Genome-Wide Identification and Expression Profile Reveal Potential Roles of Peanut <i>ZIP</i> Family Genes in Zinc/Iron-Deficiency Tolerance

Zinc/iron-regulated transporter-like protein (ZIP) family genes play crucial roles in metal uptake and transport in plants. However, little is known about their functions in peanut. Here, genome-wide analysis identified 30 peanut AhZIP genes that were divided into four classes. Most AhZIPs experienced whole-genome or segmental duplication. AhZIP proteins harbored 3–8 transmembrane domains and a typical ZIP domain, showing considerable homology with BbZIP from Bordetella bronchiseptica. Clustered AhZIPs generally share similar gene/protein structures; however, unique features were found in AhIRT1.2, AhZIP1.2, AhZIP3.5 and AhZIP7.8. RNA-seq data revealed that AhZIP2.1/2.2, AhZIP4.1/4.2 and AhZIP11.1/11.2 were highly and preferentially expressed in roots, nodule and reproductive tissues. RT-qPCR analysis indicated that transcriptional responses of AhZIPs to Fe/Zn deficiency are cultivar dependent. The expressions of AhIRT1.1, AhIRT1.2 and AhZIP6.1 were closely related to Fe uptake and translocation. AhIRT1.1 and AhZIP7.2 expression were significantly correlated with Zn accumulation. The expression of AhIRT1.1, AhIRT1.2, AhZIP3.6, AhZIP6.1 and AhZIP11.1 was associated with Mn uptake and translocation. The results confirmed that AhZIP genes play crucial roles in the uptake and transport of Fe, Zn and Mn in peanut, providing clues to further functionally characterize AhZIP genes in the future

Comparative proteomics analysis of peanut roots reveals differential mechanisms of cadmium detoxification and translocation between two cultivars differing in cadmium accumulation

Abstract Background Peanut is one of the most important oil and protein crops, and it exhibits wide cultivar variations in shoot Cd accumulation ability. However, the mechanism of Cd accumulation in peanut shoots has not been well understood. In this study, the root proteomics of two cultivars differing in seed Cd accumulation, Fenghua 1 (F, low Cd cultivar) and Silihong (S, high Cd cultivar), were investigated under 0 (CK) and 2 μM Cd conditions. Results A total of 4676 proteins were identified by proteomics screening. Of them, 375, 1762, 1276 and 771 proteins were identified to be differentially expressed proteins (DEPs) for comparison of FCd/FCK, SCd/SCK, FCK/SCK and FCd/SCd, respectively. Silihong is more sensitive to Cd exposure than Fenghua 1 in terms of root proteomics. A total of 30 and 86 DEPs were identified to be related with heavy metal transport and cell wall modification, respectively. The up-regulation of ABCB25, ABCC14, ABCC2, PDR1 and V-ATPases by Cd exposure in Silihong might enhance vacuolar sequestration of Cd and its efflux from symplast to apoplast. The higher Cd accumulation in the root CWs of Silihong might be resulted from its higher capability of CW modification, in which many proteins such as IRX10L, BGLU12-like, BGLU42, EXLB1, XTH30, XTH6, XYL7, PAL3, COMT, CAD1, and CCR1 were involved. Conclusions The vacuolar sequestration and efflux of Cd as well as its adsorption in CW might be the principal mechanism of cadmium detoxification in Silihong. The higher capacity of Cd accumulation and translocation of Silihong is an inherent characteristics in which ACA8 and ZIP1 might be involved

Comparative transcriptome analysis reveals gene network regulating cadmium uptake and translocation in peanut roots under iron deficiency

Abstract Background Iron (Fe) is an essential element for plant growth and development, whereas cadmium (Cd) is non-essential and highly toxic. Previous studies showed that Fe deficiency enhanced Cd uptake and accumulation in peanuts. However, the molecular mechanism underlying the increased Cd accumulation in Fe-deficient peanut plants is poorly understood. Results We employed a comparative transcriptome analysis approach to identify differentially expressed genes (DEGs) in peanut roots exposed to Fe-sufficient without Cd, Fe-deficient without Cd, Fe-sufficient with Cd and Fe-deficient with Cd. Compared with the control, Fe deficiency induced 465 up-regulated and 211 down-regulated DEGs, whereas the up- and down-regulated DEGs in Cd exposed plants were 329 and 189, respectively. Under Fe-deficient conditions, Cd exposure resulted in 907 up-regulated DEGs and 953 down-regulated DEGs. In the presence of Cd, Fe deficiency induced 1042 up-regulated and 847 down-regulated genes, respectively. Based on our array data, we found that metal transporter genes such as CAX4, COPT1, IRT1, NRAMP5, OPT3, YSL3, VIT3 and VIT4 might be involved in iron homeostasis. Moreover, combined with quantitative real-time PCR, IRT1, NRAMP3, NRAMP5, OPT3, YSL3, ABCC3, ZIP1, and ZIP5 were verified to be responsible for Cd uptake and translocation in peanut plants under iron deficiency. Additionally, a larger amount of ABC transporter genes was induced or suppressed by iron deficiency under Cd exposure, indicating that this family may play important roles in Fe/Cd uptake and transport. Conclusions The up-regulated expression of NRAMP5 and IRT1 genes induced by iron deficiency may enhance Cd uptake in peanut roots. The decrease of Cd translocation from roots to shoots may be resulted from the down-regulation of ZIP1, ZIP5 and YSL3 under iron deficiency

Comparative transcriptome analysis reveals key cadmium transport-related genes in roots of two pak choi (Brassica rapa L. ssp. chinensis) cultivars

