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1
Coelho, Marco A., André Rosa, Nádia Rodrigues, Álvaro Fonseca, and Paula Gonçalves. "Identification of Mating Type Genes in the Bipolar Basidiomycetous Yeast Rhodosporidium toruloides: First Insight into the MAT Locus Structure of the Sporidiobolales." Eukaryotic Cell 7, no. 6 (April 11, 2008): 1053–61. http://dx.doi.org/10.1128/ec.00025-08.
Повний текст джерелаАнотація:
ABSTRACT Rhodosporidium toruloides is a heterothallic, bipolar, red yeast that belongs to the Sporidiobolales, an order within a major lineage of basidiomycetes, the Pucciniomycotina. In contrast to other basidiomycetes, considerably less is known about the nature of the mating type (MAT) loci that control sexual reproduction in this lineage. Three genes (RHA1, RHA2, and RHA3) encoding precursors of the MAT A1 pheromone (rhodotorucine A) were previously identified and formed the basis for a genome walking approach that led to the identification of additional MAT genes in complementary mating strains of R. toruloides. Two mating type-specific alleles encoding a p21-activated kinase (PAK; Ste20 homolog) were found between the RHA2 and RHA3 genes, and identification in MAT A2 strains of a gene encoding a presumptive pheromone precursor enabled prediction of the structure of rhodotorucine a. In addition, a putative pheromone receptor gene (STE3 homolog) was identified upstream of RHA1. Analyses of genomic data from two closely related species, Sporobolomyces roseus and Sporidiobolus salmonicolor, identified syntenic regions that contain homologs of all the above-mentioned genes. Notably, six novel pheromone precursor genes were uncovered, which encoded, similarly to the RHA genes, multiple tandem copies of the peptide moiety. This suggests that this structure, which is unique among fungal lipopeptide pheromones, seems to be prevalent in red yeasts. Species comparisons provided evidence for a large, multigenic MAT locus structure in the Sporidiobolales, but no putative homeodomain transcription factor genes (which are present in all basidiomycetous MAT loci characterized thus far) could be found in any of the three species in the vicinity of the MAT genes identified.
2
Rivers, Damien M. R., and Ivan J. Oresnik. "The Sugar Kinase That Is Necessary for the Catabolism of Rhamnose in Rhizobium leguminosarum Directly Interacts with the ABC Transporter Necessary for Rhamnose Transport." Journal of Bacteriology 197, no. 24 (September 28, 2015): 3812–21. http://dx.doi.org/10.1128/jb.00510-15.
Повний текст джерелаАнотація:
ABSTRACTRhamnose catabolism inRhizobium leguminosarumwas found to be necessary for the ability of the organism to compete for nodule occupancy. Characterization of the locus necessary for the catabolism of rhamnose showed that the transport of rhamnose was dependent upon a carbohydrate uptake transporter 2 (CUT2) ABC transporter encoded byrhaSTPQand on the presence of RhaK, a protein known to have sugar kinase activity. A linker-scanning mutagenesis analysis ofrhaKshowed that the kinase and transport activities of RhaK could be separated genetically. More specifically, two pentapeptide insertions defined by the allelesrhaK72andrhaK73were able to uncouple the transport and kinase activities of RhaK, such that the kinase activity was retained, but cells carrying these alleles did not have measurable rhamnose transport rates. These linker-scanning alleles were localized to the C terminus and N terminus of RhaK, respectively. Taken together, the data led to the hypothesis that RhaK might interact either directly or indirectly with the ABC transporter defined byrhaSTPQ. In this work, we show that both N- and C-terminal fragments of RhaK are capable of interacting with the N-terminal fragment of the ABC protein RhaT using a 2-hybrid system. Moreover, if RhaK fragments carrying either therhaK72orrhaK73allele were used, this interaction was abolished. Phylogenetic and bioinformatic analysis of the RhaK fragments suggested that a conserved region in the N terminus of RhaK may represent a putative binding domain. Alanine-scanning mutagenesis of this region followed by 2-hybrid analysis revealed that a substitution of any of the conserved residues greatly affected the interaction between RhaT and RhaK fragments, suggesting that the sugar kinase RhaK and the ABC protein RhaT interact directly.IMPORTANCEABC transporters involved in the transport of carbohydrates help define the overall physiological fitness of bacteria. The two largest groups of transporters are thecarbohydrateuptaketransporter classes 1 and 2 (CUT1 and CUT2, respectively). This work provides the first evidence that a kinase that is necessary for the catabolism of a sugar can directly interact with a domain from the ABC protein that is necessary for its transport.
3
Thavornwatanayong, Thongthai, Simin Zheng, Sabine Jean Guillaume, and Bao Q. Vuong. "The DEAH-box helicase RHAU regulates immunoglobulin class switch recombination." Journal of Immunology 208, no. 1_Supplement (May 1, 2022): 112.08. http://dx.doi.org/10.4049/jimmunol.208.supp.112.08.
Повний текст джерелаАнотація:
Abstract During a humoral immune response, B cells produce alternative immunoglobulin isotypes (IgG, IgA, IgE) through class switch recombination (CSR). Initiation of CSR requires AID-mediated deamination of switch (S) regions in the immunoglobulin heavy chain (IgH) locus. Although G-quadruplex(G4)-forming S region RNAs bind AID and localize AID to S region DNA sequences, the molecular mechanism that regulates the transition of AID binding from RNA to DNA remains uncharacterized. Highly stable G4 structures necessitate helicases to unwind the G-G hydrogen bonds, which in turn may permit the base-pairing of the S transcript to the complementary DNA sequence in the IgH locus during CSR. An S region RNA pull-down assay identified the RNA Helicase associated with AU-rich element (RHAU), a DEAH-box RNA helicase, as an S transcript binding protein. To examine the role of RHAU in CSR, we genetically deleted a floxed RHAU allele in B cells of mice using CD23-Cre. Conditional deletion of RHAU in B cells reduced CSR in vivo and in vitro to approximately 50% of wild-type controls. This data suggests that RHAU plays an important role in CSR. We hypothesize that RHAU unwinds G4 in S transcripts to facilitate the handoff of AID from G4-RNA to S region DNA during CSR. Supported by The National Cancer Institute (2U54CA132378) and The National Institute of General Medical Sciences (1SC1GM132035-01)
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Richardson, Jason S., and Ivan J. Oresnik. "l-Rhamnose Transport Is Sugar Kinase (RhaK) Dependent in Rhizobium leguminosarum bv. trifolii." Journal of Bacteriology 189, no. 23 (September 21, 2007): 8437–46. http://dx.doi.org/10.1128/jb.01032-07.
