-10, 2018.
https://doi.org/10.1007/s11240-018-1435-4

KAUR, A.; REDDY, M. S.; KUMAR, A. Efficient, one step and cultivar independent shoot organogenesis of
potato. Physiology and Molecular Biology of Plants, v. 23, n. 2, p. 461-469, 2017.
https://doi.org/10.1007/s12298-017-0418-y

Biosci. J., Uberlândia, v. 34, n. 4, p. 952-969, July/Aug. 2018

 963
Application of tissue… MORAIS, T. P. et al.

KIM, H. H.; YOON, J.; PARK, Y.; CHO, E.; SOHN, J.; KIM, T.; ENGELMANN, F. Cryopreservation of
potato cultivated varieties and wild species: critical factors in droplet vitrification. CryoLetters, v. 27, n. 4, p.
223-234, 2006.

KUMLAY, A. M. The effect of jasmonic acid on the micropropagation of potato (Solanum tuberosum L.)
under long days conditions. YYU Journal of Agricultural Sciences, v. 26, n. 1, p. 79-88, 2016.

LEIFERT, C.; WOODWARD, S. Laboratory contamination management: the requirement for microbiological
quality assurance. Plant Cell, Tissue and Organ Culture, v. 52, n. 1, p. 83-88, 1998.
https://doi.org/10.1023/A:1005905604043

LORENZEN, J. H.; NOLTE, P.; MARTIN, D.; PASCHE, J. S.; GUDMESTAD, N. C. NE-11 represents a new
strain variant class of Potato virus Y. Archives Virology, v. 153, p. 517-525, 2008.
https://doi.org/10.1007/s00705-007-0030-5

LORENZEN, J. H.; PICHIE, L. M.; GUDMESTAD, N. C.; MEACHAM, T.; SCHIEL, P. A multiplex PCR
assay to characterize Potato virus Y isolates and identify strain mixtures. Pant Disease, v. 90, p. 935-940, 2006.
https://doi.org/10.1094/PD-90-0935

LOYOLA-VARGAS, V. M.; OCHOA-ALEJO, N. An introduction to plant tissue culture: advances and

perspectives. In: LOYOLA-VARGAS, V.; OCHOA-ALEJO, N. (Eds). Plant Cell Culture Protocols:
Methods in molecular biology. New York: Humana Press, 2018. p. 3-13. https://doi.org/10.1007/978-1-4939-
8594-4 https://doi.org/10.1007/978-1-4939-8594-4_1

MADER, J. C. Polyamines in Solanum tuberosum in vitro: free and conjugated polyamines in hormone-
induced tuberization. Journal of Plant Physiology, v. 146, p. 115-120, 1995. https://doi.org/10.1016/S0176-
1617(11)81976-0

MAPA – MINISTÉRIO DA AGRICULTURA, PECUÁRIA E ABASTECIMENTO. Instrução normativa

32/2012: Normas gerais para certificação de batata-semente. Brasília, DF, 2012. 15p.

McCOWN, B.; JOYCE, P. Automated propagation of microtubers of potato. In: VASIL, I. (Ed.). Cell culture
and somatic cell genetics of plants. San Diego: Academic Press, 1991. v. 8, p. 235.
https://doi.org/10.1016/B978-0-12-715008-6.50012-9

MENEZES, C. B.; PINTO, C. A. B. P.; NURMBERG, P. L.; LAMBERT, E. S. Avaliação de genótipos de

batata (Solanum tuberosum L.) nas safras “das águas” e de inverno no sul de Minas Gerais. Ciência e
Agrotecnologia, v. 23, n. 4, p. 776-783, 1999.

MIX-WAGNER, G.; SHUMACHER, H. M.; CROSS, R. J. Recovery of potato apices after several years of
storage in liquid nitrogen. CryoLetters, v. 24, n. 1, p. 33-41, 2003.

MÖLLERS, C.; ZHANGM, S.; WENZEL, G. The influence of silver thiosulfate on potato protoplast cultures.
Plant Breeding, v. 108, n. 1, p. 12-18, 1992. https://doi.org/10.1111/j.1439-0523.1992.tb00094.x

MORE, O.; HERNANDEZ, M.; NUNEZ, M.; ESTEVEZ, A.; GONZALEZ, M. E. Empleo de analogos de
brassinosteroides en la formacion de callus embriogenicos en papa (Solanum tuberosum L.). Cultivos
Tropicales, v. 224, p. 29-35, 2001.

