As a woody plant, peony (Paeonia suffruticosa) has a long growth cycle and inefficient traditional breeding techniques. There is an urgent need in peony molecular breeding to establish an efficient and stable in vitro regeneration and genetic transformation system, in order to overcome the recalcitrant characteristics of peony regeneration and shorten the breeding cycle. The development of plant somatic embryos is an important way to establish an efficient and stable in vitro regeneration and genetic transformation system. Plant-specific WUSCHEL-related homeobox (WOX) family transcription factors play important roles in plant development, from embryogenesis to lateral organ development. Therefore, in this research, four PoWOX genes of “Fengdan” (Paeonia ostii) were cloned from the peony genome and transcriptome data of preliminary peony somatic embryos. The sequence characteristics and evolutionary relationships of the PoWOX genes were analyzed. It was demonstrated that the four PoWOX genes, named PoWOX1, PoWOX4, PoWOX11, and PoWOX13, belonged to three branches of the WOX gene family. Their expression patterns were analyzed at different stages of development and in different tissues of peony seedlings. The expression localization of the PoWOX genes was determined to be the nucleus via subcellular localization assay. Finally, the interaction protein of the PoWOX genes was identified via yeast two-hybrid assay combined with bimolecular fluorescence complementation assay. It was shown that PoWOX1 and PoWOX13 proteins could form homodimers by themselves, and PoWOX11 interacted with PoWOX1 and PoWOX13 to form heterodimers. Peony stem cell activity may be regulated from PoWOX1 and PoWOX13 by forming dimers and moving to peony stem cells through plasmodesmata. Additionally, PoWOX11–PoWOX1 and PoWOX11–PoWOX13 may play important regulatory functions in promoting the proliferation of stem cells and maintaining the homeostasis of stem cells in the SAM of peony stems. Exploring the critical genes and regulatory factors in the development of the peony somatic embryo is beneficial not only to understand the molecular and regulatory mechanisms of peony somatic embryo development but also to achieve directed breeding and improvements in efficiency through genetic engineering breeding technology to accelerate the fundamental process of molecular breeding in peony.
BABY BOOM (BBM) is one of the members of the plant-specific APETALA2/ethylene-responsive factor (AP2/ERF) transcription factor superfamily. It acts as a key regulator of plant cell pluripotency, playing a significant role in promoting somatic embryogenesis. In this study, a BBM gene named PoBBM was screened, cloned, and identified from the third-generation full-length transcriptome data of Paeonia ostii. Its open reading frame was 2136 bp, encoding 711 amino acids. Sequence feature analysis revealed that it possessed two AP2 conserved domains and eight motifs, including bbm-1. The phylogenetic tree indicated that PoBBM clusters with AtBBM in the euANT group of the Arabidopsis AP2 family, which is most closely related to grape VvBBM and may have the same ancestry as grape. Subcellular localization demonstrated that the PoBBM protein was localized in the nucleus. Semi-quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to assess the PoBBM transcript levels during ten developmental stages of somatic embryos and in five tissue types of peonies. The results indicate that PoBBM was highly expressed in the early stages of peony somatic embryo development. The expression on 0–15 d was the highest and decreased gradually with somatic embryogenesis. The gene is almost not expressed after 40 d since somatic embryo formation. PoBBM was expressed in roots, stems, leaves, seeds, and calli, with the highest levels in seeds, followed by leaves and calli. The PoBBM protein displayed transcriptional self-activation activity, which may facilitate further research on its relationships with other proteins. The above results provide a key gene PoBBM for somatic embryogenesis in peonies, which is significant for advancing the establishment of a stable and efficient regeneration and genetic transformation system for peonies.
Moso bamboo (Phyllostachys edulis (Carriere) J. Houzeau), native to China, is one of the most economically and ecologically important bamboo species. Since the economic interests and the strong clonality, it has been widely cultivated in southern China, which inevitably reduces the natural stands and leads to gene loss in this species. In this study, three natural populations of Moso bamboo distributed in Anhui, Guangxi, and Zhejiang province, were used to analyze the correlation between phenotypic traits, cell structure, and material properties from the perspective of phenotypic, genetic, and environmental. Among those traits and properties, fiber width was correlated with wall thickness at breast height and average nodes length under branch positively. Leaf length was correlated positively with fiber lumen diameter and parenchyma lumen diameter. Furthermore, it showed a very close correlation between moisture content, bending strength, modulus of elasticity, and diameter at breast height, clear height, and leaf length. The lumen diameter of fiber cell wall thickness is positively correlated with bending strength and modulus of elasticity. Density is positively correlated with parenchyma cell wall thickness. The experimental design is relatively detailed and representative, and the workload is huge. This study reflects the research objectives with scientific and rational experiments and data. This study will analyze the differences of various indicators from the perspective of genetic to build a bridge between micro-structure and macro-structure for rational utilization of the whole area of Moso bamboo resources in China.
