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Publikation

Cheng, Y.; Zhao, H.; Cui, L.; Hussain, H.; Nadolnik, L.; Zhang, Z.; Zhao, Y.; Qin, X.; Li, J.; Park, J. H.; Wang, D.; Ultrasonic-assisted extraction of flavonoids from peanut leave and stem using deep eutectic solvents and its molecular mechanism Food Chem. 434, 137497, (2024) DOI: 10.1016/j.foodchem.2023.137497

Natural bioactive compounds extracted from agricultural by-products have received considerable attentions. Twenty-two kinds of deep eutectic solvents (DESs) with ultrasonic were screened to extract flavonoids from peanut leave and stem. ChCl-acetic acid (ChCl-Aa) with 1:2 M ratio resulted in more effective extraction of flavonoids compared to other solvents The best extraction conditions were found to be at a 27% water content in DES/H2O, for 43 min with 31:1 g/mL liquid/solid ratio, giving 2.980 mg/g dw of flavonoids through the response surface method. SEM showed that ChCl-Aa had a certain dissociation impact on the sample matrix, while 1H NMR analysis revealed the formation of hydrogen bonds between daidzein and ChCl-Aa. Changes in the H–bond length and number were observed by the B3LYP/6-31G (d,p) level of theory to confirm the experimental spectra. This study reveals that DESs are efficient for obtaining value-added products and could applied to other natural products.
Publikation

Xie, L.; Yan, H.; Han, L.; Cui, L.; Hussain, H.; Feng, Q.; Zhao, Y.; Zhang, Z.; Li, J.; Aziz, S.; He, J.; Wang, D.; Structural characterization and anti-inflammatory activity of neutral polysaccharides from American ginseng Int. J. Biol. Macromol. 248, 125586, (2023) DOI: 10.1016/j.ijbiomac.2023.125586

American ginseng, a precious classic herbal medicine, is used extensively in China for life prolongation purpose. This study aimed to elucidate the structure and anti-inflammatory activity of a neutral polysaccharide isolated from American ginseng (AGP-A). Nuclear magnetic resonance in conjunction with gas chromatography–mass spectrometry were used to analyze AGP-A\'s structure, whereas Raw264.7 cell and zebrafish models were employed to assess its anti-inflammatory activity. According to the results, AGP-A has a molecular weight of 5561 Da and is primarily consisted of glucose. Additionally, linear α-(1 → 4)-glucans with α-D-Glcp-(1 → 6)-α-Glcp-(1→ residues linked to the backbone at C-6 formed the backbone of AGP-A. Furthermore, AGP-A significantly decreased pro-inflammatory cytokines (IL-1β, IL-6 and TNF-α) in Raw264.7 cell model. AGP-A in zebrafish model significantly lower the massive recruitment of neutrophils to the neuromast of the caudal lateral line. Inflammation may be relieved by the AGP-A component in American ginseng based on these results. In conclusion, our study shows the structural characterization, remarkable anti-inflammatory properties of AGP-A and its potential curative efficacy as a safe, valid natural anti-inflammatory medicine.
Publikation

Sun, X.; Xu, L.; Yan, H.; Li, P.; Hussain, H.; Liu, J.; Zhang, J.; Wang, D.; Isolation and purification of high polar glycosides from aerial parts of Gynostemma pentaphyllum (Thunb.) Makino by linear gradient counter‐current chromatography coupled with inner‐recycling mode J Sep Sci 46, 2300238, (2023) DOI: 10.1002/jssc.202300238

Gynostemma pentaphyllum (Thunb.) Makino represents the popular health food and supplemental product with broad pharmacological activities. The highly polar glycosides, including flavonoids and saponins, are major effective active components that contain diverse sugar positions and quantities, which result in diverse chemical polarities, making it challenging to separate and isolate these components. The present work described the rapid and efficient linear gradient counter‐current chromatography to preparatively separate glycosides from aboveground parts of G. pentaphyllum. Besides, the ethyl acetate and n‐butanol binary mobile phases were achieved through adjusting associated proportions. Six glycosides, including quercetin‐3‐O‐neohesperidoside (1), kaempferol‐3‐O‐robinobioside (2), kaempferol‐3‐O‐neohesperidoside (3), gypenoside LVI (4), ginsenoside Rb3 (5), and gypenoside XLVI (6), were isolated at the purities greater than 98%. Moreover, electrospray ionization mass spectrometry and nuclear magnetic resonance tandem mass spectrometry were conducted for structural identification. According to our findings, the established linear gradient counter‐current chromatography was an efficient approach to separate the highly polar glycosides from aboveground parts of G. pentaphyllum. Our proposed strategy can be used to separate active compounds from other complex natural products.
Publikation