MOREIRA, M. A. Característica da planta e produção de batata semente básica em substrato com

diferentes materiais de propagação. 2008. 174f. Tese (Doutorado em Fitotecnia) - Universidade Federal de
Viçosa, Viçosa, Brasil.

Biosci. J., Uberlândia, v. 34, n. 4, p. 952-969, July/Aug. 2018

 964
Application of tissue… MORAIS, T. P. et al.

MUÑOZ, G. D. C. J. Propagación in vitro y microtuberización de cultivares de Solanum tuberosum L. ssp.

tuberosum Hawkes en medios liquidos. 1996. 91f. Dissertação (Mestrado em Ciências/Botânica) -
Universidad Austral de Chile, Valdivia, Chile.

MUÑOZ, M. A.; KALAZICH, J. C.; FOLCH, C. V.; ORENA, S. V.; WINKLER, A. The use and impact of
biotechnology in potato breeding: experience of the Potato Breeding Program at INIA, Chile. In: YILDIZ, M.
(Ed.). Potato: from Incas to all over the world. London: Intech Open, 2018. p. 3-17.
https://doi.org/10.5772/intechopen.72961

NAGIB, A.; HOSSAIN, S. A.; ALAM, M. F.; HOSSAIN, M. M.; ISLAM, R.; SULTANA, R. S. Virus free
potato tuber seed production through meristem culture in tropical Asia. Asian Journal of Plant Sciences, v. 2,
p. 616-622, 2003. https://doi.org/10.3923/ajps.2003.616.622

NAIK, P. S.; BUCKSETH, T. Recent advances in virus elimination and tissue culture for quality potato seed
production. In: GOSAL, S.; WANI, S. (Eds). Biotechnologies of crop improvement. Cham: Springer, 2018.
Volume 1. p. 131-158. https://doi.org/10.1007/978-3-319-78283-6_4

NASCIMENTO, L. C.; PIO-RIBEIRO, G.; WILLADINO, L.; ANDRADE, G. P. Stock indexing and Potato
virus Y elimination from potato plants cultivated in vitro. Scientia Agricola, v. 60, n. 3, p. 525-530, 2003.
https://doi.org/10.1590/S0103-90162003000300017

NASSAR, A. M. K.; ABDULNOUR, J.; LECLERC, Y.; LI, X-Q.; DONNELLY, D. J. Intraclonal selection for
improved processing of NB ‘Russet Burbank’ potato. American Journal of Potato Research, v. 88, p. 387-
397, 2011. https://doi.org/10.1007/s12230-011-9204-5

NASSAR, A. M. K.; KUBOW, S.; DONNELLY, D. J. Somatic embryogenesis for potato (Solanum tuberosum
L.) improvement. In: LI, X-Q; DONNELLY, D. J.; JENSEN, T. G. (Eds). Somatic genome manipulation:
Advances, methods, and applications. New York, Heidelberg, Dordrecht, London: Springer, 2015. p. 169-198.

NASSAR, A. M. K.; KUBOW, S.; LECLERC, Y. N.; DONNELLY, D. J. Somatic mining for phytonutrient
improvement of ‘Russet Burbank’ potato. American Journal of Potato Research, v. 91, p. 89-100, 2014.
https://doi.org/10.1007/s12230-013-9334-z

NASSAR, A. M. K. Use of somatic embryogenesis in potato (Solanum tuberosum L.) cv. Russet Burbank
improvement. 2009. 192f. Tese (Doutorado em Fitotecnia) - McGill University, Montreal, Canada.

NIINO, T.; ARIZAGA, M. V. Cryopreservation for preservation of potato genetic resources. Breeding
Science, v. 65, n. 1, p. 41-52, 2015. https://doi.org/10.1270/jsbbs.65.41

NIKITIN, M. M.; STATSYUK, N. V.; FRANTSUZOV, P. A.; DZHAVAKHIYA, V. G.; GOLIKOV, A. G.

