Anna Berlin, Yue Jin, ZS Kang, Kumarse Nazari, Ekaterina Skolo

1. The Barberry Connection:
Looking for the source of pathogen
variability in stem and stripe rusts
Panel discussion, BGRI Technical Workshop
Monday, September 3, 2012 (9:00-10:30 AM)

4. Barberry eradication laws
1600’s Ronen, France
1726-1766 New England colonies
CT, MA, RI
1800’s Many European countries
Denmark, England, France,
Germany, etc.
Stakman EC (1918) “The Black Stem Rust and the Barberry.” Yearbook of USDA – 1918, pages 75-100.

5. Barberry eradication, USA
1918-1975

6. >100,000,000
barberry bushes
destroyed
1. Delayed onset of disease
2. Reduced virulent inoculum in the spring
3. Decreased number of pathogenic races
4. Stabilized pathogenic races (e.g. QCC)
Durability of deployed genetic resistance
Roelfs AP (1982) Plant Disease 66:177-181

7. Known global hot-spots for the recent emergence of new races:
Areas without a history of systematic alternate host surveillance and control
Endemic barberries are found on every continent, except Australia
Nearly 500 barberry species, with high diversity in South America, Africa, and Asia

8. 1. What is the distribution of alternate hosts, relative to wheat production?
2. What role, if any, do these species play in rust epidemiology today?
A. Berlin, Sweden
E. Skolotneva, Russian Fed.
Y. Jin, USA
K. Nazari, ICARDA ZS Kang, PR China
W. Getaneh, Ethiopia
R. Wanyera, Kenya
Do alternate hosts undermine our efforts to achieve durable resistance?

9. THE BARBERRY STORY IN KENYA
Ruth Wanyera
Kenya Agricultural Research Institute (KARI), Njoro
Email:wanyera@plantprotection.co.ke

10. UG99 PRESENT STATUS
The rapid emergence of
Ug99 derivative races (a
"stacking" of virulences
first Sr31, then Sr24,
then Sr36, etc.)
suggests that sexual
recombination may play
a role in the evolution
of Pgt virulence in East
Africa
TTKSF

11. 1. Are there barberries in East Africa?
2. Are they susceptible to cereal rusts?
3. Do they function as alternate hosts under natural conditions?
LLL
Life cycle of stem rust

12. Records at the Kenya National museum
showed the presence of Berberis holstii
Engl., in East Africa

13. Barberry hunting and aecia collection since 2008

14. Barberry hunting…
Njoro Area
Mau-Narok
Mt. Kenya
Olkalau/Nyahururu
Narok
Meru (Chogoria Forest)
Mt. Elgon (Kenya Side)
Barberry surveyed sites
• Aecia positively identified in the lab

15. The barberry plant is
medicinal, fruits are
eaten by children
Dug up barberry bush
Locating barberry stands
required coordination of
the local communities
and experts outside the
wheat rust community

16. Inoculations
Inoculation process initiated in the
greenhouse at Njoro to determine if the
aeciospores produced on the barberry
leaves are wheat rusts, proved
unsuccessful (environmental conditions?).
KARI Muguga South (away from wheat
growing areas and barberry sites), but no
sporulation was observed ( unfavorable
conditions?)

17. Inoculations
 Efforts made trying to get aeciospores from Kenya
to the CDL for analysis, proved unsuccessful
because of viability issues

18. Way forward
 Need resources to conduct molecular diagnostics
locally
 In country capacity building to isolate and identify
Pgt from alternate hosts, due to fragility of
aeciospores relative to urediniospores

19. THANK YOU

20. Aecial Infections on Barberry Plants
(Zinkila) in Ethiopia
W. Getaneh, EIAR - Ambo
Beijing, China
September 2012

21. Introduction
• Around 1958, barberry (B. holstii) plant
surveys were conducted by the Biology
Department of Addis Ababa University in the
northern part of Ethiopia.
• In 1978, the Plant Protection Research Center
(PRCC) conducted barberry surveys in the
north Shewa zone; collected aeciospores were
inoculated on wheat seedlings, but no
infection was observed.