Dahiya, P.; Bürstenbinder, K.; The making of a ring: Assembly and regulation of microtubule-associated proteins during preprophase band formation and division plane set-up Curr. Opin. Plant Biol. 73, 102366, (2023) DOI: 10.1016/j.pbi.2023.102366

The preprophase band (PPB) is a transient cytokinetic structure that marks the future division plane at the onset of mitosis. The PPB forms a dense cortical ring of mainly microtubules, actin filaments, endoplasmic reticulum, and associated proteins that encircles the nucleus of mitotic cells. After PPB disassembly, the positional information is preserved by the cortical division zone (CDZ). The formation of the PPB and its contribution to timely CDZ set-up involves activities of functionally distinct microtubule-associated proteins (MAPs) that interact physically and genetically to support robust division plane orientation in plants. Recent studies identified two types of plant-specific MAPs as key regulators of PPB formation, the TON1 RECRUITMENT MOTIF (TRM) and IQ67 DOMAIN (IQD) families. Both families share hallmarks of disordered scaffold proteins. Interactions of IQDs and TRMs with multiple binding partners, including the microtubule severing KATANIN1, may provide a molecular framework to coordinate PPB formation, maturation, and disassembly.
Publikation

Ning, F.; Zhu, H.; Yan, H.; Liu, J.; Aziz, S.; Hussain, H.; Song, X.; Xie, L.; Meng, Z.; Cao, G.; Wang, D.; Separation and purification of quinolyridine alkaloids from seeds of Thermopsis lanceolata R. Br. by conventional and pH‐zone‐refining counter‐current chromatography J Sep Sci 46, 2300053, (2023) DOI: 10.1002/jssc.202300053

In this work, the preparative separation of quinolyridine alkaloids from seeds of T. lanceolata by conventional and pH-zone-refining counter-current chromatography. Traditional counter-current chromatography separation was performed by a flow-rate changing strategy with a solvent system of ethyl acetate-n-butanol-water (1:9:10, v/v) and 200 mg sample loading. Meanwhile, the pH-zone-refining mode was adopted for separating 2.0 g crude alkaloid extracts with the chloroform-methanol-water (4:3:3, v/v) solvent system using the stationary and mobile phases of 40 mM hydrochloric acid and 10 mM triethylamine. Finally, six compounds, including N-formylcytisine (two conformers) (1), N-acetycytisine (two conformers) (2), (-)-cytisine (3), 13-β-hydroxylthermopsine (4), N-methylcytisine (5), and thermopsine (6) were successfully obtained in the two counter-current chromatography modes with the purities over 96.5%. Moreover, we adopted nuclear magnetic resonance and mass spectrometry for structural characterization. Based on the obtained findings, the pH-zone-refining mode was the efficient method to separate quinolyridine alkaloids relative to the traditional mode.
Publikation

Nietzschmann, L.; Smolka, U.; Perino, E. H. B.; Gorzolka, K.; Stamm, G.; Marillonnet, S.; Bürstenbinder, K.; Rosahl, S.; The secreted PAMP-induced peptide StPIP1_1 activates immune responses in potato Sci. Rep. 13, 20534, (2023) DOI: 10.1038/s41598-023-47648-x

Treatment of potato plants with the pathogen-associated molecular pattern Pep-13 leads to the activation of more than 1200 genes. One of these, StPIP1_1, encodes a protein of 76 amino acids with sequence homology to PAMP-induced secreted peptides (PIPs) from Arabidopsis thaliana. Expression of StPIP1_1 is also induced in response to infection with Phytophthora infestans, the causal agent of late blight disease. Apoplastic localization of StPIP1_1-mCherry fusion proteins is dependent on the presence of the predicted signal peptide. A synthetic peptide corresponding to the last 13 amino acids of StPIP1_1 elicits the expression of the StPIP1_1 gene itself, as well as that of pathogenesis related genes. The oxidative burst induced by exogenously applied StPIP1_1 peptide in potato leaf disks is dependent on functional StSERK3A/B, suggesting that StPIP1_1 perception occurs via a receptor complex involving the co-receptor StSERK3A/B. Moreover, StPIP1_1 induces expression of FRK1 in Arabidopsis in an RLK7-dependent manner. Expression of an RLK from potato with high sequence homology to AtRLK7 is induced by StPIP1_1, by Pep-13 and in response to infection with P. infestans. These observations are consistent with the hypothesis that, upon secretion, StPIP1_1 acts as an endogenous peptide required for amplification of the defense response.
Publikation

Bürstenbinder, K.; Schwarzerová, K.; European Plant Cytoskeletal Club meeting: A vital platform for advancing plant cytoskeleton research Cytoskeleton 80, 397-399, (2023) DOI: 10.1002/cm.21780

