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Wolfiporia cocos is an important medicinal and edible fungus that grows in association with pine trees, and its dried sclerotium has been used as a traditional medicine in China for centuries. However, the commercial production of W. cocos sclerotia is currently limited by shortages in pine wood resources. Since protein phosphatases (PPs) play significant roles in growth, signal transduction, development, metabolism, sexual reproduction, cell cycle, and environmental stress responses in fungi, the phosphatome of W. cocos was analyzed in this study by identifying PP genes, studying transcript profiles and assigning PPs to orthologous groups. Fifty-four putative PP genes were putatively identified in W. cocos genome based on homologous sequences searching using BLASTx program against the Saccharomyces cerevisiae, Fusarium graminearum, and Sclerotinia sclerotiorum databases. Based on known and presumed functions of orthologues of these PP genes found in other fungi, the putative roles of these W. cocos PPs in colonization, hyphal growth, sclerotial formation, secondary metabolism, and stress tolerance to environment were discussed in this study. And the level of transcripts from PP genes in the mycelium and sclerotium stages was also analyzed by qRT-PCR. Our study firstly identified and functional discussed the phosphatome in the medicinal and edible fungus W. cocos. The data from our study contribute to a better understanding of PPs potential roles in various cellar processes of W. cocos, and systematically provide comprehensive and novel insights into W. cocos economically important traits that could be extended to other fungi. [ABSTRACT FROM AUTHOR]

Key message: Maize kernel row number might be dominated by a set of large additive or partially dominant loci and several small dominant loci and can be accurately predicted by fewer than 300 top KRN-associated SNPs. Abstract: Kernel row number (KRN) is an important yield component in maize and directly affects grain yield. In this study, we combined linkage and association mapping to uncover the genetic architecture of maize KRN and to evaluate the phenotypic predictability using these detected loci. A genome-wide association study revealed 31 associated single nucleotide polymorphisms (SNPs) representing 17 genomic loci with an effect in at least one of five individual environments and the best linear unbiased prediction (BLUP) over all environments. Linkage mapping in three F populations identified 33 KRN quantitative trait loci (QTLs) representing 21 QTLs common to several population/environments. The majority of these common QTLs that displayed a large effect were additive or partially dominant. We found 70 % KRN-associated genomic loci were mapped in KRN QTLs identified in this study, KRN-associated SNP hotspots detected in NAM population and/or previous identified KRN QTL hotspots. Furthermore, the KRN of inbred lines and hybrids could be predicted by the additive effect of the SNPs, which was estimated using inbred lines as a training set. The prediction accuracy using the top KRN-associated tag SNPs was obviously higher than that of the randomly selected SNPs, and approximately 300 top KRN-associated tag SNPs were sufficient for predicting the KRN of the inbred lines and hybrids. The results suggest that the KRN-associated loci and QTLs that were detected in this study show great potential for improving the KRN with genomic selection in maize breeding. [ABSTRACT FROM AUTHOR]

Key message: Thirty loci for fertility restoration of pollen fertility, anther exsertion and pollen shedding to maize CMS-S were identified by GWAS. Abstract: S type cytoplasmic male sterile (CMS-S) is the main type of CMS in maize; poor understanding of the genetic architecture of fertility restoration to CMS-S is one of the reasons to impede its utility in hybrid breeding. In this study, genome-wide identification of genetic loci for fertility restoration ability to CMS-S was firstly conducted with a set of testcrossing association mapping panel in three environments. A total of 19, 3 and 8 significant loci ( P < 1.8 × 10, α = 1) for pollen fertility, anther exsertion and pollen shedding were identified, respectively, and individual locus explained up to 28.26 % of phenotypic variation. Of them, only Rf3, the main restorer-fertility gene of CMS-S, was identified for the three traits simultaneously. In addition, 83 candidate genes within the 100 kb extension regions of these loci were predicted. These results revealed that besides Rf3 multiple genetic loci and mechanisms are involved in the fertility restoration ability to CMS-S. Results in this study would provide important information for understanding the genetic architecture of fertility restoration to CMS-S in maize. [ABSTRACT FROM AUTHOR]