22. Introduction
In 2009, the pathology section of the PPRC
conducted B. holstii surveys to investigate
whether the plant functions as an alternate host
to stem rust in Ethiopia.
These surveys were carried out in the northern
part of Ethiopia (Shewa zone).

23. Surveys

24. Surveys
• In the north Shewa zone, 5 locations were
identified where B. holstii grows
• These locations range in altitude from 2781-
2895 m
• Abundant aeciospores were found on
barberry leaves from September to January.

25. Aeciospores on B. holstii leaves

26. Greenhouse Inoculation
Collected aeciospores were inoculated on the following species
identification set:
Genotype Species
McNair 701 Wheat
Line E Wheat
Morocco Wheat
Lemhi Wheat
Sr31/6*LMPG Wheat
Prolific Rye
Winter rye Rye
Hiproly Barley
Hypana Barley
Otana Oat

27. Greenhouse Inoculation

28. Greenhouse Inoculation

29. Greenhouse Inoculation

30. Greenhouse Inoculation

31. Inoculation results
From 6 inoculations, we obtained:
Infection
Genotype Species (# pustules)
McNair 701 Wheat 0
Line E Wheat 1
Morocco Wheat 0
Lemhi Wheat 0
Sr31/6*LMPG Wheat few
* Prolific
Winter rye
Rye
Rye
many
many
Hiproly Barley many
Hypana Barley many
Otana Oat 0
* Confirmed in US lab by both inoculation and DNA analysis

32. Conclusion
Our results suggest that B. holstii functions as an
alternate host to stem rusts of cereals in
Ethiopia, but this result is preliminary and will
require confirmation and further study.

33. Acknowledgements
I wish to acknowledge BGRI/DRRW project for financing
this activity and giving me the opportunity to participate
in this Technical Workshop.
Inoculation materials and research support were
generously provided by CDL.

34. Surveying aecial infections on Berberis spp.
in Central Russia
Ekaterina S. Skolotneva
Moscow Lomonosov State University
All-Russian Research Institute of
Phytopathology

35. Barberry survey in the Central Region of Russia (2000-2009)
. Sampling locations
... .
... ..
.. ..
.

36. • Are there any Berberis spp. in the
region?
• Yes.
B. vulgaris
B. purpurea

37. Barberry
bushes
Field of wheat crops
Golicyno-city
Barberry
bushes B. vulgaris
B. purpurea

38. Dates of disease development in Central Russia
May
Other
months
June
August July B. vulgaris
August, 2012: winter wheat
Nemchinovskaya 24 Agropyron repens

39. Sampling strategy from barberries
Location of
bushes = • We consider a location of bushes as a
sample
Collect fresh aeciospores sample. It may be single bush or
Inoculate suscep ble lines group of bushes which are the same
species and next to each other
oat rye wheat
• Only fresh aeciospores are collected
Score infected plants for evidence of infection
for inoculation
• From aecial samples we isolate
Susceptible Susceptible Susceptible different formae specialis of Puccinia
graminis:
Oat stem rust Wheat stem rust
is present Rye stem rust
is present
is present P. graminis f. sp. secalis
P. graminis f. sp. avenae
P. graminis f. sp. tritici
Susceptible alternate hosts to stem rust: B. vulgaris and B. purpurea.

