Differentiation of xylem elements involves cell expansion, secondary cell wall (SCW) deposition and programmed cell death. Transitions between these phases require strict spatiotemporal control. The function of Populus ERF139 (Potri.013G101100) in xylem differentiation was characterized in transgenic overexpression and dominant repressor lines of ERF139 in hybrid aspen (Populus tremula × tremuloides). Xylem properties, SCW chemistry and downstream targets were analyzed in both types of transgenic trees using microscopy techniques, Fourier transform-infrared spectroscopy, pyrolysis-GC/MS, wet chemistry methods and RNA sequencing. Opposite phenotypes were observed in the secondary xylem vessel sizes and SCW chemistry in the two different types of transgenic trees, supporting the function of ERF139 in suppressing the radial expansion of vessel elements and stimulating accumulation of guaiacyl-type lignin and possibly also xylan. Comparative transcriptomics identified genes related to SCW biosynthesis (LAC5, LBD15, MYB86) and salt and drought stress-responsive genes (ANAC002, ABA1) as potential direct targets of ERF139. The phenotypes of the transgenic trees and the stem expression profiles of ERF139 potential target genes support the role of ERF139 as a transcriptional regulator of xylem cell expansion and SCW formation, possibly in response to osmotic changes of the cells.

https://nph.onlinelibrary.wiley.com/doi/pdf/10.1111/nph.15960

An AP2/ERF transcription factor ERF139 coordinates xylem cell expansion and secondary cell wall deposition.

Stem bending or lodging caused by insufficient stem strength is an important limiting factor for plant production. Secondary cell walls play a crucial role in plant stem strength, but whether WRKY transcription factors can positively modulate secondary cell wall thickness are remian unknown. Here, we characterized a WRKY transcription factor PlWRKY41a from herbaceous peony (Paeonia lactiflora), which was highly expressed in stems. PlWRKY41a functioned as a nucleus-localized transcriptional activator and enhanced stem strength by positively modulating secondary cell wall thickness. Moreover, PlWRKY41a bound to the promoter of the XYLOGLUCAN ENDOTRANSGLUCOSYLASE/HYDROLASE4 (PlXTH4) and activated the expression of PlXTH4. PlXTH4-overexpressing tobacco (Nicotiana tabacum) had thicker secondary cell walls, resulting in enhanced stem strength, while PlXTH4-silenced P. lactiflora had thinner secondary cell walls, showing decreased stem strength. Additionally, PlWRKY41a directly interacted with PlMYB43 to form a protein complex, and their interaction induced the expression of PlXTH4. These data support that the PlMYB43-PlWRKY41a protein complex can directly activate the expression of PlXTH4 to enhance stem strength by modulating secondary cell wall thickness in P. lactiflora. The results will enhance our understanding of the formation mechanism of stem strength and provide a candidate gene to improve stem straightness in plants.

The herbaceous peony transcription factor WRKY41a promotes secondary cell wall thickening to enhance stem strength.

The insufficient number of available simple sequence repeats (SSRs) inhibits genetic research on and molecular breeding of Paeonia lactiflora, a flowering crop with great economic value. The objective of this study was to develop SSRs for P. lactiflora with Illumina RNA sequencing and assess the role of SSRs in gene regulation. The results showed that dinucleotides with AG/CT repeats were the most abundant type of repeat motif in P. lactiflora and were preferentially distributed in untranslated regions. Significant differences in SSR size were observed among motif types and locations. A large number of unigenes containing SSRs participated in catalytic activity, metabolic processes and cellular processes, and 28.16% of all transcription factors and 21.74% of hub genes for inflorescence stem straightness were found to contain SSRs. Successful amplification was achieved with 89.05% of 960 pairs of SSR primers, 55.83% of which were polymorphic, and most of the 46 tested primers had a high level of transferability to the genus Paeonia. Principal component and cluster dendrogram analyses produced results consistent with known genealogical relationships. This study provides a set of SSRs with abundant information for future accession identification, marker-trait association and molecular assisted breeding in P. lactiflora.

