Silicon efflux transporters isolated from two pumpkin cultivars contrasting in Si uptake

The accumulation of silicon (Si) differs greatly with plant species and cultivars due to different ability of the roots to take up Si. In Si accumulating plants such as rice, barley and maize, Si uptake is mediated by the influx (Lsi1) and efflux (Lsi2) transporters. Here we report isolation and functional analysis of two Si efflux transporters (CmLsi2-1 and CmLsi2-2) from two pumpkin (Cucurbita moschata Duch.) cultivars contrasting in Si uptake. These cultivars are used for rootstocks of bloom and bloomless cucumber, respectively. Different from mutations in the Si influx transporter CmLsi1, there was no difference in the sequence of either CmLsi2 between two cultivars. Both CmLsi2-1 and CmLsi2-2 showed an efflux transport activity for Si and they were expressed in both the roots and shoots. These results confirm our previous finding that mutation in CmLsi1, but not in CmLsi2-1 and CmLsi2-2 are responsible for bloomless phenotype resulting from low Si uptake.Key words: silicon, efflux transporter, pumpkin, cucumber, bloomSilicon (Si) is the second most abundant elements in earth''s crust.¹ Therefore, all plants rooting in soils contain Si in their tissues. However Si accumulation in the shoot differs greatly among plant species, ranging for 0.1 to 10% of dry weight.^1–3 In higher plants, only Poaceae, Equisetaceae and Cyperaceae show a high Si accumulation.^2,3 Si accumulation also differs with cultivars within a species.^4,5 These differences in Si accumulation have been attributed to the ability of the roots to take up Si.^6,7Genotypic difference in Si accumulation has been used to produce bloomless cucumber (Cucumis sativus L.).⁸ Bloom (white and fine powders) on the surface of cucumber fruits is primarily composed of silica (SiO₂).⁹ However, nowadays, cucumber without bloom (bloomless cucumber) is more popular in Japan due to its more attractive and distinctly shiny appearance. Bloomless cucumber is produced by grafting cucumber on some specific pumpkin (Cucurbita moschata Duch.) cultivars. These pumpkin cultivars used for bloomless cucumber rootstocks have lower silicon accumulation compared with the rootstocks used for producing bloom cucumber.⁹Our study showed that the difference in Si accumulation between bloom and bloomless root stocks of pumpkin cultivars results from different Si uptake by the roots.¹⁰ Si uptake has been demonstrated to be mediated by two different types of transporters (Lsi1 and Lsi2) in rice, barley and maize.^11–15 Lsi1 is an influx transporter of Si, belonging to a NIP subfamily of aquaporin family.^10,11,13,14 This transporter is responsible for transport of Si from external solution to the root cells.¹¹ On the other hand, Lsi2 is an efflux transporter of Si, belonging to putative anion transporter.¹² Lsi2 releases Si from the root cells towards the xylem. Both Lsi1 and Lsi2 are required for Si uptake by the roots.^11,12 To understand the mechanism underlying genotypic difference in Si uptake, we have isolated and functionally characterized an influx Si transporter CmLsi1 from two pumpkin cultivars used for rootstocks of bloomless and bloom cucumber.¹⁰ Sequence analysis showed only two amino acids difference of CmLsi1 between two pumpkin cultivars. However, CmLsi1 from bloom rootstock [CmLsi1(B⁺)] showed transport activity for Si, whereas that from bloomless rootstock [CmLsi1(B⁻)] did not.¹⁰ Furthermore, we found that loss of Si transport activity was caused by one amino acid mutation at the position of 242 (from proline to leucine).¹⁰ This mutation resulted in failure to be localized at the plasma membrane, which is necessary for functioning as an influx transporter. The mutated protein was localized at the ER.¹⁰ Here, we report isolation and expression analysis of Si efflux transporters from two pumpkin cultivars contrasting in Si uptake and accumulation to examine whether Si efflux transporter is also involved in the bloom and bloomless phenotypes.     

