Trichome Gene Research Articles

TrichomeYOLO: A Neural Network for Automatic Maize Trichome Counting.

Plant trichomes are epidermal structures with a wide variety of functions in plant development and stress responses. Although the functional importance of trichomes has been realized, the tedious and time-consuming manual phenotyping process greatly limits the research progress of trichome gene cloning. Currently, there are no fully automated methods for identifying maize trichomes. We introduce TrichomeYOLO, an automated trichome counting and measuring method that uses a deep convolutional neural network, to identify the density and length of maize trichomes from scanning electron microscopy images. Our network achieved 92.1% identification accuracy on scanning electron microscopy micrographs of maize leaves, which is much better performed than the other 5 currently mainstream object detection models, Faster R-CNN, YOLOv3, YOLOv5, DETR, and Cascade R-CNN. We applied TrichomeYOLO to investigate trichome variations in a natural population of maize and achieved robust trichome identification. Our method and the pretrained model are open access in Github (https://github.com/yaober/trichomecounter). We believe TrichomeYOLO will help make efficient trichome identification and help facilitate researches on maize trichomes.

Trichomes on female reproductive tract: rapid diversification and underlying gene regulatory network in Drosophila suzukii and its related species

BackgroundThe ovipositors of some insects are external female genitalia, which have their primary function to deliver eggs. Drosophila suzukii and its sibling species D. subpulchrella are known to have acquired highly sclerotized and enlarged ovipositors upon their shifts in oviposition sites from rotting to ripening fruits. Inside the ovipositor plates, there are scale-like polarized protrusions termed “oviprovector scales” that are likely to aid the mechanical movement of the eggs. The size and spatial distribution of the scales need to be rearranged following the divergence of the ovipositors. In this study, we examined the features of the oviprovector scales in D. suzukii and its closely related species. We also investigated whether the scales are single-cell protrusions comprised of F-actin under the same conserved gene regulatory network as the well-characterized trichomes on the larval cuticular surface.ResultsThe oviprovector scales of D. suzukii and D. subpulchrella were distinct in size and spatial arrangement compared to those of D. biarmipes and other closely related species. The scale numbers also varied greatly among these species. The comparisons of the size of the scales suggested a possibility that the apical cell area of the oviprovector has expanded upon the elongation of the ovipositor plates in these species. Our transcriptome analysis revealed that 43 out of the 46 genes known to be involved in the trichome gene regulatory network are expressed in the developing female genitalia of D. suzukii and D. subpulchrella. The presence of Shavenbaby (Svb) or svb was detected in the inner cavity of the developing ovipositors of D. melanogaster, D. suzukii, and D. subpulchrella. Also, shavenoid (sha) was expressed in the corresponding patterns in the developing ovipositors and showed differential expression levels between D. suzukii and D. subpulchrella at 48 h APF.ConclusionsThe oviprovector scales have divergent size and spatial arrangements among species. Therefore, these scales may represent a rapidly diversifying morphological trait of the female reproductive tract reflecting ecological contexts. Furthermore, our results showed that the gene regulatory network underlying trichome formation is also utilized to develop the rapidly evolving trichomes on the oviprovectors of these flies.

Methyl jasmonate treatment, aphid resistance assay, and transcriptomic analysis revealed different herbivore defensive roles between tobacco glandular and non-glandular trichomes.

