Cyclanilide Induces Lateral Bud Outgrowth by Modulating Cytokinin Biosynthesis and Signalling Pathways in Apple Identified via Transcriptome Analysis

Cyclanilide (CYC), a plant growth regulator, is a potent shoot branching agent in apple. However, its mechanism remains unclear. The current study revealed that CYC treatment resulted in massive reprogramming of the axillary bud transcriptome, implicating several hormones in the response. We observed a marked increase (approximately 2-fold) in the level of zeatin riboside and a significant decrease (approximately 2-fold) in the level of abscisic acid (ABA). Zeatin metabolism gene cytokinin (CTK) oxidase 1 (CKX 1) was down-regulated at 168 h after CYC treatment compared with the control. Weighted gene co-expression network analysis of differentially expressed genes demonstrated the turquoise module clusters exhibited the highest positive correlation with zeatin riboside (r = 0.92) and the highest negative correlation with ABA (r = −0.8). A total of 37 genes were significantly enriched in the plant hormone signal transduction pathway in the turquoise module. Among them, the expressions of CTK receptor genes WOODEN LEG and the CTK type-A response regulators genes ARR3 and ARR9 were up-regulated. ABA signal response genes protein phosphatase 2C genes ABI2 and ABI5 were down-regulated in lateral buds after CYC treatment at 168 h. In addition, exogenous application of 6-benzylaminopurine (6-BA, a synthetic type of CTK) and CYC enhanced the inducing effect of CYC, whereas exogenous application of lovastatin (a synthetic type of inhibitor of CTK biosynthesis) or ABA and CYC weakened the promoting effect of CYC. These results collectively revealed that the stimulation of bud growth by CYC might involve CTK biosynthesis and signalling, including genes CKX1 and ARR3/9, which provided a direction for further study of the branching promoting mechanism of CYC.

Expression Analysis of DgD14, DgBRC1 and DgLsL in the Process of Chrysanthemum Lateral Bud Formation

The growth of lateral bud can greatly affect the development of apical bud and reduce the quality of single-flower cut chrysanthemum. However, the wide use of artificial bud removal in production leads to the increase on production cost. Therefore, it is important to study the lateral bud development mechanism in chrysanthemum for plant type regulation and genetic improvement. Auxin (IAA), cytokinins (CKs) and strigolactones (SLs) have direct or indirect effects on the formation of lateral buds. D14, BRC1 and LsL are key factors regulating the signal pathways of hormones, but their regulation mechanisms on the development of lateral buds in chrysanthemum are still unclear. In this study, single-flower cut chrysanthemum ‘Jinba’ and spray cut chrysanthemum ‘Fenyan’ were used as experimental materials. Quantitative real-time PCR was used to observe the effects of apical bud removal and exogenous hormones on the growth of lateral buds and the expression levels of DgD14, DgBRC1 and DgLsL, so as to clarify the expression characteristics of three genes in the process of lateral bud formation. The results showed that GA was effective in promoting the growth of lateral buds, whereas IAA and ABA had little effects on lateral bud growth or even inhibited. Removing apical dominance can significantly affect the expression levels of three genes, which regulated the formation and elongation of lateral buds. Additionally, the three genes showed different responses to different hormone treatments. DgD14 had a significant response to GA, but a gentle response to ABA. The expression levels of DgBRC1 varied in different trends, and it responded to IAA in a more dramatic way. The levels of DgLsL reached the peaks quickly before decreased in most experimental groups, and its response to GA was extraordinary severe.

Septin Assembly and Remodeling at the Cell Division Site During the Cell Cycle

The septin family of proteins can assemble into filaments that further organize into different higher order structures to perform a variety of different functions in different cell types and organisms. In the budding yeast Saccharomyces cerevisiae, the septins localize to the presumptive bud site as a cortical ring prior to bud emergence, expand into an hourglass at the bud neck (cell division site) during bud growth, and finally “split” into a double ring sandwiching the cell division machinery during cytokinesis. While much work has been done to understand the functions and molecular makeups of these structures, the mechanisms underlying the transitions from one structure to another have largely remained elusive. Recent studies involving advanced imaging and in vitro reconstitution have begun to reveal the vast complexity involved in the regulation of these structural transitions, which defines the focus of discussion in this mini-review.