40. Identification of Pgt races
• The International Set of 20 wheat differential lines was
used (upgraded in 2006, Prof. Yue Jin, USDA-ARS Cereal
Disease Laboratory, University of Minnesota, USA)
• From 2000 to 2009 twenty different Pgt races were
isolated from barberry
B. vulgaris
Year Barberry
2000 MKBT, MKLT, MKBP, TKNT, TKLT, RKNT
2001 MKBT, RKDT, PKJG, FKNL, TKNP, TKNT, TKFT
2002 MKBT, MKLT, MKBP, TKNT,
2003 MKNS, TTNT, TKNT, PKNT
2004 TKNT, TTNT,TKST, PKST
2005 TKNT, TTNT, TKST
2006 TKNT, TKPT, TKST, KJNT, RKNT, RKNS
2007 TKNTF, TKSTF
2008 TKNTF, TKNTC, TKSTF
2009 TKNTF, TKNTC, TKSTF

41. The frequencies of Pgt races isolated from barberry
120
MKBT MKLT TKNP TKFT
MKBP TKNT(F/C) TTNT TKST
100 TKPT TKNS TTNS PKST
PKNT
80
Race frequency
60
40
20
0
2000 2001 2002 2003 2004 2005 2006 2007 2008 2009
Years

42. Summary
Barberry species distributed in the Central region of Russia were
shown to serve as alternate hosts for stem rust pathogens under
natural conditions
A diversity of stem rust races was isolated from these species.

43. Alternate hosts panel
discussion
-P. graminis in the presence of
barberry
Anna Berlin

44. Reported
barberry
Background &
2000-
2012
law of barberry
eradication
• The Swedish law of barberry
eradication was repealed in 1994 Stockholm
• There is no formal survey regarding
presence of barberry
100km
www.artportalen.se

45. Harvest planting
fall-sown
cereals
Harvest
planting spring cereals
Jan Feb Mar Apr May Jun Jul Aug Sept Oct Nov Dec

46. Markers enable connecting
populations from the two hosts
K=6 data from one structure run
Berlin et al. 2012 Phytopathology

47. X X X
X X X
Diversity X X X
X X X
• 30 samples = 30 genotypes
X X X
• 30 genotypes = ? Races
100 m
X X X
X X X
X X X
X X X
X X X
30 m

48. Why no stem rust on wheat?
• Barberry present
• Severe epidemics on oats and natural grasses
• Some rust on rye
• Large genotypic variation within and between fields
• P. graminis clearly completing its sexual cycle
• Does the population identified as P. graminis f. sp.
tritici/secalis lack the virulence necessary to infect
wheat?
• Does wheat grown in Sweden have effective
resistance genes?

50. Research progress on alternate host
and sexual stage of wheat stripe rust in China
Zhensheng Kang
State Key Laboratory of Crop Stress Biology for Arid Areas,
Northwest A & F University,Yangling, Shaanxi, China

51. Wheat stripe rust (Pst) is a destructive disease
throughout all winter wheat regions in China and is
considered the most important disease of wheat.
 Diseased area: between 3-6 million ha.
 Yield losses: 10-50%.
Years Losses
(million tonnes)
1950 6.0
1964 3.2
1990 2.6
2002 1.4

52. Based on historical epidemiological data for stripe rust,
the wheat-growing regions in China can be divided into
three areas:
 Western over-summering areas
 Over-wintering areas
 Eastern epidemic areas
Over-summering areas
Over-wintering areas
Eastern epidemic areas

53. In the mountainous western areas, stripe rust can
over-summer on volunteer wheat and late-maturing
spring wheat.
Over-summering areas in south Guansu Volunteer wheat in south Guansu

54. Hebei
Shanxi Shandong
Gansu Henan
Jiansu
Shaanxi
Anhui
Hubei
Sichuan Chongqing
oversummer
The migration pathway of P. striiformis in China. movement

55. The western over-summering areas are
considered a “hot-spot” for the emergence of
new races of wheat stripe rust in China.
 Most new races were first detected in these regions
in recent history;
 A high genetic diversity within the regions’ Pst
populations has been reported by different research
groups (Lu et al., 2011; Duan et al., 2010; Mboup
et al., 2009);
 The genetic recombination was found to occur for Pst
in these regions (Lu et al.,2011; Duan et al., 2010;
Mboup et al., 2009).

