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Background: Castor is an important industrial raw material. Drought-induced oxidative stress leads to slow growth and decreased yields in castor. However, the mechanisms of drought-induced oxidative stress in castor remain unclear. Therefore, in this study, physiological, biochemical, and RNA-seq analyses were conducted on the roots of castor plants under PEG-6000 stress for 3 d and 7 d followed by 4 d of hydration., Results: The photosynthetic rate of castor leaves was inhibited under PEG-6000 stress for 3 and 7 d. Biochemical analysis of castor roots stressed for 3 d and 7 d, and rehydrated for 4 d revealed that the activities of APX and CAT were highest after only 3 d of stress, whereas the activities of POD, GR, and SOD peaked after 7 d of stress. RNA-seq analysis revealed 2926, 1507, and 111 differentially expressed genes (DEGs) in the roots of castor plants under PEG-6000 stress for 3 d and 7 d and after 4 d of rehydration, respectively. GO analysis of the DEGs indicated significant enrichment in antioxidant activity. Furthermore, KEGG enrichment analysis of the DEGs revealed significantly enriched metabolic pathways, including glutathione metabolism, fatty acid metabolism, and plant hormone signal transduction. WGCNA identified the core genes PP2C39 and GA2ox4 in the navajowhite1 module, which was upregulated under PEG-6000 stress. On the basis of these results, we propose a model for the response to drought-induced oxidative stress in castor., Conclusions: This study provides valuable antioxidant gene resources, deepening our understanding of antioxidant regulation and paving the way for further molecular breeding of castor plants., (© 2024. The Author(s).)

Castor (Ricinus communis L.) is an importance crop cultivated for its oil and economic value. Seed size is a crucial factor that determines crop yield. Gaining insight into the molecular regulatory processes of seed development is essential for the genetic enhancement and molecular breeding of castor. Here, we successfully fine-mapped a major QTL related to seed size, qSS3, to a 180 kb interval on chromosome 03 using F2 populations (DL01×WH11). A 17.6-kb structural variation (SV) was detected through genomic comparison between DL01 and WH11. Analysis of haplotypes showed that the existence of the complete 17.6 kb structural variant may lead to the small seed characteristic in castor. In addition, we found that qSS3 contains the microRNA396b (miR396b) sequence, which is situated within the 17.6 kb SV. The results of our experiment offer additional evidence that miR396-Growth Regulating Factor 4 (GRF4) controls seed size by impacting the growth and multiplication of seed coat and endosperm cells. Furthermore, we found that RcGRF4 activates the expression of YUCCA6 (YUC6), facilitating the production of IAA in seeds and thereby impacting the growth of castor seeds. Our research has discovered a crucial functional module that controls seed size, offering a fresh understanding of the mechanism underlying seed size regulation in castor., Competing Interests: Conflict of interest statement. The authors declare that there is no conflict of interest., (© The Author(s) 2024. Published by Oxford University Press on behalf of American Society of Plant Biologists. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Genomic imprinting refers to parent-of-origin-dependent gene expression and primarily occurs in the endosperm of flowering plants, but its functions and epigenetic mechanisms remain to be elucidated in eudicots. Castor bean, a eudicot with large and persistent endosperm, provides an excellent system for studying the imprinting. Here, we identified 131 imprinted genes in developing endosperms and endosperm at seed germination phase of castor bean, involving into the endosperm development, accumulation of storage compounds and specially seed germination. Our results showed that the transcriptional repression of maternal allele of DNA METHYLTRANSFERASE 1 (MET1) may be required for maternal genome demethylation in the endosperm. DNA methylation analysis showed that only a small fraction of imprinted genes was associated with allele-specific DNA methylation, and most of them were closely associated with constitutively unmethylated regions (UMRs), suggesting a limited role for DNA methylation in controlling genomic imprinting. Instead, histone modifications can be asymmetrically deposited in maternal and paternal genomes in a DNA methylation-independent manner to control expression of most imprinted genes. These results expanded our understanding of the occurrence and biological functions of imprinted genes and showed the evolutionary flexibility of the imprinting machinery and mechanisms in plants., (© 2023 The Authors. New Phytologist © 2023 New Phytologist Foundation.)

Castor bean (Ricinus communis L.) can withstand long periods of water deficit and high temperatures, and therefore has been recognized as a drought-resistant plant species, allowing the study of gene networks involved in drought response and tolerance. The identification of genes networks related to drought response in this plant may yield important information in the characterization of molecular mechanisms correlating changes in the gene expression with the physiological adaptation processes. In this context, gene families related to abscisic acid (ABA) signaling play a crucial role in developmental and environmental adaptation processes of plants to drought stress. However, the families that function as the core components of ABA signaling, as well as genes networks related to drought response, are not well understood in castor bean. In this study 7 RcPYL, 63 RcPP2C, and 6 RcSnRK2 genes were identified in castor bean genome, which was further supported by chromosomal distribution, gene structure, evolutionary relationships, and conserved motif analyses. The castor bean general expression profile was investigated by RNAseq in root and leaf tissues in response to drought stress. These analyses allowed the identification of genes differentially expressed, including genes from the ABA signaling core, genes related to photosynthesis, cell wall, energy transduction, antioxidant response, and transcription factors. These analyses provide new insights into the core components of ABA signaling in castor bean, allow the identification of several molecular responses associated with the high physiological adaptation of castor bean to drought stress, and contribute to the identification of candidate genes for genetic improvement., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Castor (Ricinus communis L.) is a dicotyledonous oilseed crop that can have either spineless or spiny capsules. Spines are protuberant structures that differ from thorns or prickles. The developmental regulatory mechanisms governing spine formation in castor or other plants have remained largely unknown. Herein, using map-based cloning in 2 independent F2 populations, F2-LYY5/DL01 and F2-LYY9/DL01, we identified the RcMYB106 (myb domain protein 106) transcription factor as a key regulator of capsule spine development in castor. Haplotype analyses demonstrated that either a 4,353-bp deletion in the promoter or a single nucleotide polymorphism leading to a premature stop codon in the RcMYB106 gene could cause the spineless capsule phenotype in castor. Results of our experiments indicated that RcMYB106 might target the downstream gene RcWIN1 (WAX INDUCER1), which encodes an ethylene response factor known to be involved in trichome formation in Arabidopsis (Arabidopsis thaliana) to control capsule spine development in castor. This hypothesis, however, remains to be further tested. Nevertheless, our study reveals a potential molecular regulatory mechanism underlying the spine capsule trait in a nonmodel plant species., Competing Interests: Conflict of interest statement. The authors declare that there is no conflict of interest., (© American Society of Plant Biologists 2023. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The rise in oil prices, global warming, and the depletion of nonrenewable resources have led researchers to study sustainable alternatives to increasing energy demand. The autocatalysis from castor oil and castor lipases to produce biodiesel can be an excellent alternative to reduce the production costs and avoid the drawbacks of chemical transesterification. This study aimed to evaluate the catalytic activity of castor bean lipase extract (CBLE) on three vegetable oils hydrolysis, to obtain and enhance biodiesel yield by an autocatalysis from castor oil and CBLE. Furthermore, the enzymatic biodiesel physicochemical quality was analyzed. The enzymatic activity for olive oil was 76.12 U, 90.06 U for commercial castor oil, and 75.60 U in raw castor oil. The hydrolysis percentages were high at 25 °C, pH 4.5, for 4 h with 97.18% for olive oil, 98.86%, and 96.19% for commercial and raw castor oil, respectively. The CBLE catalyzed the transesterification reaction on castor oil to obtain 82.91% biodiesel yield under the selected conditions of 20% lipase loading, 1:6 oil/methanol molar ratio, and 10% buffer pH 4.5, 37 °C for 8 h. The castor biodiesel quality satisfied the ASTM-D6751 (USA) and EN-14214 (European Union) values, except for the density, viscosity, and moisture, as expected for this kind of biodiesel., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Phosphoenolpyruvate carboxylase (PEPC) is a tightly regulated enzyme that plays a crucial anaplerotic role in central plant metabolism. Bacterial-type PEPC (BTPC) of developing castor oil seeds (COS) is highly expressed as a catalytic and regulatory subunit of a novel Class-2 PEPC heteromeric complex. Ricinus communis Ca2+-dependent protein kinase-1 (RcCDPK1) catalyzes in vivo inhibitory phosphorylation of COS BTPC at Ser451. Autokinase activity of recombinant RcCDPK1 was detected and 42 autophosphorylated Ser, Thr or Tyr residues were mapped via liquid chromatography-tandem mass spectrometry. Prior autophosphorylation markedly attenuated the ability of RcCDPK1 to transphosphorylate its BTPC substrate at Ser451. However, fully dephosphorylated RcCDPK1 rapidly autophosphorylated during the initial stages of a BTPC transphosphorylation assay. This suggests that Ca2+-dependent binding of dephospho-RcCDPK1 to BTPC may trigger a structural change that leads to rapid autophosphorylation and subsequent substrate transphosphorylation. Tyr30 was identified as an autophosphorylation site via LC-MS/MS and immunoblotting with a phosphosite-specific antibody. Tyr30 occurs at the junction of RcCDPK1's N-terminal variable (NTVD) and catalytic domains and is widely conserved in plant and protist CDPKs. Interestingly, a reduced rate and extent of BTPC transphosphorylation occurred with a RcCDPK1Y30F mutant. Prior research demonstrated that RcCDPK1's NTVD is essential for its Ca2+-dependent autophosphorylation or BTPC transphosphorylation activities but plays no role in target recognition. We propose that Tyr30 autophosphorylation facilitates a Ca2+-dependent interaction between the NTVD and Ca2+-activation domain that primes RcCDPK1 for transphosphorylating BTPC at Ser451. Our results provide insights into links between the post-translational control of COS anaplerosis, Ca2+-dependent signaling and the biological significance of RcCDPK1 autophosphorylation., (© The Author(s) 2022. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Background: Understanding the processes governing angiosperm seed growth and development is essential both for fundamental plant biology and for agronomic purposes. Master regulators of angiosperm seed development are expressed in a seed-specific manner. However, it is unclear how this seed specificity of transcription is established. In some vertebrates, DNA methylation valleys (DMVs) are highly conserved and strongly associated with key developmental genes, but comparable studies in plants are limited to Arabidopsis and soybean. Castor bean (Ricinus communis) is a valuable model system for the study of seed biology in dicots and source of economically important castor oil. Unlike other dicots such as Arabidopsis and soybean, castor bean seeds have a relatively large and persistent endosperm throughout seed development, representing substantial structural differences in mature seeds. Here, we performed an integrated analysis of RNA-seq, whole-genome bisulfite sequencing, and ChIP-seq for various histone marks in the castor bean., Results: We present a gene expression atlas covering 16 representative tissues and identified 1162 seed-specific genes in castor bean (Ricinus communis), a valuable model for the study of seed biology in dicots. Upon whole-genome DNA methylation analyses, we detected 32,567 DMVs across five tissues, covering ~33% of the castor bean genome. These DMVs are highly hypomethylated during development and conserved across plant species. We found that DMVs have the potential to activate transcription, especially that of tissue-specific genes. Focusing on seed development, we found that many key developmental regulators of seed/endosperm development, including AGL61, AGL62, LEC1, LEC2, ABI3, and WRI1, were located within DMVs. ChIP-seq for five histone modifications in leaves and seeds clearly showed that the vast majority of histone modification peaks were enriched within DMVs, and their remodeling within DMVs has a critical role in the regulation of seed-specific gene expression. Importantly, further experiment analysis revealed that distal DMVs may act as cis-regulatory elements, like enhancers, to activate downstream gene expression., Conclusions: Our results point to the importance of DMVs and special distal DMVs behaving like enhancers, in the regulation of seed-specific genes, via the reprogramming of histone modifications within DMVs. Furthermore, these results provide a comprehensive understanding of the epigenetic regulator roles in seed development in castor bean and other important crops., (© 2022. The Author(s).)