A preferred sequence for organelle inheritance during polarized cell growth

Some organelles cannot be synthesized anew, so they are segregated into daughter cells during cell division. In Saccharomyces cerevisiae, daughter cells bud from mother cells and are populated by organelles inherited from the mothers. To determine whether this organelle inheritance occurs in a stereotyped manner, we tracked organelles using fluorescence microscopy. We describe a program for organelle inheritance in budding yeast. The cortical endoplasmic reticulum (ER) and peroxisomes are inherited concomitant with bud emergence. Next, vacuoles are inherited in small buds, followed closely by mitochondria. Finally, the nucleus and perinuclear ER are inherited when buds have nearly reached their maximal size. Because organelle inheritance timing correlates with bud morphology, which is coupled to the cell cycle, we tested whether disrupting the cell cycle alters organelle inheritance order. By arresting cell cycle progression but allowing continued bud growth, we determined that organelle inheritance still occurs when DNA replication is blocked, and that the general inheritance order is maintained. Thus, organelle inheritance follows a preferred order during polarized cell division and does not require completion of S-phase.

Decapitation Experiments Combined with the Transcriptome Analysis Reveal the Mechanism of High Temperature on Chrysanthemum Axillary Bud Formation

Temperature is an important factor that largely affects the patterns of shoot branching in plants. However, the effect and mechanism of temperature on axillary bud development in chrysanthemum remains poorly defined. The purpose of the present study is to investigate the effect of high temperature on the axillary bud growth and the mechanism of axillary bud formation in chrysanthemum. Decapitation experiments combined with the transcriptome analysis were designed. Results showed that the axillary bud length was significantly inhibited by high temperature. Decapitation of primary shoot (primary decapitation) resulted in slower growth of axillary buds (secondary buds) under 35 °C. However, secondary decapitation resulted in complete arrest of tertiary buds at high temperature. These results demonstrated that high temperature not only inhibited axillary bud formation but also retarded bud outgrowth in chrysanthemum. Comparative transcriptome suggested differentially expressed gene sets and identified important modules associated with bud formation. This research helped to elucidate the regulatory mechanism of high temperature on axillary bud growth, especially bud formation in chrysanthemum. Meanwhile, in-depth studies of this imperative temperature signaling can offer the likelihood of vital future applications in chrysanthemum breeding and branching control.

Control of axillary bud growth in tobacco through toxin gene expression system

The control of axillary bud development after removing the terminal buds (topping) of plants is a research hotspot, and the control of gene expression, like switching on and off, allows us to further study biological traits of interest, such as plant branching and fertility. In this study, a toxin gene control system for plants based on dexamethasone (DEX) induction was constructed, and the positive transgenic tobacco exhibited growth retardation in the application area (axillary bud). The expression level of the lethal Diphtheria toxin A (DTA) gene under different DEX concentrations at different application days was analyzed. The highest expression levels appeared at 5 days after the leaf injection of DEX. The DTA transcripts were induced by 5 µM DEX and peaked in response to 50 µM DEX at 5 days after leaf injection. Here, a chemical induction system, combined with a toxin gene, were used to successfully control the growth of tobacco axillary buds after topping. The DTA expression system under DEX induction was sensitive and efficient, therefore, can be used to control axillary bud growth and development in tobacco.

