Abstract:
Genetic transformation of two gene constructs 11GIy-1 and PsCIPK (that caIn: p131121 of
14KDa plasmid with /i-giucuronidase p35SGLSINT and plain selectable marker gene
npttl) confer both salt and drought tolerance to mesta variew t-IS-24 (hibiscus
sahi/iviffli) has been undertaken in the Genetic Lingineering Laboratory of Genetic
Resources and Seed Division, fliRt. Dhaka-1207. Cotyledons (with attached petioles)
and embryos were used as explants br indirect regeneration ob transformed plants and
direct transihrmation consecutively. GUS assay was perlormed alter 30 days of cocultivation
of infected Cotyledons. GUS positive putative blue color was developed up to
76% ( in case of PsCIPK gene) of cotyledons. But it was observed that 1-15-24 variety
was mild transtbrmed (32% (1115 blue color revealed) with PGIy-1 gene construct
(cotyledons). GUS positive explants (.Cotyledons) were cultured on shoot regeneration
medium (MS medium supplemented with IAA 0.5 mg/I and RAP 2 mg/I) with
cefotaxime (500 mg/I). Shoot regeneration percentage oftransfornied plants was 42 to 47
% depending on gene constructs. Transgenic HThLccus .va/ckiri//, plants have also been
produced by direct transformation procedure. Embryo axes ol mature seeds were infected
by immersion in a suspension of Agmhacicrnun strain culture Following a 24 h cocultivation
with .1g/?backhuin strain, embryos were grown on MS medium added with
a killer concentration of kananiyein (50 mg/mI) during 4 weeks at room conditions and
thereafter translèrred to greenhouse. 76.67% to 83.3410* of the explants excised from
putative translbrmed plants were found to be GUS positive depending on gene
constructs For salt tolerance testing, \'IS media supplemented with NaCl of different
concentrations (50, 75, 100 and 125 nN1) were used. respectively. The results showed
the genetic transformation of salt tolerance genes in I 15-24 variety and indicated the
possibility of the development oftransgenic niesta (Hibiscus .cahcicui/iz) varieties against
salt stresses. From this experiment, the tremendous observation was that Hibiscus
scthc/ari/a showed strong response to PsCIPK gene construct but mild response to PGly-
I gene construct We also reported here a rapid and efficient method for producing
uansgcnic plants rather than regeneration method from H. scthcktriffii embryos, which did
not require in ri/lu regeneration.
Description:
A Thesis
Submitted to the Faculty of Agriculture,
Sher-e-Bangla Agricultural University, Dhaka,
in partial fulfillment of the requirements
for the degree of
MASTER OF SCIENCE IN GENETICS AND PLANT BREEDING
SEMESTER: Jul-Dec’ 2009
