Identification and functional analysis of arabinogalactan protein expressed in pear pollen tubes

Arabinogalactan proteins(AGPs)are widely distributed in the plant kingdom and play a vital role during the process of plant sexual reproduction.In this study,we performed a comprehensive identification of the PbrAGPs expressed in pear pollen and further explored their influences on pollen tube growth.Among the 187 PbrAGPs that were found to be expressed in pear pollen tubes,38 PbrAGPs were specifically expressed in pollen according to the RNA-seq data.The PbrAGPs were divided into two groups of highly expressed and specifically expressed in pear pollen.We further tested their expression patterns using RT-PCR and RT-qPCR.Most of the PbrAGPs were expressed in multiple tissues and their expression levels were consistent with reads per kilobase per million map reads(RPKM)values during pollen tube growth,implying that PbrAGPs might be involved in the regulation of pear pollen tube growth.We also constructed phylogenetic trees to identify the functional genes in pear pollen tube growth.Therefore,19 PbrAGPs(PbrAGP1 to PbrAGP19)were selected to test their influences on pollen tube growth.Recombinant proteins of the 19 PbrAGP-His were purified and used to treat pear pollen,and 11 of the PbrAGP-His recombinant proteins could promote pear pollen tube growth.Additionally,pollen tube growth was inhibited when the expression levels of PbrAGP1 and PbrAGP5 were knocked down using an antisense oligonucleotide assay.PbrAGP1 and PbrAGP5 were localized in the plasma membrane and might not alter the distribution of pectin in the pollen tube.In summary,this study identified the PbrAGPs expressed in pear pollen and lays the foundation for further exploring their functions in pollen tube growth.

DEAP1 encodes a fasciclin-like arabinogalactan protein required for male fertility in rice

Arabinogalactan proteins(AGPs)are widely distributed in plant cells.Fasciclin-like AGPs(FLAs)belong to a subclass of AGPs that play important roles in plant growth and development.However,little is known about the biological functions of rice FLA.Herein,we report the identification of a male-sterile mutant of DEFECTIVE EXINE AND APERTURE PATTERNING1(DEAP1)in rice.The deap1 mutant anthers produced aberrant pollen grains with defective exine formation and a flattened aperture annulus and exhibited slightly delayed tapetum degradation.DEAP1 encodes a plasma membrane-associated member of groupⅢplant FLAs and is specifically and temporally expressed in reproductive cells and the tapetum layer during male development.Gene expression studies revealed reduced transcript accumulation of genes related to exine formation,aperture patterning,and tapetum development in deap1 mutants.Moreover,DEAP1 may interact with two rice D6 PROTEIN KINASE-LIKE3 s(OsD6PKL3s),homologs of a known Arabidopsis aperture protein,to affect rice pollen aperture development.Our findings suggested that DEAP1 is involved in male reproductive development and may affect exine formation and aperture patterning,thereby providing new insights into the molecular functions of plant FLAs in male fertility.

Enrichment process of lanthanum as a nonessential trace element in leaf cells of lettuce(Lactuca sativa L.)

Rare earth elements(REEs)as nonessential trace elements are enriched in living organisms and threaten their health.To early detect and reduce REE enrichment in living organisms,scientists are focused on clarifying the enrichment process of REEs in living organisms and its risks.However,the enrichment process of REEs in edible plant cells has remained unclear.Herein,by using interdisciplinary methods and techniques,the enrichment process of lanthanum(La(Ⅲ))in the leaf cells of lettuce(Lactuca sativa L.)was investigated.(1)When La(Ⅲ)exposure dose is 0.5-5μmol/L,La(Ⅲ)is enriched outside the plasma membrane(PM).In this zone,La(Ⅲ)is bound to vitronectin-like protein(VN)to form La-VN complexes;(2)When La(Ⅲ)exposure dose is 5-20μmol/L,besides the zone outside the PM,La(Ⅲ)is also enriched on the PM and bound to arabinogalactan proteins(AGPs)to form La-AGPs complexes;(3)When La(Ⅲ)exposure dose is 20-140μmol/L,besides the zone outside and on the PM,La(Ⅲ)is enriched inside the PM;(4)When La(Ⅲ)exposure dose is 60-140μmol/L,malondialdehyde content(an important indicator of invisible damage)significantly increases.Thus,as La(Ⅲ)exposure dose increases,La(Ⅲ)gradually migrates from outside the PM to the PM and inside the PM,enriching in these zones in turn.The enriched La(Ⅲ)will cause invisible damage to lettuce leaf cells and even enter human bodies along food chains.These results provide references for investigating the enrichment process of REEs in plants and its environmental risks,and finding strategies to early detect and reduce REE enrichment in plants.

BRITTLE PLANT1 is required for normal cell wall composition and mechanical strength in rice

A series of nucleotide sugar interconversion enzymes(NSEs) generate the activated sugar donors required for biosynthesis of cell wall matrix polysaccharides and glycoproteins. UDPglucose 4-epimerases(UGEs) are NSEs that function in the interconversion of UDP-glucose(UDP-Glc) and UDP-galactose(UDP-Gal). The roles of UDP-glucose 4-epimerases in monocots remain unclear due to redundancy in the pathways. Here, we report a brittle plant(bp1) rice mutant that exhibits brittle leaves and culms at all growth stages. The mutant culms had reduced levels of rhamnogalacturonan I, homogalacturonan, and arabinogalactan proteins.Moreover, the mutant had altered contents of uronic acids, neutral noncellulosic monosaccharides, and cellulose. Map-based cloning demonstrated that OsBP1 encodes a UDPglucose 4-epimerase(OsUGE2), a cytosolic protein. We also show that BP1 can form homoand hetero-protein complexes with other UGE family members and with UDP-galactose transporters 2(OsUGT2) and 3(OsUGT3), which may facilitate the channeling of Gal to polysaccharides and proteoglycans. Our results demonstrate that BP1 participates in regulating the sugar composition and structure of rice cell walls.