Xylan Metabolism Research Articles

CRISPR/Cas9-Mediated Targeted Mutagenesis of GmEOD1 Enhances Seed Size of Soybean

Seed size is a critical agronomic trait that influences the yield and appearance quality of soybeans, making it a primary breeding objective with significant economic value. While the molecular mechanisms that regulate soybean seed size remain largely unknown, several functional molecular targets have been applied in breeding to create larger grain size materials. In this study, we utilized the CRISPR/Cas9 system to induce the targeted mutagenesis of GmEOD1, which encodes the E3 ubiquitin ligase. The resulting homozygous soybean mutant of GmEOD1 exhibited larger seed size and 100-seed weight, with no significant change in the average seed weight per plant. The sum of crude protein and oil content increased significantly in mutants while fatty acid composition remained unchanged. We identified six haplotypes among 156 soybean cultivars, with Hap1 and Hap2 representing the majority of cultivars with relatively higher 100-seed weight, suggesting that sequence variations of GmEOD1 may correlate with seed weight. Transcriptomic analysis across five stages of seed development revealed that stages one–three mainly focused on cell cycle, growth, wall synthesis and modification, photosynthesis, and sugar metabolism; promoting cell growth, reproduction, and carbon accumulation; and providing key intermediates for substance synthesis. Stages four–five focused on polysaccharide catabolism, xylan metabolism, and nutrient pool activity, promoting the accumulation of dry matter, such as sugars, proteins, and lipids in seeds. Weighted gene co-expression network analysis (WGCNA) of modules related to seed size revealed 13 hub genes involved in seed development regulation. This study provides a valuable theoretical basis and excellent opportunities for genetic editing of germplasm cells with subsequent molecular soybean seed size breeding, facilitating easier seed selection to improve soybean quality.

Transcriptome Profiling of Stem-Differentiating Xylem in Response to Abiotic Stresses Based on Hybrid Sequencing in Cunninghamia lanceolata.

Cunninghamia lanceolata (C. lanceolata) belongs to Gymnospermae, which are fast-growing and have desirable wood properties. However, C. lanceolata's stress resistance is little understood. To unravel the physiological and molecular regulation mechanisms under environmental stresses in the typical gymnosperm species of C. lanceolata, three-year-old plants were exposed to simulated drought stress (polyethylene glycol 8000), salicylic acid, and cold treatment at 4 °C for 8 h, 32 h, and 56 h, respectively. Regarding the physiological traits, we observed a decreased protein content and increased peroxidase upon salicylic acid and polyethylene glycol treatment. Superoxide dismutase activity either decreased or increased at first and then returned to normal under the stresses. Regarding the molecular regulation, we used both nanopore direct RNA sequencing and short-read sequencing to reveal a total of 5646 differentially expressed genes in response to different stresses, of which most had functions in lignin catabolism, pectin catabolism, and xylan metabolism, indicating that the development of stem-differentiating xylem was affected upon stress treatment. Finally, we identified a total of 51 AP2/ERF, 29 NAC, and 37 WRKY transcript factors in C. lanceolata. The expression of most of the NAC TFs increased under cold stress, and the expression of most of the WRKY TFs increased under cold and SA stress. These results revealed the transcriptomics responses in C. lanceolata to short-term stresses under this study's experimental conditions and provide preliminary clues about stem-differentiating xylem changes associated with different stresses.

Manipulation of Mini-Yeast Artificial Chromosome Containing Xylan Metabolism Related Genes and Mitotic Stability Analysis in Yeast

Manipulation of Mini-Yeast Artificial Chromosome Containing Xylan Metabolism Related Genes and Mitotic Stability Analysis in Yeast

Thinopyrum intermedium TiAP1 interacts with a chitin deacetylase from Blumeria graminis f. sp. tritici and increases the resistance to Bgt in wheat.

