Overexpression of the peroxidase gene ZmPRX1 increases maize seedling drought tolerance by promoting root development and lignification

Drought is a main abiotic stress factor hindering plant growth,development,and crop productivity.Therefore,it is crucial to understand the mechanisms by which plants cope with drought stress.Here,the function of the maize peroxidase gene ZmPRX1 in drought stress tolerance was investigated by measurement of its expression in response to drought treatment both in a ZmPRX1 overexpression line and a mutant line.The higher root lignin accumulation and seedling survival rate of the overexpression line than that of the wild type or mutant support a role for ZmPRX1 in maize drought tolerance by regulating root development and lignification.Additionally,yeast one-hybrid,Dule luciferase and ChIP-qPCR assays showed that ZmPRX1 is negatively regulated by a nuclear-localized ZmWRKY86 transcription factor.The gene could potentially be used for breeding of drought-tolerant cultivars.

Analysis of Combining Ability of Drought Tolerance in Maize

By using complete-diallel cross design（Griffing method I）, the influence of the combination of different high-yield genotypes of maize on the drought tolerance of their offspring, the general combining ability, the specific combining ability and the back cross effect of drought tolerance between parents and the main genetic parameters for drought tolerance were analyzed. The result indicated that there were significant differences in general combining ability effects（GCA） of maize; there were highly significant differences in special combining ability effects（SCA）; there was no significant difference in reciprocal effects（R）. There were apparent differences in drought tolerance among six parents; to be specific, Zheng 58 had the highest drought tolerance, while PH4CV had the lowest. Improving drought-tolerant parents with Zheng 58, Ji 853 and Xinzi 8717 had gain superiority effects on the increase of drought tolerance in offspring. The influence of the genetic additive effect on the drought tolerance of offsprings varied with different parents and combinations. Therefore, the expression of drought tolerance inheritance genes was determined only by the additive and non-additive genetic effects but had little relationship with reciprocal effects. The selection of drought tolerance of maize should be conducted at higher generations.

Mapping of QTL Associated with Drought Tolerance in a Semi-Automobile Rain Shelter in Maize (Zea mays L.)

Drought is a major constraint in maize production worldwide. We studied quantitative trait loci （QTL） underlying drought tolerance for maize plants grown in two different environments. Traits investigated included ASI, plant height, grain yield, ear height, and ear setting. A genetic linkage map was constructed with 120 simple sequence repeat （SSR） markers based on an F2 population derived from a cross between D5 （resistant parent） and 7924 （susceptible parent）. Correlation and heritability were calculated. QTLs of these traits were identified by composite interval mapping combined with a linkage map covering 1 790.3 cM. The markers were arranged in ten linkage groups. QTL mapping was made of the mean trait performance of the 180 F2：3 population. The results showed five, five, six, four, and five QTLs for ASI, plant height, grain yield, ear height, and ear setting under full irrigation condition, respectively, and four, seven, six, four, and four QTLs for ASI, plant height, grain yield, ear height, and ear setting under severe late stress conditions, respectively. Especially the four QTLs detected for five traits in 2008 and 2009. The universal QTLs information generated in this study will aid in undertaking an integrated breeding strategy for further genetic studies in drought tolerance improvement in maize.

Drought Tolerance Identification and Eevluation of Maize during Flowering Stage in Guangxi

