Assessing Jatropha curcas pollen viability: A comparative assessment of transgenic and nontransgenic pollen under various environmental conditions using rapid staining technique”,

In Press
Frontiers in Plant Science

Kasthurirengan Sampath, Hong Yan and Srinivasan Ramachandran.

Pollen is the important vector for the transgene flow between non-transgenic plants. Several aspects of pollen biology have been investigated using pollens from X8#34 transgenic and non-transgenic Jatropha, a promising biofuel crop. Pollen viability was assessed through different staining methods and the optimized double staining method with Fluorescein Diacetate (FDA) and Propidium Iodide (PI) for Jatropha , as it can distinguishingly clearly differentiate viable and dead pollen with green and red fluorescent respectively.Among the environmental factors we tested, high temperature (35, 40 & 45°C), significant difference observed from 15 min incubation andperiod and high doses of UV-B irradiation significantly reduced Jatropha pollen viability at 12 and 15 W/m2 dosages. There is no significant difference in pollen viability between X8#34 transgenic Jatropha with non-transgenic counterpart. Under a field relevant sunny condition, pollen viability reduced to 19% in 45 min and 16% viability in non-transgenic, with complete loss of viability in 90 min for both. Under cloudy /shady atmospheric conditions, most of the Jatropha pollens (>97 %) lost their viability in 240 min. Pollen viability assessments showed that there is no statistical difference between two genotypes, X8#34 transgenic and non-transgenic Jatropha across all tested conditions. There is no significant difference in pollen viability of X8#34 transgenic Jatropha with their nontransgenic counterpart. This report is the first study on transgenic Jatropha curcas pollen viability study, we suggest that the optimized double staining technique (FDA+PI) in Jatropha and could serve bd adopted as the baseline for risk assessment of transgenic Jatropha plants.

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