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.

Reduction of Early Fruit Abscission by Main-branch-girdling in Macadamia Is Related to the Favorable Status of Carbohydrates and Endogenous Hormones

Fruit abscission occurring severely in the early fruit development affects macadamia yield. Developing effective methods to improve fruit retention is a priority for macadamia cultivation and production. Girdling is an important horticultural practice that has been widely used to increase fruit yield. Previous studies have shown that girdling fails to increase macadamia yield despite enhancing the early fruit set, but few have examined the effect of girdling on its related physiological mechanism. The objective of this study was to investigate the effects of main-branch girdling (MBG) on early fruit retention and also on the levels of carbohydrates and endogenous hormones in the leaves, bearing shoots and fruit of macadamia. Herein, MBG was performed at fruit set using a single-blade knife on 9-year-old macadamia trees (Macadamia integrifolia). Results showed that MBG significantly reduced young fruit drop, concurrent with significant increases in the contents of starch in both the leaves and the bearing shoots and in glucose, fructose, and sucrose levels in the husk and seed. It was suggested that the availability of carbohydrate for fruit retention was improved by MBG. Additionally, MBG increased indole-3-acetic acid (IAA), gibberellin (GA3), and zeatin-riboside (ZR, a type of cytokinin) concentrations and decreased abscisic acid (ABA) contents in the husk and the seed, indicating that MBG reduced the early fruit drop by modifying the balance of endogenous hormones. Therefore, a positive interplay between carbohydrates and endogenous hormones induced by MBG was involved in the reduction of early fruit abscission in macadamia.

Sexual and Apogamous Species of Woodferns Show Different Protein and Phytohormone Profiles

The gametophyte of ferns reproduces either by sexual or asexual means. In the latter, apogamy represents a peculiar case of apomixis, in which an embryo is formed from somatic cells. A proteomic and physiological approach was applied to the apogamous fern Dryopteris affinis ssp. affinis and its sexual relative D. oreades. The proteomic analysis compared apogamous vs. female gametophytes, whereas the phytohormone study included, in addition to females, three apogamous stages (filamentous, spatulate, and cordate). The proteomic profiles revealed a total of 879 proteins and, after annotation, different regulation was found in 206 proteins of D. affinis and 166 of its sexual counterpart. The proteins upregulated in D. affinis are mostly associated to protein metabolism (including folding, transport, and proteolysis), ribosome biogenesis, gene expression and translation, while in the sexual counterpart, they account largely for starch and sucrose metabolism, generation of energy and photosynthesis. Likewise, ultra-performance liquid chromatography-tandem spectrometry (UHPLC-MS/MS) was used to assess the levels of indol-3-acetic acid (IAA); the cytokinins: 6-benzylaminopurine (BA), trans-Zeatine (Z), trans-Zeatin riboside (ZR), dyhidrozeatine (DHZ), dyhidrozeatin riboside (DHZR), isopentenyl adenine (iP), isopentenyl adenosine (iPR), abscisic acid (ABA), the gibberellins GA3 and GA4, salicylic acid (SA), and the brassinosteroids: brassinolide (BL) and castasterone (CS). IAA, the cytokinins Z, ZR, iPR, the gibberellin GA4, the brassinosteoids castasterone, and ABA accumulated more in the sexual gametophyte than in the apogamous one. When comparing the three apogamous stages, BA and SA peaked in filamentous, GA3 and BL in spatulate and DHRZ in cordate gametophytes. The results point to the existence of large metabolic differences between apogamous and sexual gametophytes, and invite to consider the fern gametophyte as a good experimental system to deepen our understanding of plant reproduction.

Comparison of Auxin and Cytokinins Concentrations, and the Structure of Bacterial Community between Host Twigs and Lithosaphonecrus arcoverticus Galls

Insect galls are the abnormal growth of plant tissues induced by a wide variety of galling insects and characterized by high concentrations of auxins and cytokinins. It remains unclear whether the auxins and cytokinins affect the bacterial community structure of insect galls. We determined the concentrations of indoleacetic acid (IAA) as an example of auxin, trans-zeatin riboside (tZR) and isopentenyladenine (iP) as cytokinins in Lithosaphonecrus arcoverticus (Hymenoptera: Cynipidae) galls and the galled twigs of Lithocarpus glaber (Fagaceae) using liquid chromatography–tandem mass spectrometry. Moreover, for the first time, we compared the bacterial community structure of L. arcoverticus galls and galled twigs by high-throughput sequencing, and calculated the Spearman correlation and associated degree of significance between the IAA, tZR and iP concentrations and the bacterial community structure. Our results indicated the concentrations of IAA, tZR and iP were higher in L. arcoverticus galls than in galled twigs, and positively correlated with the bacterial community structure of L. arcoverticus galls. We suggest the high concentrations of IAA, tZR and iP may affect the bacterial community structure of L. arcoverticus galls.