HvLsi1 is a silicon influx transporter in barley

Most plants accumulate silicon in their bodies, and this is thought to be important for resistance against biotic and abiotic stresses; however, the molecular mechanisms for Si uptake and accumulation are poorly understood. Here, we describe an Si influx transporter, HvLsi1, in barley. This protein is homologous to rice influx transporter OsLsi1 with 81% identity, and belongs to a Nod26-like major intrinsic protein sub-family of aquaporins. Heterologous expression in both Xenopus laevis oocytes and a rice mutant defective in Si uptake showed that HvLsi1 has transport activity for silicic acid. Expression of HvLsi1 was detected specifically in the basal root, and the expression level was not affected by Si supply. There was a weak correlation between Si uptake and the expression level of HvLsi1 in eight cultivars tested. In the seminal roots, HvLsi1 is localized on the plasma membrane on the distal side of epidermal and cortical cells. HvLsi1 is also located in lateral roots on the plasma membrane of hypodermal cells. These cell-type specificity of localization and expression patterns of HvLsi1 are different from those of OsLsi1. These observations indicate that HvLsi1 is a silicon influx transporter that is involved in radial transport of Si through the epidermal and cortical layers of the basal roots of barley.     

Isolation and functional characterization of CsLsi1, a silicon transporter gene in Cucumis sativus

Cucumber (Cucumis sativus) is a widely grown cucurbitaceous vegetable that exhibits a relatively high capacity for silicon (Si) accumulation, but the molecular mechanism for silicon uptake remains to be clarified. Here we isolated and characterized CsLsi1, a gene encoding a silicon transporter in cucumber (cv. Mch‐4). CsLsi1 shares 55.70 and 90.63% homology with the Lsi1s of a monocot and dicot, rice (Oryza sativa) and pumpkin (Cucurbita moschata), respectively. CsLsi1 was predominantly expressed in the roots, and application of exogenous silicon suppressed its expression. Transient expression in cucumber protoplasts showed that CsLsi1 was localized in the plasma membrane. Heterologous expression in Xenopus laevis oocytes showed that CsLsi1 evidenced influx transport activity for silicon but not urea or glycerol. Expression of cucumber CsLsi1‐mGFP under its own promoter showed that CsLsi1 was localized at the distal side of the endodermis and the cortical cells in the root tips as well as in the root hairs near the root tips. Heterologous expression of CsLsi1 in a rice mutant defective in silicon uptake and the over‐expression of this gene in cucumber further confirmed the role of CsLsi1 in silicon uptake. Our results suggest that CsLsi1 is a silicon influx transporter in cucumber. The cellular localization of CsLsi1 in cucumber roots is different from that in other plants, implying the possible effect of transporter localization on silicon uptake capability.     

Tomato roots have a functional silicon influx transporter but not a functional silicon efflux transporter

Silicon (Si) accumulation in shoots differs greatly with plant species, but the molecular mechanisms for this interspecific difference are unknown. Here, we isolated homologous genes of rice Si influx (SlLsi1) and efflux (SlLsi2) transporter genes in tomato (Solanum lycopersicum L.) and functionally characterized these genes. SlLsi1 showed transport activity for Si when expressed in both rice lsi1 mutant and Xenopus laevis oocytes. SlLsi1 was constitutively expressed in the roots. Immunostaining showed that SlLsi1 was localized at the plasma membrane of both root tip and basal region without polarity. Furthermore, overexpression of SlLsi1 in tomato increased Si concentration in the roots and root cell sap but did not alter the Si concentration in the shoots. By contrast, two Lsi2-like proteins did not show efflux transport activity for Si in Xenopus oocytes. However, when functional CsLsi2 from cucumber was expressed in tomato, the Si uptake was significantly increased, resulting in higher Si accumulation in the leaves and enhanced tolerance of the leaves to water deficit and high temperature. Our results suggest that the low Si accumulation in tomato is attributed to the lack of functional Si efflux transporter Lsi2 required for active Si uptake although SlLsi1 is functional.     

