Journal of Phytopathology. 2017;1–8. wileyonlinelibrary.com/journal/jph
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1 © 2017 Blackwell Verlag GmbH
Received: 8 February 2017
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Accepted: 10 April 2017
DOI: 10.1111/jph.12583
ORIGINAL ARTICLE
Assessment of genetic diversity of coffee leaf rust pathogen
Hemileia vastatrix using SRAP markers
Bharathi Kosaraju
1
| Soundararajan Sannasi
2
| Manoj Kumar Mishra
1
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Daivasikamani Subramani
3
| Muniswamy Bychappa
1
1
Plant Biotechnology Division, Unit of Central
Coffee Research Institute, Coffee Board,
Manasagangothri, Mysore, Karnataka, India
2
Regional Coffee Research Station,
Thandigudi, Dindigul, Tamil Nadu, India
3
Central Coffee Research Institute, Coffee
Research Station, Chikmagalur, Karnataka,
India
Correspondence
M. K. Mishra, Plant Biotechnology Division,
Unit of Central Coffee Research Institute,
Coffee Board, Manasagangothri, Mysore,
Karnataka, India.
Email: manojmishra.m@gmail.com
Abstract
Coffee leaf rust caused by the fungus Hemileia vastatrix (Berk and Br.) is a major dis-
ease occurring in coffee plantations. Although the rust fungus exists in different physi-
ological races, the genetic difference between them is meagrely understood. In this
study, genetic diversity of 14 identified and two unidentified leaf rust races was deter-
mined by sequence-related amplified polymorphism (SRAP) markers. Of 48 SRAP
primer pairs tested, 35 primers are polymorphic and generated 347 distinct scorable
fragments. The number of fragments ranged from 4 to 18 with a mean of 9.97 frag-
ments per primer combination. Of the total 347 amplified fragments, 185 fragments
(53.31%) are polymorphic with an average of 5.41 fragments per primer combination.
The average resolving power (Rp) and the average polymorphism information content
(PIC) of the 35 SRAP primer combinations were 13.60 and 0.356, respectively. Of 35
SRAP primer pairs, 15 primer pairs were more informative and generated 25 unique
fragments, which are useful for race discrimination. The study demonstrated the exist-
ence of genetic variability among various leaf rust races and this information will be
helpful in coffee breeding programmes.
KEYWORDS
coffee, genetic variability, Hemileia vastatrix, molecular markers, physiological races
1 | INTRODUCTION
Coffee is one of the most important crops in the tropics and considered
to be the second most important commodity in the world trade market
next to petroleum. The total global production of green coffee is ap-
proximately 143.25 million bags (60 kg capacity) with an export earning
of over US$ 31.8 billion during 2014. Although more than 100 coffee
species are known, only two species, that is, Coffea arabica (known as
arabica coffee) and Coffea canephora (known as robusta coffee), are
commercially cultivated and consumed. The economy of a number
of countries depends heavily on the earnings of this crop.Coffee leaf
rust (CLR), popularly known as orange rust, is caused by the biotrophic
fungus Hemileia vastatrix Berk. & Br. It is one of the major diseases in
coffee species reported in almost all coffee-growing countries of the
world. Although both C. arabica and C. canephora are affected by the
leaf rust disease, the production and productivity of arabica coffee are
severely determined by the incidence level. It has been estimated that
in severe incidence level, the yield reduced from 30% to 80% in the
case of arabica coffee. Although chemical control of CLR disease by
using different fungicides was largely practised in plantation, the use of
rust-tolerant coffee genotype was considered to be the most promising
alternative (Várzea & Marques, 2005; Zambolim, 2016). In India, coffee
breeding aimed for rust resistance has been taken up and continued
for the last 80 years, and as a result, many resistant cultivars have been
released for commercial cultivation. In the majority of these breeding
programmes, tetraploid interspecific hybrids such as Hybrido de Timor
(HDT), Devamachy (putative tetraploid hybrid between C. arabica and
C. liberica), Ligenioides (spontaneous tetraploid hybrid of C. liberica and
Coffea eugenoides) as well as the hybrids obtained from the cross be-
tween C. arabica and Coffea canephora are used as the donor parent for