Matrix approach to the simultaneous detection of multiple potato pathogens by real-time PCR. Journal of
Applied Microbiology, v. 124, p. 797-809, 2018. https://doi.org/10.1111/jam.13686

NISTOR, A.; CÂMPEANU, G.; ATANASIU, N.; CHIRU, N.; CARACSONIY, D. Influence of potato
genotypes on “in vitro” production of microtubers. Romanian Biotechnological Letters, v. 15, n. 3, p. 5317-
5324, 2010.

NYENDE, A. B.; SCHITTENHELM, S.; MIX-WAGNER, G.; GREEF, J. M. Synthetic potato seeds offer the
potential to improve Kenyan seed system. Landbauforsch Volk, v. 42, p. 141-148, 2002.

OTAZÚ, V. Manual on quality seed potato production using aeroponics. Lima: International Potato Center
(CIP), 2010. 44p. https://doi.org/10.4160/9789290603924

Biosci. J., Uberlândia, v. 34, n. 4, p. 952-969, July/Aug. 2018

 965
Application of tissue… MORAIS, T. P. et al.

PALMA, D.; SCHUELTER, A. R.; STEFANELLO, S.; FORTES, A. M. T. Aspectos morfofisiológicos e

controle da hiperhidricidade na cultura de tecidos vegetais. Revista Brasileira de Agrociências, v. 17, n. 2-4,
p. 174-184, 2011.

PANDEY, S. K.; SARKAR, D.; SHARMA, S.; CHANDEL, P. Integration of somatic fusion into potato
breeding: problems and perspectives. American Potato Journal, v.37, p.9-20, 2010.

PARK, S. W.; JEON, H. H.; KIM, H. S.; PARK, Y. M.; ASWATH, C.; JOUNG, H. Effect of sealed and
vented gaseous microenvironments on the hyperhydricity of potato shoots in vitro. Scientia Horticulturae, v.
99, p. 199-205, 2004. https://doi.org/10.1016/S0304-4238(03)00097-9

PAVLOVNA, U. I.; NIKOLAEVNA, K. N.; VLADIMIROVNA, D. E.; VENIAMINOVNA, S. O.;

ALEKSANDROVICH, S. D. Brassinosteroids as a factor of photosyntetic activity increase of improved
potatoes. Biosciences Biotechnology Research Asia, v. 12, n. 2, p. 1481-1485, 2015.
https://doi.org/10.13005/bbra/1807

PEREIRA, J. E. S.; FORTES, G. R. L. Multiplicación in vitro de la papa en medio sólido y líquido: influencia
de diferentes medios de cultura. Horticultura Argentina, v. 19, n. 46, p. 24, 2000.
https://doi.org/10.1590/S0100-204X2003000900003

PEREIRA, J. E. S.; FORTES, G. R. L. Protocolo para produção de material propagativo de batata em meio
líquido. Pesquisa Agropecuária Brasileira, v. 38, n. 9, p. 1035-1043, 2003a. https://doi.org/10.1590/S0100-
204X2003001100004

PEREIRA, J. E. S.; FORTES, G. R. L. Toxicidade de antibióticos no cultivo in vitro da batata em meios semi-
sólido e líquido. Pesquisa Agropecuária Brasileira, v. 38, n. 11, p. 1273-1279, 2003b.

PEREIRA, J. E. S.; FORTES, G. R. L. Produção de mudas pré-básicas de batata por estaquia a partir de plantas
micropropagadas. Horticultura Brasileira, v. 22, n. 2, p. 185-191, 2004a. https://doi.org/10.1590/S0102-
05362004000200005

PEREIRA, J. E. S.; FORTES, G. R. L. Organogênese de ápices meristemáticos de batata em meios de

isolamento e multiplicação in vitro. Horticultura Brasileira, v. 22, n. 2, p. 197-201, 2004b.
https://doi.org/10.1590/S0102-05362004000200007

PEREIRA, J. E. S.; MATTOS, M. L. T.; FORTES, G. R. Identificação e controle com antibióticos de bactérias
endofíticas contaminantes em explantes de batata micropropagados. Pesquisa Agropecuária Brasileira, v. 38,
n. 7, p. 827-834, 2003. https://doi.org/10.1590/S0100-204X2003000700006

PERL, A.; AVIV, D.; GALUN, E. Ethylene and in vitro culture of potato: suppression of ethylene generation
vastly improves protoplast yield, plating efficiency and transient expression of an alien gene. Plant Cell
Reports, v. 7, n. 6, p. 403-406, 1988.