Methyl jasmonate treatment and aphid resistance assays reveal different roles in herbivore defensive responses between tobacco glandular and non-glandular trichomes. These roles correlate with trichome gene expression patterns. In plants, trichomes greatly contribute to biotic stress resistance. To better understand the different defensive functions between glandular and non-glandular trichomes, we used Nicotiana tabacum as a model. This species bears three types of trichomes: long and short stalk glandular trichomes (LGT and SGT, respectively), and non-glandular trichomes (NGT). Tobacco accession T.I.1068 (lacking NGT) and T.I.1112 (lacking LGT) were used for the experiment. After methyl jasmonate (MeJA) treatment, LGT formation was promoted not only in T.I.1068, but also in T.I.1112, whereas NGT remained absent in T.I.1068, and was slightly reduced in T.I.1112. Diterpenoids, which play important roles in herbivore resistance, accumulated abundantly in T.I.1068 and were elevated by MeJA; however, they were not found in T.I.1112 but became detectable after MeJA treatment. The aphid resistance of T.I.1068 was higher than that of T.I.1112, and both were enhanced by MeJA, which was closely correlated with LGT density. Trichomes detached from T.I.1068 and T.I.1112 were used for RNA-Seq analysis, the results showed that pentose phosphate, photosynthesis, and diterpenoid biosynthesis genes were much more expressed in T.I.1068 than in T.I.1112, which was consistent with the vigorous diterpenoid biosynthesis in T.I.1068. In T.I.1112, citrate cycle, propanoate, and glyoxylate metabolism processes were enriched, and some defensive protein genes were expressed at higher levels than those in T.I.1068.These results suggested that LGT plays a predominant role in aphid resistance, whereas NGT could strengthen herbivore resistance by accumulating defensive proteins, and the roles of LGT and NGT are associated with their gene expression patterns.

Gene expression underlying floral epidermal specialization in Aristolochia fimbriata (Aristolochiaceae).

The epidermis constitutes the outermost tissue of the plant body. Although it plays major structural, physiological and ecological roles in embryophytes, the molecular mechanisms controlling epidermal cell fate, differentiation and trichome development have been scarcely studied across angiosperms, and remain almost unexplored in floral organs. In this study, we assess the spatio-temporal expression patterns of GL2, GL3, TTG1, TRY, MYB5, MYB6, HDG2, MYB106-like, WIN1 and RAV1-like homologues in the magnoliid Aristolochia fimbriata (Aristolochiaceae) by using comparative RNA-sequencing and in situ hybridization assays. Genes involved in Aristolochia fimbriata trichome development vary depending on the organ where they are formed. Stem, leaf and pedicel trichomes recruit most of the transcription factors (TFs) described above. Conversely, floral trichomes only use a small subset of genes including AfimGL2, AfimRAV1-like, AfimWIN1, AfimMYB106-like and AfimHDG2. The remaining TFs, AfimTTG1, AfimGL3, AfimTRY, AfimMYB5 and AfimMYB6, are restricted to the abaxial (outer) and the adaxial (inner) pavement epidermal cells. We re-evaluate the core genetic network shaping trichome fate in flowers of an early-divergent angiosperm lineage and show a morphologically diverse output with a simpler genetic mechanism in place when compared to the models Arabidopsis thaliana and Cucumis sativus. In turn, our results strongly suggest that the canonical trichome gene expression appears to be more conserved in vegetative than in floral tissues across angiosperms.

Molecular mechanism and physiological function of trichome formation in plant

Trichome, as a protective barrier against natural hazards, plays an indispensable role in resisting ultraviolet radiation, pathogen invasion, herbivores feeding and water excessive transpiration during the process of plant growth and development. Trichomes may be unicellular or multicellular and some plants such as Arabidopsis thaliana may also have branches, according to the presence or absence of glands, it can be divided into glandular or non-glandular, the former can accumulate and secrete some alkaloids, such as nicotine and terpenoids, which have an exclusion effect on insects, while the latter can be enhanced the stress resistance during abiotic stress, such as extreme high and low temperatures, ultraviolet radiation, thereby promoting and controlling the normal growth of plants. The current study shows that the formation of trichome is co-regulation by many types and multiple genes, thus form a complex regulation network. The mechanism of the regulation and genes mining of the trichome in the dicotyledonous has made substantial progress in recent years, especially in Arabidopsis , Trichome formation in Athaliana is thought to be regulated by a competitive system, including the promotion and inhibition of the gene activities. More and more studies have shown that there are a variety of different genes can regulate the trichome formation in different plants. The genes controling trichome formation are generally a class of MYB transcription factors in Arabidopsis [31] , whereas a HD-Zip protein, Woolly (Wo) that interacts with Cyclin B2, plays an essential role for trichome formation and embryonic development in tomato. In addition, the Glabrous Rice 1(GLR1), which encodes a WUS-like homeobox gene (WOX), also regulates the formation of trichomes in rice. It was proposed that Trichome can also interact with the environment under the development process of plants, such as light refraction, light quantum radiation interception, leaf heat balance, wettability, droplet retention, water uptake and gas exchange. Leaf trichome density is variable and determined by genetic and environmental factors, climate drivers such as soil water deficit and high temperature can increase the trichome density. The studies have shown that leaf trichomes can reflect broad-spectrum electromagnetic radiation, and reducing light absorptance and modulating energy balance. Except as described above, trichome can promote gas exchange and limit pathogen growth and spread on leaves. According to the physiological characteristics of the trichome, the crop cultivation, production and utilization have important economic value and significance. In this article, We systematically reviewed and analyzed a series of related studies on the progress and physiological characteristics of genes which related to the regulation of plant trichome formation in recent years, The aim of this review was to elucidate the regulation mechanism of the trichome formation and the physiological and biochemical characteristics, so as to provide an important theoretical basis for further research on the excavation and functional characteristics of the trichome genes.

