Page 149 - Book of Abstracts

5 International Scientific Online Conference DOI: https://doi.org/10.15414/2021.9788055224015

th

TRANSIENT EXPRESSION OG UIDA GENE IN PHYSALIS PERUVIANA L. AND
PHYSALIS IXOCARPA BROTH.
Olha Yaroshko, Nadiia Matvieieva
Institute of Cell Biology and Genetic Engineering of the National Academy of Science of Ukraine, Kyiv,
Ukraine; E-mail.: 90tigeryaroshko90@gmail.com
Physalis peruviana L. and Physalis ixocarpa Broth. are representatives of the Solanaceae
Juss. family, native to Central America. Raw material obtained from these species is used in
medicine and agriculture.
The research aimed to confirm the possibility of transient expression of reporter uidA
gene in seedlings of P. peruviana and Likhtaryk variety of P. ixocarpa.
Physalis peruviana and cultivar Likhtaryk of Physalis ixocarpa was used in our research as
plant material. Seeds were sown in pots with soil and grown in greenhouse conditions (22–25
0 C, 14-hour light period, illumination – 3000–4500 lx). Ten day-old seedlings were infiltrated
with the Agrobacterium tumefaciens strain GV3101 carried pCBV19 genetic vector in the
vacuum chamber (for 5–10 min, 22–24 C).
0
We used two groups of the experimental plants treated with A. tumefaciens suspension.
The leaves of the first ones were cut with a scalpel before infiltration of bacteria for better
penetration of the agrobacterial suspension into the plant tissue. The leaves of the second group
of the plants were used without cutting. Leaves treated by the buffer instead of bacteria were
studied as a negative control.
Histochemical reaction for detection of β-glucuronidase activity was conducted at 37 C
0
th
according to Jefferson on the 10 day after infiltration of seedlings. Β-glucuronidase activity
was detected visually by the appearance of blue staining of the plant tissues. The plants of stable
transformed Nicotiana tabacum which carry in their tissues uidA gene were used as a positive
control.
The results of histochemical reaction of P. ixocarpa plants treated with A. tumefaciens
without previous scarification were positive for 30.5 % of the total quantity of infiltrated
seedlings. 94.6 % of plants infiltrated with agrobacterial suspension with the previous
scarification demonstrated a positive histochemical reaction. The results of histochemical
reaction of P. peruviana were positive for 14.3 % without previous scarification) and 16.7 %
(with the previous scarification). At the same time the groups of plants, which were used as
negative controls, did not turn blue after the histochemical reaction. The tobacco leaf surface
(positive control) was stained in a completely blue color.
The areas with blue staining were located mainly near midrib and occupied more than 60
% of the surface area of the leaf blade (in a group of plants, which were infiltrated with
Agrobacterium suspension and scarificated).
The possibility of transient expression of gus gene in seedlings of P. peruviana and P.
ixocarpa cv. Likhtaryk was confirmed. Obtained results show that seedlings of P. peruviana and
P. ixocarpa are susceptible to infection with A. tumefaciens and can be used for further
transformation experiments with selective, reporter and target genes.

Keywords: Physalis ixocarpa, Physalis peruviana, transient expression, uidA gene,
Agrobacterium tumefaciens.

Acknowledgments
The author is grateful to doctor Dz. Rakhmetov, who kindly donated the seeds of Physalis species. This
research didn’t obtain financial support or grant from funding agencies in the public, commercial, or not-
for-profit sectors.

5 International Scientific Conference Agrobiodiversity for Improving the Nutrition, Health, Quality of Life and |148
th
Spiritual Human Development
November 3 2021
rd