Повний текст джерелаАнотація:
ABSTRACT Strains of Rhizobium leguminosarum which are unable to catabolize l-rhamnose, a methyl-pentose sugar, are compromised in the ability to compete for nodule occupancy versus wild-type strains. Previous characterization of the 11-kb region necessary for the utilization of rhamnose identified a locus carrying catabolic genes and genes encoding the components of an ABC transporter. Genetic evidence suggested that the putative kinase RhaK carried out the first step in the catabolism of rhamnose. Characterization of this kinase led to the observation that strains carrying rhamnose kinase mutations were unable to transport rhamnose into the cell. The absence of a functional rhamnose kinase did not stop the transcription and translation of the ABC transporter components. By developing an in vitro assay for RhaK activity, we have been able to show that (i) RhaK activity is consistent with RhaK phosphorylating rhamnose and (ii) biochemical activity of RhaK is necessary for rhamnose transport.
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Huang, Minghui, Ruifeng Qin, Chunjie Li, Mingze Wang, Ye Jiang, Jinyao Yu, Doudou Chang, et al. "Response of soybean genotypes from Northeast China to Heterodera glycines races 4 and 5, and characterisation of rhg1 and Rhg4 genes for soybean resistance." Nematology 24, no. 3 (October 7, 2021): 333–45. http://dx.doi.org/10.1163/15685411-bja10134.
Повний текст джерелаАнотація:
Summary Soybean cyst nematode (SCN, Heterodera glycines) is a devastating plant-parasitic nematode worldwide. Two SCN races, race 4 (HG Type 1.2.3.5.6.7) and race 5 (HG Type 2.5.7), with increased virulence were previously identified in Northeast China. To obtain new resistance sources to these SCN populations, the response of 62 genotypes, including 51 local varieties and breeding lines, and 11 indicator lines for SCN race and HG Type identification, were evaluated. Four new primers in the regions of two loci of GmSHMT08 (Rhg4) and GmSNAP18 (rhg1) were designed for PCR amplification and subsequent sequencing to characterise haplotypes instead of genome resequencing. Results indicated three haplotypes among 51 local genotypes; there were 26 lines in Haplotype I carrying both the rhg1-a and Rhg4-a resistant loci as in ‘Peking’, 13 lines in Haplotype II containing only the resistant rhg1-a locus but Rhg4-b susceptible loci, and 12 lines in Haplotype III with rhg1-c and Rhg4-b susceptible loci. Interestingly, there was no ‘PI 88788’-type resistance identified in Northeast China, although it accounts for 90% of sources in the USA. Two local breeding lines in Haplotype I displayed resistance to both SCN races. The resistance lines carried higher copy number (>1) of the tandem duplication at the rhg1 locus compared with susceptible lines (⩽1). The combination of the two microsatellite markers, Sat_162 on Chr 8 and 590 on Chr 18, distinguished the three haplotypes and predicted the resistance/susceptibility for SCN race 5. The knowledge of the phenotypes and molecular characteristics of 51 local breeding lines in Northeast China will accelerate the utilisation of sources for broad-based SCN resistance and marker-assisted selection.
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Richards, Emma, Laura Bouché, Maria Panico, Ana Arbeloa, Evgeny Vinogradov, Howard Morris, Brendan Wren, Susan M. Logan, Anne Dell, and Neil F. Fairweather. "The S-layer protein of a Clostridium difficile SLCT-11 strain displays a complex glycan required for normal cell growth and morphology." Journal of Biological Chemistry 293, no. 47 (October 1, 2018): 18123–37. http://dx.doi.org/10.1074/jbc.ra118.004530.
Повний текст джерелаАнотація:
Clostridium difficile is a bacterial pathogen that causes major health challenges worldwide. It has a well-characterized surface (S)-layer, a para-crystalline proteinaceous layer surrounding the cell wall. In many bacterial and archaeal species, the S-layer is glycosylated, but no such modifications have been demonstrated in C. difficile. Here, we show that a C. difficile strain of S-layer cassette type 11, Ox247, has a complex glycan attached via an O-linkage to Thr-38 of the S-layer low-molecular-weight subunit. Using MS and NMR, we fully characterized this glycan. We present evidence that it is composed of three domains: (i) a core peptide–linked tetrasaccharide with the sequence -4-α-Rha-3-α-Rha-3-α-Rha-3-β-Gal-peptide; (ii) a repeating pentasaccharide with the sequence -4-β-Rha-4-α-Glc-3-β-Rha-4-(α-Rib-3-)β-Rha-; and (iii) a nonreducing end–terminal 2,3 cyclophosphoryl-rhamnose attached to a ribose-branched sub-terminal rhamnose residue. The Ox247 genome contains a 24-kb locus containing genes for synthesis and protein attachment of this glycan. Mutations in genes within this locus altered or completely abrogated formation of this glycan, and their phenotypes suggested that this S-layer modification may affect sporulation, cell length, and biofilm formation of C. difficile. In summary, our findings indicate that the S-layer protein of SLCT-11 strains displays a complex glycan and suggest that this glycan is required for C. difficile sporulation and control of cell shape, a discovery with implications for the development of antimicrobials targeting the S-layer.
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Nelsen, Naoma S., Zhigang Li, April L. Warner, Benjamin F. Matthews, and Halina T. Knap. "Genomic Polymorphism Identifies a Subtilisin‐Like Protease near the Rhg4 Locus in Soybean." Crop Science 44, no. 1 (January 2004): 265–73. http://dx.doi.org/10.2135/cropsci2004.2650.
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Richardson, Jason S., Michael F. Hynes, and Ivan J. Oresnik. "A Genetic Locus Necessary for Rhamnose Uptake and Catabolism in Rhizobium leguminosarum bv. trifolii." Journal of Bacteriology 186, no. 24 (December 15, 2004): 8433–42. http://dx.doi.org/10.1128/jb.186.24.8433-8442.2004.
