Defying gravity: WEEP promotes negative gravitropism in peach trees by establishing asymmetric auxin gradients.

Trees with weeping shoot architectures are valued for their beauty and are a resource for understanding how plants regulate posture control. The peach (Prunus persica) weeping phenotype, which has elliptical downward arching branches, is caused by a homozygous mutation in the WEEP gene. Little is known about the function of WEEP despite its high conservation throughout Plantae. Here, we present the results of anatomical, biochemical, biomechanical, physiological, and molecular experiments that provide insight into WEEP function. Our data suggest that weeping peach trees do not have defects in branch structure. Rather, transcriptomes from the adaxial (upper) and abaxial (lower) sides of standard and weeping branch shoot tips revealed flipped expression patterns for genes associated with early auxin response, tissue patterning, cell elongation, and tension wood development. This suggests that WEEP promotes polar auxin transport toward the lower side during shoot gravitropic response, leading to cell elongation and tension wood development. In addition, weeping peach trees exhibited steeper root systems and faster lateral root gravitropic response. This suggests that WEEP moderates root gravitropism and is essential to establishing the set-point angle of lateral roots from the gravity vector. Additionally, size exclusion chromatography indicated that WEEP proteins self-oligomerize, like other proteins with sterile alpha motif domains. Collectively, our results from weeping peach provide insight into polar auxin transport mechanisms associated with gravitropism and lateral shoot and root orientation.

The 141-year period for Dr. Beal's seed viability experiment: A hybrid surprise.

PREMISE: In 1879, Dr. William Beal buried 20 glass bottles filled with seeds and sand at a single site at Michigan State University. The goal of the experiment was to understand seed longevity in the soil, a topic of general importance in ecology, restoration, conservation, and agriculture, by periodically assaying germinability of these seeds over 100 years. The interval between germination assays has been extended and the experiment will now end after 221 years, in 2100. METHODS: We dug up the 16th bottle in April 2021 and attempted to germinate the 141-year-old seeds it contained. We grew germinants to maturity and identified these to species by vegetative and reproductive phenotypes. For the first time in the history of this experiment, genomic DNA was sequenced to confirm species identities. RESULTS: Twenty seeds germinated over the 244-day assay. Eight germinated in the first 11 days. All 20 belonged to the Verbascum genus: Nineteen were V. blattaria according to phenotype and ITS2 genotype; and one had a hybrid V. blattaria × V. thapsus phenotype and ITS2 genotype. In total, 20/50 (40%) of the original Verbascum seeds in the bottle germinated in year 141. CONCLUSIONS: While most species in the Beal experiment lost all seed viability in the first 60 years, a high percentage of Verbascum seeds can still germinate after 141 years in the soil. Long-term experiments such as this one are rare and invaluable for studying seed viability in natural soil conditions.

Defying Gravity: WEEP promotes negative gravitropism in Prunus persica (peach) shoots and roots by establishing asymmetric auxin gradients.

Trees with weeping shoot architectures are valued for their beauty and serve as tremendous resources for understanding how plants regulate posture control. The Prunus persica (peach) weeping phenotype, which has elliptical downward arching branches, is caused by a homozygous mutation in the WEEP gene. Until now, little was known about the function of WEEP protein despite its high conservation throughout Plantae. Here, we present the results of anatomical, biochemical, biomechanical, physiological, and molecular experiments that provide insight into WEEP function. Our data suggest that weeping peach does not have defects in branch structure. Rather, transcriptomes from the adaxial (upper) and abaxial (lower) sides of standard and weeping branch shoot tips revealed flipped expression patterns for genes associated with early auxin response, tissue patterning, cell elongation, and tension wood development. This suggests that WEEP promotes polar auxin transport toward the lower side during shoot gravitropic response, leading to cell elongation and tension wood development. In addition, weeping peach trees exhibited steeper root systems and faster root gravitropic response, just as barley and wheat with mutations in their WEEP homolog EGT2. This suggests that the role of WEEP in regulating lateral organ angles and orientations during gravitropism may be conserved. Additionally, size-exclusion chromatography indicated that WEEP proteins self-oligomerize, like other SAM-domain proteins. This oligomerization may be required for WEEP to function in formation of protein complexes during auxin transport. Collectively, our results from weeping peach provide new insight into polar auxin transport mechanisms associated with gravitropism and lateral shoot and root orientation.

