Virus and Viroid As Plant Pathogens

(Agrios 2005, p 723 824)

Virus: Nucleoprotein(s) that can cause diseases in plants Submicroscopic entity that can reproduce in living host cells/tissues Viroid: Small (circular), low molecular weight RNA (naked) that can infect plant cells, replicate themselves, and cause disease Smaller than virus, 246 - 375 bp or nucleotides compared to 4 -20 kb of virus Prion: No nucleic acid, only proteins Diseases: scarpie disease in sheep bovine spongiform encephalopathy (mad cow) Creutzfeldt-Jacob disease in human

Important characteristics of virus

Obligate parasite One virus unit (virus particle) is called virion, that composed of 2 main components: Nucleic acid (RNA or DNA) Coat protein (capsid) Reproduce by replication, synthesize protein and nucleic acid

Virus Morphology
Elongated rod [rigid rods (15 x 300 nm) or flexious rod (10 - 13 nm wide x 480 - 2000 nm long] Short rod/cylindrical bacilluslike rod ~ 52-75 nm wide and 300-380 nm long Spherical [isometric or polyhedral) Majoriti: polyhedral, diameter: 17-60 nm
1 m (micron) = 1/1000 mm 1 nm (nanometer) = 1/1,000,000 mm

Virus morphology (cont.)

rigid rod

flexious rod

short rod

Isometric

Twin isometric

Viral Nucleic Acid

Importance: infectivity 2 types: single strand or double strand ssRNA/dsRNA or ssDNA/dsDNA Majority: ssRNA Component: 5-40% Long flexious rod: % nucleic acid low spherical: % nucleic acid higher

Protein coat (capsid)

Importance: Protect the nucleic acid Increase infectivity Content: 60-95%

Virus Replication
New Virion forms 10 hours after inoculation into plant cells Virus replication uses host enzymes and virus and host genetics Steps: Adsorption Penetration and release of nucleic acid from protein NA replication and biosynthesis of proteins assembly

History of Plant viruses and viroids

1886 1892 1898 1939 1971 1982 Mayer Iwanowski Beijerinck Kausche et al Diener Prusiner Tobacco mosaic symptom was induced in healthy tobacco Pathogens of tobacco mosaic was smaller than bacteria Tobacco mosaic was caused by contagious living fluid called virus Tobacco mosaic virus first seen with electron microscope Discovery of viroids in diseased potato Discovery of prions (animal diseases)]

Plant virus classification

Refer to Agrios pg 749 751

Kingdom: Viruses
A) Virus genera not yet assigned into families (+) ssRNA Rod shaped particles (1 ssRNA, 2 ssRNA, 3 ssRNA, 4 ssRNA) Filamentous particles (1 ssRNA) Isometric particles (1 ssRNA, 2 ssRNA) Bacilliform particles (3 ssRNA) (-) ssRNA Thin flexious multipartite viruses

Plant virus classification (cont.) B) Virus genera already assigned into families (+) ssRNA Filamentous particles (1 ssRNA, 1 or 2 ssRNA) Isometric particles (1 ssRNA, 2 ssRNA, 3 ssRNA) (-) ssRNA Bacilliform particles Membraneous circular particles dsRNA (+) ssDNA Isometric particles dsDNA Isometric particles Bacilliform particles ssRNA (RT) - retrotransposons

Symptoms of Diseases Caused by Phytopathogenic Viruses

1. Systemic symptom a) Mosaic b) Mottling c) Vein clearing d) Vein banding e) Chlorosis f) Vein reddening g) Witches broom h) Enation i) Leaf curl

2.

Local lesions ringspot Necrotic spot Chlorotic spot Other symptoms Stunting/Dwarfing Leaf rolling Yellowing/yellows

3.

Important plant virus diseases

Penyakit merah disease Host: padi Vector: green planthopper, Nephotettix virescens, Nephotettix nigropictus [Infective virus remains in vector for not more than 5 days] One Nephotettix can infect 40 plants/day Tungro (Philippines) Mentek (Indonesia) Leaves - yellowing, starting from the apex, twisted Stunted growth Decrease in yield 20-100%

Mosaic diseases
Chilli, tobacco, cucurbit, legumes/beans

Citrus tristeza
Symptom of vein-clearing on leaves Vector: brown citrus aphids, Toxoptera citricida Spread through grafting

Papaya ringspot
Ringspot on leaves/fruits Stunted growth / aphid transmitted

Cocoa swollen shoot (CSS)

In West Africa and Sri Lanka, absent in Asia Vector: koya (mealy bug, Planoccoides njalensis, P. citri)

Viroid Taxonomy
Depend on the presence or absence of a conserved central region 1. ASBVd or Avsunviroids Avsunviroideae Avsunviroid Pelamoviroid PSTVd or Pospiviroids Pospiviroideae: 5 subgroup: Pospiviroid, Apscaviroid, Cocadviroid, Coleviroid, Hostuviroid Unclassified Viroids

2.

3.

Important plant viroid diseases

Example of diseases Potato spindle tuber Cadang-cadang disease in coconut (Philippines) Citrus exocortis viroid

Transmission of plant viruses

Vegetative propagation (budding, grafting, organs like bulb, etc) Mechanical transmission through sap Seed transmission Pollen transmission (reduce fruit set, seed infection) Insect transmission (most common/important) Mite transmission Nematode transmission (Longidorus, Paralongidorus, Xiphinema) Fungus transmission Dodder transmission (parasitic plant, Cuscuta sp.)

Aphids

Thrip

Leafhopper

Whitefly

Insect Transmission
Non-persistent viruses (stylet-borne) Infective from few to several hours in vector, e.g., aphids Semi-persistent viruses Remain infective in vector from 1-4 days Circulative/persistent viruses Able to multiply inside vector (propagative viruses) Infective for longer periods Refer to Agrios p741-742

Translocation & Distribution in Plants

Movement from cell to cell through plasmodesmata Rate: 1 mm/day, 8-10 cells/day Leaf parenchyma cells Phloem transport Rate (faster): 15 cm/6 min Xylem transport

DETECTION OF PLANT VIRUSES

Inclusion bodies - p726 & 727 Transmission studies Grafting Budding Rubbing Dodder insect Purification and electron microscopy Serology - p 745/746 ELISA (enzyme linked immunosorbant assay) FAT (fluorescent antibody technique) Electrophoresis - p754 Use hybridization technique (cDNA probes) - p754 PCR techniques

IDENTIFICATION OF PLANT VIRUSES

Host range TIP (thermal inactivation point) LIV (longevity in vitro) DEP (dilution end point)