Повний текст джерелаАнотація:
ABSTRACT Rhizobium leguminosarum bv. trifolii mutants unable to catabolize the methyl-pentose rhamnose are unable to compete effectively for nodule occupancy. In this work we show that the locus responsible for the transport and catabolism of rhamnose spans 10,959 bp. Mutations in this region were generated by transposon mutagenesis, and representative mutants were characterized. The locus contains genes coding for an ABC-type transporter, a putative dehydrogenase, a probable isomerase, and a sugar kinase necessary for the transport and subsequent catabolism of rhamnose. The regulation of these genes, which are inducible by rhamnose, is carried out in part by a DeoR-type negative regulator (RhaR) that is encoded within the same transcript as the ABC-type transporter but is separated from the structural genes encoding the transporter by a terminator-like sequence. RNA dot blot analysis demonstrated that this terminator-like sequence is correlated with transcript attenuation only under noninducing conditions. Transport assays utilizing tritiated rhamnose demonstrated that uptake of rhamnose was inducible and dependent upon the presence of the ABC transporter at this locus. Phenotypic analyses of representative mutants from this locus provide genetic evidence that the catabolism of rhamnose differs from previously described methyl-pentose catabolic pathways.
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Liu, Xiaohong, Shiming Liu, Aziz Jamai, Abdelhafid Bendahmane, David A. Lightfoot, Melissa G. Mitchum, and Khalid Meksem. "Soybean cyst nematode resistance in soybean is independent of the Rhg4 locus LRR-RLK gene." Functional & Integrative Genomics 11, no. 4 (May 4, 2011): 539–49. http://dx.doi.org/10.1007/s10142-011-0225-4.
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10
Lewers, K. S., S. D. Nilmalgoda, A. L. Warner, H. T. Knap, and B. F. Matthews. "Physical mapping of resistant and susceptible soybean genomes near the soybean cyst nematode resistance gene Rhg4." Genome 44, no. 6 (December 1, 2001): 1057–64. http://dx.doi.org/10.1139/g01-109.
Повний текст джерелаАнотація:
The soybean cyst nematode (SCN), Heterodera glycines Ichinohe, is the foremost pest of soybean (Glycine max L. Merr.). The rhg1 allele on linkage group (LG) G and the Rhg4 allele on LG A2 are important in conditioning resistance. Markers closely linked to the Rhg4 locus were used previously to screen a library of bacterial artificial chromosome (BAC) clones from susceptible 'Williams 82' and identified a single 150-kb BAC, Gm_ISb001_056_G02 (56G2). End-sequenced subclones positioned onto a restriction map provided landmarks for identifying the corresponding region from a BAC library from accession PI 437654 with broad resistance to SCN. Seventy-three PI 437654 BACs were assigned to contigs based upon HindIII restriction fragment profiles. Four contigs represented the PI 437654 counterpart of the 'Williams 82' BAC, with PCR assays connecting these contigs. Some of the markers on the PI 437654 contigs are separated by a greater physical distance than in the 'Williams 82' BAC and some primers amplify bands from BACs in the mid-portion of the connected PI 437654 BAC contigs that are not amplified from the 'Williams 82' BAC. These observations suggest that there is an insertion in the PI 437654 genome relative to the 'Williams 82' genome in the Rhg4 region.Key words: BAC, deletion, insertion, resistance gene, soybean cyst nematode.
11
Ramazanova, S. A., E. V. Badyanov, V. G. Savichenko, S. Z. Guchetl, and E. A. Strelnikov. "Assessment of linkage between the PLarg, gene controlling resistance to downy mildew in sunflower and microsatellite DNA loci." Oil Crops 3, no. 191 (November 25, 2022): 14–23. http://dx.doi.org/10.25230/2412-608x-2022-3-191-14-23.
Повний текст джерелаАнотація:
Downy mildew caused by oomycete Plasmopara halstedii (Farl.) Berl. et de Toni is one of the most harmful diseases of sunflower. The Plarg locus, which provides resistance to all known races of P. halstedii, is currently promising for use in breeding. This gene is introgressed from the wild species Helianthus argo-phyllus. Microsatellite markers (SSRs) make it possible to control the transfer of genes controlling resistance in breeding material. However, validation of the marker is needed to prove its reliability in identifying genes. We selected the SSR markers ORS 662, ORS 509 and ORS 822 to identify the Plarg gene. We conducted their validation on hybrid combinations of sunflower lines of the breeding of V.S. Pustovoit All-Russian Research Institute of Oil Crops VK 776 and VK 925 with sunflower line RHA 419, which is a do-nor of Plarg gene. It was preliminarily established that these lines differ from each other by the allelic state of these loci. A study of the F1 generation showed that all three microsatellite loci are inherited codominantly. The F2 generation obtained by self-pollination was analyzed by phytopathological methods for resistance to P. halstedii. Phenotype splitting analysis of both crossing combinations showed that the actually observed splitting corresponded to the theoretically expected 3 : 1 pattern in monogenic inheritance of the trait. Analysis of linkage of gene and microsatellite loci showed that the recombination frequencies in the RHA 419 × VK 776 cross was 0.11 for the ORS 662 locus, 0.23 for ORS 509 and 0.31 for ORS 822. And for the combination of RHA 419 × VK 925 cross this indicator was: for ORS 509 – 0.28, for ORS 662 – 0.34. Based on the obtained data, we concluded that three studied microsatellite loci should be used in marker-assisted selection (MAS) of sunflower for resistance to downy mildew pathogen.
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Alarcón, Antonio, and Finnur Lárusson. "Representing de Rham cohomology classes on an open Riemann surface by holomorphic forms." International Journal of Mathematics 28, no. 09 (August 2017): 1740004. http://dx.doi.org/10.1142/s0129167x17400043.
Повний текст джерелаАнотація:
Let [Formula: see text] be a connected open Riemann surface. Let [Formula: see text] be an Oka domain in the smooth locus of an analytic subvariety of [Formula: see text], [Formula: see text], such that the convex hull of [Formula: see text] is all of [Formula: see text]. Let [Formula: see text] be the space of nondegenerate holomorphic maps [Formula: see text]. Take a holomorphic 1-form [Formula: see text] on [Formula: see text], not identically zero, and let [Formula: see text] send a map [Formula: see text] to the cohomology class of [Formula: see text]. Our main theorem states that [Formula: see text] is a Serre fibration. This result subsumes the 1971 theorem of Kusunoki and Sainouchi that both the periods and the divisor of a holomorphic form on [Formula: see text] can be prescribed arbitrarily. It also subsumes two parametric h-principles in minimal surface theory proved by Forstnerič and Lárusson in 2016.
13
Ghassemi, Farshid, and Peter M. Gresshoff. "The Early enod2 and the Leghemoglobin (lbc3) Genes Segregate Independently from Other Known Soybean Symbiotic Genes." Molecular Plant-Microbe Interactions® 11, no. 1 (January 1998): 6–13. http://dx.doi.org/10.1094/mpmi.1998.11.1.6.