Exogenous Abscisic Acid Mediates Berry Quality Improvement by Altered Endogenous Plant Hormones Level in “Ruiduhongyu” Grapevine

Abscisic acid (ABA) plays a key role in fruit development and ripening in non-climacteric fruit. A variety of metabolites such as sugars, anthocyanins, fatty acids, and several antioxidants, which are regulated by various phytohormones, are important components of fruit quality in grape. Here, grape cultivar “Ruiduhongyu” was used to investigate the relationship between endogenous phytohormones and metabolites associated to grape berry quality under exogenous ABA treatment. 500 mg/L ABA significantly improved the appearance parameters and the content of many metabolites including sugar, anthocyanin, and other compounds. Exogenous ABA also increased the contents of ABA, auxin (IAA), and cytokinins (CTKs), and transcription level of ABA biosynthesis and signaling related genes in fruit. Furthermore, a series of genes involved in biosynthesis and the metabolite pathway of sugars, anthocyanins, and fatty acids were shown to be significantly up-regulated under 500 mg/L ABA treatment. In addition, Pearson correlation analysis demonstrated that there existed relatively strong cooperativities in the ABA/kinetin (KT)-appearance parameters, ABA/IAA/KT-sugars, ABA/indolepopionic acid (IPA)/zeatin riboside (ZR)-anthocyanins, and gibberellin 3 (GA3)/methyl jasmonate (MeJA)-fatty acids, indicating that 13 kinds of endogenous phytohormones induced by ABA had different contributions to the accumulation of quality-related metabolites, while all of them were involved in regulating the overall improvement of grape fruit quality. These results laid a primary foundation for better understanding that exogenous ABA improves fruit quality by mediating the endogenous phytohormones level in grape.

Comparing Rooting Ability and Physiological Changes of Two Eucommia ulmoides Improved Varieties

Eucommia ulmoides (E. ulmoides) is a significant national strategic resource in China. It is a natural high quality rubber resource, with great development potential. We found large differences in rooting ability during adventitious root (AR) formation in two E. ulmoides improved varieties. Therefore, we used two improved varieties of E. ulmoides, ‘Huazhong 6’ (H6, with rooting rate 85.3%) and ‘Huazhong 8’ (H8, with rooting rate 22.5%) to explore the cutting rooting mechanism. In this study, we mainly determined the morphological development process of E. ulmoides cutting rooting, and compared the rooting-related indexes of the two improved varieties, and the changes in physiological indexes closely related to rooting, which include endogenous hormones, oxidases and nutrients in the phloem of the basal stem. The results showed that indole–3–acetic acid (IAA), zeatin riboside (ZR), IAA/ZR and indoleacetic acid oxidase (IAAO) were the key factors that caused big differences in rooting ability between the two E. ulmoides improved varieties. The increase in endogenous hormone IAA content and IAA/ZR value were necessary for the formation of AR. The increase in IAA content was beneficial to AR formation. The activity of IAAO was significantly negatively correlated with the rooting ability of the E. ulmoides cuttings. The high IAAO activity of the H8 cuttings led to the consumption of IAA. Although the content of IAA increased, the rooting conditions were not reached. The accumulation of nutrients before rooting also has an important effect on rooting; it is easy for cuttings to root when the carbon–nitrogen ratio (C/N) value is high. This research provides an improved understanding of the cellular and physiological underpinnings of the AR process in woody plants. In addition, it provides a theoretical basis and foundation for subsequent research on E. ulmoides cuttage technology.

Effect of Exogenous Abscisic acid (ABA) on the Morphology, Phytohormones, and Related Gene Expression of Developing Lateral Roots in ‘Qingzhen 1’