Abstract Background Cadmium translocation from roots to shoots is a complex biological process that is controlled by gene regulatory networks. Pak choi exhibits wide cultivar variations in Cd accumulation. However, the molecular mechanism involved in cadmium translocation and accumulation is still unclear. To isolate differentially expressed genes (DEGs) involved in transporter-mediated regulatory mechanisms of Cd translocation in two contrasting pak choi cultivars, Baiyewuyueman (B, high Cd accumulator) and Kuishan’aijiaoheiye (K, low Cd accumulator), eight cDNA libraries from the roots of two cultivars were constructed and sequenced by RNA-sequencing. Results A total of 244,190 unigenes were obtained. Of them, 6827 DEGs, including BCd10 vs. BCd0 (690), KCd10 vs. KCd0 (2733), KCd0 vs. BCd0 (2919), and KCd10 vs. BCd10 (3455), were identified. Regulatory roles of these DEGs were annotated and clarified through GO and KEEG enrichment analysis. Interestingly, 135 DEGs encoding ion transport (i.e. ZIPs, P1B-type ATPase and MTPs) related proteins were identified. The expression patterns of ten critical genes were validated using RT-qPCR analysis. Furthermore, a putative model of cadmium translocation regulatory network in pak choi was proposed. Conclusions High Cd cultivar (Baiyewuyueman) showed higher expression levels in plasma membrane-localized transport genes (i.e., ZIP2, ZIP3, IRT1, HMA2 and HMA4) and tonoplast-localized transport genes (i.e., CAX4, HMA3, MRP7, MTP3 and COPT5) than low Cd cultivar (Kuishan’aijiaoheiye). These genes, therefore, might be involved in root-to-shoot Cd translocation in pak choi

Genome-Wide Identification and Expression Analysis Reveals Roles of the <i>NRAMP</i> Gene Family in Iron/Cadmium Interactions in Peanut

The natural resistance-associated macrophage protein (NRAMP) family plays crucial roles in metal uptake and transport in plants. However, little is known about their functions in peanut. To understand the roles of AhNRAMP genes in iron/cadmium interactions in peanut, genome-wide identification and bioinformatics analysis was performed. A total of 15 AhNRAMP genes were identified from the peanut genome, including seven gene pairs derived from whole-genome duplication and a segmental duplicated gene. AhNRAMP proteins were divided into two distinct subfamilies. Subfamily I contains eight acid proteins with a specific conserved motif 7, which were predicted to localize in the vacuole membrane, while subfamily II includes seven basic proteins sharing specific conserved motif 10, which were localized to the plasma membrane. Subfamily I genes contained four exons, while subfamily II had 13 exons. AhNRAMP proteins are perfectly modeled on the 5m94.1.A template, suggesting a role in metal transport. Most AhNRAMP genes are preferentially expressed in roots, stamens, or developing seeds. In roots, the expression of most AhNRAMPs is induced by iron deficiency and positively correlated with cadmium accumulation, indicating crucial roles in iron/cadmium interactions. The findings provide essential information to understand the functions of AhNRAMPs in the iron/cadmium interactions in peanuts

Comparative transcriptome analysis revealed key factors for differential cadmium transport and retention in roots of two contrasting peanut cultivars

Abstract Background Peanut is the world’s fourth largest oilseed crop that exhibits wide cultivar variations in cadmium (Cd) accumulation. To establish the mechanisms of Cd distribution and accumulation in peanut plants, eight cDNA libraries from the roots of two contrasting cultivars, Fenghua 1 (low-Cd cultivar) and Silihong (high-Cd cultivar), were constructed and sequenced by RNA-sequencing. The expression patterns of 16 candidate DEGs were validated by RT-qPCR analysis. Results A total of 75,634 genes including 71,349 known genes and 4484 novel genes were identified in eight cDNA libraries, among which 6798 genes were found to be Cd-responsive DEGs and/or DEGs between these two cultivars. Interestingly, 183 DEGs encoding ion transport related proteins and 260 DEGs encoding cell wall related proteins were identified. Among these DEGs, nine metal transporter genes (PDR1, ABCC4 and ABCC15, IRT1, ZIP1, ZIP11, YSL7, DTX43 and MTP4) and nine cell wall related genes (PEs, PGIPs, GTs, XYT12 CYP450s, LACs, 4CL2, C4H and CASP5) showed higher expression in Fenghua 1 than in Silihong. Conclusions Both the metal transporters and cell wall modification might be responsible for the difference in Cd accumulation and translocation between Fenghua 1 and Silihong. These findings would be useful for further functional analysis, and reveal the molecular mechanism responsible for genotype difference in Cd accumulation

AhIRT1 and AhNRAMP1 metal transporter expression correlates with Cd uptake in peanuts under iron deficiency - Fig 4

<p>Expression pattern of <i>AhNRAMP1</i> (a) and <i>AhIRT1</i> (b) in the roots of Luhua 8 and Zhenghong 3 grown in full nutrient solution (+Fe) or without Fe (−Fe) for 12 d. The expression levels of <i>AhNRAMP1</i> and <i>AhIRT1</i> were normalized to that of <i>Ahactin</i> gene. Different letters above error bars indicate values (mean ± SE, n = 3) are significantly different between treatments at the 0.05 level.</p

Growth of wild-type yeast cells transformed with empty vector pYES2, <i>AhIRT1</i> and <i>AhNRAMP1</i> in the presence of galactose.

<p>Serial dilutions of the transformed yeast cells with OD_600nm 0.5 to 0.0005 were spotted on SD-Ura plates containing 0 or 30 μM CdCl₂ in the presence of galactose. The yeast was grown on the plates at 30°C for 3 d for the comparison.</p

Parameters of the Michaelis-Menten plus linear model used to resolve the kinetic of influx curves in Fig 3.

<p>Data represent mean values ± SE.</p