56. Why does the western over-summering areas
become the “hot-spot” for wheat stripe rust
in China?
Virulence variation for rusts maybe due to
 sexual hybridization
 mutations
 somatic hybridization
However, the mechanism of sexual hybridization
for wheat stripe rust has been neglected since
the sexual stage was presumed to be absent.

57. In 2010, some Berberis spp. were shown to serve as
alternate hosts for the wheat stripe rust pathogen.
 B. chinensis,
 B. holstii,
 B. koreana
 B. vulgaris.

58. Questions: Does the sexual stage of
wheat stripe rust occur under natural
conditions, particularly in China?
 Do any susceptible barberry species coexist
with wheat in China?
 Can wheat stripe rust be isolated from
infected Berberis spp. in China?
 Does the sexual stage of wheat stripe rust
contribute to variation in virulence?

59. Surveys for Berberis spp.in China
 215 of the ~500 described Berberis spp. in the
world are endemic to China;
 Many of China's Berberis spp. are distributed in the
western over-summering areas (hot-spot).
Distribution of Berberis species in different regions of China
Western China:
Sichuan：81; Chongqin： 30; Yunnan： 78; Tibet: 55; Guansu： 26;
Shaanxi: 20; Guizhou： 19; Qinghai：13; Xinjiang： 5; Ningxia： 3;
Central China:
Hubei： 24; Henan： 7; Shanxi： 10; Hunan： 9; Anhui： 2;
Eastern China:
Hebei： 6; Jiangxi： 5; Guandong：4; Guanxi：4; Hujian： 5;

60. B. shensiana B. brachypoda
Berberis soulieana B. potaninii

61. Are these Berberis spp.
susceptible to wheat
stripe rust ?

62. Identification of Berberis spp. as alternate
hosts of wheat stripe rust
Dew chamber
We collected seeds and seedlings of Berberis spp. from the field
and inoculated using telia of Pst in the greenhouse.

63. Identification of Berberis spp.
as alternate hosts of wheat stripe rust
Pycnia on Berberis
Infection of basidiospore and development of pycnia
Normally, we see pycnia on the leaves of susceptible
Berberis spp. 11-14 days after inoculation.

64. Identification of Berberis spp.
as alternate hosts of wheat stripe rust
About 20 days after inoculation, we see aecia develop on the leaves.
Aeciospores can infect wheat through the stoma and produce typical
rust symptoms (uredinia).

65. Species of barberry identified as alternate hosts of Pst by
artificial inoculation, using germinating teliospores in China
No. Berberis spp. Orgin Distribution
1 B. aggregata Gansu, China Gansu, Sichuan, Hubei, Qinghai, Shanxi
2 B. brachypoda Gansu, China Gansu, Sichuan, Hubei, Qinghai, Shanxi,Henan, Shanxi
3 B. potaninii Gansu, China Gansu, Shaanxi, Sichuan
4 B. soulieana Gansu, China Gansu, Shaanxi, Sichuan,Hubei
5 B. dasystachya Shaanxi, China Gansu, Shaanxi,Hubei,Shanxi
6 B. shensiana Shaanxi, China Shaanxi,Gansu
7 B. atrocarpa Sichuan, China Sichuan, Yunnan, Hunan
8 B．ferdinandi-coburgii Yunnan, China Yunnan
9 B. phanera Yunnan, China Yunna, Sichuan
10 B. aggregate var. Yunnan, China Gansu, Sichuan,Qinghai,Hubei,Shanxi
integrifolia
11 B. davidii Yunnan, China Yunnan
12 B. stenostachya Gansu, China Gansu, Shaanxi, Shanxi
13 B. wangii Yunnan, China Yunnan
14 B. circumserrata Shaanxi, China Shaanxi, Hubei, Gansu, Qinghai,Henan
15 B. poiretii Beijing, China Shaanxi, Qinghai, shanxi, Hebei, Jilin, Liaoning,
16 B. guizhouensis Guizhou, China Guizhou

66. Berberis soulieana
B. shensiana
 Some species are evergreen, distributed in southwest regions.
 Others are deciduous, distributed in northwest regions.
 Some susceptible species (e.g., Berberis soulieana, B. brachypoda,
and B. shensiana) are widely distributed in the western over-
summering areas.

