Discussion Abiotic Stress Tolerance and Corresponding Strategies for Maize Breeding in Southwest Region of China

Maize is an important food crop, as well as the irreplaceable feed and industrial materials, having huge market demand in China. Southwestern region of China is the third largest main maize producing zone, and the frequent occurrence of abiotic stress conditions such as drought, heat, cold, wet shaded stress have severely affected the development of maize production, causing low and unstable corn yields, severely restricting the maize industry development in the southwest of China. This paper preliminarily describes the maize resistance to abiotic stresses in southwestem region of China, putting forward the countermeasures and the key research direction in the practice of breeding in order to provide reference for the cultivation of new varieties with high yield and stress resistance, and improving the levels of maize stress resistance breeding in southwestern region of China.

Soybean GmMYB76, GmMYB92, and GmMYB177 genes confer stress tolerance in transgenic Arabidopsis plants

MYB-type transcription factors contain the conserved MYB DNA-binding domain of approximately 50 amino acids and are involved in the regulation of many aspects of plant growth, development, metabolism and stress responses. From soybean plants, we identified 156 GmMYB genes using our previously obtained 206 MYB unigenes, and 48 were found to have full-length open-reading frames. Expressions of all these identified genes were examined, and we found that expressions of 43 genes were changed upon treatment with ABA, salt, drought and/or cold stress. Three GmMYB genes, GmMYB76, GmMYB92 and GmMYB177, were chosen for further analysis. Using the yeast assay system, GmMYB76 and GmMYB92 were found to have transactivation activity and can form homodimers. GmMYB177 did not appear to have transactivation activity but can form heterodimers with GmMYB76. Yeast onehybrid assay revealed that all the three GmMYBs could bind to cis-elements TAT AAC GGT TTT TT and CCG GAA AAA AGG AT, but with different affinity, and GmMYB92 could also bind to TCT CAC CTA CC. The transgenic Arabidopsis plants overexpressing GmMYB 76 or GmMYB177 showed better performance than the GmMYB92-transgenic plants in salt and freezing tolerance. However, these transgenic plants exhibited reduced sensitivity to ABA treatment at germination stage in comparison with the wild-type plants. The three GmMYB genes differentially affected a subset of stress-responsive genes in addition to their regulation of a common subset of stress-responsive genes. These resuits indicate that the three GmMYB genes may play differential roles in stress tolerance, possibly through regulation of stress-responsive genes.

Mapping of QTLs for Leaf Malondialdehyde Content Associated with Stress Tolerance in Rice

Malondialdehyde （MDA） is the final product of lipid peroxidation, and MDA content can reflect the stress tolerance of plants. To map QTLs conditioning the MDA content in rice leaves, a recombinant inbred line （RIL） population with 247 lines derived from an indica-indica cross Zhenshan 97BxMilyang 46, and a linkage map consisting of 207 DNA markers were used. The RIL population showed a transgressive segregation in the MDA content of rice leaves. Two QTLs for the MDA content in rice leaves were detected in the intervals RG532-RG811 and RG381-RG236 on chromosome 1, with the additive effects from maternal and paternal parents, accounting for 4.33% and 4.62% of phenotype variations, respectively.

Production of Transgenic Tall Fescue Plants with Enhanced Stress Tolerances by Agrobacterium tumefaciens-Mediated Transformation

In order to improve stress tolerances of turf-type tall fescue （Festuca arundinacea Schreb.）, Agrobacterium tumefaciens strain EHA105 carrying plasmid pCMD containing stress tolerance-related CBF1 gene from Arabidopsis thaliana was used to transform mature seeds-derived embryogenic calli of four cultivars. A total of 112 transgenic plants were regenerated from 32 independent lines and verified by histochemical detection of GUS activity, PCR assay and Southern hybridization analysis. The transformation frequency ranged from 0.92 to 2.87% with apparent differences among the cultivars. Stress tolerances of transgenic plants were enhanced, which was shown by the facts that transgenic plants had distinct growth superiority and significantly higher survival rate than non-transformed ones under high salinity and high osmosis stresses, and that relative electronic conductivity of in vitro leaves treated with low and high temoeratures, dehvdration and high salinity stresses was 25-30% lower in transgenic plants than in control plants.In addition,it was observed that growth of transgenic plants was inhibited due to constitutive overexpression of CBF1 gene under normal environmental conditions.