Key Message: Twelve major QTL in five optimal clusters and several epistatic QTL are identified for maize kernel size and weight, some with pleiotropic will be promising for fine-mapping and yield improvement. Abstract: Kernel size and weight are important target traits in maize ( Zea mays L.) breeding programs. Here, we report a set of quantitative trait loci (QTL) scattered through the genome and significantly controlled the performance of four kernel traits including length, width, thickness and weight. From the cross V671 (large kernel) × Mc (small kernel), 270 derived F families were used to identify QTL of maize kernel-size traits and kernel weight in five environments, using composite interval mapping (CIM) for single-environment analysis along with mixed linear model-based CIM for joint analysis. These two mapping strategies identified 55 and 28 QTL, respectively. Among them, 6 of 23 coincident were detected as interacting with environment. Single-environment analysis showed that 8 genetic regions on chromosomes 1, 2, 4, 5 and 9 clustered more than 60 % of the identified QTL. Twelve stable major QTLs accounting for over 10 % of phenotypic variation were included in five optimal clusters on the genetic region of bins 1.02-1.03, 1.04-1.06, 2.05-2.07, 4.07-4.08 and 9.03-9.04; the addition and partial dominance effects of significant QTL play an important role in controlling the development of maize kernel. These putative QTL may have great promising for further fine-mapping with more markers, and genetic improvement of maize kernel size and weight through marker-assisted breeding. [ABSTRACT FROM AUTHOR]

The lepidopteran Ostrinia furnacalis is one of the most serious pests of maize production. The Cry1C proteins are group of Bacillus thuringiensis ( Bt) proteins that are toxic to the intestine of insects. Overexpression of Cry1C protein has led to increased resistance to lepidopteran pests in several crops. In the present study, the synthetic cry1C* gene that was previously tested in rice was introduced into maize Hi-II genotypes via biolistic gun-mediated transformation. A total of nine independent putative callus were obtained and 87 transgenic plants were positive with cry1C* according to polymerase chain reaction (PCR) analysis. Three highly insect-resistant transgenic plants, ZmKc-2-3, ZmKc-3-2, and ZmKc-3-5, were further confirmed by PCR analysis, field assessment, and genomic southern blotting in the T generations. Insect bioassays were conducted in both the field and the laboratory, and showed that progeny of the three transgenic lines were significantly resistant to lepidopteran maize pests during the whole development and growth period. The stable integration and expression of the cry1C* in the three transgenic plants' progeny were confirmed by reverse transcription-PCR (RT-PCR) and enzyme-linked immuno sorbent assay (ELISA) methods. Hybrids were produced by crossing transgenic line ZmKc-2-3 with the elite inbred line Zheng58. There was small variation among the hybrids and backcross offspring, indicating that these cry1C* transgenic lines can be used to produce insect-resistant hybrids and served as insect-resistant sources for the development of Bt maize. [ABSTRACT FROM AUTHOR]

Drought can cause severe reduction in maize production, and strongly threatens crop yields. To dissect this complex trait and identify superior alleles, 350 tropical and subtropical maize inbred lines were genotyped using a 1536-SNP array developed from drought-related genes and an array of 56,110 random SNPs. The inbred lines were crossed with a common tester, CML312, and the testcrosses were phenotyped for nine traits under well-watered and water-stressed conditions in seven environments. Using genome-wide association mapping with correction for population structure, 42 associated SNPs ( P ≤ 2.25 × 10 0.1/ N) were identified, located in 33 genes for 126 trait × environment × treatment combinations. Of these genes, three were co-localized to drought-related QTL regions. Gene GRMZM2G125777 was strongly associated with ear relative position, hundred kernel weight and timing of male and female flowering, and encodes NAC domain-containing protein 2, a transcription factor expressed in different tissues. These results provide some good information for understanding the genetic basis for drought tolerance and further studies on identified candidate genes should illuminate mechanisms of drought tolerance and provide tools for designing drought-tolerant maize cultivars tailored to different environmental scenarios. [ABSTRACT FROM AUTHOR]

To investigate the genetic basis of maize seedling response to waterlogging, we performed a genome-wide association study in 144 maize inbred lines, measuring length, fresh and dry weight of roots and shoots under normal and waterlogged conditions using 45,868 SNPs. This panel was divided into three subgroups based on the population structure results and the LD decay distance was 180 kb. A biparental advanced backcross (AB) population was also used to detect quantitative trait loci (QTL). In a comparison of 16 different models, principal components analysis (PCA/top PC3) + K was found to be best for reduction of false-positive associations for further analysis. A whole-genome scan detected four strong peak signals ( P < 2.18 × 10) significantly associated with the waterlogging response on chromosomes 5, 6 and 9. SNP4784, SNP200, SNP298, and SNP6314 showed significant association with corresponding traits under waterlogging and explained 14.99-19.36 %, 15.75-17.64 %, 16.08 % and 15.44 % of the phenotypic variation, respectively. The identified SNPs were located in GRMZM2G012046, GRMZM2G009808, GRMZM2G137108 and GRMZM2G369629 (AGPV1). SNP4784 (GRMZM2G012046) was colocalized with the major QTL that was identified with the same traits in the AB population. Forty-seven SNPs significantly associated ( P < 2.18 × 10) with six traits in association mapping were identified and, among these, 33 SNPs were already reported in literature as waterlogging-related traits. These results will help elucidate the genetic basis of differential responses and tolerance to waterlogging stress among maize inbred lines, and provide novel loci for improvement of waterlogging tolerance of maize inbred lines using marker-assisted selection. [ABSTRACT FROM AUTHOR]