Environmental-biomechanical reciprocity and the evolution of plant material properties.

Abiotic-biotic interactions have shaped organic evolution since life first began. Abiotic factors influence growth, survival, and reproductive success, whereas biotic responses to abiotic factors have changed the physical environment (and indeed created new environments). This reciprocity is well illustrated by land plants who begin and end their existence in the same location while growing in size over the course of years or even millennia, during which environment factors change over many orders of magnitude. A biomechanical, ecological, and evolutionary perspective reveals that plants are (i) composed of materials (cells and tissues) that function as cellular solids (i.e. materials composed of one or more solid and fluid phases); (ii) that have evolved greater rigidity (as a consequence of chemical and structural changes in their solid phases); (iii) allowing for increases in body size and (iv) permitting acclimation to more physiologically and ecologically diverse and challenging habitats; which (v) have profoundly altered biotic as well as abiotic environmental factors (e.g. the creation of soils, carbon sequestration, and water cycles). A critical component of this evolutionary innovation is the extent to which mechanical perturbations have shaped plant form and function and how form and function have shaped ecological dynamics over the course of evolution.

Pinus ponderosa: A checkered past obscured four species.

PREMISE OF THE STUDY: Molecular genetic evidence can help delineate taxa in species complexes that lack diagnostic morphological characters. Pinus ponderosa (Pinaceae; subsection Ponderosae) is recognized as a problematic taxon: plastid phylogenies of exemplars were paraphyletic, and mitochondrial phylogeography suggested at least four subdivisions of P. ponderosa. These patterns have not been examined in the context of other Ponderosae species. We hypothesized that putative intraspecific subdivisions might each represent a separate taxon. METHODS: We genotyped six highly variable plastid simple sequence repeats in 1903 individuals from 88 populations of P. ponderosa and related Ponderosae (P. arizonica, P. engelmannii, and P. jeffreyi). We used multilocus haplotype networks and discriminant analysis of principal components to test clustering of individuals into genetically and geographically meaningful taxonomic units. KEY RESULTS: There are at least four distinct plastid clusters within P. ponderosa that roughly correspond to the geographic distribution of mitochondrial haplotypes. Some geographic regions have intermixed plastid lineages, and some mitochondrial and plastid boundaries do not coincide. Based on relative distances to other species of Ponderosae, these clusters diagnose four distinct taxa. CONCLUSIONS: Newly revealed geographic boundaries of four distinct taxa (P. benthamiana, P. brachyptera, P. scopulorum, and a narrowed concept of P. ponderosa) do not correspond completely with taxonomies. Further research is needed to understand their morphological and nuclear genetic makeup, but we suggest that resurrecting originally published species names would more appropriately reflect the taxonomy of this checkered classification than their current treatment as varieties of P. ponderosa.

Molecular cloning of the tomato Hairless gene implicates actin dynamics in trichome-mediated defense and mechanical properties of stem tissue.

Trichomes are epidermal structures that provide a first line of defense against arthropod herbivores. The recessive hairless (hl) mutation in tomato (Solanum lycopersicum L.) causes severe distortion of trichomes on all aerial tissues, impairs the accumulation of sesquiterpene and polyphenolic compounds in glandular trichomes, and compromises resistance to the specialist herbivore Manduca sexta Here, we demonstrate that the tomato Hl gene encodes a subunit (SRA1) of the highly conserved WAVE regulatory complex that controls nucleation of actin filaments in a wide range of eukaryotic cells. The tomato SRA1 gene spans a 42-kb region containing both Solyc11g013280 and Solyc11g013290 The hl mutation corresponds to a complex 3-kb deletion that removes the last exon of the gene. Expression of a wild-type SRA1 cDNA in the hl mutant background restored normal trichome development, accumulation of glandular trichome-derived metabolites, and resistance to insect herbivory. These findings establish a role for SRA1 in the development of tomato trichomes and also implicate the actin-cytoskeleton network in cytosolic control of specialized metabolism for plant defense. We also show that the brittleness of hl mutant stems is associated with altered mechanical and cell morphological properties of stem tissue, and demonstrate that this defect is directly linked to the mutation in SRA1.