Chelating agents have been considered as an important phytoremediation strategy to enhance heavy metal extraction from contaminated soil. A pot experiment was conducted to explore the effects of low molecular weight organic acids (LMWOAs) on the phytoremediation efficiency of copper (Cu) by castor bean, and soil enzyme activities. Results indicated that the addition of all the three kinds of LMWOAs (citric, tartaric, oxalic acids) did not decrease the biomass of castor bean, despite the fact they reduced the concentration of chlorophyll-a in leaves compared to the control. The Cu concentrations in the roots and shoots significantly increased by 6-106% and 5-148%, respectively, in the LMWOAs treatments so that the total accumulation of Cu by whole plants in all the LMWOAs treatments increased by 21-189% in comparison with the control. The values of the translocation factor (TF) and bio-concentration factor (BCF) of Cu in castor bean also rose following the addition of LMWOAs, indicating that the LMWOAs enhanced the uptake and transportation of Cu. Moreover, the application of LMWOAs did not significantly change the soil pH but significantly increased the activity of soil enzymes (urease, catalase, and alkaline phosphatase). The addition of exogenous LMWOAs increased the available Cu significantly in the soil, thus promoted the phytoextraction efficiency of Cu by castor bean. These results will provide some new insights into the practical use of LMWOAs for the phytoremediation of heavy-metal-contaminated soil employing castor bean., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Castor oil contains approximately 90% ricinoleic acid (RA) which is stored mainly in the form of tri-ricinoleic acid containing triacylglycerols (TAG). Ricinoleate is synthesized from oleate (18:1n-9) esterified to the sn-2 position of phosphatidylcholine (PtdCho) catalyzed by oleoyl-12-hydroxylase. PtdCho-derived diacylglycerol (DAG) is an important substrate pool for TAG synthesis, and the interconversion between PtdCho and DAG has been shown to play a critical role in channeling hydroxy fatty acids (HFA) to TAG. Although phospholipase D (PLD) has been reported to catalyze the hydrolysis of PtdCho to produce phosphatidic acid which can then be converted to DAG, its potential functions in the channeling of RA from PtdCho to DAG and the assembly of RA on TAG is largely unknown. In the present study, 11 PLD genes were identified from the Castor Bean Genome Database. Gene expression analysis indicated that RcPLD9 is expressed at relatively high levels in developing seeds compared to other plant tissues. Sequence and phylogenetic analyses revealed that RcPLD9 is a homolog of Arabidopsis PLDζ2. Overexpression of RcPLD9 in the Arabidopsis CL7 line producing C18-HFA resulted in RA content reductions in the polar lipid fraction (mainly PtdCho) and mono-HFA-TAG, but increased RA content in di-HFA-TAG. Since part of RA in di-HFA-TAG is derived from HFA-DAG, the results indicated that RcPLD9 facilitates the channeling of RA from PtdCho to DAG for its assembly on TAG in developing seeds., (© 2020 AOCS.)

Soil salinity is a major source of abiotic plant stress, adversely affecting plant growth, development and productivity. Although the physiological and molecular mechanisms that underlie plant responses to salt stress are becoming increasingly understood, epigenetic modifications, such as histone methylations and their potential regulation of the transcription of masked genes at the genome level in response to salt stress, remain largely unclear. Castor bean, an important nonedible oil crop, has evolved the capacity to grow under salt stress. Here, based on high-throughput RNA-seq and ChIP-seq data, we systematically investigated changes in genomic transcription and histone methylation using typical histone H3 lysine 4 trimethylation (H3K4me3) and histone H3 tri-methylated lysine 27 (H3K27me3) markers in castor bean leaves subjected to salt stress. The results showed that gain or loss of histone methylation was closely associated with activated or repressed gene expression, though variations in both transcriptome and histone methylation modifications were relatively narrow in response to salt stress. Diverse salt responsive genes and switched histone methylation sites were identified in this study. In particular, we found for the first time that the transcription of the key salt-response regulator RADIALIS-LIKE SANT (RSM1), a MYB-related transcription factor involved in ABA(abscisic acid)-mediated salt stress signaling, was potentially regulated by bivalent H3K4me3-H3K27me3 modifications. Combining phenotypic variations with transcriptional and epigenetic changes, we provide a comprehensive profile for understanding histone modification, genomic transcription and their associations in response to salt stress in plants., (© The Author(s) 2020. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Background: Little is known about the molecular basis of seed size formation in endospermic seed of dicotyledons. The seed of castor bean (Ricinus communis L.) is considered as a model system in seed biology studies because of its persistent endosperms throughout seed development., Results: We compared the size of endosperm and endospermic cells between ZB107 and ZB306 and found that the larger seed size of ZB107 resulted from a higher cell count in the endosperm, which occupy a significant amount of the total seed volume. In addition, fresh weight, dry weight, and protein content of seeds were remarkably higher in ZB107 than in ZB306. Comparative proteomic and transcriptomic analyses were performed between large-seed ZB107 and small-seed ZB306, using isobaric tags for relative and absolute quantification (iTRAQ) and RNA-seq technologies, respectively. A total of 1416 protein species were identified, of which 173 were determined as differentially abundant protein species (DAPs). Additionally, there were 9545 differentially expressed genes (DEGs) between ZB306 and ZB107. Functional analyses revealed that these DAPs and DEGs were mainly involved in cell division and the metabolism of carbohydrates and proteins., Conclusions: These findings suggest that both cell number and storage-component accumulation are critical for the formation of seed size, providing new insight into the potential mechanisms behind seed size formation in endospermic seeds.

