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CURRENT APPLICATION OF ANTHER CULTURE AS A TOOL FOR

IMPROVEMENT OF HORTICULTURAL CROPS

Poster · May 2018

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3 authors, including:

Fidanka Trajkova Liljana Koleva Gudeva

Goce Delcev University of Štip Goce Delcev University of Štip
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 Current application of anther culture as a tool for
improvement of horticultural crops
Marija Pockovska, Fidanka Trajkova*, Liljana Koleva Gudeva
Department of Plant Biotechnology, Faculty of Agriculture
Goce Delcev University – Stip, Republic of Macedonia
*Corresponding author: fidanka.trajkova@ugd.edu.mk

Abstract Up-to-date research in androgenesis of pepper (Capsicum annuum L.)

Author Medium Starting explants Results
Anther culture is one of the plant biotechnology tools utilized for creation of haploid and spontaneous
dihaploid plants from different horticultural crops. Androgenesis can be defined as a set of biological Quin & Rotino, 1995 • C + 0.5 mg/l 2.4-D + 0.5 mg/l KIN Anthers Embryons
processes leading to an individual that genetically originated exclusively from an anther microspores that • C + 0.5 mg/l 2.4-D + 0.6 mg/l 6-BAP
represent the mail genotype. The main objective of in vitro anther culture is to obtain dihaploid Koleva-Gudeva & Spasenoski, • MS + 0.1 mg/l KIN + 0.01 mg/l 2.4-D + 0.001 Anthers Callus and
homozygous lines with high practical value for breeders, but researches in this area also have theoretical 2002 mg/l IAA embryons
importance for fundamental biological sciences. After spontaneous, or induced genome doubling, • N + 1.0 mg/l KIN + 0.001 mg/l IAA
dihaploids that are completely homozygous can be utilized differently in fundamental and practical • LS + 3.0 mg/l KIN + 1.0 mg/l IAA
research programs. From the standpoint of plant breeding, androgenesis has power to reduce the typical • NN + 0.01 mg/l KIN + 0.001 mg/l 2.4-D
7–9 inbreeding generations necessary to stabilize a hybrid genotype to only one. It is the key advantage of • CP + 0.001 mg/l KIN + 0.01 mg/l 2.4 – D
dihaploid technology in the context of plant breeding. R1 + 0.1 mg/l KIN
Since pepper, tomato and eggplant are one of the most important Solanaceae crops worldwide, the Koleva-Gudeva, 2003 • CP + 0.01 mg/l KIN + 0.01 mg/l 2.4-D Anthers Shoots
improvement of their diversity is possible by engagement of the methods of classical breeding, but also by R1 + 0.01 mg/l KIN
plant biotechnology which can advance the breeding process. During the past years, in vitro anther culture Nowazyk et al. 2006 • CP + 0.01 mg/l 2.4-D + 0.1 mg/l KIN + 0.5 g/l Anthers Embryons
research of numerous horticultural crops is extended although its successful application depends on activated carbon + 5 mg/l AgNO3 + 8 g /l agar
different factors. Hence, in vitro anther culture is one of the biotechnological tools which is often exploited
in different solanaceous and other horticultural species pre-breeding programs. Rodeva et al. 2007 • CP+0.01 mg/l KIN + 0.01 mg/l 2.4-D Anthers Shoots, embryos,
R1+0.01 mg/l KIN embryoid
• MS+ 0.3 mg/l 2.4-D + 0.1 mg/l KIN + 0.005 mg/l structures
Up-to-date research in androgenesis androgenesis of tomato (Lycopersicon esculentum Mill.) Biotin + 0.1 mg/l Glycine + 0.04 mg/l Vitamin
Author Medium Starting explants Results B12 + 30 g/l sucrose + 0.7% Agar