High-resolution secondary ion mass spectrometry reveals the contrasting subcellular distribution of arsenic and silicon in rice roots

Rice (Oryza sativa) takes up arsenite mainly through the silicic acid transport pathway. Understanding the uptake and sequestration of arsenic (As) into the rice plant is important for developing strategies to reduce As concentration in rice grain. In this study, the cellular and subcellular distributions of As and silicon (Si) in rice roots were investigated using high-pressure freezing, high-resolution secondary ion mass spectrometry, and transmission electron microscopy. Rice plants, both the lsi2 mutant lacking the Si/arsenite efflux transporter Lsi2 and its wild-type cultivar, with or without an iron plaque, were treated with arsenate or arsenite. The formation of iron plaque on the root surface resulted in strong accumulation of As and phosphorous on the epidermis. The lsi2 mutant showed stronger As accumulation in the endodermal vacuoles, where the Lsi2 transporter is located in the plasma membranes, than the wild-type line. As also accumulated in the vacuoles of some xylem parenchyma cells and in some pericycle cells, particularly in the wild-type mature root zone. Vacuolar accumulation of As is associated with sulfur, suggesting that As may be stored as arsenite-phytochelatin complexes. Si was localized in the cell walls of the endodermal cells with little apparent effect of the Lsi2 mutation on its distribution. This study reveals the vacuolar sequestration of As in rice roots and contrasting patterns of As and Si subcellular localization, despite both being transported across the plasma membranes by the same transporters.     

Identification and Characterization of Maize and Barley Lsi2-Like Silicon Efflux Transporters Reveals a Distinct Silicon Uptake System from That in Rice

Silicon (Si) uptake has been extensively examined in rice (Oryza sativa), but it is poorly understood in other gramineous crops. We identified Low Silicon Rice 2 (Lsi2)-like Si efflux transporters from two important gramineous crops: maize (Zea mays) and barley (Hordeum vulgare). Both maize and barley Lsi2 expressed in Xenopus laevis oocytes showed Si efflux transport activity. Furthermore, barley Lsi2 was able to recover Si uptake in a rice mutant defective in Si efflux. Maize and barley Lsi2 were only expressed in the roots. Expression of maize and barley Lsi2 was downregulated in response to exogenously applied Si. Moreover, there was a significant positive correlation between the ability of roots to absorb Si and the expression levels of Lsi2 in eight barley cultivars, suggesting that Lsi2 is a key Si transporter in barley. Immunostaining showed that maize and barley Lsi2 localized only at the endodermis, with no polarity. Protein gel blot analysis indicated that maize and barley Lsi2 localized on the plasma membrane. The unique features of maize and barley Si influx and efflux transporters, including their cell-type specificity and the lack of polarity of their localization in Lsi2, indicate that these crops have a different Si uptake system from that in rice.     

植物对硅的吸收转运机制研究进展

硅(Si)能缓解生物与非生物胁迫对植物的毒害作用,Si的吸收转运是由Si转运蛋白介导的.最近,多个Si转运蛋白(Lsi)基因相继在水稻、大麦和玉米中被克隆出来,并在Si的吸收转运机制方面取得了很大进展.水稻OsLsi在根组织中呈极性分布,OsLsi1定位在根外皮层和内皮层凯氏带细胞外侧质膜,负责将外部溶液中的单硅酸转运到皮层细胞内.OsLsi2定位在凯氏带细胞内侧质膜,在外皮层中负责将Si输出到通气组织质外体中,在内皮层与OsLsi1协同作用将Si转运到中柱中.导管中的Si通过蒸腾流转运到地上部,再由定位在叶鞘和叶片木质部薄壁细胞靠近导管一侧的OsLsi6负责木质部Si的卸载和分配.在大麦和玉米中,ZmLsi1/HvLsi1定位在根表皮和皮层细胞外侧质膜负责Si的吸收,然后Si通过共质体途径被转运到内皮层凯氏带细胞中,再由ZmLsi2/HvLsi2输出转运到中柱中.ZmLsi6在细胞中的定位和活性与OsLsi6相似,推测其可能具有类似的功能,但大麦Lsi6至今未见报道.所以,Si转运机制仍需要进一步研究.     