Roles of MicroRNAs in Plant Stress Tolerance

MicroRNAs （miRNAs） are one kind of small RNA in all eukaryote. MicroRNAs can regulate gene expression of eukaryote; they widely participate in every physiological process. They can block mRNA expression or cleave mRNA by complement to target mRNA. Scholars estimate miRNA genes occuping about 1% of genome, but they can regulate 10%-30% genes of whole genome. The genes are regulated by miRNA including signal proteins, enzymes, transcription factors, and so on. In the field of plant research, the start of miRNA research is later, but it is proved that plant miRNAs are important to every plant physiological process. Now miRNA has become the hotspot of plant molecular biology research. This paper introduced the biology function, action mechanism, researching method and recently development of microRNAs, also focused on advances in plant microRNAs. This paper has important reference value for plant stress tolerance miRNA research.

Stress-Activated Protein Kinase OsSAPK7 Regulates Salt- Stress Tolerance by Modulating Diverse Stress-Defensive Responses in Rice

Soil salinity is an environmental threat limiting rice productivity.Identification of salinity tolerance genes and exploitation of their mechanisms in plants are vital for crop breeding.In this study,the function of stress-activated protein kinase 7(OsSAPK7),a SnRK2 family member,was characterized in response to salt stress in rice.Compared with variety 9804,OsSAPK7-overexpression plants had a greater survival rate,increased chlorophyll and proline contents,and superoxide dismutase and catalase activities at the seedling stage under salt-stress conditio ns,as well as decreased sodium potassium ratio(Na+/K+)and malondialdehyde contents.After salt stress,the OsSAPK7 knockout plants had lower survival rates,in creased Na^K*ratios and malomdiadehyde contents,and decreased physiological parameters compared with 9804.These changes in transgenic lines suggested that OsSAPK7 increased the salt tolerance of rice by modulating ion homeostasis,redox reactions and photosynthesis.The results of RNA-Seq indicated that genes involved in redox-dependent signaling pathway,photosynthesis and zeatin synthesis pathways were significantly down-regulated in the OsSAPK7 knockout line compared with 9804 un der salt-stress conditio n,which con firmed that OsSAPK7 positively regulated salt tolera nee by modulating diverse stress-defe nsive resp on ses in rice.These findings provided novel in sights for the genetic improvement of rice and for understanding the regulatory mechanisms of salt-stress toleranee.

Phenotypic, Stress Tolerance and Plant Growth Promoting Characteristics of Rhizobial Isolates from Selected Wild Legumes of Semiarid Region, Tirupati, India

Rhizobia are vital for nitrogen input, fertility of soil and legume plant growth. Knowledge on rhizobial diversity from arid and semiarid areas is important for dry land agriculture in the context of climatic change and for economic utilization. This study provides morphological, biochemical, stress tolerance and plant growth promoting characteristics of fifteen rhizobial isolates from the nodules of same number of wild legumes and one isolate from cultivated Arachis hypogea from semi-arid region, Tirupati. The bacterial isolates were confirmed as rhizobia based on colony morphology and biochemical tests. Based on the colour change of YMA-BTB medium, eight isolates were identified as slow growers and six were fast growers. The isolates differed in growth pattern, colony morphology, antibiotic resistance at higher concentrations and uniformity in utilization of carbon and nitrogen sources. The isolates are tolerant to NaCl up to one percent, displayed normal growth at temperatures 28℃ - 30℃, at neutral pH and poor growth at pH 5and 9. The isolates varied in the production of EPS and IAA, positive for phosphate solubilization and siderophore formation. This functional diversity displayed by the isolates can be utilised for the legume crop production by cross inoculation.