Sporisorium reilianum f. sp. zeae is an important biotrophic pathogen that causes head smut disease in maize. Head smut is not obvious until the tassels and ears emerge. S. reilianum has a very long life cycle that spans almost the entire developmental program of maize after the pathogen successfully invades the root. The aim of this study was to understand at a molecular level how this pathogen interacts with the host during its long life cycle, and how this interaction differs between susceptible and resistant varieties of maize after hyphal invasion. We investigated transcriptional changes in the resistant maize line Mo17 at four developmental stages using a maize 70mer-oligonucleotide microarray. We found that there was a lengthy compatible relationship between the pathogen and host until the early eighth-leaf stage. The resistance in Mo17 relied on the assignment of auxin and regulation of flavonoids in the early floral primordium during the early floral transition stage. We propose a model describing the putative mechanism of head smut resistance in Mo17 during floral transition. In the model, the synergistic regulations among auxin, flavonoids, and hyphal growth play a key role in maintaining compatibility with S. reilianum in the resistant maize line. [ABSTRACT FROM AUTHOR]

The maize smut fungus, Sporisorium reilianum f. sp. zeae, which is an important biotrophic pathogen responsible for extensive crop losses, can infect maize by invading the root during the early seedling stage. In order to investigate disease-resistance mechanisms at this early seedling stage, digital gene expression analysis, which applies a dual-enzyme approach, was used to identify the transcriptional changes in the roots of Huangzao4 (susceptible) and Mo17 (resistant) after root inoculation with S. reilianum. During the infection in the roots, the expression pattern of pathogenesis-related genes in Huangzao4 and Mo17 were significantly differentially regulated at different infection stages. The glutathione S-transferase enzyme activity and reactive oxygen species levels also showed changes before and after inoculation. The total lignin contents and the pattern of lignin depositions in the roots differed during root colonization of Huangzao4 and Mo17. These results suggest that the interplay between S. reilianum and maize during the early infection stage involves many important transcriptional and physiological changes, which offer several novel insights to understanding the mechanisms of resistance to the infection of biotrophic fungal pathogens. [ABSTRACT FROM AUTHOR]

Combining ability is essential for hybrid breeding in crops. However, the genetic basis of combining ability remains unclear and has been seldom investigated. Identifying molecular markers associated with this complex trait would help to understand its genetic basis and provide useful information for hybrid breeding in maize. In this study, we identified genetic loci of general combining ability (GCA) and specific combining ability (SCA) for five yield-related traits under three environments using a set of testcrosses with introgression lines (ILs). GCA or SCA of the five yield-related traits of the ILs was estimated by the performance of testcrosses with four testers from different heterotic groups. Genetic correlations between GCA of the traits and the corresponding traits per se were not significant or not strong, suggesting that the genetic basis between them is different. A total of 56 significant loci for GCA and 21 loci for SCA were commonly identified in at least two environments, and only 5 loci were simultaneously controlling GCA and SCA, indicating that the genetic basis of GCA and SCA is different. For all of the traits investigated, positive and significant correlations between the number of GCA loci in the ILs and the performance of the corresponding GCA of the ILs were detected, implying that pyramiding GCA loci would have positive effect on the performance of GCA. Results in this study would be useful for maize hybrid breeding. [ABSTRACT FROM AUTHOR]