Cellular autophagy is a widely-occurring conserved process for turning over damaged organelles or recycling cytoplasmic contents in cells. Although autophagy-related genes ( ATGs ) have been broadly identified from many plants, little is known about the potential function of autophagy in mediating plant growth and development, particularly in recycling cytoplasmic contents during seed development and germination. Castor bean ( Ricinus communis ) is one of the most important inedible oilseed crops. Its mature seed has a persistent and large endosperm with a hard and lignified seed coat, and is considered a model system for studying seed biology. Here, a total of 34 RcATG genes were identified in the castor bean genome and their sequence structures were characterized. The expressional profiles of these RcATGs were examined using RNA-seq and real-time PCR in a variety of tissues. In particular, we found that most RcATGs were significantly up-regulated in the later stage of seed coat development, tightly associated with the lignification of cell wall tissues. During seed germination, the expression patterns of most RcATGs were associated with the decomposition of storage oils. Furthermore, we observed by electron microscopy that the lipid droplets were directly swallowed by the vacuoles, suggesting that autophagy directly participates in mediating the decomposition of lipid droplets via the microlipophagy pathway in germinating castor bean seeds. This study provides novel insights into understanding the potential function of autophagy in mediating seed development and germination., Competing Interests: The authors declare no conflict of interest.

Fertilization is an important agricultural strategy for enhancing the efficiency of phytoremediation in copper (Cu)-contaminated soils. In this study, the effects of nitrogen (N) forms, including ammonium (NH 4 + -N) and nitrate (NO 3 - -N), on the growth, translocation, and accumulation of Cu in the tissues of Ricinus communis L. were investigated in pot and hydroponic experiments. The results demonstrated that higher biomass and N contents in plants were obtained when N was supplied as NO 3 - -N rather than NH 4 + -N. Application of N increased the Cu content in the roots of R. communis, with a higher content after NH 4 + -N (53.10-64.20 mg kg -1 ) than NO 3 - -N (37.62-53.75 mg kg -1 ) treatment. On the contrary, the levels of Cu translocation factors were much higher in NO 3 - -fed plants (0.34-0.45) than in NH 4 + -fed plants (0.28-0.38). The suggested amount of N for fertilizer application is 225 kg hm -2 , which resulted in the highest Cu content in R. communis and optimal plant growth. As the main Cu-binding site, root cell walls accumulated less Cu in plants treated with NH 4 + -N compared with NO 3 - -N. Additionally, NH 4 + -N induced a higher malondialdehyde content and more severe root damage compared with NO 3 - -N. In the leaf, a larger number of black granules, which could be protein and starch grains involved in the detoxification of Cu in R. communis, were present after NH 4 + -N than NO 3 - -N treatment. These results illustrate that N forms are especially important for Cu translocation and accumulation and that immobilization and transformation of Cu in roots were improved more by NH 4 + -N than NO 3 - -N. In conclusion, N fertilizers containing the appropriate forms applied at suitable rates may enhance the biomass and Cu accumulation of R. communis and thereby the remediation efficiency of Cu-contaminated soils.

The physiological and molecular basis of seed size formation is complex, and the development of seed coat (derived from integument cells) might be a critical factor that determines seed size formation for many endospermic seeds. Castor bean ( Ricinus communis L.), a model system of studying seed biology, has large and persistent endosperm with a hard seed coat at maturity. Here, we investigated the potential molecular mechanisms underlying seed size formation in castor bean by comparing the difference between global gene expression within developing seed coat tissues between the large-seed ZB107 and small-seed ZB306. First, we observed the cell size of seed coat and concluded that the large seed coat area of ZB107 resulted from more cell numbers (rather than cell size). Furthermore, we found that the lignin proportion of seed coat was higher in ZB306. An investigation into global gene expression of developing seed coat tissues revealed that 815 genes were up-regulated and 813 were down-regulated in ZB306 relative to ZB107. Interestingly, we found that many genes involved in regulating cell division were up-regulated in ZB107, whereas many genes involved in regulating lignin biosynthesis (including several NAC members, as well as MYB46/83 and MYB58/63) and in mediating programmed cell death (such as CysEP1 and βVPE) were up-regulated in ZB306. Furthermore, the expression patterns of the genes mentioned above indicated that the lignification of seed coat tissues was enhanced and occurred earlier in the developing seeds of ZB306. Taken together, we tentatively proposed a potential scenario for explaining the molecular mechanisms of seed coat governing seed size formation in castor bean by increasing the cell number and delaying the onset of lignification in seed coat tissues in large-seed ZB107. This study not only presents new information for possible modulation of seed coat related genes to improve castor seed yield, but also provides new insights into understanding the molecular basis of seed size formation in endospermic seeds with hard seed coat.

Introduction: Castor (Ricinus communis L.) seeds are valued for their production of oils which can comprise up to 90% hydroxy-fatty acids (ricinoleic acid). Castor oil contains mono-, di- and tri- ricinoleic acid containing triacylglycerols (TAGs). Although the enzymatic synthesis of ricinoleic acid is well described, the differential compartmentalization of these TAG molecular species has remained undefined., Objectives: To examine the distribution of hydroxy fatty acid accumulation within the endosperm and embryo tissues of castor seeds., Methods: Matrix assisted laser desorption/ionization mass spectrometry imaging was used to map the distribution of triacylglycerols in tissue sections of castor seeds. In addition, the endosperm and embryo (cotyledons and embryonic axis) tissues were dissected and extracted for quantitative lipidomics analysis and Illumina-based RNA deep sequencing., Results: This study revealed an unexpected heterogeneous tissue distribution of mono-, di- and tri- hydroxy-triacylglycerols in the embryo and endosperm tissues of castor seeds. Pathway analysis based on transcript abundance suggested that distinct embryo- and endosperm-specific mechanisms may exist for the shuttling of ricinoleic acid away from phosphatidylcholine (PC) and into hydroxy TAG production. The embryo-biased mechanism appears to favor removal of ricinoleic acid from PC through phophatidylcholine: diacylglycerol acyltransferase while the endosperm pathway appears to remove ricinoleic acid from the PC pool by preferences of phospholipase A (PLA 2 α) and/or phosphatidylcholine: diacylglycerol cholinephosphotransferase., Conclusions: Collectively, a combination of lipidomics and transcriptomics analyses revealed previously undefined spatial aspects of hydroxy fatty acid metabolism in castor seeds. These studies underscore a need for tissue-specific studies as a means to better understand the regulation of triacylglycerol accumulation in oilseeds.