PONDS, S.; CAMERON, H. Artificial seeds. In: THOMAS, B.; MURPHY, D. J.; MURRAY, B. G. (Eds).
Encyclopedia of applied plant sciences. Elseveir, 2003. v. 3, p. 1379-1388. https://doi.org/10.1016/B0-12-
227050-9/00217-9

PRETOVA, A.; DEDICOVA, B. Somatic embryogenesis in Solanum tuberosum L. cv. Desiree from unripe
zygotic embryos. Journal of Plant Physiology, v. 139, p. 539-542, 1992. https://doi.org/10.1016/S0176-
1617(11)80366-4

PROTACIO, C. M.; FLORES, H. E. The role of polyamines in potato tuber formation. In Vitro Cellular &
Developmental Biology - Plant, v. 28, n. 2, p. 81-86, 1992. https://doi.org/10.1007/BF02823023

Biosci. J., Uberlândia, v. 34, n. 4, p. 952-969, July/Aug. 2018

 966
Application of tissue… MORAIS, T. P. et al.

PRUSKI, K.; ASTATKIE, T.; NOWAK, J. Jasmonate effects on in vitro tuberization and tuber bulking in two
potato cultivars (Solanum tuberosum L.) under different media and photoperiod conditions. In Vitro Cellular
and Developmental Biology Plant, v. 38, p. 203-209, 2002. https://doi.org/10.1079/IVPIVP2001265

RAHMAN, Z.; ISLAM, S. M. S.; CHOWDHURY, A. N.; SUBRAMANIAM, S. Efficient microtuber

production of potato in modified nutrient spray bioreactor system. Scientia Horticulturae, v. 192, p. 369-374,
2015. https://doi.org/10.1016/j.scienta.2015.06.014

RAVNIKAR, M.; GOGALA, N. Regulation of potato meristem development by jasmonic acid in vitro.
Journal of Plant Growth Regulators, v. 9, p. 233-236, 1990. https://doi.org/10.1007/BF02041968

RAVNIKAR, M.; VILHAR, B.; GOGALA, N. Stimulatory effects of jasmonic acid on potato node and
protoplast culture. Journal of Plant Growth Regulators, v. 57, p. 151-180, 1992.
https://doi.org/10.1007/BF00193840

ROCHA, P. S. G.; OLIVEIRA, R. P.; SCIVITTARO, W. B. New light sources for in vitro potato
micropropagation. Bioscience Journal, v. 31, n. 5, p. 1312-1318, 2015. https://doi.org/10.14393/BJ-
v31n5a2015-26601

ROKKA, V. M. Protoplast technology in genome manipulation of potato through somatic cell fusion. In: LI,
X.; DONNELLY, D. J.; JENSEN, T. G. (Eds). Somatic genome manipulation. New York: Springer, 2015. p.
217-235. https://doi.org/10.1007/978-1-4939-2389-2_10

ROKKA, V. M.; TAURIAINEN, A.; PIETILÄ, L.; PEHU, E. Interspecific somatic hybrids between wild
potato Solanum acaule Bitt. and anther-derived dihaploid potato (Solanum tuberosum L.). Plant Cell Reports,
v. 18, n. 1, p. 82-88, 1998. https://doi.org/10.1007/s002990050536

ROSELL, G.; DE-BERTOLDI, F. G.; TIZIO, R.; DE-BERTOLDI, F. G. In vitro mass tuberization as a
contribution to potato micropropagation. Potato Research, v. 30, n. 1, p. 111-116, 1987.
https://doi.org/10.1007/BF02357689

ROSTAMI, F.; EHSANPOUR, A. A. The effect of silver thiosulfate (STS) on chlorophyll content and the
antioxidant enzymes activity of potato (Solanum tuberosum L.). Journal of Cell and Molecular Research, v.
2, n. 1, p. 29-34, 2010.

SABETI, M.; ZARGHAMI, R.; EBRAHIM, Z. M. Effects of explants and growth regulators on callogenesis
and somatic embryogenesis of Agria potato cultivar. International Journal of AgriScience, v. 3, p. 213-221,
2013.

