The Effect of ACC and Salicylic Acid on Germination and GAI1 and LOX2 Genes Expression in Deteriorated Soybean Seeds (Glycine max)

Abstract

Introduction: Soybean ( Glycine Merrill) is the primary source of vegetable oil. Even in desirable conditions, seeds lose their viability in long term storage. Many factors contribute to seed deterioration, including genetic factors, mechanical damage, relative humidity, storage temperature, seed moisture content, existence of microflora, and seed maturity, which reduce seed quality and make seeds unfit for cultivation purposes. Materials and Methods: In order to investigate the effects of seed deterioration on seed germination and also the effects of salicylic acid and ethylene on the improvement of deteriorated seeds of G. max ., accelerated aging test for 0, 6 and 10 days and natural aging test for 6 months were conducted. After aging conditions, seeds were imbibed with 50 µM salicylic acid and 10 µM ACC (precursor of ethylene) for 6 hours at 25 °C. In addition, after natural and accelerated aging tests, a bunch of seeds was used without any hormonal treatment (i.e., dry seeds) as control seeds. The seeds’ germination percentage, total sugar, fructose, and glucose were investigated. Moreover, the gene expression of GAI1 and LOX 1 was measured on dry seeds and under imbibition of water, salicylic acid and ACC at 6, 12 hours using Q-RT-PCR method. Results: The germination results showed that increasing number of aging days led to a decrease in germination. Total sugar content in seeds aged for 6 days did not have a significant difference, as compared with non-aged seeds. However, total sugar content in seeds aged for 10 days was significantly higher than non-aged seeds. Increasing accelerated aging levels from 0 days to 10 days led to increases in glucose and fructose contents in dry seeds. In addition, genes exhibited different expressions in different days and hours. Increasing aging from 0 days to 10 days led to increases in GAI1 gene expression. Moreover, LOX2 expression increased in accelerated aging from 0 to 6 days. LOX2 gene expression in naturally dried aged seeds also increased and was higher than that in non-aged seeds. SA and ACC had different effects on measured values. Conclusion: In general, it can be concluded that the deterioration of seed quality and vigor result from numerous degradation processes and disruption in seeds’ physiological activity. This study showed that aging is associated with an increase in total sugar, glucose and fructose levels. In addition, the expression of the genes involved in the germination is also affected. Increases in LOX2 gene expression were observed in both accelerated aging and natural aging pathways. GAI1 gene expression increased in accelerated aging. However, in normal aging, it decreased.