Genome-Wide Survey Reveals Transcriptional Differences Underlying the Contrasting Trichome Phenotypes of Two Sister Desert Poplars.

Trichomes, which are widely used as an important diagnostic characteristic in plant species delimitation, play important roles in plant defense and adaptation to adverse environments. In this study, we used two sister poplar species, Populus pruinosa and Populus euphratica—which have, respectively, dense and sparse trichomes—to examine the genetic differences associated with these contrasting phenotypes. The results showed that 42 and 45 genes could be identified as candidate genes related to trichomes in P. pruinosa and P. euphratica, respectively; most of these genes possessed high degrees of diversification in their coding sequences, but they were similar in intron/exon structure in the two species. We also found that most of the candidate trichome genes were expressed at higher levels in P. pruinosa, which has dense trichomes, than in P. euphratica, where there are few trichomes. Based on analyses of transcriptional profiles, a total of 195 genes, including many transcription factors, were found to show distinct differences in expression. The results of gene function annotation suggested that the genes identified as having contrasting levels of expression level are mainly associated with trichome elongation, ATPase activity, and hormone transduction. Changes in the expression of these and other related genes with high sequence diversification may have contributed to the contrast in the pattern of trichome phenotypes between the two species.

Hairy Canola (Brasssica napus) re-visited: Down-regulating TTG1 in an AtGL3-enhanced hairy leaf background improves growth, leaf trichome coverage, and metabolite gene expression diversity.

BackgroundThrough evolution, some plants have developed natural resistance to insects by having hairs (trichomes) on leaves and other tissues. The hairy trait has been neglected in Brassica breeding programs, which mainly focus on disease resistance, yield, and overall crop productivity. In Arabidopsis, a network of three classes of proteins consisting of TTG1 (a WD40 repeat protein), GL3 (a bHLH factor) and GL1 (a MYB transcription factor), activates trichome initiation and patterning. Introduction of a trichome regulatory gene AtGL3 from Arabidopsis into semi-glabrous Brassica napus resulted in hairy canola plants which showed tolerance to flea beetles and diamondback moths; however plant growth was negatively affected. In addition, the role of BnTTG1 transcription in the new germplasm was not understood.ResultsHere, we show that two ultra-hairy lines (K-5-8 and K-6-3) with BnTTG1 knock-down in the hairy AtGL3+ B. napus background showed stable enhancement of trichome coverage, density, and length and restored wild type growth similar to growth of the semi-glabrous Westar plant. In contrast, over-expression of BnTTG1 in the hairy AtGL3+ B. napus background gave consistently glabrous plants of very low fertility and poor stability, with only one glabrous plant (O-3-7) surviving to the T3 generation. Q-PCR trichome gene expression data in leaf samples combining several leaf stages for these lines suggested that BnGL2 controlled B. napus trichome length and out-growth and that strong BnTTG1 transcription together with strong GL3 expression inhibited this process. Weak expression of BnTRY in both glabrous and trichome-bearing leaves of B. napus in the latter Q-PCR experiment suggested that TRY may have functions other than as an inhibitor of trichome initiation in the Brassicas. A role for BnTTG1 in the lateral inhibition of trichome formation in neighbouring cells was also proposed for B. napus. RNA sequencing of first leaves identified a much larger array of genes with altered expression patterns in the K-5-8 line compared to the hairy AtGL3+B. napus background (relative to the Westar control plant). These genes particularly included transcription factors, protein degradation and modification genes, but also included pathways that coded for anthocyanins, flavonols, terpenes, glucosinolates, alkaloids, shikimates, cell wall biosynthesis, and hormones. A 2nd Q-PCR experiment was conducted on redox, cell wall carbohydrate, lignin, and trichome genes using young first leaves, including T4 O-3-7-5 plants that had partially reverted to yield two linked growth and trichome phenotypes. Most of the trichome genes tested showed to be consistant with leaf trichome phenotypes and with RNA sequencing data in three of the lines. Two redox genes showed highest overall expression in K-5-8 leaves and lowest in O-3-7-5 leaves, while one redox gene and three cell wall genes were consistently higher in the two less robust lines compared with the two robust lines.ConclusionThe data support the strong impact of BnTTG1 knockdown (in the presence of strong AtGL3 expression) at restoring growth, enhancing trichome coverage and length, and enhancing expression and diversity of growth, metabolic, and anti-oxidant genes important for stress tolerance and plant health in B. napus. Our data also suggests that the combination of strong (up-regulated) BnTTG1 expression in concert with strong AtGL3 expression is unstable and lethal to the plant.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-015-0680-5) contains supplementary material, which is available to authorized users.