SummaryThe biotrophic fungal pathogen Blumeria graminis f. sp. tritici (Bgt) is a crucial factor causing reduction in global wheat production. Wild wheat relatives, for example Thinopyrum intermedium, is one of the wild‐used parents in wheat disease‐resistant breeding. From T. intermedium line, we identified the aspartic protease gene, TiAP1, which is involved in resistance against Bgt. TiAP1 is a secreted protein that accumulates in large amounts at the infection sites of Bgt and extends to the intercellular space. Yeast two‐hybrid, luciferase complementation imaging and bimolecular florescent complimentary analysis showed that TiAP1 interacted with the chitin deacetylase (BgtCDA1) of Bgt. The yeast expression, purification and in vitro test confirmed the chitin deacetylase activity of BgtCDA1. The bombardment and VIGS‐mediated host‐induced gene silencing showed that BgtCDA1 promotes the invasion of Bgt. Transcriptome analysis showed the cell wall xylan metabolism, lignin biosynthesis‐related and defence genes involved in the signal transduction were up‐regulated in the transgenic TiAP1 wheat induced by Bgt. The TiAP1 in wheat may inactivate the deacetylation function of BgtCDA1, cause chitin oligomers expose to wheat chitin receptor, then trigger the wheat immune response to inhibit the growth and penetration of Bgt, and thereby enhance the resistance of wheat to pathogens.

Whole-Genome Sequence Data Analysis of Anoxybacillus kamchatkensis NASTPD13 Isolated from Hot Spring of Myagdi, Nepal.

Anoxybacillus kamchatkensis NASTPD13 isolated from Paudwar hot spring of Myagdi, Nepal, upon morphological and biochemical analysis revealed to be Gram-positive, straight or slightly curved, rod-shaped, spore-forming, catalase, and oxidase-positive facultative anaerobes. It grows over a wide range of pH (5.0-11) and temperature (37-75°C), which showed growth in different reduced carbon sources such as starch raffinose, glucose, fructose, inositol, trehalose, sorbitol, mellobiose, and mannitol in aerobic conditions. Furthermore, the partial sequence obtained upon sequencing showed 99% sequence similarity in 16S rRNA gene sequence with A. kamchatkensis JW/VK-KG4 and was suggested to be Anoxybacillus kamchatkensis. Moreover, whole-genome analysis of NASTPD13 revealed 2,866,796 bp genome with a G+C content of 41.6%. Analysis of the genome revealed the presence of 102 RNA genes, which includes sequences coding for 19 rRNA and 79 tRNA genes. While the 16S rRNA gene sequence of strain NASTPD13 showed high similarity (>99%) to those of A. kamchatkensis JW/VK-KG4, RAST analysis of NASTPD13 genome suggested that A. kamchatkensis G10 is actually the closest neighbor in terms of sequence similarity. The genome annotation by RAST revealed various genes encoding glycoside hydrolases supporting that it can utilize several reduced carbon sources as observed and these genes could be important for carbohydrate-related industries. Xylanase pathway, particularly the genomic region encoding key enzymes for xylan depolymerization and xylose metabolism, further confirmed the presence of the complete gene in xylan metabolism. In addition, the complete xylose utilization gene locus analysis of NASTPD13 genome revealed all including D-xylose transport ATP-binding protein XylG and XylF, the xylose isomerase encoding gene XylA, and the gene XylB coding for a xylulokinase supported the fact that the isolate contains a complete set of genes related to xylan degradation, pentose transport, and metabolism. The results of the present study suggest that the isolated A. kamchatkensis NASTPD13 containing xylanase-producing genes could be useful in lignocellulosic biomass-utilizing industries where pentose polymers could also be utilized along with the hexose polymers.

The Role of Clt1-Regulated Xylan Metabolism in Melanin and Toxin Formation for the Pathogenicity of Curvularia lunata in Maize.

We previously reported that the BTB (brica-brac, tramtrack, and broad) domain-containing protein Clt1 regulates melanin and toxin synthesis, conidiation, and pathogenicity in Curvularia lunata, but the interacting proteins and regulative mechanism of Clt1 are unclear. In this research, we identified two proteins, which respectively correspond to xylanase (Clxyn24) and acetyl xylan esterase (Claxe43) from C. lunata, that were regulated by Clt1. Yeast two-hybrid (Y2H) and bimolecular fluorescence complementation assays were conducted to verify the interaction of Clt1 with full-length Clxyn24 and Claxe43. Furthermore, the Y2H assay revealed that Clt1 physically interacted with Clxyn24 and Claxe43 through its BTB domain to degrade xylan, which was used as a carbon source for C. lunata growth. The utilization of xylan provides acetyl-CoA for the synthesis of melanin and toxin as well as energy and other intermediate metabolites for conidiation. Furthermore, transcriptome analysis revealed that PKS18 and its 13 flanking genes found clustered in a region spanning 57.89 kb on scaffold 9 of the C. lunata CX-3 genome were down-regulated in toxin production-deficient mutant T806, and this cluster is possibly responsible for toxin biosynthesis of C. lunata.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Chromosome-level draft genome of a diploid plum (Prunus salicina).