Identification of maize silicon influx transporters

Maize (Zea mays L.) shows a high accumulation of silicon (Si),but transporters involved in the uptake and distribution havenot been identified. In the present study, we isolated two genes(ZmLsi1 and ZmLsi6), which are homologous to rice influx Sitransporter OsLsi1. Heterologous expression in Xenopus laevisoocytes showed that both ZmLsi1 and ZmLsi6 are permeable tosilicic acid. ZmLsi1 was mainly expressed in the roots. By contrast,ZmLsi6 was expressed more in the leaf sheaths and blades. Differentfrom OsLsi1, the expression level of both ZmLsi1 and ZmLsi6was unaffected by Si supply. Immunostaining showed that ZmLsi1was localized on the plasma membrane of the distal side of rootepidermal and hypodermal cells in the seminal and crown roots,and also in cortex cells in lateral roots. In the shoots, ZmLsi6was found in the xylem parenchyma cells that are adjacent tothe vessels in both leaf sheaths and leaf blades. ZmLsi6 inthe leaf sheaths and blades also exhibited polar localizationon the side facing towards the vessel. Taken together, it canbe concluded that ZmLsi1 is an influx transporter of Si, whichis responsible for the transport of Si from the external solutionto the root cells and that ZmLsi6 mainly functions as a Si transporterfor xylem unloading.     

Uptake system of silicon in different plant species

The accumulation of silicon (Si) in the shoots varies considerably among plant species, but the mechanism responsible for this variation is poorly understood. The uptake system of Si was investigated in terms of the radial transport from the external solution to the root cortical cells and the release of Si from the cortical cells to the xylem in rice, cucumber, and tomato, which differ greatly in shoot Si concentration. Symplasmic solutions of the root tips were extracted by centrifugation. The concentrations of Si in the root-cell symplast in all species were higher than that in the external solution, although the concentration in rice was 3- and 5-fold higher than that in cucumber and tomato, respectively. A kinetic study showed that the radial transport of Si was mediated by a transporter with a K(m) value of 0.15 mM in all species, but with different V(max) values in the order of rice>cucumber>tomato. In the presence of the metabolic inhibitor 2,4-dinitrophenol, and at low temperature, the Si concentration in the root-cell symplast decreased to a level similar to that of the apoplasmic solution. These results suggest that both transporter-mediated transport and passive diffusion of Si are involved in the radial transport of Si and that the transporter-mediated transport is an energy-dependent process. The Si concentration of xylem sap in rice was 20- and 100-fold higher than that in cucumber and tomato, respectively. In contrast to rice, the Si concentration in the xylem sap was lower than that in the external solution in cucumber and tomato. A kinetic study showed that xylem loading of Si was also mediated by a kind of transporter in rice, but by passive diffusion in cucumber and tomato. These results indicate that a higher density of transporter for radial transport and the presence of a transporter for xylem loading are responsible for the high Si accumulation in rice.     

Improving the Mineral Nutrition in Grafted Watermelon Plants: Nitrogen Metabolism

Watermelon (Citrullus lanatus [Trumb.] Mansfeld cv. Early Star), was used as scion grafted onto three cultivars of pumpkin (Cucurbita pepo L. cvs. Brava, Shintoza and Kamel) used as rootstocks and ungrafted Early Star plants were used as control. The rootstocks showed a high capacity for N uptake and transport to the scion where N reduction and assimilation improved growth of the scion in grafted plants with respect to the control.     

A transporter regulating silicon distribution in rice shoots

Rice (Oryza sativa) accumulates very high concentrations of silicon (Si) in the shoots, and the deposition of Si as amorphous silica helps plants to overcome biotic and abiotic stresses. Here, we describe a transporter, Lsi6, which is involved in the distribution of Si in the shoots. Lsi6 belongs to the nodulin-26 intrinsic protein III subgroup of aquaporins and is permeable to silicic acid. Lsi6 is expressed in the leaf sheath and leaf blades as well as in the root tips. Cellular localization studies revealed that Lsi6 is found in the xylem parenchyma cells of the leaf sheath and leaf blades. Moreover, Lsi6 showed polar localization at the side facing toward the vessel. Knockdown of Lsi6 did not affect the uptake of Si by the roots but resulted in disordered deposition of silica in the shoots and increased excretion of Si in the guttation fluid. These results indicate that Lsi6 is a transporter responsible for the transport of Si out of the xylem and subsequently affects the distribution of Si in the leaf.     

