Population genomic analysis reveals key genetic variations and the driving force for embryonic callus induction capability in maize

Genetic transformation has been an effective technology for improving the agronomic traits of maize.However,it is highly reliant on the use of embryonic callus(EC)and shows a serious genotype dependence.In this study,we performed genomic sequencing for 80 core maize germplasms and constructed a high-density genomic variation map using our newly developed pipeline(MQ2Gpipe).Based on the induction rate of EC(REC),these inbred lines were categorized into three subpopulations.The low-REC germplasms displayed more abundant genetic diversity than the high-REC germplasms.By integrating a genome-wide selective signature screen and region-based association analysis,we revealed 95.23 Mb of selective regions and 43 REC-associated variants.These variants had phenotypic variance explained values ranging between 21.46 and 49.46%.In total,103 candidate genes were identified within the linkage disequilibrium regions of these REC-associated loci.These genes mainly participate in regulation of the cell cycle,regulation of cytokinesis,and other functions,among which MYB15 and EMB2745 were located within the previously reported QTL for EC induction.Numerous leaf area-associated variants with large effects were closely linked to several REC-related loci,implying a potential synergistic selection of REC and leaf size during modern maize breeding.

A novel semi-dominant allele of the transmembrane NAC transcription factor ZmNTL2 reduces the size of multiple maize organs

Maize plant architecture influences planting density and,in turn,grain yield.Most of the plant architecture-related traits can be described as organ size.We describe a miniature maize mutant,Tiny plant 4(Tip4),which exhibits reduced size of multiple organs and exhibits a semi-dominant monofactorial inheritance characteristic.Positional cloning confirmed that a 4-bp deletion in the NAC TF with transmembrane motif 1-Like(NTL)gene ZmNTL2,denoted as ZmNTL2^(Δ),confers the Tip4 mutation.qRT-PCR showed that ZmNTL2 was expressed in all tested tissues.ZmNTL2 functions as a transcriptional activator and is located in both the nucleus and biomembranes.The mutation does not affect the mRNA abundance of ZmNTL2 locus,but it does result in the loss of transmembrane domain and confines the ZmNTL2^(Δ)protein to the nucleus.Knocking out ZmNTL2 has no effect on maize organ size development,indicating that the 4-bp deletion might be a gain-of-function mutation in organ size regulation.Combining transcriptome sequencing with cytokinin and auxin content determination suggests that the decreased organ size may be possibly mediated by changes in hormone homeostasis.

Genetic analysis and candidate gene identification of salt tolerancerelated traits in maize

Soil salinization poses a threat to maize production worldwide,but the genetic mechanism of salt tolerance in maize is not well understood.Therefore,identifying the genetic components underlying salt tolerance in maize is of great importance.In the current study,a teosinte-maize BC2F7 population was used to investigate the genetic basis of 21 salt tolerance-related traits.In total,125 QTLs were detected using a high-density genetic bin map,with one to five QTLs explaining 6.05–32.02%of the phenotypic variation for each trait.The total phenotypic variation explained(PVE)by all detected QTLs ranged from 6.84 to 63.88%for each trait.Of all 125 QTLs,only three were major QTLs distributed in two genomic regions on chromosome 6,which were involved in three salt tolerance-related traits.In addition,10 pairs of epistatic QTLs with additive effects were detected for eight traits,explaining 0.9 to 4.44%of the phenotypic variation.Furthermore,18 QTL hotspots affecting 3–7 traits were identified.In one hotspot(L5),a gene cluster consisting of four genes(ZmNSA1,SAG6,ZmCLCg,and ZmHKT1;2)was found,suggesting the involvement of multiple pleiotropic genes.Finally,two important candidate genes,Zm00001d002090 and Zm00001d002391,were found to be associated with salt tolerance-related traits by a combination of linkage and marker-trait association analyses.Zm00001d002090 encodes a calcium-dependent lipid-binding(CaLB domain)family protein,which may function as a Ca^(2+)sensor for transmitting the salt stress signal downstream,while Zm00001d002391 encodes a ubiquitin-specific protease belonging to the C19-related subfamily.Our findings provide valuable insights into the genetic basis of salt tolerance-related traits in maize and a theoretical foundation for breeders to develop enhanced salt-tolerant maize varieties.