Comment on "Critical wind speed at which trees break".

Virot et al. [E. Virot et al., Phys. Rev. E 93, 023001 (2016)10.1103/PhysRevE.93.023001] assert that the critical wind speed at which ⩾50% of all trees in a population break is ≈42 m/s, regardless of tree characteristics. We show that empirical data do not support this assertion, and that the assumptions underlying the theory used by Virot et al. are inconsistent with the biomechanics of trees.

Acclimation of mechanical and hydraulic functions in trees: impact of the thigmomorphogenetic process.

The secondary xylem (wood) of trees mediates several functions including water transport and storage, mechanical support and storage of photosynthates. The optimal structures for each of these functions will most likely differ. The complex structure and function of xylem could lead to trade-offs between conductive efficiency, resistance to embolism, and mechanical strength needed to count for mechanical loading due to gravity and wind. This has been referred to as the trade-off triangle, with the different optimal solutions to the structure/function problems depending on the environmental constraints as well as taxonomic histories. Thus, the optimisation of each function will lead to drastically different anatomical structures. Trees are able to acclimate the internal structure of their trunk and branches according to the stress they experience. These acclimations lead to specific structures that favor the efficiency or the safety of one function but can be antagonistic with other functions. Currently, there are no means to predict the way a tree will acclimate or optimize its internal structure in support of its various functions under differing environmental conditions. In this review, we will focus on the acclimation of xylem anatomy and its resulting mechanical and hydraulic functions to recurrent mechanical strain that usually result from wind-induced thigmomorphogenesis with a special focus on the construction cost and the possible trade-off between wood functions.

Chemical responses to modified lignin composition in tension wood of hybrid poplar (Populus tremula x Populus alba).

The effect of altering the expression level of the F5H gene was investigated in three wood tissues (normal, opposite and tension wood) in 1-year-old hybrid poplar clone 717 (Populus tremula × Populus alba L.), containing the F5H gene under the control of the C4H promoter. Elevated expression of the F5H gene in poplar has been previously reported to increase the percent syringyl content of lignin. The wild-type and three transgenic lines were inclined 45° for 3 months to induce tension wood formation. Tension and opposite wood from inclined trees, along with normal wood from control trees, were analyzed separately for carbohydrates, lignin, cellulose crystallinity and microfibril angle (MFA). In the wild-type poplar, the lignin in tension wood contained a significantly higher percentage of syringyl than normal wood or opposite wood. However, there was no significant difference in the percent syringyl content of the three wood types within each of the transgenic lines. Increasing the F5H gene expression caused an increase in the percent syringyl content and a slight decrease in the total lignin in normal wood. In tension wood, the addition of a gelatinous layer in the fiber walls resulted in a consistently lower percentage of total lignin in the tissue. Acid-soluble lignin was observed to increase by up to 2.3-fold in the transgenic lines. Compared with normal wood and opposite wood, cell wall crystallinity in tension wood was higher and the MFA was smaller, as expected, with no evidence of an effect from modifying the syringyl monomer ratio. Tension wood in all the lines contained consistently higher total sugar and glucose percentages when compared with normal wood within the respective lines. However, both sugar and glucose percentages were lower in the tension wood of transgenic lines when compared with the tension wood of wild-type trees. Evaluating the response of trees with altered syringyl content to gravity will improve our understanding of the changes in cell wall chemistry and ultrastructural properties of normal, opposite and tension wood in plants.

Importance of two consecutive methionines at the N-terminus of a cellulose synthase (PtdCesA8A) for normal wood cellulose synthesis in aspen.