Maize ( Zea mays L.), one of the most important cereal crops in the world, is salt-sensitive. It is critical to study the genes associated with salt tolerance, the differential gene expression pattern between salt-tolerant and salt-sensitive lines, and the mechanism of salt tolerance in maize. Two maize inbred lines with different salt sensitivity, NC286 and Huangzao4, were used to reveal early responsive genes under high salinity condition. Suppression subtractive hybridization (SSH) assay was conducted to identify potential genes involved in the early stage of maize seedlings in responses to 200mM NaCl. A custom μparaflo™ microfluidic array (LC science USA) containing Release version 10.1 plant miRNA probes () was used to discover salt stress responsive miRNAs using the differences in miRNAs expression between these two maize lines. Cis-elemnet analysis was performed in the promoter regions of these two kinds of salt stress responsive transcriptions. The expression of a total of 141 unique genes was significantly changed under 200mM NaCl salt shock within 24 hours. These genes were involved in a broad spectrum of biochemical, cellular, and physiological processes. They were assigned to 14 categories based on their biological functions. Some of them were previously reported to be associated with biotic and abiotic stresses, and several novel genes detected. A gene model was proposed which showed how these genes could sense and response to the high salinity environment. The promoter regions of these 141 salt responsive genes were analyzed, and salt responsive key cis-elements uncovered. The cis-elements usage bias of the salt responsive mRNAs and non-coding RNAs were of the same. The different expression profiles of the genes, especially the ones involved in salt signal cascades and phytohormones responding, could explain the different salt stress response in the two maize inbred lines. The similar cis-elements usage bias of two kinds of salt responsive transcriptions suggests they may be regulated by the same trans-factors. [ABSTRACT FROM AUTHOR]

Breeding maize varieties for high nitrogen (N) use efficiency (NUE) by marker-assisted selection using NUE quantitative trait locus (QTL) or by genetic transfer of NUE-associated genes is a viable approach for increasing grain yield in N-limited production areas. In this investigation, we evaluated a set of introgression line populations under N supply and N deficiency conditions. From 42 QTLs for grain yield and yield components, 23 were identified under N supply conditions and 33 from N limited conditions. Meta-analysis of published maize NUE QTLs revealed 37 'consensus' QTLs, of which, 18 was detected under low N conditions. In addition, 258 unique ESTs associated with low N stress response, N uptake, transport, and assimilation were aligned on the maize genome by in silico mapping. Integrating the EST map with the QTL map has resulted in the identification of candidate NUE-associated genes of the following functional categories: N uptake, transport, and assimilation; carbon (C) metabolism and assimilation; and cascades of stress response and signal transduction genes. Nine candidates that have been introgressed into Ye478 significantly altered grain yield/yield components. It is suggested that the dynamics of interactions between C and N metabolism are important for maize yield. A high NUE variety should have a highly efficient C assimilation per unit N and actively express CO assimilation-related genes under N-limited conditions. [ABSTRACT FROM AUTHOR]

The rice BAC-DNA was used as probes and fluorescence in situ hybridization (FISH) was applied to the interphase and metaphase mitotic chromosomes of maize. To optimize the BAC-FISH technique, we respectively assayed the effect of several factors, including maize or rice genomic C o t DNA used as blocking reagent of DNA, washing temperatures and FAD concentration in the washing buffer and in the hybrid solution. The results show that C o t DNA of maize genome blocked the repetitive sequence of the rice BAC-DNA when the C o t value was below 50. Meanwhile, it was necessary to adjust the C o t value according to the different probes and their ratios. Decreasing the concentration of FAD in the hybridization mixtures, adjusting the washing rate after hybridization, and most especially, blocking the ricespecific repetitive sequences of BAC-DNA could improve the positive signals of BAC-FISH. [ABSTRACT FROM AUTHOR]

A total of 26718 M1 plants were obtained by crossing the active mutator transposon donor parents (Q105, WW51, 115F, V26-2 and 919J) with the recipient parents (Hz85,W328 with Bz gene and S-Mo17R13R13). The phenotypes of M1 plants were observed in the field. M1 plants were self-pollinated to develop the mutator insertion-mutagenized M2 seeds. The transposition frequency of the mutator in the genome was calculated based on the spotted aleurone phenotype of the M2 seeds. The results showed that: (1) the mutation frequency of M1 phenotypes in the field was 0.07 in the population of W328xMu; (2) the mutation frequency of spotted aleurone seeds on the M2 ears was 0.122 in the population of W328xMu; (3) five S-cytoplasm male-sterile plants were found among 22500 M1 plants of S-Mo17R13R13xMu, with the transposition frequency about 2.2x10-4 per locus. 99 flanking sequences of mutator transposition were amplified by the modified MuTAIL-PCR, and 59 non-redundant sequences with length around 400 bp were obtained. After bioinformatic analysis, 27 sequences of them could be annotated, using non-redundant nucleotide database of maize, rice, and Arabidopsis. 36 sequences of them were located on the genetic map of maize by comparative genomics, and several flanking sequences of mutator insertion were mapped on the single marker locus. Hotspot sequences of mutator transposition were revealed by comparing the homologies between the 9-bp target site duplication of the mutator insertion. The putative functions of 8 flanking sequences of mutator transposition had identity with the functions of their corresponding marker. The constructed mutator insertion mutant population in maize will facilitate the new gene discovery and functional genomics study in maize. [ABSTRACT FROM AUTHOR]