G-protein β subunit AGB1 positively regulates salt stress tolerance in Arabidopsis

The heterotrimeric GTP-binding proteins（G-proteins） in eukaryotes consisted of α, β and γ subunits and are important in molecular signaling by interacting with G-protein-coupled receptors（GPCR）, on which to transduce signaling into the cytoplast through appropriate downstream effectors. However, downstream effectors regulated by the G-proteins in plants are currently not well defined. In this study, the transcripts of AGB1, a G protein β subunit gene in Arabidopsis were found to be down-regulated by cold and heat, but up-regulated by high salt stress treatment. AGB1 mutant（agb1-2） was more sensitive to high salt stress than wild-type（WT）. Compared with WT, the cotyledon greening rates, fresh weight, root length, seedling germination rates and survival rates decreased more rapidly in agb1-2 along with increasing concentrations of Na Cl in normal（MS） medium. Physiological characteristic analysis showed that compared to WT, the contents of chlorophyll, relative proline accumulation and peroxidase（POD） were reduced, whereas the malonaldehyde（MDA） content and concentration ratio of Na＋/K＋ were increased in agb1-2 under salt stress condition. Further studies on the expression of several stress inducible genes associated with above physiological processes were investigated, and the results revealed that the expressions of genes related to proline biosynthesis, oxidative stress response, Na＋ homeostasis, stress- and ABAresponses were lower in agb1-2 than in WT, suggesting that those genes are possible downstream genes of AGB1 and that their changed expression plays an important role in determining phenotypic and physiologic traits in agb1-2. Taken together, these findings indicate that AGB1 positively regulates salt tolerance in Arabidopsis through its modulation of genes transcription related to proline biosynthesis, oxidative stress, ion homeostasis, stress- and ABA-responses.

Research Progress of miR164 and Target Gene NAC in Regulating Sugarcane Stress Tolerance

Studies have shown that miR164 is a highly conserved miRNA family between monocot and dicotyledonous plants,and it plays an important role in the growth and development of sugarcane organs and in response to stress.As the main target gene of miR164,NAC transcription factors are mainly regulated at the post-transcriptional level.MiR164:NAC module may play an important role in determining the adaptive response of sugarcane to stress.MiR164 has a regulatory effect on the expression of target gene NAC,and may be closely related to the resistance process of sugarcane to abiotic stress,which provides a reference for using miRNA to carry out sugarcane resistance molecular breeding.

Relationship between Ethanol and 2-phenylethanol Stress Tolerance and Fatty Acid Compositions of Saccharomyces cerevisiae

In the present study, we investigated of the ethanol （Eth）, 2-phenylethanol （2-PE） and ethanol/2-phenylethanol （Eth/2-PE） stress tolerance and established relationship between stress tolerance and fatty acid compositions of wine yeast strain S. cerevisiae UCM Y-524 in relation to different cultivation factors. Changes in cell membrane fatty acid profiles studied under different temperature and media. The active oxygenation, semi-aerobic cultivation supplemented Tween 80 conditions were used in different combinations. The concentrations of 2-PE and Eth, the fatty acids methyl esters （FAMES） in the media were measured by GC-MS analyses. 2-PE, Eth, FAMES were identified by NIST02 MS database at the 2-PE and Eth standard solution （Merck, Germany）, bacterial FAMES standard （Supelco）. Palmitoleic and oleic acid were dominated for S. cerevisiae UCM Y-524. The unsaturated/saturated fatty acid （UFA/SFA） ratio was higher than five for S. cerevisiae UCM Y-524 at 14 ~C in simple medium and about more than three for other conditions. It has been found that yeast cells grown in the presence of the 2-PE, Eth and Eth/2-PE appear to increase the amount of unsaturated fatty acids, especially oleic acid, in cellular lipids especially under active oxygenation supplemented with Tween 80. S. cerevisiae UCM Y-524 has a higher tolerance under 2-PE, Eth and Eth/2-PE stress at 28 ~C in a simple medium under aerobic conditions. A higher concentration of unsaturated fatty acids in the membrane S. cerevisiae UCM Y-524 and a lower temperature under aerobic conditions, has been considered to have a positive influence on the excretion of 2-phenylethanol.

