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DNA microarray revealed and RNAi plants confirmed key genes conferring low Cd accumulation in barley grains.

Sun H, Chen ZH, Chen F, Xie L, Zhang G, Vincze E, Wu F - BMC Plant Biol. (2015)

Bottom Line: Quantitative RT-PCR confirmed our microarray data.Novel transporter genes such as HvZIP3 and HvZIP8 were identified as being associated with low-grain-Cd-accumulation.In addition to advancing academic knowledge, our findings may also result in potential economic benefits for molecular breeding of low Cd accumulating barley and other crops.

View Article: PubMed Central - PubMed

Affiliation: Department of Agronomy, College of Agriculture and Biotechnology, Zijingang Campus, Zhejiang University, Hangzhou, 310058, PR China. sunhongyan-8@163.com.

ABSTRACT

Background: Understanding the mechanism of low Cd accumulation in crops is crucial for sustainable safe food production in Cd-contaminated soils.

Results: Confocal microscopy, atomic absorption spectrometry, gas exchange and chlorophyll fluorescence analyses revealed a distinct difference in Cd accumulation and tolerance between the two contrasting barley genotypes: W6nk2 (a low-grain-Cd-accumulating and Cd-sensitive genotype) and Zhenong8 (a high-grain-Cd-accumulating and tolerant genotype). A DNA microarray analysis detected large-scale changes of gene expression in response to Cd stress with a substantial difference between the two genotypes. Cd stress led to higher expression of genes involved in transport, carbohydrate metabolism and signal transduction in the low-grain-Cd-accumulating genotype. Novel transporter genes such as zinc transporter genes were identified as being associated with low Cd accumulation. Quantitative RT-PCR confirmed our microarray data. Furthermore, suppression of the zinc transporter genes HvZIP3 and HvZIP8 by RNAi silencing showed increased Cd accumulation and reduced Zn and Mn concentrations in barley grains. Thus, HvZIP3 and HvZIP8 could be candidate genes related to low-grain-Cd-accumulation.

Conclusion: Novel transporter genes such as HvZIP3 and HvZIP8 were identified as being associated with low-grain-Cd-accumulation. In addition to advancing academic knowledge, our findings may also result in potential economic benefits for molecular breeding of low Cd accumulating barley and other crops.

No MeSH data available.


Related in: MedlinePlus

Relative expression level of ZIP in ZIP-RNAi lines compared to the Golden Promise parent. W-ZIP3, transgenic plants with HvZIP3 RNAi in W6nk2; W-ZIP8, transgenic HvZIP8 RNAi in W6nk2; Z-ZIP8, transgenic HvZIP8 RNAi of Zhenong8. a T0 lines; b T1 lines. * and ** indicates significant difference between transgenic lines and the parental line at P < 0.05 and P < 0.01, respectively
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Fig5: Relative expression level of ZIP in ZIP-RNAi lines compared to the Golden Promise parent. W-ZIP3, transgenic plants with HvZIP3 RNAi in W6nk2; W-ZIP8, transgenic HvZIP8 RNAi in W6nk2; Z-ZIP8, transgenic HvZIP8 RNAi of Zhenong8. a T0 lines; b T1 lines. * and ** indicates significant difference between transgenic lines and the parental line at P < 0.05 and P < 0.01, respectively

Mentions: The physiological distinctions observed in response to Cd stress (Figs. 1 and 2, Additional file 1: Figure S1, Additional file 2: Figure S2 and Additional file 3: Figure S3) between the two unique barley genotypes led to further exploitation of their differences at the transcriptome level. Overall, we found that compared to the control plants, the gene expression profiles the leaves of two genotypes were significantly altered after 15 days in 5 μM Cd. Cadmium stress induced differential expression of 812 genes. Among these genes, 382 (131) and 303 (106) were up-regulated (down-regulated) in W6nk2 and Zhenong8, respectively, with a fold change of at least 2.0 (P ≤ 0.05). These genes represented 2.3 and 1.8 % of the total genes in the two genotypes (data not shown). There were distinct differences in gene expression between the two genotypes (Additional file 4: Figure S4, Additional file 5: Figure S5 and Fig. 5; Table 1, Additional file 6: Table S1, Additional file 7: Table S2, Additional file 8: Table S3 and Additional file 9: Table S4), with 338 genes being up-regulated in W6nk2 (W-up) but unchanged in Zhenong8 (Z-NC), while another 13 genes were up-regulated in W6nk2 (W-up), but down-regulated in Zhenong8 (Z-down) (Additional file 4: Figure S4A).Table 1


DNA microarray revealed and RNAi plants confirmed key genes conferring low Cd accumulation in barley grains.