Cadmium (Cd) is considered to be the most phytotoxic heavy metal pollutant. The selection of castor bean cultivars with Cd tolerance and the exploration of the physiological mechanisms involved in Cd tolerance are critical steps for improving phytoremediation performance. In this study, a hydroponic experiment was used to investigate variations in Cd transportation, chelation, and subcellular distribution in two different castor bean cultivars, namely JX-22 and ZB-9. Both cultivars had high tolerance index scores, indicating that both cultivars were tolerant to Cd. The findings of the present study indicate that Cd is significantly more mobile in JX-22 than in ZB-9 during xylem and phloem transportation, resulting in the accumulation of Cd in the shoots of JX-22 was 7.67 times that in ZB-9. Subcellular distribution assessment verified that more Cd was bound to the biologically detoxified metal fractions than the metal sensitive fractions in JX-22. The contents of the non-protein thiol pool and glutathione in the leaves were higher in JX-22 than ZB-9 when exposed to Cd. These results indicate that JX-22 has a greater ability to accumulate Cd, and well-coordinated physiological changes in JX-22 afford greater Cd tolerance in comparison to ZB-9 under Cd exposure, indicating that JX-22 is suitable for use in the remediation of Cd-contaminated soils.

The castor bean, Ricinus communis L., is a non-host plant for the large black chafer, Holotrichia parallela Motschulsky (Coleoptera: Scarabaeidae). In laboratory bioassays we found that this plant was no less attractive than the main host plant (peanut, Arachis hypogaea) and three food plant species: velvetleaf (Abutilon theophrasti), the glossy privet (Ligustrum lucidum), and the Siberian elm (Ulmus pumila). In field trapping experiments a Soxhlet extract of castor bean leaves caught more beetles than the optimal sex lure blend [(R)-(-)-linalool and (L)-isoleucine methyl ester blended in a ratio of 1:4], compared at equal doses (500 μl), and laboratory bioassays indicated that a castor bean plant could enhance the attractiveness of different blend ratios of sex lures. Olfactometer bioassays showed that males prefer volatiles emitted from different combinations of castor bean plant extracts and a signaling female over a female alone. In the presence of castor bean plants copulation rates of H. parallela were highest among all test environments both in laboratory bioassays (60%) and in field tests (70%). This study, combined with our previous observation of the feeding behavior of H. parallela adults on castor bean leaves, suggests that castor bean plants may provide an attractive but risky mating site for H. parallela beetles. The enhancement of male mate-location and copulation rate in the presence of castor bean plants can balance its paralytic effects on H. parallela after intake of potential toxins contained in its leaves.

In plants, trehalose 6-phosphate (T6P) is a key signaling metabolite that functions as both a signal and negative feedback regulator of sucrose levels. The mode of action by which T6P senses and regulates sucrose is not fully understood. Here, we demonstrate that the sucrolytic activity of RcSUS1, the dominant sucrose synthase isozyme expressed in developing castor beans, is allosterically inhibited by T6P. The feedback inhibition of SUS by T6P may contribute to the control of sink strength and sucrolytic flux in heterotrophic plant tissues., (© 2018 Federation of European Biochemical Societies.)

While plant oils are an important source of food, plants also produce oils containing specialized fatty acids with chemical and physical properties valued in industry. Ricinoleic acid, a hydroxy fatty acid (HFA) produced in the seed of castor (Ricinus communis), is of particular value, with a wide range of applications. Since castor cultivation is currently successful only in tropical climates, and because castor seed contain the toxin ricin, there are ongoing efforts to develop a temperate crop capable of HFA biosynthesis. In castor, ricinoleic acid is incorporated into triacylglycerol (TAG) which accumulates in the seed lipid droplets. Research in the model plant Arabidopsis (Arabidopsis thaliana) has successfully produced HFA constituting 30% of the total seed oil, but this is far short of the level required to engineer commercially viable crops. Strategies to increase HFA have centered on co-expression of castor TAG biosynthesis enzymes. However, since lipid droplets are the location of neutral lipid storage, manipulating droplets offers an alternative method to increase oil that contains specialized fatty acids. The Arabidopsis Seipin1 protein modulates TAG accumulation by affecting lipid droplet size. Here, we overexpress Seipin1 in a hydroxylase-expressing Arabidopsis line, increasing seed HFA by 62% and proportionally increasing total oil. Increased seed oil was concomitant with a 22% increase in single seed weight and a 69% increase in harvest weight, while seed germination rose by 45%. Because Seipin1 function is unaffected by the structure of the HFA, these results demonstrate a novel strategy that may increase accumulation of many specialized seed oils., (© The Author 2017. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Application of fertilizers to supply appropriate nutrients has become an essential agricultural strategy for enhancing the efficiency of phytoremediation in heavy metal contaminated soils. The present study was conducted to investigate the beneficial effects of three types of phosphate fertilizers (i.e., oxalic acid-activated phosphate rock (APR), Ca(H 2 PO 4 ) 2 , and NaH 2 PO 4 ) in the range of 0-600 mg P kg -1 soil, on castor bean growth, antioxidants [antioxidative enzymes and glutathione (GSH)], and Cu uptake. Results showed that with the addition of phosphorus fertilizers, the dry weight of castor bean and the Cu concentration in roots increased significantly, resulting in increased Cu extraction. The phosphorus concentration in both shoots and roots was increased as compared with the control, and the Ca(H 2 PO 4 ) 2 treatment had the greatest effect. Application of APR, NaH 2 PO 4 , and Ca(H 2 PO 4 ) 2 reduced the malondialdehyde (MDA) content, and the activity of the two antioxidant enzymes superoxide dismustase (SOD, EC 1.15.1.1) and catalase (CAT, EC 1.11.1.6) in the leaves of castor bean. GSH concentration in leaves increased with the increasing levels of phosphorus applied to soil as well as the accumulation of phosphorus in shoots, compared to the control. These results demonstrated that the addition of phosphorus fertilizers can enhance the resistance of castor bean to Cu and increase the Cu extraction efficiency of the plant from contaminated soils.

Ricin is a highly toxic ribosome-inactivating lectin occurring in the seeds of castor bean (Ricinus communis L.). Castor bean grows throughout tropical and sub-tropical regions and is a very important crop due to its high seed content of ricinoleic acid, an unusual fatty acid, which has several industrial applications. However, due to the presence of the toxin, castor bean can cause death after the exposure of animals to low doses of ricin through skin contact, injection, inhalation or oral routes. Aiming to generate a detoxified genotype, we explored the RNAi concept in order to silence the ricin coding genes in the endosperm of castor bean seeds. Results indicated that ricin genes were effectively silenced in genetically modified (GM) plants, and ricin proteins were not detected by ELISA. Hemagglutination activity was not observed with proteins isolated from GM seeds. In addition, we demonstrated that seed proteins from GM plants were not toxic to rat intestine epithelial cells or to Swiss Webster mice. After oil extraction, bio-detoxified castor bean cake, which is very rich in valuable proteins, can be used for animal feeding. Gene silencing would make castor bean cultivation safer for farmers, industrial workers and society.

Ricinus communis L. (castor bean or castor oil plant) was found growing on metal-contaminated sites (4) of peri-urban Greater Hyderabad comprises of erstwhile industrial areas viz Bollaram, Patancheru, Bharatnagar, and Kattedan industrial areas. During 2013-2017, about 60 research papers have appeared focusing the role of castor bean in phytoremediation of co-contaminated soils, co-generation of biomaterials, and environmental cleanup, as bioenergy crop and sustainable development. The present study is focused on its use as a multipurpose phytoremediation crop for phytostabilization and revegetation of waste disposed peri-urban contaminated soils. To determine the plant tolerance level, metal accumulation, chlorophyll, protein, proline, lipid peroxidation, oil content, and soil properties were characterized. It was noticed that the castor plant and soils have high concentration of metals such as cadmium (Cd), lead (Pb), iron (Fe), manganese (Mn), and zinc (Zn). The soils have high phosphorous (P), adequate nitrogen (N), and low concentration of potassium (K). Iron (Fe) concentrations ranged from1672±50.91 to 2166±155.78 mg kg -1 in the soil. The trend of metal accumulation Fe>Zn>Mn>Pb>Cd was found in different plant parts at polluted sites. The translocation of Cd and Pb showed values more than one in industrial areas viz Bollaram, Kattedan, and Bharatnagar indicating the plants resistance to metal toxicity. Chlorophyll and protein content reduced while proline and malondialdehyde increased due to its tolerance level under metal exposure. The content of ricinoleic acid was higher, and the fatty acids composition of polluted areas was almost similar to that of the control area. Thus, R. communis L. can be employed for reclamation of heavy metal contaminated soils.

