![Page 1: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/1.jpg)
Orchid virus diseases in Taiwan and their control strategies
![Page 2: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/2.jpg)
Characteristics of virus diseases
Causing systemic infection
Not curable once infected
Reduce growth vigor and quality
![Page 3: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/3.jpg)
Viruses known to infect orchids in the literature
In total, more than 30 viruses recorded in literature.
17 with conclusive taxonomic status.
![Page 4: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/4.jpg)
Rod-shapedTobamovirus
Odontoglossum ringspot virus (ORSV)
Tobacco mosaic virus-orchid strain (TMV-orchid)
Potexvirus
Cymbidium mosaic virus (CymMV)
Tobravirus
Tobacco rattle virus (TRV)
Potyvirus
Bean yellow mosaic virus (BYMV)
Dendrobium mosaic virus (DenMV)
Clover yellow vein virus (CYVV)
Turnip mosaic virus (TuMV)
Vanilla mosaic virus (VaMV)
Phalaenopsis chlorotic spot virus (PhCSV)
Closterovirus
Dendrobium vein necrosis virus (DVMV)
![Page 5: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/5.jpg)
IsometricTombusvirus
Cymbidium ringspot virus (CyRSV)
Carmovirus
Carnation mottle virus (CarMV)
Cucumovirus
Cucumber mosaic virus (CMV)
Nepovirus
Tomato ringspot virus (TomRSV)
Tospovirus
Tomato spotted wilt virus (TSWV)
Impatiens necrotic spot virus (INSV)
Capsicum chlorosis virus (CaCV)
Unassigned
Orchid fleck virus (OFV)
![Page 6: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/6.jpg)
1. Odontoglossum ringspot virus (ORSV)
2. Cymbidium mosaic virus (CymMV)
3. Cucumber mosaic virus (CMV)
4. Capsicum chlorosis virus-phalaenopsis isolate (CaCV)
5. Phalaenopsis chlorotic spot virus (PhCSV)
6. Bean yellow mosaic virus (BYMV)
7. Turnip mosaic virus (TuMV)
8. Carnation mottle virus (CarMV)
9. Orchid fleck virus (OFV)
Orchid viruses found in Taiwan
![Page 7: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/7.jpg)
Odontoglossum ringspot virus (ORSV)
Tobamovirus, rigid rods, 300 nm in length
Heat stable, tolerant up to 95℃
Highly stable in vitro, last over years (10 yrs)
Transmitted by mechanical contact, no vector known
![Page 8: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/8.jpg)
Cymbidium mosaic virus (CymMV)
Potexviruses, filamentous particles (450 nm in length)
Transmitted by contact, no vector known.
Not as stable in vitro as ORSV,
but very stable compared to other
plant viruses.
![Page 9: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/9.jpg)
• Symptoms: yellow striping on leaves.
• Cucumovirus
• Isometric particles
• Mechanically transmissible and also
by aphids with non-persistent manner.
• Occurs more frequently on orchids
grown in open fields
Cucumber mosaic virus (CMV)
![Page 10: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/10.jpg)
• Symptoms: chlorotic ringspot on leaves.
• Tospovirus
• Isometric enveloped particles.
• Serologically related to serogroup IV
tospoviruses.
• Mechanically transmissible and also
by thrips with persistent manner.
• Species of vector thrips is not known yet.
Capsicum chlorosis virus (CaCV)
Incorrectly called “Taiwan virus”
![Page 11: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/11.jpg)
![Page 12: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/12.jpg)
Phalaenopsis chlorotic spot virus (PhCSV)
or Basella rugose mosaic virus (BaRMV) ?Potyvirus
Mechanical transmissible and also
by aphid with non-persistent manner
Induce chlorotic spot symptoms on
leaves
Need to confirm its taxonomic status.
![Page 13: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/13.jpg)
Turnip mosaic virus (TuMV)
Potyvirus
Transmitted mainly by aphids with
non-persistent manner.
Natural hosts for TuMV are cruciferous
vegetables.
Found only once in Taiwan.
![Page 14: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/14.jpg)
Carnation mottle virus (CarMV)
Carmovirus, Tombusviridae
Very stable in vitro and remains infective
for long period of time.