Lateral roots (LRs) are critical for plant stress tolerance and productivity. Understanding how hormones and genes interact in a fluctuating environment to coordinate LR development is a major challenge. Abscisic acid (ABA) is the primary stress-responsive hormone and mediates LR development in various plant species. However, the effect of exogenous ABA on LR development has not been elucidated in apple. In this study, ‘Qingzhen 1’ was treated with exogenous 5 µM ABA for 20 days to investigate the regulation mechanism of ABA on LR development. Morphological observations advocated that ABA inhibited both LR and shoot development in ‘Qingzhen 1’ apple plants, where the root number was 16.94%, the root length was 30.32%, the plant height was 10.88%, and the stem thickness was 8.08% lower than those in the control plants. Meanwhile, the endogenous ABA concentration was significantly increased, but the indole-3-acetic acid (IAA), zeatin riboside (ZR), and jasmonic acid (JA) concentrations were significantly decreased with ABA treatment. Furthermore, the expression levels of ABA-related genes (MdCYP707A2, MdABI1, MdAREB2, and MdABF3) were significantly upregulated, while the expression levels of auxin-related genes (MdYUCCA3, MdYUCCA8, MdPIN1 MdPIN2, MdPIN3, and MdARF19), root development-related genes (MdWOX5 and MdWOX11), and cell cycle-related genes (MdCYCD1;1 and MdCYCD3;1) were significantly downregulated at the early stage of ABA treatment, which act together on the inhibition of LR development. Taken together, the changes in hormone levels and gene expression resulted in inhibited LR development of apple plants in response to ABA.

Endogenous Hormones in Habanero Pepper Seeds

Aims: Habanero pepper (Capsicum annuum L.) cultivation is growing worldwide, mainly as a result of its high demand and nutritive contribution to human society. The presence of endogenous gibberellins and cytokinins in seeds of various vegetable crops has been related to a good germination; however, little is known on habanero pepper. The aim of this study was to search for the presence, identification and amount of gibberellins and cytokinins in seeds of habanero pepper cv “Jaguar”. Study design: Laboratory analysis for gibberellins and cytokinins were organized in solvent solutions groups with three technical replicates using a complete randomized design and results when applicable were analyzed using the statistical program 'RStudio' (version 10) and data obtained subjected to a comparison of means with the Tukey (P≤0.05) test. Place and duration of study: The experiment was conducted at the Department of Horticulture in Universidad Autónoma Agraria Antonio Narro, Saltillo, Mexico, during 2020-2021. Methodology: Lots of 50 grams dry weight of “Jaguar” habanero pepper seed samples were freeze dried and prepared with several organic solvents for the extraction, purification and identification of gibberellins and cytokinins using the techniques of GC-MS and GCMS-SIM respectively. Results: Gibberellins A1, A4, A7, A9, A15, A17 and A44 were found in habanero seed tissue. The cytokinins zeatin and zeatin-riboside were also detected in analyzed habanero samples. Conclusion: The endogenous gibberellins A1, A4, A7, A9, A15, A17, A44, and the cytokinins zeatin and zeatin-riboside are present in habanero pepper cv “Jaguar” contributing to an improve seed viability, ensuring health and overall plant yield.

Peroxidase Enzyme Fractions as Markers of Somatic Embryogenesis Capacities in Olive (Olea europaea L.)

As part of the search for biochemical markers of somatic embryogenesis in tissue cultures of olive (Olea europaea L.), peroxidases (POXs) in both the soluble and ionically wall-bound fractions were studied in two reputed olive cultivars (cvs.): “Picholine Marocaine” and “Dahbia”. In order to carry out embryogenesis induction, proximal cotyledons were cultured in modified olive medium (OMc) supplemented with 25 μM indole-3-butylic acid (IBA) and 2.5 μM 2-isopentenyladenine (2iP), while distal leaf fragments (somatic explants) were cultured in OMc supplemented with 4.56 µM zeatin riboside (ZR) and 10.25 µM 1-naphthaleneacetic acid (NAA). Regarding embryogenic potentials, the zygotic explants (cv. Picholine Marocaine: 43.39%; cv. Dahbia: 53.41%) were more regenerative than the somatic explants (cv. Picholine Marocaine: 13.05%; cv. Dahbia: 19.51%). The enzyme assay showed a higher POX activity in embryogenic calluses (ECs) than in nonembryogenic calluses (NECs) for the zygotic explants in both studied cultivars. When expressed as units per milligram of proteins (U mg−1 proteins), the highest total POXs activities (soluble POXs + ionically wall-bound POXs) were found in the ECs derived from the zygotic explants; for cv. Dahbia, 65% of the enzyme activities came from the ionically wall-bound fractions. Polyacrylamide gel electrophoresis showed that the ECs of the highly active cv. Dahbia were characterized by highly active isoperoxidases that were revealed in four migration zones, particularly a doublet in the A4 zone (Rf 0.70–0.73) present in the ionically wall-bound POXs. The fast-moving anodic POXs of the ionically wall-bound fractions could be adopted as an early electrophoretic test to determine the embryogenesis capacities in olive tissue culture materials. As biochemical markers, the POX enzyme and its profile in fractions, i.e., as soluble POXs and ionically wall-bound POXs, can offer a valuable tool for improving the tissue culture of olive via somatic embryogenesis.