Transgenic barley with overexpressed PTrx increases aluminum resistance in roots during germination.

A transgenic barley line (LSY-11-1-1) with overexpressed Phalaris coerulescens thioredoxin gene (PTrx) was employed to measure the growth, protein oxidation, cell viability, and antioxidase activity in barley roots during germination on the presence of 2 mmol/L AlCl(3) on filter paper. The results show that (1) compared with the non-transgenic barley, LSY-11-1-1 had enhanced root growth, although both were seriously inhibited after AlCl(3) treatment; (2) the degree of protein oxidation and loss of cell viability in roots of LSY-11-1-1 were much less than those in roots of non-transgenic barley, as reflected by lower contents of protein carbonyl and Evans blue uptakes in LSY-11-1-1; (3) activities of catalase (CAT), glutathione peroxidase (GPX), ascorbate peroxidase (APX), and glutathione reductase (GR) in LSY-11-1-1 root tips were generally higher than those in non-transgenic barley root tips, although these antioxidase activities gave a rise to different degrees in both LSY-11-1-1 and non-transgenic barley under aluminum stress. These results indicate that overexpressing PTrx could efficiently protect barley roots from oxidative injury by increasing antioxidase activity, thereby quenching ROS caused by AlCl(3) during germination. These properties raise the possibility that transgenic barley with overexpressed PTrx may be used to reduce the aluminum toxicity in acid soils.

The level of expression of thioredoxin is linked to fundamental properties and applications of wheat seeds.

Work with cereals (barley and wheat) and a legume (Medicago truncatula) has established thioredoxin h (Trx h) as a central regulatory protein of seeds. Trx h acts by reducing disulfide (S-S) groups of diverse seed proteins (storage proteins, enzymes, and enzyme inhibitors), thereby facilitating germination. Early in vitro protein studies were complemented with experiments in which barley seeds with Trx h overexpressed in the endosperm showed accelerated germination and early or enhanced expression of associated enzymes (alpha-amylase and pullulanase). The current study extends the transgenic work to wheat. Two approaches were followed to alter the expression of Trx h genes in the endosperm: (1) a hordein promoter and its protein body targeting sequence led to overexpression of Trx h5, and (2) an antisense construct of Trx h9 resulted in cytosolic underexpression of that gene (Arabidopsis designation). Underexpression of Trx h9 led to effects opposite to those observed for overexpression Trx h5 in barley-retardation of germination and delayed or reduced expression of associated enzymes. Similar enzyme changes were observed in developing seeds. The wheat lines with underexpressed Trx showed delayed preharvest sprouting when grown in the greenhouse or field without a decrease in final yield. Wheat with overexpressed Trx h5 showed changes commensurate with earlier in vitro work: increased solubility of disulfide proteins and lower allergenicity of the gliadin fraction. The results are further evidence that the level of Trx h in cereal endosperm determines fundamental properties as well as potential applications of the seed.

Protective role of exogenous nitric oxide against oxidative-stress induced by salt stress in barley (Hordeum vulgare).

To probe into the potential of relieving the oxidative damage of salt stress, we investigated the protective role of nitric oxide on barley under salt stress. Salt stress resulted in increased ion leakage, lipid peroxidation and protein oxidation in barley leaves. Simultaneous treatments of barley leaves with 50 microM sodium nitroprusside, a nitric oxide donor, alleviated the damage of salt stress, reflected by decreased ion leakage, and malendialdehyde (MDA), carbonyl, and hydrogen peroxide content in barley leaves. The presence of the nitric oxide donor increased the activities of superoxide dismutases (SOD), ascorbate peroxidases (APX), and catalases (CAT). Meantime, sodium nitroprusside addition increased accumulation of ferritin at the protein level, indicating that nitric oxide directly regulated ferritin accumulation. These results suggested that nitric oxide can effectively protect seedlings from salt stress damage by enhancing activities of antioxidant enzymes to quench the excessive reactive oxygen species caused by salt stress and inducing the increase of ferritin accumulation to chelate larger number of ferrous ion. Information from this study can be used to improve soil management practices for sustainable use of salt-affected soils in the future.

[Effects of antisense-thioredoxin s gene on expression of endogenous thioredoxin h gene in transgenic wheat seed].

To clarify the function mechanism of antisense-thioredoxin s (anti-trxs) gene in transgenic wheat, the expression pattern of endogenous trxh gene in transgenic line 01TY70-1-17-5 and non-transgenic cultivar 'Yumai 70' were detected by semi-quantitative RT-PCR using wheat actin gene as the endogenous control. The results of analysis of transgenic and non-transgenic seeds in different maturation periods, different tissues and different germinating processes indicated that the mRNA transcript amounts of trxh gene in transgenic line seed were lowered distinctly, though the trxh gene mRNA transcript level varied greatly in different developing and germination stages. The mRNA transcript amounts of trxh gene in transgenic line seed were significantly lower than the control seeds by 20.1% 15-30 d after anthesis. The lowest mRNA transcript amount of trxh gene appeared at 25 day after fluorescence and the difference was significant at the 0.05 level. The analysis of gene expression in different tissues also indicated that the transcript levels of trxh gene in transgenic seed were significantly lower than control seeds in 25 d and 30 d after anthesis. The lowest amounts of mRNA transcript of trxh gene was from the endosperm 25 d after anthesis followed by embryo and then by whole seed. During seed germination, the mRNA transcript amounts of trxh gene in transgenic seed were lower than control seed after imbibing 24 h, but the difference was not significant. The above result demonstrated that foreign antisense trxs gene directly interferes with the expression of the endogenous gene.