Multi-omics analysis reveals the pivotal role of phytohormone homeostasis in regulating maize grain water content

Grain water content(GWC)is a key determinant for mechanical harvesting of maize(Zea mays).In our previous research,we identified a quantitative trait locus,qGWC1,associated with GWC in maize.Here,we examined near-isogenic lines(NILs)NILL and NILH that differed at the qGWC1 locus.Lower GWC in NILL was primarily attributed to reduced grain water weight(GWW)and smaller fresh grain size,rather than the accumulation of dry matter.The difference in GWC between the NILs became more pronounced approximately 35 d after pollination(DAP),arising from a faster dehydration rate in NILL.Through an integrated analysis of the transcriptome,proteome,and metabolome,coupled with an examination of hormones and their derivatives,we detected a marked decrease in JA,along with an increase in cytokinin,storage forms of IAA(IAA-Glu,IAA-ASP),and IAA precursor IPA in immature NILL kernels.During kernel development,genes associated with sucrose synthases,starch biosynthesis,and zein production in NILL,exhibited an initial up-regulation followed by a gradual down-regulation,compared to those in NILH.This discovery highlights the crucial role of phytohormone homeostasis and genes related to kernel development in balancing GWC and dry matter accumulation in maize kernels.

Phenotypic Characterization and QTL/Gene Identification for Internode Number and Length Related Traits in Maize

Internode number and length are the foundation to constitute plant height, ear height and the above-ground spatial structure of maize plant. In this study, segregating populations were constructed between EHel with extremely low ear height and B73. Through the SNP-based genotyping and phenotypic characterization, 13 QTL distributed on the chromosomes (Chrs) of Chr1, Chr2, Chr5-Chr8 were detected for four traits of internode no. above ear (INa), average internode length above ear (ILaa), internode no. below ear (INb), and average internode length below ear (ILab). Phenotypic variation explained (PVE) by a single QTL ranged from 6.82% (qILab2-2) to 12.99% (qILaa5). Zm00001d016823 within the physical region of qILaa5, the major QTL for ILaa with the largest PVE was determined as the candidate through the genomic annotation and sequence alignment between EHel and B73. Product of Zm00001d016823 was annotated as a WEB family protein homogenous to At1g75720. qRT-PCR assay showed that Zm00001d016823 highly expressed within the tissue of internode, exhibiting statistically higher expression levels among internodes of IN4 to IN7 in EHel than those in B73 (P Zm00001d016823 might provide novel insight into molecular mechanism beyond phytohormones controlling internode development in maize.

Stability of QTL Across Environments and QTL-by-Environment Interactions for Plant and Ear Height in Maize

Better understanding of genotype-by-environment interaction （GEI） is expected to provide a solid foundation for genetic improvement of crop productivity especially under drought-prone environments. To elucidate the genetic basis of the plant and ear height, 2 F2：3 populations were derived from the crosses of Qi 319 × Huangzaosi （Q/H） and Ye 478 × Huangzaosi （Y/H） with 230 and 235 families, respectively, and their parents were evaluated under 3 diverse environments in Henan, Beijing, and Xinjiang, China during the year of 2007 and 2008, and all the lines were also evaluated under water stress environment. The mapping results showed that a total of 21 and 12 QTLs were identified for plant height in the Q/H and Y/H population, respectively, and 24 and 13 QTLs for ear height, respectively. About 56 and 73% of the QTLs for 2 traits did not present significant QTL-by-environment interaction （QE1） in the normal joint analyses for Q/H and Y/H population, respectively, and about 73% of the QTLs detected did not show significant QEI according to joint analyses for stress condition in Q/H. Most of the detected major QTLs exhibited high stability across different environments. Besides, several major QTLs were detected with large and consistent effect under normal condition （Chr. 6 and 7 in Q/H; Chr. 1, 3 and 9 in Y/H）, or across 2 water regimes （Chr. 1, 8 and 10 for in Q/H）. There were several constitutive QTLs （3 for Q/H and 1 for Y/H） with no or minor QTL-by-environment for the 2 populations. Finally, we found several genomic regions （Chr. 1, 10, etc.） to be co-located across the populations, which could provide useful reference for genetic improvement of these traits in maize breeding programs. Comparative genomic analysis revealed that 3 genes/genetic segments associated with plant height in rice were orthologous to these 3 identified genomic regions carrying the major QTLs for plant and ear height on Chr. 1, 6, and 8, respectively.

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.