Sun H, Chen ZH, Chen F, Xie L, Zhang G, Vincze E, Wu F - BMC Plant Biol. (2015)

Relative expression level of ZIP in ZIP-RNAi lines compared to the Golden Promise parent. W-ZIP3, transgenic plants with HvZIP3 RNAi in W6nk2; W-ZIP8, transgenic HvZIP8 RNAi in W6nk2; Z-ZIP8, transgenic HvZIP8 RNAi of Zhenong8. a T0 lines; b T1 lines. * and ** indicates significant difference between transgenic lines and the parental line at P < 0.05 and P < 0.01, respectively
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4623906&req=5

Fig5: Relative expression level of ZIP in ZIP-RNAi lines compared to the Golden Promise parent. W-ZIP3, transgenic plants with HvZIP3 RNAi in W6nk2; W-ZIP8, transgenic HvZIP8 RNAi in W6nk2; Z-ZIP8, transgenic HvZIP8 RNAi of Zhenong8. a T0 lines; b T1 lines. * and ** indicates significant difference between transgenic lines and the parental line at P < 0.05 and P < 0.01, respectively
Mentions: The physiological distinctions observed in response to Cd stress (Figs. 1 and 2, Additional file 1: Figure S1, Additional file 2: Figure S2 and Additional file 3: Figure S3) between the two unique barley genotypes led to further exploitation of their differences at the transcriptome level. Overall, we found that compared to the control plants, the gene expression profiles the leaves of two genotypes were significantly altered after 15 days in 5 μM Cd. Cadmium stress induced differential expression of 812 genes. Among these genes, 382 (131) and 303 (106) were up-regulated (down-regulated) in W6nk2 and Zhenong8, respectively, with a fold change of at least 2.0 (P ≤ 0.05). These genes represented 2.3 and 1.8 % of the total genes in the two genotypes (data not shown). There were distinct differences in gene expression between the two genotypes (Additional file 4: Figure S4, Additional file 5: Figure S5 and Fig. 5; Table 1, Additional file 6: Table S1, Additional file 7: Table S2, Additional file 8: Table S3 and Additional file 9: Table S4), with 338 genes being up-regulated in W6nk2 (W-up) but unchanged in Zhenong8 (Z-NC), while another 13 genes were up-regulated in W6nk2 (W-up), but down-regulated in Zhenong8 (Z-down) (Additional file 4: Figure S4A).Table 1

Bottom Line: Quantitative RT-PCR confirmed our microarray data.Novel transporter genes such as HvZIP3 and HvZIP8 were identified as being associated with low-grain-Cd-accumulation.In addition to advancing academic knowledge, our findings may also result in potential economic benefits for molecular breeding of low Cd accumulating barley and other crops.

View Article: PubMed Central - PubMed

Affiliation: Department of Agronomy, College of Agriculture and Biotechnology, Zijingang Campus, Zhejiang University, Hangzhou, 310058, PR China. sunhongyan-8@163.com.

ABSTRACT

Background: Understanding the mechanism of low Cd accumulation in crops is crucial for sustainable safe food production in Cd-contaminated soils.

Results: Confocal microscopy, atomic absorption spectrometry, gas exchange and chlorophyll fluorescence analyses revealed a distinct difference in Cd accumulation and tolerance between the two contrasting barley genotypes: W6nk2 (a low-grain-Cd-accumulating and Cd-sensitive genotype) and Zhenong8 (a high-grain-Cd-accumulating and tolerant genotype). A DNA microarray analysis detected large-scale changes of gene expression in response to Cd stress with a substantial difference between the two genotypes. Cd stress led to higher expression of genes involved in transport, carbohydrate metabolism and signal transduction in the low-grain-Cd-accumulating genotype. Novel transporter genes such as zinc transporter genes were identified as being associated with low Cd accumulation. Quantitative RT-PCR confirmed our microarray data. Furthermore, suppression of the zinc transporter genes HvZIP3 and HvZIP8 by RNAi silencing showed increased Cd accumulation and reduced Zn and Mn concentrations in barley grains. Thus, HvZIP3 and HvZIP8 could be candidate genes related to low-grain-Cd-accumulation.

Conclusion: Novel transporter genes such as HvZIP3 and HvZIP8 were identified as being associated with low-grain-Cd-accumulation. In addition to advancing academic knowledge, our findings may also result in potential economic benefits for molecular breeding of low Cd accumulating barley and other crops.

No MeSH data available.


Related in: MedlinePlus