BackgroundPlums are one of the most economically important Rosaceae fruit crops and comprise dozens of species distributed across the world. Until now, only limited genomic information has been available for the genetic studies and breeding programs of plums. Prunus salicina, an important diploid plum species, plays a predominant role in modern commercial plum production. Here we selected P. salicina for whole-genome sequencing and present a chromosome-level genome assembly through the combination of Pacific Biosciences sequencing, Illumina sequencing, and Hi-C technology.FindingsThe assembly had a total size of 284.2 Mb, with contig N50 of 1.78 Mb and scaffold N50 of 32.32 Mb. A total of 96.56% of the assembled sequences were anchored onto 8 pseudochromosomes, and 24,448 protein-coding genes were identified. Phylogenetic analysis showed that P. salicina had a close relationship with Prunus mume and Prunus armeniaca, with P. salicina diverging from their common ancestor ∼9.05 million years ago. During P. salicina evolution 146 gene families were expanded, and some cell wall–related GO terms were significantly enriched. It was noteworthy that members of the DUF579 family, a new class involved in xylan biosynthesis, were significantly expanded in P. salicina, which provided new insight into the xylan metabolism in plums.ConclusionsWe constructed the first high-quality chromosome-level plum genome using Pacific Biosciences, Illumina, and Hi-C technologies. This work provides a valuable resource for facilitating plum breeding programs and studying the genetic diversity mechanisms of plums and Prunus species.

Simultaneous and Sequential Integration by Cre/loxP Site-Specific Recombination in Saccharomyces cerevisiae.

A Cre/loxP-δ-integration system was developed to allow sequential and simultaneous integration of a multiple gene expression cassette in Saccharomyces cerevisiae. To allow repeated integrations, the reusable Candida glabrata MARKER (CgMARKER) carrying loxP sequences was used, and the integrated CgMARKER was efficiently removed by inducing Cre recombinase. The XYLP and XYLB genes encoding endoxylanase and β-xylosidase, respectively, were used as model genes for xylan metabolism in this system, and the copy number of these genes was increased to 15.8 and 16.9 copies/cell, respectively, by repeated integration. This integration system is a promising approach for the easy construction of yeast strains with enhanced metabolic pathways through multicopy gene expression.

Low genetic diversity and functional constraint of miRNA genes participating pollen-pistil interaction in rice.

In this study, we sequenced and analyzed the expression and evolution of rice miRNA genes participating pollen-pistil interaction that is crucial to rice yield. Pollen-pistil interaction is an essential reproductive process for all flowering plants. While microRNAs (miRNAs) are important noncoding small RNAs that regulate mRNA levels in eukaryotic cells, there is little knowledge about which miRNAs involved in the early stages of pollen-pistil interaction in rice and how they evolve under this conserved process. In this study, we sequenced the small RNAs in rice from unpollinated pistil (R0), pistil from 5min and 15min after pollination, respectively, to identify known and novel miRNAs that are involved in this process. By comparing the corresponding mRNA-seq dataset, we identified a group of miRNAs with strong negative expression pattern with their target genes. Further investigation of all miRNA loci (MIRNAs) across 1083 public rice accessions revealed significantly reduced genetic diversity in MIRNAs with strong negative expression of their targets when comparing to those with little or no impact on targets during pollen-pistil interaction. Annotation of targets suggested that those MIRNAs with strong impact on targets were pronounced in cell wall related processes such as xylan metabolism. Additionally, plant conserved miRNAs, such as those with functions in gibberellic acid, auxin and nitrate signaling, were also with strong negative expression of their targets. Overall, our analyses identified key miRNAs participating pollen-pistil interaction and their evolutionary patterns in rice, which can facilitate the understanding of molecular mechanisms associated with seed setting.