Genetic Diversity of Maize Populations Developed by Two Kinds of Recurrent Selection Methods Investigated with SSR Markers

Two cycles of biparental mass selection （MS） and one cycle of half-sib-S3 family combining selection （HS-S3） for yield were carried out in 2 synthetic maize populations P4C0 and P5C0 synchronously. The genetic diversity of 8 maize populations, including both the basic populations and their developed populations, were evaluated by 30 SSR primers. On the 30 SSR loci, a total of 184 alleles had been detected in these populations. At each locus, the number of alleles varied from 2 to 14, with an average of 6.13. The number and ratio of polymorphic loci in both the basic populations were higher than those of their developed populations, respectively. There was nearly no difference after MS but decreased after HS-S3 in both the basic populations in the mean gene heterozygosity. The mean genetic distance changed slightly after MS but decreased in a bigger degree after HS-S3 in both the basic populations. Analyses on the distribution of genetic distances showed that the ranges of the genetic distance were wider after MS and most of the genetic distances in populations developed by HS-S3 were smaller than those in both the basic populations. The number of genotypes increased after MS but decreased after HS-S3 in both the basic populations. The genetic diversity of intra-population was much more than genetic diversity of inter-population in both the basic populations. All these indexes demonstrated that the genetic diversity of populations after MS was similar to their basic populations, and the genetic diversity was maintained during MS, whereas the genetic diversity of populations decreased after HS-S3. This result indicated that heterogeneity between some of the individuals in the developed populations increased after MS, whereas the populations become more homozygotic after HS-S3.

Dissection of the genetic architecture for tassel branch number by QTL analysis in two related populations in maize

Tassel branch number （TBN） is the principal component of maize tassel inflorescence architecture and is a typical quan- titative trait controlled by multiple genes. The main objective of this research was to detect quantitative trait loci （QTLs） for TBN. The maize inbred line SICAU1212 was used as the common parent to develop BC1S1 and recombinant inbred line （RIL） populations with inbred lines 3237 and B73, respectively. The two related populations consisted of 123 and 238 lines, respectively. Each population was grown and phenotyped for TBN in two environments. Eleven QTLs were detected in the BC1S1 population, located on chromosomes 2, 3, 5, and 7, accounted for 4.45-26.58% of the phenotypic variation. Two QTLs （qB11Jtbn2-1, qB12Ctbn2-1, qBJtbn2-1; q11JBtbn5-1, qB12Ctbn5-1, qBJtbn5-1） that accounted for more than 10% of the phenotypic variation were identified. Three QTLs located on chromosomes 2, 3 and 5, exhibited stable expres- sion in the two environments. Ten QTLs were detected in the RIL population, located on chromosomes 2, 3, 5, 8, and 10, accounted for 2.69-13.58% of the TBN variation. One QTL （qR14Dtbn2-2） explained 〉10% of the phenotypic variation. One common QTL （qB12Ctbn2-2, qR14Dtbn2-2, qRJtbn2-2） was detected between the two related populations. Three pairs of epistatic effects were identified between two loci with or without additive effects and accounted for 1.19-4.26% of the phenotypic variance. These results demonstrated that TBN variation was mainly caused by major effects, minor effects and slightly modified by epistatic effects. Thus, identification of QTL for TBN may help elucidate the genetic basis of TBN and also facilitate map-based cloning and marker-assisted selection （MAS） in maize breeding programs.

The Effects of Low Phosphorus Stress on Morphological and Physiological Characteristics of Maize (Zea mays L.) Landraces

A field trial was conducted to investigate main morphological and physiological changes of different maize landraces to low-P stress at the stage of seedling. P-deficiency significantly decreased root volume, total leaf area, and plant dry weight, but greatly increased density of root hairs and root top ratio. In addition, P-deficiency induced the significant enhancement of phosphorus utilization efficiency and the amount of proline, malondialdehye （MDA）, acid phosphatase （APase）, peroxidase （POD） and superoxide dismutase （SOD）, but the significant reduction of P uptake and soluable protein content. Since P-deficiency had smaller effects on the P-tolerant maize landraces DP-44, DP-32 and DP-33 as compared with P-sensitive landraces DP-29 and DP-24, it was demonstrated that differences of tolerance to P-deficiency existed among different maize landraces. The results based on the correlation analysis showed that the economic yield of maize landraces had relationships with their morphological and physiological characteristics under P-deficiency.