SALEHI, H.; KHOSH-KHUI, M. A simple procedure for disinfection of “baby masquerade” miniature rose
explants. Scientia Horticulturae, v. 68, n. 1-4, p. 145-148, 1997. https://doi.org/10.1016/S0304-
4238(96)00978-8

SANTIAGO, G., REINIGER, L. R. S., HANAUER, J. G., LOPES, S. J., DEPRÁ, M. S. Variação somaclonal
nos descritores mínimos de tubérculo em batata. Ciência Rural v. 42, n. 2, p. 197-202, 2012.
https://doi.org/10.1590/S0103-84782012000200002

SARKAR, D.; CHAKRABARTI, S. K.; NAIK, P. S. Slow-growth conservation of potato microplants: efficacy
of ancymidol for long-term storage in vitro. Euphytica, v. 117, p. 133-142, 2001.
https://doi.org/10.1023/A:1004087422743

SARKAR, D.; KAUSHIK, S. K.; NAIK, P. S. Minimal growth conservation of potato microplants: silver
thiosulfate reduces ethylene-induced growth abnormalities during prolonged storage in vitro. Plant Cell
Reports, v. 18, n. 11, p. 897-903, 1999. https://doi.org/10.1007/s002990050681

Biosci. J., Uberlândia, v. 34, n. 4, p. 952-969, July/Aug. 2018

 967
Application of tissue… MORAIS, T. P. et al.

SARKAR, D.; NAIK, P. S. Factors affecting minimal growth conservation of potato microplants in vitro.
Euphytica, v. 102, n. 2, p. 275-280, 1998. https://doi.org/10.1023/A:1018309300121

SARKAR, D.; PANDEY, S. K.; SHARMA, S. Cytokinins antagonize the jasmonates action on the regulation
of potato (Solanum tuberosum L.) tuber formation in vitro. Plant Cell, Tissue and Organ Culture, v. 87, p.
285-295, 2006. https://doi.org/10.1007/s11240-006-9166-3

SCHAFER-MENUHR, A.; MIX-WAGNER, G.; VORLOP, K. D. Regeneration of plants from cell suspension
cultures and encapsulated cell suspension cultures of Solanum tuberosum L. Clarissa. Landbauforsch Volk, v.
53, p. 53-59, 2003.

SCHERWINSKI-PEREIRA, J. E. Contaminações microbianas na cultura de células, tecidos e órgãos de

plantas. Brasília: Embrapa Informação Tecnológica, 2010. v. 1. 8p.

SEABROOK, J. E. A.; DOUGLASS, L. K. Somatic embryogenesis on various potato tissues from a range of
genotypes and ploidy levels. Plant Cell Reports, v. 20, n. 3, p. 175-182, 2001.
https://doi.org/10.1007/s002990000305

SEABROOK, J. E. A.; DOUGLASS, L. K.; TAI, G. C. C. Segregation for somatic embryogenesis on stem
internode explants from potato seedlings. Plant Cell, Tissue and Organ Culture, v. 65, p. 69-73, 2001.
https://doi.org/10.1023/A:1010632121996

SHAHZAD, A.; SHARMA, S.; PARVEEN, S.; SAEED, T.; SHAHEEN, A.; AKHTAR, R.; YADAV, V.;
UPADHYAY, A.; AHMAD, Z. Historical perspective and basic principles of plant tissue culture. In: ABDIN,
M.; KIRAN, U.; KAMALUDDIN, A. A. (Eds). Plant Biotechnology: principles and applications. Singapore:
Springer, 2017. p.1-36.SHARMA, S. K.; BRYAN, G. J.; WINFIELD, M. O.; MILLAM, S. Stability of potato
(Solanum tuberosum L.) plants regenerated via somatic embryos, axillary bud proliferated shoots, microtubers
and true potato seeds: a comparative phenotypic, cytogenetic and molecular assessment. Planta, v. 226, p.
1449-1458, 2007. https://doi.org/10.1007/978-981-10-2961-5_1

SHARMA, S. K.; MILLAM, S. Somatic embryogenesis in Solanum tuberosum L.: a histological examination
of key developmental stages. Plant Cell Reports, v. 23, n. 3, p. 115-119, 2004. SHARMA, S. K.; MILLAM,
S.; HEIN, I.; BRYAN, G. J. Cloning and molecular characterization of a potato SERK gene transcriptionally
induced during initiation of somatic embryogenesis. Planta, v. 228, n. 2, p. 319-330, 2008a.
https://doi.org/10.1007/s00425-008-0739-8