Network-based integration of systems genetics data reveals pathways associated with lignocellulosic biomass accumulation and processing

As a consequence of their remarkable adaptability, fast growth, and superior wood properties, eucalypt tree plantations have emerged as key renewable feedstocks (over 20 million ha globally) for the production of pulp, paper, bioenergy, and other lignocellulosic products. However, most biomass properties such as growth, wood density, and wood chemistry are complex traits that are hard to improve in long-lived perennials. Systems genetics, a process of harnessing multiple levels of component trait information (e.g., transcript, protein, and metabolite variation) in populations that vary in complex traits, has proven effective for dissecting the genetics and biology of such traits. We have applied a network-based data integration (NBDI) method for a systems-level analysis of genes, processes and pathways underlying biomass and bioenergy-related traits using a segregating Eucalyptus hybrid population. We show that the integrative approach can link biologically meaningful sets of genes to complex traits and at the same time reveal the molecular basis of trait variation. Gene sets identified for related woody biomass traits were found to share regulatory loci, cluster in network neighborhoods, and exhibit enrichment for molecular functions such as xylan metabolism and cell wall development. These findings offer a framework for identifying the molecular underpinnings of complex biomass and bioprocessing-related traits. A more thorough understanding of the molecular basis of plant biomass traits should provide additional opportunities for the establishment of a sustainable bio-based economy.

Expression and characterization of a novel endo-1,4-β-xylanase produced by Streptomyces griseorubens JSD-1 isolated from compost-treated soil

Streptomyces griseorubens JSD-1 is an isolate sourced from compost-treated soil that can utilize xylan as the unique carbon source for growth; xylanlytic genes related to this process were expected to be crucial. However, little is known about the genetic basic of utilizing xylan in this isolate. To further investigate the molecular mechanism of xylan metabolism, genome sequencing was carried out. The draft genome obtained has been deposited with GenBank. Endo-1,4-β-xylanase (Xyn) was acquired, followed by characterization of its cellular localization and expression profiles. Xyn proved to be a novel enzyme, differing from xylanases from similar species according to combined analyses from multiple sequence alignments and putative three-dimensional structure modeling. Moreover, recombinant expression was optimized in heterogeneous hosts to further determine optimum pH and temperature conditions as well as residual activity in the presence of metal ions and inhibitors. Finally, substrate specificity and kinetic parameters were also studied. In conclusion, our findings illustrate the genetic background related to xylan metabolism in this isolate. In addition, a novel Xyn that is expected to have potential in the bio-degradation of hemicellulosic materials was expressed, purified and characterized.

NAD+-dependent xylitol dehydrogenase (xdhA) and l-arabitol-4-dehydrogenase (ladA) deletion mutants of Aspergillus oryzae for improved xylitol production

Xylitol dehydrogenase (XDHA) and L-arabitol dehydrogenase (LADA) are two key enzymes in xylan metabolism catalyzing the oxidation of xylitol to D-xylulose and arabitol to L-xylulose, respectively. In Aspergillus oryzae, XDHA and LADA are encoded by xdhA and ladA. We deleted xdhA and ladA and xdhA-ladA to generate mutants with decreased dehydrogenase activities and increased xylitol production. The mutants were constructed by homologous transformation into A. oryzae P4 (∆pyrG) using pyrG as a selectable marker. The xylitol productivity of the mutants was measured using D-xylose as the sole carbohydrate source. xdhA, ladA, and the double-deletion mutant produced, respectively, 12.4 g xylitol/l with a yield of 0.24 g/g D-xylose, 12.4 g/l with a yield of 0.33 g/g D-xylose, and 8.6 g/l at a yield of 0.26 g/g D-xylose.

Biological conversion assay using Clostridium phytofermentans to estimate plant feedstock quality

BackgroundThere is currently considerable interest in developing renewable sources of energy. One strategy is the biological conversion of plant biomass to liquid transportation fuel. Several technical hurdles impinge upon the economic feasibility of this strategy, including the development of energy crops amenable to facile deconstruction. Reliable assays to characterize feedstock quality are needed to measure the effects of pre-treatment and processing and of the plant and microbial genetic diversity that influence bioconversion efficiency.ResultsWe used the anaerobic bacterium Clostridium phytofermentans to develop a robust assay for biomass digestibility and conversion to biofuels. The assay utilizes the ability of the microbe to convert biomass directly into ethanol with little or no pre-treatment. Plant samples were added to an anaerobic minimal medium and inoculated with C. phytofermentans, incubated for 3 days, after which the culture supernatant was analyzed for ethanol concentration. The assay detected significant differences in the supernatant ethanol from wild-type sorghum compared with brown midrib sorghum mutants previously shown to be highly digestible. Compositional analysis of the biomass before and after inoculation suggested that differences in xylan metabolism were partly responsible for the differences in ethanol yields. Additionally, we characterized the natural genetic variation for conversion efficiency in Brachypodium distachyon and shrub willow (Salix spp.).ConclusionOur results agree with those from previous studies of lignin mutants using enzymatic saccharification-based approaches. However, the use of C. phytofermentans takes into consideration specific organismal interactions, which will be crucial for simultaneous saccharification fermentation or consolidated bioprocessing. The ability to detect such phenotypic variation facilitates the genetic analysis of mechanisms underlying plant feedstock quality.

