Effect of Bran Pre-Treatment with Endoxylanase on the Characteristics of Intermediate Wheatgrass (Thinopyrum intermedium) Bread

Previous work indicated that bran removal promotes network formation in breads prepared from intermediate wheatgrass (IWG) flour. However, refinement reduces yields as well as contents of nutritionally beneficial compounds such as fiber. This study evaluated xylanase pretreatment of IWG bran as a processing option to enhance the properties of bread made with half of the original bran content. Xylanase pretreatment did not affect stickiness but significantly reduced hardness and increased specific loaf volumes compared to negative (without xylanase) and positive controls (with xylanase but without pretreatment). However, the surface of breads with pretreated bran was uneven due to structural collapse during baking. Fewer but larger gas cells were present due to pretreatment. Addition of ascorbic acid modulated these effects, but did not prevent uneven surfaces. Accessible thiol concentrations were slightly but significantly increased by xylanase pretreatment, possibly due to a less compact crumb structure. Endogenous xylanases (apparent activity 0.46 and 5.81 XU/g in flour and bran, respectively) may have been activated during the pretreatment. Moreover, Triticum aestivum xylanase inhibitor activity was also detected (193 and 410 InU/g in flour and bran). Overall, xylanase pretreatment facilitates incorporation of IWG bran into breads, but more research is needed to improve bread appearance.

Genome-Wide Identification of Triticum aestivum Xylanase Inhibitor Gene Family and Inhibitory Effects of XI-2 Subfamily Proteins on Fusarium graminearum GH11 Xylanase

Triticum aestivum xylanase inhibitor (TaXI) gene plays an important role in plant defense. Recently, TaXI–III inhibitor has been shown to play a dual role in wheat resistance to Fusarium graminearum infection. Thus, identifying the members of the TaXI gene family and clarifying its role in wheat resistance to stresses are essential for wheat resistance breeding. However, to date, no comprehensive research on TaXIs in wheat (Triticum aestivum L.) has been conducted. In this study, a total of 277 TaXI genes, including six genes that we cloned, were identified from the recently released wheat genome database (IWGSC RefSeq v1.1), which were unevenly located on 21 chromosomes of wheat. Phylogenetic analysis divided these genes into six subfamilies, all the six genes we cloned belonged to XI-2 subfamily. The exon/intron structure of most TaXI genes and the conserved motifs of proteins in the same subfamily are similar. The TaXI gene family contains 92 homologous gene pairs or clusters, 63 and 193 genes were identified as tandem replication and segmentally duplicated genes, respectively. Analysis of the cis-acting elements in the promoter of TaXI genes showed that they are involved in wheat growth, hormone-mediated signal transduction, and response to biotic and abiotic stresses. RNA-seq data analysis revealed that TaXI genes exhibited expression preference or specificity in different organs and developmental stages, as well as in diverse stress responses, which can be regulated or induced by a variety of plant hormones and stresses. In addition, the qRT-PCR data and heterologous expression analysis of six TaXI genes revealed that the genes of XI-2 subfamily have double inhibitory effect on GH11 xylanase of F. graminearum, suggesting their potential important roles in wheat resistance to F. graminearum infection. The outcomes of this study not only enhance our understanding of the TaXI gene family in wheat, but also help us to screen more candidate genes for further exploring resistance mechanism in wheat.

The Xylanase Inhibitor TAXI-I Increases Plant Resistance to Botrytis cinerea by Inhibiting the BcXyn11a Xylanase Necrotizing Activity

During host plant infection, pathogens produce a wide array of cell wall degrading enzymes (CWDEs) to break the plant cell wall. Among CWDEs, xylanases are key enzymes in the degradation of xylan, the main component of hemicellulose. Targeted deletion experiments support the direct involvement of the xylanase BcXyn11a in the pathogenesis of Botrytis cinerea. Since the Triticum aestivum xylanase inhibitor-I (TAXI-I) has been shown to inhibit BcXyn11a, we verified if TAXI-I could be exploited to counteract B. cinerea infections. With this aim, we first produced Nicotiana tabacum plants transiently expressing TAXI-I, observing increased resistance to B. cinerea. Subsequently, we transformed Arabidopsis thaliana to express TAXI-I constitutively, and we obtained three transgenic lines exhibiting a variable amount of TAXI-I. The line with the higher level of TAXI-I showed increased resistance to B. cinerea and the absence of necrotic lesions when infiltrated with BcXyn11a. Finally, in a droplet application experiment on wild-type Arabidopsis leaves, TAXI-I prevented the necrotizing activity of BcXyn11a. These results would confirm that the contribution of BcXyn11a to virulence is due to its necrotizing rather than enzymatic activity. In conclusion, our experiments highlight the ability of the TAXI-I xylanase inhibitor to counteract B. cinerea infection presumably by preventing the necrotizing activity of BcXyn11a.

Microbial xylanases and their inhibition by specific proteins in cereals

Arabinoxylans (AXs) belong to the components of plant cells which are mainly degraded by microbial xylanases during colonization of grain by phytopathogens. For the defence, cereals contain proteinaceous xylanase inhibitors (XIs), namely xylanase inhibitor protein (XIP), Triticum aestivum xylanase inhibitor (TAXI) and thaumatin-like xylanase inhibitor (TLXI). Their presence in cereals in high levels can be a serious problem in different industrial applications. XIs regulate AX hydrolysis and participate in plant defence mechanisms. XIs have various specificity against microbial xylanases from the glycoside hydrolase (GH) families of GH10 and GH11. Therefore, this review brings new information about the function of XIs as defence responses to pathogen infection of plants and as a problem in plant material processing in different industrial applications.