Stability of expression of Cry1Ac and Cry2Ab2 proteins in Bollgard-II hybrids at different stages of crop growth in different genotypes across cropping seasons and multiple geographies.

Bollgard-II cotton expressing Cry1Ac and Cry2Ab2 insecticidal proteins has been commercially cultivated in India since 2006 to control bollworms. These genes were introgressed into parental germplasm of numerous hybrids. Therefore, it is imperative that these insecticidal proteins are expressed in sufficient quantities in different tissues, throughout the season irrespective of genetic background or environmental conditions for effective performance. Here, we document results of a comprehensive study on pattern of expression of Bt proteins across different stages of crop growth in > 2000 cotton hybrids (Gossypium hirsutum), across 12 cropping seasons tested in the Northern, Southern or Central zones in India, in terminal leaf, pre-candle square and boll epicarp tissues. Statistical analysis of variability using Linear mixed effect model was used to estimate factors contributing to variability in expression of Bt proteins. For Cry1Ac, variability was maximally contributed by genotype × season × plant growth stage effect in terminal leaves and boll epicarp, while season effect drove variability in pre-candle square. In Cry2Ab2, season effect drove variability in three tissue types. Pre-candle square tissue had most variability in expression of both proteins followed by terminal leaf and boll epicarp. Further, expression of Bt proteins in 234 G. hirsutum × G. barbadense hybrids showed similar expression patterns as intra specific hybrids though there was a significant difference in expression levels. Cry2Ab2 was expressed in significantly higher amounts when genes were in homozygous state. Bt proteins were also found to be expressed in varied amounts in different tissues and were expressed even when hybrids were grown at sub-optimal temperatures.