Screening Robinia pseudoacacia L. seedlings for salt stress resistance for subsequent detection of resistant genotypes by molecular genetic methods

The aim of this work was development of a screening system to find salt stress tolerant black locust seedling for following identification of stress tolerant genotypes. Our system combined original protocol of black locust seed germination under salt stress conditions and originally modified DNA extraction protocol. Salt stress induced changes in germination capacity, morphological indexes and biochemical markers confirmed stress development in proposed screening system. Used DNA extraction method allowed preparing DNA with good quality for molecular genetic research. Therefore, developed screening system can be applicable to find salt stress tolerant black locust genotypes.

Introduction. Soil salinity is an unfavorable natural factor acting in addition to water deficiency in arid areas. Black locust (Robinia pseudoacacia L.) is tree widely used in agroforestry in arid lands. Therefore identification of black locust genotypes exhibiting increased salt stress tolerance is highly important.

Objects. Black locust seeds and seedlings.

Materials and methods. Black locust seeds were germinated without or with NaCl in irrigation water (range of concentrations 0-300 mM). We measured seed germination rate, weight and length of 46 days old seedlings and some stress related biochemical markers.

Results and conclusions. We found black locust seed germination capacity decreased with an increase in the concentration of NaCl (from 93% at 0 mM to 9% at 300 mM). Seedlings grown in the presence of NaCl had some morphological alterations and changes in the biochemical marker values. Strong negative correlation between NaCl concentration in irrigation water and aerial part length to root length ratio was observed. Proposed DNA extraction method allowed to prepare DNA with good quality for molecular genetic research. Developed screening system could be used to find salt stress tolerant black locust seedling for following identification of stress tolerant genotypes.

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