Subcellular Localization of Zinc and Calcium in Bean (Phaseolus vulgaris L.) Tissues

Two bean (Phaseolus vulgaris L.) cultivars differing in growth responses to zinc were examined for differences in uptake and subcellular localization of ⁶⁵Zn during a 15-day growth period. The zinc-sensitive cultivar Sanilac showed initially a much higher rate of absorption, which declined after 24 hours. The zinc-tolerant cultivar Saginaw showed a slow but steady rate of absorption for 10 days. In roots as well as in stem callus tissues of both cultivars, three-fourths of the absorbed ⁶⁵Zn was localized in the “cytoplasmic” supernatant fractions (containing ribosomes and vacuolar sap). Very little (less than 7%) ⁶⁵Zn was localized in the cell wall fraction. There was a much greater proportion of the absorbed ⁶⁵Zn localized in root mitochondria and nuclei of the zinc-sensitive Sanilac than in the zinc-tolerant Saginaw. Stem callus tissues, however, did not show such cultivar differences in zinc accumulation at the sub-cellular level.     

Spatial distribution and temporal variation of the rice silicon transporter Lsi1

Rice (Oryza sativa) is a typical silicon (Si) accumulator and requires a large amount of Si for high-yield production. Recently, a gene (Low silicon rice1 [Lsi1]) encoding a Si transporter was identified in rice roots. Here, we characterized Lsi1 in terms of spatial distribution and temporal variation using both physiological and molecular approaches. Results from a multicompartment transport box experiment showed that the major site for Si uptake was located at the basal zone (>10 mm from the root tip) of the roots rather than at the root tips (<10 mm from the root tip). Consistent with the Si uptake pattern, Lsi1 expression and distribution of the Lsi1 protein were found only in the basal zone of roots. In the basal zones of the seminal, crown, and lateral roots, the Lsi1 protein showed a polar localization at the distal side of both the exodermis and endodermis, where the Casparian bands are formed. This indicates that Lsi1 is required for the transport of Si through the cells of the exodermis and endodermis. Expression of Lsi1 displayed a distinct diurnal pattern. Furthermore, expression was transiently enhanced around the heading stage, which coincides with a high Si requirement during this growth stage. Expression was down-regulated by dehydration stress and abscisic acid, suggesting that expression of Lsi1 may be regulated by abscisic acid.     

Analysis of subcellular localization of auxin carriers PIN,AUX/LAX and PGP in Sorghum bicolor

Auxin transport at least correlates to the three gene families: efflux carriers PIN-formed (PIN), p-glycoprotein (PGP), and influx carrier auxin resistant 1/like aux1(AUX/LAX) in Arabidopsis thaliana. In monocotyledon Sorghum bicolor, the biological function of these genes retains unclear. Our previous study reported that the member analysis, organ-specific expression and expression profiles of the auxin transporter PIN, PGP and AUX/LAX gene families in Sorghum bicolor under IAA, brassinosteroid, polar auxin transport inhibitors and abiotic stresses. Here we further supply the prediction of subcellular localization of SbPIN, SbLAX and SbPGP proteins and discuss the potential relationship between the subcellular localization and stress response. The predicted results showed that the most of SbPIN, SbLAX and SbPGP proteins are localized to the plasma membrane, except few localized to vacuolar membrane and endoplasmic reticulum. This data set provides novel information for investigation of auxin transporters in Sorghum bicolor.     

