Plant Diseases Caused by Viruses and Prokaryotes

Mohamed Mannaa

Plant viruses
Like all other viruses, plant viruses are obligate intracellular parasites that do not have the molecular machinery to replicate without a host.

Viruses are extremely small and can only be observed with an electron microscope. The structure of a virus is given by its coat of proteins, which surround the viral genome. Viruses are placed into plant tissue by insects or vegetative propagation, or they enter through wounds. They usually become systemic within the host plant. An infected plant may be able to co-exist with the virus, or it may be killed. In any event, the plant cannot be cured of the viral infection.

 Viruses also cause many important plant diseases and are responsible for huge losses in crop production and quality in all parts of the world. Infected plants may show a range of symptoms depending on the disease but often there is leaf yellowing (either of the whole leaf or in a pattern of stripes or blotches), leaf distortion (e.g. curling) and/or other growth distortions (e.g. stunting of the whole plant, abnormalities in flower or fruit formation).

Yellow vein-banding symptoms on grapevine caused by Grapevine fanleaf virus.

Yellow mosaic symptoms on squash caused by Squash mosaic virus

The major means of control (depending on the disease) include:

Chemical or biological control of the vector (the organism transmitting the disease, often an insect) Growing resistant crop varieties: natural resistance and Transgenic resistance Use of virus-free planting material

Exclusion: the prevention of disease establishment in areas where it does not yet occur. This is a major objective of plant quarantine procedures

Bacteria
prokaryotic (no membraneenclosed nucleus) no mitochondria or chloroplasts a single chromosome, closed circle of double-stranded DNA If flagella are present, they are made of a single filament of the protein flagellin. The plasma membrane (in Gram-positive bacteria) and both membranes in Gram-negative bacteria are phospholipid bilayers

CROWN GALL
Caused by: Agrobacterium tumefasciens
SYMPTOMS: Rough abnormal galls develop on roots, crowns, and occasionally on aerial parts of stone fruit trees.
Young trees become stunted and older trees often develop secondary wood rots.

DISEASE CYCLE
The bacteria survive in gall tissue, in soil, or in apparently healthy root of host plants. The bacterium may persist in field soil for at least one year, or considerably longer if large amounts of infected root residues remain in soil after removal of trees. The pathogen penetrates only through wounds. The most susceptible wounds on stone fruits are the crown end roots and rarely on trunks and limbs. The bacteria attach to the wounded host cells and then transfer T-DNA into the cell. Infections are favored by moist , alkaline, poorly drained soils and can be stimulated by the feeding of plant parasitic nematodes. At temperatures above 20C galls become obvious in 2-4 weeks after infection.

DISEASE MANAGEMENT
Proper sanitation and cultural practices: the use of certified disease-free transplants, careful handling to avoid injury as much as possible during planting and life of trees in the orchard, and planting in well drained soils. Biological control, using a non pathogenic strain, A. radiobacter K84, that produces the antibiotic agrocin 84, is effective preventive treatment.

Phytoplasma
Phytoplasma is a prokaryote. wall-less intracellular bacteria. paleomorphic shape. cannot be grown in vitro (absolute parasite). multiplication by binary fission or budding. localized in the phloem sieve tubes of infected plants from where it is acquired by the vector for subsequent transmission. it invades systemically all plant organs.

Biology

Examples
Witches Broom Disease of citrus by Phytoplasma

Spiroplasma

Citrus Stubborn disease

Citrus Stubborn disease

Causal agent First mollicute of plant origin to be cultured (1970). Characterized as Spiroplasma citri in 1973.

Characteristics: Wall-less prokaryote Transmitted to plant phloem by leafhoppers Motility and helicity: cytoskeleton

(Charbonneau & Ghiorse, 1984)

(Yokomi et al., 2010)

GEOGRAPHICAL DISTRIBUTION of citrus stubborn disease

Based on EPPO Data Sheets
Europe France (Corsica only) Greece Italy (few records; Sardinia, Sicily) Spain Turkey

Asia Cyprus Iran Iraq Israel Jordan Lebanon

North America

Mexico USA (Arizona, California, Illinois, Maryland).

South America: (Unconfirmed)

Africa Algeria Egypt Libya Morocco Tunisia

Pakistan Saudi Arabia Syria Turkey Yemen

Argentina (Tucumn) Brazil (So Paulo) Peru Suriname Venezuela.

Oceania

New Zealand (isolated reports).

Develop best and show symptoms under hot conditions

Infect phloem sieve tubes

Acquiring S. citri from other hosts

S. citri multiplies in its vector, that become infective 10-20 days after acquisition

Symptoms
Tree symptoms:
Stunted growth Up-right foliation Shorter, chlorotic leaves. short internodes

Fruit symptoms:
Suppressed fruiting Lopsided, Acorn shaped. Styler end green. aborted seeds

Culturing

Biological indexing

Serological assay ELISA

Commercial kit (Sediag Italy)

washing

at

405 nm

Washing & Incubation

(AP-IgG)
(P-nitrophenylphosphate)

(Clark and Adams, 1977)

Washing & Incubation

Molecular assay
DNA extraction CTAB protocol
(Doyle and Doyle 1990)

PCR assay using P89 and P58

The targeted S.citri DNA

PCR assay using P32

Taq enzyme Primer

Designed on the putative adhesion gene P89 and the putative adhesion like multigene P58 Designed al, 2008) (Yokomi et on a plasmid gene, insect vector transmitted strains

PCR assay using Spiralin

Amplifi ed DNA

(Breton et Classical primer, al, 2010) designed on spriralin gene