Plant materials and growth conditions
The rice cultivar Nip, rice blast susceptible cultivar LTH,Pik-H4 NILs in LTH background were used for constructing
transgenic plants and inoculation assays. TheRFP ::PPik-H4 ::GFP /Nip,
PPik1-H4 ::GUS/Nip and
PPik2-H4 ::GUS/Nip were used for expression
pattern analysis of Pik-H4 . The
OE-Pik1-H4 -GFP /Pik and
OE-Pik2-H4 -GFP /Pik were generated
by crossing OE-Pik1-H4 -GFP /LTH and
OE-Pik2-H4 -GFP /LTH with Pik-H4 NILs
respectively. Nip was also used for protoplast transformation to
identify bidirectional promoter activity. Nicotiana benthamianawas used for agroinfiltration to evaluate promoter activity and HR
assays.
Rice cultivars grew on soil in a greenhouse with a natural light cycle
and 30°C/light, 26°C/dark. Tobacco seeds were germinated in MS medium
and transferred to the soil after germination in a growth chamber with a
16-h/8-h light/dark cycle at 24°C.