Повний текст джерелаАнотація:
Recombinant inbred lines (RILs) as well as an F2 segregating population of soybean Glycine max facilitated the mapping of two expressed sequence tags involved in early nodulation and subsequent nitrogen fixation in soybean. For the early nodulin gene enod2, the parents of RILs, Minsoy and Noir1, showed a polymorphism (5.5 vs 5.9 kb) after EcoRV digestion. Restriction fragment length polymorphism (RFLP) patterns of 42 RILs were analyzed with the MAPMAKER program, linking enod2 to the seed coat color gene, I, with a distance of 11.1 cM on linkage group U3 of RIL map. enod2 and I are located close to Rhg4, a soybean cyst nematode resistance gene, and a locus for seed coat hardness. The molecular marker pA-110 and seed coat color were used to integrate enod2 on an F2 segregating population (72 plants) generated from a cross between cv. Bragg and G. soja PI468.397. enod2 was mapped in the same order as on the RIL map but 18.5 cM from the I locus on the TN map. A microsatellite from the 5′ region of enod2b was mapped in the same position, demonstrating that enod2b and not enod2a was mapped. An RFLP for lbc3 (leghemoglobin) segregated independently from enod2 and the nts-1 supernodulating locus suggesting that in soybean symbiotically significant loci (including rj1, Rj2, and rj6) are not clustered.
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Kasimova, Anastasiya A., Nikolay P. Arbatsky, Jacob Tickner, Johanna J. Kenyon, Ruth M. Hall, Michael M. Shneider, Alina A. Dzhaparova, et al. "Acinetobacter baumannii K106 and K112: Two Structurally and Genetically Related 6-Deoxy-l-talose-Containing Capsular Polysaccharides." International Journal of Molecular Sciences 22, no. 11 (May 26, 2021): 5641. http://dx.doi.org/10.3390/ijms22115641.
Повний текст джерелаАнотація:
Whole genome sequences of two Acinetobacter baumannii clinical isolates, 48-1789 and MAR24, revealed that they carry the KL106 and KL112 capsular polysaccharide (CPS) biosynthesis gene clusters, respectively, at the chromosomal K locus. The KL106 and KL112 gene clusters are related to the previously described KL11 and KL83 gene clusters, sharing genes for the synthesis of l-rhamnose (l-Rhap) and 6-deoxy-l-talose (l-6dTalp). CPS material isolated from 48-1789 and MAR24 was studied by sugar analysis and Smith degradation along with one- and two-dimensional 1H and 13C NMR spectroscopy. The structures of K106 and K112 oligosaccharide repeats (K units) l-6dTalp-(1→3)-D-GlcpNAc tetrasaccharide fragment share the responsible genes in the respective gene clusters. The K106 and K83 CPSs also have the same linkage between K units. The KL112 cluster includes an additional glycosyltransferase gene, Gtr183, and the K112 unit includes α l-Rhap side chain that is not found in the K106 structure. K112 further differs in the linkage between K units formed by the Wzy polymerase, and a different wzy gene is found in KL112. However, though both KL106 and KL112 share the atr8 acetyltransferase gene with KL83, only K83 is acetylated.
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Koukleva, L. M., G. A. Eroshenko, V. E. Kouklev, N. Yu Shavina, Ya M. Krasnov, N. P. Guseva, and V. V. Kutyrev. "A study of nucleotide sequence variability of rha locus genes of Yersinia pestis main and nonmain subspecies." Molecular Genetics, Microbiology and Virology 23, no. 2 (June 2008): 77–82. http://dx.doi.org/10.3103/s0891416808020043.
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Wu, Ding-Tao, Kang-Lin Feng, Ling Huang, Ren-You Gan, Yi-Chen Hu, and Liang Zou. "Deep Eutectic Solvent-Assisted Extraction, Partially Structural Characterization, and Bioactivities of Acidic Polysaccharides from Lotus Leaves." Foods 10, no. 10 (September 30, 2021): 2330. http://dx.doi.org/10.3390/foods10102330.
Повний текст джерелаАнотація:
Lotus leaves are often discarded as byproducts in the lotus industry. Polysaccharides are regarded as one of the essentially bioactive components in lotus leaves. Therefore, in order to promote the application of lotus leaves in the functional food industry, the deep eutectic solvent (DES) assisted extraction of polysaccharides from lotus leaves (LLPs) was optimized, and structural and biological properties of LLPs extracted by DES and hot water were further investigated. At the optimal extraction conditions (water content of 61.0% in DES, extraction temperature of 92 °C, liquid-solid ratio of 31.0 mL/g and extraction time of 126 min), the maximum extraction yield (5.38%) was obtained. Furthermore, LLP-D extracted by DES and LLP-W extracted by hot water possessed the same sugar residues, such as 1,4-α-D-GalAp, 1,4-α-D-GalAMep, 1,3,6-β-D-Galp, 1,4-β-D-Galp, 1,5-α-L-Araf, and 1,2-α-L-Rhap, suggesting the presence of homogalacturonan (HG), rhamnogalacturonan I and arabinogalactan in both LLP-W and LLP-D. Notably, LLP-D was much richer in HG fraction than that of LLP-W, suggesting that the DES could assist to specifically extract HG from lotus leaves. Additionally, the lower molecular weight and higher content of uronic acids were observed in LLP-D, which might contribute to its much stronger in vitro antioxidant, hypoglycemic, and immunomodulatory effects. These findings suggest that the optimized DES assisted extraction method can be a potential approach for specific extraction of acidic polysaccharides with good bioactivities from lotus leaves for applications in the functional food industry.
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Rivers, Damien M. R., Derek D. Kim, and Ivan J. Oresnik. "Inability to Catabolize Rhamnose by Sinorhizobium meliloti Rm1021 Affects Competition for Nodule Occupancy." Microorganisms 10, no. 4 (March 29, 2022): 732. http://dx.doi.org/10.3390/microorganisms10040732.