Total 20 maize varieties were subjected to drought stress at flowering stage,and then the relative water content,soluble sugar content,chlorophyll content,malondialdehyde( MDA) content and superoxide dismutase( SOD) activity in their leaves,as well as their yields were determined. The drought tolerance of the physiological and biochemical indexes was scored by five-level scoring method,and the drought tolerance index was calculated by the yield index to comprehensively evaluate the drought tolerance of maize during flowering stage. The results showed that the scores of drought tolerance of the maize varieties ranged from 1. 929 3 to 5. 659 5. Among them,the scores of Zhengda 619,Guidan 162 and Guidan 0810 were greater than 5. 0,followed by Dika 008,Xianyu 30 T60,Xianzhengda 901,Qingnong 68,South America No. 1 and Wanchuan 1306 of which the scores were in the range of 4. 0-5. 0. The drought tolerance indexes were in the range of 0. 410 4-1. 096 3. Among the test maize varieties,the drought tolerance indexes of Guidan 0810,Pacific 99 and Zhengda 619 were greater than 1. 0,and those of Xianyu 30 T60,Dika 008 and South America No. 1 were in the range of 0. 9-1. 0. The correlation between the two kinds of evaluation results was 0. 588 7 and was extremely significant. The five-level scoring method and the drought tolerance index can be used simultaneously for the evaluation of drought tolerance of maize during flowering stage. The two aspects of evaluation results showed that Guidan 0810,Zhengda 619,Xianyu 30 T60,Dika 008 and South America No. 1 were drought-tolerant varieties,among which Guidan 0810 and Zhengda 619 were extremely highly drought-tolerant varieties.

Mapping of QTLs Associated with Salt Tolerance of Maize Inbred Line at Seedling Stage

[ Objective] This study aimed to map quantitative tarit loci （QTLs） associated with salt tolerance of maize inbred line at seedling stage. [ Method! The recombinant inbred line （RIL） F7 including 171 plants were developed by single seed descent procedure from a combination, Huangzazsi × Mo17, and used to map QTLs associated with salt tolerance, based on the constructed genetic map of SSR markers. [ Resultl A linkage map consisting of 81 SSR markers loci from 10 chromosomes （ 1 428.3 cM in total length, with an average distance of 17.63 cM between two neighbouring loci） was constructed. Six QTI~ associated significantly with salt tolerance were detected at chromosomes 1,5 and 6. [ Conclusion] This study is extremely significant for better understanding sah tolerance-related genes, the genes＇ location and cloning, salt tolerance mechanism and the marker-assisted selection of salt tolerant maize.

Identifi cation of QTLs for Drought Resistance of Maize Germination and Seedling Development

In most maize-growing areas yield reductions due to drought have been observed (Frova et al., 1999; Li et al., 2003; Ribaut et al., 1997; Sari-Gorla et al., 1999). Seed germination and early seedling growth is the

Assessment of molecular markers and marker-assisted selection for drought tolerance in barley(Hordeum vulgare L.)

This review updates the present status of the field of molecular markers and marker-assisted selection(MAS),using the example of drought tolerance in barley.The accuracy of selected quantitative trait loci(QTLs),candidate genes and suggested markers was assessed in the barley genome cv.Morex.Six common strategies are described for molecular marker development,candidate gene identification and verification,and their possible applications in MAS to improve the grain yield and yield components in barley under drought stress.These strategies are based on the following five principles:(1)Molecular markers are designated as genomic‘tags’,and their‘prediction’is strongly dependent on their distance from a candidate gene on genetic or physical maps;(2)plants react differently under favourable and stressful conditions or depending on their stage of development;(3)each candidate gene must be verified by confirming its expression in the relevant conditions,e.g.,drought;(4)the molecular marker identified must be validated for MAS for tolerance to drought stress and improved grain yield;and(5)the small number of molecular markers realized for MAS in breeding,from among the many studies targeting candidate genes,can be explained by the complex nature of drought stress,and multiple stress-responsive genes in each barley genotype that are expressed differentially depending on many other factors.

Effects of Drought Stress and Re-watering on Growth and Yield of Different Maize Varieties at Tasseling Stage