All known orthologs of a secondary wall-associated cellulose synthase (CesA) gene from Arabidopsis, AtCesA8, encode CesA proteins with two consecutive methionines at their N-termini (MM or 2M). Here, we report that these 2Ms in an aspen ortholog of AtCesA8, PtdCesA8A, are important for maintaining normal wood cellulose biosynthesis in aspen trees. Overexpression of an altered PtdCesA8A cDNA encoding a PtdCesA8A protein missing one methionine at the N-terminus (1M) in aspen resulted in substantial decrease in cellulose content and caused negative effects on wood strength, suggesting that both methionines are essential for proper CesA expression and function in developing xylem tissues. Transcripts from a pair of paralogous native PtdCesA8 genes, as well as introduced PtdCesA8A:1M transgenes were significantly reduced in developing xylem tissues of transgenic aspen plants, suggestive of a co-suppression event. Overexpression of a native PtdCesA8A cDNA encoding a CesA protein with 2Ms at the N-terminus did not cause any such phenotypic changes. These results suggest the importance of 2Ms present at the N-terminus of PtdCesA8A protein during cellulose synthesis in aspen.

Is windswept tree growth negative thigmotropism?

The scientific investigation of the influence of wind on tree growth has been conducted for over 200 years. One influence of wind on trees is the formation of an asymmetric crown, usually characterized as being windswept under moderate windy conditions. As wind exposure increases, the terms applied to this growth form include flag-tree, banner-tree, and krummholz. The modification in crown morphology has been widely recognized and studied, especial in the area of wind prospecting or as a bioindicator of wind speed in environments lacking monitoring stations. However, the causes and physiology underlying this response is little understood. The windswept morphology is consistent with the morphologies associated with other tropisms (i.e. phototropism and gravitropism). Tropisms are defined as a growth response towards (positive) or away (negative) from an environmental stimulus. The asymmetric growth form of windswept trees appears to be a negative thigmotropic growth response to wind. In this review, evidence will be presented to support or reject two hypotheses; H1 the windswept growth form is the result of a negative thigmotropic growth response or H2 the windswept growth form is determined by the biophysical properties of wood. It is argued that the windswept growth form is more likely the product of biomechanical properties (accept H2) than of a physiological thigmotropic growth response (reject H1). However, proper testing of both hypotheses is still required before a final confirmation can be established.

Chloroplast diversity in a putative hybrid swarm of Ponderosae (Pinaceae).

The Ponderosae subsection of the genus Pinus contains numerous taxa in disjunct mountain ranges of southern Arizona and New Mexico, differing for several leaf and cone traits, key among which is the number of leaf needles per fascicle. Trees with three needles are often found together with trees having five needles and mixed numbers. One taxonomic hypothesis is that there are swarms of hybrids between P. ponderosa and P. arizonica. A second hypothesis is that there are spatial mixtures of two separate taxa, five-needle P. arizonica and a "taxon X" containing three needle and mixed needle trees. We genotyped chloroplasts in one putative hybrid swarm on Mt. Lemmon using microsatellite markers and show that cpDNA is almost completely differentiated between two separate morphotypes corresponding to P. arizonica and "taxon X." Little if any introgression has occurred on Mt. Lemmon, and the simplest explanation is that little or no effective hybridization has occurred. Further results indicate that not only is taxon X not of hybrid origin, it is more closely related to nonregional Ponderosae other than P. ponderosa and P. arizonica. The results further suggest that other putative hybrid swarms in the region are also spatial mixtures of distinct taxa.

Biomechanics and transgenic wood.

Wood, or secondary xylem, is composed mostly of three components-cellulose, hemicelluloses, and lignin. Yet this apparent simplicity is deceiving because the sophisticated arrangement of the components on various structural levels, ranging from intricate molecular architecture to defined cellular arrangements to tissue morphology, makes wood a challenging and interesting subject of biomechanical investigation. Recent advances in genetic transformation, providing easier access to wood of specifically altered composition or structure, have opened new opportunities for research on the intricate relation between material structure and composition and mechanical properties. At the same time, investigations into the mechanical properties have provided new information regarding the structural configuration of wood. The present paper reviews the work conducted in this field and outlines future perspectives and prospects for research.

A unified hypothesis of mechanoperception in plants.

The perception of mechanical stimuli in the environment is crucial to the survival of all living organisms. Recent advances have led to the proposal of a plant-specific mechanosensory network within plant cells that is similar to the previously described network in animal systems. This sensory network is the basis for a unifying hypothesis, which may account for the perception of numerous mechanical signals including gravitropic, thigmomorphic, thigmotropic, self-loading, growth strains, turgor pressure, xylem pressure potential, and sound. The current state of our knowledge of a mechanosensory network in plants is reviewed, and two mechanoreceptor models are considered: a plasmodesmata-based cytoskeleton-plasma membrane-cell wall (CPMCW) network vs. stretch-activated ion channels. Post-mechanosensory physiological responses to mechanical stresses are also reviewed, and future research directions in the area of mechanoperception and response are recommended.

