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The gene expression chip of a salt-tolerant wheat mutant under salt stress was used to clone a salt-induced gene with unknown functions. This gene was designated as TaSR (Triticum aestivum salt-response gene) and submitted to GenBank under accession number EF580107. Quantitative polymerase chain reaction (PCR) analysis showed that gene expression was induced by salt stress. Arabidopsis and rice (Oryza sativa) plants expressing TaSR presented higher salt tolerance than the controls, whereas AtSR mutant and RNA interference rice plants were more sensitive to salt. Under salt stress, TaSR reduced Na(+) concentration and improved cellular K(+) and Ca(2+) concentrations; this gene was also localized on the cell membrane. β-Glucuronidase (GUS) staining and GUS fluorescence quantitative determination were conducted through fragmentation cloning of the TaSR promoter. Salt stress-responsive elements were detected at 588-1074 bp upstream of the start codon. GUS quantitative tests of the full-length promoter in different tissues indicated that promoter activity was highest in the leaf under salt stress. Bimolecular fluorescence complementation and yeast two-hybrid screening further showed the correlation of TaSR with TaPRK and TaKPP. In vitro phosphorylation of TaSR and TaPRK2697 showed that TaPRK2697 did not phosphorylate TaSR. This study revealed that the novel TaSR may be used to improve plant tolerance to salt stress., (© 2015 Scandinavian Plant Physiology Society.)

Microarray analysis of a salt-tolerant wheat mutant identified a gene of unknown function that was induced by exposure to high levels of salt and subsequently denoted TaSIP (Triticum aestivum salt-induced protein). Quantitative PCR analysis revealed that TaSIP expression was induced not only by salt, but also by drought, abscisic acid (ABA), and other environmental stress factors. Transgenic rice plants that expressed an RNA interference construct specific for a rice gene homologous to TaSIP was more susceptible to salt stress than wild-type rice plants. Subcellular localization studies showed that the TaSIP localized to the cell membrane. Under conditions of salt and drought stress, transgenic Arabidopsis plants that overexpressed TaSIP showed superior physiological properties compared with control plants, including lower Na(+) content and upregulation of several stress resistance genes. Staining of transgenic tissues with β-glucuronidase (GUS) failed to indicate tissue-specific activity of the full-length TaSIP promoter. Quantitative analysis of GUS fluorescence in transgenic plants treated with ABA or salt stress revealed that the region 1,176-1,410 bp from the start codon contained an ABA-responsive element and that the region 579-1,176 bp from the start codon upstream of the exon contained a salt-stress-responsive element. Based on these results, we conclude that the key part of the TaSIP gene is the region of its promoter involved in salt tolerance.

The effects of water-retaining agent (60 kg x hm(-2)) and nitrogen fertilizer (0, 225, and 450 kg x hm(-2)) on the leaf photosynthetic characteristics, chlorophyll content, and water utilization of winter wheat at jointing and grain-filling stages were studied under field conditions. In all treatments, the net photosynthetic rate, stomata conductance, intercellular CO2 concentration, water use efficiency, and chlorophyll content were greater at grain-filling stage than at jointing stage. Under nitrogen fertilization but without water-retaining agent application, the water use efficiency (WUE) of single leaf at jointing stage increased with increasing nitrogen fertilization rate, while the net photosynthetic rate, stomata conductance, intercellular CO2 concentration, and transpiration rate decreased after an initial increase. The chlorophyll content was the highest under 225 kg x hm(-2) nitrogen fertilization. In the treatments of water-retaining agent application, the intercellular CO2 con- centration decreased with increasing nitrogen application rate, but the net photosynthetic rate, transpiration rate, and WUE increased. The application of water-retaining agent or its combination with nitrogen fertilization increased the chlorophyll content, but excessive nitrogen fertilization had lesser effects. At grain-filling stage, applying nitrogen fertilizer alone significantly increased the net photosynthetic rate and WUE, but decreased the stomata conductance, intercellular CO2 concentration, and transpiration rate. The chlorophyll content increased with increasing nitrogen application rate. After applying water-retaining agent and with the increase of nitrogen fertilization rate, the photosynthetic rate and WUE decreased after an initial increase, while the intercellular CO2 concentration and transpiration rate were in adverse but still lower than those without water-retaining agent application. The stomata conductance increased with increasing nitrogen fertilization rate. The chlorophyll content increased significantly under the application of water-retaining agent, but somewhat decreased under the combined application of water-retaining agent and nitrogen fertilizer. The application of both water-retaining agent and nitrogen fertilizer increased the 1000 grain mass, grain yield, and water production efficiency of winter wheat significantly, with the best effect in the treatment of water-retaining agent with 225 kg x hm(-2) nitrogen fertilization.