This contribution reports on a meeting of plant cytoskeleton scientists-the European Plant Cytoskeletal Club 2023 conference.
Publikation

Bao, Z.; Guo, Y.; Deng, Y.; Zang, J.; Zhang, J.; Deng, Y.; Ouyang, B.; Qu, X.; Bürstenbinder, K.; Wang, P.; Microtubule-associated protein SlMAP70 interacts with IQ67-domain protein SlIQD21a to regulate fruit shape in tomato Plant Cell 35, 4266-4283, (2023) DOI: 10.1093/plcell/koad231

Tomato (Solanum lycopersicum) fruit shape is related to microtubule organization and the activity of microtubule-associated proteins (MAPs). However, insights into the mechanism of fruit shape formation from a cell biology perspective remain limited. Analysis of the tissue expression profiles of different microtubule regulators revealed that functionally distinct classes of MAPs, including members of the plant-specific MICROTUBULE-ASSOCIATED PROTEIN 70 (MAP70) and IQ67 DOMAIN (IQD, also named SUN in tomato) families, are differentially expressed during fruit development. SlMAP70-1–3 and SlIQD21a are highly expressed during fruit initiation, which relates to the dramatic microtubule pattern rearrangements throughout this developmental stage of tomato fruits. Transgenic tomato lines overexpressing SlMAP70-1 or SlIQD21a produced elongated fruits with reduced cell circularity and microtubule anisotropy, while their loss-of-function mutants showed the opposite phenotype, harboring flatter fruits. Fruits were further elongated in plants coexpressing both SlMAP70-1 and SlIQD21a. We demonstrated that SlMAP70s and SlIQD21a physically interact and that the elongated fruit phenotype is likely due to microtubule stabilization induced by the SlMAP70–SlIQD21a interaction. Together, our results identify SlMAP70 proteins and SlIQD21a as important regulators of fruit elongation and demonstrate that manipulating microtubule function during early fruit development provides an effective approach to alter fruit shape.
Bücher und Buchkapitel

Klemm, S.; Buhl, J.; Möller, B.; Bürstenbinder, K.; Quantitative analysis of microtubule organization in leaf epidermis pavement cells (Hussey, P.J., Wang, P.). The Plant Cytoskeleton 2604, 43-61, (2023) ISBN: 978-1-0716-2866-9 DOI: 10.1007/978-1-0716-2867-6_4

Leaf epidermis pavement cells form highly complex shapes with interlocking lobes and necks at their anticlinal walls. The microtubule cytoskeleton plays essential roles in pavement cell morphogenesis, in particular at necks. Vice versa, shape generates stress patterns that regulate microtubule organization. Genetic or pharmacological perturbations that affect pavement cell shape often affect microtubule organization. Pavement cell shape and microtubule organization are therefore closely interconnected. Here, we present commonly used approaches for the quantitative analysis of pavement cell shape characteristics and of microtubule organization. In combination with ablation experiments, these methods can be applied to investigate how different genotypes (or treatments) affect the organization and stress responsiveness of the microtubule cytoskeleton.
Preprints

Bao, Z.; Guo, Y.; Deng, Y.; Zang, J.; Zhang, J.; Ouyang, B.; Qu, X.; Bürstenbinder, K.; Wang, P.; The microtubule-associated protein SlMAP70 interacts with SlIQD21 and regulates fruit shape formation in tomato (2022) DOI: 10.1101/2022.08.08.503161

The shape of tomato fruits is closely correlated to microtubule organization and the activity of microtubule associated proteins (MAP), but insights into the mechanism from a cell biology perspective are still largely elusive. Analysis of tissue expression profiles of different microtubule regulators revealed that functionally distinct classes of MAPs are highly expressed during fruit development. Among these, several members of the plant-specific MAP70 family are preferably expressed at the initiation stage of fruit development. Transgenic tomato lines overexpressing SlMAP70 produced elongated fruits that show reduced cell circularity and microtubule anisotropy, while SlMAP70 loss-of-function mutant showed an opposite effect with flatter fruits. Microtubule anisotropy of fruit endodermis cells exhibited dramatic rearrangement during tomato fruit development, and SlMAP70-1 is likely implicated in cortical microtubule organization and fruit elongation throughout this stage by interacting with SUN10/SlIQD21a. The expression of SlMAP70 (or co-expression of SlMAP70 and SUN10/SlIQD21a) induces microtubule stabilization and prevents its dynamic rearrangement, both activities are essential for fruit shape establishment after anthesis. Together, our results identify SlMAP70 as a novel regulator of fruit elongation, and demonstrate that manipulating microtubule stability and organization at the early fruit developmental stage has a strong impact on fruit shape.
  • Die Leibniz-Gemeinschaft
  • Digitalisierung in der Landwirtschaft
  • Martin-Luther Universität Halle
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