Berry viruses complexes and  what we can do about it       Ioannis E. Tzanetakis          itzaneta@uark.edu             47...
Virus transmission 101
Phylum Arthropoda, Class Insecta, Order Hemiptera
Phylum Arthropoda, Class Insecta
Phylum Arthropoda, Class Arachnida, Order Acariformes
Phylum Nematoda, Class Secernentea, Order Tylenchida                    Nematodes
Kingdom: FungiPhylum: ChytridiomycotaClass: ChytridiomycetesOrder: Incertae sedisFamily: OlpidiaceaeGenus: Olpidium
Kingdom: RhizariaPhylum: CercozoaClass: PlasmodiophoreaOrder: PlasmodiophoridaFamily: PlasmodiophoridaeGenus: Polymyxa
Vectors and plant viruses they transmit
Four modes of virus transmission
Strawberry viruses circa 2003Arabis mosaic – EuropeTomato ringspotRaspberry ringspot – Europe          NematodesStrawberry...
Strawberry decline    A $50M story
The whitefly viruses
BPYV-SPaV double infections
Other strawberry viruses?  Revisited strawberry virus-like diseases.           Chlorotic fleck              LeafrollGoal: ...
Identification, characterization and development        of detection techniques for strawberry viruses*********    Tobacco...
No vector control
Vector control
Blackberry yellow vein disease First observed in 2000 in the Carolinas.Tested for known viruses (RBDV, TRSV etc) – Several...
Tobacco ringspot virus and BYVDBYVD is very similar to what people thought as being TRSVsymptoms TRSV textbook symptoms   ...
New viruses in Rubus in the last 7 yrs16 viruses & virus-like agents were known to infect Rubusbefore we started looking i...
New viruses in Rubus in the last 8 yrs16 viruses & virus-like agents were known to infect Rubusbefore we started looking i...
Same disease-different viruses  AR        MS           NC
Arkansas BYVaV BVY TRSV
Carolinas   BYVaV   BVX   BPYV   INSV   TRSV
Mississippi   BYVaV              TRSV              BVE
How do we tackle BYVD?After identification of all (or almost all) viruses that are involved inthe disease we need to:A. Ma...
What are the viruses present in your area?      The importance of detection• BYVaV - Multistate sample  collection - 35 is...
What are the viruses present in your area?      The importance of detection• BYVaV - Multistate sample  collection - 35 is...
Virus interactions: The BYVaV/BVY storyBVY did not cause symptoms in singleinfections but together with BYVaV theycause BY...
Transmission  Experiment     Trialeurodes abutilonea   Trialeurodes vaporariorum  Experiment 1   4/7                      ...
Alternate hosts                                                                     Number of plantsPlant species         ...
Vector elimination             The BRNV paradigm   New field monitoring     Time of transmissionPermanent tagged plants   ...
Average Aphids/Trap                           0                                2                                      4   ...
Average Aphids/Trap: 2005                      6                                                                          ...
Average Aphids/Trap                 0                                 1                                                   ...
Raspberry crumbly fruit and decline• The Pacific Northwest (PNW) is a primary producer of red  raspberries
Raspberry crumbly fruit and decline• ‘Several cultivars are susceptible to  crumbly fruit disease (drupelets  abortion)• R...
Another virus complex?Important observations suggested that crumbly fruitsymptoms may be increased by additional viruses:1...
RBDV, RLMV and RpLV interactions
RLMV qRT-PCR RLMV titer in single and mixed infections over time                                        ThresholdΔRn      ...
RpLV qRT-PCR RpLV titer in single and mixed infections over time         int. control                                     ...
RBDV titer enhanced in co-infections with RLMV
RBDV titer enhanced in co-infections with RLMV  RBDV titer increase verified by conventional methods               ELISA  ...
Mixed virus infections affect on plant growth           and fruit crumbliness •   H Control                 Raspberry viru...
Plant Growth       Establishment (2010)Oct.                                                  Height                     14...
Plant Growth (2011)                                           July                                  Height                ...
Plant Growth (2011)                                                                                              October  ...
Crumbly Fruit                    No. Drupelets                                             Weight120                      ...
Crumbly fruit                               Firmness                                                                      ...
Virus Incidence in ‘Meeker’ Fields   Northern Washington             Southern  Field Age   RLMV   RpLV      Washington/Ore...
Crumbly Fruit ScoutingCrumbly fruit and virus incidence in Washington        Field   Crumbliness          Virus incidence ...
Insects in Traps (2011)Empoasca fabae was sporadic                                            Raspberry aphid A. agathonic...
RLMV spread in the fieldFour fields being monitored for virus spreadField 4                                     26 %    (2...
Control Strategies1. Think long term, identify potential risks of a site2. Start with clean plants3. Identify and diagnose...
The importance of clean plants• Better establishment
The importance of clean plants• Better establishment• Longer life of plantings
The importance of clean plants• Better establishment• Longer life of plantings• Fewer disease problems/Reduce risk of intr...
National Clean Plant NetworkA federally-coordinated effort to secure high quality virus-tested plants for clonally propaga...
NCPN Supported Clean Plant Centers                   WSU                       USDA at OSU                              ...
Berry Clean Plant Centers USDA at OSU    UC-Davis                           NCSU                              UAF
Why care???
The story:Propagation from an existing plot10 ton/acre =$30,000/yearLatent infections with Blueberry scorchThe result?Remo...
Florida 2013
Start clean!!!!
The teamThe berry virus consortium:16 individuals from UA, NCSU, USDA-ARS, MSU, UGABindu Poudel, Diego Quito, Danielle Lig...
Berry viruses complexes and what we can do about it
Berry viruses complexes and what we can do about it
Berry viruses complexes and what we can do about it
Berry viruses complexes and what we can do about it
Berry viruses complexes and what we can do about it
Berry viruses complexes and what we can do about it
Berry viruses complexes and what we can do about it
Berry viruses complexes and what we can do about it
Berry viruses complexes and what we can do about it
Berry viruses complexes and what we can do about it
Upcoming SlideShare
Loading in …5
×