A field experiment was conducted at the Yuzhou Experimental Base of Henan Province to study the effects of different application rates (0, 30, 60, and 90 kg x hm(-2)) of water-retaining agent (WRA) on the root physiological characteristics, biomass, and grain yield of two winter wheat cultivars Zhengmai-9694 and Aikang-58, aimed to probe into the action mechanisms of WRA on the root system of winter wheat at its different growth stages. The application of WRA decreased the root membrane permeability and soluble sugar content, and increased the root vigor. After the application of WRA, the Zhengmai-9694 at its different growth stages had a greater decrement of root membrane permeability, compared with Aikang-58. In all treatments except 90 kg x hm(-2) of WRA, the root vigor of Aikang-58 was obviously higher than that of Zhengmai-9694. At booting and grain-filling stages, the root soluble sugar content of Zhengmai-9694 decreased much more than that of Aikang-58. In the whole growth period of the two cultivars, their root membrane permeability and soluble sugar content were the lowest in treatment 60 kg x hm(-2) of WRA, and no significant differences were observed between treatments 60 and 90 kg x hm(-2) of WRA. The root vigor of Zhengmai-9694 increased remarkably with the increasing rate of WRA application, while that of Aikang-58 was the highest in treatment 60 kg x hm(-2) of WRA. The application of WRA also increased root biomass, and at jointing and booting stages, the root biomass of Aikang-58 was much higher than that of Zhengmai-9694. However, at grain-filling stage, the biomass of Aikang-58 in treatments 60 and 90 kg x hm(-2) of WRA was lower than that of Zhengmai-9694. Treatment 60 kg x hm(-2) of WRA had the highest grain yield of the two cultivars. It was concluded that WRA had more significant effects on Zhengmai-9694 than on Aikang-58, and applying 60 kg x hm(-2) of WRA could obtain the best effect.

High salt and drought are the main factors affecting agricultural production. Thus, cloning stress-tolerance-related genes and identifying their functions are essential to enhancing crop tolerance to stresses. In this study, a salt-induced unknown wheat (Triticum aestivum L.) gene was identified and cloned according to microarray analysis of salt-tolerant wheat mutant RH8706-49 under salt stress. The gene was named Triticum aestivum salt tolerance-related gene (TaSTRG) and submitted to Genbank (Accession number: EF599631). TaSTRG expression in wheat is induced by multiple stresses including salt, polyethylene glycol (PEG), abscisic acid (ABA), and cold. Transgenic rice plants overexpressing TaSTRG gene showed higher salt and drought tolerance than the control. Under salt stress, the transgenic rice had a lower intracellular Na(+)/K(+) ratio than the control. Under salt and PEG treatments, these TaSTRG overexpressing rice plants had higher survival rate, fresh weight and chlorophyll content, accumulated higher proline and soluble sugar contents, and had significantly higher expression levels of putative proline synthetase and transporter genes than the control plants. These results indicate that the wheat TaSTRG gene could enhance plant tolerance to multiple types of stresses.

The root microsomal proteomes of salt-tolerant and salt-sensitive wheat lines under salt stress were analyzed by two-dimensional electrophoresis and mass spectrum. A wheat V-H(+)-ATPase E subunit protein was obtained whose expression was enhanced by salt stress. In silicon cloning identified the full-length cDNA sequences of nine subunits and partial cDNA sequences of two subunits of wheat V-H(+)-ATPase. The expression profiles of these V-H(+)-ATPase subunits in roots and leaves of both salt-tolerant and salt-sensitive wheat lines under salt and abscisic acid (ABA) stress were analyzed. The results indicate that the coordinated enhancement of the expression of V-H(+)-ATPase subunits under salt and ABA stress is an important factor determining improved salt tolerance in wheat. The expression of these subunits was tissue-specific. Overexpression of the E subunit by transgenic Arabidopsis thaliana was able to enhance seed germination, root growth and adult seedling growth under salt stress.

The Tagsk1 (Triticum asetium L. glycogen synthase kinase 1) gene derived from the genome of wheat salt-tolerance mutant RH8706-49 was cloned by PCR. The special primers designed according to full length cDNA sequence of Tagsk1 (AF525086). A binary expression vector pBI121-gsk1 containing Gus and Tagsk1 was constructed. And pBI121-gsk1 was introduced into the callus induced from mature embryos of salt-sensitive wheat H8706-34 and cv. China Spring by particle bombardment. The transformed callus were screened by Kanamycin and 0.5% NaCl. The salt-tolerance callus were obtained, which showed higher ability of salt-tolerance and could diffirentiate roots and buds on the medium containing 0.5% NaCl.

Two dimensional electrophoresis was used to analyse the proteome of the salt-tolerant mutant of wheat (RH8706-49) and the salt-sensitive mutant of wheat (H8706-34) which had been treated by 1% NaCl for 72 hours. After being analysed by MALDI-TOF-MS and Mascot software, the qualitative and quantitative differences were identified between the two materials for five candidate proteins: H+-transporting two-sector ATPase, glutamine synthetase 2 precursor, putative 33 kD oxygen evolving protein of photosystem II and ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit. These five proteins are all belong to chloroplast proteins. They are likely to play a crucial role in keeping the function of the chloroplast and the whole cells when the plant was under salt-stress.

Through the genetic analysis of a F2 population, derived from CMS line 75-3369A (T-type CMS wheat) and the restorer line 7269-10, the result indicated that the restorer line was conditioned by two dominant genes. A F2 population was used to map the fertility restorer (Rf) gene by microsatellite and BSA (bulked segregant analysis). Restorer and sterile DNA pools were established using the extreme fertile and sterile plants of F2 population, respectively. Among the 230 pairs of microsatellite primers, two markers were found polymorphic between the two pools. Linkage analysis showed that microsatellite marker Xgwm136 and Xgwm550 were linked with the two fertility restorer genes, respectively. One of the Rf gene was located on 1AS and the genetic distance between the SSR marker Xgwm136 and this Rf gene was 6.7 cM, the other Rf gene was located on 1BS and with a genetic distance of 5.1 cM to marker Xgwm550.