Corporate Profile 47Billion Information Technology
2nd iwsrs kang(2014-4-29)
1. Barberry plays an important role in natural
infection of stripe rust on wheat in China
Zhensheng Kang
State Key Laboratory of Crop Stress Biology for Arid Areas,
Northwest A & F University, Yangling, Shaanxi, China
2. 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
Wheat stripe rust is a destructive disease
throughout all winter wheat regions and is
considered the most important disease of
wheat in China.
3. Virulence variation for rusts maybe due to
sexual hybridization
mutations
somatic hybridization
However, the mechanism of sexual hybridization
for Pst has been neglected since the sexual
stage was presumed to be absent.
Resistant
wheat
cultivars
Rust
virulence
variation
Huge
yield
losses
4. 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.
Jin et al. Phytopathology, 2010
5. Question: Does the sexual stage of wheat
stripe rust occur under natural
conditions, particularly in China?
Field Surveys for Berberis spp.
Susceptible test of Berberis to stripe rust
Isolation of stripe rust from nature-infected
Berberis
Virulence test for recovered isolates
Mock-inoculation on Berberis in fields
6. 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. distribute in the
western over-summering areas (hot-spot).
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:
Distribution of Berberis species in different regions of China
10. Identification of Berberis spp. as alternate
hosts of wheat stripe rust in green house
Dew chamber
We collected seeds and seedlings of Berberis spp. from the
fields and inoculated using telia of Pst in the greenhouse.
11. Normally, we see pycnia on the leaves of susceptible
Berberis spp. 10-13 days after inoculation.
Infection of basidiospore and development of pycnia
Pycnia on Berberis
Identification of Berberis spp. as alternate
hosts of wheat stripe rust in green house
12. 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).
Identification of Berberis spp. as alternate
hosts of wheat stripe rust in green house
13. 23 Berberis species: alternate host for Pst
Successful production of pycnia, and aecia on
upper and back sides of barberry leaf after inoculation
Zhao, J., et al. Phytopathology, 2013, 103(9):927-934
14. A B C D E F
G H I J K L
M N O P Q R
Zhao, J., et al. Phytopathology, 2013, 103(9):927-934
15. 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. integrifolia Yunnan, China Gansu, Sichuan,Qinghai,Hubei,Shanxi
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
17 B. wilsonae Yunnan, China Yunnan, Tibet, Shaanxi, Qinghai, Hubei, Guizhou
18 B. sp. (unidentified) Tibet, China Tibet
19 B. jamesiana Yunnan, China Yunnan , Qinghai, Chongqing, Tibet, Sichuan
20 B. coryi Yunnan, China Yunnan
21 B. wilsonae Yunnan, China Yunnan, Sichuan, Tibet, Gansu
22 B. sp. Tibet, China Tibet
23 B. sp. Tibet, China Tibet
Species of barberry identified as alternate hosts for Pst
by artificial inoculation using germinated teliospores in China
16. 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.
Berberis soulieana
B. shensiana
17. Can barberry species be infected by the
stripe rust fungus (Pst) in nature?
It is very common to see pycnia and aecia on
infected Berberis leaves in fields in May.
18. Obtaining of Pst isolates from rust-
infected barberry growing in nature
20 Pst isolates recovered from the infected barberry
4 Pst isolated from 3 barberry species in 2011
16 Pst isolates from 3 barberry species in 2013
collected the infected
Berberis leaves
Single aecium (SA) was
used for inoculation
Recovered SA isolate
on wheat leaf
19. Berberis spp. No. of aecia
No. of isolate Rate (%)
IT0 IT1-2 Pst Pgt Pst Pgt
B. brachypoda 1519 479 1032 2 6 0.13 0.39
B. shensiana 410 35 374 1 0 0.24 0.00
B. soulieana 384 367 16 1 0 0.26 0.00
B. potaninii 742 711 28 0 3 0.00 0.40
B. aggregata 648 633 13 0 2 0.00 0.31
Total 3703 2225 1463 4 11 0.11 0.30
Pst isolates recovered from rust-infected
Berberis spp. in Gansu, and Shaanxi in 2011
*I T0 indicates no any necrosis, or/ and chlorosis on wheat after inoculation,
I T1- 2 indicates necrosis, or/ and chlorosis;
Pst=Puccinia st riif ormis f. sp. trit ici; Pgt =Puccinia graminis f. sp. t rit ici
20. Comparison of infection types on Chinese differential
hosts of eight major races of Pst and the four Pst
isolates recovered from three barberry species in 2011
Virulence tests demonstrated that the infection types of
the four barberry-recovered cultures are different
compared to the major Chinese races.