Повний текст джерелаАнотація:
Rhizobium leguminosarum strains unable to grow on rhamnose as a sole carbon source are less competitive for nodule occupancy. To determine if the ability to use rhamnose as a sole carbon source affects competition for nodule occupancy in Sinorhizobium meliloti, Tn5 mutants unable to use rhamnose as a sole carbon source were isolated. S. meliloti mutations affecting rhamnose utilization were found in two operons syntenous to those of R. leguminosarum. Although the S. meliloti Tn5 mutants were complemented using an R. leguminosarum cosmid that contains the entire wild-type rhamnose catabolic locus, complementation did not occur if the cosmids carried Tn5 insertions within the locus. Through a series of heterologous complementation experiments, enzyme assays, gene fusion, and transport experiments, we show that the S. meliloti regulator, RhaR, is dominant to its R. leguminosarum counterpart. In addition, the data support the hypothesis that the R. leguminosarum kinase is capable of directly phosphorylating rhamnose and rhamnulose, whereas the S. meliloti kinase does not possess rhamnose kinase activity. In nodule competition assays, S. meliloti mutants incapable of rhamnose transport were shown to be less competitive than the wild-type and had a decreased ability to bind plant roots in the presence of rhamnose. The data suggests that rhamnose catabolism is a general determinant in competition for nodule occupancy that spans across rhizobial species.
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Carulli, J. P., and D. L. Hartl. "Variable rates of evolution among Drosophila opsin genes." Genetics 132, no. 1 (September 1, 1992): 193–204. http://dx.doi.org/10.1093/genetics/132.1.193.
Повний текст джерелаАнотація:
Abstract DNA sequences and chromosomal locations of four Drosophila pseudoobscura opsin genes were compared with those from Drosophila melanogaster, to determine factors that influence the evolution of multigene families. Although the opsin proteins perform the same primary functions, the comparisons reveal a wide range of evolutionary rates. Amino acid identities for the opsins range from 90% for Rh2 to more than 95% for Rh1 and Rh4. Variation in the rate of synonymous site substitution is especially striking: the major opsin, encoded by the Rh1 locus, differs at only 26.1% of synonymous sites between D. pseudoobscura and D. melanogaster, while the other opsin loci differ by as much as 39.2% at synonymous sites. Rh3 and Rh4 have similar levels of synonymous nucleotide substitution but significantly different amounts of amino acid replacement. This decoupling of nucleotide substitution and amino acid replacement suggests that different selective pressures are acting on these similar genes. There is significant heterogeneity in base composition and codon usage bias among the opsin genes in both species, but there are no consistent relationships between these factors and the rate of evolution of the opsins. In addition to exhibiting variation in evolutionary rates, the opsin loci in these species reveal rearrangements of chromosome elements.
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Ramazanova, S. A., E. V. Badyanov, S. V. Ivanov, V. G. Savichenko, and S. Z. Guchetl. "Linked inheritance of the microsatellite locus ORS 822 with the gene Plarg controlling the resistance to downy mildew in sunflower." Oil Crops 4, no. 192 (December 25, 2022): 20–28. http://dx.doi.org/10.25230/2412-608x-2022-4-192-20-28.
Повний текст джерелаАнотація:
Resistance to the downy mildew pathogen Plasmopara halstedii (Farl.) Berlese et de Toni in sunflower is controlled by the resistance gene Pl. The Plarg gene is currently promising in breeding for resistance as it is effective against all known races of the pathogen. This gene is introgressed from the wild species Helianthus argophyllus. Molecular markers and, in particular, simple microsatellite repeats (SSRs) allow to control the transfer and pyramiding of disease resistance genes. However, validation of the molecular marker is needed to prove its reliability. The microsatellite marker ORS 822 was tested to identify the Plarg gene. We conducted the research on a hybrid combination of a susceptible sunflower line of VNIIMK's breeding VK 925 and a resistant line RHA 419, a donor of the Plarg gene. We es-tablished that these lines differ from each other by the allelic state of this locus. Molecular analysis of the F1 generation showed that the microsatellite locus is inherited codominantly. We obtained the F2 generation by self-pollination and made a phytopathological evaluation of resistance to P. halstedii. Split analysis by ph-notype showed that the actually observed segregation corresponded to the theoretically expected 3:1 model for monogenic inheritance of the trait. Based on the obtained data, we determined that the Plarg gene and microsatellite locus ORS 822 are linked with a recombination frequency of 0.26. As a result of this research, we concluded that this marker can be used to select homozygous resistant sunflower plants for resistance to downy mildew in marker-assisted selection (MAS).
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Van Calsteren, Marie-Rose, Fleur Gagnon, Sonia Lacouture, Nahuel Fittipaldi, and Marcello Gottschalk. "Structure determination ofStreptococcus suisserotype 2 capsular polysaccharide." Biochemistry and Cell Biology 88, no. 3 (June 2010): 513–25. http://dx.doi.org/10.1139/o09-170.
Повний текст джерелаАнотація:
The capsular polysaccharide (CPS) of Streptococcus suis serotype 2 was isolated, purified, chemically modified, and characterized. Sugar and absolute configuration analyses of the CPS gave the following composition: d-Gal, 3; d-Glc, 1; d-GlcNAc, 1; d-Neu5Ac, 1; l-Rha, 1. Sialic acid was found to be terminal, and the CPS was quantitatively desialylated by mild acid hydrolysis. The CPS was also submitted to periodate oxidation followed by borohydride reduction and Smith degradation. Sugar and methylation analysis,1H and13C nuclear magnetic resonance, and mass spectrometry of the native CPS or of its specifically modified products allowed to determine the repeating unit sequence: [4)[Neu5Ac(α2–6)Gal(β1–4)GlcNAc(β1–3)]Gal(β1–4)[Gal(α1–3)]Rha(β1–4)Glc(β1-]n. The backbone sequence was found to be identical to that of Streptococcus agalactiae or group B Streptococcus (GBS) type VIII and Streptococcus pneumoniae type 23F. The S. suis CPS shares the sequence Neu5Ac-Gal-GlcNAc-Gal in common with GBS types Ia, Ib, II, III, and IV CPSs but differs from them by the presence of rhamnose and the fact that sialic acid is 2,6- rather than 2,3-linked to the following Gal. A correlation between the S. suis CPS sequence and genes of the serotype 2 cps locus encoding putative enzymes responsible for the biosynthesis of the repeating unit was tentatively established.
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Reid, Marion E., Karina A. Rosa, Vivien I. Powell, Christine Lomas-Francis, Fernando F. Costa, Servio T. Stinghen, Alexandra M. Watanabe, et al. "Rhnull Syndrome: Identification of a Novel Mutation in RHce Gene." Blood 104, no. 11 (November 16, 2004): 2708. http://dx.doi.org/10.1182/blood.v104.11.2708.2708.