[Objective] The aim was to explore the response of different maize vari- eties in Guangxi to drought stress and re-watering at tasseling stage, so as to pro- vide reference for study on drought resistance mechanism, breeding of drought-re- sistant varieties and selection of maize varieties for fall sowing. [Method] At the tasseling stage, total five degrees of drought stress （4, 8, 12, 16 and 20 d） and corresponding re-watering after drought stress were simulated by a tub planting ex- periment in greenhouse for five different maize varieties （Guidan 0810, Dika 008, Zhengda 619, Chenyu 969, Guidan 901）. Normal watering was set as the control. Sampling was carried out on Day 1 after drought stress and on Day 15 after re- watering, and the secondary root number, maximum root length, green leaf number, root dry weight and shoot dry weight were measured. At the harvest time, the ear yield per plant was measured. With yield as the basis, the drought resistance coef- ficient and drought resistance index were calculated. Cluster analysis was conducted for drought resistance coefficient. [Result] The shoot dry weight, root dry weight, secondary root number, maximum root length and green leaf number of maize in the treatment groups decreased compared with those in the control group. The ratio of each index between the treatment and control groups declined with the extension of drought stress. After re-watering, the re-growth amount of each index all de- creased as the stress time prolonged. Post-re-watering over compensation effect oc- curred in none of the indices except the maximum root length, after 4 days of drought stress. Under drought stress, the reductions of all the indices of Guidan 0810, Dika 008 and Zhengda 619 were smaller than those of Chenyu 969 and Guidan 901. After re-watering, the re-growth abilities of Guidan 0810, Dika 008 and Zhengda 619 were stronger than those of Chenyu 969 and Guidan 901. The drought resistance coefficients and drought resistance indexes of Guidan 0810, Dika 008 and Zhengda 619 were all greater than those of Chenyu 969 and Guidan 901. The results of drought resistance coefficient cluster analysis showed that the five maize varieties were classified into two groups： Guidan 0810, Dika 008 and Zheng- da 619 had strong drought resistance, while Guidan 901 and Chenyu 969 had weak drought resistance. [Conclusion] The root and shoot growth of Guidan 0810, Dika 008 and Zhengda 619 was slightly affected by drought stress during the tasseling period, and they restored the growth rapidly after re-watering, thus ensuring high biomass and yield. Therefore, Guidan 0810, Dika 008 and Zhengda 619 can be promoted as drought-resistant autumn maize varieties in Guangxi.

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.

Evaluation and Quantitative Inheritance of Several Drought-Relative Traits in Maize

The authors evaluated 57 parental inbred lines of maize hybrids disseminated in Southwest China for drought tolerance under drought-stressed and well-watered conditions. Multiple regression analyses between drought tolerant coefficients of the grain yield per plant and 15 morphological and physiological traits measured from a subset of 12 selected lines, identified traits 1 and 5, which were important for drought tolerance, at the seedling and reproductive stages respectively. Gene effects, combining abilities, and heritabilities of these traits were estimated using generation mean and diallel cross methods. Dominance effect was more important than additive effect for the plant height, anthesis-silking interval （ASI）, root weight, and the grain yield per plant, whereas, they were about equal for the leaf emergence rate. The variances of special combining ability （SCA） were about double that of the general combining ability （GCA） for plant height, ASI and grain yield per plant, although they were about equal for leaf emergence rate and root weight. Narrow sense heritabilities of the five traits for the reproductive stage were not high （12.8-29.6%）, although broad sense heritabilities for plant height, ASI, and grain yield were as high as 70-85%. A segregating population consisting of 183 F2 plants from the cross N87-1 （drought tolerant） × 9526 （susceptible）, was genotyped at 103 SSR loci and the F2：4 families were evaluated under two water regimes. Twelve quantitative trait loci （QTLs） （two for plant height, five for ASI, four for root biomass, and one for grain yield） were identified, most of which had overdominant gene action. Some chromosomal regions, such as those linked to markers umcl051 （bin 4.08）, umc2881 （bin 4.03）, and phi034 （bin 7.02）, had overlapping QTLs.

QTL Mapping for Drought Tolerance at Stages of Germination and Seedling in Wheat (Triticum aestivum L.) Using a DH Population

Drought is a major constraint in many wheat( Triticum aestivum L.) production regions. Quantitative trait loci (QTLs) conditioning drought tolerance at stages of germination and seedling in wheat were identified in a double haploid (DH) population derived from the cross, Hanxuan10×Lumai14, using amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers. Interval mapping analysis revealed that QTLs for drought tolerance at germination stage were located on chromosomes 1B, 2B, 5A, 6B, 7A and 7B, respectively, and the most effective QTL was mapped on chromosome 2B, explaining 27.2% of phenotypic variance. The QTLs for drought tolerance at seedling stage were located on 1B, 3B and 7B, respectively, and the most effective QTL was mapped on chromosome 3B, explaining 21.6% of phenotypic variance. Their positions were different from those of QTLs conferring drought tolerance at germination stage, indicating that drought tolerance at germination stage and seedling stage was controlled by different loci. Most of the identified QTLs explained 18% or more of phenotypic variance for drought tolerance at germination and seedling stage, and would be useful in future for marker assisted selection programs and cultivar improvement.

