Genome-wide identification of the pectate lyase(PEL)gene family members in Malvaceae,and their contribution to cotton fiber quality

Pectin is a major constituent of the plant cell wall.Pectate lyase(PEL,EC 4.2.2.2)uses anti-β-elimination chemistry to cleave theα-1,4 glycosidic linkage in the homogalacturonan region of pectin.However,limited information is available on the comprehensive and evolutionary analysis of PELs in the Malvaceae.In this study,we identified 597PEL genes from 10 Malvaceae species.Phylogenetic and motif analyses revealed that these PELs are classified into six subfamilies:Clades I,II,III,IV,Va,and Vb.The two largest subfamilies,Clades I and II,contained 237 and222 PEL members,respectively.The members of Clades Va and Vb only contained four or five motifs,far fewer than the other subfamilies.Gene duplication analysis showed that segmental duplication played a crucial role in the expansion of the PEL gene family in Gossypium species.The PELs from Clades I,IV,Va,and Vb were expressed during the fiber elongation stage,but nearly all PEL genes from Clades II and III showed no expression in any of the investigated fiber developmental stages.We further performed single-gene haplotype association analysis in 2,001G.hirsutum accessions and 229 G.barbadense accessions.Interestingly,14 PELs were significantly associated with fiber length and strength traits in G.barbadense with superior fiber quality,while only eight GhPEL genes were found to be significantly associated with fiber quality traits in G.hirsutum.Our findings provide important information for further evolutionary and functional research on the PEL gene family members and their potential use for fiber quality improvement in cotton.

Pectate lyase is a factor in the adaptability for Heterodera glycines infecting tobacco

The soybean cyst nematode, Heterodeara glycines, is a serious pathogen of soybean, and reported to be the host of a wide range of Fabaceae. In the present study, the host specificity and reproductivity of two populations of H. glycines collected from soybean and tobacco were identified and characterized. The comparative identity between β-1,4-endoglucanase, pectate lyase and chorismate mutase of H. glycines parasitizing on soybean and tobacco were 99, 97 and 98%, respectively. The qR T-PCR analysis indicated that the expression of pectate lyase 2 gene was significantly higher in second-stage juveniles of H. glycines Henan population parasitizing on tobacco than that of H. glycines Shanxi population parasitizing on soybean. In addition, the pectic acid content of cell wall was significantly higher(45%) in the roots of tobacco than the roots of soybean. Our results indicate that the changes in transcript parasitism genes may be a result of long-term evolution illustrating how a plant-parasitic nematode adapts to the host environment for optimal infestation and survival.

Bioscouring Knitted Cotton Fabric with an Experimental Pectate Lyase

An experimental pectate lyase enzyme was used to scour knitted cotton fabric and the emphasis was on pectin removal.Using an enzyme dosage of 0.2 g/L at temperature 55℃ and pH 6.35 for 30 min,good scouring properties were obtained.When appropriate concentrations of 1-Hydroxy Ethylidene-1,1-Diphosphonic Acid(HEDP)and CaCl2 were added,the percentage pectin removal improved significantly.

PECTATE LYASE-LIKE10 is associated with pollen wall development in Brassica campestris

PECTATE LYASE‐LIKE10（PLL10） was previously identified as one of the differentially expressed genes both in microspores during the late pollen developmental stages and in pistils during the fertilization process in Chinese cabbage（Brassica campestris ssp. chinensis）. Here, antisense‐RNA was used to study the functions of BcPLL10 in Chinese cabbage. Abnormal pollen was identified in the transgenic lines（bcpll10‐4, ‐5, and ‐6）. In fertilization experiments, fewer seeds were harvested when the antisense‐RNA lines were used as pollen donor. In vivo and in vitro pollen germination assays less germinated pollen tubes were observed in bcpll10 lines. Scanning electron microscopy observation verified that the tryphine materials were over accumulated around the pollen surface and sticked them together in bcpll10.Moreover, transmission electron microscopy observation revealed that the internal endintine was overdeveloped and predominantly occupied the intine, and disturbed thenormal proportional distribution of the two layers in the non‐germinal furrow region; and no obvious demarcation existed between them in the germinal furrow region in the bcpll10 pollen. Collectively, this study presented a novel PLL gene that played an important role during the pollen wall development in B. campestris, which may also possess potential importance for male sterility usage in agriculture.

