Jingtang Chen

Identification and characterization of the zinc-regulated transporters, iron-regulated transporter-like protein (ZIP) gene family in maize

BackgroundZinc (Zn) and iron (Fe) are essential micronutrients for plant growth and development, their deficiency or excess severely impaired physiological and biochemical reactions of plants. Therefore, a tightly controlled zinc and iron uptake and homeostasis network has been evolved in plants. The Zinc-regulated transporters, Iron-regulated transporter-like Proteins (ZIP) are capable of uptaking and transporting divalent metal ion and are suggested to play critical roles in balancing metal uptake and homeostasis, though a detailed analysis of ZIP gene family in maize is still lacking.ResultsNine ZIP-coding genes were identified in maize genome. It was revealed that the ZmZIP proteins share a conserved transmembrane domain and a variable region between TM-3 and TM-4. Transiently expression in onion epidermal cells revealed that all ZmZIP proteins were localized to the endoplasmic reticulum and plasma membrane. The yeast complementation analysis was performed to test the Zn or Fe transporter activity of ZmZIP proteins. Expression analysis showed that the ZmIRT1 transcripts were dramatically induced in response to Zn- and Fe-deficiency, though the expression profiles of other ZmZIP changed variously. The expression patterns of ZmZIP genes were observed in different stages of embryo and endosperm development. The accumulations of ZmIRT1 and ZmZIP6 were increased in the late developmental stages of embryo, while ZmZIP4 was up-regulated during the early development of embryo. In addition, the expression of ZmZIP5 was dramatically induced associated with middle stage development of embryo and endosperm.ConclusionsThese results suggest that ZmZIP genes encode functional Zn or Fe transporters that may be responsible for the uptake, translocation, detoxification and storage of divalent metal ion in plant cells. The various expression patterns of ZmZIP genes in embryo and endosperm indicates that they may be essential for ion translocation and storage during differential stages of embryo and endosperm development. The present study provides new insights into the evolutionary relationship and putative functional divergence of the ZmZIP gene family during the growth and development of maize.

The genetic architecture of zinc and iron content in maize grains as revealed by QTL mapping and meta-analysis.

Micronutrient malnutrition, especially zinc (Zn) and iron (Fe) deficiency in diets, has aroused worldwide attention. Biofortification of food crops has been considered as a promising approach for alleviating this deficiency. Quantitative trait locus (QTL) analysis was performed to dissect the genetic mechanism of Zn and Fe content in maize grains using a total of 218 F2:3 families derived from a cross between inbred lines 178 and P53. Meta-analysis was used to integrate genetic maps and detect Meta-QTL (MQTL) across several independent QTL researches for traits related to Zn or Fe content. Five significant QTLs and 10 MQTLs were detected. Two informative genomic regions, bins 2.07 and 2.08, showed a great importance for Zn and Fe content QTLs. The correlation between Zn and Fe level in maize grains was proposed by MQTLs as 8 of the 10 involved both traits. The results of this study suggest that QTL mapping and meta-analysis is an effective approach to understand the genetic basis of Zn and Fe accumulation in maize grains.

Overexpression of ZmIRT1 and ZmZIP3 Enhances Iron and Zinc Accumulation in Transgenic Arabidopsis

Iron and zinc are important micronutrients for both the growth and nutrient availability of crop plants, and their absorption is tightly controlled by a metal uptake system. Zinc-regulated transporters, iron-regulated transporter-like proteins (ZIP), is considered an essential metal transporter for the acquisition of Fe and Zn in graminaceous plants. Several ZIPs have been identified in maize, although their physiological function remains unclear. In this report, ZmIRT1 was shown to be specifically expressed in silk and embryo, whereas ZmZIP3 was a leaf-specific gene. Both ZmIRT1 and ZmZIP3 were shown to be localized to the plasma membrane and endoplasmic reticulum. In addition, transgenic Arabidopsis plants overexpressing ZmIRT1 or ZmZIP3 were generated, and the metal contents in various tissues of transgenic and wild-type plants were examined based on ICP-OES and Zinpyr-1 staining. The Fe and Zn concentration increased in roots and seeds of ZmIRT1-overexpressing plants, while the Fe content in shoots decreased. Overexpressing ZmZIP3 enhanced Zn accumulation in the roots of transgenic plants, while that in shoots was repressed. In addition, the transgenic plants showed altered tolerance to various Fe and Zn conditions compared with wild-type plants. Furthermore, the genes associated with metal uptake were stimulated in ZmIRT1 transgenic plants, while those involved in intra- and inter- cellular translocation were suppressed. In conclusion, ZmIRT1 and ZmZIP3 are functional metal transporters with different ion selectivities. Ectopic overexpression of ZmIRT1 may stimulate endogenous Fe uptake mechanisms, which may facilitate metal uptake and homeostasis. Our results increase our understanding of the functions of ZIP family transporters in maize.

