ABA-deficient Mutant Az34 Research Articles

МОРФОГЕНЕЗ IN VITRO В КАЛЛУСАХ ЯЧМЕНЯ: ВЛИЯНИЕ НАФТИЛФТАЛАМОВОЙ КИСЛОТЫ

Morphogenesis in vitro in barley calli of the cv. Steptoe and its ABA-deficient mutant AZ34 under the influence of an inhibitor of polar auxin transport, N-1-naphthylphthalamic acid (NPA), was studied by light microscopy The formation of «polymeric» embryoids (polyembryoids), characterized by the presence of multiple, often fused (fasciated) organs in varying degrees, in calli of cv. Steptoe and the formation of fasciated roots in calli of AZ34 were revealed. The distribution of IAA in calli was studied by the immunolocalization method. It was found that the most intense staining for IAA in both genotypes was found in fasciated roots. The important role of the polar transport of auxins in the regulation of plant morphogenesis in vitro has been confirmed.

Abscisic acid mediates barley rhizosheath formation under mild soil drying by promoting root hair growth and auxin response.

Soil drying enhances root ABA accumulation and rhizosheath formation, but whether ABA mediates rhizosheath formation is unclear. Here, we used the ABA-deficient mutant Az34 to investigate molecular and morphological changes by which ABA could affect rhizosheath formation. Mild soil drying with intermittent watering increased rhizosheath formation by promoting root and root hair elongation. Attenuated root ABA accumulation in Az34 barley constrained the promotion of root length and root hair length by drying soil, such that Az34 had a smaller rhizosheath. Pharmacological experiments of adding fluridone (an ABA biosynthesis inhibitor) and ABA to drying soil restricted and enhanced rhizosheath formation respectively in Az34 and wild-type Steptoe barley. RNA sequencing suggested that ABA accumulation mediates auxin synthesis and responses and root and root hair elongation in drying soil. In addition, adding indole-3-acetic acid (IAA) to drying soil increased rhizosheath formation by promoting root and root hair elongation in Steptoe and Az34 barley. Together, these results show that ABA accumulation induced by mild soil drying enhance barley rhizosheath formation, which may be achieved through promoting auxin response.

ВЛИЯНИЕ ФЛУРИДОНА НА СОДЕРЖАНИЕ ГОРМОНОВ В КАЛЛУСАХ ЯЧМЕНЯ СОРТА STEPTOE И ЕГО АБК‑ДЕФИЦИТНОГО МУТАНТА AZ34

The effect of the inhibitor of endogenous ABA synthesis fluridone on the content and distribution of endogenous ABA and IAA in the calli of ABA-deficient mutant AZ34 barley and its parental cultivar Steptoe was studied using the methods of immuno-enzymatic solid-phase assay and immunolocalization of phytohormones. It was found that by the 4th week of in vitro culture, fluridone causes a significant decrease in the ABA level in calli of both genotypes compared to the control, and the inhibitory effect of fluridone in AZ34 is more pronounced than in Steptoe. In the calli of both genotypes, a significant increase in the IAA content was revealed against the background of a decrease in the ABA content upon treatment with fluridone as compared to the control. It was concluded that ABA plays an important role in the process of embryoido-genesis in vitro.

Dynamics of the contents and distribution of ABA, auxins and aquaporins in developing caryopses of an ABA-deficient barley mutant and its parental cultivar

AbstractDynamics of abscisic acid (ABA) and indole-3-acetic acid (IAA) contents were followed in developing barley caryopses of the ABA-deficient mutant AZ34 and its parental cultivar Steptoe. Distribution of these hormones and HvPIP2 aquaporins (AQPs) was studied with the help of immunohistochemical methods in the roots and coleorhiza of developing embryos. In Steptoe, maturation of the caryopsis was accompanied by vast accumulation of ABA, while this hormone accumulated more slowly in the caryopsis of AZ34 and its content was lower than in Steptoe. Accumulation of ABA was accompanied by a decline in IAA level in the developing caryopsis, the process being delayed in AZ34 in accordance with the slower accumulation of ABA. ABA accumulated to high levels in the coleorhiza cells of Steptoe, while the effect was absent in AZ34. The high level of ABA was likely to be important for maintaining the barrier function of the coleorhiza, preventing germination of seminal roots and enabling seed dormancy, while the absence of ABA accumulation in coleorhiza of AZ34 may be responsible for the initiation of root germination inside the caryopsis. The abundance of HvPIP2 AQPs in the seminal roots was higher at the beginning of maturation of Steptoe caryopsis and declined afterwards, while the levels of APQs increased later in AZ34 in accordance with the delay in ABA accumulation. These results suggest the importance of ABA accumulation in coleorhiza for preventing precocious growth of seminal roots, and suggest regulation of IAA and aquaporin levels by this hormone during maturation of embryos.