Locus TUTOU2 determines the panicle apical abortion phenotype of rice(Oryza sativa L.)in tutou2 mutant

Rice panicle apical abortion(PAA)is a detrimental agronomic trait resulting in spikelet number reduction and yield loss.To understand its underlying molecular mechanism,we identified one recessive PAA mutant tutou2 from the offspring of tissue cultures.The mutation locus was finely mapped to a 75-kb interval on the long arm of chromosome 10.Sequence analysis revealed a single nucleotide substitution of A to T at the 941 position of LOC_Os10g31910 in tutou2,resulting in an amino acid change from isoleucine to phenylalanine.Complementation analysis showed that the degenerated panicle phenotype in tutou2 was rescued in the transgenic lines.A phenotype similar to tutou2 can also be obtained by LOC_Os10g31910 knockout in wild-type rice.These results suggested that LOC_Os10 g31910 is the causative locus TUTOU2 responsible for the tutou2 PAA phenotype and probably also the locus of DEL1,previously documented as a leaf senescence gene.The significant phenotypic differences between del1 and tutou2 suggest that the locus DEL1/TUTOU2 plays roles in both leaf and panicle development which were not considered fully in previous studies.

Production of Pectinolytic Enzymes by Two Bacillus spp. Strains and Their Application in Flax Degumming

This study demonstrated that Bacillus licheniformis HDYM-03 and Bacillus megaterium HDYM-09, isolated from a liquid sample of fl ax retting pool, were able to produce pectinolytic enzymes using polysaccharides as substrates. Bacillus megaterium HDYM-09 produced pectin lyase that exhibited the highest activity of 2116.71 ± 11.55 U/mL. Bacillus licheniformis HDYM-03 produced pectate lyase that exhibited the highest activity of 611.21 ± 14.54 U/mL. Based on these fi ndings, we constructed four retting systems to degrade the pectin substance. The results showed that the content of galacturonic acid in the mixed system was 529.21 μg/mL, the content of reducing sugar was 98.14 mg/mL, and the weight loss ratio of cells reached 19.49%, which were signifi cantly higher than those in other systems. The mixed system has more advantages, and the utilization rate of degumming was higher, which further ensured that the degumming can be carried out effi ciently and quickly. The mixed system exhibits feasible applications in the fi ber and textile industry.

Combinatorial Enzyme Approach to Produce Oligosaccharides of Diverse Structures for Functional Screen

Combinatorial chemistry has been a focus of research activity in modern drug discovery and biotechnology. It is a concept by which a vast library of molecular diversity is synthesized and screened for target properties. This report is to illustrate the application of enzyme technology using the concept of combinatorial chemistry as a novel approach for the bioconversion of plant fibers. Citrus pectin was subjected to combinatorial enzyme digestion to create libraries of pectic oligosaccharides with diverse structural variants. Repeated cycles of fractionation and screening resulted in the isolation and identification of an active oligoGalA species with antimicrobial activity.

Ethylene†and fruit softening

This review is concerned with the mechanisms controlling fruit softening.Master genetic regulators switch on the ripening programme and the regulatory pathway branches downstream,with separate controls for distinct quality attributes such as colour,flavour,texture,and aroma.Ethylene plays a critical role as a ripening hormone and is implicated in controlling different facets of ripening,including texture change,acting through a range of transcriptional regulators,and this signalling can be blocked using 1-methylcyclopropene.A battery of at least seven cell-wall-modifying enzymes,most of which are synthesized de novo during ripening,cause major alterations in the structure and composition of the cell wall components and contribute to the softening process.Significant differences between fruits may be related to the precise structure and composition of their cell walls and the enzymes recruited to the ripening programme during evolution.Attempts to slow texture change and reduce fruit spoilage by delaying the entire ripening process can often affect negatively other aspects of quality,and low temperatures,in particular,can have deleterious effects on texture change.Gene silencing has been used to probe the function of individual genes involved in different aspects of ripening,including colour,flavour,ethylene synthesis,and particularly texture change.The picture that emerges is that softening is a multi-genic trait,with some genes making a more important contribution than others.In future,it may be possible to control texture genetically to produce fruits more suitable for our needs.