IiWRKY34 positively regulates yield, lignan biosynthesis and stress tolerance in Isatis indigotica

Yield potential,pharmaceutical compounds production and stress tolerance capacity are 3 classes of traits that determine the quality of medicinal plants.The autotetraploid Isatis indigotica has greater yield,higher bioactive lignan accumulation and enhanced stress tolerance compared with its diploid progenitor.Here we show that the transcription factor IiWRKY34,with higher expression levels in tetraploid than in diploid I.indigotica,has large pleiotropic effects on an array of traits,including biomass growth rates,lignan biosynthesis,as well as salt and drought stress tolerance.Integrated analysis of transcriptome and metabolome profiling demonstrated that IiWRKY34 expression had far-reaching consequences on both primary and secondary metabolism,reprograming carbon flux towards phenylpropanoids,such as lignans and flavonoids.Transcript-metabolite correlation analysis was applied to construct the regulatory network of IiWRKY34 for lignan biosynthesis.One candidate target Ii4CL3,a key rate-limiting enzyme of lignan biosynthesis as indicated in our previous study,has been demonstrated to indeed be activated by IiWRKY34.Collectively,the results indicate that the differentially expressed IiWRKY34 has contributed significantly to the polyploidy vigor of I.indigotica,and manipulation of this gene will facilitate comprehensive improvements of I.indigotica herb.

A Bi-Functional Xyloglucan Galactosyltransferase Is an Indispensable Salt Stress Tolerance Determinant in Arabidopsis

Salinity is an abiotic stress that substantially limits crop production worldwide. To identify salt stress tolerance determinants, we screened for Arabidopsis mutants that are hypersensitive to salt stress and designated these mutants as short root in salt medium （rsa）. One of these mutants, rsa3-1, is hypersensitive to NaCI and LiCI but not to CsCI or to general osmotic stress. Reactive oxygen species （ROS） over-accumulate in rsa3-1 plants under salt stress. Gene expression profiling with Affymetrix microarray analysis revealed that RSA3 controls expression of many genes including genes encoding proteins for ROS detoxification under salt stress. Map-based cloning showed that RSA3 encodes a xyloglucan galactosyltransferase, which is allelic to a gene previously named MUR3/KAM1. The RSA3/ MUR3/KAMl-encoded xylogluscan galactosyltransferase regulates actin microfilament organization （and thereby con- tributes to endomembrane distribution） and is also involved in cell wall biosynthesis. In rsa3-1, actin cannot assemble and form bundles as it does in the wild-type but instead aggregates in the cytoplasm. Furthermore, addition of phal- Ioidin, which prevents actin depolymerization, can rescue salt hypersensitivity of rsa3-1. Together, these results sug- gest that RSA3/MUR3/KAM1 along with other cell wall-associated proteins plays a critical role in salt stress tolerance by maintaining the proper organization of actin microfilaments in order to minimize damage caused by excessive ROS.

The role of tocopherol cyclase in salt stress tolerance of rice(Oryza sativa)

Tocopherols synthesized exclusively by photosynthetic organisms are major antioxidants in biomembranes.In plants,tocopherol cyclase(TC/VTE1) catalyzes the conversion of 2,3-dimethyl-5-phytyl-1,4-benzoquinone(DMPBQ) to γ-tocopherol.In the present study,OsVTE1,which encodes a rice tocopherol cyclase ortholog,was cloned and characterized.OsVTE1 was induced significantly by abiotic stresses such as high salt,H2O2,drought,cold and by the plant hormones ABA and salicylic acid.The tissue-specific expression pattern and OsVTE1-promoter GUS activity assay showed that OsVTE1 was mainly expressed in the leaf,and also could be detected in the root,stem and panicle.Compared with control plants,transgenic plants with OsVTE1 RNA interference(OsVTE1-RNAi) were more sensitive to salt stress whereas,in contrast,transgenic plants overexpressing OsVTE1(OsVTE1-OX) showed higher tolerance to salt stress.The DAB in vivo staining showed that OsVTE1-OX plants accumulated less H2O2 than did control plants.

Expression of Acyl-lipid 12-desaturase Gene in Prokaryotic and Eukaryotic Cells and Its Effect on Cold Stress Tolerance of Potato

