Cauliflower mosaic Cauliflower mosaic virusvirus

Host RangeHost Range

►Narrow host range ,restricted Narrow host range ,restricted members of Brassicaceaemembers of Brassicaceae

►Vein clearin in Arabidopsis,Brassica Vein clearin in Arabidopsis,Brassica spe,Rhaphanus sps &other plants spe,Rhaphanus sps &other plants family in Brassicaceae family in Brassicaceae

ClassificationClassification

► The The CaulimoviridaeCaulimoviridae are a family of are a family of virusesviruses, including the , including the following genera:following genera:

► Genus Genus BadnavirusBadnavirus; type species: ; type species: Commelina yellow mottle virusCommelina yellow mottle virus

► Genus Genus CaulimovirusCaulimovirus; type species: ; type species: Cauliflower mosaic virusCauliflower mosaic virus

► Genus Genus TungrovirusTungrovirus; type species: ; type species: Rice tungro bacilliform virusRice tungro bacilliform virus

► Genus Genus SoymovirusSoymovirus; type species: ; type species: Soybean chlorotic mottle virusSoybean chlorotic mottle virus

► Genus Genus CavemovirusCavemovirus; type species: ; type species: Cassava vein mosaic Cassava vein mosaic virusvirus

► Genus Genus PetuvirusPetuvirus; type species: ; type species: Petunia vein clearing virusPetunia vein clearing virus

StructureStructure

► Camv is an icosahedral virus Camv is an icosahedral virus ► Diameter 50nm ,molecular weight 23*10(6) Diameter 50nm ,molecular weight 23*10(6)

daldal► genome is circular ds DNA,present between genome is circular ds DNA,present between ► 2 & 3 layers2 & 3 layers► Molecular weight 4.9*10(6)dalMolecular weight 4.9*10(6)dal► G+C content 43% G+C content 43% ► Capsid multilayeredCapsid multilayered► Arrangement of capsomere not clearArrangement of capsomere not clear

StructureStructure

                                            

   

       

                  

  

►The (-)strand :single discontinuity or The (-)strand :single discontinuity or gap gap

►The (+)strand:two gapsThe (+)strand:two gaps►Gaps are primer binding sites for Gaps are primer binding sites for

replication of DNA replication of DNA ►Gaps are same respect to specific Gaps are same respect to specific

restriction site restriction site

PropertiesProperties

►Thermal inactivation point 70 to 75 Thermal inactivation point 70 to 75 degree cell for 10 mintdegree cell for 10 mint

►DNA resistant to chloroformDNA resistant to chloroform► Infectivity weakened by treatment Infectivity weakened by treatment

with etherwith ether►Linear DNA not infectiveLinear DNA not infective►Serological studies:closely related to Serological studies:closely related to

dahila mosaic,straw berry banding and dahila mosaic,straw berry banding and carnation etched ring virusescarnation etched ring viruses

GenomeGenome

► Coding region I - Coding region I -  codes for 38 KD protein which  codes for 38 KD protein which helps in transfer of virus from one cell to another. helps in transfer of virus from one cell to another.

► Coding region II - Coding region II - codes for 19 KD protein which codes for 19 KD protein which attracts aphids. attracts aphids.

► Coding region III - Coding region III - codes for viral 15 KD viral codes for viral 15 KD viral capsomeres. capsomeres.

► Coding region IV - Coding region IV - codes for 57 KD coat protein. codes for 57 KD coat protein. ► Coding region V- Coding region V- codes for 79 KD reverse codes for 79 KD reverse

transcriptase transcriptase ► Coding region VI - Coding region VI - codes for 58 KD proteins which is codes for 58 KD proteins which is

used in building inclusion bodies. used in building inclusion bodies. ► Coding region VII - Coding region VII - codes for protein which targets codes for protein which targets

newly formed virus to inclusion body. newly formed virus to inclusion body.

ReplicationReplication

► Enry into the host cell by aphidsEnry into the host cell by aphids► Uncoating and release of DNA within Uncoating and release of DNA within

cytoplasmcytoplasm► Uncoated DNA transport to nucleusUncoated DNA transport to nucleus► Removal of overhang and gaps by DNAse Removal of overhang and gaps by DNAse

and DNAligaseand DNAligase► DNA associated with histones DNA associated with histones ► Two RNA transcripts produced and Two RNA transcripts produced and

transported to cytoplasm transported to cytoplasm

►35sRNA Codes structural proteins35sRNA Codes structural proteins►19sRNA involved replication19sRNA involved replication►dsDNA synthesised from RNAs by reverse dsDNA synthesised from RNAs by reverse

transcriptasetranscriptase► Encapsidation occurs form progeny Encapsidation occurs form progeny

virionvirion►Newly formed virus aggregate form Newly formed virus aggregate form

inclusion bodiesinclusion bodies►Mature virions escape from infected cell Mature virions escape from infected cell

symptomssymptoms

SymptomsSymptoms

►Vein clearing or chlorotic vein banding Vein clearing or chlorotic vein banding in in ArabidopsisArabidopsis thalianathaliana,,Brassica Brassica spe.spe.

►Mosaic symptoms occur in B. campestrisMosaic symptoms occur in B. campestris►Stunting of entire plantStunting of entire plant►Virus containing Inclusion bodies in the Virus containing Inclusion bodies in the

cytoplasmcytoplasm►Nucleus become lobedNucleus become lobed► vesicles protrude out from cell wallvesicles protrude out from cell wall

ControlControl

►Using insecticidesUsing insecticides►Elimination of disease plantsElimination of disease plants►Cultivation of virus –resistant varietiesCultivation of virus –resistant varieties►Selection of disease free planting Selection of disease free planting

materialsmaterials►Selection of disease free soil for Selection of disease free soil for

cultivationcultivation

APPLICATIONSAPPLICATIONS► The cauliflower mosaic virus (CaMV) 35S promoter has The cauliflower mosaic virus (CaMV) 35S promoter has

been most commonly used in plant transformation studies, been most commonly used in plant transformation studies, but its activity in mushrooms has not been reported.but its activity in mushrooms has not been reported.

► p301-b is a binary vector containing a bialaphos resistance p301-b is a binary vector containing a bialaphos resistance gene driven by the promoter of Lentinus edodes gene driven by the promoter of Lentinus edodes glyceraldehyde-3-phosphate dehydrogenase (GPD) gene. glyceraldehyde-3-phosphate dehydrogenase (GPD) gene.

► CaMV 35S-GUS was inserted into p301-b, and the resulting CaMV 35S-GUS was inserted into p301-b, and the resulting construct p301-bG was transformed to protoplasts of construct p301-bG was transformed to protoplasts of Ganoderma lucidum and basidiospores of Pleurotus Ganoderma lucidum and basidiospores of Pleurotus citrinopileatus. citrinopileatus.

► GUS activity was observed in the transformants, indicating GUS activity was observed in the transformants, indicating that CaMV 35S promoter can direct expression of that CaMV 35S promoter can direct expression of exogenous gene in the mushrooms. This is the first report exogenous gene in the mushrooms. This is the first report on the application of CaMV 35S promoter in genetic on the application of CaMV 35S promoter in genetic modification of mushrooms.modification of mushrooms.