A Family 8 Glycoside Hydrolase from Bacillus halodurans C-125 (BH2105) Is a Reducing End Xylose-releasing Exo-oligoxylanase

The gene encoding family 8 glycoside hydrolases from Bacillus halodurans C-125 (BH2105), an alkalophilic bacterium with a known genomic sequence, was expressed in Escherichia coli. The protein was expressed with the intact N-terminal sequence, suggesting that it did not possess a signal peptide and that it was an intracellular enzyme. The recombinant enzyme showed no hydrolytic activity on xylan, whereas it had been annotated as xylanase Y. It hydrolyzed xylooligosaccharide whose degree of polymerization is greater than or equal to 3 in an exo-splitting manner with anomeric inversion, releasing the xylose unit at the reducing end. Judging from its substrate specificity and reaction mechanism, we named the enzyme reducing end xylose-releasing exo-oligoxylanase (Rex). Rex was found to utilize only the beta-anomer of the substrate to form beta-xylose and alpha-xylooligosaccharide. The optimum pH of the enzymatic reaction (6.2-7.3) was found in the neutral range, a range beneficial for intracellular enzymes. The genomic sequence suggests that B. halodurans secretes two endoxylanases and possesses two alpha-arabinofuranosidases, one alpha-glucuronidase, and three beta-xylosidases intracellularly in addition to Rex. The extracellular enzymes supposedly hydrolyze xylan into arabino/glucurono-xylooligosaccharides that are then transported into the cells. Rex may play a role as a key enzyme in intracellular xylan metabolism in B. halodurans by cleaving xylooligosaccharides that were produced by the action of other intracellular enzymes from the arabino/glucurono-xylooligosaccharides.

Isolation and characterization of two specific regulatory Aspergillus niger mutants shows antagonistic regulation of arabinan and xylan metabolism.

This paper describes two Aspergillus niger mutants (araA and araB) specifically disturbed in the regulation of the arabinanase system in response to the presence of L-arabinose. Expression of the three known L-arabinose-induced arabinanolytic genes, abfA, abfB and abnA, was substantially decreased or absent in the araA and araB strains compared to the wild-type when incubated in the presence of L-arabinose or L-arabitol. In addition, the intracellular activities of L-arabitol dehydrogenase and L-arabinose reductase, involved in L-arabinose catabolism, were decreased in the araA and araB strains. Finally, the data show that the gene encoding D-xylulose kinase, xkiA, is also under control of the arabinanolytic regulatory system. L-Arabitol, most likely the true inducer of the arabinanolytic and L-arabinose catabolic genes, accumulated to a high intracellular concentration in the araA and araB mutants. This indicates that the decrease of expression of the arabinanolytic genes was not due to lack of inducer accumulation. Therefore, it is proposed that the araA and araB mutations are localized in positive-acting components of the regulatory system involved in the expression of the arabinanase-encoding genes and the genes encoding the L-arabinose catabolic pathway.