Berry viruses complexes and what we can do about it

1,226 views

Published on

Ioannis E. Tzanetakis
itzaneta@uark.edu 479-575-3180

Published in: Education
0 Comments
0 Likes
Statistics
Notes
  • Be the first to comment

  • Be the first to like this

No Downloads
Views
Total views
1,226
On SlideShare
0
From Embeds
0
Number of Embeds
2
Actions
Shares
0
Downloads
35
Comments
0
Likes
0
Embeds 0
No embeds

No notes for slide

Berry viruses complexes and what we can do about it

  1. 1. Berry viruses complexes and what we can do about it Ioannis E. Tzanetakis itzaneta@uark.edu 479-575-3180
  2. 2. Virus transmission 101
  3. 3. Phylum Arthropoda, Class Insecta, Order Hemiptera
  4. 4. Phylum Arthropoda, Class Insecta
  5. 5. Phylum Arthropoda, Class Arachnida, Order Acariformes
  6. 6. Phylum Nematoda, Class Secernentea, Order Tylenchida Nematodes
  7. 7. Kingdom: FungiPhylum: ChytridiomycotaClass: ChytridiomycetesOrder: Incertae sedisFamily: OlpidiaceaeGenus: Olpidium
  8. 8. Kingdom: RhizariaPhylum: CercozoaClass: PlasmodiophoreaOrder: PlasmodiophoridaFamily: PlasmodiophoridaeGenus: Polymyxa
  9. 9. Vectors and plant viruses they transmit
  10. 10. Four modes of virus transmission
  11. 11. Strawberry viruses circa 2003Arabis mosaic – EuropeTomato ringspotRaspberry ringspot – Europe NematodesStrawberry latent ringspot-EuropeTomato black ring –EuropeStrawberry crinkleStrawberry mild yellow edge AphidsStrawberry mottleStrawberry vein bandingFragaria chiloensis latent – Chile PollenTobacco streak*Strawberry pallidosis Whiteflies*Beet pseudo-yellows
  12. 12. Strawberry decline A $50M story
  13. 13. The whitefly viruses
  14. 14. BPYV-SPaV double infections
  15. 15. Other strawberry viruses? Revisited strawberry virus-like diseases. Chlorotic fleck LeafrollGoal: Identify unknown viruses that may contribute to the decline.
  16. 16. Identification, characterization and development of detection techniques for strawberry viruses********* Tobacco streak virus* Strawberry crinivirus 3* Strawberry crinivirus 4
  17. 17. No vector control
  18. 18. Vector control
  19. 19. Blackberry yellow vein disease First observed in 2000 in the Carolinas.Tested for known viruses (RBDV, TRSV etc) – Several viruseswere found but none consistently associated with symptoms.
  20. 20. Tobacco ringspot virus and BYVDBYVD is very similar to what people thought as being TRSVsymptoms TRSV textbook symptoms Single TRSV-infectionAre symptoms cv. dependent? The majority of plantsinfected with TRSV are symptomless
  21. 21. New viruses in Rubus in the last 7 yrs16 viruses & virus-like agents were known to infect Rubusbefore we started looking into Rubus complexes – We nowhave over 40…New Rubus virusesBlackberry yellow vein associated virusBlackberry virus EBlackberry virus XBlackberry virus YBlackberry virus ZBeet pseudo yellows virusBlackberry yellow mottle virusBlackberry chlorotic ringspot virusStrawberry necrotic shock virusBlack raspberry necrosis virusRaspberry leaf mottle virusRubus canadensis virus -1Impatiens necrotic spot virusRaspberry latent virusetc…..