Comparative Analysis of Genetic Diversity in Landraces of Waxy Maize from Yunnan and Guizhou Using SSR Markers

Waxy maize landraces are abundant inYunnan and Guizhou of China. Genetic diversity of waxy maize landraces from Yunnan and Guizhou were analyzed using SSR markers. We screened 38 landraces with 50 primers that generated 3 to 6 polymorphic bands, with an average of 4.13 bands. Shannon＇s information indices for genetic diversity of the 14 waxy maize landraces from Yunnan varied from 4.9571 to 42.1138 and averaged 26.5252; Shannon＇s information indices for genetic diversity of the 24 waxy maize landraces from Guizhou varied from 22.0066 to 40.6320 and averaged 32.3156. For the 14 waxy maize landraces from Yunnan, the within-landrace genetic diversity accounted for 45.40% and the among-landrace genetic diversity accounted for 54.60% of the total genetic diversity observed. For the 24 waxy maize landraces from Guizhou, the within-landrace genetic diversity accounted for 50.76% and the among-landrace genetic diversity accounted for 49.24% of the total observed. Some individual landraces possessed as much as 96.86% of the total genetic diversity occurring among landraces within origins. Differentiation between geographic origins accounted for only 3.14% of the total genetic diversity. Both Yunnan and Guizhou would be the diversity centers and the original centers of waxy maize.

Fosmid library construction and screening for the maize mutant gene Vestigial glume 1

The maize mutant gene Vestigial glume 1(Vg1) has been fine-mapped to a narrow region by map-based cloning and the candidate gene for Vg1 spanned 19.5 kb. Here we report Vg1 genomic fosmid library construction and screening. The fosmid library of Vg1 consisted of574,000 clones with an average insert size of 36.4 kb, representing 7.9-fold coverage of the maize genome. Fosmid stability assays indicated that clones were stable during propagation in the fosmid system. Using Vg1 candidate gene-specific primers, a positive clone was successfully identified. This discovery will pave the way for identifying the function of Vg1 in maize development.

Mapping Quantitative Trait Loci Associated with Photoperiod Sensitivity in Maize(Zea mays L.)

Photoperiod sensitivity in maize plays an essential role in utilizing tropic and sub-tropic germplasm to temperate areas. This study aims to identify and map the QTLs responsible for the characteristics measuring photoperiod sensitivity, days from planting to silking （SD）, photoperiod response coefficient of silking （PRC）, and anthesis-silking interval （ASI）. Using the population derived from Zheng 58, photoperiod-insensitive parent, and Ya 8701, photoperiod-sensitive parent, a linkage map was constructed with 93 single sequence repeat （SSR） markers. Phenotyping of 296 F2-3 families of the population in replicated-field test was conducted in both long-day （Beijing, China） and short-day （Sichuan, China） conditions. Ten QTLs were identified to be associated with the SD and ASI on chromosomes 3, 4, 6, 8, and 10 in the longday conditions, and 11 QTLs were detected to be related to the SD and ASI on chromosomes 2, 3, 4, 5, 6, 8, and 10 in the short-day conditions, respectively. A QTL associated with the PRC as a major effect in the long-day conditions located in the same position as the QTL related to the SD and ASI in the map, and was on chromosome 10 linked with marker bnlg1655. Using these QTLs in the marker-assisted selection, the photoperiod sensibility could be reduced by selection of the alleles responsible for the SD, PRC, and ASI in breeding programs.

Zea mays(L.) P1 locus for cob glume color identified as a post-domestication selection target with an effect on temperate maize genomes

Artificial selection during domestication and post-domestication improvement results in loss of genetic diversity near target loci. However, the genetic locus associated with cob glume color and the nature of the genomic pattern surrounding it was elusive and the selection effect in that region was not clear. An association mapping panel consisting of 283 diverse modern temperate maize elite lines was genotyped by a chip containing over 55,000 evenly distributed SNPs. Ten-fold resequencing at the target region on 40 of the panel lines and 47 tropical lines was also undertaken. A genome-wide association study(GWAS) for cob glume color confirmed the P1 locus, which is located on the short arm of chromosome 1, with a-log10 P value for surrounding SNPs higher than the Bonferroni threshold(α/n, α < 0.001) when a mixed linear model(MLM) was implemented. A total of 26 markers were identified in a 0.78 Mb region surrounding the P1 locus, including 0.73 Mb and 0.05 Mb upstream and downstream of the P1 gene, respectively. A clear linkage disequilibrium(LD) block was found and LD decayed very rapidly with increasing physical distance surrounding the P1 locus. The estimates of π and Tajima's D were significantly(P < 0.001) lower at both ends compared to the locus. Upon comparison of temperate and tropical lines at much finer resolution by resequencing(180-fold finer than chip SNPs), a more structured LD block pattern was found among the 40 resequenced temperate lines. All evidence indicates that the P1 locus in temperate maize has not undergone neutral evolution but has been subjected to artificial selection during post-domestication selection or improvement. The information and analytical results generated in this study provide insights as to how breeding efforts have affected genome evolution in crop plants.

