Bud Development Research Articles

Comprehensive genome-wide characterization of the MIKC-type MADS-box family members and the dynamic expression profiling throughout the development of floral buds in Populus tomentosa

Poplar is a major silvicultural tree species for industrial production, with versatile applications in construction, furniture, pulp and biofuel. The phenotypic appearance of male or female floral buds in poplar trees, exhibits a strikingly similar morphology at corresponding developmental stages. However, there are significant differences in internal anatomical structures. The MIKC-type MADS-box transcription factor family plays an indispensable role in regulating the development of floral organs and increasing vegetative biomass, and it is also of great significance in elucidation of the aforementioned morphological differences. This study systematically analyzed the MIKC-type MADS-box transcription factor family in Populus tomentosa. A total of 100 MIKC-type MADS-box genes were identified, which were divided into 14 subfamilies. We examined the physicochemical properties, gene structure, conserved motifs, chromosome distribution, collinearity, promoter cis-acting elements, gene expression profiles, and protein-protein interaction network of these 100 MIKC-type MADS-box members. Excitingly, four clusters of PtMADS members exhibited a high level of abundant expression in the initial and mature floral buds of both male and female, suggesting critical significance in tuning the morphogenesis and development of floral organs, as well as in the modulation of reproductive fitness. The protein-protein interaction network diagram corroborated these findings, substantiating the speculated roles of these genes. Our findings lay a foundational framework for future functional explorations and potential applications of MIKC-type MADS-box genes in this industry tree species.

Plant hormone signaling is involved in regulating flower bud size of daylily

To explore the role and metabolic differences of plant hormones in regulating the flower bud size of daylily, we collected the flower buds from two daylily varieties 'Datong Huanghua' and 'Dongbei Huanghua' at the young bud, green, yellowing, and yellow stages for transcriptome sequencing. The differentially expressed genes (DEGs) were screened, and the exogenous plant hormone spraying experiments were conducted. A total of 199 DEGs related to the biosynthesis and metabolism of plant hormones was screened out at different flower development stages of 'Datong Huanghua' and 'Dongbei Huanghua'. These genes regulated the biosynthesis and metabolism of six plant hormones: abscisic acid, gibberellin, auxin, jasmonate, cytokinin, and ethylene. The DEGs associated with auxin were the most, which suggested that auxin played a role in regulating flower bud development. The auxin response factor (ARF) presented up-regulated expression at all the four stages of flower bud development, indicating that ARF played a positive regulatory role throughout the flower bud development of daylily. The experiments with exogenous spraying of six hormones further verified that indole-3-acetic acid (IAA) significantly promoted the growth and increased the nutrient content in the flower buds of 'Datong Huanghua', suggesting that IAA played a role in regulating flower bud development. Our results laid a theoretical foundation for probing into the regulatory mechanism of flower bud development of 'Datong Huanghua' and 'Dongbei Huanghua'.

Bioenergy sorghum nodal root bud development: morphometric, transcriptomic and gene regulatory network analysis.

Bioenergy sorghum's large and deep nodal root system and associated microbiome enables uptake of water and nutrients from and deposition of soil organic carbon into soil profiles, key contributors to the crop's resilience and sustainability. The goal of this study was to increase our understanding of bioenergy sorghum nodal root bud development. Sorghum nodal root bud initiation was first observed on the stem node of the 7th phytomer below the shoot apex. Buds were initiated near the upper end of the stem node pulvinus on the side of the stem opposite the tiller bud, then additional buds were added over the next 6-8 days forming a ring of 10-15 nascent nodal root buds around the stem. Later in plant development, a second ring of nodal root buds began forming on the 17th stem node immediately above the first ring of buds. Overall, nodal root bud development can take ~40 days from initiation to onset of nodal root outgrowth. Nodal root buds were initiated in close association with vascular bundles in the rind of the pulvinus. Stem tissue forming nascent nodal root buds expressed sorghum homologs of genes associated with root initiation (WOX4), auxin transport (LAX2, PIN4), meristem activation (NGAL2), and genes involved in cell proliferation. Expression of WOX11 and WOX5, genes involved in root stem niche formation, increased early in nodal root bud development followed by genes encoding PLTs, LBDs (LBD29), LRP1, SMB, RGF1 and root cap LEAs later in development. A nodal root bud gene regulatory network module expressed during nodal root bud initiation predicted connections linking PFA5, SPL9 and WOX4 to genes involved in hormone signaling, meristem activation, and cell proliferation. A network module expressed later in development predicted connections among SOMBRERO, a gene involved in root cap formation, and GATA19, BBM, LBD29 and RITF1/RGF1 signaling. Overall, this study provides a detailed description of bioenergy sorghum nodal root bud development and transcriptome information useful for understanding the regulation of sorghum nodal root bud formation and development.

