ASAD Thesis Defence

8/8/2019 ASAD Thesis Defence

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INVESTIGATIONS ON SUGARCANE MOSAIC VIRUS IN PUNJAB AND NWFP:

CHARACTERIZATION AND IDENTIFICATION OF RESISTANCE SOURCES


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PRESENTATION BY

MUHAMMAD ASAD FAROOQ

SUPERVISOR DR. ABID RIAZCO-SUPERVISOR DR. TAHIRA YASMIN

MEMBER DR. S. M. MUGHAL

MEMBER HUMAYUN JAVED

SUPERVISORY COMMITTEE


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INTRODUCTION


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Sugarcane -- an important cash crop of

the world-- accounts for over two third

of world sugar production

In Pakistan -- sugarcane is a high valuecash crop.

-- mainly grown for production

of essential item forindustries like sugar, chipboard,

and paper etc.


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Share in -- Value added of agriculture: 3.6 %

-- GDP: 0.8 % (Anonymous, 2010).

World sugarcane yield is limited by various diseasescaused by -- fungi, bacteria, virus and phytoplasma.

A significant share of yield losses is attributed to:

VIRUSES and PHYTOPLASMAThe most important among them is:

Sugarcane Mosaic Virus (SCMV)

Belongs to family POTYVIRIDAEOccurs -- in almost all the sugarcane growing

countries, except Mauritius and Guyana

(Viswanathan and Mohanraj, 2001).


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OBJECTIVES

To conduct quantitative surveys of Punjab and NWFP

to study occurrence, prevalence and distribution ofSCMV.

To identify weed hosts of SCMV.

Identification and characterization of SCMV through: Symptomatology

Differential Hosts

Serology (DAS-ELISA) Screening of sugarcane germplasm against SCMV

under inoculated conditions to identify/find sources of

resistance.


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MATERIALS AND METHODS


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The study was conducted in the Department ofPlant Pathology, PMAS-Arid Agriculture

University, Rawalpindi in collaboration with Crop

Disease Research Program and Sugar Crops

Research Program of National Agricultural

Research Center, Islamabad during the year

2008-09.


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FIELD SURVEYS AND COLLECTION OF SAMPLES

Data were collected regarding: Percent Disease Incidence Relative Occurrence and

Varietal Reaction

Punjab NWFP

Mandi Bahauddin MardanSargodha Peshawar

Faisalabad Charsadda

Jhang Dargai

Toba Tek SinghOkara

Rahim Yar Khan

Bahawalpur

MuzaffargarhLayyah


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Percent Disease Incidence:

An area of 10-X-10 m was selected --- all plants were counted

No. of infected plants% D.I. = --------------------------------------- x 100

Total No. of plants observed

Relative Occurrence (%):

Samples -- Whole leaves with characteristic symptoms of SCMV -- from the topof the plants

Kept in sterilized polythene bags -- Placed in a refrigerator at 4 0C untilprocessed.

Tested to confirm the presence of SCMV through DAS-ELISA

The relative occurrence was determined by applying following formula;

Number of samples positive for a Virus

Relative Occurrence (%) = ------------------------------------------------------ x 100

Total number of samples tested


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MAINTENANCE OF VIRUS ISOLATES

ELISA positive samples were stored out

Virus from samples having maximum

concentrations were multiplied on virussusceptible sugarcane varieties-- through

mechanical inoculation besides glass house

conditions for further studies.


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INDEXING OF SAMPLES THROUGH ELISA

Samples -- both provinces -- testedthrough DAS-ELISA standard buffersand protocol.

The samples were replicated twice.

The optical density -- measured at 405 nm-- ELISA plate reader -- with continuousmovement absorbance mode.


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IDENTIFICATION OF WEED HOSTS

Weeds -- from sugarcane fields -- carrying SCMV.

Record of local/vernacular and botanicals names was

maintained.

All samples were kept in sterilized polythene bags --

placed in a refrigerator at 4 0C until processed.

Samples were then tested to confirm the presence ofSCMV through DAS-ELISA.


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DIFFERENTIAL HOST STUDIESDifferential hosts tested: wheat, barley, maize, millet andsorghum.

