<p>Pyramiding of AtHDG11 and Cry2Aa2 into pepper (Capsicum annuum L.) enhances drought tolerance and insect resistance without yield decrease</p><p>Supplemental Figure</p><p>Supplemental Fig. 1 a PCR analysis of T1 putative transgenic pepper plants: Lane 1, 2 Kb marker; lane 2, pCAMBIA1301-AtEDT1/HDG11-Cry2Aa2-PIM plasmid; lane 3, non-transgenic plant (WT); lanes 4–</p><p>15, randomly choosed 12 plants of T1 generation putative transformants used for result presentation, two bands of approximately 2100 bp and 1860 bp were obtained in the 10 progenies, while other 2 progenies without, indicate transgene had a co-segregation in T1 generation pepper plants. b RT-PCR analysis of</p><p>AtEDT1/HDG11 and Cry2Aa2 gene expression pattern in 4-week-old T1 transgenic lines, Actin serve as an internal standard. Lane 1, no cDNA template; lane 2, non-transgenic (WT) plant; line 3 T1 generation null transgene plant; lanes 4-6, T1 transgenic plants (T). Result indicated AtEDT1/HDG11 and Cry2Aa2 expressed in T1 transgenic lines, but not the in non-transgenic (WT) plant and T1 generation null transgene plant.</p><p>Supplemental Fig. 2 The fruit weight and components of transgenic (T) and non-transgenic (WT) pepper fruit under drought stress conditions. a Cross section of transgenic (T) and non-transgenic (WT) pepper fruit. Pepper fruit were picked from same relative position. Bar = 2 cm. b to d. Fruit length (b), fruit diameter (c), and fruit pericarp thickness (d) of transgenic (T) and non-transgenic (WT) pepper fruit.</p><p>1 Values are mean ± SD (n= 15, *p < 0.05, ** p < 0.01)</p><p>Supplemental Fig. 3 The cross section of transgenic (T) and non-transgenic (WT) pepper mature fruit. The cells in transgenic line (T) is larger than that of the non-transgenic (WT) line. Bar = 50 μm</p><p>- Supplemental Fig. 4 H2O2 content and O2 production rate of transgenic (T) and non-transgenic (WT) pepper plants under both normal (‘control’) and 6 d drought stress treatments. Values are mean ± SD of three independent experiments (** p < 0.01).</p><p>2</p>