Differential Gene Expression in Response to Drought Stress in Maize Seedling

To provide the useful information for the choice of molecular marker used in marker-assisted selection of drought tolerance, it is necessary to find out more candidate genes and fulfill the information gaps in gene expression regulation under drought stress. In this study, we isolated four differentially expressed cDNA fragments from leaves of a droughttolerant inbred line by suppression subtractive hybridization and reverse Northern hybridization, and validated their differential expression patterns among six inbred lines with different drought tolerance in response to drought stress by quantitative real-time PCR. Sequence similarity analysis indicated that two of four differentially expressed cDNA showed homology to gene DegP encoding trypsin-like serine protease, and gene PGAM-i encoding cofactor-independent phosphoglyceromutase, respectively. Expressions of the genes corresponding to four cDNA fragments was decreased at 6 h after drought stress treatment in most of the six inbred lines, and then returned to the control level with further stress in three of the tolerant inbred lines. The expression of the gene PGAM-i and the genes corresponding to fragments E4 and F4 were increased to a high level in tolerant inbred line 81565. In the two drought-sensitive inbred lines （Dan340 and ES40）, the expression of these genes was still down-regulated. The probable mechanisms of these genes in response to drought stress were discussed. These results indicated that the drought-tolerant inbred lines upregulated the expression of the drought-tolerant candidate genes, in contrast, drought-sensitive inbred lines downregulated the expression of the genes.

Identification of Functional Genetic Variations Underlying Drought Tolerance in Maize Using SNP Markers

Single nucleotide polymorphism （SNP） is a common form of genetic variation and popularly exists in maize genome. An Illumina GoldenGate assay with 1 536 SNP markers was used to genotype maize inbred lines and identified the functional genetic variations underlying drought tolerance by association analysis. Across 80 lines, 1 006 polymorphic SNPs （65.5% of the total） in the assay with good call quality were used to estimate the pattern of genetic diversity, population structure, and familial relatedness. The analysis showed the best number of fixed subgroups was six, which was consistent with their original sources and results using only simple sequence repeat markers. Pairwise linkage disequilibrium （LD） and association mapping with phenotypic traits investigated under water-stressed and well-watered regimes showed rapid LD decline within 100–500 kb along the physical distance of each chromosome, and that 29 SNPs were associated with at least two phenotypic traits in one or more environments, which were related to drought-tolerant or drought-responsive genes. These drought-tolerant SNPs could be converted into functional markers and then used for maize improvement by marker-assisted selection.

Positional cloning and characterization reveal the role of a miRNA precursor gene ZmLRT in the regulation of lateral root number and drought tolerance in maize

Lateral roots play essential roles in drought tolerance in maize(Zea mays L.). However, the genetic basis for the variation in the number of lateral roots in maize remains elusive. Here, we identified a major quantitative trait locus(QTL),q LRT5-1, controlling lateral root number using a recombinant inbred population from a cross between the maize lines Zong3(with many lateral roots) and 87-1(with few lateral roots).Fine-mapping and functional analysis determined that the candidate gene for qLRT5-1,ZmLRT, expresses the primary transcript for the microRNA miR166a. ZmLRT was highly expressed in root tips and lateral root primordia,and knockout and overexpression of ZmLRT increased and decreased lateral root number,respectively. Compared with 87-1, the ZmLRT gene model of Zong3 lacked the second and third exons and contained a 14 bp deletion at the junction between the first exon and intron,which altered the splicing site. In addition,ZmLRT expression was significantly lower in Zong3 than in 87-1, which might be attributed to the insertions of a transposon and over large DNA fragments in the Zong3 ZmLRT promoter region. These mutations decreased the abundance of mature miR166a in Zong3, resulting in increased lateral roots at the seedling stage.Furthermore, miR166a post-transcriptionally repressed five development-related class-Ⅲ homeodomain-leucine zipper genes. Moreover,knockout of ZmLRT enhanced drought tolerance of maize seedlings. Our study furthers our understanding of the genetic basis of lateral root number variation in maize and highlights ZmLRT as a target for improving drought tolerance in maize.