Seeds were obtained from Crop Sciences Institute,NARC, Islamabad.

Test plants were grown in 4 clay pots (9dia) -- filled with

a mixture of field soil, sand and FYM (1:1:1) -- underglasshouse conditions -- in an insect free environment.

Plants in 3 clay pots -- 3 to 4 leaves stage -- weremechanically inoculated -- a mixture of representative

isolates -- Punjab, NWFP and Islamabad.The plants were observed for symptom appearanceweekly 25-30 days after inoculation the young growingleaves were collected -- tested through ELISA to confirm

the presence or absence of SCMV.


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SCREENING OF SUGARCANE GERMPLASMGermplasm lines of diverse sources being maintained at NARC.

Hot water treatment at 55 oC for 10 minutes.

These lines were mechanically inoculated with SCMV at 3-4leaves stage.

The plants were observed for symptom appearance weekly.

25-30 days after inoculation -- young growing leaves werecollected -- tested through ELISA to confirm the presence orabsence of SCMV.

The lines thus tested were grouped as resistant -- moderatelyresistant -- moderately susceptible -- susceptible on the basis ofdisease rating scale of Piperet al., (1996).


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SCALE USED FOR RATING SEVERITY OF SCMV ON SEEDLING

Rating Description of symptoms

0 Healthy, no virus visible.

1 Very mild symptoms on one or more leaves.

2 Mild symptoms on one or more leaves.

3 Moderate symptoms on one or more leaves.

4 Moderate symptoms (as in 2 or 3) but more widespread than 3.

5 Severe symptoms, widespread on plant. Especially, to all leaves on a tiller showingsymptoms.

6 Severe symptoms, as in 5, but in addition either noticeable stunting or small tomoderate amount of necrosis.

7 Very severe symptoms, severe stunting, obvious and significant amount of necrosis.

Where,

0 = - = Resistant (R)

1-2 = + = Moderately Resistant (MR)

3-4 = ++ = Moderately Susceptible (MS)

5-7 = +++ = Susceptible (S)


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RESULTS AND DISCUSSION


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Disease Incidence (percent) of SCMV in different districts of Punjab on

the basis of symptom observation

S. No. Name of District Mean D.I. (%) of two locations

of each district1. Mandi Bahauddin 41.89

2. Sargodha 70.46

3. Toba Tek Singh 51.12

4. Jhang 32.74

5. Faisalabad 15.95

6. Okara 75.53

7. Bahawalpur 35.87

8. Rahim Yar Khan 45.75

9. Muzaffargarh 34.34

10. Layyah 33.81


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Disease Incidence (percent) of SCMV in different districts of NWFP on the

basis of symptom observation

S. No. Name of District D.I. (percent) Mean of two locationsof each district

1. Peshawar 43.16

2. Mardan 53.03

3. Dargai 52.71

4. Charsadda 52.13

Relative occurrence (%) of random and non randomly collected samples from Punjab


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0

10

20

30

40

50

60

70

Ma

ndiB

ahauddin

Sarg

odha

T

obaTe

kSingh

Jhan

g

Faisa

laba

d

Oka

ra

Baha

walp

ur

Rahim

Yar

Kha

n

Muz

affarg

arh

Layy

ah

% R.O. (Random) % R.O. (Non Random)

Relative occurrence (%) of random and non-randomly collected samples from Punjab


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0

5

10

15

20

25

30

35

40

45

50

Pesha

war

Mardan

Charsadda

Dargai

% R.O. (Random) % R.O. (Non Random)

Relative occurrence (%) of random and non-randomly collected samples from NWFP

Response of different cultivars to SCMV in different districts of Punjab


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S. No. Name of District Locations Name of Variety Location vise D. I. (%)