  22. 22. New viruses in Rubus in the last 8 yrs16 viruses & virus-like agents were known to infect Rubusbefore we started looking into Rubus complexes – We nowhave over 40…New Rubus virusesBlackberry yellow vein associated virusBlackberry virus E Tests are available for all the new virusesBlackberry virus XBlackberry virus YBlackberry virus ZBeet pseudo yellows virusBlackberry yellow mottle virusBlackberry chlorotic ringspot virusStrawberry necrotic shock virusBlack raspberry necrosis virusRaspberry leaf mottle virusRubus canadensis virus -1Impatiens necrotic spot virusRaspberry latent virusetc…..
  23. 23. Same disease-different viruses AR MS NC
  24. 24. Arkansas BYVaV BVY TRSV
  25. 25. Carolinas BYVaV BVX BPYV INSV TRSV
  26. 26. Mississippi BYVaV TRSV BVE
  27. 27. How do we tackle BYVD?After identification of all (or almost all) viruses that are involved inthe disease we need to:A. Make sure that mother plants are being tested for the new viruses before they are propagated.B. Identify virus combinations that can cause BYVD.C. Identify virus vectors.D. Find alternative hosts of the viruses in the field.E. Minimize or eliminate BYVD by eliminating the weakest link, the virus vector(s) that is the easiest to control.
  28. 28. What are the viruses present in your area? The importance of detection• BYVaV - Multistate sample collection - 35 isolates• i
  29. 29. What are the viruses present in your area? The importance of detection• BYVaV - Multistate sample collection - 35 isolates• i 1 2 3 4 5 6 7 8 9• Detection 100% identity
  30. 30. Virus interactions: The BYVaV/BVY storyBVY did not cause symptoms in singleinfections but together with BYVaV theycause BYVD.In mixed infections, BVY knocks downconcentration of BYVaV to about 0.1%compared to titer in single infections.In mixed infections, they can cause death of fruiting canes. Susaimuthu et al.,2008. Plant Disease 92:1288-1292
  31. 31. Transmission Experiment Trialeurodes abutilonea Trialeurodes vaporariorum Experiment 1 4/7 3/9 Experiment 2 5/8 1/8 Experiment 3 3/10 3/10 Total 12/25 7/27• Both whitefly species transmitted the virus at a rate >30%
  32. 32. Alternate hosts Number of plantsPlant species Scientific name Family testedGarden vetch Vicia sativa Fabaceae 16Virginia creeper Parthenocissus quinquefolia Vitaceae 16Red clover Trifolium pretense Fabaceae 16Wild garlic Allium vineale Amaryllidaceae 16Creeping woodsorrel Oxalis corniculata Oxalidaceae 16Carolina geranium Geranium carolinianum Geraniaceae 16Curly dock Rumex crispus Polygonaceae 16Dandelion Taraxacum officinale Asteraceae 16Tall fescue Festuca arundinacea Poaceae 16Wild wheat Avena fatua Poaceae 16Grapes Vitis vinifera Vitaceae 16Peach Prunus persica Rosaceae 16Blueberry Vaccinium spp. Ericaceae 16Shepherd’s purse Capsella bursa-pastoris Brassicaceae 16Nutsedge Cyperus spp. Cyperaceae 16Horsenettle Solanum carolinense Solanaceae 16Common ragweed Ambrosia artemisiifolia Asteraceae 16Tree of heaven Ailanthus altissima Simaroubaceae 16Apple Malus spp. Rosaceae 200Rose Rosa multiflora Rosaceae 40Carpetweed Mollugo verticillata Molluginaceae 16Amaranthus Amaranthus spp. Amaranthaceae 16Poor joe Diodia teres Rubiaceae 16Ground cherry Physalis spp. Solanaceae 16Sorghum Sorghum spp. Poaceae 16
  33. 33. Vector elimination The BRNV paradigm New field monitoring Time of transmissionPermanent tagged plants High incidence of virus Potted plants Rotated every month P L H D O K G C N J F B M I E A
  34. 34. Average Aphids/Trap 0 2 4 6 8 10 12 14 4-May 11-May 18-May 25-MayField 1 1-Jun 8-Jun 15-Jun 22-Jun Date 29-JunField 2 6-Jul 13-Jul 20-Jul Average Aphids/Trap: 2004 27-Jul 3-Aug 10-Aug 17-Aug 40 50 60 70 80 90 100 110 o Max Daily Temp ( F)Field 3 Field 3 Field 2 Field 1 M ax D aily T emp