We report the expression profile of acyl-lipid △12-desaturase （desA） gene from Synechocystis sp. PCC6803 and its effect on cell membrane lipid composition and cold tolerance in prokaryotic （Escherichia coil） and eukaryotic （Solanum tuberosum） cells. For this purpose, a hybrid of desA and reporter gene encoding thermostable lichenase （licBM3） was constructed and used to transform these cells. The expression of this hybrid gene was measured using qualitative （Petri dish test, electrophoregram and zymogram） and quantitative methods （spectrometry and gas liquid chromatography assays）. The maximum level of linoleic acid in the bacterial cells containing hybrid gene was 1.9% of total fatty acids. Cold stress tolerance assays using plant damage index and growth parameters showed that cold tolerance was enhanced in primary transgenic lines because of increased unsaturated fatty acid concentration in their lipids. The greatest content of 18：2 and 18：3 fatty acids in primary transgenic plants was observed for lines 2 （73%） and 3 （41%）. Finally, our results showed that desaturase could enhance tolerance to cold stress in potato, and desaturase and lichenase retain their functionality in the structure of the hybrid protein where the enzymatic activity of target gene product was higher than in the case of reporter lichenase gene absence in the construction.

Overexpression of AmDUF1517 enhanced tolerance to salinity, drought, and cold stress in transgenic cotton

As abiotic stresses become more severe as a result of global climate changes, the growth and development of plants are restricted. In the development of agricultural crops with greater stress tolerance, AmDUF1517 had been isolated from the highly stress-tolerant shrub Ammopiptanthus mongolicus, and can significantly enhance stress tolerance when inserted in Arabidopsis thaliana. In this study, we inserted this gene into cotton to analyze its potential for conferring stress tolerance. Two independent transgenic cotton lines were used. Southern blot analyses indicated that AmDUF1517 was integrated into the cotton genome. Physiological analysis demonstrated that AmD UF1517-transgenic cotton had stronger resistance than the control when treated with salt, drought, and cold stresses. Further analysis showed that trans-AmD UF1517 cotton displayed significantly higher antioxidant enzyme（superoxide dismutase（SOD）, peroxidase（POD）, catalase（CAT）, and glutathione S-transferase（GST）） activity and less reactive oxygen species（ROS） accumulation, which suggests that overexpression of AmDUF1517 can improve cotton resistance to stress by maintaining ROS homeostasis, as well as by alleviating cell membrane injury. These results imply that AmDUF1517 is a candidate gene in improving cotton resistance to abiotic stress.

Selection of Maize Genotypes with Tolerance to Osmotic Stress Associated with Salinity

Thirteen different inbred lines in relation to the type of grain and life cycles were characterized by testing for osmotic stress associated with salinity. The identification of tolerant genotypes would be an effective strategy to overcome the saline stress. Osmotic stress reduces immediately the expansion of the roots and young leaves which determine a reduction in the size of the plant. A completely randomized design was adopted to test seedlings under controlled conditions of light and temperature. Two treatments were used: 0 mM NaCl (as control) and 100 mM NaCl. After 15 days of complete salinization, the seedlings were harvested and several morphological traits were measured. The morphological traits of growth were leaf growth (Ar1, Ar2, Ar3 and Ar4), dry masses of shoot and root (SDM and RDM, respectively). Also, traits associated with water economy were registered: leaf water loss (LWL) and relative water content (RWC). The morphological traits were expressed in relative terms, while the traits associated with the economy of water were expressed in absolute terms. Uni and multivariate techniques were applied to identify genotypes with divergent behaviors to osmotic stress tolerance. Also, a Tolerance Index was employed to identify superior genotypes. Four clusters were obtained after applying a Cluster Analysis and Principal Component Analysis (PCA). The genotypes were compared to each other with a test of DMS. The results obtained with different statistical techniques converged. Some variables presented a differential weight classification of genotypes. The morphological traits like RDM, SDM, Ar3, Ar4 and Ar5 were the most discriminating. Tolerance Index allowed to classify genotypes, thus SC2 and AD3 lines were that reached highest value of the index and therefore would be tolerant lines, while AF3 and LP3 had a low index and were seen as sensible.

Reduction of methylviologen-mediated oxidative stress tolerance in antisense transgenic tobacco seedlings through restricted expression of StAPX

Ascorbate peroxidases are directly involved in reactive oxygen species (ROS) scavenging by reducing hydrogen peroxide to water. The tomato thylakoid-bound ascorbate peroxidase gene (StAPX) was introduced into tobacco. RNA gel blot analysis confirmed that StAPX in tomato leaves was induced by methylviologen-mediated oxidative stress. The sense transgenic seedlings exhibited higher tAPX activity than that of the wild type (WT) plants under oxidative stress conditions, while the antisense seedlings exhibited lower tAPX activity. Lower APX activities of antisense transgenic seedlings caused higher malondialdehyde contents and relative electrical conductivity. The sense transgenic seedlings with higher tAPX activity maintained higher chlorophyll content and showed the importance of tAPX in maintaining the optimal chloroplast development under methylviologen stress conditions, whereas the antisense lines maintained lower chlorophyll content than WT seedlings. Results indicated that the over-expression of StAPX enhanced tolerance to methylviologen-mediated oxidative stress in sense transgenic to- bacco early seedlings, whereas the suppression of StAPX in antisense transgenic seedlings showed high sensitivity to oxidative stress.