SHARMA, S. K.; MILLAM, S.; HEDLEY, P. E.; McNICOL, J.; BRYAN, G. J. Molecular regulation of
somatic embryogenesis in potato: an auxin led perspective. Plant Molecular Biology, v.68, n.1, p.185-201,
2008b. https://doi.org/10.1007/s11103-008-9360-2

SHARMA, S.; SARKAR, D.; PANDEY, S. K. Phenotypic characterization and nuclear microsatellite analysis
reveal genomic changes and rearrangements underlying androgenesis in tetraploid potatoes (Solanum
tuberosum L.). Euphytica, v. 171, p. 313-326, 2010. https://doi.org/10.1007/s10681-009-9983-7

SHI, Y. Z.; CHEN, Q.; LI, H. Y.; BEASLEY, D.; LYNCH, D. R. Somatic hybridization between Solanum
tuberosum and S. cardiophyllum. Canadian Journal of Plant Science, v. 86, n. 2, p. 539-545, 2006.
https://doi.org/10.4141/P05-076

SHIN, K.; CHO, W.; JEON, E.; JEONG, H.; SEOL, E.; PARK, J.; LEE, Y. Pretreatment method for directly
seeding in vitro microtuber. United States Patent: US 2018/0092317 A1, 2018.

SHUKLA, M. R.; SINGH, A. S.; PIUNNO K.; SAXENA, P. K.; JONES, A. M. P. Application of 3D printing
to prototype and develop novel plant tissue culture systems. Plant Methods, v. 13, n. 6, p. 1-10, 2017.
https://doi.org/10.1186/s13007-017-0156-8

Biosci. J., Uberlândia, v. 34, n. 4, p. 952-969, July/Aug. 2018

 968
Application of tissue… MORAIS, T. P. et al.

SILVA, E. C.; PINTO, C. A.; SOUZA-DIAS, J. A. C.; ARAÚJO, T. H. Uso de reguladores de crescimento em
brotos destacados de batata-semente. Horticultura Brasileira, v. 29, n. 4, p. 504-509, 2011.
https://doi.org/10.1590/S0102-05362011000400010

SILVA FILHO, J. B.; FONTES, P. C. R.; CECON, P. R.; MCGIFFEN JR, M. E. Evaluation of “UFV
Aeroponic System” to produce basic potato seed minitubers. American Journal of Potato Research, [Online],
2018.

SNYDER, G. W.; BELKNAP, W. R. A modified method for routine Agrobacterium-mediated transformation

of in vitro grown potato microtubers. Plant Cell Reports, v. 12, n. 6, p. 324-327, 1993.
https://doi.org/10.1007/BF00237428

TORRES, A. C.; TEIXEIRA, S. P.; POZZER, L. Cultura de ápices caulinares e recuperação de plantas livres
de vírus. In: TORRES, A. C.; CALDAS, L. S.; BUSO, J. A. (Eds). Cultura de tecidos e transformação
genética de plantas. Brasília: Embrapa-SPI, 1998. v. 1, p. 133-145.

TOVAR, P.; ESTRADA, R.; SCHILDERENTSCHLER, L.; DODDS, J. H. Inducción y utilización de

tubérculos in vitro de papa. Lima: Circular Centro Internacional de la Papa, 1985. 4p.

TOWILL, L. E. Survival at ultra-low temperatures of shoot tips from Solanum tuberosum group andigena,
phureja, stenotomum, tuberosum, and other tuber-bearing Solanum species. CryoLetters, v. 5, p. 319-326,
1984.

VARGAS, E. T.; GARCIA, E.; OROPEZA, M. Somatic embryogenesis in Solanum tuberosum from cell
suspension cultures: histological analysis and extracellular protein patterns. Journal of Plant Physiology, v.
162, p. 449-456, 2005. https://doi.org/10.1016/j.jplph.2004.07.001

VARGAS, T. E.; XENA, N.; VIDAL, M. C.; OROPEZA, M.; GARCIA, E. Genetic stability of Solanum
tuberosum L. cv. Desiree plantlets obtained from embryogenic cell suspension cultures. Interciencia, v. 33, p.
213-218, 2008.