67. Are any Berberis spp. infected by wheat stripe
rust under natural conditions in China?
Aecia produced on barberry leaves in nature
We collected 3703 infected- barberry leaves in the fields
and inoculated wheat with aeciospores in the greenhouse.

68. Are any Berberis spp. infected by wheat stripe
rust under natural conditions in China?
Berberis brachypoda B2011-1 B2011-2 B. Shensiana B2011-3 B. Soulieana B2011-4
Four stripe rust cultures (B2011-1, B2011-2, B2011-3, and B2011-4) from three barberry (Berberis
spp.) species including B. brachypoda , B. soulieana , and B. shensiana collected from Gansu and
Shaanxi Provinces in 2011, respectively.
Berberis species Origin Number of Number of uredium-culture
aecium-isolate produced on wheat Mingxian 169
Berberis Gansu 1519 2
brachypoda
B. shensiana Shaanxi 410 1
B. soulieana Gansu 384 1
B. potaninii Shaanxi 742 0
B. aggregate Gansu 648 0

69. Can any Berberis spp. be infected by wheat
stripe rust under natural conditions in China?
A B
M M M
Ethidium bromide(EB) stained agarose gel showing that four isolates B2011-1, 2011-2,2011-3, and 2011-4 from naturally rust-infected barberry
species produced uniform single band amplified using primers ITS-puccinia (5'-ACATCGATGAAGAACACAGT-3' )/ITS4( 5'-
TCCTCCGCTTATTG-ATATGC-3')(left, part A), and specific-primers PSF(5'-GGATGTTGAGT-GCTGCTGTAA-3' )/PSR (5‘-
TTGAGGTCTTAAGGTTAAAA-TTG-3' ) (right, part B) in accord with race CYR 32 of Puccinia striiformis f. sp. tritici as positive control in
size. Sterile water used as negative control (NC). M= 100bp DNA marker DL2000.
PCR tests using two Pst-specific primer pairs demonstrate
the recovery of four cultures of Pst from Berberis spp.

70. Comparison of infection types on Chinese differential hosts of
eight major races of Pst and the four Pst cultures recovered
from three barberry species
Cultures Differential hosts
and Origin of cultures
races 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
B2011-1 Berberis brachypoda R S R S S S R S R SR S R R S R R R R R
B2011-2 B. brachypoda R S R S S R R S R R S R R R R R R R R
B2011-3 B. shensiana R S R S S R S S R RS S R R RS R R R R R
B2011-4 B. soulieana R S R R S S R S R R R R R S R S R R R
CYR33 Triticum aestivum S S S S S S S S S S S S S S R S R R R
CYR32 T. aestivum S S S S S S S S S S S S S S R S S R R
CYR31 T. aestivum S S S S S S S S S R S S R S R S S R R
CYR30 T. aestivum S S S S S S S S S R S S R R R S S R R
CYR29 T. aestivum S S S S S S S S S R S S R R R S R R R
CYR28 T. aestivum S S S S S S S S S R S R R R R S R R R
CYR23 T. aestivum S S S S R S S S S R S R R R R R R R R
CYR17 T. aestivum SR S R SR R SR S R R R RS R R R R R R R R
Virulence tests demonstrated that the infection types of
the four barberry-derived cultures are different compared
to the major Chinese races.