Somatic Embryogenesis in Wheat and Barley Calli in vitro Is Determined by the Level of Indoleacetic and Abscisic Acids

The effect of a number of phytohormones (IAA, ABA, and cytokinines) on the induction of in vitro somatic embryogenesis in callus cultures of wheat, cultivar Bashkirskaya 26, barley, cultivar Steptoe, and its ABA-deficient mutant AZ34 has been studied. It was shown that the ability or inability for somatic embryogenesis in callus tissue of both wheat and barley is determined by the IAA : ABA ratio. However, the level of cytokinines was similar in both embryogenic and nonembryogenic calli of each object studied. The analysis of callus tissue of the ABA-deficient mutant AZ34 revealed the stimulating role of ABA not only in the induction of somatic embryogenesis but also in the formation of normal embryos.

Melatonin enhances cold tolerance in drought-primed wild-type and abscisic acid-deficient mutant barley.

Melatonin is involved in multiple plant developmental processes and various stress responses. To explore the roles of melatonin played as well as its association with abscisic acid (ABA) in a process of drought priming-induced cold tolerance (DPICT), a wild-type barley and its ABA-deficient mutant Az34 counterpart were selected for comparison, in which the effects of melatonin application (either foliarly or rhizospherically) and/or drought priming on the cold tolerance of both types of barleys were systematically investigated. It was demonstrated that the early drought priming induced an increase of endogenous melatonin production, which is not ABA dependent. In addition, exogenously applied melatonin resulted in higher ABA concentration in the drought-primed plants than in the nonprimed plants when exposed to cold stress, indicating that ABA responded in a drought-dependent manner. The interplay of melatonin and ABA leads to plants maintaining better water status. Drought priming-induced melatonin accumulation enhanced the antioxidant capacity in both chloroplasts and mitochondria, which sustained the photosynthetic electron transport in photosynthetic apparatus of the plants under cold stress. These results suggest that the exogenous melatonin application enhances the DPICT by modulating subcellular antioxidant systems and ABA levels in barley.

Jasmonate-induced gene expression of barley (Hordeum vulgare) leaves -- the link between jasmonate and abscisic acid

In barley leaves a group of genes is expressed in response to treatment with jasmonates and abscisic acid (ABA) [21]. One of these genes coding for a jasmonate-induced protein of 23 kDa (JIP-23) was analyzed to find out the link between ABA and jasmonates by recording its expression upon modulating independently, the endogenous level of both of them. By use of inhibitors of JA synthesis and ABA degradation, and the ABA-deficient mutant Az34, as well as of cultivar-specific differences, it was shown that endogenous jasmonate increases are necessary and sufficient for expression of this gene. The endogenous rise of ABA did not induce synthesis of JIP-23, whereas exogenous ABA did not act via jasmonates. Different signalling pathways are suggested and discussed.

PH-regulated leaf cell expansion in droughted plants is abscisic acid dependent

Elongation rates of barley (Hordeum vulgare L. cv Hanna) leaves decreased with decreasing soil water content, whereas the pH of xylem sap increased from 5.9 to 6.9 over 6 d as the soil dried. The reduction in leaf-elongation rate (LER) was correlated with the increase in sap pH. Artificial sap buffered to different pH values was fed via the subcrown internode to derooted seedlings. Although leaves elongated at in planta rates when fed artificial sap at a well-watered pH of 6.0, LER declined with increasing sap pH. This effect persisted in the light and in the dark. pH had no effect on the relative water content or the bulk abscisic acid (ABA) concentration of the growing zone of these leaves. LERs of the ABA-deficient mutant Az34 were uniformly high over the pH range tested, whereas those of its isogenic wild-type cultivar Steptoe were reduced as the artificial sap pH was increased from 6.0 to 7.0. However, supplying a well-watered concentration of ABA (3 x 10(-8) M) in the artificial xylem sap restored the pH response of the Az34 mutant. The results suggest that increased xylem sap pH acts as a drought signal to reduce LER via an ABA-dependent mechanism.