Mechanical perturbation affects conductivity, mechanical properties and aboveground biomass of hybrid poplars.

Xylem development in trees is affected by dynamic mechanical stresses imposed on stems by wind. To assess clonal differences in response to mechanical perturbation (MP), we subjected seven greenhouse-grown F1 hybrids of Populus trichocarpa Torr. and A. Gray. x P. deltoides Bartr. ex Marsh. to a standard MP treatment consisting of 20 manually imposed stem flexures per day for 70-90 days. Effects of MP on aboveground biomass, hydraulic conductivity (k(h)), specific conductivity (k(s)), flexural stiffness (EI), modulus of elasticity (MOE) and modulus of rupture (MOR) were determined. Treatment increased stem radial growth and decreased height growth, leaf area and total aboveground biomass. It also significantly decreased k(s), MOE and MOR, but significantly increased EI and wood specific gravity in most clones. Mechanical perturbation caused greater stem rigidity, without having a significant effect on whole-stem k(h) or percent loss of conductivity due to embolism. Maximum k(h) was positively correlated with EI in both control (r(2) = 0.54, P < 0.0001) and MP-treated (r(2) = 0.61, P < 0.0001) plants, and k(s) and MOE were positively correlated with percent vessel lumen area (r(2) = 0.45, P < 0.0001 and r(2) = 0.28, P = 0.002, respectively). Thus, contrary to our expectation of a trade-off between conductivity and wood strength, there may be an opportunity to select clones for woody biomass production that are superior in both mechanical strength and hydraulic conductivity, as is the triploid Clone 19-61.

Spatial pattern of allozyme variation in a contact zone of Pinus Ponderosa and P. arizonica (Pinaceae).

The spatial distribution of genotypes for nine polymorphic allozyme loci was examined in a contact zone between Pinus ponderosa var. scopulorum and another tree regarded as either a separate species, Pinus arizonica, or variety, Pinus ponderosa var. arizonica, in southern Arizona. Previous work had identified a steep elevational cline for a key taxonomic trait, number of leaf-needles per fascicle, on the south slope of Mt. Lemmon. The present results indicate that the taxa are not fully interbreeding in this contact zone, because allozyme genotypes are considerably more spatially structured than expected for the dispersal characteristics of pines. The amount of spatial differentiation is also much less than that observed for needle number. It appears that this is due to the lack of differentiation for allozyme gene frequencies for the two types of trees, which is further evidenced by analysis of samples from two other populations away from the contact zone. It is likely that if the two taxa were isolated in the past, it was not for long enough nor complete enough to allow mutation-drift to create substantial differentiation between them. Another possible explanation is that introgression after recontact is so advanced that any differences have been erased throughout the Santa Catalina mountain range.

Hydraulic, biomechanical, and anatomical interactions of xylem from five species of Acer (Aceraceae).

Possible trade-offs between hydraulic conductivity and mechanical properties of woody stems from five species were assessed. Acer negundo is a ruderal tree, A. saccharinum, and A. rubrum are fast-growing and shade-intolerant soft maples, whereas A. nigrum and A. saccharum are slow-growing and shade-tolerant hard maples. It was hypothesized that the ruderal and soft maples would have lower modulus of elasticity (MOE) and modulus of rupture (MOR), but higher maximum specific conductivity (K(s max)) than hard maples. Many anatomical and general morphological characteristics were measured in an attempt to correlate them to water transport and/or mechanical strength differences between species. No difference was found between species in vessel diameter, fiber wall thickness, initial hydraulic conductivity (K(h initial)), specific conductivity (K(s max)), native percent embolism, or Huber value. Similarly, no trade-off was found between K(s max) and MOE or MOR across the genus. However, fiber lumen diameter was inversely correlated to MOE and MOR. Surprisingly, percentage of ray parenchyma was positively related to MOE. The results suggest transport/mechanical trade-offs do not occur in Acer and differences in mechanical properties may be due to fiber lumen differences that do not influence the efficient transport of water.