71. Does the sexual stage under natural
conditions contribute to variation in
virulence for wheat stripe rust in China
→ → →
Aecia
from Berberry
Recovered culture
from aecium
Single-uredium Virulence test
isolates on differentials

72. Virulence difference among single-uredium isolates from
single-aecium-derived culture (B2011-2) on differentials
Infection type of single-uredium isolates on differential hosts
No. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
B2011-2 -1 R S R S R R R S R R S R R R R R R R R
B2011-2 -2 R S R R R R R S R R R R R S R R R R R
B2011-2 -3 R S R S R R R S R R S R R R R R R R R
B2011-2 -4 R S R S R R R S R R S R R S R R R R R
B2011-2 -5 R S R R R R R S R R S R R R R R R R R
B2011-2 -6 R S R S R R R S R R R R R R R R R R R
B2011-2 -7 R S R S S R R S R R S R R R R R R R R
The virulence tests showed high diversity in virulence
among the single-uredium isolates.

73. Virulence difference among single-uredium isolates from single-
aecium-derived culture (B2011-1) on differentials
No. Infection type of single-uredium isolates on differential hosts
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
B2011-1-1 RS S R S R S RS S R R R R R S R S R R R
B2011-1-2 R S R S S S R S R R R R R S R R R R R
B2011-1-3 R R R S S S RS S R RS RS R R S R R R R R
B2011-1-4 R S R S S R R S R S R R R S R R R R R
B2011-1-5 R S R R R S SR SR R R SR R R S R R R R R
B2011-1-6 R S S S R S R S R R S R R S R R R R R
B2011-1-7 R S S S S R R S R S S R R R R R R R R
B2011-1-8 R S R SR SR S RS S R R RS SR R S R R R RS R
B2011-1-9 R S S S S S R S R R RS R R S R R R R R
B2011-1-10 R S R S SR S R S R SR SR R R S R R R R RS
B2011-1-11 R S S S R S R S R S R R R S R R R R R
B2011-1-12 R S R S SR SR R S R R S R R RS R R R R R
B2011-1-13 R S S S S S R S R S R R R S R R R R R
B2011-1-14 R S R R R S RS S R RS R R R R R R R R R
B2011-1-15 R S R S SR S R S R R RS RS R S R R R R R
B2011-1-16 R S R S R S R S R RS RS RS R S R RS R R R
B2011-1-17 R S S S S S R S R S R R R S R R R R R
B2011-1-18 R S R S S S RS S R SR R R R S R R RS R R
B2011-1-19 R S R R RS S R S R R RS R R R R R R R R
B2011-1-20 R S R S S S R S R SR RS R R RS R R R R R
B2011-1-21 R S R S S S R S R SR S R R S R R R R R
B2011-1-22 R S R S S S R S R SR S R R S R R R R R

74. Summary
 So far, 16 of the 22 tested barberry species from over-
summering areas show susceptibility to wheat stripe rust,
indicating that there is a great diversity of potential alternate
hosts for stripe rust in China.
 Four cultures of wheat stripe rust were obtained from three
barberry species in the field, indicating that the sexual stage
of wheat stripe rust occurs under natural conditions in China.
 Virulence tests for single-uredium isolates recovered from a
single aecium demonstrate that the sexual stage contributes to
variation in virulence for the wheat stripe rust pathogen in
China.

76. Future Work
 More barberry species need to be tested for their
susceptibility to Pst in China;
 More field Surveys need to be conducted;
 More evidence is needed to elucidate the
relationship between genetic diversity and the
sexual stage of Pst,

77. Acknowledgement
 The earmarked fund for Dr. Jie Zhao, Prof. Lili Huang,
Modern Agro-industry Dr. Hongchang Zhang, Dr. Dejun Han,
Dr. Xiaojie Wang, Dr. Chunfang Wang,
Technology Research System in
Dr. Qingmei Han, Dr. Jun Gou,
China
Mrs. Guorong Wei, Dr. Xueling Huang,
 National Basic Research Dr. Gang Zhang, Dr. Yonghong Zhang,
Program of China (973) Dr. Xiumei Yu, Dr. Changqing Chen,
 Nature Science Foundation of Dr. Liangsheng Xu, Dr. Ninghai Lu,
China Dr. Bo Liu, Dr. Jingbiao Ma,
 The 111 Project from the Mr. Gangming Zhan, Dr. Wenming zheng
Chinese Ministry of Education All Ph D and Master studendts in My Lab.
Dr. X M. Chen, Dr. H. Buchenauer,
Dr. Robert McIntosh , Dr. Colin
Wellings, Dr. Scot H. Hulbert,
Dr. Jin-Rong Xu, Dr. Shiping Wang,
Dr. Hei Leung, Dr. J. Chong
Dr. Yue Jin, Dr. Ravi Singh,
Dr. Zacharias Pretorius