Piriformospora indica confers drought tolerance on Zea mays L.through enhancement of antioxidant activity and expression of drought-related genes

Drought stress is one of the most severe environmental constraints to plant growth and crop productivity. Plant growth is greatly affected by drought stress, and plants, to survive,adapt to this stress by invoking different pathways. Piriformospora indica, a root-colonizing endophytic fungus of Sebacinales, promotes plant growth and confers resistance to biotic and abiotic stresses, including drought stress, by affecting the physiological properties of the host plant. The fungus strongly colonizes the roots of maize(Zea mays L.) and promotes shoot and root growth under both normal growth conditions and drought stress. We used polyethylene glycol(PEG-6000) to mimic drought stress and found that root fresh and dry weight, leaf area, SPAD value, and leaf number were increased in P. indica-colonized plants.The antioxidative activities of catalases and superoxide dismutases were upregulated within 24h in the leaves of P. indica-colonized plants. Drought-related genes DREB2A, CBL1,ANAC072, and RD29A were upregulated in drought-stressed leaves of P. indica-colonized plants. Furthermore, after drought treatment, proline content increased, whereas accumulation of malondialdehyde(MDA), an indicator of membrane damage, decreased in P. indica-colonized maize. We conclude that P. indica-mediated plant protection against the detrimental effects of drought may result from enhanced antioxidant enzyme activity,proline accumulation, and expression of drought-related genes and lower membrane damage in maize plants.

Advances in deciphering salt tolerance mechanism in maize

Maize(Zea mays L.)is a global cereal crop whose demand is projected to double by 2050.Along with worsening of farmland salinization,salt stress has become a major environmental threat to the sustainability of maize production worldwide.Accordingly,there is an urgent need to decipher salt-tolerant mechanisms and facilitate the breeding of salt-tolerant maize.As salt tolerance is a complex trait regulated by multiple genes,and maize germplasm varies widely in salt tolerance,efforts have been devoted to the identification and application of quantitative-trait loci(QTL)for salt tolerance.QTL associated with ion regulation,osmotic tolerance,and other aspects of salt tolerance have been discovered using genomewide association studies(GWAS),linkage mapping,and omics-based approaches.This review highlights recent advances in the molecular-level understanding of salt stress response in maize,in particular in(a)the discovery of salt-tolerance QTL,(b)the mechanisms of salt tolerance,(c)the development of salttolerant maize cultivars,and(d)current challenges and future prospects.

Phenotyping of Hybrid Maize (<i>Zea mays</i>L.) at Seedling Stage under Drought Condition

Drought is envisaged as the greatest demolishing natural impacts throughout the world since it has observed extensive place of agronomical land sterile almost the world. It’s the significant crop output-limiting producer, and elaborated learning of its result on plant enhancement dictation is diametrical. At present, drought tolerant hybrid maize has been trying to induce Bangladesh especially drought affected zone to identify the drought endurance maize genotypes. Consequently, a feasible pot study of 49 hybrid maize genotypes were directed to determine an adequate drought level to promote aliment and promotion of maize plant below the water stress conditions with treatment (control and drought) and three replications. The data were received after 35 days of sowing using appropriate procedures. Specially, the stomata were collected by the white transparent nail polish from the lower part of leaves. Descriptive statistic of the all traits like percentage of SPAD, leaf rolling (LR), maximum root length (MRL), maximum shoot length (MSL), root dry matter (RDM), shoot dry matter (SDM), length of stomata (LS), width of stomata (WS), thickness of stomata (TS), total dry matter (TDM) and ANOVA for control and drought condition individually showed significant (P < 0.05) variations among the germplasm for their genotypes, treatment and interaction. The first fourth principal components (PCs) narrated about 82.0% of the total variation. Cluster analysis placed the 49 hybrid into 6 main groups among those cluster;groups five showed the maximum number mean value of traits. The highest positive relationship was obtained from TS, WS, RDM, SDM and TDM traits by forming genotype-traits bi-plot of 11traits of 49 genotypes. After analyzing, it is explicit that G18 (CML-80 × IPB911-16) and G22 (CZI-04 × IPB911-16) were the most tolerant hybrids maize genotypes and very susceptible hybrids maize genotypes were G16 (P-12 × CML487), G34 (CML-32 × PB911-16) and G37 (P-33 × CML487). It is expected that the higher expression of considered traits might be obligate for better yield under drought stress.