Cultures
and
races
Origin of cultures
Differential hosts
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
B2011-1 Berberis brachypoda A V A V V V A V A VA V A A V A A A A A
B2011-2 B. brachypoda A V A V V A A V A A V A A A A A A A A
B2011-3 B. shensiana A V A V V A V V A AV V A A AV A A A A A
B2011-4 B. soulieana A V A A V V A V A A A A A V A V A A A
CYA33 Triticum aestivum V V V V V V V V V V V V V V A V A A A
CYA32 T. aestivum V V V V V V V V V V V V V V A V V A A
CYA31 T. aestivum V V V V V V V V V A V V A V A V V A A
CYA30 T. aestivum V V V V V V V V V A V V A A A V V A A
CYA29 T. aestivum V V V V V V V V V A V V A A A V A A A
CYA28 T. aestivum V V V V V V V V V A V A A A A V A A A
CYA23 T. aestivum V V V V A V V V V A V A A A A A A A A
CYA17 T. aestivum VA V A VA A VA V A A A AV A A A A A A A A
Zhao, J., et al. Phytopathology, 2013
21. Berberis spp. No. of aecia
Number of isolate* Rate (%)
IT 0 IT 1-2 Pst Pgt Pst Pgt
B. shensiana 3677 3630 40 4 3 0.11 0.08
B. brachypoda 4477 4406 59 8 4 0.17 0.15
B. sp. (Tibet) 1163 1150 6 4 3 0.34 0.26
Total 9317 9174 105 16 12 0.17 0.13
*I T 0 indicates no any necrosis, or/ and chlorosis on wheat after inoculation,
I T1- 2 indicates necrosis, or/ and chlorosis;
Pst=Puccinia st riif ormis f. sp. trit ici; Pgt =Puccinia graminis f. sp. t rit ici
Pst isolates obtained from the infected Berberis
spp. in Shaanxi, Gansu and Tibet in 2013
22. Berberis
(origin)
Isolate
No.
Chinese differential hosts for Pst
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
20
(ck
)
Race /
race group
U1 A
V
V V V V V V V A V V A V VA A AV A A A V SuG
U2 A V V V V V V V A V
A
V
A A V A A A A A V SuG
B.aggregata
(Gansu)
U3 A V A AV V V V V A V
A
V
A A V A A A A A V SuG
U4 A
V
A V V
A
V
V V V A V V V A V A V A A A V SuG
U5 V V V V V V V V V V V V V V A V V A A V CYR32
U6 A V
A
V
V V V V V A V V V A V A A A A A V SuG
U7 V V V V V V V V A V V
A
V
A V A A A A A V SuG
U8 V V V V V V V V V V V V V V A V V A A V CYR32
B.shensiana
(Shaanxi)
U 9 V V V V V V V V V V V V V V A V V A A V CYR32
U10 V V V V V V V V V V V V V V V V A A A V SuG
U11 A V A V V V V V A
A
V
A
V
A A V A A A A A V SuG
U12 A V A AV V V V V A V
A
V
A A V A A A A A V SuG
U13 A
V
V V V A V V V A V V A A V A A A A A V SuG
3 recovered isolates were identified as race CYR 32.
Other 13 isolates have different virulence patterns as compared
to the major Chinese races , and can be considered as new races.
Chinese differential hosts for Pst
A=avirulent ; V=virulent; AV/VA =segeration of resistant and susceptible; ck=Mingxian 169 susceptible to Pst.
SuG=race group virulence for Suwon 11(YrSu); HyG=race group virulence for Hyrbrid 46 (Yr3b,Yr4b)
CYR= chinese yellow rust
Virulence patterns of 16 recovered isolates on
Chinese differential host sets in 2013
23. Single Aecia
from barberry
Recovered isolate
from single aecium
Single uredium
(Su) isolates
Virulence test on
NILs
Does the sexual reproduction under natural
conditions contribute to virulence variation for
Pst ?
• 120 single-uredium (SU) isolates were make from 16 recovered
isolates from infected barberry.
• Virulence test for each SU isolates on near-isogenic lines.
• Remarkable variation in virulence patterns among the SU isolates.
24. Virulence patterns of single-uredium isolates
from recovered Pst isolate on near-isogenic lines
SU isolate
Yr NILs
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
Yr2 Yr5 Yr6 Yr7 Yr8 Yr9 Yr10 Yr15 Yr17 Yr24 Yr26 Yr27 Yr44 YrSp YrTr1 YrExp2 Yr25 Yr28 CK
U10*
VA A V A A A A A A A AV A V A A A V V V
U10-1 A A V A A V A A A A A A V A A A V V V
U10-2 A A V A A A A A A A A A V A A A V V V
U10-3 A A V A A A A A A A A A V A A A V V V
U10-4 A A V A A V A A A A A A V V A A V V V
U10-5 A A V A A V A A V A A A V V A A V V V
U10-6 V A V AV A A A A V A A A V V A A V V V
U10-7 A A V A A V A A V A A V V V A A V V V
U10-8 A A V A A V A A A A A A V A A A V V V
U10-9 A A V AV A V A A A A A A V A A A V V V
• SU isolates U10-2 and U10-3 share the same virulence patterns,
• Other 7 SU isolates have different virulence patterns.