Effect of Organic Substrates on Macro-propagation of Banana under Island Conditions

Banana is an important tropical fruit crop grown globally and is the world’s most important agricultural food commodity. Plantation crops take a major share of cultivated area in Andaman and Nicobar Islands. Banana is one of the important fruit crops that is highly suitable to grow as intercrop in plantation based cropping system. Local varieties of banana like Korangi, Mitta Champa, Khatta Champa and Cheenakela are cultivated by the farmers traditionally to meet the local demand. However, in recent past growing commercial varieties of banana has become popular in the Island due to consumer demand. Traditionally banana is propagated by suckers and wide variability was observed in sucker production among the commercial varieties. Due to this large scale multiplication of banana through suckers could not meet the planting material demand in the Island. Hence, a study was undertaken during 2020-2022 to standardize the macro-propagation technology using different organic substrates in the commercial variety ‘Poovan’. Nine different organic substrate combinations were used in the present study to enhance the efficacy of lateral bud development and plantlet production. Suckers weighing 1.0 to 1.5 kg were used as the propagule. Among all the organic media combinations tested, significant differences were observed in terms of plantlet production, root and shoot growth. The treatment T8 (Coircompost: sawdust: FYM: vermicompost (1:1:1:1) with Arka microbial consortia @10 kg) produced maximum number of primary (5.60) and secondary buds (12.07) during primary and secondary capitation of decorticated mother rhizome. Early bud emergence (11.5 days), maximum number of plantlets (12-13 per mother rhizome), maximum shoot length (46.43 cm), maximum shoot girth (3.03 cm) and maximum number of leaves (4.50) were also observed in the treatment T 8. Minimum number of days was taken (52.70 days) for plantlet separation during secondary capitation in the same treatment T 8 suggesting that the treatment combination (Coircompost: sawdust: FYM: vermicompost (1:1:1:1) with Arka microbial consortia @10 kg) was effective for macro-propagation of banana Cv. Poovan under Island ecosystem.

Development of Ectodermal and Endodermal Taste Buds.

The sense of taste is mediated primarily by taste buds on the tongue. These multicellular sensory organs are induced, patterned and become innervated during embryogenesis such that a functional taste system is present at birth when animals begin to feed. While taste buds have been considered ectodermal appendages, this is only partly accurate as only fungiform taste buds in the anterior tongue arise from the ectoderm. Taste buds found in the posterior tongue actually derive from endoderm. Nonetheless, both anterior and posterior buds are functionally similar, despite their disparate embryonic origins. In this review, I compare the development of ectodermal vs endodermal taste buds, highlighting the many differences in the cellular and molecular genetic mechanisms governing their formation.

Enhancement of bud dormancy release and development, leaf nutrition, flower and fruit quality of kiwifruit cv. ‘Hayward’ induced by BUD 14 biostimulant

The aim of this research was to investigate the influence of a foliar fertilization program, consisted of the BUD 14 nitrogen-calcium commercial formulation (N: 14% w/w, CaO: 5.5% w/w) as a biostimulant, on bud development percentage, flowering rate, classification of flowers into open, closed and triple, flower and pollen quality traits, fruit quality attributes, and leaf nutritional status of the ‘Hayward’ kiwifruit cultivar. The study was realized during a two-year experimental period in Naoussa, Central Macedonia, Greece. The results showed that BUD 14 induced synchronization in bud development relative to different vegetative stages including initiation of bud expansion, appearance of leaf apices covered by hair and deployment of 2-8 leaves and increased the flowering rate of open flowers. Pedicel length, ovary fresh weight, and dry weight, dry matter and length in female flowers as well as maximum pollen grain diameter and area in polar view in male flowers were significantly enhanced in the BUD 14 treatment. Fruit quality characteristics like average weight and dry mass were significantly augmented, and a 1.5-fold and 2-fold increase was recorded in canes length and number of kiwifruits per cane. In addition, leaf nutrient Ca and Mg concentrations were significantly enhanced, compared to the control. The efficacy of BUD 14 as a more target-oriented and environmentally friendly alternative method of supplying plants with smaller and controlled amounts of nutrients for breaking bud dormancy and improving their development was demonstrated, enhancing flower and fruit quality, leaf nutrition, kiwifruit developmental characteristics, and finally the total production per fruit per tree.