Characterization of the imprinting and expression patterns of ZAG2 in maize endosperm and embryo

ZAG2 has been identified as a maternally expressed imprinted gene in maize endosperm.Our study revealed that paternally inherited ZAG2 alleles were imprinted in maize endosperm and embryo at 14 days after pollination(DAP), and consistently imprinted in endosperm at 10, 12, 16, 18, 20, 22, 24, 26, and 28 DAP in reciprocal crosses between B73 and Mo17. ZAG2 alleles were also imprinted in reciprocal crosses between Zheng 58 and Chang7-2 and between Huang C and 178. ZAG2 alleles exhibited differential imprinting in hybrids of 178 × Huang C and B73 × Mo17, while in other hybrids ZAG2 alleles exhibited binary imprinting. The tissue-specific expression pattern of ZAG2 showed that ZAG2 was expressed at a high level in immature ears, suggesting that ZAG2 plays important roles in not only kernel but ear development.

Differential Expression of MicroRNAs in Response to Drought Stress in Maize

Drought is one of the major abiotic stresses that limit maize productivity. Apart from the principal transcriptional regulation, post-transcriptional regulation mediated by microRNAs appears to be the prevalent response of plants to abiotic stress. In this study, the differential expression of microRNAs in the previously evaluated drought-tolerant inbred lines R09 under drought stress was detected by microarray hybridization. The target genes of the differentially-expressed microRNAs were predicted by bioinformatics software WMD3 for plant target gene prediction. The possible regulation of the differentially-expressed microRNAs as well as their target genes in maize response to drought stress was analysed according to Gene Ontology. Sixty-eight microRNAs in 29 microRNA families were detected to be differentially expressed in the seedling of the drought-tolerant inbred line R09, accounting for 5.97% of the total number of the probes. The expression profiles were different between the two time points of the drought stress. The functions of the genes targeted by the differentially-expressed microRNAs involve multiple physiological and biochemical pathways of response to abiotic stress, such as transcription regulation, metabolism, signal transduction, hormone stimulation, and transmembrane transport. Under drought stress, the differential expression of microRNAs regulates the expression of their target genes, resulting in multiple responses of physiological and biochemical pathways relative to drought tolerance of maize, miR156, miR159 and miR319 families may play more important roles. The different members of the same family may play similar regulation effects in most cases.

Genome-wide association study and genomic prediction of Fusarium ear rot resistance in tropical maize germplasm

Fusarium ear rot(FER)is a destructive maize fungal disease worldwide.In this study,three tropical maize populations consisting of 874 inbred lines were used to perform genomewide association study(GWAS)and genomic prediction(GP)analyses of FER resistance.Broad phenotypic variation and high heritability for FER were observed,although it was highly influenced by large genotype-by-environment interactions.In the 874 inbred lines,GWAS with general linear model(GLM)identified 3034 single-nucleotide polymorphisms(SNPs)significantly associated with FER resistance at the P-value threshold of 1×10^(-5),the average phenotypic variation explained(PVE)by these associations was 3%with a range from 2.33%to 6.92%,and 49 of these associations had PVE values greater than 5%.The GWAS analysis with mixed linear model(MLM)identified 19 significantly associated SNPs at the P-value threshold of 1×10^(-4),the average PVE of these associations was 1.60%with a range from 1.39%to 2.04%.Within each of the three populations,the number of significantly associated SNPs identified by GLM and MLM ranged from 25 to 41,and from 5 to 22,respectively.Overlapping SNP associations across populations were rare.A few stable genomic regions conferring FER resistance were identified,which located in bins 3.04/05,7.02/04,9.00/01,9.04,9.06/07,and 10.03/04.The genomic regions in bins 9.00/01 and 9.04 are new.GP produced moderate accuracies with genome-wide markers,and relatively high accuracies with SNP associations detected from GWAS.Moderate prediction accuracies were observed when the training and validation sets were closely related.These results implied that FER resistance in maize is controlled by minor QTL with small effects,and highly influenced by the genetic background of the populations studied.Genomic selection(GS)by incorporating SNP associations detected from GWAS is a promising tool for improving FER resistance in maize.