Plant-specific WUSCHEL (WUS)-related homeobox (WOX) family of transcription factors are involved in apical meristem maintenance, embryogenesis, lateral organ development, and hormone signaling. Among the members of this family, WOX1 is known to play essential roles in many species. However, the function of the peony ‘Feng Dan’ (Paeonia ostii L.) WOX1 (PoWOX1) remains unknown. The initial bioinformatic analysis revealed that PoWOX1 belongs to the modern clade of the WOX gene family and has a highly conserved homeodomain (HD), the WUS motif, the STF-box, and the MAEWEST/WOX4-box. Subsequent heterologous overexpression in Arabidopsis thaliana revealed that PoWOX1 promotes root growth, early shoot initiation, and flowering. The root vascular tissues, especially the arrangement and size of xylem cells, were different between the PoWOX1-overexpressing transgenics and the wild-type plants, and the pericycle cells adjacent to the xylem divided more easily in the transgenics than in the wild type. Furthermore, under in vitro conditions, the transgenic leaf explants exhibited more callus induction and differentiation than the wild-type leaf explants. Thus, the study’s findings provide novel insights into the role of PoWOX1 in promoting root development and callus tissue induction and differentiation, serving as a reference for developing an efficient regeneration system for the peony.
In recent years, bamboo has been well exploited in the pulp and paper industry. Moso bamboo (Phyllostachys edulis (Carriere) J. Houzeau; hereafter M), originated from China with a long history of cultivation as the most abundant resource with the widest distribution area among bamboo plants in China. In this study, Moso bamboo (M) and nine bamboo varieties were selected in the bamboo germplasm resources storage bank of the Anhui Taiping Experimental Station of ICBR. The characteristics of their cell structures were compared, and the differences were analyzed from a genetic perspective. The results showed that M had the highest fiber cell length, fiber cell width, length, and parenchyma width, while GJ showed the lowest of these measurements [P. edulis ’Kikko-chiku’,G.H.Lai]. The fiber wall thickness of Q [P. edulis f. obliquinoda (Z.P.Wang et N.X.Ma) Ohrnberger] was the smallest, while its fiber lumen diameter was the highest in the group. The parenchyma wall thickness and parenchyma lumen diameter of Q were the smallest in the group. The fiber cells of M and Q had better flexibility, which is conducive to improving the tensile strength, break resistance, and folding resistance of paper made from these materials. SY and GJ may be more suitable for ornamental items because of their special appearances. The purpose of this study was to explore the genetic variation patterns of various cell structure indicators among Moso bamboo and its varieties, as well as to develop a strategy of bamboo growing and lumbering based on the local conditions, providing reference data for the utilization of non-woody forest resources.
AINTEGUMENTA-LIKE6 (AIL6) is a transcription factor specifically expressed in embryos. It is a key to improving peony regeneration through tissue culture. Based on transcriptomic data from our previous research and the published genome data of Paeonia ostii, we identified the PoAIL6 gene associated with somatic embryogenesis (SE) in Paeonia ostii ‘Fengdan’. Structural and phylogenetic analyses were conducted on the PoAIL6-encoded protein, and its expression pattern across tissues and embryo developmental stages were explored using real-time quantitative PCR. Our results revealed that PoAIL6 contained two AP2 conserved domains and the characteristic motif of AIL6 genes. Phylogenetic analysis revealed that the PoAIL6 gene has similarity to grape (Vitis vinifera) and cocoa (Theobroma cacao). PoAIL6 exhibited the highest expression during early embryonic development, with expression levels gradually decreasing throughout SE progression. It was most highly expressed in peony seeds and showed relatively high expression in callus tissue. This study underscores the pivotal role of PoAIL6 in the early SE and lays a playground for elucidating its molecular mechanisms, supporting the development of efficient and stable regeneration and transformation systems in peony.