  35. 35. Average Aphids/Trap: 2005 6 100 5 90Average Aphids/Trap Max Daily Temp ( F) o 4 80 Field 1 Field 2 3 70 Field 3 Maximum Daily Temp 2 60 1 50 0 40 5-Jul 19-Jul 1-Mar 15-Mar 29-Mar 12-Apr 26-Apr 7-Jun 21-Jun 2-Aug 16-Aug 30-Aug 13-Sep 27-Sep 10-May 24-May 11-Oct 25-Oct Date Field 1 Field 2 Field 3 Nearly 100% transmission in three years!
  36. 36. Average Aphids/Trap 0 1 2 3 4 5 6 7 10-May 17-May 24-May 31-May 2-Jun 7-Jun 14-Jun 21-Jun 17-Jun 28-Jun 5-Jul 12-Jul 6-Jul 19-Jul 20-Jul 26-Jul 2-Aug 9-Aug 16-AugDate 3-Aug 18-Aug 23-Aug 30-Aug 2-Sep 6-Sep 13-Sep 20-Sep 16-Sep 27-Sep Average Aphids/Trap vs. Positives: 2004 4-Oct 30-Sep 11-Oct 18-Oct 25-Oct 1-Nov 14-Oct 28-Oct 0 2 4 6 8 10 12 14 16 Time of Transmission # of BRDaV Positives # of positives Average Aphids: 2004
  37. 37. Raspberry crumbly fruit and decline• The Pacific Northwest (PNW) is a primary producer of red raspberries
  38. 38. Raspberry crumbly fruit and decline• ‘Several cultivars are susceptible to crumbly fruit disease (drupelets abortion)• Raspberry bushy dwarf virus (RBDV), a pollen-borne idaeovirus was considered the causal agent of crumbly fruit• Still, in many cases RBDV single infections did not cause symptoms
  39. 39. Another virus complex?Important observations suggested that crumbly fruitsymptoms may be increased by additional viruses:1. The disorder is more severe in cool areas with high populations of the large raspberry aphid Amphorophora agathonica2. Two additional viruses found in severely affected fields, Raspberry leaf mottle (RLMV) and Raspberry latent (RpLV)
  40. 40. RBDV, RLMV and RpLV interactions
  41. 41. RLMV qRT-PCR RLMV titer in single and mixed infections over time ThresholdΔRn Amp. Cycle
  42. 42. RpLV qRT-PCR RpLV titer in single and mixed infections over time int. control ThresholdΔRn Amp. Cycle
  43. 43. RBDV titer enhanced in co-infections with RLMV
  44. 44. RBDV titer enhanced in co-infections with RLMV RBDV titer increase verified by conventional methods ELISA RT-PCR (20 cycles)
  45. 45. Mixed virus infections affect on plant growth and fruit crumbliness • H Control Raspberry virus .... what...? • D RBDV - Dwarf • M RLMV - Mottle • L RpLV - Latent • DM RBDV + RLMV • DL RBDV + RpLV • ML RLMV + RpLV • DML RBDV + RLMV + RpLV
  46. 46. Plant Growth Establishment (2010)Oct. Height 140.00 120.00 109.03 100.00 80.00 65.43 63.83 60.00 44.90 48.93 cm 33.49 32.00 40.00 26.09 20.00 a cd b b d bc cd cd 0.00 H D M L DM DL LM DML TreatmentsOct. Cane diameter 12.00 10.07 10.00 8.18 8.00 7.02 6.30 5.94 5.74 5.73 mm 6.00 5.41 4.00 2.00 a bc d cd d cd d b 0.00 H D M L DM DL LM DML Treatments
  47. 47. Plant Growth (2011) July Height Cane diameter 70.00 9.00 8.06 7.81 7.72 55.57 8.00 7.34 7.18 7.34 6.99 60.00 51.27 51.03 50.73 48.50 7.00 6.05 50.00 44.60 42.13 6.00 mm 40.00cm 5.00 30.20 30.00 4.00 3.00 20.00 2.00 10.00 a b b b d b c c 1.00 a a a b c b b b 0.00 0.00 H D M L DM DL LM DML H D M L DM DL LM DML Treatments Treatments
  48. 48. Plant Growth (2011) October Height Cane diameter 3 14 12.79 2.60 11.79 11.52 11.83 2.25 2.25 12 10.77 10.82 10.21 10.03 2.5 2.17 2.12 1.97 1.93 1.89 10 2 8m 1.5 mm 6 1 4 0.5 a b b b cd bc cd d 2 a b b b c bc c c 0 0 H D M L DM DL ML DML H D M L DM DL ML DML Treatments Treatments