78. Thanks for your attention!

80. Berberis rust survey in the Ug99 pathway in CWANA
Kumarse Nazari ICARDA
Annemarie F. Justesen GRRC, Aarhus University
Jens Grønbech Hansen GRRC, Aarhus University
Dave Hodson CIMMYT
Mehran Patpour, Farzad Afshari, Seed and Plant Improvement Institute, Karaj Iran
Hojjatollah Rabbani Nasab North Khorassan Agricultural and Natural Resource
Zoia Sikharulidze Institute of Plant Immunity from Georgia
Amir Amanov, Zafar Ziyaev Kashkadarya Grain Breeding and Seed Production
Institute, Uzbekistan
Atiq ur Rehman Rattu National Agricultural Research Centre, Pakistan
Hukmatullo Ahmadov, Mahbubjon Rahmatovm, Bahiram Tajik Academy of Agricultural Sciences
Konul Aslanova Agrarian and Animal Husbandry Research Institute,
Azerbaijan

81. Methodology
•Berberis survey and biological assays
•Photo documentation of the Berberis host
•Leaf sample collection of Berberis spp.
•DNA-extraction from single aecial pustules,
several pustules from each barberry plant
•PCR amplification of EF1α- and/or β-tub-gene
•Species identification by sequence comparisons to
sequences in GenBank and reference sequences
from grass and cereal hosts

82. DNA sequence data obtained so far:
•Iran: 12 sequences (2010), 4 (2012)
•Tajikistan : 24 sequences (2011)
•Uzbekistan: 7 sequences (2012)
•Azerbaijan : 6 sequences (2012)

85. Kelardasht N 36.5002- E 51.1683- H 1224
Z.STO Z.KELMP- TP 1 ZERESHK ZER89 TP3- ZERESHK 89 ZERESHK 89-
No. Name /Line Gene ZER 88
L SP. KEL89 3 TP3 TP2
1 ISr5-Ra Sr5 33+ 3 4 4 4 33+ 4
2 CnS_T_mono_deriv Sr21 33+ 3 4 4 4 12+ 4
3 Vernsatine Sr9e 2+3 3 4 4 4 3+ 4
4 ISr7b-Ra Sr7b 4 3 4 4 4 3+ 4
5 ISr11-Ra Sr11 4 4 4 4 ;1 4 ;C11-
6 ISr6-Ra Sr6 4 4 4 4 4 3+ 33+
7 ISr8a-Ra Sr8a 4 4 4 4 4 3+ 33+
8 CnSr9g Sr9g 4 3+ 4 4 4 33+ 3+
9 W2691SrTt-1 Sr36 4 3 4 4 33+ 33+ 4
10 W2691Sr9b Sr9b 4 33+ 4 4 4 3+ 4
11 BtSr30Wst Sr30 4 33+ 4 4 4 3+ 4
12 Combination VII Sr17 3+ ;1 4 3+ 4 3+ 4
13 ISr9a-Ra Sr9a 4 3+ 4 4 4 3/33+ 3+
14 ISr9d-Ra Sr9d 4 3 4 4 4 3/33+ 3+
15 W2691Sr10 Sr10 4 3 4 4 4 3+ 3+
16 CnsSrTmp SrTmp 33+ 2- 4 4 4 ;C1= 3+
17 LcSr24Ag Sr24 ;1 ;1- 11-C 1+2-C ; ;C 1-
18 Sr31/6*LMPG Sr31 ; 3+ 0;1= 1C ;1 ;C1- ;C1=
19 Trident Sr38 X+ 4 ;C1= 3+ 3x X- ;;CN
20 McNair 701 SrMcN 4 4 4 4 4 3+ 33+
Race Identified TTTTF TTSSK TTTTC TTTTF TKTTF PTTSC TKTTC