Can easily contaminate working benches
and laminar flow.
Possibly induce chlorotic spots on Phalaenopsis.
![Page 15: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/15.jpg)
Orchid fleck virus (OFV)
Rhabdovirus The unassigned viruses
Bullet shaped virus particles
Transmitted by mites (Brevipalpus californicus)
First reported in Japan.
Occurring also in Brazil,
Germany, USA and Australia
![Page 16: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/16.jpg)
OFV has been eradicated since 2005
![Page 17: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/17.jpg)
Table. Result of surveillance of Orchid fleck virus in Taiwan
since 2005 to 2007.
Result
Source Species No. of sample Detection method Date RT-PCR ELISA
Lu-ku cymbidium 37 ELISA 94.12.08 - 0/37
Yu-chi cymbidium >5000 V. I. 95.10.07 0 0
Chai-yi phalaenopsis 2 RT-PCR 95.10.23 0/2 -
TDAIS cymbidium 3 RT-PCR, ELISA 96.01.26 0/3 0/3
TDAIS cymbidium 16 RT-PCR, ELISA 96.01.26 0/16 0/16
Yu-chi cymbidium >5000 V. I. 96.01.26 0 0
Pin-tung phalaenopsis 1 RT-PCR 96.04.10 0/1 -
Hou-Li oncidium 5 RT-PCR 96.08.22 0/5 -
Pu-Li oncidium 1 RT-PCR 96.09.28 0/1 -
V.I. : Visual inspection
![Page 18: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/18.jpg)
Symptoms induced by ORSV on Phalaenopsis spp.
![Page 19: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/19.jpg)
ORSV symptoms on oncidiums
![Page 20: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/20.jpg)
ORSV symptoms on other orchids
![Page 21: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/21.jpg)
CymMV symptoms on Phalaenopsis spp.
![Page 22: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/22.jpg)
CymMV symptoms on Oncidiums
![Page 23: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/23.jpg)
CymMV symptoms on other orchids
![Page 24: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/24.jpg)
Symptoms on Phalaenopsis induced
by dual infection of ORSV and CymMV
![Page 25: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/25.jpg)
ORSVCymMV
![Page 26: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/26.jpg)
![Page 27: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/27.jpg)
Symptom expression:
1. Type of infected virus?
2. Single or mixed infection?
3. Natural resistance of cultivar?
4. Age of plant?
5. Environment factor?
6. Stress?
![Page 28: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/28.jpg)
1. Transmission from infected mother plants by vegetative
propagation including tissue culture.
Mode of transmission and virus sources
![Page 29: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/29.jpg)
![Page 30: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/30.jpg)
2. Seed-borne transmission
Mode of transmission and virus sources
![Page 31: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/31.jpg)
Survey of virus infection in
Phalaenopsis seedlings
Sources No. of Virus infection rate
of flask flask ORSV CyMV O+C (%)
A-W 9 1 0 0 0.7
B-6106 10 3 3 1 4.7
C-00110 10 7 6 3 10.7
D-34010 10 11 58 7 50.7
Virus infection rate of Phalaenopsis seedlings
propagated from tree seeds
![Page 32: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/32.jpg)
Tissue
fibers
Seeds Virus concentration in different parts of
Phalaenopsis fruit capsule assayed by ELISA
A B C D
ORSV CyMV ORSV CyMV ORSV CyMV ORSV CyMV
Capsuls 0.065 * * * 0.315 1.108 0.589 0.060
Cotton fiber with seeds 0.112 * * * 0.071 1.701 * 0.043
Cotton fiber 0.148 * * * 0.086 0.045 * 0.033
Seed wash 1 time with
ddH2O
0.077 1.574 0.287 2.315 0.108 0.049 0.335 0.085
Seed wash 2 times with
ddH2O
0.049 2.545 0.260 0.592 0.142 0.054 0.569 0.058
Seed wash 3 times with
ddH2O
0.024 1.791 0.421 0.521 0.098 0.058 0.178 0.072
Seed wash 4 times with
ddH2O
0.063 2.955 0.516 0.276 0.096 0.102 0.047 0.076
Seed wash 5 times with
ddH2O
0.091 1.598 1.103 0.496 0.025 0.101 0.080 0.060
![Page 33: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/33.jpg)
Tissue
fibers
Seeds
1. ORSV and CyMV are only contaminating on the surface of seeds
instead of infecting the seeds.
