presentation cmv summer course 2012

7/30/2019 Presentation Cmv Summer Course 2012

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GOOD MORNING...


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APPLICATION OF CUCUMBER MOSAIC VIRUS(CMV)MILD STRAIN TO CONTROL VIRULENT STRAIN OF CMV

IN CAYENNE PEPPER (Capsicum frutescensL.)

I Made Agus Dwi Ginartha*, I Dewa Nyoman Nyana**, I Gusti Ngurah AlitSusanta Wirya**, and Dewa Nyoman Delanata*

*Master Program on Agricultural Biotechnology Udayana University**Faculty of Agriculture, Udayana University


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ABSTRACT

Cucumber mosaic virus (CMV), belongs to the genus Cucumovirus, is one ofthe most serious plant pathogenic viruses with a broad host range includingboth monocotyledonous and dicotyledonous plants. Several studies showedthat the use of mild strains of certain viruses were able to protect the plantfrom the infection of virulent strains. This study was done to determine the

protective effect of several mild isolates of CMV (N1, N2, N3, N4) againstvirulent strain of CMV in the seedlings of cayenne pepper. Five treatments ofmild strains of CMV and control with five replications each were tested basedon randomized block design. Results showed that two mild strains namelyCMV-N2 and CMV-N4 were able to protect the cayenne pepper from theinfection of virulent CMV strain. The protective abilities of these two isolatewere 45.45% for CMV-N2 and 39.86% for CMV-N4. These results suggested

that these two strains of mild CMV might be used as vaccine candidates toprotect the cayenne pepper from infection of virulent CMV under fieldcondition.

Key words : cucumber mosaic virus, mild strain, protective effect, virulentstrain


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INTRODUCTION

Cayenne peppers have been widely produced in Bali for localconsumption but have been increasingly exported to the world market.

In Bali, cayenne peppers (C. frutescentL.) are usually affected by twosignificant different viral diseases having typical yellowing and mosaicsymptoms. The incidence of mosaic disease was more frequent mostly

caused by infection of CMV.

Mild strains of CMV have been reported have satellite RNAs (satRNAs)that is can influence the symptomps (Garcia-Arenal et al. , 2000).

Application of several mild strains of CMV (N1,N2,N3,N4) in cayenne

pepper seeds in this study is to analyzes the protective ability from eachmild strain. The best mild strain which have high value of protectiveability might be used as vaccine candidates to protect the cayennepepper from infection of virulent CMV under field condition.


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Materials and Methods

Inoculation of mild isolated (N1, N2, N3, N4) to cayenne pepperseedlings.

Inoculum prepared squash cayenne leaves mildsource of some virus isolates in

0.05 M phosphate buffer pH 7.0 (1:5 w / v) with a mortar and pestel. Mechanical

inoculation inoculum preparation done by rubbing with a cotton bud on the surface

of two leaves that have been fully developed and has been sprinkled celliteabrasive materials.


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ELISA Test (Enzyme-linked immunoassay)To confirmation of CMV mild strain infection in

cayenne pepper seddlings.

cayenne pepper samples from all unit

treatment were tested for CMV, using a double

antibody sandwich ELISA and indirect.


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Challange inoculationTo determine the protective ability of

each CMV mild strain isolates to control

virulent CMV. Twelve days after

inoculation, the first true leaves weremechanically inoculated with severe

strain of CMV isolate called challenge

inoculation.


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Symptoms in cayenne peppers

seedling (20 days)

N2 + S

Healthy seedling Mild Mozaic Severe Mozaic


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Treatments Plant Treatment of symptomatic

CMV Percentage (%)

CMV

concentration in

plants (ELISA test)

*

Height of

plant

Number of

leaf

Protective

effect (%)

20 days 30 days 40 days

M m M m M m

NT 0 0 0 0 0 0 - 31,8a 12,6a -

N1 0 40 0 20 0 40 + 25,8b 11,6b -

N2 0 20 0 20 0 60 + 30a 12,4a -

N3 0 20 0 40 0 40 + 26,8b 11,4b -

N4 0 40 0 40 0 60 + 29,4a 11,8a -

NT+S 100 0 100 0 100 0 +++ 19,6c 9c -

N1+S 20 20 40 60 60 40 +++ 23,8bc 11,2ab 22,38

N2+S 0 40 0 20 0 80 + 29,6ab 12ab 45,45

N3+S 20 20 40 60 40 20 +++ 25,8ab 11,4ab 30,06

N4+S 0 40 20 60 0 60 + 28,8ab 11,2ab 39,86

Table 1. The percentage of plants symptomatic CMV After inoculation N1, N2, N3, N4 and

Virus Threatening CMV-V3 (S)


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N4+S N3+S N2+S N1+S NT+S

The protective abilities of these two isolate were 45.45% for

CMV-N2 and 39.86% for CMV-N4.

Protective ability of mild strain to protect cayenne

pepper seedlings from virulent CMV


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CMV infection can cause a decrease in the rate of photosynthesis by

reducing the number khlorofil, increased respiration, increased enzyme

activity, decreases the amount of growth regulators can affect the functional

system of plant cells directly or indirectly. When the metabolic chaos can be

tolerated by the plants do not cause symptoms, while others have a

detrimental effect on the host, causing symptoms (Agrios, 1996).


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Conclusion

Two CMV mild isolates containing satRNA (-N2 and -N4)

successfully found to be having cross protection ability against CMV

severe wild isolate. These results suggested that these two strains

of mild CMV might be used as vaccine candidates to protect the

cayenne pepper from infection of virulent CMV under field condition.


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References

Garca-Arenal, F., and P. Palukaitis. 1999. Structure and

functional relationships of satellite RNAs of cucumber

mosaic virus. In Vogt P.K., Jackson A.O., eds. Satellites

and Defective Viral RNAs. Berlin, Springer-Verlag. 37

63.pp.

Agrios, G.N. 1996. Plant Pathology (in Indonesian

Language), Gajah Mada University Press.


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Thank You


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CONFIRM OF SatRNA ON SEEDLINGS

1 2 3 4 5 6 7 8 9 10 11 12 13 14

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SatRNA

presentation cmv summer course 2012

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