CpSPL10-CpELF4 module involves in the negative regulation of flower bud differentiation in Chinese cherry

SQUAMOSA promoter-binding protein-like (SPL) genes play a crucial role in regulating floral induction. Despite such importance, a comprehensive study of SPLs in Chinese cherry flower bud development has been absent. In this study, 32 CpSPL genes were identified. According to expression profiling, CpSPLs exhibited tissue-specific expression and distinct trends throughout flower bud differentiation. Specifically, CpSPL10 was greatly expressed at the beginning of the differentiation, and its role was further investigated. Its overexpression extended the vegetative growth of transgenic tobacco plants, delayed flowering by about 20 days. Moreover, the accumulation of NbELF4 (Early flowering 4) transcripts was enhanced due to the up-regulated levels of CpSPL10 in tobacco plants. ELF4 functions as a major element of the circadian clock; its high expression typically delays the transition from vegetative-to-reproductive growth. Further experiments revealed that CpSPL10 interacts with CpSPL9 or a transposase-derived transcription factor CpFRS5 (FAR1-RELATED SEQUENCE 5) and activates the expression of the downstream gene CpELF4. Notably, the GUS fusing reporter assay detected the activation of CpSPL10 and CpELF4 promoters in shoot apical meristems of transgenic Arabidopsis. These findings revealed the negative regulation of the CpSPL10-CpELF4 module in flower bud differentiation, providing references for supplementing the specific relationships among SPL, FRS, and ELF4.

RhLHY-RhTPPF-Tre6P Inhibits Flowering in Rosa hybrida Under Insufficient Light by Regulating Sugar Distribution.

Light is crucial for flower bud development in plants, serving as both signal and energy source. However, the mechanisms by which daylength and light intensity regulate flowering in modern roses remain unclear. In Rosa hybrida 'Carola', insufficient light delays flowering and reduces the sugar content in terminal buds. RNA sequencing identified the Trehalose-6-phosphate phosphatase F (RhTPPF) gene as a key responder to insufficient light, modulating Tre6P metabolism. Overexpression of RhTPPF in rose calli enhanced sugar accumulation and suppressed the synthesis of RhCO/FT. In tobacco, overexpression of RhTPPF delayed the transition from vegetative growth to flowering, while silencing RhTPPF in roses accelerated flowering. Silencing RhTPPF in roses elevated trehalose-6-phosphate (Tre6P) levels and decreased trehalose. Transcriptome data showed that the expression level of RhTPPF was highly correlated with the circadian rhythm gene LATE ELONGATED HYPOCOTYL (RhLHY). Yeast one-hybrid assays, dual luciferase assays and EMSA revealed that RhLHY directly binds to the RhTPPF promoters. Overexpression of RhLHY suppressed flowering, while silencing RhLHY promoted flowering. Furthermore, altering the expression of RhLHY influenced Tre6P synthesis and the expression of sucrose-related transport genes. These findings suggest a RhLHY-RhTPPF-Tre6P regulatory module that maintains sugar balance and inhibits flower formation under reduced light conditions by modulating sugar distribution.

In Vitro Micropropagation of Eria Lasiopetala (Willd.) Ormerod- A Native Medicinal Orchid in Bangladesh