Response of castor (Ricinus communis L.) to cadmium (Cd) was assessed by a seed-suspending seedbed approach. Length of total radicle was the most sensitive indicator of Cd tolerance among the tested germination and growth characters. The ED50 value for Cd was 11.87 mg L(-1), indicating high Cd tolerance in castor. A pot experiment was conducted by growing 46 varieties of castor under CK (without Cd) and Cd1 (10 mg kg(-1) of Cd) and Cd2 (50 mg kg(-1) of Cd) treatments to investigate genotype variations in growth response and Cd accumulation of castor under different Cd exposures. Castor possessed high Cd accumulation ability; average shoot and root Cd concentrations of the 46 tested varieties were 21.83 and 185.43 mg kg(-1), and 174.99 and 1181.96 mg kg(-1) under Cd1 and Cd2, respectively. Great variation in Cd accumulation was observed among varieties, and Cd concentration of castor was genotype dependent. The correlation between biomass and Cd accumulation was significantly positive, while no significant correlation was observed between Cd concentration and Cd accumulation, which indicated that biomass performance is the dominant factor in determining Cd accumulation ability.

Main Conclusion: The natural OLE-1 high-oleic castor mutant has been characterized, demonstrating that point mutations in the FAH12 gene are responsible for the high-oleic phenotype. The contribution of each mutation was evaluated by heterologous expression in yeast, and lipid studies in developing OLE-1 seeds provided new evidence of unusual fatty acids channeling into TAGs. Ricinus communis L. is a plant of the Euphorbiaceae family well known for producing seeds whose oil has a very high ricinoleic (12-hydroxyoctadecenoic) acid content. Castor oil is considered the only commercially renewable source of hydroxylated fatty acids, which have many applications as chemical reactants. Accordingly, there has been great interest in the field of plant lipid biotechnology to define how ricinoleic acid is synthesized, which could also provide information that might serve to increase the content of other unusual fatty acids in oil crops. Accordingly, we set out to study the biochemistry of castor oil synthesis by characterizing a natural castor bean mutant deficient in ricinoleic acid synthesis (OLE-1). This mutant accumulates high levels of oleic acid and displays remarkable alterations in its seed lipid composition. To identify enzymes that are critical for this phenotype in castor oil, we cloned and sequenced the oleate desaturase (FAD2) and hydroxylase (FAH12) genes from wild-type and OLE-1 castor bean plants and analyzed their expression in different tissues. Heterologous expression in yeast confirmed that three modifications to the OLE-1 FAH12 protein were responsible for its weaker hydroxylase activity. In addition, we studied the expression of the genes involved in this biosynthetic pathway at different developmental stages, as well as that of other genes involved in lipid biosynthesis, both in wild-type and mutant seeds.

In this work, a biorefinery system including castor bean seeds and microalgae is used as a case study to evaluate the integration of second and third generation biorefineries. A biorefinery concept was applied for the combined production of polyol, ethylene-glycol, omega-3 acid, biodiesel, methanol and heat and power from castor bean and microalgae. Castor bean cake and microalgae paste were used to feed a biomass-fired system (BIGCC), where part of CO2 produced in flue gas is captured and employed as substrate for microalgae growth. To evaluate the performance of this biorefinery concept three scenarios based on different levels of mass and energy integration were modeled and assessed from techno-economic and environmental points of view. The scenario with the best economic and environmental performances was the one including full mass integration, full heat integration, and cogeneration scheme., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Aquaporins (AQPs) are a class of integral membrane proteins that facilitate the passive transport of water and other small solutes across biological membranes. Castor bean (Ricinus communis L., Euphobiaceae), an important non-edible oilseed crop, is widely cultivated for industrial, medicinal and cosmetic purposes. Its recently available genome provides an opportunity to analyze specific gene families. In this study, a total of 37 full-length AQP genes were identified from the castor bean genome, which were assigned to five subfamilies, including 10 plasma membrane intrinsic proteins (PIPs), 9 tonoplast intrinsic proteins (TIPs), 8 NOD26-like intrinsic proteins (NIPs), 6 X intrinsic proteins (XIPs) and 4 small basic intrinsic proteins (SIPs) on the basis of sequence similarities. Functional prediction based on the analysis of the aromatic/arginine (ar/R) selectivity filter, Froger's positions and specificity-determining positions (SDPs) showed a remarkable difference in substrate specificity among subfamilies. Homology analysis supported the expression of all 37 RcAQP genes in at least one of examined tissues, e.g., root, leaf, flower, seed and endosperm. Furthermore, global expression profiles with deep transcriptome sequencing data revealed diverse expression patterns among various tissues. The current study presents the first genome-wide analysis of the AQP gene family in castor bean. Results obtained from this study provide valuable information for future functional analysis and utilization.

Biomonitoring of air quality using living organisms is a very interesting approach to environmental impact assessment. Organisms with a vast distribution, such as plants, are widely used for these purposes. The castor bean (Ricinus communis L.) is an oleaginous plant that can potentially be used as a bioindicator plant owing to its rapid growth and large leaves, which have a wide surface area of contact with the air and the pollutants therein. This study investigated the the bioindicator potential of the castor bean by performing several tests. We observed statistically significant differences in the concentrations of chlorophyll a and b in the leaves of plants in polluted areas compared to that in the control group plants, which were located in a pollution-free area. Leaves of plants in the former group had higher peroxidase activity and showed a greater buffering ability than those of plants in the control group. The pKa values obtained via buffering capacity tests, revealed the presence of aminoazobenzene (an industrial dye) in leaves of R. communis. Genotoxicity was evaluated through the comet assay technique and revealed that other than some differences in DNA fragmentation, there is no statistically significant difference in this parameter between places analyzed. Our data indicate that R. communis can be a highly useful biological indicator. Further, we hypothesized that the castor bean can be a potential candidate for phytoremediation owing its physiological buffering capacity when exposed to substantial pollution.

Photoaffinity labeling has been used as a promising approach to detection and isolation of carbohydrate-binding proteins, which are typically characterized by low binding affinity and selectivity. When there are several specific binding proteins, it is desirable that a photoaffinity probe is capable of simultaneously crosslinking them and that the crosslinking yields depend on the relative binding affinities. In this study, we describe the design and synthesis of carbohydrate photoaffinity probes and their ability to capture lectins of different binding affinities., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

We examined the concentrations of metals (Cd, Zn, Cu, Fe and Mn) and potential metal-binding compounds [nicotianamine (NA), thiol compounds and citrate] in xylem and phloem saps from 4-week-old castor bean plants (Ricinus communis) treated with 0 (control), 0.1, 1.0, and 10 μM Cd for 3 weeks. Treatment with 0.1 and 1 μM Cd produced no visible damage, while 10 μM Cd retarded growth. Cadmium concentrations in both saps were higher than those in the culture solution at 0.1 μM, similar at 1.0 μM and lower at 10 μM. Cd at 10 μM reduced Cu and Fe concentrations in both saps. NA concentrations measured by capillary electrophoresis-mass spectrometry (MS) in xylem sap (20 μM) were higher than the Cu concentrations, and those in phloem sap (150 μM) were higher than those of Zn, Fe and Cu combined. Reduced glutathione concentrations differed in xylem and phloem saps (1-2 and 30-150 μM, respectively), but oxidized glutathione concentrations were similar. Phloem sap phytochelatin 2 concentration increased from 0.8 μM in controls to 8 μM in 10 μM Cd. Free citrate was 2-4 μM in xylem sap and 70-100 μM in phloem sap. Total bound forms of Cd in phloem and xylem saps from 1 μM Cd-treated plants were 54 and 8%, respectively. Treatment of phloem sap with proteinaseK reduced high-molecular compounds while increasing fractions of low-molecular Cd-thiol complexes. Zinc-NA, Fe-NA and Cu-NA were identified in the phloem sap fraction of control plants by electrospray ionization time-of-flight MS, and the xylem sap contained Cu-NA., (© 2014 Scandinavian Plant Physiology Society.)