A landscape of hairy and twisted: hunting for new trichome mutants in the Saskatoon Arabidopsis T-DNA population.

A total of 88 new Arabidopsis lines with trichome variation were recovered by screening 49,200 single-seed descent T3 lines from the SK activation-tagged population and from a new 20,000-line T-DNA insertion population (called pAG). Trichome variant lines were classified into 12 distinct phenotype categories. Single or multiple T-DNA insertion sites were identified for 89% of these mutant lines. Alleles of the well-known trichome genes TRY, GL2 and TTG1 were recovered with atypical phenotype variation not reported previously. Moreover, atypical gene expression profiles were documented for two additional mutants specifying TRY and GL2 disruptions. In remaining mutants, ten lines were disrupted in genes coding for proteins not implicated in trichome development, five were disrupted in hypothetical proteins and 11 were disrupted in proteins with unknown function. The collection represents new opportunities for the plant biology community to define trichome development more precisely and to refine the function of individual trichome genes.

Brassica villosa, a system for studying non-glandular trichomes and genes in the Brassicas.

Brassica villosa is a wild Brassica C genome species with very dense trichome coverage and strong resistance to many insect pests of Brassica oilseeds and vegetables. Transcriptome analysis of hairy B. villosa leaves indicated higher expression of several important trichome initiation genes compared with glabrous B. napus leaves and consistent with the Arabidopsis model of trichome development. However, transcripts of the TRY inhibitory gene in hairy B. villosa were surprisingly high relative to B. napus and relative transcript levels of SAD2, EGL3, and several XIX genes were low, suggesting potential ancillary or less important trichome-related roles for these genes in Brassica species compared with Arabidopsis. Several antioxidant, calcium, non-calcium metal and secondary metabolite genes also showed differential expression between these two species. These coincided with accumulation of two alkaloid-like compounds, high levels of calcium, and other metals in B. villosa trichomes that are correlated with the known tolerance of B. villosa to high salt and the calcium-rich natural habitat of this wild species. This first time report on the isolation of large amounts of pure B. villosa trichomes, on trichome content, and on relative gene expression differences in an exceptionally hairy Brassica species compared with a glabrous species opens doors for the scientific community to understand trichome gene function in the Brassicas and highlights the potential of B. villosa as a trichome research platform.