The present study focuses on the asymbiotic germination of seeds through in vitro culture of Eria lasiopetala, a native medicinal orchid in Bangladesh. Out of the four basal media, MS was found to be the most effective medium, where 93.58±0.374% seed germinations were recorded. Different plant growth regulators (PGRs), e.g. BAP, Kin, NAA and were used for secondary protocorm proliferation, plantlet development, and multiplication of shoot buds through in vitro conditions. The maximum number of secondary protocorms (12.80±0.115) were developed that were derived from primary protocorms on MS medium fortified with 0.5 mg/l BAP and 1.0 mg/l NAA. Data revealed that 0.5 mg/l BAP + 0.25 mg/l NAA showed the best combination for height elongation of plantlets (3.12±0.012 cm) in MS medium. While for multiple shoot buds (MSBs) 1.0 mg/l BAP + 0.5 mg/l NAA showed better (6.8±0.127) performance than other combinations of PGRs. The single shoots were isolated from the multiple shoots and sub-cultured on ½MS medium supplemented with NAA (0.5 - 2.0 mg/l), IAA (0.5 - 2.0 mg/l) and IBA (0.5 - 2.0 mg/l) for root induction. The maximum number of roots (7.53±0.176) and its growth (4.61±0.01 cm) were found in ½MS medium supplemented with 1.0 mg/l IAA. The well rooted plantlets were hardened and successfully transferred after acclimatization in pots containing the mixture of coconut husk, charcoal and brick pieces in the ratio of 2:1:1 and eventually established under natural condition. J. Bio-Sci. 32(1): 45-60, 2024

Integrated metabolomic and transcriptomic analysis revealed the transition of functional components in edible flower buds of Hemerocallis citrina Baroni

The edible flower buds of Hemerocallis citrina Baroni are used both as a vegetable and functional food. It has various health benefits due to the diversity of natural products. However, the establishment of functional components in the edible flower bud remains to be studied. We conducted a high-resolution metabolomic analysis of flower buds at three developmental stages, 1–2 cm, 4–6 cm, and edible (10–15 cm). Our analysis revealed 157 differential accumulated metabolites, including flavonoids (49), fatty acids (17) and terpenoids (13) while most of them decreased during flower bud development. Among them, 2 flavonoids, 2 long-chain fatty acids and 1 triterpene saponin are highly accumulated in edible flower buds. Furthermore, the expression levels of catalytic genes mirrored the changes in metabolite levels detected. These results track the dynamics of functional component accumulation during edible flower bud development, laying the theoretical basis for nutrition formation in H. citrina.

Optimizing Fertilization Strategies to Promote Leaf-Use Ginkgo Productivity and Ecosystem Economic Benefits: An Integrated Evaluation of a Field Trial in Southern China

Field experiments were conducted on a four-year-old leaf-use ginkgo plantation in southern China to assess the impact of nine different fertilization strategies with varying N-P2O5-K2O rates at three growth phases (FBD: March for bud development; FLG: May for leaf growth; FLS: July for leaf strengthening) on leaf-use ginkgo (Ginkgo biloba L.) leaf productivity and ecological economic benefits (EEBs). The results indicated that regardless of timing and rate, fertilizer application led to an increase in leaf area and thickness, resulting in higher ginkgo leaf yield. The highest fresh (215.14 g tree−1) and dry (78.83 g tree−1) yields were observed with 3 g N + 2.5 g P2O5 + 1.5 g K2O tree−1 in FLG. FLS was found to mitigate the decline in SPAD values of leaves during late summer. Furthermore, fertilized ginkgo trees exhibited higher flavonoid concentrations in leaves, enhancing profitability. However, higher fertilizer rates were associated with elevated greenhouse gas emissions, nitrogen losses and ecological costs. Despite these drawbacks, all fertilization treatments resulted in increased net economic income. Specifically, compared to no fertilization, FBD, FLG and FLS treatments boosted net income by 3.5~26.6%, 11.6~60.5% and 5.8~35.4%, respectively. Using the entropy weight TOPSIS method, it was concluded that optimizing the N, P and K fertilization rate and timing (applying 3–2.5–1.5 g tree−1 of N-P2O5-K2O in May) is a beneficial approach to maximize EEBs and industrial benefits in leaf-use ginkgo plantations in southern China. This study provides valuable insights into suitable fertilization patterns and management for leaf-use ginkgo plantations in southern China.

Pollen viability, stigma receptivity and reproduction mode of spineless cactus (Nopalea cochenillifera Salm-Dyck) accessions

Forage cactus (Opuntia spp. and Nopalea spp.) have great potential for animal feed and human consumption due to their adaptability to semi-arid conditions. This study evaluated six accessions of N. cochenillifera, analyzing pollen viability, stigma receptivity and reproduction mode. Four types of pollination were tested: spontaneous self-pollination, manual self-pollination, natural pollination and cross-pollination. The results indicated high pollen viability and stigma receptivity in some accessions during flower bud development, allowing for self-pollination and controlled hybridization. After 24 hours of anthesis, pollen viability decreased, but stigma receptivity reached 100%. Fruiting occurred via natural pollination in all accessions, and by self-pollination and cross-pollination in accessions ac-02 and ac-03, which showed a mixed reproductive system. These results are essential for breeding programs, contributing to sustainable agriculture in semi-arid regions.