  49. 49. Crumbly Fruit No. Drupelets Weight120 4.000 94 95 3.35100 89 75 76 3.000 2.74 2.87 2.5780 2.43 2.33 64 59 66 2.08 2.0560 g 2.00040 1.00020 a c a a b c b c a c b b bc cd cd d 0 0.000 H D M L DM DL LM DML H D M L DM DL LM DML Treatments Treatments
  50. 50. Crumbly fruit Firmness D 90.000 80.000 73.54 65.58 70.000 58.54 61.38 61.83 61.46 60.000 46.38 49.15 50.000g 40.000 30.000 20.000 10.000 a c bc b bc d bc d 0.000 H D M L DM DL LM DML Treatments H DML DML
  51. 51. Virus Incidence in ‘Meeker’ Fields Northern Washington Southern Field Age RLMV RpLV Washington/Oregon (years) (%) (%) Field Age RLMV RpLV 1 4 0 (years) (%) (%) 1 30 0 1 0 0 1 10 0 5 40 20 2 58 21 2 0 0 6 0 20 2 0 0 7 8 17 2 6 0 8 19 0 2 16 0 8 27 0 3 31 6 3 6 0 3 13 0 3 50 0 4 19 6 4 13 0 5 69 0 5 90 80 5 100 75 5 44 6 5 100 17 6 70 25 6 100 6 6 100 12 7 100 6 8 100 46
  52. 52. Crumbly Fruit ScoutingCrumbly fruit and virus incidence in Washington Field Crumbliness Virus incidence Age 0: normal % 3: severe 0 1 2 3 RBDV RLMV RpLV 4 1 44 25 0 4 2 100 100 0 4 3 93 100 7 5 3 100 100 40 6 3 92 96 40Crumbliness Crumbliness Crumblinessrate 1 rate 2 rate 3
  53. 53. Insects in Traps (2011)Empoasca fabae was sporadic Raspberry aphid A. agathonica predominant insect Average # aphids / 10 8Few numbers of 7 Harvest clean-upMacropsis fuscula 6 sprays applied 5 leaves 4 3 2 1 0 1-Apr 16-May 30-Jun 14-Aug 28-Sep Lightle, unpublished data
  54. 54. RLMV spread in the fieldFour fields being monitored for virus spreadField 4 26 % (24 %) 50 % 4% (40 %) 44 %Field 3 21 % (12 %) 33 %Field 2 16 % (16 %) 32 %Field 1 year 0 year 1 year 2 year 3 Age of Field
  55. 55. Control Strategies1. Think long term, identify potential risks of a site2. Start with clean plants3. Identify and diagnose problems early4. Implement control strategies ASAP5. If a virus complex is involved - identify viruses present and which are the easiest to control
  56. 56. The importance of clean plants• Better establishment
  57. 57. The importance of clean plants• Better establishment• Longer life of plantings
  58. 58. The importance of clean plants• Better establishment• Longer life of plantings• Fewer disease problems/Reduce risk of introducing new viruses to a region or field
  59. 59. National Clean Plant NetworkA federally-coordinated effort to secure high quality virus-tested plants for clonally propagated crops.NCPN MissionThe NCPN provides high quality asexually propagatedplant material free of targeted plant pathogens andpests that cause economic loss to protect theenvironment and ensure the global competitiveness ofspecialty crop producers in the United States.
  60. 60. NCPN Supported Clean Plant Centers WSU  USDA at OSU Cornell   UC-Davis MO State U.   NCSU  UCR    UAZ UAF Clemson  Auburn LSU  FAMU    UHM FL-Dept. Ag.  TAMU
  61. 61. Berry Clean Plant Centers USDA at OSU UC-Davis  NCSU  UAF
  62. 62. Why care???
  63. 63. The story:Propagation from an existing plot10 ton/acre =$30,000/yearLatent infections with Blueberry scorchThe result?Removal of infected materialCumulative loss: ~ 100,000/acre
  64. 64. Florida 2013
  65. 65. Start clean!!!!
  66. 66. The teamThe berry virus consortium:16 individuals from UA, NCSU, USDA-ARS, MSU, UGABindu Poudel, Diego Quito, Danielle Lightle, Anne Halgren,James Susaimuthu

×