Comparative mapping combined with homology-based cloning of the rice genome reveals candidate genes for grain zinc and iron concentration in maize

BackgroundGrain zinc and iron concentration is a complex trait that is controlled by quantitative trait loci (QTL) and is important for maintaining body health. Despite the substantial effort that has been put into identifying QTL for grain zinc and iron concentration, the integration of independent QTL is useful for understanding the genetic foundation of traits. The number of QTL for grain zinc and iron concentration is relatively low in a single species. Therefore, combined analysis of different genomes may help overcome this challenge.ResultsAs a continuation of our work on maize, meta-analysis of QTL for grain zinc and iron concentration in rice was performed to identify meta-QTL (MQTL). Based on MQTL in rice and maize, comparative mapping combined with homology-based cloning was performed to identify candidate genes for grain zinc and iron concentration in maize. In total, 22 MQTL in rice, 4 syntenic MQTL-related regions, and 3 MQTL-containing candidate genes in maize (ortho-mMQTL) were detected. Two maize orthologs of rice, GRMZM2G366919 and GRMZM2G178190, were characterized as natural resistance-associated macrophage protein (NRAMP) genes and considered to be candidate genes. Phylogenetic analysis of NRAMP genes among maize, rice, and Arabidopsis thaliana further demonstrated that they are likely responsible for the natural variation of maize grain zinc and iron concentration.ConclusionsSyntenic MQTL-related regions and ortho-mMQTL are prime areas for future investigation as well as for marker-assisted selection breeding programs. Furthermore, the combined method using the rice genome that was used in this study can shed light on other species and help direct future quantitative trait research. In conclusion, these results help elucidate the molecular mechanism that underlies grain zinc and iron concentration in maize.

QTL Mapping for Stalk Related Traits in Maize (Zea mays L.) Under Different Densities

Abstract Stalk related traits, comprising plant height (PH), ear height (EH), internode number (IN), average internode length (AIL), stalk diameter (SD), and ear height coefficient (EHC), are significantly correlated with yield, density tolerance, and lodging resistance in maize. To investigate the genetic basis for stalk related traits, a doubled haploid (DH) population derived from a cross between NX531 and NX110 were evauluated under two densities over 2 yr. The additive quantitative trait loci (QTLs), epistatic QTLs were detected using inclusive composite interval mapping and QTL-by-environment interaction were detected using mixed linear model. Differences between the two densities were significant for the six traits in the DH population. A linkage map that covered 1 721.19 cM with an average interval of 10.50 cM was constructed with 164 simple sequence repeat (SSR). Two, two, seven, six, two, and eight additive QTLs for PH, IN, AIL, EH, SD, and EHC, respectively. The extend of their contribution to penotypic variation ranged from 10.10 to 31.93%. Seven QTLs were indentified simultaneously under both densities. One pair, two pairs and one pair of epistatic effects were detected for AIL, SD and EHC, respectively. No epistatic effects were detected for PH, EH, and IN. Nineteen QTLs with environment interactions were detected and their contribution to phenotypic variation ranged from 0.43 to 1.89%. Some QTLs were stably detected under different environments or genetic backgrounds comparing with previous studies. These QTLs could be useful for genetic improvement of stalk related traits in maize breeding.