The subcellular localization and chemical forms of copper in castor (Ricinus communis L.) seedlings grown in hydroponic nutrient solution were identified by chemical extraction, transmission electron microscopy, and Fourier transform infrared spectroscopy. The wild castor seeds were harvested from an abandoned copper mine in Tonglu Mountain, Daye City of Hubei Province, China. The results revealed that (1) the seedlings grew naturally in MS liquid medium with 40.00 mg kg(-1) CuSO4, in which the seedling growth rate and biomass index were 0.14 and 1.23, respectively, which were the highest values among all the treatments. The copper content in castor seedlings increased along with elevated CuSO4 concentration in the medium, reaching a maximum value of 16 570.12 mg kg(-1)(DW) when exposed to 60.00 mg L(-1) CuSO4, where 91.31% of the copper was accumulated in roots. (2) The copper existed in various chemical forms in the roots of the castor seedlings. Copper of 67.66% was extracted from the components of cell walls, such as exchangeable acidic polar compounds, cellulose and lignin, protein and pectin, and less concentrated in cell cytoplasm and nuclei. (3) Furthermore, the root cell walls were thickened when the castor seedlings exposed to CuSO4, with a large amount of high-density electron bodies, attached to the thickened cell walls. In the cell walls, most copper was bound to the carboxyl (-COOH) and hydroxyl (-OH) groups of acidic polar compounds, cellulose, hemicellulose, and polysaccharides. The conclusion showed that castor exhibited a strong tolerance to copper, the copper were accumulated mainly in the root cell, the root cell walls of castor were the major location of patience and detoxification in copper stress.

Anoxic conditions should hamper the transport of sugar in the phloem, as this is an active process. The canopy is a carbohydrate source and the roots are carbohydrate sinks. By fumigating the shoot with N2 or flooding the rhizosphere, anoxic conditions in the source or sink, respectively, were induced. Volume flow, velocity, conducting area and stationary water of the phloem were assessed by non-invasive magnetic resonance imaging (MRI) flowmetry. Carbohydrates and δ(13) C in leaves, roots and phloem saps were determined. Following flooding, volume flow and conducting area of the phloem declined and sugar concentrations in leaves and in phloem saps slightly increased. Oligosaccharides appeared in phloem saps and after 3 d, carbon transport was reduced to 77%. Additionally, the xylem flow declined and showed finally no daily rhythm. Anoxia of the shoot resulted within minutes in a reduction of volume flow, conductive area and sucrose in the phloem sap decreased. Sugar transport dropped to below 40% by the end of the N2 treatment. However, volume flow and phloem sap sugar tended to recover during the N2 treatment. Both anoxia treatments hampered sugar transport. The flow velocity remained about constant, although phloem sap sugar concentration changed during treatments. Apparently, stored starch was remobilized under anoxia., (© 2014 John Wiley & Sons Ltd.)

Ricinus communis L. is a bioenergetic crop with high-biomass production and tolerance to cadmium (Cd) and lead (Pb), thus, the plant is a candidate crop for phytoremediation. Pot experiments were performed to study the effects of citric acid in enhancing phytoextraction of Cd/Pb by Ricinus communis L. Citric acid increased Cd and Pb contents in plant shoots in all treatments by about 78% and 18-45%, respectively, at the dosage of 10 mM kg(-1) soil without affecting aboveground biomass production. Addition of citric acid reduced CEC, weakened soil adsorption of heavy metals and activated Cd and Pb in soil solutions. The acid-exchangeable fraction (BCR-1) of Pb remained lower than 7% and significantly increased with citric acid amendment. Respective increases in soil evaluation index induces by 14% and 19% under the Cd1Pb50 and Cd1Pb250 treatments upon addition of citric acid resulted in soil quality improvement. Ricinus communis L. has great potential in citric acid-assisted phytoextraction for Cd and Pb remediation.

Mobile glucose-pesticide conjugates in the phloem are often restricted by decreases in biological activity. However, plants can bioactivate endogenous glucosides, which are assumed as able to bioactivate exogenous conjugates. In this study, four glycosidic bonds (O-, S-, N-, and C-glycosidic bonds) of glucose-pesticide conjugates were designed and synthesized, and then metabolism assays were carried out in vitro and in vivo. Results showed that β-glucosidases played a role in the hydrolysis of O-glycosidic bond conjugates in Ricinus communis L. The liberated aglycons possessed insecticidal activities against Plutella xylostella L. and Spodoptera litura F. These results could help establish methods of circumventing the mutual exclusivity of phloem mobility and biological activity by hydrolyzing endogenous β-glucosidases.

Certainty regarding the degree to which organic molecules exchange oxygen with local water during plant cellulose synthesis (p(ex)) is necessary for cellulose oxygen isotope (δ(18)O(cell))-based applications in environmental and ecological studies. However, the currently accepted notion that p(ex) is a constant of ca. 0.42 appears inconsistent with biochemical theory, which predicts that marked variation may be present in p(ex), in relation to variation in the turnover time (τ) of the carbohydrate pool available for cellulose synthesis. The above prediction was tested in the present study with the analysis of data collected from leaves of Ricinus communis grown in controlled environmental conditions that varied in light intensity and vapour pressure deficit. The results revealed the existence of considerable variation in both p(ex) and τ across plants in the various growth environments. Moreover, despite uncertainties in estimates of the proportion of source water in the synthesis water (p(x)) and of the biochemical fractionation factor (ε(o)), our experiment yielded strong evidence that p(ex) exhibits a significant, positive relationship with τ, consistent with biochemical theory. The observed variation in p(ex) in association with τ has important implications for the interpretation of δ(18)O(cell) data in environmental/ecological studies., (© 2014 John Wiley & Sons Ltd.)

To investigate the effects of Cd on tolerance and antioxidant activities of castor, two different castor (Ricinus communis L.) cultivars (Zibo No. 5 and Zibo No. 8) were used for a hydroponic experiment (0, 1 and 2mg/L Cd) and a pot experiment using Cd-contaminated soil (34mg/kg) with the addition of ethylenedinitrilotetraacetic acid (EDTA). The results indicated that there were significant differences between the two cultivars with respect to Cd uptake in shoots (113-248mg/kg for Zibo No. 5 and 130-288mg/kg Zibo No. 8), biomass tolerance indexes (64.9%-74.6% for Zibo No. 5 and 80.1%-90.9% for Zibo No. 8) in the hydroponic experiment and survival rates (0% for Zibo No. 5 and 100% for Zibo No. 8) determined by the addition of EDTA in the pot experiment, suggesting that Zibo No. 8 has higher tolerance than Zibo No. 5. Moreover, the castor cultivars have low bioconcentration factors (4.80% for Zibo No. 5 and 5.43% for Zibo No. 8) and low translocation factors (<1%). Consequently, Zibo No. 8 can participate in Cd phytostabilization in highly Cd-polluted areas. The results indicated that glutathione (GSH) as a non-enzymatic antioxidant, and antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT) and guaiacol peroxidase (GPX), were cultivar- and dose-dependent. The higher tolerance of Zibo No. 8 compared with Zibo No. 5 can be attributed to the higher GSH levels in the root and higher GPX activity in the leaf., (Copyright © 2014. Published by Elsevier B.V.)

We determined the length, volume, dry biomass, and density in seeds of five castor bean cultivars and verified notable physicochemical trait differences. Seeds were then subjected to different toxic aluminum (Al) concentrations to evaluate germination, relative root elongation, and the role of root apices' rhizosphere mucilage layer. Seeds' physicochemical traits were associated with Al toxicity responses, and the absence of Al in cotyledons near to the embryo was revealed by Al-hematoxylin staining, indicating that Al did not induce significant germination reduction rates between cultivars. However, in the more sensitive cultivar, Al was found around the embryo, contributing to subsequent growth inhibition. After this, to investigate the role of mucilage in Al tolerance, an assay was conducted using NH4Cl to remove root mucilage before or after exposure to different Al concentrations. Sequentially, the roots were stained with hematoxylin and a quantitative analysis of staining intensity was obtained. These results revealed the significant contribution of the mucilage layer to Al toxicity responses in castor bean seedlings. Root growth elongation under Al toxicity confirmed the role of the mucilage layer, which jointly indicated the differential Al tolerance between cultivars and an efficient Al-exclusion mechanism in the tolerant cultivar.