Повний текст джерелаАнотація:
Abstract Background: Rhnull syndrome, which includes the amorph and regulator types, is a rare genetic disorder characterized by stomatocytosis and chronic mild hemolytic anemia. The suppression of Rh antigen expression for regulator types is attributed to mutations of the RHAG gene. The deficiency of Rh proteins on the red blood cells (RBCs) from the rare individuals of the Rhnull amorph type may be the result of homozygosity for a silent RHCE allele at the RH locus in which the RHD is absent. We studied a Brazilian family transmitting an amorph Rhnull disease gene on a consanguineous background and identified a novel mutation in RHce gene causing the loss of function phenotype. Methods: RBCs from two Rhnull sisters (G1 and G2) of the amorph type and from family members were analyzed by serology and flow cytometry with monoclonal antibodies specific for Rh (D, C, c, E and e), and LW antigens, for RhAG and CD47 as well as alloantibodies directed against GPB (S, s and U) antigens. Genomic DNA samples and transcripts were tested by PCR and sequence analysis. Results: RBCs from G1 and G2 did not react with the anti-Rh and anti-LW reagents and reacted weakly with anti-RhAG, anti-CD47, anti-s, and anti-U reagents. RBCs from G1 were S+ and from G2 S-. Molecular analyses showed a deletion of the nucleotide at position 963 in exon 7 of the RHce gene [GGG(Glu321→GG)] in addition to a deletion of the RHD gene. This nucleotide deletion in exon 7 introduced a frameshift after Glu321 and a premature stop codon, resulting in a shorter predicted protein with 359 amino acids, including a new C-terminal sequence that is likely to alter the protein conformation and impair the Rh complex assembly. Conclusion: We describe a novel mutation of an amorph Rhnull disease gene leading to a loss of function phenotype. Our findings reinforce previous studies suggesting that protein folding and/or protein-protein interaction mediated by the C-terminal domain of the Rh proteins plays a role in the stability of the Rh membrane complex.
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Matthews, B. F., M. H. MacDonald, J. S. Gebhardt, and T. E. Devine. "Molecular markers residing close to the Rhg4 locus conferring resistance to soybean cyst nematode race 3 on linkage group A of soybean." Theoretical and Applied Genetics 97, no. 7 (November 1998): 1047–52. http://dx.doi.org/10.1007/s001220050990.
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Arango Pinedo, Catalina, and Daniel J. Gage. "Plasmids That Insert into the Rhamnose Utilization Locus, rha: A Versatile Tool for Genetic Studies in Sinorhizobium meliloti." Journal of Molecular Microbiology and Biotechnology 17, no. 4 (2009): 201–10. http://dx.doi.org/10.1159/000242446.
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Lakhssassi, Naoufal, Dounya Knizia, Abdelhalim El Baze, Aicha Lakhssassi, Jonas Meksem, and Khalid Meksem. "Proteomic, Transcriptomic, Mutational, and Functional Assays Reveal the Involvement of Both THF and PLP Sites at the GmSHMT08 in Resistance to Soybean Cyst Nematode." International Journal of Molecular Sciences 23, no. 19 (September 24, 2022): 11278. http://dx.doi.org/10.3390/ijms231911278.
Повний текст джерелаАнотація:
The serine hydroxymethyltransferase (SHMT; E.C. 2.1.2.1) is involved in the interconversion of serine/glycine and tetrahydrofolate (THF)/5,10-methylene THF, playing a key role in one-carbon metabolism, the de novo purine pathway, cellular methylation reactions, redox homeostasis maintenance, and methionine and thymidylate synthesis. GmSHMT08 is the soybean gene underlying soybean cyst nematode (SCN) resistance at the Rhg4 locus. GmSHMT08 protein contains four tetrahydrofolate (THF) cofactor binding sites (L129, L135, F284, N374) and six pyridoxal phosphate (PLP) cofactor binding/catalysis sites (Y59, G106, G107, H134, S190A, H218). In the current study, proteomic analysis of a data set of protein complex immunoprecipitated using GmSHMT08 antibodies under SCN infected soybean roots reveals the presence of enriched pathways that mainly use glycine/serine as a substrate (glyoxylate cycle, redox homeostasis, glycolysis, and heme biosynthesis). Root and leaf transcriptomic analysis of differentially expressed genes under SCN infection supported the proteomic data, pointing directly to the involvement of the interconversion reaction carried out by the serine hydroxymethyltransferase enzyme. Direct site mutagenesis revealed that all mutated THF and PLP sites at the GmSHMT08 resulted in increased SCN resistance. We have shown the involvement of PLP sites in SCN resistance. Specially, the effect of the two Y59 and S190 PLP sites was more drastic than the tested THF sites. This unprecedented finding will help us to identify the biological outcomes of THF and PLP residues at the GmSHMT08 and to understand SCN resistance mechanisms.
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Suzuki, Chika, Fumio Taguchi-Shiobara, Chiaki Ikeda, Masao Iwahashi, Takumi Matsui, Yoko Yamashita, and Reina Ogura. "Mapping soybean rhg2 locus, which confers resistance to soybean cyst nematode race 1 in combination with rhg1 and Rhg4 derived from PI 84751." Breeding Science 70, no. 4 (2020): 474–80. http://dx.doi.org/10.1270/jsbbs.20035.
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Perota, A., I. Lagutina, R. Duchi, P. Turini, G. Crotti, S. Colleoni, G. Lazzari, F. Lucchini, and C. Galli. "215 LIVE PIGLETS GENERATED BY SOMATIC CELL NUCLEAR TRANSFER FOLLOWING TARGETING OF A PORCINE ENHANCED GREEN FLUORESCENT PROTEIN LINE MEDIATED BY ZINC-FINGER NUCLEASES TO ESTABLISH CLONED HYGROMYCIN-RESISTANT PRIMARY CELL LINES SUITABLE FOR Cre-MEDIATED RECOMBINASE-MEDIATED CASSETTE EXCHANGE." Reproduction, Fertility and Development 26, no. 1 (2014): 221. http://dx.doi.org/10.1071/rdv26n1ab215.