Potentials of Arbuscular Mycorrhiza Fungus in Tolerating Drought in Maize (<i>Zea mays</i>L.)

Maize is one of the most important cereal crops widely grown for food, feed, and fodder/forage throughout the world in a range of agroecological environments. Drought stress continues to haunt the maize farmers across south western part of Nigeria, thereby leading to low quantity of this essential staple food in the market. Efforts have been made to enhance the growths and yields in maize by investigating the influence of Arbuscular mycorrhizal fungus (Gigaspora gigantea) on the tolerance of maize to drought stress. The experiment was conducted in the teaching and research farm of Babcock University, Ilishan-Remo, Nigeria. The experiment was laid out in a complete randomized design with four replicates. Data were collected on eight morphological drought related characters. The objective of this research work was to evaluate the morpho-agronomic responses and potential of Gigaspora gigantea colonization in maize drought tolerance, and also to select parents in maize breeding for improved yield related components. The combined analysis of variance showed significant (P 0.05) treatment effect on majority of the traits evaluated. The treatments of Arbuscular Mycorrhiza Fungus (AMF) produced significant higher growth related traits suggesting that AMF treated plants had higher potential in influencing the tolerance to drought. Accession 3 was considered best for most of the traits studied and can be selected as parents in maize breeding for yield related components.

Integrated linkage mapping and genome-wide association study to dissect the genetic basis of zinc deficiency tolerance in maize at seedling stage

Zinc(Zn)deficiency is the most widespread micronutrient deficiency,affecting yield and quality of crops worldwide.Identifying genes associated with Zn-deficiency tolerance in maize is a basis for elucidating its genetic mechanism.A K22×CI7 recombinant inbred population consisting of 210 lines and an association panel of 508 lines were used to identify genetic loci influencing Zn-deficiency tolerance.Under-Zn and-Zn/CK conditions,15 quantitative trait loci(QTL)were detected,each explaining 5.7%-12.6%of phenotypic variation.Sixty-one significant single-nucleotide polymorphisms(SNPs)were identified at P<10^(-5)by genome-wide association study(GWAS),accounting for 5%-14%of phenotypic variation.Among respectively 198 and 183 candidate genes identified within the QTL regions and the 100-kb regions flanking these significant SNPs,12 were associated with Zn-deficiency tolerance.Among these candidate genes,four genes associated with hormone signaling in response to Zn-deficiency stress were co-localized with QTL or SNPs,including the genes involved in the auxin(ZmARF7),and ethylene(ZmETR5,ZmESR14,and ZmEIN2)signaling pathways.Three candidate genes were identified as being responsible for Zn transport,including ZmNAS3 detected by GWAS,ZmVIT and ZmYSL11 detected by QTL mapping.Expression of ZmYSL11 was up-regulated in Zn-deficient shoots.Four candidate genes that displayed different expression patterns in response to Zn deficiency were detected in the regions overlapping peak GWAS signals,and the haplotypes for each candidate gene were further analyzed.

Improving Drought Tolerance of Rice by Designed QTL Pyramiding

Drought is the most important factor limiting rice yields in the rainfed areas of Asia. To overcome the problem, we developed a new strategy 'designed QTL pyramiding' to more effi ciently develop drought tolerant (DT)