The ability to visualize the movement of glycosyl insecticides contributes to learning their translocation and distribution in plants. In our present work, a novel fluorescent glucose-fipronil conjugate N-[3-cyano-1-[2,6-dichloro-4-(trifluoromethyl)phenyl]-4-[(trifluoromethyl)sulfinyl]-1H-pyrazol-5-yl]-1-(2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-β-D-glucopyranosyl)-1H-1,2,3-triazole-4-methanamine (2-NBDGTF), was obtained via click chemistry. Disk uptake experiments showed that an active carrier-mediated system was involved in the 2-NBDGTF uptake process. Meanwhile, 2-NBDGTF exhibited comparable phloem mobility with GTF in castor bean seedlings. Visualization of 2-NBDGTF uptake and transport experiment showed that this fluorescent glucose-fipronil conjugate could be loaded into sieve tubes after transiting through epidermal cells and mesophyll cells and then translocate from cotyledon to hypocotyl via phloem in castor bean seedlings. The results above determined that it is a promising fluorescence tagging approach for revealing the activities of glycosyl insecticides and 2-NBDGTF is a reasonable and feasible fluorescent surrogate of GTF for tracing the distribution of glucose-fipronil conjugates.

Sieve tube sap was obtained from Tanacetum by aphid stylectomy and from Ricinus after apical bud decapitation. The amino acids in sieve tube sap were analyzed and compared with those from leaves. Arginine and lysine accumulated in the sieve tube sap of Tanacetum more than 10-fold compared to the leaf extracts and they were, together with asparagine and serine, preferably selected into the sieve tube sap, whereas glycine, methionine/tryptophan and γ-amino butyric acid were partially or completely excluded. The two basic amino acids also showed a close covariation in sieve tube sap. The acidic amino acids also grouped together, but antagonistic to the other amino acids. The accumulation ratios between sieve tube sap and leaf extracts were smaller in Ricinus than in Tanacetum. Arginine, histidine, lysine and glutamine were enriched and preferentially loaded into the phloem, together with isoleucine and valine. In contrast, glycine and methionine/tryptophan were partially and γ-amino butyric acid almost completely excluded from sieve tube sap. The covariation analysis grouped arginine together with several neutral amino acids. The acidic amino acids were loaded under competition with neutral amino acids. It is concluded from comparison with the substrate specificities of already characterized plant amino acid transporters, that an AtCAT1-like transporter functions in phloem loading of basic amino acids, whereas a transporter like AtGAT1 is absent in phloem. Although Tanacetum and Ricinus have different minor vein architecture, their phloem loading specificities for amino acids are relatively similar., (© 2014 Scandinavian Plant Physiology Society.)

Background: Compared with major crops, growth and development of Ricinus communis is still poorly understood. A better understanding of the biochemical and physiological aspects of germination and seedling growth is crucial for the breeding of high yielding varieties adapted to various growing environments. In this context, we analysed the effect of temperature on growth of young R. communis seedlings and we measured primary and secondary metabolites in roots and cotyledons. Three genotypes, recommended to small family farms as cash crop, were used in this study., Results: Seedling biomass was strongly affected by the temperature, with the lowest total biomass observed at 20°C. The response in terms of biomass production for the genotype MPA11 was clearly different from the other two genotypes: genotype MPA11 produced heavier seedlings at all temperatures but the root biomass of this genotype decreased with increasing temperature, reaching the lowest value at 35°C. In contrast, root biomass of genotypes MPB01 and IAC80 was not affected by temperature, suggesting that the roots of these genotypes are less sensitive to changes in temperature. In addition, an increasing temperature decreased the root to shoot ratio, which suggests that biomass allocation between below- and above ground parts of the plants was strongly affected by the temperature. Carbohydrate contents were reduced in response to increasing temperature in both roots and cotyledons, whereas amino acids accumulated to higher contents. Our results show that a specific balance between amino acids, carbohydrates and organic acids in the cotyledons and roots seems to be an important trait for faster and more efficient growth of genotype MPA11., Conclusions: An increase in temperature triggers the mobilization of carbohydrates to support the preferred growth of the aerial parts, at the expense of the roots. A shift in the carbon-nitrogen metabolism towards the accumulation of nitrogen-containing compounds seems to be the main biochemical response to support growth at higher temperatures. The biochemical changes observed in response to the increasing temperature provide leads into understanding plant adaptation to harsh environmental conditions, which will be very helpful in developing strategies for R. communis crop improvement research.

In our previous work, a glucose-fipronil (GTF) conjugate at the C-1 position was synthesized via click chemistry and a glucose moiety converted a non-phloem-mobile insecticide fipronil into a moderately phloem-mobile insecticide. In the present paper, fipronil was introduced into the C-2, C-3, C-4, and C-6 positions of glucose via click chemistry to obtain four new conjugates and to evaluate the effects of the different glucose isomers on phloem mobility. The phloem mobility of the four new synthetic conjugates and GTF was tested using the Ricinus seedling system. The results confirmed that conjugation of glucose at different positions has a significant influence on the phloem mobility of GTF conjugates.

Castor bean (Ricinus communis L.) is an important oilseed crop for the rich hydroxylated triacylglycerol (TAG)-ricinoleate which is a raw material with wide applications in industry. Hydroxylated TAG synthesis occurs through complicated pathways among multiple subcellular organelles. Some crucial enzymes have been identified in previous studies. After analyzing the available castor tissue-specific transcriptome sequencing data and comparing the classic pathways in other plants, a possible de novo biosynthesis pathway for the hydroxylated TAG has been revealed. In this study, some other crucial enzymes were ascertained and their expression levels were characterized and pinpointed into the pathways in castor. Several key enzymes were analyzed in terms of structure, biofunction prediction and similarity of expression pattern mechanisms, aiming to give an insight on the better understandings of the molecular knowledge for this oil-rich plant and the crucial enzyme performances in the hydroxylated triacylglycerol-ricinoleate biosynthesis pathways.

Nitrogen isotope signatures in plants might give insights in the metabolism and allocation of nitrogen. To obtain a deeper understanding of the modifications of the nitrogen isotope signatures, we determined δ(15)N in transport saps and in different fractions of leaves, axes and roots during a diel course along the plant axis. The most significant diel variations were observed in xylem and phloem saps where δ(15)N was significantly higher during the day compared with during the night. However in xylem saps, this was observed only in the canopy, but not at the hypocotyl positions. In the canopy, δ(15)N was correlated fairly well between phloem and xylem saps. These variations in δ(15)N in transport saps can be attributed to nitrate reduction in leaves during the photoperiod as well as to (15)N-enriched glutamine acting as transport form of N. δ(15)N of the water soluble fraction of roots and leaves partially affected δ(15)N of phloem and xylems saps. δ(15)N patterns are likely the result of a complex set of interactions and N-fluxes between plant organs. Furthermore, the natural nitrogen isotope abundance in plant tissue is not constant during the diel course - a fact that needs to be taken into account when sampling for isotopic studies., (© 2013 John Wiley & Sons Ltd.)