Повний текст джерелаАнотація:
Recently, site-specific nucleases (zinc-finger nucleases, ZFN; TAL effector nucleases; and CRISPR) emerged as powerful tools for gene modification of different cells types and enhanced green fluorescent protein (EGFP)-specific ZFN were successfully used in the rat (Geurtz et al. 2010) and in the pig (Watanabe et al. 2010; Whyte et al. 2010). Previously (Brunetti et al. 2008 Clon. Stem Cells), we generated an EGFP transgenic porcine line (Verro2GFP) characterised by a single integration of pCAGGS-EGFP cassette, high ubiquitous EGFP expression, Mendelian transgene transmission, and expression in F1. The aim of this work was to modify a transcriptionally active GFP-locus into one suitable for Cre-mediated recombinase-mediated cassette exchange (RMCE), using EGFP-specific ZFN. Homology arms for promoter-less targeting vector were derived from pCAGGS-EGFP vector (promoter fragment = left-homology-arm = LHA; polyA sequence = right-homology-arm = RHA). Cloning floxed (lox2272/lox5171) hygromycin resistance coding sequence between LHA and RHA sequences, we generated the targeting/RMCE vector (pB5′3′Hygro-PL) and its positive control (C+) for PCR set-up (100–1000 plasmid copies). Verro2GFP fibroblasts cultured in DMEM+M199(1 : 1) + 10% FCS, bFGF in 5% CO2, 5% O2, were transfected using Nucleofector (V-024 program). In ZFN-mediated gene targeting, 2 μg of each ZFN coding vector (Sigma-CompoZr®) and 2 μg of pB5′3′Hygro-PL/KpnI vector were used to “nucleofect” 1.4 × 106 Verro2GFP fibroblasts in 2 experiments. Transfected cells were plated in 20 Petri dishes (Ø = 150 mm) and cultured under hygromycin selection (200 μg mL–1) for 15 days. After 12 days of drug selection, 82 resistant colonies were picked up and expanded in 24 multiwell plates for SCNT. All colonies were PCR screened and 45 (54.9%) colonies were positive. Four colonies were used in zona-free SCNT experiments with 140 Day 6 compacted morulae/blastocysts transferred into 2 synchronized sows that both became pregnant. One pregnancy went to term and delivered 5 live animals and 5 stillborn with correct hygromycin cassette integration, detected by PCR. The PCR products were sequenced in 7 animals to verify integration of promoterless targeting vector and in all 7 sequenced samples we obtained a correct insertion without any substitution/deletion. Using hygromycin selection in these experiments, we demonstrated that ZFN-mediated gene targeting can be easily done with high efficiency and is compatible with living animals. Moreover, we have validated a feasible SCNT-tested platform for further Cre-mediated site-specific gene modifications. This work is supported by a grant (Superpig) co-financed by Lombardy Region through the Fund for Promoting Institutional Agreements.
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Zatserklyana, Oleksandra, Khalid Meksem, and David A. Lightfoot. "Additional Polymorphisms Linked to Soybean Cyst Nematode Resistance At The Rhg4 Locus." Atlas Journal of Biology, 2017, 376–83. http://dx.doi.org/10.5147/ajb.v0i0.46.
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Arends, D. W., W. R. Miellet, J. D. Langereis, T. H. A. Ederveen, C. E. van der Gaast - de Jongh, M. van Scherpenzeel, M. J. Knol, et al. "Examining the distribution and impact of single nucleotide polymorphisms in the capsular locus of Streptococcus pneumoniae serotype 19A." Infection and Immunity, July 12, 2021. http://dx.doi.org/10.1128/iai.00246-21.
Повний текст джерелаАнотація:
Streptococcus pneumoniae serotype 19A prevalence has increased after implementation of PCV7 and PCV10 vaccines. In this study, we have provided, with high accuracy, the genetic diversity of the 19A serotype in a cohort of Dutch invasive pneumococcal disease patients and asymptomatic carriers obtained in the period 2004-2016. Whole genomes of the 338 pneumococcal isolates in this cohort were sequenced and their capsule ( cps ) loci compared to examine the diversity and determine the impact on the production of CPS sugar precursors and CPS shedding. We discovered 79 types with a unique CPS locus sequence. Most variation was observed in the rmlB and rmlD genes of the TDP-Rha synthesis pathway, and in the wzg gene, of unknown function. Interestingly, gene variation in the cps locus was conserved in multiple alleles. Using RmlB and RmlD protein models, we predict that enzymatic function is not affected by the single nucleotide polymorphisms as identified. To determine if RmlB and RmlD function was affected, we analyzed nucleotide sugar levels using UHPLC-MS. CPS precursors differed between 19A cps locus subtypes, including TDP-Rha, but no clear correlation was observed. Also, a significant difference in multiple nucleotide sugar levels was observed between phylogenetically branched groups. Because of indications of a role for Wzg in capsule shedding, we analyzed if this was affected. No clear indication of a direct role in shedding was found. We thus describe genotypic variety in rmlB , rmlD and wzg in serotype 19A the Netherlands, for which we have not discovered an associated phenotype.
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Wang, Wenqian, Guangzu Du, Guangyuan Yang, Ke Zhang, Bin Chen та Guanli Xiao. "A multifunctional enzyme portfolio for α-chaconine and α-solanine degradation in the Phthorimaea operculella gut bacterium Glutamicibacter halophytocola S2 encoded in a trisaccharide utilization locus". Frontiers in Microbiology 13 (12 жовтня 2022). http://dx.doi.org/10.3389/fmicb.2022.1023698.
Повний текст джерелаАнотація:
Steroidal glycoalkaloids (SGAs) are secondary metabolites commonly found in members of the family Solanaceae, including potatoes, and are toxic to pests and humans. The predominant SGAs in potato are α-chaconine and α-solanine. We previously reported that Glutamicibacter halophytocola S2, a gut bacterium of the pest Phthorimaea operculella (potato tuber moth), can degrade α-chaconine and α-solanine in potatoes, which can improve the fitness of P. operculella to feed on potatoes with a high content of toxic SGAs. Glutamicibacter halophytocola S2 harbored a gene cluster containing three deglycosylase genes—GE000599, GE000600, and GE000601—that were predicted encode α-rhamnosidase (RhaA), β-glucosidase (GluA), and β-galactosidase (GalA). However, there is limited information is available on the enzyme activities of the three enzymes expressed by this gene cluster and how they degrade the major toxic α-chaconine and α-solanine. In the current study, each enzyme of this gene cluster was produced by a prokaryotic expression approach and the activity of the recombinant enzymes for their target substrate and α-chaconine and α-solanine were evaluated by EPOCH microplate spectrophotometer and liquid chromatography mass spectrometry (LC-MS). The three enzymes had multifunctional activities, with RhaA and GluA could hydrolyze α-rhamnose, β-glucose, and β-galactose, while GalA can hydrolyze β-glucose and β-galactose. The degradation of α-chaconine and α-solanine was consistent with the results of the enzyme activity assays. The final product solanidine could be generated by adding RhaA or GluA alone. In conclusion, this study characterized the multifunctional activity and specific degradation pathway of these three enzymes in G. halophytocola S2. The three multifunctional enzymes have high glycosidic hydrolysis activity and clear gene sequence information, which help facilitates understanding the detoxification mechanism of insect gut microbes. The enzymes have a broad application potential and may be valuable in the removal of toxic SGAs from for potato food consumption.