Is there a strategy I iron uptake mechanism in maize

ABSTRACT Iron is a metal micronutrient that is essential for plant growth and development. Graminaceous and nongraminaceous plants have evolved different mechanisms to mediate Fe uptake. Generally, strategy I is used by nongraminaceous plants like Arabidopsis, while graminaceous plants, such as rice, barley, and maize, are considered to use strategy II Fe uptake. Upon the functional characterization of OsIRT1 and OsIRT2 in rice, it was suggested that rice, as an exceptional graminaceous plant, utilizes both strategy I and strategy II Fe uptake systems. Similarly, ZmIRT1 and ZmZIP3 were identified as functional zinc and iron transporters in the maize genome, along with the determination of several genes encoding Zn and Fe transporters, raising the possibility that strategy I Fe uptake also occurs in maize. This mini-review integrates previous reports and recent evidence to obtain a better understanding of the mechanisms of Fe uptake in maize.

Constitutive expression of the ZmZIP7 in Arabidopsis alters metal homeostasis and increases Fe and Zn content.

Iron (Fe) and zinc (Zn) are important micronutrients for plant growth and development. Zinc-regulated transporters and the iron-regulated transporter-like protein (ZIP) are necessary for the homeostatic regulation of these metal micronutrients. In this study, the physiological function of ZmZIP7 which encodes a ZIP family transporter was characterized. We detected the expression profiles of ZmZIP7 in maize, and found that the accumulation of ZmZIP7 in root, stem, leaf, and seed was relatively higher than tassel and young ear. ZmZIP7 overexpression transgenic Arabidopsis lines were generated and the metal contents in transgenic and wild-type (WT) plants were examined using inductively coupled plasma atomic emission spectroscopy (ICP-OES) and Zinpyr-1 staining. Fe and Zn concentrations were elevated in the roots and shoots of ZmZIP7-overexpressing plants, while only Fe content was elevated in the seeds. We also analyzed the expression profiles of endogenous genes associated with metal homeostasis. Both endogenic Fe-deficiency inducible genes and the genes responsible for Zn and Fe transport and storage were stimulated in ZmZIP7 transgenic plants. In conclusion, ZmZIP7 encodes a functional Zn and Fe transporter, and ectopic overexpression of ZmZIP7 in Arabidopsis stimulate endogenous Fe and Zn uptake mechanisms, thereby facilitating both metal uptake and homeostasis. Our results contribute to improved understanding of ZIP family transporter functions and suggest that ZmZIP7 could be used to enhance Fe levels in grains.

Diversity, Structure, and Marker-Trait Association Analysis of the Maize Recombinant Inbred Line Population

Association mapping has emerged as a new tool to elucidate complex quantitative trait loci in maize, but there are few reports about systematic association analysis for the specific SSR markers with agronomic traits of interest in China. We investigated the morphological and genetic diversity and population structure for 76 maize recombinant inbred lines, and then association analysis were further performed between 48 simple sequence repeat loci and 17 morphological traits, consisting of nine ear-related traits and eight other traits. The 48 SSR markers were screened out and further classified into two groups including a group of loci in regions harboring reported quantitative trait loci that affect ear shape and a group of markers distributing on the whole genome randomly. The result indicated that the population of recombinant inbred lines was structured, showing five subpopulations. Our association results revealed that there were 82, 59, and 40 significant associations detected by K-test, logistic regression, and both analysis, respectively. When the 17 traits were considered separately, the significant associations between Q-SSRs and E-traits were raised to 27.8%, whereas the other groups of combinations ranged between 2.3 and 6.3%. As the proportion of significant associations is higher among the Q-SSR subset of markers and the subset of traits related to ear shape than those for all of the other combinations, we conclude that this approach is valid for establishing true positive marker-trait relationships. Our results also demonstrated that association mapping could complement and enhance previous QTL information for marker-assisted selection.

Correction to: Identification of quantitative trait locus and prediction of candidate genes for grain mineral concentration in maize across multiple environments

By mistake this article had a note indicating that the article is part of the following topical collection: “This article is part of the Topical Collection on Plant Breeding: the Art of Bringing Science to Life. Highlights of the 20th EUCARPIA General Congress, Zurich, Switzerland, 29 August–1 September 2016 Edited by Roland Kölliker, Richard G. F. Visser, Achim Walter & Beat Boller” The article however is not part of this collection and should be seen as such by the reader.