Özzambak, 1994
Zagorska et al., 1998
Segui-Simarro & Nuez, 2005
• Nitsch + 2 mg/l NAA + 1 mg/l K + 40 g/l sucrose Anthers Callus Koleva-Gudeva et al. 2007 • CP + 0.01 mg/l KIN + 0.01 mg/l 2.4-D Anthers Shoots and direct
• MS + 2 mg/l 2.4-D + 0.1 mg/l K + 30 g/l sucrose R1 + 0.01 mg/l KIN somatic embryoids
Koleva – Gudeva, 2008 • MS + 0.1 mg/l KIN + 0.01 mg/l 2.4-D + 0.001 Anthers Embryoids and
mg/l IAA callus
• MS basic + 1 mg/l 2 ip + 2 mg/l IAA Anthers Shoots • N + 1.0 mg/l KIN + 0.001 mg/l IAA
• MS basic + 0.25 mg/l zeatin + 0.5 mg/l IAA • LS + 3.0 mg/l KIN + 1.0 mg/l IAA
• NN + 0.01 mg/l KIN + 0.001 mg/l 2.4-D
• CP + 0.001 mg/l KIN + 0.01 mg/l 2.4 – D
For anthers: Anthers and From anthers:
R1 + 0.1 mg/l KIN
microspores Callus and shoots
• MS 2.5 g/l Phytagel + 20 g/l sucrose + 1 mg/l 2ip + 2 Koleva-Gudeva & Trajkova, • CP + 0.01 mg/l KIN + 0.01 mg/l 2.4-D Anthers Shoots
mg/l IAA From microspores: 2012 R1 + 0.01 mg/l KIN
• NLN medium + 2.5 g/l phytagel +130 g/l sucrose + Proembryos Koleva-Gudeva et al. 2013 • CP + 0.01 mg/l KIN + 0.01 mg/l 2.4-D Anthers Callus and
0.5 mg/l BAP + 0.5 mg/l NAA R1 + 0.01 mg/l KIN embryos

For microspores:

• MS and 20 g/l sucrose + 1 mg/l 2ip and 2 mg·1-1 IAA

B C
• NLN medium +130 g/l sucrose + 0.5 mg/l BAP + 0.5
mg/l NAA
A B
Azar 2010 • MS basic + 2 mg/l IAA + 1 mg/l 2ip + 20 g/l glucose + Anthers Shoots
D E
7 g/l agar A

Corral-Martinez et al., 2010
• DBM1 + 5 mg/l kin + 2 mg/l NAA + 20 g/l sucrose
• MS + vitamins + 2.5 g/l Phytagel + 20 g/l sucrose, 1
mg/l 2 ip and 2 mg/l IAA
А) Plants from different pepper genotypes used as anther
donors; B) Isolated pepper anthers dyed with aceto-carmine; C)
Anthers Shoots Mature pollen grains from pepper, dyed with aceto-carmine; D)
C D Microspores in stadium of first pollen division (х400); E)
Microspores after 6 days cultivation (х400)

Up-to-date research in androgenesis of eggplant androgenesis (Solanum ovigerum Dunal)

Author Medium Starting explants Results
E F
Özzambak, 1994 • Nitsch + 2 mg/l NAA + 1 mg/l K + 40 g/l sucrose Anthers Callus

Rizza et al., 2002 • C6 + 5 mg/l Kin + 5 mg/l NAA Anthers Embryos and callus
• C9 + 1 mg/l Zeatin + 3 mg/l NAA
• C12 + 0.5 mg/l TDZ + 0.1 mg/l Zeatin + 0.5 mg/l IAA Four-years experimental
design for agronomic Androgenic pepper genotypes
G H and their parental genotypes for
evaluation of three
Salas et al., 2011 • agar-based Ct inductive medium Anthers Embryos and callus А) and В) Anthers on induction medium С) Formation of androgenic pepper agronomic evaluation experiment
• R1 medium embryoid; D) Emergence of embryoid from anther after genotypes (Kurtivska kapija, set up in randomized block design
30 days cultivation Е) Androgenic pepper plants on V3 in the experimental greenhouse
Rivas-Sendra et al., 2017 • C medium + 120 g/l sucrose + 8 g/l Bacto-agar + 5 Anthers Shoots Piran and Fezerozon)
medium; F) Fully developed plants for acclimatization in
mg/l kinetin and 5 mg/l 2,4-D
climate chamber; G) Karyotype of control genotype
Kurtovska kapija; H) Karyotype of androgenic plant from
Androgenesis is one of the methods utilized for creation of haploid and spontaneous dihaploid Feherozon
plants in in vitro culture. Gained haploids and dihaploids have certain genetic potentials which
are quickly phenotypically expressed because they origin from a haploid cell. These androgenic
plants, before being included in breeding process, must be studied for their growth and
development stages, their genetic and morphological characteristics compared to the mother
genotype which give information about similarities and differences between androgenic and
mother genotypes. Fully regenerated androgenic plans from different pepper genotypes
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