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Ben Moussa, Hajar, Jacques Pédron, Claire Bertrand, Amandine Hecquet, and Marie-Anne Barny. "Pectobacterium quasiaquaticum sp. nov., isolated from waterways." International Journal of Systematic and Evolutionary Microbiology 71, no. 10 (October 11, 2021). http://dx.doi.org/10.1099/ijsem.0.005042.
Повний текст джерелаАнотація:
Through this study, we established the taxonomic status of seven strains belonging to the genus Pectobacterium (A477-S1-J17T, A398-S21-F17, A535-S3-A17, A411-S4-F17, A113-S21-F16, FL63-S17 and FL60-S17) collected from four different river streams and an artificial lake in south-east France between 2016 and 2017. Ecological surveys in rivers and lakes pointed out different repartition of strains belonging to this clade compared to the closest species, Pectobacterium aquaticum . The main phenotypic difference observed between these strains and the P. aquaticum type strain was strongly impaired growth with rhamnose as the sole carbon source. This correlates with three different forms of pseudogenization of the l-rhamnose/proton symporter gene rhaT in the genomes of strains belonging to this clade. Phylogenetic analysis using gapA gene sequences and multi locus sequence analysis of the core genome showed that these strains formed a distinct clade within the genus Pectobacterium closely related to P. aquaticum. Digital DNA–DNA hybridization (dDDH) and average nucleotide identity (ANI) values showed a clear discontinuity between the new clade and P. aquaticum . However, the calculated values are potentially consistent with either splitting or merging of this new clade with P. aquaticum . In support of the split, ANI coverages were higher within this new clade than between this new clade and P. aquaticum . The split is also consistent with the range of observed ANI or dDDH values that currently separate several accepted species within the genus Pectobacterium . On the basis of these data,strains A477-S1-J17T, A398-S21-F17, A535-S3-A17, A411-S4-F17, A113-S21-F16, FL63-S17 and FL60-S17 represent a novel species of the genus Pectobacterium , for which the name Pectobacterium quasiaquaticum sp. nov. is proposed. The type strain is A477-S1-J17T (=CFBP 8805T=LMG 32181T).
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Sadovskaya, Irina, Evgeny Vinogradov, Pascal Courtin, Julija Armalyte, Mickael Meyrand, Efstathios Giaouris, Simon Palussière, et al. "Another Brick in the Wall: a Rhamnan Polysaccharide Trapped inside Peptidoglycan of Lactococcus lactis." mBio 8, no. 5 (September 12, 2017). http://dx.doi.org/10.1128/mbio.01303-17.
Повний текст джерелаАнотація:
ABSTRACT Polysaccharides are ubiquitous components of the Gram-positive bacterial cell wall. In Lactococcus lactis, a polysaccharide pellicle (PSP) forms a layer at the cell surface. The PSP structure varies among lactococcal strains; in L. lactis MG1363, the PSP is composed of repeating hexasaccharide phosphate units. Here, we report the presence of an additional neutral polysaccharide in L. lactis MG1363 that is a rhamnan composed of α-l-Rha trisaccharide repeating units. This rhamnan is still present in mutants devoid of the PSP, indicating that its synthesis can occur independently of PSP synthesis. High-resolution magic-angle spinning nuclear magnetic resonance (HR-MAS NMR) analysis of whole bacterial cells identified a PSP at the surface of wild-type cells. In contrast, rhamnan was detected only at the surface of PSP-negative mutant cells, indicating that rhamnan is located underneath the surface-exposed PSP and is trapped inside peptidoglycan. The genetic determinants of rhamnan biosynthesis appear to be within the same genetic locus that encodes the PSP biosynthetic machinery, except the gene tagO encoding the initiating glycosyltransferase. We present a model of rhamnan biosynthesis based on an ABC transporter-dependent pathway. Conditional mutants producing reduced amounts of rhamnan exhibit strong morphological defects and impaired division, indicating that rhamnan is essential for normal growth and division. Finally, a mutation leading to reduced expression of lcpA, encoding a protein of the LytR-CpsA-Psr (LCP) family, was shown to severely affect cell wall structure. In lcpA mutant cells, in contrast to wild-type cells, rhamnan was detected by HR-MAS NMR, suggesting that LcpA participates in the attachment of rhamnan to peptidoglycan. IMPORTANCE In the cell wall of Gram-positive bacteria, the peptidoglycan sacculus is considered the major structural component, maintaining cell shape and integrity. It is decorated with other glycopolymers, including polysaccharides, the roles of which are not fully elucidated. In the ovococcus Lactococcus lactis, a polysaccharide with a different structure between strains forms a layer at the bacterial surface and acts as the receptor for various bacteriophages that typically exhibit a narrow host range. The present report describes the identification of a novel polysaccharide in the L. lactis cell wall, a rhamnan that is trapped inside the peptidoglycan and covalently bound to it. We propose a model of rhamnan synthesis based on an ABC transporter-dependent pathway. Rhamnan appears as a conserved component of the lactococcal cell wall playing an essential role in growth and division, thus highlighting the importance of polysaccharides in the cell wall integrity of Gram-positive ovococci. IMPORTANCE In the cell wall of Gram-positive bacteria, the peptidoglycan sacculus is considered the major structural component, maintaining cell shape and integrity. It is decorated with other glycopolymers, including polysaccharides, the roles of which are not fully elucidated. In the ovococcus Lactococcus lactis, a polysaccharide with a different structure between strains forms a layer at the bacterial surface and acts as the receptor for various bacteriophages that typically exhibit a narrow host range. The present report describes the identification of a novel polysaccharide in the L. lactis cell wall, a rhamnan that is trapped inside the peptidoglycan and covalently bound to it. We propose a model of rhamnan synthesis based on an ABC transporter-dependent pathway. Rhamnan appears as a conserved component of the lactococcal cell wall playing an essential role in growth and division, thus highlighting the importance of polysaccharides in the cell wall integrity of Gram-positive ovococci.