Influence of rootstocks on growth,yield, fruit quality and leaf mineral element contents of pear cv. ‘Santa Maria’ in semi-arid conditions

Background

Rootstocks play an essential role to determining orchard performance of fruit trees. Pyrus communis and Cydonia oblonga are widely used rootstocks for European pear cultivars. The lack of rootstocks adapted to different soil conditions and different grafted cultivars is widely acknowledged in pear culture. Cydonia rootstocks (clonal) and Pyrus rootstocks (seedling or clonal) have their advantages and disadvantages. In each case, site-specific environmental characteristics, specific cultivar response and production objectives must be considered before choosing the best rootstock. In this study, the influence of three Quince (BA 29, Quince A = MA, Quince C = MC) and a local European pear seedling rootstocks on the scion yield, some fruit quality characteristics and leaf macro (N, P, K, Ca and Mg) and micro element (Fe, Zn, Cu, Mn and B) content of ‘Santa Maria’ pear (Pyrus communis L.) were investigated.

Results

Trees on seedling rootstock had the highest annual yield, highest cumulative yield (kg tree⁻¹), largest trunk cross-sectional area (TCSA), lowest yield efficiency and lowest cumulative yield (ton ha⁻¹) in the 10^th year after planting. The rootstocks had no significant effect on average fruit weight and fruit volume. Significantly higher fruit firmness was obtained on BA 29 and Quince A. The effect of rootstocks on the mineral element accumulation (N, K, Ca, Mg, Fe, Zn, Cu, Mn and B) was significant. Leaf analysis showed that rootstocks used had different mineral uptake efficiencies throughout the early season.

Conclusion

The results showed that the rootstocks strongly affected fruit yield, fruit quality and leaf mineral element uptake of ‘Santa Maria’ pear cultivar. Pear seedling and BA 29 rootstock found to be more prominent in terms of several characteristics for ‘Santa Maria’ pear cultivar that is grown in highly calcareous soil in semi-arid climate conditions. We determined the highest N, P (although insignificant), K, Ca, Mg, Fe and Cu mineral element concentrations on the pear seedling and BA 29 rootstocks. According to the results, we recommend the seedling rootstock for normal density plantings (400 trees ha⁻¹) and BA 29 rootstock for high-density plantings (800 trees ha⁻¹) for ‘Santa Maria’ pear cultivar in semi-arid conditions.     

Cl‐ homeostasis in includer and excluder citrus rootstocks: transport mechanisms and identification of candidate genes

To reveal specific Cl^‐ transport activities in the symplastic pathway, uptake, long‐distance transport and distribution of Cl^‐ have been investigated in the citrus rootstocks Carrizo citrange (CC, Cl^‐ includer) and Cleopatra mandarin (CM, Cl^‐ excluder). Using an external concentration of 4.5 mm Cl^‐, both species actively transported Cl^‐ to levels that exceeded the critical requirement concentration by one and two orders of magnitude in the excluder and the includer rootstocks, respectively. Both CC and CM modulated Cl^‐ influx according to the availability of the nutrient as uptake capacity was induced by Cl^‐ starvation, but inhibited after Cl^‐ resupply. Net Cl^‐ uptake was higher in the includer CC, an observation that correlated with a lower root‐to‐shoot transport capacity in the excluder CM. The patterns of tissue Cl^‐ accumulation indicated that chloride exclusion in the salt‐tolerant rootstock CM was caused by a reduced net Cl^‐ loading into the root xylem. Genes CcCCC1, CcSLAH1 and CcICln1 putatively involved in the regulation of chloride transport were isolated and their expression analysed in response to both changes in the nutritional status of Cl^‐ and salt stress. The previously uncharacterized ICln gene exhibited a strong repression to Cl^‐ application in the excluder rootstock, suggesting a role in regulating Cl^‐ homeostasis in plants.     

Variation in the content of bioflavonoids of orange as affected by scion,rootstock, and fruit part

Fruits of citrus cultivars contain bioflavonoids and some other important secondary metabolites in pharmaceutical and nutritional industries. The present experiment was designed to investigate the correlation between the content of flavonoid components like naringin, hesperidin, and neohesperidin of the scions fruits and the same parameters in rootstocks fruits. Six-year-old trees including four citrus cultivars of ‘Moro’, ‘Mars’, ‘Salustiana’, and ‘Italian’ which were previously grafted on the four different rootstocks including ‘Yuzu’, ‘Shelmahalleh’, ‘Citromelo’, and ‘Sour orange’ were selected as experimental trees. The content of the mentioned flavonoids was investigated in the peel and pulp of the fruits of both scions and rootstocks. The results showed that the measured parameters were significantly influenced by scions, rootstocks, and tissues. Based on the obtained results, it can be suggested that the accumulation of chemicals in citrus fruit depends on genetic and inherent abilities of the scion, more than what was previously believed, while the rootstock can also play an important role in the accumulation of these substances.     

