Practical no : 6

Object : Non- persistence and semi-persistence transmission of viruses

Objective

 To know about the transmission of non-persistent and semi-persistent plant viruses

Material Required

 Vector aphids: must be aviruliferous

 Petri-dish

 Adhesive tapes to close dish

 Indicator plants: young, vigorously growing

 Filter paper

 Insecticide and sprayer

Theory

In non persistence transmission the virus is acquired by the vector after feeding on
parenchyma cell and epidermal cell. Probing takes as little as 5 seconds. Vector becomes
infective immediately after feeding, and virus lost by vector during molting. No latent
period, E.g CMV, BCMV etc.

In case of semi-persistence virus transmission, the virus is acquired from phloem after
long feeding, Persistence of virus in this vector for 10–100 hrs. No latent period is seen in
semi-persistence virus also. Virus do not circulate and multiply in its vector, Infectivity is
lost during moulting ,E.g. CaMV, CTV

Procedure for non-persistent transmission

 Disturb the colony of aviruliferous aphids in the host plant so that they withdraw
their stylet from feeding cell.
  This can be done by carefully tapping the abdomen with the brush, or by
breathing on them or by warming the leaf over an electric light bulb, otherwise
the fragile stylet will be damaged.
 Carefully pick up individual wingless aphids (at the rate of at least 5 aphids /
indicator plant), using the tip of the moistened paint brush and transfer to the
Petri-dish lined with moist (not wet) filter paper and close the dish with tape.
 Starve the aphid for at least 5 min to 1hr in a cool shaded place
 Cut the tape and transfer the aphids to infected plant material (to detached leaves
in a separate petri-dish if possible) to feed for about 2min and watch under
stereomicroscope whether the aphids are feeding.
 Transfer at least 5 aphids to each indicator plant as steps above
 Cover or place the plants in a cage and allow a transmission feed of 1 h.
 A variety of insect proof cages are found for IFP to avoid extraneous (coming
from outside) contamination.
 Kill the aphids with an insecticide, e.g. metasystox, by spraying or by dipping the
whole plant inside the solution and set the plants in glasshouse for growth at about
20 °C and symptom observation.

Procedure for semi-persistent transmission

 For semi-persistent viruses follow the same procedure but give an AFP of 4hr and
IFP of 1hr
 In all vector transmission tests, control tests also must be made to ensure that the
symptoms developed are not due to simply feeding by the vectors or due to
extraneous factors.
 In control test, the vectors should be fed on non-infected plants and transferred to
indicator plants giving the AFP and IFP as above.

Conclusion

Hence we were able to get knowledge about non persistence and semi-persistence
transmission of viruses.