CHARACTERISTICS OF BACTERIAL ACDS-GENE FROM THE STRAIN PSEUDOMONAS PUTIDA B-37 AND THE CREATION OF A GENETIC CONSTRUCT FOR DETERMINING ITS TRANSIENT EXPRESSION IN THE PLANT CELLS NICOTIANA BENTHAMIANA

Ethylene is an essential plant hormone also known as a stress hormone because its synthesis is accelerated by induction of a variety of biotic and abiotic stress. The plant growth promoting bacteria containing the enzyme 1-aminocyclopropane-1- carboxylate deaminase (ACC-deaminase) enhances plant growth by decreasing ethylene level under stress conditions. The expression of ACC-deaminase (acdS) gene in transgenic plants is an alternative approach to overcome the ethylene-induced stress. Agrobacterium-mediated DNA transfer is currently the most facile and versatile method to deliver gene constructs into the nucleus for gene function analysis in diverse plant species. Transient gene expression via Agrobacteriummediated DNA transfer in different plant tissues offers a simple and fast method to analyze transgene functions. In present work, the acdS-gene was amplified by PCR and then cloned into pBI121 vector under the control of the cauliflower mosaic virus (CaMV) 35s promoter. Agrobacterium tumefaciens GV3101 strain harboring pBI121-acdS vector jointly with the helper strain 19К were used for Agrobacterium-mediated leaf infiltration in Nicotiana benthamiana to infect 3-weeks-old plants. Monitoring of transient expression efficiency at 3 days post-infection was conducted by plant RNA extraction and RT-PCR. RNA was extracted from Nicotiana’s infiltrated zones and an amount of 1 μg total RNA was used to synthesize first-strand cDNA and then RT-PCR.

ACC-deaminase, acdS-gene, transient expression, Nicotiana benthamiana, Pseudomonas putida, Agrobaterium tumefaciens

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