VENKATASALAM, E. P.; SOOD, R.; PANDEY, K. K.; THAKUR, V.; SHARMA, A. K.; SINGH, B. P.
Development of low cost technology for in vitro mass multiplication of potato (Solanum tuberosum L.).
African Journal of Agricultural Research, v. 8, n. 49, p. 6375-6382, 2013. https://doi.org/10.5958/0974-
0112.2015.00069.9

VENKATASALAM, E. P.; SHARMA, S.; PANDEY, K. K.; SHARMA, S.; THAKUR, V.; SOOD, R.;
SINGH, B. P. Influence of hermetic and non-hermetic culture rooms on biochemical and in vitro morphological
characters of potato cultivars. Indian Journal of Horticulture, v. 72, n. 3, p. 353-358, 2015.

VIRMOND, E. P.; KAWAKAMI, J.; DIAS, J. A. C. S. Produção de minitubérculos de batata semente através
do aproveitamento de brotos. Horticultura Brasileira, v. 29, n. 2, p. 2819-2825, 2011.

WANG, P.; HU, C. In vitro mass tuberization and virus free seed-potato production in Taiwan. American
Journal of Potato Research, v. 59, n. 1, p. 33-37, 1982. https://doi.org/10.1007/BF02854881

WANG, K.; HE, W.; AI, Y.; HU, J.; XIE, K.; TANG, M.; WANG, Y.; VANDER ZAAG, P. Optimizing seed
potato production by aeroponics in China. Philippine Journal of Crop Science, v. 42, n. 1, p. 69-74, 2017.
https://doi.org/10.1007/s11540-006-9011-4

WANG, Q.; LIU, Y.; XIE, Y.;YOU, M. Cryotherapy of potato shoot tips for efficient elimination of Potato
leafroll virus (PLRV) and Potato virus Y (PVY). Potato Research, v. 49, n. 2, p. 119-129, 2006.

Biosci. J., Uberlândia, v. 34, n. 4, p. 952-969, July/Aug. 2018

 969
Application of tissue… MORAIS, T. P. et al.

WESTCOTT, R. Tissue culture storage of potato germplasm. 1. Minimal growth storage. Potato Research, v.
24, n. 3, p. 331-342, 1981a. https://doi.org/10.1007/BF02360370
https://doi.org/10.1007/BF02360371

WESTCOTT, R. Tissue culture storage of potato germplasm. 2. Use of growth retardants. Potato Research, v.
24, n. 3, p. 343-352, 1981b. https://doi.org/10.1007/BF02360370
https://doi.org/10.1007/BF02360371

WITHERS, L. A.; ENGELMANN, F. In vitro conservation of plant genetic resources. In.: ALTMAN, A.;
COLWELL, R. R. (Eds). Agricultural biotechnology. New York: Marcel Dekker, 1998. p. 57-88.

YAMAMOTO, S.; RAFIQUE, T.; FUKUI, K.; CASTILLO MARTINEZ, C. R.; NIINO, T. Application of V-
Cryo-plate method to in vitro shoot tips of potato varieties. In: INTERNATIONAL SYMPOSIUM ON PLANT
CRYOPRESERVATION, 2., 2013, Fort Collins, Colorado, USA. Anais… Fort Collins, 2013. p. 31.

YOON, J. W.; KIM, H.; KO, H.; HWANG, H.; HONG, E.; CHO, E.; ENGELMANN, F. Cryopreservation of
cultivated and wild potato varieties: effect of subculture of mother-plants and of preculture of shoot tips.
CryoLetters, v. 27, p. 211-222, 2006.

ZHANG, Z. J.; ZHOU, W. J.; LI, H. Z.; ZHANG, G. Q.; SUBRAHMANIYAN, K.; YU, J. Q. Effect of
jasmonic acid on in vitro explant growth and microtuberization in potato. Biologia Plantarum, v. 50, n. 3, p.
453-456, 2006. https://doi.org/10.1007/s10535-006-0069-2

ZOBAYED, S. M. A.; ARMSTRONG, J.; ARMSTRONG, W. Micropropagation of potato: evaluation of

closed, diffusive and forced ventilation on growth and tuberization. Annals of Botany, v. 87, p. 53-59, 2001.
https://doi.org/10.1006/anbo.2000.1299

Biosci. J., Uberlândia, v. 34, n. 4, p. 952-969, July/Aug. 2018