*U10 isolate recovered from Berberis shensiana in Shaanxi
25. SU
isolate
Yr NILs
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
Yr2 Yr5 Yr6 Yr7 Yr8 Yr9 Yr10 Yr15 Yr17 Yr24 Yr26 Yr27 Yr44 YrSp YrTr1 YrExp2 Yr25 Yr28 CK
U2* A A A AV V A A A A A V A A A A V A A V
U2-1 A A A AV V A A A A A V A A A A V A A V
U2-2 A A A A V A A A AV A V A V A A V A A V
U2-3 A A VA AV V A A A A A V A A A A A A A V
U2-4 A A A A V A A A V A V A V A A V A A V
U2-5 A A AV A V A A A A A V A A A A V A V V
U2-6 A/V A V V V A A A A A V A V A A V A A V
U2-7 A A A A V A A A A A V A A A A V A A V
U2-8 VA A V
A/V
A
V A A A V A V A VA A A V A A V
U2-9 A A AV A V A A A A A V A V A A V A A V
U2-10 A A A A V A A A A A V A A A A V A A V
U2-11 A A AV VA V A A A A A V A A A A V A A V
U2-12 A A AV A V A A A A A V A A A A V A A V
U2-13
A/V
A
A A/V A V A A A V A V A A A A V A A V
U2-14 A/V A A/V
A/V
A
V A A A A A V A A/V A A V V VA V
“/” indicates various infection type on first and second leaf of genotype wheat cultivar. AV, VA= segregation
of resistant and susceptible.
*U2 isolate recovered form Berberis aggregata in Gansu.
• U2-7 and U2-10, share the same virulence patterns,
• Other 12 SU isolates have different virulence patterns.
Virulence patterns of single-uredium isolates
from recovered Pst isolate on near-isogenic lines
26. Single Aecia
from barberry
Recovered isolate
from single aecium
Single uredium
(Su) isolates
Virulence test on
NILs
Does the sexual reproduction under natural
conditions contribute to virulence variation for
Pst?
• The single-uredium isolates originally from aecia on naturally
infected barberry plants consisted of diverse of virulence
patterns, which should be due to sexual reproduction on the
barberry plants.
27. Does infection of stripe rust on Barberry
cause the disease in adjacent wheat?
28. Does infection of stripe rust on Barberry cause
the disease in adjacent wheat?
Mock infection were performed in Gansu, China, by putting
wheat tissues bearing telia on the top of barberry plants in
Middle of April, 2013.
The inoculated barberry plants showed symptom in early May,
2013, and 70 Pst isolates were obtained from mock-inoculated
barberry.
29. Stripe rust appeared on wheat adjacent to the mock-inoculated
barberry plants before the end of May, 2013.
No stripe rust on wheat in the same field far off mock-
inoculated barberry plants, and in other fields at this time.
70 Pst isolates were obtained from nearby wheat.
Does infection of stripe rust on Barberry cause the
disease in adjacent wheat?
30. Comparison of pathotypes on Chinese differential hosts
for Pst isolates from mock-inoculated barberry and
from nearby wheat
Origin Number of
race
Race type
CYR32 CYR31 Guinong 22 Hy46-7 Other racesa
Mock-inoculated
barberry
70 11 1 20 1 37
Nearby wheat 70 8 0 19 8 35
a
Infection type of other races were not matched with those of known Pst races.
The results showed that the races derived from
mock-inoculated barberry matched with those isolated
from nearby wheat.
31. 23 of the 30 tested barberry species show susceptibility to
Pst, indicating that there is a great diversity of potential
alternate hosts for Pst in China.
20 Pst isolates obtained from different barberry species in
the fields indicated that the sexual stage of Pst occurs under
natural conditions.
Virulence tests for single-uredium isolates recovered from a
single aecium demonstrate that the sexual reproduction
contributes to virulence variation for Pst.
Summary
32. Pst must now be considered a macrocyclic fungal pathogen.
33. The earmarked fund for Modern
Agro-industry Technology
Research System in China
National Basic Research
Program of China (973)
Nature Science Foundation of
China
The 111 Project from the
Chinese Ministry of Education
Acknowledgement
Dr. Jie Zhao Prof. Lili Huang
Dr. Hongchang Zhang Dr. Gangming Zhan
Dr. Qingmei Han Dr. Yonghong Zhang
Dr. Dejun Han Dr. Xiaojie Wang
Dr. Jun Guo Dr. Xueling Huang
Dr. Wenming zheng Mrs. Guorong Wei
All PhD and Master students 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