Converting restriction fragment length polymorphism to single‐strand conformation polymorphism markers and its application in the fine mapping of a trichome gene in cotton

AbstractA well‐characterized and systematically organized collection of genetic markers is crucial in the study of any crop species. It is the basis of map‐based gene cloning and crop improvements through marker‐assisted selections. Single‐strand conformation polymorphism (SSCP) has been a robust way of discovering new polymorphisms in marker development without the requirement of sequencing. Here, we report the first approach of applying SSCP marker discovery methods in the genetic map construction and gene mapping of cotton species. A total of 80 restriction fragment length polymorphism (RFLP) markers were selected from a region on published cotton genetic maps around the T1 gene related to cotton trichome. Among the 80 RFLPs, 28 showed polymorphisms through SSCP, showing a polymorphic rate of approximately 35%, which is much higher than that of simple sequence repeat (SSR) markers in the same region (7.8%). By integrating these newly generated SSCP markers, a detailed genetic map was reconstructed around this region using an F2 population derived from a cross between Gossypium arboreum and G. herboceum. The reconstructed region comprises 22 SSCP markers, eight SSR markers and the T1 gene, spanning 21.6 cM. The marker order of the new map agrees well with published reference RFLP maps. The above results suggest that SSCP method can be applied very efficiently and reliably to the marker development of cotton genomes. It will prove to be even more valuable and robust after the public release of cotton whole‐genome sequences.

Coexistence of trichome variation in a natural plant population: a combined study using ecological and candidate gene approaches.

The coexistence of distinct phenotypes within populations has long been investigated in evolutionary ecology. Recent studies have identified the genetic basis of distinct phenotypes, but it is poorly understood how the variation in candidate loci is maintained in natural environments. In this study, we examined fitness consequences and genetic basis of variation in trichome production in a natural population of Arabidopsis halleri subsp. gemmifera. Half of the individuals in the study population produced trichomes while the other half were glabrous, and the leaf beetle Phaedon brassicae imposed intensive damage to both phenotypes. The fitness of hairy and glabrous plants showed no significant differences in the field during two years. A similar result was obtained when sibling hairy and glabrous plants were transplanted at the same field site, whereas a fitness cost of trichome production was detected under a weak herbivory condition. Thus, equivalent fitness of hairy and glabrous plants under natural herbivory allows their coexistence in the contemporary population. The pattern of polymorphism of the candidate trichome gene GLABROUS1 (GL1) showed no evidence of long-term maintenance of trichome variation within the population. Although balancing selection under fluctuating biotic environments is often proposed to explain the maintenance of defense variation, the lack of clear evidence of balancing selection in the study population suggests that other factors such as gene flow and neutral process may have played relatively large roles in shaping trichome variation at least for the single population level.

Gene expression profile analysis of tobacco leaf trichomes

BackgroundLeaf trichomes of Nicotiana tabacum are distinguished by their large size, high density, and superior secretion ability. They contribute to plant defense response against biotic and abiotic stress, and also influence leaf aroma and smoke flavor. However, there is limited genomic information about trichomes of this non-model plant species.ResultsWe have characterized Nicotiana tabacum leaf trichome gene expression using two approaches. In the first, a trichome cDNA library was randomly sequenced, and 2831 unique genes were obtained. The most highly abundant transcript was ribulose bisphosphate carboxylase (RuBisCO). Among the related sequences, most encoded enzymes involved in primary metabolism. Secondary metabolism related genes, such as isoprenoid and flavonoid biosynthesis-related, were also identified. In the second approach, a cDNA microarray prepared from these 2831 clones was used to compare gene expression levels in trichome and leaf. There were 438 differentially expressed genes between trichome and leaves-minus-trichomes. Of these, 207 highly expressed genes in tobacco trichomes were enriched in second metabolic processes, defense responses, and the metabolism regulation categories. The expression of selected unigenes was confirmed by semi-quantitative RT-PCR analysis, some of which were specifically expressed in trichomes.ConclusionThe expression feature of leaf trichomes in Nicotiana tabacum indicates their metabolic activity and potential importance in stress resistance. Sequences predominantly expressed in trichomes will facilitate gene-mining and metabolism control of plant trichome.