1. Mandi Bahauddin Field 1 HSF-240 14.11

Field 2 Kala India 69.68

2. Sargodha Field 1 Unknown 90.26

Field 2 CPF-238 50.67

3. Toba Tek Singh Field 1 CPF-237 21.50

Field 2 China 80.75

4. Jhang Field 1 SPSG-79 35.94

Field 2 NSG-555 29.55

5. Faisalabad Field 1 HSF-242 19.01

Field 2 HSF-240 12.90

6. Okara Field 1 CP-90 71.93

Field 2 Mixture 79.14

7. Bahawalpur Field 1 CPF-234 35.95

Field 2 CPF-234 35.79

8. Rahim Yar Khan Field 1 CPF-234 44.64

Field 2 CPF-234 46.86

9. Muzaffargarh Field 1 HSF-242 29.53

Field 2 Desi + Mixture 39.15

10. Layyah Field 1 CPF-238 46.22Field 2 CPF-234 21.40

Response of different cultivars to SCMV in different districts of Punjab

Response of different cultivars to SCMV in different districts of NWFP


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S. No. Name of District Locations Name of Variety Location vise D. I. (%)

1. Peshawar Field 1 CP-77/400 41.71

Field 2 Sanober 44.60

2. Mardan Field 1 CP-77/400 62.44

Field 2 CP-77/400 43.62

3. Dargai Field 1 CP-77/400 52.94

Field 2 CP-77/400 52.48

4. Charsadda Field 1 CP-77/400 60.76

Field 2 CP-77/400 43.49

Response of different cultivars to SCMV in different districts of NWFP


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Different weeds identified as natural hosts of SCMV

S. No. Local Names Scientific Names of Weeds OD405

Value Reaction againstSCMV

1. Baroo Sorghum halepense Pers. 0.439 Positive

2. Bathoo Chenopodium album L. 0.129 Negative

3. Bhakra Tribulus terristris L. 0.119 Negative

4. Bhoin Cyperus iria L. 0.121 Negative

5. Damb Grass Polypogon monspeliensis Desf. 0.395 Positive

6. Deela Scirpus maritimum L. 0.207 Negative

7. Ghoin Cyprus difformis L. 0.112 Negative8. It-sit Trianthema portulacastrum L. 0.126 Negative

9. Kara Bara Digitaria adscendens L. 0.108 Negative

10. Khabbal Cynodon dactylon (L.) Pres. 0.328 Positive

11. Lelhi Convolvulus arvensis L. 0.119 Negative

12. Loomar Grass Setaria pumila (Poir.) Roem & Schult. 0.304 Positive

13. Lumb Grass Leptocloa panacea (Retz.) Ohwi. 0.459 Positive

14. Madhana Dactyloctenium aegyptium (L.) Willd. 0.349 Positive

15. Moorak Cyperus rotundus L. 0.127 Negative

16. Naroo Paspalum distichum L. 0.411 Positive

17. Swanki Echinochloa crusgalli (L.) Beauv 0.512 Positive


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Reaction of different host species on inoculation with SCMV

Host plant Cultivars Symptom Observations DAS-ELISA(SCMV)

Wheat Chakwal-97 No visible symptoms -

NARC No. 1542 No visible symptoms -

Barani-73 No visible symptoms -

Sorghum Js-62 No visible symptoms -

Millet MB-87 Light yellowing & mosaic +

Super-1 Light mosaic +

Acc. No. 8808 Light yellowing & mosaic +

Oats PD2LV

65No visible symptoms +

Maize EU-1097 Mosaic +

Rakaposhi Mosaic (7-8 leaf stage) +

Islamabad Gold Mosaic +

Islamabad white Mosaic +

Sweet Corn Mosaic and light yellowing +

Barley (Soorab-96) No visible symptoms -


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Reaction against SCMV Name of Varieties

Resistant (22) HSF-240, BF-129, CP-48-103, CPM-13, CO-1321,SPF-213, M-93, CP-43-33, M-92, CPF-237, NIA-98,SPF-234, Thatta-10, BF-162, COL-54, LRK-2001,BL-4, HSF-242, L-113, CP-51-21, L-118 and CP-44-107

Moderately Resistant (04) COJ-84, SPSG-26, PR- 1000 and L-116

Moderately Susceptible (06) CP-72-2086, Triton, CP-77-400, IM-61, L-357 andL-62-96

0 = - = Resistant (R)1-2 = + = Moderately Resistant (MR)

