A Pleiotropic Drug Resistance Family Protein Gene Is Required for Rice Growth, Seed Development and Zinc Homeostasis

doi:10.1016/j.rsci.2023.01.005

Abstract

Abstract:

Zinc (Zn) is an essential mineral element for plant growth and development. Zn deficiency in crops frequently occurs in many types of soils. It is therefore crucial to identify genetic resources linking Zn acquisition traits and development of crops with improved Zn-use efficiency for sustainable crop production. In this study, we functionally identified a rice uncharacterized ABCG (ATP-binding cassette G-subfamily) gene encoding a PDR20 (pleiotropic drug resistance 20) metal transporter for mediation of rice growth, seed development and Zn accumulation. OsPDR20 was localized to the plasma membrane, but it was not transcriptionally induced under Zn deficiency, rather was sufficiently up-regulated under high level of Zn stress. Yeast (Saccharomyces cerevisiae) transformed with OsPDR20 displayed a relatively lower Zn accumulation with attenuated cellular growth, suggesting that OsPDR20 had an activity for Zn transport. Knocking-down OsPDR20 by RNA interference (RNAi) compromised rice growth with shorter plant height and decreased biomass in rice plantlets grown under hydroponic media. Zn concentration in the roots of OsPDR20 knocked-down rice lines declined under Zn deficiency, while they remained unchanged compared with the wild type under normal Zn supply. A rice lifelong field trial demonstrated that OsPDR20 mutation impaired the capacity of seed development, with shortened panicle and seed length, compromised spikelet fertility, and reduced grain number per plant or grain weight per unit area. Interestingly, OsPDR20 mutation elevated the accumulation of Zn in husk and brown rice over the wild type. Overall, this study pointed out that OsPDR20 is fundamentally required for rice growth and seed development through Zn transport and homeostasis.

Key words: OsPDR20, zinc transport, rice, seed development, ABCG53, pleiotropic drug resistance

Li Chao, Li He, Zhang Xianduo, Yang Zhimin. A Pleiotropic Drug Resistance Family Protein Gene Is Required for Rice Growth, Seed Development and Zinc Homeostasis[J]. Rice Science, 2023, 30(2): 127-137.

Figures/Tables 7

Fig. 1. Expression patterns of OsPDR20 in wild type rice under Zn deficient or overloaded stress by qRT-PCR. A and B, Changes of OsPDR20 transcripts in root (A) and shoot (B) tissues were monitored during the course of Zn deprivation. C, Transcripts of OsPDR20 were determined at 0.4 (control), 500, 1 000 and 5 000 μmol/L Zn concentrations for 6 h. Two-week-old rice was exposed to normal Zn supply (+Zn) and Zn deficiency (-Zn) for 14 d. Data are Mean ± SD (n = 3). The asterisks indicate the significant difference between the control and treatment (P < 0.05).

Fig. 2. Assessment of subcellular localization of OsPDR20 by expression in rice mesophyll protoplasts and tobacco epidermal cells. A and B, Fluorescence of pAN580-GFP as control vector (A) and OsPDR20-pAN580-GFP fusion protein (B) that were transiently transformed into rice mesophyll protoplasts. BF, Bright field; GFP, Green fluorescent protein. C, Fluorescence of OsPDR20-pAN580-GFP in tobacco epidermal cells. PIP2A-RFP was used as a biomarker for plasma membrane location.

Fig. 3. Growth phenotypes of wild type (WT) rice and its RNAi lines (i-1, i-2 and i-6) under Zn-normal supply (+Zn) and Zn-deficiency (-Zn) conditions. A-D, Phenotypes of WT and RNAi lines under +Zn and -Zn conditions. Scale bars are 10 cm. E and F, Shoot lengths (E) and root lengths (F) of WT and RNAi lines. G and H, Dry weights (DW) of shoot (G) and root (H) tissues in WT and RNAi lines. Two-week-old rice was hydroponically grown for 28 d. Data are Mean ± SD (n = 3). The asterisks indicate the significant difference between WT and RNAi lines (P < 0.05).

Fig. 4. Zn concentrations in root and shoot tissues of wild type (WT) and RNAi lines (i-1, i-2 and i-6) with Zn translocation from roots to shoots under Zn-normal supply (+Zn) and Zn-deficiency (-Zn) conditions. A and B, Zn concentrations in roots (A) and shoots (B). C, Translocation of Zn from roots to shoots. Two-week-old rice plants were hydroponically grown for 28 d. Data are Mean ± SD (n = 3). The asterisks indicate the significant difference between WT and RNAi lines (P < 0.05).

Fig. 5. Morphological differences between wild type (WT) and RNAi lines (i-1, i-2 and i-6) at the maturity stage. A, Phenotypes of WT and RNAi plants. Scale bar is 10 cm. B, Plant heights of WT and RNAi lines. C, Main stem structures of WT and RNAi lines. The distance between the arrows represents the length of internode. Scale bar is 10 cm. D, Internode lengths of WT and RNAi lines. E, Tiller number per plant of WT and RNAi lines. F, Effective panicle number per plant of WT and RNAi lines. The rice plants were grown to maturity under natural conditions. Data in B, D, E and F are Mean ± SD (n = 3). The asterisks indicate the significant difference between WT and RNAi lines (P < 0.05).

Fig. 6. Agronomic traits relevant to seed development of wild type (WT) and RNAi lines (i-1, i-2 and i-6). A and B, Phenotypes of panicle length. Scale bar is 3 cm. C and D, Phenotypes of grain width. Scale bar is 1 cm. E and F, Phenotypes of grain length. Scale bar is 1 cm. G, Seed weight per plant. H, Grain yield per square meter. Data are Mean ± SD (n = 3). The asterisks indicate the significant difference between WT and RNAi lines (P < 0.05).

Fig. 7. Zn concentrations in different tissues or organs of wild type (WT) and RNAi lines (i-1, i-2 and i-6) at the maturity stage. A, Assessment of Zn concentrations in rice straw (upper and lower leaves and stem). B, Calculation of Zn translocation from leaves to grains (husk and brown rice). Rice was grown to the seed maturity stage under natural conditions for two consecutive years. Data are Mean ± SD (n = 3). The asterisks indicate the significant difference between WT and RNAi lines (P < 0.05).

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