Possible Mechanism of Sucrose and Trehalose-6-Phosphate in Regulating the Secondary Flower on the Strong Upright Spring Shoots of Blueberry Planted in Greenhouse.

Secondary flowering is the phenomenon in which a tree blooms twice or more times a year. Along with the development of blueberry (Vaccinium corymbosum L.) fruits in spring, a large number of secondary flowers on the strong upright spring shoots were noticed in blueberries planted in the greenhouse. To reveal the cause and possible regulatory mechanism of the phenomenon, we clarified the phenological characteristics of flower bud differentiation and development on the spring shoots by combining phenological phenotype with anatomical observation. Furthermore, the changes in carbohydrates, trehalose-6-phosphate (Tre6P), and the relationship among the key enzyme regulatory genes for Tre6P metabolism and the key regulatory genes for flower formation during the differentiation process of apical buds and axillary buds were investigated. The results showed that the process of flower bud differentiation and flowering of apical and axillary buds was consistent, accompanied by a large amount of carbohydrate consumption. This process was positively correlated with the expression trends of VcTPS1/2, VcSnRK1, VcFT, VcLFY2, VcSPL43, VcAP1, and VcDAM in general, and negatively correlated with that of VcTPP. In addition, there is a certain difference in the differentiation progress of flower buds between the apical and axillary buds. Compared with axillary buds, apical buds had higher contents of sucrose, fructose, glucose, Tre6P, and higher expression levels of VcTPS2, VcFT, VcSPL43, and VcAP1. Moreover, VcTPS1 and VcTPS2 were more closely related to the physiological substances (sucrose and Tre6P) in axillary bud and apical bud differentiation, respectively. It was suggested that sucrose and trehalose-6-phosphate play a crucial role in promoting flower bud differentiation in strong upright spring shoots, and VcTPS1 and VcTPS2 might play a central role in these activities. Our study provided substantial sight for further study on the mechanism of multiple flowering of blueberries and laid a foundation for the regulation and utilization of the phenomenon of multiple flowering in a growing season of perennial woody plants.

Coding and description of phenological stages of blue honeysuckle (Lonicera caerulea L.) according to the BBCH scale

Blue honeysuckle (Lonicera caerulea L.) is an emerging fruit crop from the temperate to circumpolar climate regions with outstanding agronomic traits, substantial nutritional value, and versatile processing applications. However, the limited understanding of its phenology poses substantial challenges to its cultivation, breeding, and research. In this study, we characterized the progressive phenological growth stages of blue honeysuckle according to the BBCH (Biologische Bundesanstalt, Bundessortenamt und Chemische Industrie) scale by investigating three major cultivars (L. caerulea) and a wild accession (L. caerulea var. edulis) in Harbin, China (hot-summer humid continental climate) during 2022–2023. The annual growth of blue honeysuckle was characterized by eight primary stages, including bud development (0), leaf development (1), shoot development (3), inflorescence emergence (5), flowering (6), fruit development (7), fruit ripening (8), and senescence and dormancy (9). Additionally, 35 secondary growth stages were documented, among which eight stages are related to fruit development and ripening. The annual growth period of blue honeysuckle spans 6.5–7 months, starting from late March and ending in early October. The flowering season lasts for 2–3 weeks, ranging from late April to mid-May. The fruit development and ripening last for 7–9 weeks, beginning in early May and ending in early July. This study provides valuable insights for the cultivation and research of blue honeysuckle.

Transcriptomic and Physiological Analyses for the Role of Hormones and Sugar in Axillary Bud Development of Wild Strawberry Stolon.