Developmental influence on evolutionary rates and the origin of placental mammal tooth complexity

Development has often been viewed as a constraining force on morphological adaptation, but its precise influence, especially on evolutionary rates, is poorly understood. Placental mammals provide a classic example of adaptive radiation, but the debate around rate and drivers of early placental evolution remains contentious. A hallmark of early dental evolution in many placental lineages was a transition from a triangular upper molar to a more complex upper molar with a rectangular cusp pattern better specialized for crushing. To examine how development influenced this transition, we simulated dental evolution on “landscapes” built from different parameters of a computational model of tooth morphogenesis. Among the parameters examined, we find that increases in the number of enamel knots, the developmental precursors of the tooth cusps, were primarily influenced by increased self-regulation of the molecular activator (activation), whereas the pattern of knots resulted from changes in both activation and biases in tooth bud growth. In simulations, increased activation facilitated accelerated evolutionary increases in knot number, creating a lateral knot arrangement that evolved at least ten times on placental upper molars. Relatively small increases in activation, superimposed on an ancestral tritubercular molar growth pattern, could recreate key changes leading to a rectangular upper molar cusp pattern. Tinkering with tooth bud geometry varied the way cusps initiated along the posterolingual molar margin, suggesting that small spatial variations in ancestral molar growth may have influenced how placental lineages acquired a hypocone cusp. We suggest that development could have enabled relatively fast higher-level divergence of the placental molar dentition.

Cell influx and contractile actomyosin force drive mammary bud growth and invagination

The mammary gland develops from the surface ectoderm during embryogenesis and proceeds through morphological phases defined as placode, hillock, bud, and bulb stages followed by branching morphogenesis. During this early morphogenesis, the mammary bud undergoes an invagination process where the thickened bud initially protrudes above the surface epithelium and then transforms to a bulb and sinks into the underlying mesenchyme. The signaling pathways regulating the early morphogenetic steps have been identified to some extent, but the underlying cellular mechanisms remain ill defined. Here, we use 3D and 4D confocal microscopy to show that the early growth of the mammary rudiment is accomplished by migration-driven cell influx, with minor contributions of cell hypertrophy and proliferation. We delineate a hitherto undescribed invagination mechanism driven by thin, elongated keratinocytes—ring cells—that form a contractile rim around the mammary bud and likely exert force via the actomyosin network. Furthermore, we show that conditional deletion of nonmuscle myosin IIA (NMIIA) impairs invagination, resulting in abnormal mammary bud shape.

Regulation of 2,4-D Isooctyl Ester on Triticum aestivum and Aegilops tauschii Tillering and Endogenous Phytohormonal Responses

Tillering is an important agronomic trait essential for the yield of Triticum aestivum and the propagation of Aegilops tauschii. However, the effect of phytohormones on T. aestivum and Ae. tauschii tillering and the underlying regulatory mechanisms remain poorly understood. In the study, we found that T. aestivum and Ae. tauschii exhibited different tillering sensitivities to the auxin herbicide 2,4-D isooctyl ester. At 3 days post-application, tiller bud growth was inhibited by 77.50% in T. aestivum, corresponding to 2.0-fold greater inhibition than that in Ae. tauschii (38.71%). Transcriptome analysis showed that differentially expressed genes (DEGs) in the T. aestivum response to 2,4-D isooctyl ester were mainly enriched in plant hormone metabolism and signal transduction pathways, but similar changes were not observed in Ae. tauschii. Among that, the auxin biosynthesis and signaling induced by 2,4-D isooctyl ester was quite different between the two species. A total of nine candidate genes involved in varied tillering responses were selected from the DEGs and validated by quantitative real-time PCR. Endogenous hormone levels were assayed to further verify the RNA-seq results. After 2,4-D isooctyl ester treatment, a significant increase in abscisic acid (ABA) levels was observed in T. aestivum, whereas ABA levels were relatively stable in Ae. tauschii. The herbicide induced more cytokinin (CTK) accumulation in Ae. tauschii than in T. aestivum. External ABA clearly restricted tiller bud growth in both T. aestivum and Ae. tauschii, while 6-benzyl aminopurine had no significant effect. These results indicate that ABA and CTK may be related with 2,4-D isooctyl ester-regulated tillering differences between the two species, which will help to further understand the mechanism of the auxin-mediated regulation of tillering