slide1 n.
Download
Skip this Video
Loading SlideShow in 5 Seconds..
Molecular characterization of plant viruses infecting potato and vegetables in Iraq PowerPoint Presentation
Download Presentation
Molecular characterization of plant viruses infecting potato and vegetables in Iraq

Loading in 2 Seconds...

play fullscreen
1 / 19

Molecular characterization of plant viruses infecting potato and vegetables in Iraq - PowerPoint PPT Presentation


  • 167 Views
  • Uploaded on

Molecular characterization of plant viruses infecting potato and vegetables in Iraq. NAWRES AL_KUWAITY, SUSAN SEAL and M. N. MARUTHI Natural Resources Institute, University of Greenwich at Medway, Chatham, Maritime, Kent, ME4 4TB, UK. Introduction:.

loader
I am the owner, or an agent authorized to act on behalf of the owner, of the copyrighted work described.
capcha
Download Presentation

PowerPoint Slideshow about 'Molecular characterization of plant viruses infecting potato and vegetables in Iraq' - jenny


An Image/Link below is provided (as is) to download presentation

Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author.While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.


- - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - -
Presentation Transcript
slide1

Molecular characterization of plant viruses infecting potato and vegetables in Iraq

NAWRES AL_KUWAITY, SUSAN SEAL and M. N. MARUTHINatural Resources Institute, University of Greenwich at Medway, Chatham, Maritime, Kent, ME4 4TB, UK

introduction
Introduction:
  • Potato and vegetables are grown in Iraq and have significant economic importance as they are used for daily consumption and a source of income.
  • Plant viruses are one of the factors that limit the production of potato and vegetables worldwide.
  • A number of viruses have been studied in Iraq using conventional methods. However there is no molecular information about plant viruses from Iraq.
slide4

Objectives:

  • To identify plant viruses occurring in potato and vegetable samples collected from Iraqi fields by PCR/RT-PCR using genus/family specific degenerate primers.
  • To study the relatedness of viruses isolated from Iraq to other viruses from different geographical regions by phylogenetic analyses.
slide5

The following virus groups/families were selected to screen for, in potato and vegetables based on their economic importance:

  • 1. Potyviridae
  • 2. Geminiviridae
  • 3. Carlaviruses
  • 4. Potexviruses
  • 5. Alfamoviruses
  • 6. Cucumoviruses
  • 7. Tobamoviruses
  • 8. Tombusviruses

About 60% of known plant viruses are members of these families

Previously reported in Iraq

Symptoms on certain samples

slide7

Gel recovery and extraction

RT-PCR using genus/family specific primer sets

Dried samples

Cloning into pGEMT-vector

DNA fragments

RNA

CTAB extraction

Ethidium bromide gel electrophoresis

Blue/white colony and T7/SP6 screening

DNA

Sequencing and sequence analysis

Cloning into pGEM-3zf vector

PCR using genus specific primer sets

RCA

Full length DNA fragments

Restriction digestion

Screening and detection scheme for plant virus groups in potato and vegetable samples collected from Iraq.

slide9

CI

NIaVPg

HC-Pro

NIa Pro

P1

P3

CP

NIb

~1.8 kb

~1.6 kb

  • Results:

5`UTR

Detection of potyviruses in Iraqi samples

3`UTR

Poly A Tail

M p To P Tomato Broad bean

Sample screening for potyviruses by Sprimer/M4T primer set. P: potato, To:tomato.

Zucchini leaf showing mosaic symptoms from naturally infected plant

slide10

Potato virus Y-O:N IRQ

99

PM2 IRQ

99

PM6 IRQ

PM9 IRQ

PM8 IRQ

98

PVY-Wi

99

PVYO:N

86

PB209 USA

N:O-Mb112 Canada

OR-1 USA

PVY

P55 IRQ

92

99%

P8-2 IRQ

P8-6 IRQ

96

SYR-II-DrH

PVYNW

P54A-3 IRQ

100

87

PVY-To IRQ

78

P46-1 IRQ

98

96

P54A-5 IRQ

P54A-1 IRQ

PVYNTN

PVY-NTN IRQ

100

PVY-12

96

HR1

Potato virus A

93

2002 Japan

Kunchyna Slovakia

98%

ZYMV

100

ZYMV IRQ

B strain Isael

100

AG Israel

NAT Israel

96

Gladiolus

BYMV-S

90-2 Japan

100

BYMV-IRQ1

BB5 IRQ

88

BYMV

BB3-IRQ

97%

BB17 IRQ

BB1-1 IRQ

100

BYMV-IRQ2

100

BB6 IRQ

BB15-IRQ

BB2 IRQ

BB3-1 IRQ

BB3-2 IRQ

Yam chlorotic necrotic mosaic virus

Blackberry virus Y

Squash vein yellowing virus

Brome streak mosaic virus

100

Oat necrotic mottle virus

100

Neighbor-Joining phylogenetic tree of potyviruses.