Genetic Diversity of Two Important Groups of Maize Landraces with Same Name in China Revealed by M13 Tailed-Primer SSRs

Maize landraces White Dent and Golden Queen played a very important role in the pre-hybrid era of maize production in China. However, dozens of accessions with the same names of White Dent and Golden Queen are preserved in China National Genebank （CNG）. The present study investigated the genetic diversity of these two important groups of maize landraces, as well as the relationships within and among them. Thirty-four landrace accessions with the name of White Dent and 10 with Golden Queen preserved in CNG were fingerprinted with 52 simple sequence repeats with tailed primer M13. Summary statistics including average number of alleles per locus, gene diversity/expected heterozygosity, and observed heterozygosity were carried out using PowerMarker ver. 3.25 software. The test of Hardy-Weinberg equilibrium （HWE） and linkage disequilibrium （LD） of all the 44 maize landrace accessions were also performed by PowerMarker. We observed a significant differentiation in terms of the average number of alleles between White Dent and Golden Queen （6.44 alleles per locus in White Dent, 4.48 in Golden Queen）, while both groups of maize landraces had a relatively high but similar gene diversity （0.61 of White Dent, 0.63 of Golden Queen）. The fixation index （FST） was only 0.0044, while the percentage of loci deviated from Hardy-Weinberg equilibrium within these two groups of White Dent and Golden Queen was 32.69 and 3.92%, respectively. The rather high genetic diversity and average number of alleles per locus confirmed that both groups of landraces had a rather broad germplasm base. The extremely low fixation index showed that there was little genetic variation between White Dent and Golden Queen and the molecular variation within these two groups was remarkably high, indicating no genetic drift between White Dent and Golden Queen and suggesting different improvement approaches to these two important groups of landraces. Hardy-Weinberg equilibrium test revealed that the group of White Dent was deviated from HWE, whereas Golden Queen was under HWE.

Isolating Soil Drought-Induced Genes from Maize Seedling Leaves Through Suppression Subtractive Hybridization

In this study, a forward cDNA library was constructed by suppression subtractive hybridization using seedling leaves of CN165, a drought-tolerant maize inbred line. In the suppression subtractive hybridization （SSH） library, 672 positive clones were picked up randomly. After polymerase chain reaction （PCR） of each clone, all the single clones were sequenced. Totally 598 available sequences were obtained. After cluster analysis of the EST sequences, 80 uniESTs were obtained, among which 57 uniESTs were contigs and 23 uniESTs were singlets. The results of BLASTN showed that all the uniESTs had homologous sequences in the nr database. The BLASTX results indicated that 68 uniESTs had significant protein homology, 8 uniESTs with homology of unknown proteins and putative proteins, and 4 uniESTs without protein homology. Those drought stress-induced genes were involved in many metabolism pathways to regulate plant growth and development under drought stress.

Development of Sequence Characterized Amplified Region (SCAR) Primers for the Detection of Resistance to Sporisorium reiliana in Maize

Head smut of maize （Zea mays L.）, which was caused by Sporisorium reiliana, occurred in most of the maize growing areas of the world. The purpose of this study was to develop SCAR markers for map-based cloning of resistance genes and MAS. Two sets of BC3 progenies, one （BC3Q） derived from the cross Qi319 （resistance）×Huangzao 4 （susceptible）, the other （BC3M） from Mol7 （resistance）× Huangzao 4 （susceptible）, were generated. Huangzao 4 was the recurrent parent in both progenies. A combination of BSA （bulked segregant analysis） with AFLP （amplified fragment length polymorphism） method was applied to map the genes involving the resistance to S. reiliana, and corresponding resistant and susceptible bulks and their parental lines were used for screening polymorphic AFLP primer pairs. One fragment of PI3M61-152 was converted into SCAR （sequence charactered amplified fragment） marker S130. The marker was mapped at chromosome bin 2.09, the interval of a major QTL region previously reported to contribute to S. reiliana resistance. Furthermore, S130 was highly and facilitate map-based cloni associated with resistance to S. reiliana, and could be useful for marker-assisted selection ng of resistance genes.