Genetic interaction between glabra3‐shapeshifter and siamese in Arabidopsis thaliana converts trichome precursors into cells with meristematic activity

The identity of many genes required for trichome differentiation is known. This paper describes a novel interaction between mutant alleles of two such genes. One of the alleles, called gl3-sst, is derived from the GL3 locus, which encodes a basic helix-loop-helix type transcription factor. The mutation in the gl3-sst protein modifies its ability to form a complex with the GL1 protein (a MYB transcription factor required for trichome formation), leading to changes in gene expression compared with wild type during gl3-sst mutant trichome development. The other mutant allele, sim, is a likely loss of function allele derived from the SIM locus, which is predicted to encode a negative regulator of D-type cyclin activity. The gl3-sst sim double mutant exhibits mounds of cells derived from the proliferation of single trichome precursors. The ectopic expression of a D-type cyclin gene in gl3-sst mimics the double mutant phenotype. Thus, an interaction between altered trichome gene expression caused by the gl3-sst mutation and relaxed regulation of D-type cyclin activity in the double mutant converted a non-dividing cell into a novel highly proliferating cell type.

A versatile promoter for the expression of proteins in glandular and non-glandular trichomes from a variety of plants

A DNA regulatory fragment was isolated from the promoter region of the OASA1 gene, encoding the cytosolic O-acetylserine(thiol)lyase enzyme that is highly expressed in Arabidopsis thaliana trichomes. This DNA fragment has been named an ATP fragment and comprises 1435 bp of the genomic region upstream of the OASA1 gene and 375 bp of the transcriptional initiation start site containing the first intron of the gene. The ATP fragment, fused to the green fluorescent protein (GFP) and beta-glucuronidase (GUS) reporter genes, is able to drive high-level gene expression in A. thaliana trichomes. Deletion analysis of the ATP fragment determined that the region from -266 to -66 contains regulatory elements required for trichome expression. In addition, the region from +112 to +375, comprising the first intronic region of the gene, is also essential for trichome gene expression. Expression of the full-length ATP fragment in tobacco and peppermint shows that this fragment is also able to drive expression in glandular trichomes and suggests additional biotechnological applications for this promoter.

Transcriptome analysis of alfalfa glandular trichomes

Glandular trichomes are a major site of plant natural product synthesis and accumulation for protection against insect predation. However, to date few studies have attempted to obtain a global view of trichome gene expression. Two contrasting approaches have been adopted to investigate genes expressed in glandular trichomes from alfalfa (Medicago sativa L.). In the first approach, 5,674 clones from an alfalfa glandular trichome cDNA library were sequenced. The most highly abundant expressed sequence tag (EST) corresponded to a lipid transfer protein. The presence of ESTs corresponding to enzymes for all steps in the biosynthesis of flavonoids suggests that these are important metabolites in alfalfa trichome biology, as confirmed by histochemistry and metabolite profiling. No ESTs corresponded to enzymes of cyclized terpenoid biosynthesis. In a second approach, microarray analysis was used to compare levels of alfalfa transcripts corresponding to 16,086 Medicago truncatula A17 genes in stems with and without trichomes. This revealed over 1,000 genes with strong preferential expression in the trichome fraction of the stem, 70% of which are of unknown function. These define a class of genes that are not trichome-specific, since M. truncatula A17 does not itself have glandular trichomes, but has potential importance for trichome function within the stem.

Tissue patterning of Arabidopsis cotyledons.

• Trichome and stomatal patterning are not independent events because trichomes form before stomata. We thought trichome genes might provide spatial referents to ensure proper distribution of stomata for gas exchange, and therefore studied mutants of GL1 and TRY using stomatal pattern of the entire cotyledon surface as the indicator. • Mature cotyledons were imaged by SEM, stomatal maps were generated, and data were spatially analysed. Expression of GL1 and TRY was determined in wild type and mutant samples by reverse transcriptase-PCR (RT-PCR) analysis. • At the tissue level, findings showed wild type cotyledons had a random stomatal pattern, whereas gl1-1 and try240 cotyledons had ordered and clustered stomatal patterns, respectively. Regardless of overall pattern type, c. 10% of the stomatal population - those closest to one another - were always ordered, the result of genes regulating cellular differentiation. • These results indicate epidermal cells respond to GL1 and TRY signals that affect distribution of both stomata and trichomes in postembryogenic events. The GL1 and TRY genes play dual roles in the epidermis, one role regulating epidermal tissue patterning and a second role connected with trichome development.