2. Virus infected mother plants will produce infected capsules with
high concentration of viruses on the seeds and the fibers.
3. Seeding the infected fibers together with the seeds will eventually
result infected seedlings.
4. Infection rate of the seedlings is variable depending
of the workers during subculture of the seedling.
Mechanisms for seed transmission of Phalaenopsis orchids
![Page 34: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/34.jpg)
3. Virus contamination during tissue culture passages.
Mode of transmission and virus sources
![Page 35: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/35.jpg)
4. Virus contamination during transplanting of plantlets
Mode of transmission and virus sources
![Page 36: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/36.jpg)
5. Transmission by mechanical
contact during culturation.
Mode of transmission and virus sources
![Page 37: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/37.jpg)
![Page 38: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/38.jpg)
7. Transmission by insect or other pests.
Mode of transmission and virus sources
![Page 39: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/39.jpg)
How to produce virus clean orchid plantlets
1. Screening for non-infected plants before
mericloning propagation
a. Collect true to type and symptomless plants.
b. Maintain in a isolated house and keep them well separated.
c. Apply virus indexing tests such as ELISA or RT-PCR on the plants.
d. Eliminate any detected plants from the isolated house.
e. Perform virus check again after at least one month later.
f. Conduct mericloning on the doubly checked plants.
g. Keep the doubly checked plants as mother stocks and apply virus
check at least once every year.
![Page 40: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/40.jpg)
Special concerns:
1. Apply sensitive, dependable and reproducible
virus indexing techniques.
2. Use the right sampling method.
3. Repeats and alternatives.
ELISA, RT-PCR, Biochips, Immuno-strips (Rapid test), etc.
![Page 41: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/41.jpg)
(Enzyme linked immuno-sorbent assay, ELISA)
Naktuinbouw
1. Sensitive
2. Specific
3. Efficient
4. Reproducibe
5. Objective
6. Cost effective
• The most widely used plant virus indexing technique since 1977.
![Page 42: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/42.jpg)
-0.4-0.20.00.20.40.60.81.01.21.41.61.82.02.22.42.62.83.03.23.43.63.84.0
93.08 93.09 93.10 93.11 93.12 94.01 94.02 94.03 94.04 94.05 94.06 94.07 94.08 94.09
-0.4-0.20.00.20.40.60.81.01.21.41.61.82.02.22.42.62.83.03.23.43.63.84.0
93.08 93.09 93.10 93.11 93.12 94.01 94.02 94.03 94.04 94.05 94.06 94.07 94.08 94.09
0.00.20.40.60.81.01.21.41.61.82.02.22.42.62.83.03.23.43.63.84.04.2
93.08 93.09 93.10 93.11 93.12 94.01 94.02 94.03 94.04 94.05 94.06 94.07 94.08 94.09
-0.2
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
1.8
2.0
2.2
2.4
2.6
2.8
93.08 93.09 93.10 93.11 93.12 94.01 94.02 94.03 94.04 94.05 94.06 94.07 94.08 94.09
0.00.20.40.60.81.01.21.41.61.82.02.22.42.62.83.03.23.43.63.84.04.2
93.08 93.09 93.10 93.11 93.12 94.01 94.02 94.03 94.04 94.05 94.06 94.07 94.08 94.09
-0.20.00.20.40.60.81.01.21.41.61.82.02.22.42.62.83.03.23.43.63.84.04.2
93.08 93.09 93.10 93.11 93.12 94.01 94.02 94.03 94.04 94.05 94.06 94.07 94.08 94.09
-0.20.00.20.40.60.81.01.21.41.61.82.02.22.42.62.83.03.23.43.63.84.04.2
93.08 93.09 93.10 93.11 93.12 94.01 94.02 94.03 94.04 94.05 94.06 94.07 94.08 94.09
-0.20.00.20.40.60.81.01.21.41.61.82.02.22.42.62.83.03.23.43.63.84.04.2
93.08 93.09 93.10 93.11 93.12 94.01 94.02 94.03 94.04 94.05 94.06 94.07 94.08 94.09
-0.4-0.20.00.20.40.60.81.01.21.41.61.82.02.22.42.62.83.03.23.43.63.84.04.2
93.08 93.09 93.10 93.11 93.12 94.01 94.02 94.03 94.04 94.05 94.06 94.07 94.08 94.09
Fluctuation of virus concentration in orchid tissue as
determined by ELISA within one year.