Strawberries are mainly propagated by stolons, which can be divided into monopodial and sympodial types. Monopodial stolons consistently produce ramets at each node following the initial single dormant bud, whereas sympodial stolons develop a dormant bud before each ramet. Sympodial stolon encompasses both dormant buds and ramet buds, making it suitable for studying the formation mechanism of different stolon types. In this study, we utilized sympodial stolons from Fragaria nilgerrensis as materials and explored the mechanisms underlying sympodial stolon development through transcriptomic and phytohormonal analyses. The transcriptome results unveiled that auxin, cytokinin, and sugars likely act as main regulators. Endogenous hormone analysis revealed that the inactivation of auxin could influence bud dormancy. Exogenous cytokinin application primarily induced dormant buds to develop into secondary stolons, with the proportion of ramet formation being very low, less than 10%. Furthermore, weighted gene co-expression network analysis identified key genes involved in ramet formation, including auxin transport and response genes, the cytokinin activation gene LOG1, and glucose transport genes SWEET1 and SFP2. Consistently, in vitro cultivation experiments confirmed that glucose enhances the transition of dormant buds into ramets within two days. Collectively, cytokinin and glucose act as dormant breakers, with cytokinin mainly driving secondary stolon formation and glucose promoting ramet generation. This study improved our understanding of stolon patterning and bud development in the sympodial stolon of strawberries.

Micropropagation and genetic transformation of Byblis liniflora

AbstractByblis, a small genus of carnivorous plants predominantly found in Australia, is characterized by its passive trapping mechanism and unique floral features. The chemical composition of Byblis, including identified phenylethanoid glycosides, particularly acteoside, highlights its pharmacological potential with various biological activities. In vitro culture techniques have been established for propagation, with micropropagation protocols developed for different Byblis species. However, information on genetic transformation, vital for trait modification and enhanced pharmacological interest, remains limited. This study focuses on optimizing micropropagation, adventitious regeneration, and genetic transformation methods for Byblis liniflora. Adventitious regeneration rates were highest in medium with reduced Murashige and Skoog salts (MS/10) and sucrose (3 gL−1) concentrations. Zeatin supplementation (1 mgL−1) further improved regeneration rates and bud development with 100% of regenerated root explants and 8.8 shoots per explant. Liquid MB3 medium supplemented with indole-3-acetic acid (IAA) 5 mgL−1 facilitated efficient rooting and acclimatization. The establishment of an efficient Rhizobium-mediated genetic transformation method yielded transgenic plants expressing green fluorescent protein (GFP). Molecular analysis confirmed transgene integration, marking the first successful genetic transformation in the Byblis genus. These advancements pave the way for exploring gene function and enhancing pharmacological properties, thereby broadening our understanding and utilization of carnivorous plants like Byblis.

Morphological and cytological assessments reveal pollen degradation causes pollen abortion in cotton cytoplasmic male sterility lines

BackgroundUnderstanding the mechanism of male sterility is crucial for producing hybrid seeds and developing sterile germplasm resources. However, only a few cytoplasmic male sterility (CMS) lines of cotton have been produced due to several challenges, like inadequate variation of agronomic traits, incomplete sterility, weak resilience of restorer lines, and difficulty in combining strong dominance. Therefore, the morphological and cytological identification of CMS in cotton will facilitate hybrid breeding.ResultsTwo F2 segregating populations of cotton were constructed from cytoplasmic male sterile lines (HaA and 01A, maternal) and restorer lines (HaR and 26R, paternal). Genetic analysis of these populations revealed a segregation ratio of 3:1 for fertile to sterile plants. Phenotypic analysis indicated no significant differences in traits of flower bud development between sterile and fertile plants. However, sterile plants exhibited smaller floral organs, shortened filament lengths, and anther atrophy on the flowering day in comparison with the fertile plants. When performed scanning electron microscopy (SEM), the two F2 populations revealed morphological variations in the anther epidermis. Cellular analysis showed no significant differences in pollen development before pollen maturation. Interestingly, between the pollen maturation and flowering stages, the tapetum layer of sterile plants degenerated prematurely, resulting in abnormal pollen grains and gradual pollen degradation.ConclusionThe results of this study suggest that fertility-restoring genes are controlled by a single dominant gene. Sterile plants exhibit distinctive floral morphology, which is characterized by stamen atrophy and abnormal anthers. Pollen abortion occurs between pollen maturity and flowering, indicating that premature tapetum degradation may be the primary cause of pollen abortion. Overall, our study provides a theoretical basis for utilizing CMS in hybrid breeding and in-depth investigation of the dominant configuration of cotton hybrid combinations, mechanisms of sterility, and the role of sterile and restorer genes.