Silicon alleviates cadmium toxicity in peanut plants in relation to cadmium distribution and stimulation of antioxidative enzymes

Silicon (Si) is generally considered a beneficial element for the growth of higher plants, especially for those grown under stressed environments. Recently, the mitigating role of Si in cadmium (Cd) stress has received some attention. However, its mechanisms involved remain poorly understood. We studied the effects of Si on tissue and subcellular distribution of Cd, as well as the activities of major antioxidant enzymes (SOD, POD and CAT) with two contrasting peanut (Arachis hypogaea L.) cultivars (Luhua 11 and Luzi 101) differing in their Cd tolerance. The results showed that Cd exposure alone depressed plant growth and caused oxidative stress for both cultivars, and this toxicity was more obvious in Cd-sensitive cultivar (Luhua 11) than in Cd-tolerant cultivar (Luzi 101). Si supply significantly alleviated the toxicity of Cd in peanut seedlings; this was correlated with a reduction of shoot Cd accumulation, an alteration of Cd subcellular distribution in leaves, and a stimulation of antioxidative enzymes. The mechanisms of Si amelioration of Cd stress were cultivar and tissue dependent. For Luhua 11, Si-mediated inhibition of Cd transport from roots to shoots, reduction of Cd content in cell organelle fractions of leaves, and enhancement of the SOD, POD and CAT activities in roots, might responsible for the role of Si in alleviating Cd toxicity. For Luzi 101, Si alleviation of Cd toxicity is mainly attributed to the decrease in Cd concentration in shoot and stimulation of antioxidants systems.     

Improving magnesium uptake,photosynthesis and antioxidant enzyme activities of watermelon by grafting onto pumpkin rootstock under low magnesium

Background and aims

Magnesium (Mg) is an essential macronutrient that plays an important role in numerous physiological and biochemical processes of plant. However, Mg deficiency commonly occurs worldwide. Watermelon is an important crop that often suffers from Mg deficiency. This study aims to test whether watermelon performance can be improved by grafting onto rootstocks under low Mg and to clarify the underlying physiological mechanism.

Methods

Self-grafted, bottle gourd (Jingxinzhen No.1) and pumpkin (Jingxinzhen No.4) rootstock-grafted plants were treated with three Mg concentrations: 2.0 mM (normal condition), 0.4 mM (moderate stress), and 0.04 mM (severe stress) for 16 days under hydroponic conditions. Ungrafted watermelon and pumpkin were treated with 2.0 mM and 0.04 mM for 12 days.

Results

The growth of the plants was not affected by 0.4 mM Mg; however, plant growth decreased under 0.04 mM Mg in all graft combinations compared with control (2.0 mM Mg). Pumpkin rootstock grafting significantly increased watermelon growth under low Mg stress (0.04 mM Mg), compared with self-grafted and bottle gourd-grafted plants. The Mg²⁺ uptake of watermelon plants was increased by grafting onto pumpkin rootstocks, however, root-to-shoot transport capacity of Mg²⁺ was similar compared with self-grafted plants under 0.04 mM Mg. Gene expression analysis showed that magnesium transporter genes MGT1, MGT3, MGT4, and MGT5 may play an important role in higher Mg²⁺ uptake of pumpkin root. The photosynthetic parameters and activities of superoxide dismutase, peroxidase and catalase were significantly higher, but malonaldehyde (MDA) content were lower in the pumpkin rootstock grafted plants compared with other graft combinations under 0.04 mM Mg.

Conclusion

Our results provide strong evidence that pumpkin rootstock ‘Jinxinzhen No. 4’ grafting can improve watermelon performance under low Mg stress. The enhanced plant performance is attributed to higher root Mg²⁺ uptake and the improvement of photosynthesis and antioxidant enzyme activities.