Nucleus-Encoded Thylakoid Protein,OsY3IP1,Confers Enhanced Tolerance to Saline and Alkaline Stresses in Rice

Abiotic stress confers serious damage to the photosynthetic machinery,often resulting in plant growth inhibition.Hypothetical chloroplast open reading frame 3(Ycf3)-interacting protein 1(Y3IP1)is a nucleus-encoded thylakoid protein and plays an essential role in the assembly of photosystem I.The full-length cDNA over-expresser(FOX)gene-hunting system is an approach using systemically generated gain-of-function mutants.Among the FOX-rice lines,a line CE175 overexpressing rice Y3IP1gene(Os Y3IP1)displayed less inhibition of root growth under saline(NaCl)stress.The expression of Os Y3IP1 was up-regulated under saline and alkaline(Na2CO3)stresses in the rice variety Kitaake.After saline and alkaline treatments,transgenic Kitaake overexpressing OsY3IP1-GFP(OsY3IP1-GFPox/Kit)displayed higher levels of chlorophyll content compared to Kitaake.Under the stress conditions,the maximum quantum yield of photosystem II photochemistry levels was higher in OsY3IP1-GFPox/Kit than in Kitaake.The increased tolerance conferred by OsY3IP1 overexpression correlated with reduced reactive oxygen species accumulation.Our data provide new insights into the possible role of OsY3IP1 in the pathway suppressing photooxidative damage under stress conditions.These features can be further exploited to improve saline and alkaline tolerances of rice plants in future.

Ethanol Fermentation by High-Stress-Tolerance Aquatic Yeasts and Their Mutants

Bioethanol is thought to be a renewable source of energy, because the biomasses used to make ethanol, such as sugar cane and its residual substance, molasses, are resources that can be continuously produced. But the practical use of ethanol to replace fossil fuels or atomic energy has been limited, because the production efficiencies of ethanol in relation to its substrates are not so high. Thus, for industrial production of the bioethanol, yeast fermentation would ideally be carried out in biomasses containing more highly concentrated carbohydrates. However, the environmental stresses in highly concentrated cultures might weaken the yeast’s physiological activities. From various kinds of aquatic yeast with stress tolerance, Torulaspora derbrueckii F2-11 and Wicherhamomyces anomalus AN2-64 were selected as candidates for high-sugar-tolerance yeasts as they showed remarkable growth in the YPD + sorbitol (600 g/L) medium at 25°C for 120 hrs. When the amounts and kinds of sugar alcohols in the cells of the two strains were measured in cultures containing 20 g/L or 400 g/L of D-glucose, maltose, or sucrose, the main two sugar alcohols that accumulated as the sugar concentration increased were glycerol and arabitol. Mutation by ethyl methanesulfonate of the parent strains T. derbrueckii F2-11 and W. anomalus AN2-64 induced mutants F2-11M or AN2-64M, which showed higher sugar, heat, and ethanol tolerances than their respective parents. Ethanol productivities and sugar assimilation activities of the mutants were also higher than those of the parents in the 25% (v/v) molasses.

JAZ proteins:Key regulators of plant growth and stress response

The jasmonate ZIM-domain(JAZ)family of proteins serves as co-receptors and transcriptional repressors of jasmonic acid(JA)in plants.Their functional diversity and multiple roles make them important components of the regulatory network of JA and other hormonal signaling pathways.In this review,we provide an overview of the latest findings on JAZ family proteins and emphasize their roles in plant growth and development,and response to biotic and abiotic stress,along with their underlying mechanisms.Moreover,existing challenges and future applications are outlined with the aim of offering a reference for further research on JAZ proteins in the context of plant physiology.