Reference "Marker-free transgenic (MFT) near-isogenic introgression lines (NIILs) of 'golden' indica rice (cv. IR64) with accumulation of provitamin A in the endosperm tissue"

Reference ID

26128

Title

Marker-free transgenic (MFT) near-isogenic introgression lines (NIILs) of 'golden' indica rice (cv. IR64) with accumulation of provitamin A in the endosperm tissue

Source

Plant Biotechnol J, 2006, vol. 4, pp. 467-475

Authors (9)

Baisakh-N	Rehana-S
Rai-M	Oliva-N
Tan-J	Mackill-D-J
Khush-G-S	Datta-K
Datta-S-K

Abstract

We have developed near-isogenic introgression lines (NIILs) of an elite indica
rice cultivar (IR64) with the genes for beta-carotene biosynthesis from
dihaploid (DH) derivatives of golden japonica rice (cv. T309). A careful
analysis of the DH lines indicated the integration of the genes of interest
[phytoene synthase (psy) and phytoene desaturase (crtI)] and the selectable
marker gene (hygromycin phosphotransferase, hph) in two unlinked loci. During
subsequent crossing, progenies could be obtained carrying only the locus with
psy and crtI, which was segregated independently from the locus containing the
hph gene during meiotic segregation. The NIILs (BC(2)F(2)) showed maximum
similarity with the recurrent parent cultivar IR64. Further, progenies of two
NIILs were devoid of any fragments beyond the left or right border, including
the chloramphenicol acetyltransferase (cat) antibiotic resistance gene of the
transformation vector. Spectrophotometric readings showed the accumulation of up
to 1.06 microg total carotenoids, including beta-carotene, in 1 g of the
endosperm. The accumulation of beta-carotene was also evident from the clearly
visible yellow colour of the polished seeds.