Sall4 regulates downstream patterning genes during limb regeneration

Many salamanders can completely regenerate a fully functional limb. Limb regeneration is a carefully coordinated process involving several defined stages. One key event during the regeneration process is the patterning of the blastema to inform cells of what they must differentiate into. Although it is known that many genes involved in the initial development of the limb are re-used during regeneration, the exact molecular circuitry involved in this process is not fully understood. Several large-scale transcriptional profiling studies of axolotl limb regeneration have identified many transcription factors that are up-regulated after limb amputation. Sall4 is a transcription factor that has been identified to play essential roles in maintaining cells in an undifferentiated state during development and also plays a unique role in limb development. Inactivation of Sall4 during limb bud development results in defects in anterior-posterior patterning of the limb. Sall4 has been found to be up-regulated during limb regeneration in both Xenopus and salamanders, but to date it function has been untested. We confirmed that Sall4 is up-regulated during limb regeneration in the axolotl using qRT-PCR and identified that it is present in the skin cells and also in cells within the blastema. Using CRISPR technology we microinjected gRNAs specific for Sall4 complexed with cas9 protein into the blastema to specifically knockout Sall4 in blastema cells only. This resulted in limb regenerate defects, including missing digits, fusion of digit elements, and defects in the radius and ulna. This suggests that during regeneration Sall4 may play a similar role in regulating the specification of anterior-proximal skeletal elements.

The tomato WRKY-B transcription factor modulates lateral branching by targeting BLIND, PIN4, and IAA15.

Lateral branching is a crucial agronomic trait that impacts crop yield. In tomato ( Solanum lycopersicum ), excessive lateral branching is unfavorable and results in substantial labor and management costs. Therefore, optimizing lateral branching is a primary objective in tomato breeding. Although many genes related to lateral branching have been reported in tomato, the molecular mechanism underlying their network remains elusive. In this study, we found that the expression profile of a WRKY gene, WRKY-B (for WRKY-BRANCING), was associated with the auxin-dependent axillary bud development process. Wrky-b mutants generated by the CRISPR/Cas9 editing system presented fewer lateral branches, while WRKY-B overexpression lines presented more lateral branches than did wild-type plants. Furthermore, WRKY-B can directly target the well-known branching gene BLIND (BL) and the auxin efflux carrier gene PIN4 to activate their expression. Both the bl and pin4 mutants exhibited reduced lateral branching, similar to the wrky-b mutant. The IAA contents in the axillary buds of the wrky-b, bl, and pin4 mutant plants were significantly higher than those in the wild-type plants. In addition, WRKY-B can also directly target the AUX/IAA gene IAA15 and repress its expression. In summary, WRKY-B works upstream of BL, PIN4, and IAA15 to regulate the development of lateral branches in tomato.

Identification of Apple Flower Development-Related Gene Families and Analysis of Transcriptional Regulation.

Apple (Malus domestica Borkh.) stands out as a globally significant fruit tree with considerable economic importance. Nonetheless, the orchard production of 'Fuji' apples faces significant challenges, including delayed flowering in young trees and inconsistent annual yields in mature trees, ultimately resulting in suboptimal fruit yield due to insufficient flower bud formation. Flower development represents a pivotal process influencing plant adaptation to environmental conditions and is a crucial determinant of successful plant reproduction. The three gene or transcription factor (TF) families, C2H2, DELLA, and FKF1, have emerged as key regulators in plant flowering regulation; however, understanding their roles during apple flowering remains limited. Consequently, this study identified 24 MdC2H2, 6 MdDELLA, and 6 MdFKF1 genes in the apple genome with high confidence. Through phylogenetic analyses, the genes within each family were categorized into three distinct subgroups, with all facets of protein physicochemical properties and conserved motifs contingent upon subgroup classification. Repetitive events between these three gene families within the apple genome were elucidated via collinearity analysis. qRT-PCR analysis was conducted and revealed significant expression differences among MdC2H2-18, MdDELLA1, and MdFKF1-4 during apple bud development. Furthermore, yeast two-hybrid analysis unveiled an interaction between MdC2H2-18 and MdDELLA1. The genome-wide identification of the C2H2, DELLA, and FKF1 gene families in apples has shed light on the molecular mechanisms underlying apple flower bud development.