Cellulose and Xylan Utilisation in the Lower Termite Reticulitermes speratus

The distribution of the enzymes of cellulose and xylan metabolism namely endo-β-1,4-glucanase, β-glucosidase, endo-β-1,4-xylanase and β-xylosidase activities, in Reticulitermes speratus (Kolbe) was measured both in the salivary glands and in the major gut sections and along the length of the gut in freshly collected termites. The majority of the endo-β-1,4-glucanase activity (77.8%) was found in the salivary glands which also contained 23.9% of the β-glucosidase activity. At least 70% of the remaining activity was located in the anterior section of the hindgut. A small amount of endo-β-1,4-xylanase activity (2.4%), but no β-xylosidase activity, was present in the salivary glands. The majority of these activities were in the anterior section of the hindgut. The RQ of freshly collected termites at 25°C was 1.03±0.01. Maintaining termites for 16 days on wood, cellulose and xylan showed that the RQ values of termites fed on wood or xylan were not significantly different from those of freshly collected termites but significantly increased when maintained on cellulose. The RQ of starved termites after 11 days was 0.81±0.02. There were three effects on protozoan populations of feeding termites xylan for 20 days. One species, Dinenympha parva was not affected, while five others, Pyrsonympha grandis, Holomastigotes elongatum, Dinenympha rugosa, Dinenympha leidy and Dinenympha porteri survived for 20 days but slowly decreased in numbers. The numbers of P. grandis and D. leidy surviving for 20 days were significantly different from those in starved termites. The third group comprising the two large species, Teratonympha mirabilis and Trichonympha agilis and three small species, Pyrsonympha modesta, Dinenympha exilis and Dinenympha nobilis disappeared within 15 days as in starved termites. It is suggested that protozoa in the first two groups are xylanolytic. Protozoan populations on wood and cellulose diets were not markedly affected. Selective removal of the protozoa by u.v. irradiation led to the loss of xylanolytic activity and a life span comparable to starved termites. © 1997 Elsevier Science Ltd. All rights reserved

Effect of heterotrophic ruminal bacteria on xylan metabolism by the anaerobic fungus Piromyces communis

Six rumen bacteria were cocultured with the rumen fungus Piromyces communis and the effects on xylanolysis determined. The rate and extent of xylan utilization was enhanced in cocultures with Prevotella ruminicola or Succinivibrio dextrinosolvens. The positive effects of Suc. dextrinosolvens and Prev. ruminicola on xylanolysis by P. communis correlated with effective cross-feeding by the bacteria on arabinose and xylose released from xylan. Xylanolysis was not enhanced in cocultures of P. communis with Streptococcus bovis, Veillonella parvula or Ruminococcus flavefaciens. A comparison of fermentation product profiles and of extracellular enzyme activities showed that whereas saccharolytic species and Butyrivibrio fibrisolvens were dominant in cocultures, P. communis dominated in the culture with R. flavefaciens. Extracellular xylanase and β-xylosidase activities were not increased by cocultivation of P. communis with any of the heterotrophic bacteria.

Partial purification and properties of an endo‐xylanase from cucumber seeds

An endo‐xylanase. 1,4‐β‐D‐xylan xylanohydrolase (EC 3.2.1.8) from immature cucumber L. cv. Heinz 3534) seeds was partially purified using ammonium sulfate fractionation and chromatography on SP‐Sephadex and Sephadex G‐100 in order to determine its role in xylan metabolism during development. Attempts to further purify the enzyme using chromatography on DEAE‐Sephadex, Bio‐Gel HTP hydroxylapatite, Sephadex G‐200 and con A‐Sepharose 4B and native polyacrylamide gel electrophoresis resulted in a significant decrease or complete loss of enzyme activity. Endo‐xylanase had a native molecular weight of 96 kDa as determined by gel filtration, exhibited optimal activity at pH 5.0 and 48°C, and was most stable from pH 4.0 to 5.0. Using beechwood 4‐o‐methyl‐d‐glucurono‐d‐xylan dyed with Remazol Brilliant Blue R as substrate, the Km was estimated to be 0.70 mg ml−1. HgCl2 at 1 mM inhibited endo‐xylanase completely. Other metal ions inhibited the enzyme in the order Cu2+ > Fe3+ > Zn2+ > Ca2+ > Mn2+. The ethanol‐soluble products of endo‐xylanase action on beechwood xylan were isolated and characterized by consecutive chromatography on Bio‐Gel P‐10 and P‐2. The major reaction products were xylo‐oligosaccharides [degree of polymerization (dp) > 10] but traces of xylobiose and free xylose were also isolated. The formation of xylo‐oligosaccharides indicated that the reaction was catalyzed primarily by an endoxylanase. The partially pure enzyme had no activity towards other cell wall polysaccharides such as cellulose, carboxymethyl cellulose, sodium carboxyl cellulose, potato starch, orange pectin, polygalacturonic acid, arabinogalactan and β‐giucan. However, it was able to hydrolyze larchwood and oat spelts xylan and a polysaccharide component from purified cucumber cell walls. The ability to utilize a substrate from cucumber cell walls supports the hypothesis that endo‐xylanase is involved in the development of cucumber seeds.