Characteristics of Microsatellites Mined from Transcriptome Data and the Development of Novel Markers in Paeonia lactiflora

Gerbera, one of the most loved cut flowers, is (in)famous for the drooping of its flower heads under dehydration. This effect has been quantified by analysing both fully turgescent and wilting peduncles of Gerbera jamesonii 'Nuance'. Wilting peduncles display pronounced bending in the region directly below the inflorescence after 24 h of dehydration, while the rest of the peduncle remains upright. Using anatomical measurements and three-point bending tests, we have analysed whether this phenomenon is caused by mechanical and/or geometrical alterations. We have found that both the flexural rigidity and the axial second moment of area are significantly decreased in the apical part of wilting peduncles, whereas the bending elastic modulus shows no significant change. Moreover, cross-sections of wilting peduncles ovalize significantly more than those of turgescent peduncles and exhibit considerable shrinkage of the parenchyma, taking up the majority of the cross-sectional area. Generally, the drooping of wilting Gerbera flowers can be regarded as a temporary instability of a rod-shaped cellular solid caused by anatomical differences (tissue arrangement, existence or the absence of a pith cavity) and geometrical changes (the decrease of axial second moment of area, cross-sectional ovalization, shrinkage of tissues) between the apical and basal regions of their peduncles.

https://royalsocietypublishing.org/doi/pdf/10.1098/rsbl.2019.0254

Drooping of Gerbera flower heads: mechanical and structural studies of a well-known phenomenon.

Paeonia emodi is a type of wild herbaceous peony with high ornamental and breeding value. Cold stratification is the only method to break its seed epicotyl dormancy to date, however, the key physiological factors during this process are not clear. In this study, rooted seeds of P. emodi were treated with 4 °C stratification, and the changes of the embryo, four nutrients, and two endogenous hormones in the seeds were investigated. The results showed that the plumule elongated at S6 (i.e., ten weeks of cold stratification), and grew to nearly the same length as the cotyledon at S9. Cold stratification increased starch consumption, significantly decreased soluble sugar content in the later stages, and decreased soluble protein content at S9, but it did not influence crude fat content. The activities of α-amylase and β-amylase increased significantly at S4 and S4 to S6, respectively. At S8 and S9, acid protease activity increased, and the increase in lipase activity continued throughout the whole process. At the same time, the ABA content decreased significantly after S6; from this stage, the ABA/GA3 ratio gradually decreased compared with that of the control, and the difference was significant at S9. Correlation analysis showed that the ABA/GA3 ratio was significantly correlated with starch content and α-amylase activity. It can be concluded that both carbohydrates and proteins were the energy supply for the epicotyl dormancy breaking of P. emodi seeds, rather than crude fat. Cold stratification promoted substance transformation by increasing the corresponding enzyme activities. The balance of ABA and GA3 suggested the key stage for the release of dormancy.

/pdf/morphology-and-physiology-response-to-stratification-during-27rrkzhp.pdf

Morphology and Physiology Response to Stratification during Seeds Epicotyl Dormancy Breaking of Paeonia emodi Wall. ex Royle

Lack of structural components results in inflorescence stem bending. Differentially expressed genes involved in lignin and hemicellulose biosynthesis are vital; genes involved in cellulose and glycan biosynthesis are also relevant. An erect inflorescence stem is essential for high-quality cut herbaceous peony flowers. To explore the factors underlying inflorescence stem bending, major cell walls contents were measured, and stem structure was observed in two herbaceous peony varieties with contrasting stem straightness traits (‘Da Fugui’, upright; ‘Chui Touhong’, bending). In addition, Illumina sequencing was performed and weighted correlation network analysis (WGCNA) was used to analyze the results. The results showed significant differences in lignin, hemicellulose and soluble sugar contents, sclerenchyma and xylem areas and thickening in cell walls in pith at stage S3, when bending begins. In addition, 44,182 significantly differentially expressed genes (DEGs) were found, and these DEGs were mainly enriched in 36 pathways. Among the DEGs, hub genes involved in lignin, cellulose, and xylan biosynthesis and transcription factors that regulated these process were identified by WGCNA. These results suggested that the contents of compounds that provided cell wall rigidity were vital factors affecting inflorescence stem straightness in herbaceous peony. Genes involved in or regulating the biosynthesis of these compounds are thus important; lignin and hemicellulose are of great interest, and cellulose and glycan should not be ignored. This paper lays a foundation for developing new herbaceous peony varieties suitable for cut flowers by molecular-assisted breeding.

Transcriptome and weighted correlation network analyses provide insights into inflorescence stem straightness in Paeonia lactiflora.

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Transcriptome and weighted correlation network analyses provide insights into inflorescence stem straightness in Paeonia lactiflora.