Oil bodies (OBs) are the intracellular particles derived from oilseeds. These OBs store lipids as a carbon resource, and have been exploited for a variety of industrial applications including biofuels. Oleosin and caleosin are the common OB structural proteins which are enabling biotechnological enhancement of oil content and OB-based pharmaceutical formations via stabilizing OBs. Although the draft whole genome sequence information for Ricinus communis L. (castor bean) and Linum usitatissimum L. (flax), important oil seed plants, is available in public database, OB-structural proteins in these plants are poorly indentified. Therefore, in this study, we performed a comprehensive bioinformatic analysis including analysis of the genome sequence, conserved domains and phylogenetic relationships to identify OB structural proteins in castor bean and flax genomes. Using comprehensive analysis, we have identified 6 and 15 OB-structural proteins from castor bean and flax, respectively. A complete overview of this gene family in castor bean and flax is presented, including the gene structures, phylogeny and conserved motifs, resulting in the presence of central hydrophobic regions with proline knot motif, providing an evolutionary proof that this central hydrophobic region had evolved from duplications in the primitive eukaryotes. In addition, expression analysis of L-oleosin and caleosin genes using quantitative real-time PCR demonstrated that seed contained their maximum expression, except that RcCLO-1 expressed maximum in cotyledon. Thus, our comparative genomics analysis of oleosin and caleosin genes and their putatively encoded proteins in two non-model plant species provides insights into the prospective usage of gene resources for improving OB-stability., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Bioenergy production combined with phytoremediation has been suggested to help in solving two critical world problems: the gradual reduction of fossil fuels and soil contamination. The aim of this research was to investigate the potential for the use of Ricinus communis L. (castor oil plant) as an energy crop and plant species to remediate metal-polluted sites. This study was performed in mine tailings containing high concentrations of Cu, Zn, Mn, Pb and Cd. Physico-chemical characterization, total, DTPA-extractable and water-soluble metals in rhizospheric tailings heap samples were carried. Metal concentrations in plant tissues and translocation factors (TFs) were also determined. The Ricinus seed-oil content was high between 41 and 64%, seeds from San Francisco site 6 had the highest oil content, while these from site 7 had the lowest. No trend between oil yield vs seed origin site was observed. Seed-oil content was negatively correlated with root concentration of Cu, Zn, Pb and Cd, but no correlation was observed with the extractable-metals. According to its shoot metal concentrations and TFs, castor bean is not a metal accumulator plant. This primary colonizing plant is well suited to cope with the local toxic conditions and can be useful for the stabilization of these residues, and for then decreasing metal bioavailability, dispersion and human health risks on these barren tailings heaps and in the surrounding area. Our work is the first report regarding combined oil production and a phytostabilization role for Ricinus plants in metal mine tailings and may give a new value to suitable metal-polluted areas., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

A pot experiment was conducted to investigate the potential for phytoextraction of heavy metals and rhizoremediation of polycyclic aromatic hydrocarbons (PAHs) in co-contaminated soil by co-planting a cadmium/zinc (Cd/Zn) hyperaccumulator and lead (Pb) accumulator Sedum alfredii with ryegrass (Lolium perenne) or castor (Ricinus communis). Co-planting with castor decreased the shoot biomass of S. alfredii as compared to that in monoculture. Cadmium concentration in S. alfredii shoot significantly decreased when grown with ryegrass or castor as compared to that in monoculture. However, no reduction of Zn or Pb concentration in S. alfredii shoot was detected in co-planting treatments. Total removal of either Cd, Zn, or Pb by plants was similar across S. alfredii monoculture or co-planting with ryegrass or castor, except enhanced Pb removal in S. alfredii and ryegrass co-planting treatment. Co-planting of S. alfredii with ryegrass or castor significantly enhanced the pyrene and anthracene dissipation as compared to that in the bare soil or S. alfredii monoculture. This appears to be due to the increased soil microbial population and activities in both co-planting treatments. Co-planting of S. alfredii with ryegrass or castor provides a promising strategy to mitigate both metal and PAH contaminants from co-contaminated soils.

We have previously reported that Ricinus communis (castor) is more tolerant to soil cadmium (Cd) and more efficient for Cd phytoremediation than Brassica juncea (Indian mustard) (Bauddh and Singh, 2012). In the present study, R. communis was found more tolerant to salinity and drought in presence of Cd and removed more Cd in a given time than Indian mustard. R. communis produced 23 and twelve folds higher biomass in terms of fresh weight and dry weight, respectively than that in B. juncea during three months when grown in Cd contaminated soil in presence of 100mM NaCl salinity and ten day water withdrawal based drought at 90 day after sowing (DAS). Castor plants showed stronger self-protection ability in form of proline bioaccumulation (r(2)=0.949) than Indian mustard (r(2)=0.932), whereas a lower r(2) for malondialdehyde (MDA) and total soluble protein in R. communis (r(2)=0.914 and r(2)=0.915, respectively) than that of B. juncea (r(2)=0.947 and r(2)=0.927, respectively) indicated a greater damage to cell membrane in Indian mustard during the multiple stress conditions. Though, the amount of Cd accumulated in the roots and shoots of Indian mustard was higher as per unit biomass than that in castor, total removal of the metal from soil was much higher in castor on per plant basis in the same period in presence of the stresses. R. communis accumulated about seventeen and 1.5 fold higher Cd in their roots and shoots, respectively than that of B. juncea in 90 DAS under the multiple stresses. Salinity alone enhanced Cd uptake, whereas drought stress reduced its uptake in both the plants., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Reports about diurnal changes of assimilates in phloem sap are controversial. We determined the diurnal changes of sucrose and amino acid concentrations and fluxes in exudates from cut aphid stylets on tansy leaves (Tanacetum vulgare), and sucrose, amino acid and K(+) concentrations and fluxes in bleeding sap of castor bean pedicel (Ricinus communis). Approximately half of the tansy sieve tubes exhibited a diurnal cycle of sucrose concentrations and fluxes in phloem sap. Data from many tansy plants indicated an increased sucrose flux in the phloem during daytime in case of low N-nutrition, not at high N-nutrition. The sucrose concentration in phloem sap of young Ricinus plants changed marginally between day and night, whereas the sucrose flux increased 1.5-fold during daytime (but not in old Ricinus plants). The amino acid concentrations and fluxes in tansy sieve tubes exhibited a similar diurnal cycle as the sucrose concentrations and fluxes, including their dependence on N-nutrition. The amino acid fluxes, but not the concentrations, in phloem sap of Ricinus were higher at daytime. The sucrose/amino acid ratio showed no diurnal cycle neither in tansy nor in Ricinus. The K(+)-concentrations in phloem sap of Ricinus, but not the K(+) fluxes, decreased slightly during daytime and the sucrose/K(+)-ratio increased. In conclusion, a diurnal cycle was observed in sucrose, amino acid and K(+) fluxes, but not necessarily in concentrations of these assimilates. Because of the large variations between different sieve tubes and different plants, the nutrient delivery to sink tissues is not homeostatic over time.

Some compounds containing glucose are absorbed via the monosaccharide transporters of the plasma membrane. A glucose-fipronil conjugate, N-[3-cyano-1-[2,6-dichloro-4-(trifluoromethyl)phenyl]-4-[(trifluoromethyl)sulfinyl]-1H-pyrazol-5-yl]-1-(β-d-glucopyranosyl)-1H-1,2,3-triazole-4-methanamine (GTF), has been synthesized in our previous work. GTF exhibits moderate phloem mobility in Ricinus communis. In the current paper, we demonstrate that the uptake of GTF by Ricinus seedling cotyledon discs is partly mediated by an active carrier system (K(m)1 = 0.17 mM; V(max)1 = 2.2 nmol cm(-2) h(-1)). Four compounds [d-glucose, sucrose, phloridzin, and carbonyl cyanide m-chlorophenylhydrazone (CCCP)] were examined for their effect on GTF uptake. Phloridzin as well as CCCP markedly inhibit GTF uptake, and d-glucose weakly competes with it. The phloem transport of GTF in Ricinus seedlings is found to involve an active carrier-mediated mechanism that effectively contributes to the GTF phloem loading. The results prove that adding a glucose core is a reasonable and feasible approach to confer phloem mobility to fipronil by utilizing plant monosaccharide transporters.

This study evaluated Cd and Pb accumulation by castor bean (Ricinus communis cv. Guarany) plants grown in nutrient solution, aiming to assess the plant's ability and tolerance to grow in Cd- and Pb-contaminated solutions for phytoremediation purposes. The plants were grown in individual pots containing Hoagland and Arnon's nutrient solution with increasing concentrations of Cd and Pb. The production of root and shoot dry matter and their contents of Cd, Pb, Ca, Mg, Cu, Fe, Mn, and Zn were evaluated in order to calculate the translocation and bioaccumulation factors, as well as toxicity of Cd and Pb. Cadmium caused severe symptoms of phytotoxicity in the plant's root and shoot, but no adverse effect was observed for Pb. Castor bean is an appropriate plant to be used as indicator plant for Cd and tolerante for Pb in contaminated solution and it can be potentially used for phytoremediation of contaminated areas.