3-4 = ++ = Moderately Susceptible (MS)

5-7 = +++ = Susceptible (S)

Resistance potential of commercial sugarcane varieties against SCMV


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Reaction against SCMV Name of Germplasm Lines

Resistant (38) TCP-88-3480, CO-436, THATTA-8, PR-66-1240, SPHS-8, PR-68-123,

HSTH-18, CP-88-1165, CPTH-1, HSTH-10, HSTH-9, TCP-85-1432, TCP-69-1059, COJ-76, COJ-64, CO-312, CPTH-19, CP-81-1238, RB-72-454,CP-88-1540, L-89-152, CPHS-3, SPHS-2, HS-12, RB-82-5336, CPTH-20,CP-50-28, CP-88-1573, CP-70-321, HOSG-449, S-00-SPSG-1607, S-01-HOSG-795, S-02-HOSG-155, HOCP-92-631, N-60, Malakand-7, SPF-213and CP-84-1198

Moderately Resistant (10) RB-78-5148, GT-1, NCO-310, L-89-113, TCP-81-3067, S-98-CSSG-668,S-03-CPSG-433, S-03-HOSG-1275, S-03-HOSG-945 and S-97-US-183

Moderately Susceptible (15) CPF-150, AEC-81-89, CP-76-331, CPF-222, GT-11, CO-285, TCP-83-3211, COJ-79, CP-90-951, COJ-78, TCP-83-3210, HOSG-104, HS-4, S-98-SP-729 and CP-85-1382

Susceptible (13) CP-73-1030, CPTH-16, CPTH-3, TCP-86-3368, TCP-81-10, CPM-76-611,MEX-57-473, SPSG-93, HOCP-91-552, HOCP-91-559, CP-89-2376, CP-85-1491 and CP-89-1945

Resistance potential of sugarcane germplasm linesagainst SCMV

0 = - = Resistant (R)

1-2 = + = Moderately Resistant (MR)

3-4 = ++ = Moderately Susceptible (MS)5-7 = +++ = Susceptible (S)


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CONCLUSION

The sugarcane fields of major cane growing districts of Punjab and NWFP


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The sugarcane fields of major cane growing districts of Punjab and NWFPwere found overwhelmingly infected with SCMV.

SCMV was prevalent in all sugarcane-growing districts of Punjab andNWFP with the maximum disease incidence in Okara and Mardan

respectively.

The relative occurrence of SCMV was found highest in Muzaffargarh districtof Punjab, while it was highest Peshawar district of NWFP.

Eight weed species were recognized as natural hosts of SCMV and wereamong the dominant weed flora of both provinces.

Common maize and millet cultivars may also harbor SCMV in naturalconditions.

Disease may spread rapidly when vector population, particularly of Cornleaf aphid (Rhopalosiphum maidis) increases in the maize fieldssurrounding the fields of sugarcane, a commonly observed situation inPunjab and NWFP.

Out of 32 commercial sugarcane cultivars evaluated for their resistance

against SCMV, four were found moderately resistant and six were foundmoderatel susce tible to SCMV.

C lti ti f i d i l i ti ith k t ti l i t


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Cultivation of improved commercial varieties with known potential againstSCMV is therefore recommended so that, unaddressed losses due to thisdisease may be minimized.

Ten sugarcane advance lines were found moderately resistant, fifteen lines

were found moderately susceptible and only thirteen lines were foundsusceptible to SCMV.

Preferred parameters are quantitative i.e. cane weight, yield, germination,vigor & sugar contents, rather than qualitative i.e. resistance against pests ordiseases, drought tolerance, frost tolerance etc.

This results in failure of varieties due to low level of resistance againstdiseases especially of viruses. Therefore the lines identified as completely orat least partially resistant to SCMV must be included in yield trials as so thatresistant varieties with good yield potential may be put forward to the farmers

Week crop stand due to unavailability of required inputs well on time, poorphytosanitary measures and meager check on insect pests worsen thesituation collectively favor the virus to establish and become inhabitant inpotential areas as poor crop stand, higher weed infestation and large vectorpopulation play a role that is more favorable for SCMV to become sowidespread in nature.


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