86. Preliminary results
• Two main clusters: P. graminis and P. striiformis
•Within the P. graminis cluster, sequences show 94-95%
identity to Pgt sequences in GenBank
•All sequences within the P. striiformis cluster are from
Tajikistan and show 99-100% identity to P. striiformis
f.sp. dactylis (P. striiformoides), none are identical to P.
striiformis f.sp. tritici

87. Future work
Continue sequencing of more aecia
Sequence β-tubulin or ITS in order to be able to identify host
origin based on sequences in GenBank
Obtain more reference sequences from grasses??

89. Looking for the Source of Pathogen Variability in Stem and
Stripe Rusts—the Barberry Connection
--- Knowledge Gaps and Challenges
Yue Jin
USDA-ARS Cereal Disease Laboratory
University of Minnesota, St Paul, Minnesota, USA

90. “Cereal rusts are the most-researched plant diseases”
“A large body of literature exists”

91. “Cereal rusts are the most-researched plant diseases”
“A large body of literature exists”
Interpretation: there are no more mysteries!

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Puccinia graminis from Mahonia spp.

93. “Does barberry play a role in pathogen variation and disease
epidemiology in stem rust and stripe rust?”
e.g. the Kenya example:
Are there barberries?
----- Yes (B. holstii)
Is B. holstii susceptible to stem rust?
----- Yes
Does B. holstii function as an alternate host in Kenya?
----- Do not know

94. “What is the species?”
Is there any taxonomic support when we encounter unknown species?
Can we develop a robust assay to identify Berberis spp.?

95. Barberry is back.

96. Please remain skeptical!

97. The Barberry Connection:
Looking for the source of pathogen
variability in stem and stripe rusts

98. Sr31 virulence
Sr31 + Sr24 (Kenya)
detected in Uganda
(Ug99) Sr31 + Sr36 (Kenya)
2000 2004 2008 2012
Ethiopia Iran S. Africa Eritrea Ug99
Uganda Kenya Sudan Tanzania
Yemen Zimbabwe races
Sounding the Alarm
Global Rust Initiative (GRI)
1. Race surveillance
2. Resistance screening
3. Breeding
4. Chemical control
5. Seed production
6. Impact assessment
7. Training
8. Infrastructure (E. Africa)
9. Reporting and communication
10. Resources for IARCs

99. Sr31 virulence
Sr31 + Sr24 (Kenya)
detected in Uganda
(Ug99) Sr31 + Sr36 (Kenya)
2000 2004 2008 2012
Ethiopia Iran S. Africa Eritrea Ug99
Uganda Kenya Sudan Tanzania
Yemen Zimbabwe races
Sounding the Alarm Sexual populations of
wheat stripe rust found
in China
East African native
barberry (B. holstii) Barberry spp. shown
shown to be susceptible to function as alternate
to stem rust hosts to wheat stripe rust
Aecial infections on B. holstii shown to be
B. holstii found in susceptible to stripe rust
East Africa
Sexual populations of
wheat stem rust found on
Mahonia spp. in PNW
Sexual populations of
oat stem rust found
in Sweden

100. Training video: barberry.globalrust.org

101. The Barberry Connection:
Looking for the source of pathogen
variability in stem and stripe rusts
A Berlin, Sweden
Y Jin, USA
ZS Kang, China
K Nazari, ICARDA, Syria
E Skolotneva, Russian Federation
R Wanyera, KARI, Kenya
G Woldeab, EIAR, Ethiopia