The 120-yr period for Dr. Beal's seed viability experiment.

After 120 yr of burial in moist, well-aerated sand, 23 seeds of Verbascum blattaria and two seeds of a Verbascum sp. germinated and produced normal plants (50% germination for Verbascum). After a 6-wk cold treatment, a single seed of Malva rotundifolia germinated also, producing a normal plant (2% germination). Plants were grown to maturity in a greenhouse, and flowering was induced by exposure to a 6-wk cold treatment. Flowers were artificially pollinated to produce seed of both Verbascum blattaria and Malva rotundifolia. The Verbascum sp. failed to set seed. Collected seeds were subsequently germinated, producing normal plants. F(1) seeds of V. blattaria had a germination of 64%. Seeds (6%) of M. rotundifolia germinated after a cold treatment.

Thigmomorphogenesis: changes in the morphology and mechanical properties of two Populus hybrids in response to mechanical perturbation.

To identify hybrid-specific differences in developmental response to mechanical perturbation (MP), we compared the effects of stem flexure on several morphological and mechanical properties of two Populus trichocarpa Torr. & A. Gray x P. deltoides Bartr. ex Marsh. hybrids, 47-174 and 11-11. In response to the MP treatment, both hybrids exhibited a significant increase in radial growth, especially in the direction of the MP (47-174, P = 0.0001; 11-11, P = 0.002), and a significant decrease in height to diameter growth ratio (P = 0.0001 for both hybrids), suggesting that MP-treated stems are more tapered than control stems. A direct consequence of the MP-induced increase in radial growth was a significant increase in flexural rigidity (EI, N mm(2)) in stems of both hybrids (47-174, P = 0.0001; 11-11, P = 0.009). Both control and MP-treated stems of Hybrid 47-174 had significantly greater height to diameter ratios and EI values than the corresponding stems of Hybrid 11-11 (11-11 stem ratios and EI values were 85 and 76%, respectively, of those of 47-174). In Hybrid 47-174, Young's modulus of elasticity (E, N mm(-2)), a measure of stem flexibility, for MP-treated stems was only 80% of the control value (P = 0.0034), whereas MP had no significant effect on E of stems of Hybrid 11-11 (P = 0.2720). Differences in flexure response between the hybrids suggest that Hybrid 47-174 can produce a stem that is more tolerant of wind-induced flexure by altering both stem allometry and material properties, whereas Hybrid 11-11 relies solely on changes in stem allometry for enhanced stability under MP conditions.

Thigmomorphogenesis: a dose response to flexing in Ulmus americana seedlings.

Six-week-old half-sib seedlings of Ulmus americana L. were subjected to different amounts of flexure daily for 3 weeks under controlled greenhouse conditions. The daily flexure treatments were: no flexing in a staked stem, minimal flexing in a non-staked stem, and five, 10, 20, 40, and 80 flexures. Seedling height and diameter growth and average leaf area were determined before and after the treatments. The ratio of the change in height growth (DeltaH) to the change in diameter growth (DeltaD; (DeltaH:DeltaD)) before and after the 3-week treatments were calculated. At the end of the 3-week experiment, staked seedlings were significantly taller and had smaller stem diameters than all of the flexed seedlings. Height growth tended to decrease exponentially with increased flexure, with significant differences between the extremes of treatment. All of the flexure treatments significantly increased stem diameter compared to staked seedlings. The DeltaH:DeltaD ratio exhibited an exponential function in response to increased flexure. Average leaf area decreased with increased flexure, and seedlings in the 40x and 80x flexure treatments had significantly less leaf area than seedlings in all of the other treatments. These data are similar to the dose responses previously observed in herbaceous species. The finding that trees exhibit greater sensitivity to low doses of flexure than to high doses of flexure indicates that slight exposure to wind may result in a large initial alteration in stem morphology, producing a thigmomorphogenetic effect. Trees will continue to respond to increasing amounts of mechanical stress, but at an exponentially declining rate. Declining leaf areas in response to increasing amounts of mechanical stress may result in a decrease in available photosynthate, resulting in a tree of smaller stature compared to trees exposed to lower amounts of mechanical loading.