Dear Gürsel Karaca

Several factors affecting diseases development,

Pls. find the attached files, may answer your question, good luck

https://www.researchgate.net/publication/248150596_Lesion_development_factors_following_inoculation_of_Sclerotinia_sclerotiorum_mycelium_to_eggplant_tomato_and_French_bean

https://www.researchgate.net/publication/249446492_Sclerotinia_sclerotiorum_Epidemiological_Factors_Affecting_Infection_of_Greenhouse_Aubergine_Crops

http://link.springer.com/article/10.1007/BF02979548#page-1

http://site.iugaza.edu.ps/tbashiti/files/2011/02/sdarticle113.pdf

http://synapse.koreamed.org/Synapse/Data/PDFData/0184MB/mb-31-113.pdf

http://apsjournals.apsnet.org/doi/abs/10.1094/PDIS.2004.88.8.891

http://www.scielo.br/pdf/hb/v29n1/21.pdf

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3988091/

Regards

Prof. Houda Kawas

Article Lesion development factors following inoculation of Scleroti...

Article Sclerotinia sclerotiorum: Epidemiological Factors Affecting ...

I already had most of the papers. I'll try inoculation once more both on the stem and on the lower leaf.. Thanks a lot.

Hi , the best way to get infection with Sclerotinia spp on the field , just spread the sclerotia in the field between the plant and the infection from ascospore, the infection will start after few week,

Yes multiply the inoculum of Sclerotinia spp on sorghum grains containg 0.2% honey and after good growth is observed apply @50 gm per sq. meter.

There are several techniques of inoculating sclerotia in stem, while conducting my phd research work and supervising post graduate thesis projects i used tooth pick inoculation technique i can send you 

PREPARATION OF TOOTH PICKS

Six centimeter bamboo wood toothpick  having pointed ends were soaked in water for 12 hrs. These toothpicks were softened and making free from wood chemicals by boiling for 4-5 times in fresh water for one hour till the water remained colourless on boiling which indicated that tooth picks have become free of wood chemicals. These chemical free toothpicks were spread on blotter sheet and air dried under shade for 24 hrs. The toothpicks were then transferred to autoclavable flasks 1/3rd filled with Potato Dextrose Broth (PDB). No loose spacing was left among the toothpicks in the flasks.  The flasks were autoclaved at 15psi, at 120°C for one hour and then cooled down to room temperature. It was then inoculated with a 5mm plug from the periphery of actively growing 7 days old test culture of Macrophomina phaseolina and placed at the tip of the bundle of toothpick. The flasks were incubated at 30°+ 2°C for three weeks. Forty eight field isolates  were inoculated separately with each of the test culture. During incubation period for maintaining the aerobic conditions screw of the flasks were slightly loosed after every 2-3 days interval. During this incubation period, toothpicks were covered with mycelia and sclerotia mass of M.  phaseolina.

 i have inoculated the  apical meristems using eggplants of one month growth stage , with PDA mycelial pathogen plugs (5mm)  of sclerotinia   .The results shown symptoms developed within 8 dayse after inoculation  in green house conditions .

I hope you had plnpriked  the tissue before inoculation? 

You can also try the suspension of fungus spore that are sprayed on the plants.

For the entry of the pathogen an injury of the host is important 

USE AGAR PLUG, BUT YOU NEED WOUND THE  HOST TISSUES AND COVER THE INCULUM WITH AUTOCLAVATED  MOIST COTTON  AND PARAFILM