1201 published maize QTLs conferring for 68 traits were collected and imported into local CMap software to construct an integrated QTL map, which can be used for marker-mining, QTL localization, gene cloning and marker-assisted selection. The maize integrated QTL map showed that maize QTLs for various traits usually clustered in all chromosomes. 22 plant height QTLs of maize were co-linear with 64 plant height QTLs of rice, 43 grain yield QTLs of maize were co-liner with 7 grain yield QTLs of rice. 127 plant height QTLs of maize were refined by means of "overview" analysis. At last, 40 "real" QTLs were identifed. A substantial number of candidate quantitative trait genes for plant height of maize were found. These results established an important bioinformatics platform for extracting most of maize QTL information. [ABSTRACT FROM AUTHOR]

By adding thirty-one markers in the previous linkage map, a new genetic linkage map containing 205 markers was constructed, spanning a total of 2305.4 cM with an average interval of 11.2 cM. The genotypic errors in the whole genome were detected by the statistical method and removed manually. The precision of the linkage map was improved significantly. Main and epistatic QTL were detected by R/qtl, and main QTL were confirmed and refined by multiple interval mapping (MIM). Finally, MIM detected seven QTL for rows number, and five QTL for each grain yield, kernels per row and 100-kernel weight. The contribution to genetic variations of QTL varied from 35.3% for grain yield to 61.5% for rows number. Only kernels per row exhibited significant epistatic interactions between QTL. Twenty-four epistatic QTL were detected which distributed on almost all the ten chromosomes. About two-third epistatic QTL were observed between main QTL and another locus, which had no significant effects. These results indicate rather clearly that there are a number of QTL affecting trait expressions, not directly but indirectly through interactions with other loci. Thus, epistatic QTL effects may play a crucial role, if not more important than main QTL effects, in the genetic variation for the measured traits in present study. [ABSTRACT FROM AUTHOR]

In this study, SSH (Suppression Subtractive Hybridization) and cDNA microarray were used to identify genes associated with waterlogging response of maize roots. Mo17 and Hz32 are two maize inbred lines with differential tolerance to hypoxia. Seedlings of the inbred lines with two leaves were submerged in hypoxia buffer. SSH libraries were constructed with cDNA samples from roots. Both forward and reverse subtractions were performed for each inbred line, and 105 positive clones induced by hypoxia were selected by differential screening. The treated and control message RNA were hybridized with the cDNA microarray of Mo17, sequentially, 57 of 3-fold differentially expressed clones were obtained. A total of 162 positive clones were all sequenced. Bioinformatics analysis showed these positive clones represent 85 TUGs, including genes involved in several biochemistry pathways, such as glycolysis, protection, signal transduction, cell construction and energy metabolism and 41 EST with unknown function. Comparison between Mo17 and Hz32 indicates that genes related to hypoxia tolerance have different expression patterns in submerged roots. Several positive clones’ expression patterns were revealed by Northern or RT-PCR, and a new gene (Sicyp51), which may contribute to hypoxia tolerance, was identified. [ABSTRACT FROM AUTHOR]

Studies plant height in maize, an important agronomic and model trait that is suitable for studying developmental biology. Use of data from locations in Wuhan and Xiangfan and a molecular linkage map covering 10 chromosomes; Analysis of QTLs affecting plant height at five different developmental stages by the combination of composite interval mapping and the conditional analysis method; Number of QTLs located at different regions of five chromosomes; Finding that there were different QTL effect values on plant height at various developmental stages; Indication that three QTLs were detected at all five stages; Suggestion that QTL expression varied at different stages; Detection of seven conditional QTLs for plant height; Indication that QTLs for plant height are expressed in different time-space; Implication for marker assistant selection when conducted for quantitative traits.

Presents a study which developed a method to construct S-type cytoplasm male sterility (CMS) maize mitochondrial gene library by means of embedding mitochondria and enzymatic digesting mitochondria in situ. Information on CMS; Materials and method; Results and discussion.

A correction to the article "Digital gene expression analysis of early root infection resistance to Sporisorium reilianum f. sp. zeae in maize" that was published in the January 2013 issue is presented.