TY - JOUR

T1 - Characterization of Transgenic Kalanchoë and Petunia with Organ-Specific Expression of GUS or GA2 ox Genes Led by the Deletion BOX-I Version (dBI) of the PAL1 Promoter

AU - Gargul, Joanna Maria

AU - Mibus-Schoppe, Heiko

AU - Serek, Margrethe

N1 - Publisher Copyright: © 2017, Springer Science+Business Media New York. Copyright: Copyright 2017 Elsevier B.V., All rights reserved.

PY - 2017/6/1

Y1 - 2017/6/1

N2 - In the present work, transgenic Kalanchoë blossfeldiana and Petunia hybrida with overexpression of Nicotiana GA2ox inserted in pCAMBIA1303 T-DNA are investigated. To avoid possible adverse effects of constitutive overexpression a modified Pisum PAL1 promoter was used, in which the BOX-I AC-rich motif had been deleted (dBI—deletion BOX-I). To investigate the tissue-specificity of the dBI and PAL1 promoters, their sequences were fused with the GUS gene, cloned into two types of vectors (pCAMBIA1303 and p6N) and introduced by Agrobacterium-mediated transformation into both species. GUS-transgenic lines were tested for the GUS mRNA in stems, leaves, and roots with the use of RT-PCR and GUS-stained tissues were visualized and compared with the use of light microscopy. The dBI promoter leads to expression of GUS mRNA and GUS activity in stems and petioles but not in roots or leaves, whereas the PAL1 promoter is less specific. All transgenic lines were tested for transgene copy number using Southern blot analysis. Transgenic Kalanchoë plants with Nicotiana GA2ox exhibited more than twofold reduction in stem length but no reduction of the number of internodes. Similarly, in Petunia transgenic plants, the stem length was reduced threefold. The leaves of the transgenic Kalanchoë plants were smaller, as convex as those of the youngest leaves of the non-transgenic control plants, and had reduced petiole length. The leaves from transgenic Petunia plants were similar in shape to those of the non-transgenic control plants but significantly smaller.

AB - In the present work, transgenic Kalanchoë blossfeldiana and Petunia hybrida with overexpression of Nicotiana GA2ox inserted in pCAMBIA1303 T-DNA are investigated. To avoid possible adverse effects of constitutive overexpression a modified Pisum PAL1 promoter was used, in which the BOX-I AC-rich motif had been deleted (dBI—deletion BOX-I). To investigate the tissue-specificity of the dBI and PAL1 promoters, their sequences were fused with the GUS gene, cloned into two types of vectors (pCAMBIA1303 and p6N) and introduced by Agrobacterium-mediated transformation into both species. GUS-transgenic lines were tested for the GUS mRNA in stems, leaves, and roots with the use of RT-PCR and GUS-stained tissues were visualized and compared with the use of light microscopy. The dBI promoter leads to expression of GUS mRNA and GUS activity in stems and petioles but not in roots or leaves, whereas the PAL1 promoter is less specific. All transgenic lines were tested for transgene copy number using Southern blot analysis. Transgenic Kalanchoë plants with Nicotiana GA2ox exhibited more than twofold reduction in stem length but no reduction of the number of internodes. Similarly, in Petunia transgenic plants, the stem length was reduced threefold. The leaves of the transgenic Kalanchoë plants were smaller, as convex as those of the youngest leaves of the non-transgenic control plants, and had reduced petiole length. The leaves from transgenic Petunia plants were similar in shape to those of the non-transgenic control plants but significantly smaller.

KW - 35S promoter

KW - Compact growth

KW - GAox

KW - Kalanchoë

KW - Petunia

KW - Stem- and petiole-specific promoter

UR - http://www.scopus.com/inward/record.url?scp=85009883066&partnerID=8YFLogxK

U2 - 10.1007/s00344-016-9650-x

DO - 10.1007/s00344-016-9650-x

M3 - Article

AN - SCOPUS:85009883066

VL - 36

SP - 424

EP - 435

JO - Journal of Plant Growth Regulation

JF - Journal of Plant Growth Regulation

SN - 0721-7595

IS - 2

ER -