slide11

Detection of begomoviruses in Iraqi samples

ori

CR

AV2

AC4

DNA-A

V1 or CP

C2 or TrAP

C2 or REP

Naturally infected tomato plant showing yellowing and leaf curling symptoms

C3 or REn

~540 bp

~2.8 kb

L Tomato Eggplant +

L Tomato

Gel electrophoresis of DNA fragments amplified by PCR using Deng primers

RLFP of RCA products

slide12

New Caledonia

Moroccan

Almeria

97

RE4

89

95

Mauritius

Netherland

TYLCV IRQ

Jordan

100

Cuba

100

Dominican

Japan-Misumi

100

TYLCV-GJ

100

99

Bos Korea

100

Iran

Kuwait

94

TLCOV

TLCV-AU

TYLCV-China

TCTV

99%

TYLCV

Evolutionary relationships of Tomato yellow leaf curl virus isolated from tomato in Iraq to GenBank isolates/viruses.

slide13

ORF4

(TGB3)

25K

223K

7K

11K

ORF2

(TGB1)

ORF6

(NBP)

ORF1 (RRP)

ORF4(CP)

ORF3

(TGB2)

Poly A Tail

12K

34K

Detection of carlaviruses in Iraqi samples

~950 bp

Cowpea

P M

RT-PCR using Car-F1/M4T primer set to screen samples for carlaviruses. P:potato

slide14

P3-2

P3-4

82

P3-11

P3-11

P3-1

P3-1

P3-3

P3-12

P3-12

PMO2-cp

PMO2-cp

PVS3-5 Syria Syria

P19-18-cp

P19-18-cp

P19-23-cp

P19-23-cp

P19-24-cp

P19-24-cp

89

P18-33

P18-33

99

P18-34

P18-34

KER.SA.28 Iran

AZA.TA.6 Iran

P4-23

P4-23

PC11-cp

PC11-cp

Karla

Hangzhou China

China

IdDef-US

Id4106-US

90

PC6

Maranhao Brazil

91

Arachis Brazil

92

CpMMV-PR Puerto Rico

Barreiras Brazil

83

CPMMV-M USA

CPMMV-H USA

Ghana

Cp2-8

92

96

Cp2-9

99%

PVS

80%

CoMMV

Neighbor-Joining phylogenetic tree of carlaviruses.

slide15

33K

92K

22KD

ORF3

ORF4

41K

19K

~ 1.2Kb

M Tomato E

Tombusviruses RT-PCR using CIR1/CIR2 primers.

E: eggplant sample

Detection of tombusviruses in Iraqi samples

3`OH

5`

ORF1

ORF1 (RT)

ORF2 (CP)

Naturally infected eggplant showing leaf malformation symptoms

slide16

Tom9

Tom13

Tom10

Tom1

77

Tom5

Tom8

93

Tom11

Tom12

98

Tom6

GALV

Tom7

Japan

98

92

Limo 08

GYP2

92

Lim 4

99

WaterDoss

Lim 3

99

Italy

72

SchunterRiver

TBSV

94

CIRSV

CNV

CPMoV

100

LWSV

98%

Phylogenetic tree of Grapevine Algerian latent virus nucleotide sequences

slide17

Viruses detected in samples tested

PVY

PVS

GALV

PVY

TYLCV

GALV

ZYMV

BYMV

CpMMV

slide18

Conclusions

This study confirmed the occurrence of three potyviruses (PVY, ZYMV and BYMV), one begomovirus (TYLCV), two carlaviruses (PVS and CpMMV) and one tombusvirus (GALV) in potato and vegetables grown in Iraq.

Provided the first molecular information about plant viruses from Iraq and distinguished three recombinant strains of PVY; namely PVYNTN, PVYO:N and PVYNW occurring in potato and tomato samples.

The data presents a new report of CpMMV in cowpea and GALV in eggplant and tomato from Iraq.

Nucleotide sequence analysis revealed that all detected isolates showed high identity to GenBank isolates .

As Iraq imports most of its planting material, the high identity of sequences obtained to isolates from Genebank suggests that most viruses detected may have been introduced into Iraq in imported planting material.