![Page 43: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/43.jpg)
ELISA O+C+ O- O+C+ O- C+ HO+ C+
400 bp ORSV
300 bp CyMV
One-tube multiplex RT-PCR assays
Tissue cultureField plant
Detect CymMV and ORSV simultaneously
![Page 44: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/44.jpg)
M CaCV Cy Or C+O COA H
CymMV
ORSV
CaCV
ATPas
One-step multiplex RT-PCR for simultaneous detection of
ORSV, CymMV and CaCV
CaCV
Taiwan virus
![Page 45: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/45.jpg)
Probe
B
B
B
AP
Streptavidin-AP
AP
Biotin labeled PCR product
A
N N
N N
O O
C
C
Biochip detection technology
![Page 46: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/46.jpg)
ELISA CyMV
ORSV
0.165
0.099
0.109
0.069
0.005
0.024
0.219
1.951
H CK B C D. CK
Biochips
RT-PCR
A
N N
N N
O O
C
C
CyMV
ORSV
ATPase
Biochip system for simultaneous detection of
ORSV and CymMV.
![Page 47: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/47.jpg)
Hb CKCymMV
CaCV
ATPaeORSV
設計圖
M CaCV Cy Or C+O COA H
CaCV CymMV ORSV C+O C+O+A H
Biochip system for simultaneous detection of
ORSV, CymMV and CaCV
CaCV
![Page 48: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/48.jpg)
1. Apply sensitive, dependable and reproducible
virus indexing techniques.
2. Use the right sampling method.
3. Repeats and alternatives.
Special concerns:
Leaves, roots, or spikes.
and How to sample.
Do not depend on one single test.
Use different techniques to confirm.
![Page 49: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/49.jpg)
Timing and techniques for Phalaenopsis virus indexing
in Taiwan.
Mother
stocks
Young
plantletsMother
flasks
Tissue
cultured
plantlets
ELISA PCR PCR PCR / ELISA ELISA
![Page 50: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/50.jpg)
2. Take special precaution to prevent clean mother stocks
from re-infection by viruses.
Maintaining clean mother stocks.
Executing sanitation procedures.
Cleaning of growing environment, tools, clothes and
worker’s hands appropriately
Special concerns:
![Page 51: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/51.jpg)
Maintaining in isolated clean room
![Page 52: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/52.jpg)
![Page 53: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/53.jpg)
1. Producing virus clean orchids is a win-win situation for
propagators and finished plants providers.
2. All partners in the orchid production chain should implement
serious sanitation and strict production procedures to prevent
virus spread.
3. However, it is difficult for nurseries to guarantee absolutely
clean flowering orchids considering the high possibilities of re-
infection during lengthy growing period.
4. Orchid consumers or down stream growers should compromise
and accept the reality that certain percentage of virus infection in
the finished flowering plants would not affect their mutual
benefits.
Concluding remarks:
![Page 54: Orchid virus diseases in Taiwan and their control strategies · How to produce virus clean orchid plantlets 1. Screening for non-infected plants before mericloning propagation a](https://reader034.vdocuments.us/reader034/viewer/2022042316/5f0585c77e708231d41361a0/html5/thumbnails/54.jpg)
Concluding remarks:
5. This understanding and compromise will certainly help the harmony
of international orchid trading.
6. It should be emphasized again that, in the long run, screening of
virus-free stocks before mass propagation is a must for all orchid
propagators.
7. No companies can overlook this common demand and still maintain
their competitiveness in the orchid industry around the world.
THANK YOU